Background: Plant-based natural extracts cure several diseases in human. However, the extract of Psidium guajava leaf is not yet evaluated on changes of lipid profile in hepatic disease affected rats. Objective: The present study was aimed to evaluate the mitigation effect of the ethanolic extract of P. guajava leaf and its isolated quercetin fraction on hepatotoxic rats. Materials and Methods: Carbon tetrachloride (CCl₄) was injected to rats for hepatic disease induction and silymarin drug was used as positive control to compare plant ethanolic extract. The lipid profiles were assessed in both plasma and liver tissue of diseased and control rats. Results: Levels of total cholesterol, triglycerides, free fatty acids, phospholipids, and low-density lipoprotein cholesterol were increased and the level of high-density lipoprotein cholesterol (HDL-C) was decreased in CCl₄-induced hepatotoxic rats. The treatment of P. guajava (100, 200, and 300 mg/kg, bw) and isolated quercetin fraction (20 mg/kg, bw) doses decreased the elevated levels of all these parameters in diseased rats and restored the normal concentration of HDL-C. Conclusion: The results of the present study concluded that the P. guajava leaf and its isolated quercetin fraction can significantly regulate lipid metabolism in CCl₄-induced hepatotoxic rats and decrease the disease rate. Abbreviations used: CCl₄: Carbon tetrachloride; FFA: Free fatty acids; HDL-C: High-density lipoprotein cholesterol; LCAT: Lecithin cholesterol acyltransferase; LDL-C: Low-density lipoprotein cholesterol; PL: Phospholipids; TC: Total cholesterol; TG: Triglycerides; VLDL-C: Very low-density lipoprotein cholesterol.

Background: Parkinson's disease (PD) is a progressive neurodegenerative disorder (NDD) associated with the loss of dopaminergic neurons in the substantia nigra and subsequently has an effect on motor function and coordination. The pathology of PD is multifactorial, in which neuroinflammation and oxidative damage are the two of the main protagonists. Objectives: The present study aims to assess the potential antioxidant and anti-inflammatory effects of demethoxycurcumin (DMC), a natural derivative of curcumin, against rotenone-induced PD in rats. Materials and Methods: Rats were randomized and divided into six groups: control, rotenone (0.5 mg/kg/day, intraperitoneal in sunflower oil) treated for 7 days, rotenone and DMC (5, 10, and 20 mg/kg b.w) cotreated, and DMC (20 mg/kg b.w) alone treated groups. Results: Based on the dopamine concentration and biochemical estimations, the effective dose of DMC was selected and the chronic study was performed. At the end of the experimental period, behavioral studies and protein expression patterns of inflammatory markers were analyzed. Rotenone treatment led to motor dysfunctions, neurochemical deficits, and oxidative stress and enhanced expressions of inflammatory markers, whereas oral administration of DMC attenuated all the above. Conclusion: Even though further research is needed to prove its efficacy in clinical trial, the results of our study showed that DMC may offer a promising and new therapeutic lead for the treatment of NDDs including PD. Abbreviations used: COX-2: Cyclooxygenase-2; DA: Dopamine; DMC: Demethoxycurcumin; DMRT: Duncan's multiple range test; GSH: Reduced glutathione; GPx: Glutathione peroxidase; IL-1 β: Interleukin-1 β; IL-6: Interleukin-6; iNOS: Inducible nitric oxide synthase; PD: Parkinson's disease; SN: Substantia nigra; SOD: Superoxide dismutase; TBARS: Thiobarbituric acid reactive substances; TNF-α: Tumor necrosis factor-α.

Background: “Decocting later” is important procedure for the extraction of herbal medicines containing volatile compounds. Objective: This study was performed to investigate optimal conditions for “Decocting later” of Menthae herba in Eungyo-san (EGS) and correlation between extraction variables and the yields of d/l-menthol, a marker compound of Menthae herba. Materials and Methods: The decocting temperature, total decocting time, and decocting later time were chosen as individual variables, and the yield of d/l-menthol was set as the response value which were calculated by using a Box-Behnken design (BBD). The amount of d/l-menthol was quantified using gas chromatography/mass spectrometry. Results: Response surface methodology (RSM) was used to predict optimal conditions for decocting later of Menthae herba into the formula. Optimal conditions for “Decocting later” from RSM were as follows: 100.63°C of decocting temperature; 82.95 min of total decocting time; 19.11 min of decocting later time. Both decocting temperature and total decocting time showed significant correlation with the yield of d/l-menthol. Conclusions: These results suggest that the decocting temperature and total decocting time were influential factors, and RSM can be applied for optimizing the conditions of “Decocting later” of Menthae herba in EGS. Abbreviations used: KM: Korean medicine; EGS: Eungyo-san; GC/MS: Gas chromatography/mass spectrometry; RSM: Response surface methodology; SIM: Selected ion monitoring; LOD: Limits of determination; LOQ: Limits of quantification; RSD: Relative standard deviation; ANOVA: Analysis of variance; BBD: Box-Behnken design.

