Diabetes is a metabolic complication distinguished mainly by persistent hyperglycemia and is involved in the formation of reactive oxygen species, thereby causing oxidative stress, which is the major culprit for diabetic complications in different organs, including the vascular system. Controlling blood glucose is the most effective strategy for preventing diabetes and its complications. Currently, the available antidiabetic therapy is associated with several side effects, thus inexhaustible attention has been paid toward the development of natural compounds. The present review highlights the different types of flavonoids as potent antidiabetic agents along with their structure–activity relationship (SAR) studies, which will definitely aid in designing innovative molecules with improved antidiabetic efficacy. The type of substitution in the flavonoid core structure decides their bindings at different biological targets involved in diabetes development such as xanthine oxidase inhibitors, SGLT-II inhibitors, α-glucosidase inhibitors, PPAR-γ agonists, DPP-4 inhibitors, and glycogen phosphorylase inhibitors. Based on SAR investigation, a double bond between C₂ and C₃ positions, hydroxy substitutions at C₅ and C₇ positions of ring A, and substitution by the ketonic group at the C₄ position were considered as lead modifications in the bioactivity of flavonoids for potent antidiabetic activity.

Background: Baicalin, a flavone glucuronide exhibiting profuse pharmacological activities present in Oroxylum indicum stem barks. Our research highlights the process parameter at which a high yield of baicalin can be extracted from O. indicum stem barks in “one run.” Objectives: The study compares the efficacy of different modern and traditional methods for baicalin extraction. Box–Behnken design (BBD) was availed for optimization of process parameters for the extraction of baicalin from O. indicum stem barks. Materials and Methods: Extraction conditions (extraction time, solvent-to-drug ratio, and extraction temperature) were optimized by response surface methodology (RSM), specifically BBD. Quantification analysis of baicalin in different extracts was done using high-performance liquid chromatography. Results: Ultrasound-assisted extraction (UAE) method conferred the highest yield of baicalin, and ethanol was found to be the most efficient extractive solvent. Through the use of BBD, the optimal conditions for baicalin extraction were established as extraction time—29.058 min, solvent-to-drug ratio—21.124 ml/g, and extraction temperature—67.963°C. Under such conditions, baicalin was yielded as 26.572 mg/g, which was nearly close to the predicted value of 27.16 mg/g. Conclusion: The UAE method stood out to be the best among all the other thermal and non-thermal modes of extraction used, and ethanol proves to be the most efficient extracting solvent. Additionally, baicalin extraction was significantly affected by all three different variables. Our study highlights the use of RSM, a modern-day statistical technique in the extraction field of therapeutically potent phytocompounds, which makes the optimization method cheap and less laborious than the traditional optimization method.

Background: Complex Great Powerful Blend (GPB) (a great powerful blend) is a hot water extract of three well-known dried fruits: goji berry, mulberry, and jujube. These fruits have been used as food for a long time, and this article sought to confirm the immunostimulatory effects of Complex GPB. Materials and Methods: To do this, in vitro study using RAW264.7 cells and in vivo using normal mice were performed. Cytokine production and NK cell activity levels were measured by enzyme-linked immunosorbent assay, real-time polymerase chain reaction (PCR), and the YAC-1 cell-killing assay. Transcription factor activation levels were also determined by western blotting analysis. Results: First, it was confirmed that Complex GPB had no cytotoxic effects on the cells and increased the production levels of tumor necrosis factor (TNF)-a in a concentration-dependent manner. Additionally, Complex GPB enhanced pro-inflammatory cytokine expressions such as TNF-a, IL-12a, and IL-1b. Western blotting analysis revealed that activation of the NF-kB and AP-1 pathways are involved. Furthermore, in vivo study confirmed that TNF-a was found to be elevated in the serum and splenocytes of mice-fed Complex GPB. Cytotoxic activation of NK cells was also increased in mice injected with Complex GPB. Conclusion: These results suggest that Complex GPB has an immune-enhancing effect, which can be used for immunomodulatory remedies.

Objectives: Resistance developed by micro-organisms against antibiotics is now a global threat. There is a need to look at alternate sources from which substances useful against various diseases can be obtained. Soil surrounding the medicinal plants possessing anti-microbial activity was screened to find soil bacteria capable of producing anti-microbial compounds and to isolate such compounds therefrom. Materials and Methods: Eleven anti-microbial medicinal plants from three regions of Saurashtra, Gujarat, India, were selected, and soil surrounding those plants was collected. Sixty-six isolates of actinomycetes were obtained from the soil samples. Cross streak, agar well, and disk diffusion methods were used for primary, secondary, and final screening, respectively, of the anti-microbial isolates. Results: Among all, the isolate from the soil surrounding Ocimum sanctum in the Gir-Somnath region was found to be most potent and subjected to thin-layer chromatography, bio-autography, bio-chemical tests, enzymatic activity, 16S rDNA sequencing, and gel electrophoresis, which was confirmed to be Streptomyces arenae, followed by optimization of various conditions. Isolation and characterization of the compound were performed by using ultra-violet, Fourier transform infrared, nuclear magnetic resonance, and mass spectroscopy. A novel compound was isolated and characterized, which showed potent anti-microbial activity as compared to reference standards by in vitro anti-microbial assay. Conclusion: Soil surrounding medicinal plants which themselves have well-known anti-microbial activity may be a rich source of actinomycetes, which may produce compounds having anti-microbial properties. This can be an emerging strategy for researchers and clinicians to explore soil bacteria for the isolation of biologically active compounds for the management of various diseases.

Background: In recent years, increasing resistance to synthetic agents, their long-term treatment, and lasting side effects, has faced many problems in treatment of leishmaniasis; so that finding a new high-efficacy antileishmanial drug with minimal side effects seems very necessary. This experimental study was aimed to evaluate the in vitro and in vivo leishmanicidal activity and cellular mechanisms of Astraglus spinosus methanolic extract (ASME) against Leishmania major infection. Materials and Methods: In vitro antileishmanial effect of ASME was evaluated on intracellular amastigotes of L. major in macrophage model. The effect of ASME on the NO production of macrophage cells was determined based on the Griess reaction for nitrites. Effect of ASME on the caspase-3-like activity of L. major promastigotes was performed according to the measuring the rate of color spectrophotometry. The 50% cytotoxic concentrations (CC₅₀) of the ASME on macrophages were measured to assess the cytotoxicity of ASME. In addition, in vivo effects of ASME were evaluated in infected BALB/c mice by measuring of the diameter of CL lesions and parasite load in the tested mice before and after 28 days of therapy. Results: The mean number of intracellular amastigotes of L. major significantly (P < 0.001) decreased with the IC₅₀ value of 36.9 ± 3.012 μg/mL and 44.3 ± 3.012 μg/mL for ASME and MA, respectively. Although more NO was produced by increasing the concentrations of the ASME, but, a notable rise was detected at the IC₅₀ (P < 0.001). ASME especially at the concentrations of ½ IC₅₀ and IC₅₀ significantly provoked the caspase-3 activation, by 10.3%, 25.6%, and 29.8%, respectively. The measured CC₅₀ value of ASME and MA was 463.3 μg/mL and 835 μg/mL, respectively. Treatment of the infected mice with various doses of ASME (50 and 100 mg/kg for 28 days), markedly declined the mean diameter of the CL lesions and parasite load in tested mice. Conclusion: Based on the obtained results, ASME can be considered as a promising herbal drug candidate for the isolation and production of a new alternative agent for CL treatment. As a result, this survey presented adequate results in the parasite eliminating in both in vitro and in vivo assay. Nevertheless, additional studies are needed to elucidate the accurate mechanisms of action of ASME and its effectiveness in clinical subjects.

Background: Asparagus racemosus has been attaining its importance due to its high medicinal and pharmaceutical value. This plant resulted in the depletion of its wild population in India. This plant has been placed in the nearly threatened and endangered category of the International Union for Conservation of Nature and Natural Resources (IUCN) in the states of Chhattisgarh, Madhya Pradesh, and West Bengal. The over-demand also poses the problem of adulteration in several ways. Objectives: Selection of elite germplasm for commercial cultivation was done by studying the variation of Shatavarin IV from different geographical regions. The development of simple and reliable high-performance thin layer chromatography (HPTLC) method and chromatographic profile was done for quality control. Materials and Methods: A total of 103 Asparagus racemosus root samples were collected from different agro-climatic zones of Odisha. The methanol root extracts were analyzed qualitatively and quantitatively using HPTLC. The pre-coated silica gel 60 F₂₅₄ plates were used for the development of chromatograms with ethyl acetate‒methanol-water (7.5:1.5:1, v/v/v) as a mobile phase. Results: The Shatavarin IV was detected at R_f of 0.40 ± 0.05 and showed maximum absorption at 425 nm. The method was validated in terms of linearity, limit of detection (LOD), quantification (LOQ), precision, stability, and recovery test. The amount of Shatavarin IV varied between 0.01-0.40% among different regions. Conclusion: The elite germplasms were identified at Ramagiri Hill sides of Gajapati district, Odisha, having Shatavarin IV content <0.39%. The HPTLC method used here was found to be reliable, precise, and can be used for the quality control assessment of Asparagus racemosus.