Background: Silybum marianum, a member of the Aster family, is a well-known Chinese herb and the source of a popular antioxidant that is extensively used in Asia. The abundant S. marianum straws are still underutilized and wastefully discarded to pollute the environment. Objective: To solve the above problem and better utilize S. marianum straws, the objective of this study was to optimize the conditions for extraction of silymarin from S. marianum straws. Materials and Methods: A combination of microwave-assisted extraction and response surface methodology (RSM) was used for silymarin from S. marianum straws and yield assessment by high-performance liquid chromatography method. The RSM was based on a five-level, four-variable central composite design (CCD). Results: The results indicated that the optimal conditions to obtain highest yields of silymarin were microwave power of 146 W, extraction time of 117 s, liquid-to-solid ratio of 16:1 mL/g, and ethanol concentration of 43% (v/v). Validation tests indicated that under the optimized conditions, the actual yield of silymarin was 6.83 ± 0.57 mg/g with relative standard deviation of 0.92% (n = 5), which was in good agreement with the predicted yield. Conclusions: The exploitation of the natural plant resources present very important impact for the economic development. The knowledge obtained from this work should be useful to further exploit and apply this material. Abbreviations used: MAE: Microwave-assisted extraction, RSM: Response surface methodology, HPLC: High-performance liquid chromatography, CCD: Central composite design, ANOVA: Analysis of variance.

Background: Camel milk (CM) and Nigella sativa (NS) have been traditionally claimed to cure wide range of diseases and used as medicine in different part of world, particularly in Saudi Arabia. Several research studies have been published that proved beneficial effects of CM and NS. Objective: This study was undertaken to investigate the antihepatotxic potential of CM and NS oil (NSO) against thioacetamide (TAA)-induced hepato and nephrotoxicity in rats. Materials and Methods: Thirty female Albino Wistar rats were randomly divided in to six groups having five rats in each group. A single subcutaneous injection of TAA (100 mg/kg b. w.) was administered to all the rats in Group-II to VI on 1^st day to induce hepatorenal damage. Group I served as a normal control while Group II served as toxic control for comparison purpose. Experimental animals in Group III, IV, and V were supplemented with fresh CM, (250 mL/24 h/cage), NSO (2 mL/kg/day p. o.), and NSO + fresh CM, respectively. Group VI was treated with a polyherbal hepatoprotective Unani medicine Jigreen (2 mL/kg/day p. o.) for 21 days. TAA-induced hepatorenal damage and protective effects of CM and NSO were assessed by analyzing liver and kidney function tests in the serum. Histopathology of liver and kidney tissues was also carried out to corroborate the findings of biochemical investigation. Results: The results indicated that the TAA intoxicated rats showed significant increase in the alanine transaminase, aspartate transaminase, gamma-glutamyl transpeptidase, alkaline phosphatase, lipid profile, urea, creatinine, uric acid, sodium, and potassium levels in serum. Treatment of rats with CM, NSO, and CM plus NSO combination and Jigreen significantly reversed the damage and brought down the serum biochemical parameters and lipid profile toward the normal levels. The histopathological studies also support the hepato and nephroprotective effects of CM and NSO. Conclusion: This study demonstrated the ameliorative effects of CM, NSO, and CM plus NSO combination against TAA-induced hepatorenal toxicity in rats. Abbreviations used: CM: Camel milk; NS: Nigella sativa; NSO: Nigella sativa Oil; TAA: Thioacetamide; S.C.: Subcutaneous; Jig: Jigreen; b.w.: Body Weight; mL: Milli liter; mg: Milli gram; g: Gram; Kg: Kilo gram; ALT: Alanine transaminase; AST: Aspartate transaminase; GGT: Gamma-Glutamyl Transpeptidase; ALP: Alkaline Phosphatase; TC: Total Cholesterol; HDL-C: High Density Lipoprotein Cholesterol; LDL-C: Low Density Lipoprotein Cholesterol; TG: Triglyceride; TB: Total bilirubin; K⁺: Potassium; Na⁺: Sodium; CCl₄: Carbon Tetrachloride; °C: Degree Celsius; p.o.: Per Oral; RPM: Revolutions per minute; H&E: Hematoxylin and Eosin; SEM: Standard Error of Mean; ANOVA: The one-way analysis of variance.

Background: Crocus sativus L. (saffron) is a scarce plant that has been used as food flavoring agent, coloring agent, and traditional herbal medicine. Methods: The bioactivity of exopolysaccharide (EPS) extracted from an endophytic fungus of C. sativus was examined for the first time by antioxidative, antitumor, and antibacterial assays. The extraction conditions for EPS were optimized by combining the response surface methodology with Box-Behnken design. Results: EPS exhibited excellent scavenging activities against 1,1-diphenyl-2-picrylhydrazyl, hydroxyl and superoxide anion radicals, and moderate cytotoxicities against K562, A549, HL-60, and HeLa cells. The optimum extraction conditions for EPS were as follows: precipitation time of 16 h, precipitation temperature of 3.7°C, pH 7.2, and ratio of ethanol to fermented broth of 5:1 (L/L). Under the optimized conditions, the yield of EPS reached 162 ± 6 μg/L which was close to the predicted one (165 μg/L). Moreover, high-performance liquid chromatography of monosaccharide composition showed that EPS comprised mannose, glucose, galactose xylose, and arabinose in a molar ratio of 25.6:16.5:1.0:3.8:5.4. Conclusion: EPS may be an eligible substitute for C. sativus and a potential bioactive source applicable to pharmaceutical and food industries. Abbreviations used: EPS: Exopolysaccharide, RSM: Response surface methodology, BBD: Box-Behnken design, DPPH: 1,1-diphenyl-2-picrylhydrazyl, V_C: Ascorbic acid, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, LB: Luria Bertani, DMSO: dimethyl sulfoxide, PMP: 1-phenyl-3-methyl-5-pyrazolone, FT-IR: Fourier transform-infrared, HPLC: High-performance liquid chromatography, 3D: Three-dimensional, 2D: Two-Dimensional.

Background: Hypertension is a great global health challenge and it mostly requires drug combination therapy with the various advantages. Metoprolol (MP) and paeoniflorin are both commonly used for the treatment of hypertension. However, whether they exert synergistic effects on antihypertension or not remains unclear, especially on vascular endothelial function. Objective: The purpose of the study is to investigate the advantages of the combined antihypertensive effects of paeoniflorin enriched extract from Radix Paeoniae Alba (RE) and MP in spontaneously hypertensive rats (SHR). Materials and Methods: SHR divided into six groups (n = 8 each group), animals in each group were administrated orally with distilled water, MP (6 and 20 mg/kg), RE (30 and 90 mg/kg), and MP (6 mg/kg) combined with RE (30 mg/kg) (MP + RE), respectively, daily for 6 weeks. Blood pressure (BP) and microcirculation were assessed. The organ bath experiment and hematoxylin and eosin staining were, respectively, performed for the functional and pathological vascular function analysis. Immunohistochemistry was applied to detect endothelial nitric oxide synthase (eNOS) expression in aorta, heart, and kidney. Further, high-performance liquid chromatography was employed to quantitatively determine paeoniflorin in RE and MP + RE sample solvent, as well as in plasma of Sprague-Dawley rats (SD) after single-dose administration of them. Results: The results showed that MP + RE significantly reduced BP, increased microcirculation, improved vascular function and pathological changes, and upregulated eNOS expression. MP was also found to increase the blood concentration of paeoniflorin in SD. Conclusion: The combination of RE and MP could be used for the treatment of hypertension and could improve microcirculation, upregulate eNOS expression, and mitigate endothelial dysfunction in SHR. Abbreviations used: RE: Paeoniflorin enriched extract from Radix Paeoniae Alba, MP: Metoprolol, MP + RE: MP combined with RE, NC: Normal control, MC: Model control, SHR: Spontaneously hypertensive rats, SD: Sprague-Dawley rats, H and E: Hematoxylin and eosin, BP: Blood pressure, SBP: Systolic blood pressure, DBP: Diastolic blood pressure, MBP: Mean arterial blood pressure, NA: Norepinephrine, ACh: Acetylcholine, SNP: Nitroprusside, NO: Nitric oxide, eNOS: Endothelial nitric oxide synthase, RPA: Radices Paeoniae Alba, IHC: Immunohistochemistry, C_max: Peak concentration, T_max: The time to reach C_max, t_½: Half-life, AUC_0-t: Area under the curve of 0-t time; MRT_0-t: Mean residence of 0-t time; CL: Clearance rate.