Background: Lung cancer exhibits extremely high mortality, and there is a pressing need to identify novel leads for the development of efficient chemotherapy for lung cancer. Thus, there is a need to identify novel and effective molecules for the management of lung cancer. Objectives: In the present study, the in vitro anticancer effects of globularifolin were evaluated against human lung cancer and attempts were made to explore the underlying molecular mechanisms. Materials and Methods: Cell viability and proliferation was determined by MTT, EdU, and colony formation assays. Cellular processes were detected by annexin V/PI, confocal microscopy and flow cytometry. Protein expression was detected by western blotting. Results: The results showed that globularifolin suppressed the growth and colony formation of the human lung cancer cells and exhibited an IC₅₀ of 8 μM against the HCC827 and SK-LU-1 lung cancer cell lines. However, the IC₅₀ of globularifolin was found to be comparatively higher against the normal MRC-5 cells. The globularifolin-induced inhibition of human lung cancer cells were found to be due to the induction of apoptosis, which was associated with upregulation of Bax, caspase-3 and caspase-9, and downregulation of Bcl-2. Further investigation of the underlying mechanism revealed that inhibitory effects of globularifolin were mediated through apoptotic induction driven through inhibition of PI3K/Akt and PINK/Parkin signaling pathways driven respectively through reactive oxygen species (ROS) production and mitophagy. Conclusion: Taken together, the results show that globularifolin exerts anticancer effects on human lung cancer cells via regulation of ROS and suppression of PINK1/Parkin mitophagy pathway.

Background: Muscle atrophy means a progressive decrease in muscle mass, strength, and quality and has a lot of discomfort in daily life. High-dose or continuous use of glucocorticoids (GCs), which are negative muscle regulators, led to the risk of muscle atrophy and weakness. Up to now, the effect and the underlying mechanism of Citri unshius Pericarpium (CP) on muscle atrophy have not been fully elucidated. Objectives: Accordingly, the current study was performed to evaluate the effects and the underlying mechanisms of CP on dexamethasone (DEX)-provoked muscle atrophy in C57BL/6 mice. Materials and Methods: The 8-week-old mice were treated once a day (DEX 20 mg/kg body weight, i.p.), and CP was administrated orally for 10 days. Then, we measured the body weight, swimming time, and muscle weight, and histological evaluation and western blot were performed. Results: CP improved muscle function decline by bettering the swimming time and muscle weight to some extent. Moreover, histological muscle damage induced by DEX was enhanced through CP treatment. CP treatment induced the reduction of ROS-related factors. CP showed a decrease in the protein expressions such as myostatin, Atrogin-1, and MuRF1 via the down-regulation of the phosphorylation of AMPK. Besides, in the CP group, muscle protein synthesis was increased by the PI3K/Akt/mTOR signaling pathway. Conclusion: Taken together, CP could be highly commercialized as a commercial material for functional food for the prevention and improvement of muscle loss, which is induced by muscle atrophy.

Background: This research explored the effects of vinorelbine combined with oxaliplatin on non-small cell lung cancer (NSCLC). Materials and Methods: NSCLC cells were treated with vinorelbine, oxaliplatin, and insulin-like growth factor-1 (IGF-1; phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) pathway activator), and transfected with ATP-binding cassette subfamily G member 2 (ABCG2) overexpression plasmid and silenced ABCG2 (siABCG2). Cell viability was detected by cell counting kit-8 assay, cell migration and invasion by the transwell assay, and cell apoptosis by flow cytometry. Levels of ABCG2, PI3K, phosphorylation (p)-PI3K, AKT, and p-AKT were evaluated through qRT-PCR and western blot. Results: Co-treatment with vinorelbine and oxaliplatin exerted synergistic effects in suppressing the proliferation, migration, invasion, and the phosphorylation of PI3K and AKT, and inhibiting ABCG2 expression while inducing apoptosis in NSCLC cells. Overexpression of ABCG2 reversed these effects on proliferation, migration, invasion, and apoptosis, while not affecting the PI3K/AKT pathway in NSCLC cells. IGF-1 inhibited the synergistic effect of vinorelbine and oxaliplatin on the NSCLC cells. Moreover, siABCG2 reversed the effects of IGF-1 on ABCG2 expression, and other cellular functions mentioned above in NSCLC cells receiving the co-treatment. Conclusion: A combination of vinorelbine with oxaliplatin represses the proliferation, migration, and invasion of NSCLC cells, and facilitates apoptosis via inhibition of the PI3K/AKT/ABCG2 axis.

Background: Epilepsy remains one of the leading neurological diseases. More than one mechanism exists for epilepsy. Inflammation and oxidative stress are amongst the mechanisms leading to epilepsy. In the current work, we evaluated whether Aegle marmelos (AM) is able to reverse epilepsy. Objectives: We induced epilepsy in zebrafish through the administration of pentylenetetrazole as a model organism. Materials and Methods: To assess the anti-epileptic potency of plant extract, behavioural changes were recorded and analysed by Behavioural Observation Research Interactive Software. Further, oxidative stress, glutathione peroxidase and lipid peroxidation (LPO) levels were estimated. Real-time polymerase chain reaction was performed to quantify the expression of inflammatory markers tumour necrosis factor-alpha (TNFα) and interleukin 1 beta (IL-1β). Results: Epilepsy-induced animals displayed a pronounced behavioural change in swimming patterns and preference for darkness and stayed longer periods at the bottom of the tank. On the other hand, plant extract treated animals reversed the behavioural alterations induced by epilepsy. Oxidative stress was seen in animals induced with epilepsy and the same was reversed in animals that were treated with plant extract. LPO was elevated in epilepsy whereas plant extract reduced the same. Inflammatory markers such as TNFα and IL-1β levels were higher in epileptic animals whereas their expression levels were reduced by treatment with the plant extract. Histopathological examination also revealed the inflammatory reaction in the brain induced by epilepsy. Conclusion: Treatment with the plant extract restored the tissue architecture to normal. Overall, there was a major reversal of behaviour, oxidative stress and inflammation upon treatment with the plant extract indicating the anti-epileptic potential of AM.

Background and Objectives: Pain or nociception is a devastating experience that alters both the physiological and psychological status of individuals. A variety of chemical and physiological tissue damage and physical parameters like temperature, pressure, etc. induce pain. Clinically, pain is managed through the administration of antinociceptive drugs. In the current work, the antinociceptive potential of Mentha piperita leaves against pain in the zebrafish model has been explored. Materials and Methods: Formalin was injected in the tail of the zebrafish to induce pain and was exposed to 5, 10, 15, and 20 mg of the extract. The behavioral pattern was analyzed using the Behavioral Observation Research Interactive Software (BORIS). Gene expression and enzymatic studies were performed to evaluate inflammation, oxidative stress and the antioxidant ability of M. piperita. Results: In the open field test, animals treated with the leaf extract restored their activity to normal in a dose-dependent manner. Pain-induced animals inclined toward darkness, whereas the treated animal preferred to stay in the light. Furthermore, behaviors such as freezing, inaction, and altered swimming patterns were all restored to normal conditions. The color change in the tail was prominent in pain-induced animals, whereas it was restored to normal in the treated group. The higher dose of the plant extract was effective in downregulating the expression of inducible nitric oxide synthase (iNOS) and tumor necrosis factor alpha (TNF-α), thereby impairing the neuroinflammation. The plant extracts reduced reactive oxygen species (ROS) and nitric oxide levels, whereas it increased reduced glutathione levels. Conclusion: The current work establishes that Mentha piperita is effective in ameliorating pain in a zebrafish model.

Aim: The study herein was performed in view of prospecting 15 species and one variety of Barleria in comparison to Adathoda vasica for phytochemical composition, and antioxidant activities. Materials and Methods: In addition to this, the study also quantifies two anti-bronchial (vasicine, vasicinone) and one anti-cancer (betulin) compounds using reversed-phase high-performance liquid chromatography analysis. Results: Samples showed betulin ranging from 7.700 to 73.447 mg/g, vasicine from 0.092 to 3.710 mg/g, and vasicinone from 0.005 to 2.752 mg/g. Principal component analysis and hierarchical cluster analysis results signified the division of seventeen samples tested, into two major clades (based on a similarity range 57.83–96.60%). Conclusion: Both phytochemical analysis and antioxidant activities showed variation in the samples tested. Barleria grandiflora had higher content and better activities among the species.

Background: Antibiotic resistance in bacteria mediated by New Delhi Metallo-β-lactamase (NDM) is a global threat to human health with an enormous economic burden. NDM can hydrolyze all the β-lactam core-containing antibiotics including carbapenems, which are regarded as last resort antibiotics. Materials and Methods: A library of Abyssomicins was virtually screened to identify novel non-β-lactam ring-containing inhibitors of NDM-1. Different computational approaches such as molecular modeling, virtual screening, molecular docking, molecular dynamics simulation, ADMET profiling, and free energy calculations were utilized for this purpose. Results: Virtual screening and ADMET profiling shortlisted Abyssomicin W and Neoabyssomicin B as the most promising candidate molecules. An in-depth analysis of protein-ligand interactions by molecular docking revealed that both ligands bind the active site of NDM-1. The identified inhibitors interacted with key catalytic residues as well as other residues around the active site of NDM-1. Hydrogen bonding and hydrophobic interactions played a significant role in stabilizing the protein-inhibitor complexes. The docking energy of NDM-1-Abyssomicin W, and NDM-1-Neoabyssomicin B complexes were − 9.6 kcal/mol and − 9.5 kcal/mol, respectively, which were higher than NDM-1-Methicillin (control) complex (−7.3 kcal/mol). Molecular dynamics simulation and free energy calculations by MM-PBSA also confirmed the stability of NDM-1-Abyssomicin W, and NDM-1-Neoabyssomicin B complexes. Conclusion: The findings of this study suggest that Abyssomicins serve as potential inhibitors of NDM-1. However, these results need to be validated in vitro and in vivo. This study may serve as a basis for further developing Abyssomicins as novel inhibitors of β-lactamases.

Background: In this study, we elucidated the neuroprotective effect of dioscin, a plant-derived steroidal saponin utilized in Chinese medicine to treat various diseases. To achieve this, we established the aluminum chloride–induced Alzheimer's disease rat model. Materials and Methods: Young, healthy, male Wistar rats were grouped into five: control, Alzheimer's disease–induced, Alzheimer's disease–induced and treated with 10 mg/kg dioscin, Alzheimer's disease–induced and treated with 20 mg/kg dioscin, and Alzheimer's disease–induced and treated with standard neuroprotective drug donepezil (1.5 mg/kg/day) by oral route. After completion of the treatment period, the animals were subjected to behavioral, biochemical, molecular, and histological analyses. Morris water maze and open-field tests were conducted to analyze the behavior of the animals. Acetylcholinesterase, glutamate, neurotransmitters, and oxidative stress markers were quantified to assess the biochemical changes in Alzheimer's-induced and dioscin-treated rats. Furthermore, quantitative polymerase chain reaction (qPCR) analysis was done to assess the mRNA expression of proinflammatory genes, and the neuroprotective effect of dioscin was confirmed with histopathological analysis of hippocampal region. Results: Aluminum chloride–induced neurodegeneration in rats was evident with behavioral, biochemical, molecular, and histological analyses. The results of behavioral analysis revealed that dioscin treatment increased the memory in Alzheimer's-induced rats. It also decreased the oxidative stress markers, and increased the level of norepinephrine and dopamine neurotransmitters involved in cognition. The mRNA expression of proinflammatory cytokines was significantly decreased in Alzheimer's-induced dioscin-treated rats. Histopathological analysis revealed that dioscin protects the hippocampus from aluminum chloride–induced neurodegeneration. Discussion: Overall our results confirmed that dioscin is a potent neuroprotectant and can be utilized to treat Alzheimer's disease in the future.

Background: In the medical and pharmaceutical fields, man has successfully used natural-origin things for millennia. Polymers obtained from plant sources have flashed much consideration in current eras owing to their various pharmaceutical properties, such as gelling agents in gels. Also incorporated in cosmo products, fabrics, dyes, and paper manufacture as binders, diluents, disintegrating agents, thickeners in oral fluids, protective colloids in suspensions, and bases in a suppository. Objectives: Natural polymers are chosen over semisynthetic and synthetic excipients due to their lack of toxicity, low cost, availability, soothing action, and nonirritant nature, as well as their ability to undergo a variety of chemical modifications. However, they have some drawbacks, including microbial contamination, lot to lot variation, reduced viscosity throughout storage, inappropriate mechanical properties, low storage, and an uninhibited rate of hydration. Current research work aims to chemically modify xanthan gum and study its impact on the formulation of topical gels. Materials and Methods: Chemical modification of xanthan gum is conceded by using a thiol-esterification reaction with thioglycolic acid (TGA) in the presence of hydrochloric acid. Microwave irradiated thiolated xanthan gum was also willing by further irradiation of thiolated xanthan gum by microwave using 750 W frequency. Topical drug delivery of lornoxicam release applications of thiolated and microwave irradiated xanthan gum are comparatively evaluated with pure xanthan gum polymer by formulating gel incorporated lornoxicam as a drug. Results: The physical properties of gel-like physical evaluation and viscosity results are within official limits. Formation of gel with idyllic properties. Determination of drug content, spreadability, and extrudability of modified xanthan gum was high compared with native xanthan gum. In vitro drug release study showed that prepared gel sustains the drug release for 24 h. Conclusion: Thiolation and irradiated xanthan gum sustain the release of lornoxicam over a prolonged period and might be a promising lornoxicam carrier in topical drug delivery to enhance antinociceptive and anti-inflammatory efficiency.

Background: Cancer, a deadly disease seems to be a major concern for the global medical field. Controlling the spread of disease is not only a problem for under developed countries but also the developed countries face a lot of concern in diagnosing and treating cancer. Among various cancer types lung cancer mortality incidence seems to be a global threat. Surgery and Chemoradiation therapies are mostly prescribe for lung cancer patients which renders post treatment side effects. Hence, treating cancer with alternative medicine tends to be a choice at present. Materials and Methods: In this study, we induced lung cancer in healthy Swiss albino mice by treating with polycyclic aromatic hydrocarbon carcinogen benzo(a)pyrene. The treatment was carried out for a period of 18 weeks; both pre supplementation and post supplementation of prunetin was administered to the mice. Results: The changes in hematological and immunological parameters induced by prunetin in lung cancer induced mice were assessed. Oxidative stress in white blood cells which plays a key role in lung cancer induction was analyzed. The therapeutic of efficacy of prunetin was evaluated by estimating the levels of metabolizing enzymes in cancer induced mice. The anticancer effect of prunetin was assessed by estimating the tumor marker CEA and anti-inflammatory markers. It was further confirmed with histopathological analysis of lung tissue. Our results had proven that prunetin significantly increased the white blood cell count, phagocytic, and activity index in lung cancer induced mice. It also decreased the levels of immunoglobulins and oxidative stress in white blood cells. Prunetin treatment decreased the levels of metabolizing enzymes, tumor marker CEA, and proinflammatory markers in cancer induced mice which proven the anticancer efficacy of prunetin. It was further confirmed with histopathological analysis of lung tissue. Conclusion: Over all our results authentically confirms prunetin is a potent anticancer drug to treat lung cancer.

Background: Osteoarthritis (OA) is a major bone-related disease, which is characterized by joint deterioration, bone destruction, and whole joint damage, leading to permanent disability. The present study aimed to assess the anti-inflammatory and anti-osteoarthritic effects of esculetin in a monosodium iodoacetate (MI)-induced OA model in Sprague-Dawley (SD) rats. Materials and Methods: The anti-osteoarthritic efficacy of esculetin was determined in an MI-induced OA animal model. Rats were divided into the following groups: group I: normal control (saline), group II (MI only treated), group III (MI + esculetin), and group IV (MI + indomethacin). The potent outcome of esculetin treatment was assessed through its effects on the proinflammatory cytokines' levels, weight-bearing distribution, and histopathologic observation. Anti-inflammation efficacy of esculetin was evaluated in lipopolysaccharide (LPS)-induced Ralph and William's cell line 264.7 (RAW 264.7) cells. Results: In vitro results showed that esculetin has an anti-inflammatory efficacy through lessening the production of nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1β, and IL-6 in LPS-induced RAW cells. Furthermore, esculetin also reinforced the retrieval of hind limb weight-holding capacity, downregulated the generation of inflammatory mediators and proinflammatory cytokines, and shielded or protected the cartilage tissue from damage in the OA model SD rats. Conclusion: Esculetin seems to be a beneficial therapeutic compound for handling OA and OA-based disease conditions.

Objectives: Formononetin is a natural isoflavone compound and has been reported to have anti-cancer properties. However, the mechanism by which formononetin inhibits hormone-resistant prostate cancer (PCa) has not been reported. The present study investigates whether and how formononetin inhibits the epithelial-mesenchymal transition (EMT) of hormone-resistant PCa cell lines. Materials and Methods: The proliferation, migration and invasion of PCa cells were analyzed using RTCA system and CCK8 assays. Expression of EMT-related markers and MAPK pathway were analyzed using quantitative reverse transcription PCR (RT-qPCR), immunofluorescence staining and immunoblotting. Gene expression profile was observed by RNA-sequence analysis, and the EGR1 expression analysis in human PCa tissue was referred to the Cancer Genome Atlas database and The Human Atlas Database. Results: Formononetin inhibited the rate of proliferation, migration, and invasion of PCa cells, increased protein levels of E-cadherin, reduced protein levels of fibronectin and the phosphorylated ERK1/2 and JNK, as well as the mRNA levels of the fibronectin, slug and snail. Formononetin remarkably upregulated EGR1, as confirmed by bioinformatics analysis, RT-qPCR and western blotting. Furthermore, the EGR1 expression was lower in human PCa tissue versus its control, and in higher Gleason grade versus the lower Gleason grade PCa. Conclusion: Formononetin could ameliorate tumor aggressiveness via reversing the EMT of hormone-resistant PCa cells. The potential mechanisms involved are inactivation of MAPK signalling and subsequent upregulation of EGR1 expression.

Background: Euphorbia is one of the plants most used by herbalists and therapists in Morocco. Objectives: The aerial part of two plant samples (Euphorbia resinifera and Euphorbia officinarum) collected in Morocco was examined for the solvent effect, extraction time, and plant concentration in order to determine the best extraction conditions. Materials and Methods: To achieve this goal, a response surface methodology (RSM) using a full three-level factorial design was used to optimize the conditions for the extraction of antioxidants and α-glucosidase inhibitors. Temperature, time, and plant-to-solvent ratio (PSR) and their linear and quadratic interactions on TPC (total phenol concentration), TFC (total flavonoid concentration), DPPH (2,2-diphenyl-1-picrylhydrazyl) trapping activity, and α-glucosidase inhibiting activities were studied. Results: According to desirability functions, the optimum operating conditions to achieve a higher extraction yield of phenols and higher antioxidant and anti-diabetic activity were found by using extraction during 60 min at 30°C using a PSR of 20 mg/mL, whereas a longer extraction time (270 min) was needed for E. resinifera and a higher extraction temperature (50°C), with a lower PSR (10 mg/mL) for E. officinarum. Conclusion: In order to find the best conditions to extract secondary metabolites with biological activity and application in phytotherapy, the appropriate solvent generally used by populations, water in this case, should be used, but the best extraction conditions have to be found in order to enhance the pharmacological actions.

Background: Green chemistry is widely accepted phenomenon to synthesize iron oxide nanoparticles (Fe-NPs) used in several biomedical and technological applications. Metal oxide nanoparticles are useful in biomedical, clearing environmental pollutants, enzyme immobilization, etc., Objectives: The synthesis of Fe-NPs using Sphagneticola trilobata leaf extract using ferric chloride solution and its biological assessment. Materials and Methods: The present study involved the synthesis of Fe-NPs using S. trilobata leaf extract using ferric chloride solution by the co-precipitation method. The synthesized nanoparticles were characterized for Fourier-transform infrared spectroscopy, scanning electron microscopy, powder X-ray diffraction spectroscopy, particle size analysis, and magnetization studies. The nanoparticles were biologically evaluated for microbiological, antioxidant, and in vitro cytotoxicity activity. Results: Magnetic nanoparticles were appeared in dark brown color. The change in color might result due to the presence of polyphenols in S. trilobata leaf extract. The characterization studies confirmed morphology, shape, and size of the nanoparticles. The mean diameter of Fe-NPs and S. trilobata-Fe-NPs was found to be 42.2 ± 2.6 and 62.54 ± 2.01 nm, respectively. Magnetization studies of nanoparticles revealed ferromagnetic behavior with the saturation magnetization at 57 emugm⁻¹. S. trilobata-Fe-NPs showed significant antibacterial action against Staphylococcus aureus and Bacillus subtilis by the well-diffusion method. Antioxidant activity of S. trilobata-Fe-NPs exhibited 65.78% inhibition in comparison with ascorbic acid. The cytotoxicity assay of S. trilobata-Fe-NPs on HCT-15 colon adenocarcinoma cells showed significant anticancer activity (56.44%) cytotoxic inhibition. Conclusion: Green synthesis-mediated S. trilobata-Fe-NPs appeared to produce significant antimicrobial and anticancer potential.

Background: β-amyloid (Aβ) deposition and tau protein abnormality are the major pathogenesis of Alzheimer's disease (AD). Autophagy is contributed to eliminating the misfolded proteins or organelles and alleviated cellular injury. Our pre-experimental findings showed that bornyl acetate (BA), the main component of the volatile oil of Amomum villosum, has a neuroprotective effect in okadaic acid (OA)–induced PC12 cells. However, the protective mechanism of autophagy that BA relieves OA-induced cellular injury is still unclear. Objectives: The purpose of our experiment was to elucidate the mechanism of treatment of AD. Materials and Methods: To explore how BA has therapeutic effects on AD, a model of OA-induced PC12 cells was established. The OA modelling induction and BA treatment in cells were evaluated by LDH and CCK-8 methods; ELISA assay was used to detect the tau hyperphosphorylation (p-tau), Aβ₄₂ and β-secretase levels; The expression of p-Akt and p-mTOR were detected by western blot analysis; and immunohistochemical, immunofluorescence methods and western blot analysis were used to detect Beclin-1 expression. Autophagosomes in each group were observed by transmission electron microscopy (TEM). 175 nM OA for 48 hr was applied to OA modelling induction. Results: Compared to the control group, the OA-induced AD model cells displayed higher levels of p-tau, Aβ₄₂ and β-secretase (P < 0.01), suggesting that the AD model was successfully established. Compared to the OA model group alone, p-tau, Aβ₄₂ and β-secretase levels and autophagy promoter Beclin-1 expression decreased significantly in the BA group (P < 0.05), whereas p-Akt and p-mTOR increased (P < 0.01). Conclusion: BA exhibited a neuroprotection effect against OA-induced cellular injury in the AD model by suppressing the Beclin-1-dependent autophagy pathway, indicating that BA might be an appealing potential strategy to treat AD.

Objectives: Hyphaene thebaica Mart. (locally known as Doum) is a well-known tropical plant and is traditionally used for the treatment of gastrointestinal tract (GIT) ulcers. The current study aimed to investigate the antioxidative and gastroprotective properties of H. thebaica fruit rind extract against experimentally ethanol-induced gastric ulceration in Sprague–Dawley rats. Materials and Methods: Two experiments were carried out: first, with 20 animals for dose selection and safety of the extract; then, a second with 30 animals for the gastric ulcer model. The two selected doses of H. thebaica extract (250 and 500 mg/kg) and antiulcer drug (omeprazole: 20 mg/kg) were administered through oral gavage for 2 weeks prior to ulcer induction. Acidity, mucus weight, ulcer area, and histopathology were used to assess the gastroprotective effects of H. thebaica extract. The antioxidative properties were assessed using the lipid peroxidation assay, non-protein thiol levels, superoxide dismutase activity, nitric oxide assay, and immunohistochemical staining of mitochondria-regulated apoptosis proteins such as Bax and Bcl-2. Results: In the current study, no in vivo toxicity of H. thebaica extract was observed. In the gastric model, preadministration of H. thebaica resulted in a significant reduction in ulcer area and mucus weight in a dose-dependent manner. Moreover, gross and histological findings confirmed the gastroprotective properties of H. thebaica extract. Quantitative assessment of microscopic lesions revealed a significant difference between the groups. These properties were observed to be mediated through the modulation of oxidative stress. H. thebaica modulated the Bcl-2 and Bax proteins and inhibited apoptosis. Conclusion: The gastroprotective properties of H. thebaica nominate it as a potential nutraceutical candidate.

Background: Licorice, one of the most commonly used phytomedicines in Asia, has detoxication and anti-oxidation properties. Due to the exhausted wild resources, licorice is obtained mainly from cultivation, and the suitable conditions under cultivation lead to a serious decline in quality. Objectives: To establish a method to improve the quality of cultivated licorice by environmental stress. Materials and Methods: The fresh roots of Glycyrrhiza uralensis were soaked in 5%, 10%, and 20% polyethylene glycol-6000 (PEG) for 4 days to construct a physiological state of drought. The changes in hydrogen peroxide (H₂O₂) and enzymes related to secondary metabolism in the treated licorice were observed, and their detoxication and anti-oxidation were verified by reducing aconite-induced cardiotoxicity and free radical scavenging rate experiments. Results: The H₂O₂ content notably boosted, and gene expression and allosterism of enzymes related to secondary metabolism such as 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGR), squalene synthase (SQS), β-amyrin synthase (β-AS), and phenylalanine ammonia-lyase (PAL) were promoted, and enhance their activities. Under a drought stress of 10% PEG, secondary metabolites significantly increased, and the effectiveness of the drug also intensified. For the myocardial injury caused by aconite, compared with the untreated, creatine kinase (CK), cardiac troponin-T (cTn-T), and lactate dehydrogenase (LDH) decreased by 9.3%, 14.6%, and 6.3% in the 10% PEG. Meanwhile, the clearance rates of 1,1-diphenyl-2-picryl hydrazyl radical (DPPH·) and hydroxyl radical (OH·) heightened by 7.5% and 13.1%, separately. Conclusion: Drought stress can greatly increase the secondary metabolites of licorice and enhance its detoxication and anti-oxidation.

Background: Because of the certain overlapping of the genus Bupleurum, the identification features are few and the delimitation among species is difficult, Bupleurum taxonomy is under frequent revisions. In this research, hierarchical cluster analysis based on morphology characters is done to establish a dendrogram and illustrate the taxonomic relationship among Bupleurum species (Apiaceae) from China. Materials and Methods: The fruit morphology of 17 Bupleurum taxa from China was investigated with a scanning electron microscope (SEM) and optical microscopy (OM). The hierarchical cluster analysis based on 41 morphological character states belonging to 9 characters was done to establish a dendrogram and illustrate the taxonomic relationship among the studied species of Bupleurum (Apiaceae) from China. Results: Based on fruit morphological characteristics, it is possible to separate the Bupleurum species into the following four groups. The research indicates that the classification of Bupleurum in China is closely related to geographical distribution, which is divided into Southwestern and Northeastern Bupleurum. Conclusion: The fruit morphological characteristics are variable enough to distinguish species between Bupleurum, such as fruit length, width, shape, colour, the curvature of the outer periclinal and so on in this article. Morphological characters could be probably the taxonomical standard of intraspecific and interspecific delamination, which is significant for understanding the classification of Bupleurum.

Background: Hepatocellular carcinoma (HCC) is the utmost familiar form of liver distortion, a prevalent high incidence that ensures the fifth most recurrent primary malignancy around the world. Objectives: To investigate the chemopreventive and apoptotic effect of vernodalin in diethylnitrosamine (DEN) induced HCC. Materials and Methods: Rats were distributed into four sets of experimental animals every six rats: control group, DEN alone, DEN + vernodalin, (10 mg/kg bw), and vernodalin (10 mg/kg bw) alone. The analysis was conducted to measure the liver and body weight. The assay was designed to analyze hepatic toxic markers, antioxidants, lipid peroxides, and inflammatory cytokines, in addition, histopathology and western blot analysis were also conducted. Results: DEN with vernodalin diminished weight of the body and antioxidant level whereas raised the liver weight, toxic marker concentration, oxidative stress and cytokines. Vernodalin also prevents oxidative stress, lipid peroxides level, toxic markers, pro-inflammatory, and cytokines. Vernodalin suppressed the P13K/AKT signaling that enhances apoptosis and maintains the morphology of cells. Conclusion: The results indicate that vernodalin exerts a promising chemopreventive and apoptotic natural agent for DEN-induced HCC in rats.

Background: Vicia amoena Fisch. var. Angusta. Freyn, also known as tuogucao, is a traditional medicinal plant in China. Early researches of V. amoena Fisch. var. Angusta. Freyn were focused on biosystematical studies, and only a few were concerned with chemical constitutes and bioactivities. Objectives: We aimed to isolate chemical constitutes in aerial parts of V. amoena Fisch. var. Angusta. Freyn and study the immunomodulatory activity of these chemical components via immune cells. Materials and Methods: Polysaccharide was obtained via hot water extraction and alcohol precipitation at different concentrations. Individual compounds were obtained after several chromatographic techniques, and the structures of all individual compounds were characterized by physical-chemical process, ¹H, ¹³CNMR, UV techniques and by comparing their NMR data with those discovered in the literature. In vitro immunological activity of tested samples was evaluated using MTT assay, NO assay, scratch assay, phagocytosis assay and proliferation assay of spleen lymphocytes in mice. Results: The four polysaccharide sites including 30%, 50%, 70% and 90% were obtained. Eight pure chemical constitutes were isolated and identified as: isoquercitrin, quercetin, quercitrin, hyperin, chlorogenic acid, kaempferol, kaempferitrin and wogonin. Four polysaccharide sites and individual compounds displayed different levels in improving proliferation and phagocytosis of RAW264.7 macrophages and lymphocytes. Conclusion: Compounds were isolated from the herb that possessed immunomodulatory function, and it is likely that the extract from the herb via the TLR-4-mediated signalling pathway to achieve immunomodulatory effects provided a scientific rationale of the traditional uses of the herb.

Background: Non-small cell lung cancer (NSCLC) is often harmful to human health. It is common in people who smoke or smoke passively, and the affected people are older and find it difficult to recover. The main treatment methods are surgery, radiotherapy and chemotherapy. This paper aims to study the effects of different configurations of ginsenoside Rh2 on NSCLC 95D and NCI-H460 cells. To provide clinical value for the treatment of NSCLC. Materials and Methods: CCK-8 was used to determine the proliferation of 95D and NCI-H460 cells with different concentrations of ginsenoside Rh2 (0, 0.05, 0.1 and 0.2 mg/mL), and IC₅₀ was calculated. Cell cycle and apoptosis were detected by immunofluorescence staining and flow cytometry. Results: 20(R)-ginsenoside-Rh2 and 20(S)-ginsenoside-Rh2 inhibited the proliferation and colony formation of NSCLC 95D and NCI-H460 cells induced cell cycle arrest in a time and dose-dependent manner in G1/S and promote apoptosis. Conclusion: Ginsenoside Rh2 induces antitumour activity by inhibiting proliferation and colony formation of 95D and NCI-H460 cells and inducing cell cycle arrest and apoptosis. Comparing the two configurations of ginsenoside Rh2, the effect of 20(R)-ginsenoside-Rh2 is stronger.

Background: Galangin (Gal) is a natural active flavonoid compound separated from the roots and rhizomes of Alpinia ofcinarum Hance. Modern pharmacological studies have shown that Gal has a variety of biological activities such as antitumor, antifungal, antibacterial, antiinflammatory, antiischemic stroke, suppressing vitiligo and Alzheimer's disease, etc., Objectives: In this study, our primary goal was to prepare a galangin self-microemulsion drug delivery system (Gal-SMEDDS) and evaluate the effect of free Gal and Gal-SMEDDS on the pharmacokinetic parameters of SD rats, and the protective effect of oxidative damage on human embryonic lung fibroblasts (HFL1) cells in vitro. Materials and Methods: Gal-SMEDDS was prepared by ethyl oleate, Cremophor CO 40, and PEG-400, and then evaluated by morphology, particle size, zeta potential, polydispersity index, entrapment efficiency, and the pharmacokinetic parameters. The oxidative damage model of HFL1 cells was established by the stimulation of H₂O_2. And the antioxidant effect of Gal-SMEDDS on HFL1 cells was evaluated by ROS fluorescence assay kit and Annexin V-FITC/7-AAD double staining cell apoptosis detection kit. Results: The average particle size of Gal-SMEDDS was approximately 21.33 nm, the polydispersity index was 0.096, the zeta potential was − 4.09 mV, and the entrapment efficiency was 96.74%. Compared with free Gal, the release of Gal-SMEDDS was improved in vitro release experiment. Cell experiments showed the anti-oxidant effect of Gal-SMEDDS was better than free Gal. In vivo pharmacokinetic experiments showed that the pharmacokinetic parameters of Gal-SMEDDS were better than that of free Gal. Conclusion: The SMEDDS effectively increased the oral bioavailability of Gal and improved its pharmacokinetic parameters, which were conducive to the anti-oxidant effect of Gal.

Background: Disruptive cholinergic neurotransmission and abnormal cognitive functions were the characteristics of Alzheimer's disease leading to abnormal brain–liver inflammatory responses. Oxidative stress was prominent in Alzheimer's disease-related neurodegeneration leading to inflammatory communication through altered cytokines between the brain and peripheral organs, mainly the liver. Objectives: Current study would demonstrate the healthier effect of N-acetyl cysteine and rutin against AD in rats. Materials and Methods: Experimental Wistar rats have been grouped appropriately for the administration of scopolamine and treatment using a combination of N-acetyl cysteine and rutin for 10 weeks. Results: In our study, scopolamine (2 mg/kg b.w.i.p.) administered Alzheimer's disease model in Wistar rats showed abnormality in behavioural changes (Morris water maze test), pro-inflammatory cytokines, decreased activities of enzymatic antioxidants, decreased reduced glutathione content, elevated brain oxidative stress markers, increased amyloid-beta, elevated acetylcholinesterase, elevated butyrl cholinesterase, increased phosphorylated tau protein, increased GSK-3β, BDNF, altered expressions of β-secretase, NADPH oxidase 2, and Nrf2 genes in brain, and augmented oxidative stress in liver affecting brain–liver axis. Oxidative stress in the liver was evident through decreased activities of enzymatic antioxidants, decreased glutathione content, and elevated liver oxidative stress markers. Conclusion: Treatment with N-acetyl cysteine with rutin showed protective efficacy by modulating the pathways related to Nrf2, NOX-2, and BACE1 genes in combating these abnormalities in rats. Modulation in the gene expressions in the brain tissue showed direct evidence of the drug's neuroprotective efficacy for future therapeutic strategies in scopolamine-induced Alzheimer's disease.

Background: The compound volatile oils (CVOs) from Rimulus cinnamon and Angelica sinensis are commonly used together in many compounds. The unstable and volatile nature of CVOs limits their application. Objectives: The objective of this study was to prepare and evaluate the CVOs microcapsules. Materials and Methods: Chitosan (CS) and sodium alginate (SA) were used as composite wall materials to encapsulate CVOs using the complex coagulation method. The microcapsules were characterized by scanning electron microscopy (SEM), Fourier transforms infrared spectroscopy (FT-IR), X-ray diffraction (XRD), and thermogravimetric (TG). The drug loading (DL%), encapsulation efficiency (EE%), in vitro release behavior, and stability of the microcapsules were studied. Results: DL (%) and EE (%) of microcapsules were 22.60 ± 0.02% and 72.55 ± 0.04%, respectively. SEM showed that microcapsules were spherical and the surface was not smooth, with lump-like folds, minor dents, and cracks. FT-IR and XRD confirmed the inclusion of CVOs within microcapsules. TG analyses indicated that the microcapsules possess strong heat resistance. Swelling experiments reflected the response of microcapsules to different pH values. Swelling ratios in acidic, neutral, and alkaline media were 2.47 ± 0.07, 1.75 ± 0.03, and 1.04 ± 0.01, respectively. The in vitro release behavior of the CVOs microcapsules demonstrated a sustained release property. Furthermore, the microcapsules had excellent stability, and the DL was 15.12% after storage at 25°C for 60 days. Conclusion: The CVOs microcapsules prepared by SA and CS not only showed regular appearance, uniform particle size, responsiveness to different pH values, and sustained release properties in vitro but also exhibited good DL performance and stability, which was helpful for further development into the various specific formulation.

Background: Osteoporosis is a multifactorial bone disease that progresses without notice until attaining a chronic stage. It causes socio-economic burden, higher mortality rate, and increased bone fractures. Ononin, an isoflavone present in numerous herbal Chinese medicinal formulations such as Astragali radix and red clover, has been reported to have potent therapeutic properties. Objectives: In this study, we aimed to evaluate the efficacy of ononin against ovariectomy(OVX)-induced osteoporosis in female rats. Materials and Methods: Ovaries were removed surgically to develop the animal model of OVX. Animals (n = 4) were grouped into control, OVX induced, OVX rats treated with ononin (10 mg/kg body weight), and OVX rats treated with 20 mg/kg body weight of ononin. Results: OVX rats treated with ononin showed improved body weight, uterus index, inflammatory and bone markers, serum lipid profile, and calcium level. Ononin downregulated the expression of estrogen (E2), acid phosphatase, and bone gamma-carboxyglutamate (Gla) protein in OVX rats. It also restored trabecular area and thickness, which was evidenced by histomorphometrical and histopathological examination. Ononin attenuated the expression of alkaline phosphatase, osterix, and Runx2 in OVX rats. Conclusion: In summary, our results suggest that ononin could be used as an ideal therapeutic agent to prevent and treat bone-associated disorders.

Background and Aim of the Study: Antiplatelet drugs available on the market have multiple side effects and pose drug interactions, which actuated the researchers to consider better alternatives by using plant extracts. It is well known that medicinal plants possessing anti-inflammatory action usually have antiplatelet aggregation and anticoagulation potential. Therefore, it was hypothesized that resins of Astragalus sarcocolla Dymock possessing anti-inflammatory properties would also manifest antiplatelet aggregation and anticoagulant properties. The objective of the present study was to assess the antiplatelet and anticoagulant effects of four different concentrations of ethanolic extracts (100, 80, 60, and 40%) of A. sarcocolla Dymock. Materials and Methods: Four different doses of A. sarcocolla Dymock extract (2.5, 5, 7.5, and 10 μg) were prepared for assays. In vitro antiplatelet and anticoagulant effects using human blood were studied in 42 healthy males aged between 25 and 35 years. Aspirin (0.5 μg) was used as the positive control for the antiplatelet aggregation assay. Furthermore, molecular docking was done to check the interaction between plant constituents and P2Y₁, P2Y₁₂, and phosphoinositide 3-kinase (PI3K). Chlorogenic acid, ferulic acid, and cinnamic acid were quantified using high-performance liquid chromatography (HPLC)-UV in ethanolic extract. Results: An HPLC analysis of the ethanolic extract of A. sarcocolla Dymock revealed the presence of chlorogenic acid, ferulic acid, and cinnamic acid. All the extracts of A. sarcocolla Dymock significantly inhibited aggregation of platelets against all three agonists' epinephrine (EPI), adenosine 5' diphosphate (ADP), and collagen (COL) in a concentration-dependent manner. All the extracts significantly affected prothrombin time and activated partial thromboplastin time in a concentration-dependent manner. Molecular docking showed strong interactions of chlorogenic acid and ferulic acid with P2Y₁, P2Y₁₂, and PI3K. Conclusion: The data of the present study revealed the inhibition of platelet aggregation and anticoagulation potential of active compounds of Astragalus sarcocolla Dymock, together with their significant interaction profile with the selected therapeutic targets.

Background: Atractyloidis rhizome (cangzhu) was an important herb widely used in clinical application with different products, such as crude cangzhu (CZO), cangzhu pieces (CZP), bran frying cangzhu (BCZ), and Jiao cangzhu (JCZ). Objectives: To unscramble the phytochemical profiling of CZO, CZP, BCZ, and JCZ. Materials and Methods: In this paper, ultra-high performance liquid chromatography combined with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS) technology and multivariate data analysis methods such as principal component analysis (PCA) and orthogonal partial least squares discrimination analysis (OPLS-DA) were employed to screen the different components among CZO, CZP, BCZ, and JCZ. Results: As a result, CZO, CZP, BCZ, and JCZ were separated obviously in PCA plot, and a total of 101 components were identified successfully. Among them, 54 chemical markers, including isoleucine, citric acid, L-phenylalanine, atractyloside A, etc., discriminating among CZO, CZP, BCZ, and JCZ, were screened out. Conclusion: UHPLC-Q-TOF-MS/MS technology combined with PCA and OPLS-DA could be successfully applied in clarifying the different ingredients between crude drug and its processed products of traditional Chinese medicine (TCM). Importantly, the established method revealed the reasonability of processing theory in TCM.

Background: The mechanism of a water-soluble extract of safflower on chronic alcoholic liver injury is still unclear. Objective: To study the mechanism of water extract of safflower on chronic alcoholic liver injury. Materials and Methods: The rats were separated in a random manner into five cohorts, namely, the normal control group (NC), alcoholic fatty liver disease model (ALD), ALD rat model treated with the water-soluble extract of safflower (30 g/kg), ALD rat model treated with the water-soluble extraction of safflower (40 g/kg), and ALD rat model with the water-soluble extract of safflower (50 g/kg). Rats in each group were anaesthetized with 2% isoflurane and euthanized by cervical dislocation after 1, 7, 14, 21, and 28 days treatment. Proteomic analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG analysis), immunohistochemistry (IHC), and western blot were employed for comparing the variations across model cohort and control cohort at the tissue level, protein level, and molecular level, respectively, and for probing potential prophylactic role/mechanisms for safflower extract on alcoholic liver injury. Results: KEGG analysis found that the differentially expressed genes in the alcoholic liver injury model were closely associated with alcoholism, inflammation, and serotonergic synapses. Furthermore, with the administration of water-soluble extract of safflower, these hepatic function indices and liver steatosis lesions were alleviated in both time and dose-dependent manners. Importantly, IHC analysis discovered that proteins disulfide isomerase A3 (PDIA3) and Trk-fused gene (TFG) were highly increased in the liver of the rats. Furthermore, the water-soluble extract of safflower alleviated liver oxidative injury and lipid peroxidation, via the upregulation of proteins such as 26S proteasome non-ATPase regulatory subunit 2 (PSMD2), cytochrome P450 2C6 (CYP2C6V1), and cytochrome P450 2C11 (CYP2C11), anddownregulation of arachidonate 12-lipoxygenase and 12S-type (ALOX12) protein expression. Conclusion: The water-soluble extract of safflower exhibited potential protective effects against lipid peroxidation and ethanol-driven oxidative damage within the liver via the upregulation of expression of proteins PSMD2, CYP2C6V1, and CYP2C11and downregulation of the expression of ALOX12, PDIA3, and TFG proteins.

Background: Polydatin has significant uricosuric effects. This study was conducted to examine the xanthine oxidase inhibition and to detect protein levels of urate transporter 1 (URAT1), organic anion transporters 1 (OAT1), and organic anion transporters 3 (OAT3) in the hyperuricemic mice to understand the role of antihyperuricemic mechanism of polydatin. Materials and Methods: Hyperuricemia mice model was established with uricase inhibitor (potassium oxonate), and uric acids in serum were observed. Kidney tissues were used to detect gene contents of URAT1, OAT1, and OAT3 by real-time-PCR (polymerase chain reaction) and to detect pathological features. The activity of xanthine oxidase (XOD) in the liver tissues of mice was detected. Results: The polydatin experimental groups and the positive control group (benzbromarone) showed significantly inhibited levels of serum uric acid when compared with the model group. Polydatin significantly inhibited the increasing tendency of the mRNA and the protein levels of OAT1 and OAT3, and decreased the tendency of the mRNA and the protein level of URAT1. Polydatin significantly inhibited the level of XOD in the liver tissues of mice in a concentration-dependent manner. Polydatin showed a protective effect on the pathological injury of the kidney. Conclusion: Polydatin has an antihyperuricemic effect in oxonate-induced hyperuricemic mice. The effect was related to the downregulation of renal URAT1, upregulation of renal OAT1 and OAT3, and inhibition of XOD in the hyperuricemic mice.

Background: Amygdalin is the essential component of the traditional Chinese medicinal drug peach kernel. The early studies of amygdalin are mainly focused on antitussive and tumor anti-tumor, but there are few studies on the treatment of neuroinflammation. Objectives: The purpose of this study was to explore the anti-inflammatory effect of amygdalin on lipopolysaccharide (LPS)-induced neuron–microglia and its protective mechanism for toll-like receptors (TLRs) signaling pathways. Materials and Methods: Griess, quantitative PCR, and flow cytometry were used to detect the influence of amygdalin on the release of inflammatory factors from BV2 mouse microglia (BV2 cells). The anti-inflammatory pathway of amygdalin was further explored by western blotting. At the same time, the effect of amygdalin on LPS-induced oxidative stress damage in BV2 cells was detected by WST-8 and fluorescence probe. Results: After LPS induction, the pro-inflammatory factors such as nitric oxide, interleukin-6, tumor necrosis factor-alpha, interleukin-1β, and recombinant NLR Family, Pyrin Domain Containing Protein3 were significantly increased. However, the levels were reversed significantly after treatment with amygdalin. In addition, amygdalin significantly inhibited the increase of TLR4, TLR2, and its downstream signal myeloid differentiation primary response gene 88 and P-NF-κB p65 in BV2 cells induced by LPS. Amygdalin can also reverse the abnormal expression of LPS-induced oxidative stress indexes such as cyclooxygenase-2, reactive oxygen species, and superoxide dismutase. Conclusion: Amygdalin may reduce the LPS-induced accumulation of pro-inflammatory substances via the down-regulating of the TLR2/TLR4-nuclear factor kappa-B signaling pathway.

Background: Allergic rhinitis (AR) is an allergic ailment of the immune-inflammatory system affecting people's daily lives. Scutellarin (SCU) is known for its inhibitory potential against GATA binding protein 3 (GATA3), an important mediator of AR. Aim: To determine the effect of SCU against AR induced by Ovalbumin (OVA) in the experimental animal. Materials and Methods: Mice (BALB/c strain) were induced with AR by sensitizing them with OVA (for consecutively 13 days) followed by the intranasal challenge (on day 21). They were simultaneously treated with either vehicle or montelukast (10 mg/kg) or SCU (50, 100, and 200 mg/kg). Results: Administration of SCU (100 and 200 mg/kg) markedly (P < 0.05) inhibited elevated nasal rubbing, sneezing, and nasal discharge, OVA-specific and, total immunoglobulin (Ig) E, IgG1 in serum, total and differential cell count, interleukins (ILs) levels in nasal lavage fluid. The up-regulated GATA3 and p-signal transducer and activator of transcription-6 (STAT6) expression of mRNA in the spleen were effectively (P < 0.05) reduced by SCU. Additionally, SCU effectively (P < 0.05) reduced OVA-induced alteration in nasal histopathology. Conclusion: SCU demonstrated an antiallergic effect against OVA-induced AR in experimental mice by inhibiting the GATA3/p-STAT6 pathway, thus improving the Th1/Th2 imbalance.

Background: Many studies have indicated that hair growth and color regulation are cyclical processes involved in the strict regulation of multiple signaling pathways, such as the transforming growth factor and Wnt/β-catenin protein pathways. It has also been reported that Glycyrrhiza uralensis extract can promote hair growth, but its active components are still unclear. Objectives: The aim of this study was to investigate the potential hair blackening effect and mechanisms of glycyrrhetic acid (GA) extracted from Glycyrrhiza uralensis. Materials and Methods: Different concentrations of GA solution (1.0 wt%, 10.0 wt%, and 20.0 wt%) were coated onto the skin surface of fur model mice induced by H₂O₂ for two weeks. After that, the number and depth of follicles and cortex thickness were observed in the skin tissue stained by hematoxylin and eosin (H&E). The relative mRNA levels of β-catenin, vascular endothelial growth factor (VEGF), and tyrosinase in skin tissue were analyzed using quantitative real-time polymerase chain reaction. The protein levels of β-catenin, VEGF, and tyrosinase were also evaluated by western blotting and immunohistochemistry. Results: There was a marked increase in the number of hair follicles and the blackening effect upon exposure to 10.0 wt% and 20.0 wt% GA. Moreover, the β-catenin, VEGF, and tyrosinase expression levels in the skin tissue varied significantly compared to those in the model group. Conclusion: GA showed positive effects on hair growth factor expression levels in mice, which meant that GA could promote hair blackening.

Background: Polygoni Multiflori Radix (PMR) is a widely distributed herb that has been used for centuries in the treatment of a range of systemic diseases among practitioners of traditional Chinese medicine. Objectives: The present comparative metabolite analysis study was conducted in an effort to understand the relationship between the geographical origin of PMR samples and their medicinal properties. Materials and Methods: As a metabolomics analysis of 35 PMR samples collected from Guangdong and other provinces in China was conducted via ultra-high performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry. Results: Differential metabolite profiles in these PMR samples were evaluated through multivariate statistical analyses. In total, this approach led to the identification of 778 differential metabolites [value of group contribution (VIP) >1, P < 0.01, and fold-change (FC) >2 or <0.5)] that were primarily associated with glycosaminoglycan degradation, lipoic acid metabolism, flavonoid biosynthesis, tyrosine metabolism, flavone and flavonol biosynthesis, phenylpropanoid biosynthesis, and phenylalanine metabolism. Of these metabolites, catechins accounted for seven significantly altered metabolites (VIP ≥2, P < 0.01, and FC >2 or <0.5) when comparing PMR samples from Guangdong with those from other regions. This suggests that PMR samples contain metabolite profiles characteristic of their provenance. Conclusion: As such, metabolomic profiling may be effective means of differentiating between PMR samples from different geographic regions within China, thus providing a sound theoretical basis for the reliable differentiation among and selection of pharmacologically optimal PMR samples.

Background: Breast cancer is the most common malignancy in women worldwide, so it is imperative to find new medications for this disease. The dry seeds of Brucea javanica have been extensively used in Traditional Chinese Medicine (TCM) for the treatment of tumor. Objectives: To isolate antitumor peptides from B. javanica seeds and research their antitumor activity with MCF-7 cells. Materials and Methods: Globulin of B. javanica was extracted and enzymatically hydrolyzed by pepsin. Then ultrafiltration, gel filtration chromatography (GFC), and reverse-phase high-performance liquid chromatography (RP-HPLC) were employed to purify the peptides. Anticancer activity was studied in humans in human breast cancer MCF-7 cells, MTT (3- [4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide)-assessed cell viability, Giemmsa staining determined whether the cells should undergo apoptosis, and apoptosis and cycle assays assessed the antitumor mechanism of the cells. Real time quantitative polymerase chain reaction (RT-qPCR) was used to study the alterations of oncogenes and cancer metastasis suppressor genes in MCF-7 cells treated with peptide extracts. Results: The fraction F9-9 extracted from B. javanica significantly inhibited the proliferation of MCF-7 cells with the IC₅₀ of 0.124 ± 0.004 μg/mL. The possible mechanisms of this fraction were further explored. F9-9 obviously induced cell cycle arrest at G0/G1 phase and promoted apoptosis of MCF-7 cells. The underlying molecular mechanisms were explored by RT-qPCR and results showed that in B. javanica small molecular peptides upregulated the expression of tumor suppressor genes P53 and PTEN as well as tumor metastasis suppressor gene NM23H-1. Conclusion: This study suggests that B. javanica peptides have therapeutic activity and mechanisms for antibreast cancer, encouraging further isolation and purification to obtain monomeric peptides of this extract for anticancer use.

Background: The number of diabetic patients worldwide continues to increase. Diabetes mellitus (DM) has become one of the three chronic diseases threatening human health. Mangiferin is a kind of xanthone with multiple biological activities. Studies have confirmed that it has good activity in treating DM. Objectives: The purpose of our study was to investigate the targets and mechanism of mangiferin in the treatment of DM and to analyze its metabolites in vivo. Materials and Methods: We predict the targets and mechanism of mangiferin in DM in view of network pharmacology (NP). Mangiferin's targets were completed using Gene Cards, Swiss Target Prediction, and TCMSP database. DisGeNET database retrieved DM-related targets. The common targets were put into STRING platform to construct a protein–protein interaction (PPI) network model. DAVID platform and Cytoscape were used to achieve GO analysis and KEGG signal pathway enrichment analysis. Then, in metabolites study, LC-MS/MS was used to analyze the metabolites of mangiferin in plasma collected from SD rats. Results: A total of 37 targets for the coaction of mangiferin and DM were screened. Tumor necrosis factor (TNF), epidermal growth factor (EGF), prostaglandin-endoperoxide synthase 2 (PTGS2), estrogen receptor 1 (ESR1), hypoxia-inducible factor-1 alpha (HIF1A), nuclear factor-κB p65 (RELA), protein kinase C alpha (PRKCA), and Interleukin-2 (IL2) were selected as the most important targets. The biological functions related to DM were mainly enriched in the signaling pathway in phosphoinositide 3-kinase (PI3K)-protein kinase B (Akt), mitogen-activated protein kinase (MAPK), nuclear factor-kappa B (NF-kappa B), hypoxia inducible factor-1 (HiF-1), and mammalian target of rapamycin (mTOR). With reference to the obtained accurate relative molecular mass, chromatographic retention behavior, and characteristic fragment ions, a total of six metabolites including the original drug were analyzed and identified. Conclusion: This study provides key data for the mechanism of action and targets research on mangiferin, and it provides a basis for its further development into hypoglycemic drugs.

Background: Lactuside B (LB) is a novel active ingredient separated from Pterocypsela elata by our team. In our preliminary study, LB showed a therapeutic effect on depression. However, the underlying molecular mechanisms remain unclear. Objectives: This study was the first to study the pharmacological effects and mechanisms of LB on depression. Materials and Methods: Network pharmacology method was applied to screen the candidate targets and signaling pathways. Furthermore, molecular docking, cell test, and animal experiments were used to confirm the antidepression mechanisms. Results: The network pharmacology results showed 29 targets and 40 signaling pathways of LB on depression. Molecular docking showed that there was a strong binding effect between LB and each target (cAMP response element binding protein B [CREB], Ras, Raf, and ERK1/2). Cell tests indicated that the expressions of Ras and CREB in astrocyte cells (0.05–0.20 g/L of LB) exhibited significant differences (P < 0.05) compared with the model group, respectively. In animal tests, the expressions of Ras, ERK1/2, and Raf (12.5–50.0 μg/10 g of LB) showed a significant difference compared with mice in the model group (P < 0.05). Conclusion: These results indicated that the antidepression mechanism of LB was mainly associated with Ras signaling pathway, and the cAMP signaling pathway and PI3K-Akt signaling pathway may play an essential role.

Background and Aim: Intestinal ischemia/reperfusion (II/R) can cause injury of remote organs, including acute renal injury. All of these, in turn, increase the rate of disability and mortality. Anwulignan (Anwu) is an active monomer from Schisandra sphenanthera, which has been used for thousands of years in China as a medical herb. In our previous study, we found that Anwu can improve the intestinal function after ischemia/reperfusion. Therefore, we were curious to know if Anwu can have a protective effect on remote organ injury after II/R. The purpose of the present study was to examine the effect of Anwu on the remote renal injury induced by II/R in rats and investigate its mechanism. Materials and Methods: Forty Sprague Dawley (SD) rats were divided into four groups, and they were sham operation (Sham), sham + Anwu, II/R, and II/R + Anwu groups. After reperfusion or sham operation, blood and kidneys were collected from the rats for the detection of relative biochemical parameters. Results: Renal indexes were not significantly different among all groups. In the II/R group the following were the findings: blood urea nitrogen and creatinine levels were increased, kidney injury score was increased, renal superoxide dismutase was reduced, the activities of reduced glutathione and catalase were decreased, the renal malondialdehyde content was increased, there was an improvement in the levels of renal proinflammatory cytokines, interleukin-6, tumor necrosis factor-α, and interleukin-1β, the expressions of renal oxidative stress-related p-Nrf2 and heme oxygenase (decycling) 1 (HO-1) were decreased, and the Kelch-like ECH-associated protein 1 (Keap1) expression was increased. The expression levels of the apoptotic protein, cleaved caspase-3, were raised, while the B-cell lymphoma 2 (Bcl-2)/Bcl-2-associated x protein (Bax) ratio was reduced. However, the administration of Anwu before the surgery significantly reversed the above changes. Conclusion: Anwu has a protective effect against the II/R-induced remote renal injury in rats, which may be related to its regulation of antioxidant, anti-inflammatory, and antiapoptotic pathways.

Background: Radix Bupleuri-Radix Paeoniae Alba (CH-BS) is a common clinical herb pair, which is often used to treat depression. However, there is little known about the difference in serum chemical profile before and after the compatibility of Radix Bupleuri (CH) and Radix Paeoniae Alba (BS). Objectives: Here, an effective strategy has been developed for screening differences of constituents in vivo before and after compatibility by integrating background subtraction and metabolomics technology. Materials and Methods: First, to obtain the chemical composition from the CH in vivo before compatibility, the Liquid chromatography/Mass spectrometry (LC/MS) data of serum samples in the control group were subtracted from serum samples in the CH group based on the background subtraction method. Moreover, the chemical composition from the CH in vivo after compatibility was obtained by subtracting the LC/MS data of serum samples in the BS group from serum samples in the CH-BS compatibility group. Finally, the difference in the chemical composition from the CH in vivo before and after compatibility was screened and analyzed by metabolomics. Meanwhile, the difference in chemical composition from the BS in vivo before and after the compatibility was found according to the same thought with BS. Results: The results showed that three prototype components were significantly decreased and seven metabolites were significantly increased in CH after compatibility. And eight prototype components and six metabolites were significantly increased in BS after compatibility. Conclusion: An effective strategy has been developed for screening differences of constituents in vivo before and after compatibility by combining background subtraction and metabolomics.

Background: Shikonin (SKN) is a widely used Chinese Traditional Medicine with anti-inflammatory and anti-arthritic properties. Objectives: The research aimed to study the anti-cancer effect of SKN in diethylnitrosamine (DEN) and ferric nitrilotriacetate (Fe-NTA)-induced renal carcinoma in rats. Materials and Methods: The experimental animal groups consisted of Group I (Control), Group II (DEN + Fe-NTA), Group III (DEN + SKN), and Group IV (SKN alone). Retroperitoneal infusions were given for 24 weeks. Subsequently, tissues and blood samples were tested for biochemical, histopathologic, enzyme-linked immunosorbent assay, and western blot tests. Results: After 24 weeks, the antioxidant enzymes were found to increase in the SKN-treated groups. Histopathology revealed normal tissue morphology with a significantly reduced inflammatory response in SKN-treated animal groups compared to the control group. The serum levels of necrosis factor kappa B, PGE₂, interleukin-1β, interleukin-6, and tumor necrosis factor-alpha were also down-regulated in the SKN-treated animal groups. The apoptotic proteins (Caspase-3, -9, and Bcl-2-associated X protein) were higher confirming SKN-induced apoptosis. Conclusion: Overall evidence suggests that SKN exhibits a reno-protective anti-cancer effect against DEN + Fe-NTA induced renal carcinoma in rats. Thus, SKN emerges as a therapeutic agent for kidney cancer therapy.

Background: There is an increasing interest in new natural compounds for the treatment of myocardial infarction (MI). Gedunin is a tetranortriterpenoid extracted from the Indian neem tree (Azadirachta indica). The main objective of this study is to understand the cardioprotective effects of gedunin against MI through the suppression of NF-κB-mediated inflammatory pathways. Materials and Methods: Male Wistar rats were categorized into 4 groups of 6 each: control, inducer isoproterenol (ISO) alone, ISO + gedunin-treated, and gedunin (50 mg/kg b.w.)-pretreated rats followed by ISO induction. The infarct size, heart-to-body weight ratio, cardiac enzymes, ATP values, Ca²⁺ levels, and inflammatory and apoptotic markers were studied and compared in all these groups. Data were expressed as mean ± SD. One-way analysis of variance (ANOVA) and post hoc Tukey–Kramer test were used for comparing multiple values. Results: Gedunin pre-treatment caused a reduction in cardiac size, attenuated the levels of cardiac bio-enzymes, and reduced immune cell infiltration and necrosis. It also enhanced the secretion of antioxidant enzymes glutathione-S-transferase (GST) and superoxide dismutase (SOD) and reduced glutathione (GSH) and glutathione peroxidase (GPx). Gedunin suppressed inflammation by down-regulating the secretion of interleukin 10 (IL-10), interleukin-1β (IL-1β), nuclear factor-kappa B (NF-κB), tumor necrosis factor-α (TNF-α), and apoptotic markers caspase-3, caspase-9, Bax, and Bcl-2. Conclusion: Gedunin inhibited apoptosis in cardiotoxicity-induced MI in rats by inhibiting inflammation and apoptosis markers. Thus, gedunin is a potential natural cardioprotective compound.

Aim: Thyroid cancer accounts for 3.8% of the total endocrine cancer and is the fifth most diagnosed cancer among Indian women. Geraniol possesses pharmacological properties, including anti-inflammatory, anti-cancer, and neuroprotective properties. The anti-cancer activity of geraniol on TPC-1 cells was assessed in this study. Materials and Methods: The anticancer activity of geraniol in TPC-1 cells was evaluated by MTT test, ROS, DAPI, PI, and AO/EB staining. mRNA and expression were assessed by qPCR and western blot. Results: The geraniol has shown cytotoxic effects toward TPC-1 cells and showed an IC₅₀ value of 25 μM, which decreases cell viability and inhibited cell proliferation. It also increased the accumulation of intracellular ROS, which induces apoptosis in the TPC-1 cells due to the dissipation of membrane potential and disruption of mitochondria. There is a substantial reduction in the mRNA level expressions of Bcl-2, Cyclin D1, c-Myc, COX-2, TNF-α, NF-κB, IL-6, and surviving. It also upregulated the Bax and caspase-3 mRNA expression suggesting cell death-mediated apoptosis. Conclusion: We report that geraniol effectively inhibited the JAK-2, STAT-3, and ERK signaling pathways depicting geraniol as a potential therapeutic molecule for thyroid cancer treatment.

Background: Withaferin A is a triterpenoid steroidal lactone and is commonly isolated from Withania somnifera. It has been shown to have tremendous pharmacological potential. Objectives: The present study was aimed at elucidating the mechanism of action of withaferin A against hepatocellular carcinoma (HCC) cells via the PI3K/AKT signaling pathway. Materials and Methods: MTT (tetrazolium dye) Assay was performed for the relative assessment of the viabilities of cell lines. The effect of withaferin A on the proliferative capability of HCC cells was analyzed using the EdU staining assay, and the colony-forming potential was assessed with the help of a clonogenic assay. Cell apoptosis was estimated through (Modified Annexin V/Propidium Iodide Apoptosis Assay) staining followed by flow cytometry. Transwell assays were carried out for estimating the migration and invasion of cancer cells. The qRT-PCR and western blotting, respectively, were used for gene and protein expression studies. Results: Withaferin A selectively inhibited the viability of HCC cells although the viability of normal liver cells was minimally affected. The IC₅₀ of withaferin A against the HepG2 cells was found to be 12 μM as against 150 μM for normal THLE-2 cells. The treatment with withaferin A significantly minimized the proliferation and colony-forming potential of cancer cells by inducing cell apoptosis. The percentage of apoptosis increased from 3.8% in control to 20.3% at 24 μM withaferin A. The cancer cell migration and invasion were significantly declined by withaferin A together with the inhibition of Epithelial to mesenchymal transition (EMT) of HCC cells. The withaferin A treatment decreased the expression of carcinoma cell markers CD44, CD90, and EpCAM. The expression of miR-200c markedly increased under withaferin A treatment and the latter was shown to exert its anti-cancer effects through miR-200c-mediated inhibition of the PI3K/AKT signaling pathway in HCC cells. Conclusion: In conclusion, withaferin A modulated the expression of miR-200c in HCC cells to inhibit the PI3K/AKT pathway and restricted the cancer cell EMT together with the inhibition of in vitro cancer cell growth and viability via induction of apoptosis.

Background: Rabdosia rubescens is a traditional herbal medicine, commonly known as an anti-cancer plant. A large number of literatures have reported that the diterpenoids in R. rubescens have strong anti-tumor effects. The purpose of this study was to isolate the anti-cancer active ingredients from R rubescens, identify their structures by nuclear magnetic resonance (NMR), and establish high-performance liquid chromatography (HPLC) method for the determination of content of the oridonin and the ponicidin in R. rubescens. Materials and Methods: The anti-cancer components of R. rubescens were isolated using silica gel column chromatography, and their structures were identified by NMR technology. HPLC method was established for the determination of the content of the oridonin and the ponicidin in R. rubescens collected from July to October in Taihang Mountain, China. Results: Two anti-cancer components—the oridonin and the ponicidin—were isolated and identified. HPLC methods for the determination of the oridonin and the ponicidin were established. This method could be used to complete the chromatographic analysis of the oridonin and the ponicidin within 10 min, and the chromatographic peaks of each component reached the baseline separation. The contents of the oridonin and the ponicidin in the whole grass of R. rubescens collected from July to October were 0.469%, 0.618%, 0.625%, 0.448% and 0.124%, 0.203%, 0.216%, 0.127%, respectively. The results showed that August to September was the best harvest time, and the contents of the oridonin and the ponicidin were the highest during this time. The specific harvest time was determined by the climate and precipitation of the year. Conclusion: The HPLC method established in this study was simple, rapid, accurate, reliable, and reproducible, which provided a reference for drug application and resource utilization of R. rubescens and could be used for quantitative analysis of anti-cancer active ingredients in R. rubescens.

Background: Overexpression of transforming growth factor-β (TGF-β) and its cancer regulators are the major phenomena in oral cancer. The study aimed to investigate the anticancer effect of degalactotigonin (DGT), a steroidal glycoside from Solanum nigrum on oral cancer. Objectives: The study aimed to investigate the anti-invasion and apoptotic induction effect of DGT in oral squamous cell carcinoma (OSCC) cells through inhibiting non-canonical TGF-β signalling. Materials and Methods: Human oral cancer KB (KERATIN-forming tumor cell line HeLa) cells were chosen to study the anti-cancer activity of DGT in vitro experiments. The cytotoxic effect of DGT was evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) assay. Cell growth, DNA damage, invasion inhibition and apoptosis activation were evaluated by mitochondrial membrane potential (ΔΨM), comet assay, reactive oxygen species (ROS) and AO/EtBr staining. The DGT effect on protein expression levels related to invasion, apoptotic markers and its activation signalling pathways on KB cells were examined by western blotting. Results: We found that DGT antioxidant properties reduced cell viability, generates ROS, enhanced DNA damage, and MMP dissipation. Further, TGF-β inhibitions resulted in a reduction of extracellular signal-regulated kinase (ERK), NF-κB and activation of JNK, p38 which increase ROS in a cancer cell that downregulates cyclin-D1, PCNA, MMP-2, MMP-9, Bcl-2 and increases Bax, Caspase-9, Caspase-3 protein expressions that simultaneously subdues the cancer developments. Conclusion: These findings suggest that DGT inhibiting TGF-β mediated ERK, NF-κB and activation of JNK, p38 caused tumour cell death via ROS stimulation and apoptosis.

Background: Ulcerative colitis (UC) is a prevalent case which needs more detailed exploration. We attempted to illustrate how miR-223-3p acted in the development of UC. Materials and Methods: The level of miR-223-3p in serum from UC patients was detected using quantitative reverse-transcriptase polymerase chain reaction. The in vivo experimental spleen-kidney-yang deficiency mice model was established to get a better understanding of the action of miR-223-3p in UC development. Hematoxylin and eosin staining was applied to observe the intestinal epithelial barrier function. Western blotting was applied to measure the expression of ZO-1, Occludin, TGF-β, and Smad3. Results: Enrichment of miR-223-3p was observed in serum from UC patients. miR-223-3p antagomir could lower the shortening of colon length, alleviating the intestinal epithelial barrier function damage caused by UC. Under the UC conditions, TNF-α, IL-6, and IL-1β were up-regulated, whereas with the miR-223-3p antagomir, the up-regulation was less pronounced; ZO-1 and Occludin protein levels were decreased, whereas the decrease was less obvious in the miR-223-3p antagomir group. Additionally, miR-223-3p regulated UC development through the TGF-β/Smad3 pathway. Conclusion: miR-223-3p promoted UC development with spleen-kidney-yang deficiency through the TGF-β/Smad3 pathway.