Introduction: Aerva lanata (Linn) of family Amaranthaceae is an important and commonly used plant for its medicinal and pharmacological properties and proving the traditional uses of flowers of A. lanata Linn. Objective: All extracts of A. lanata were further evaluated for antioxidant, antimicrobial, and antiurolithiatic potential to scientifically prove the traditional uses. Materials and Methods: In the present investigation, different solvent extracts of flowers were obtained using a Soxhlet extractor. Microorganisms were obtained from IMTECH, Chandigarh. Antiurolithiatic study was carried out in Albino Research and Training Centre, Hyderabad. Results: Regardless of the antioxidant studied, the methanolic extract presented the highest antioxidant activity and the aqueous extracts offered the lowest, following the order: methanolic extract > ethyl acetate > chloroform > aqueous. The results of this antimicrobial study indicate that methanolic extract of A. lanata could be used as antimicrobial agents. Overall, the methanolic flower extract of A. lanata (Linn) was significantly more promising as antiurolithiatic spectrum. This result also suggested the potential usefulness of the methanolic extract as an antiurolithiatic agent. Conclusion: Henceforward, this research can be acknowledged as a prime new report that focuses on the application of A. lanata (Linn) as an antioxidant, antimicrobial, and antiurolithiatic agent. Abbreviations used: IMTECH Chandigarh: Institute of Microbial Technology, Chandigarh; IMMT: Institute of Mineral and Material Technology; CSIR: Council of Scientific & Industrial Research; DPPH: 1,1-diphenyl-2-picrylhydrazyl; MTCC: Microbial Type Culture Collection; BHT: Butylated Hydroxyl Toluene.

Background: In traditional folk medicine, Limonium tetragonum is used in the treatment of uterine hemorrhage, tinnitus, and oligomenorrhea. Objective: This study aimed to identify the therapeutic effect of L. tetragonum EtOAc extract (EALT) on liver of mice with chronic alcohol poisoning. Materials and Methods: C57BL/6J mice were administered 100 mg/kg of EALT with a single binge ethanol/Lieber-DeCarli liquid diet for 8 weeks. Results: The chronic-binge ethanol diet induced a significant increase in liver marker enzyme activities. Coadministration of EALT reversed the elevation of serum total cholesterol and triglyceride as well as aspartate aminotransferase and alanine aminotransferase due to chronic alcohol consumption. Histologic findings including markedly attenuated fat accumulation in hepatocytes were observed in EALT-treated mice. EALT supplementation prevented alcoholic liver injury through attenuation of inflammatory mediators such as toll-like receptor-4, cytochrome P4502E1, and cyclooxygenase-2, and inflammatory cytokine interleukin-6. Conclusion: Results provided direct experimental evidence for the hepatoprotective effect of EALT in the NIAAA mouse model. Therapeutic attempts with the L. tetragonum extract might be useful in the management of alcoholic liver disease. Abbreviations used: EALT: L. tetragonum EtOAc extract; TC: Total cholesterol; TG: Triglyceride; ROS: Reactive oxygen species; CYP2E1: Cytochrome P4502E1; TLR-4: Toll-like receptor-4; COX-2: Cyclooxygenase-2.

Background: Citri Reticulatae Pericarpium is the dried mature pericarp of Citrus reticulata Blanco which can be divided into “Chenpi” and “Guangchenpi.” “Guangchenpi” is the genuine Chinese medicinal material in Xinhui, Guangdong province; based on the greatest quality and least amount, it is most expensive among others. Hesperidin is used as the marker to identify Citri Reticulatae Pericarpium described in the Chinese Pharmacopoeia 2010. However, both “Chenpi” and “Guangchenpi” contain hesperidin so that it is impossible to differentiate them by measuring hesperidin. Objective: Our study aims to develop an efficient and accurate method to separate and identify “Guangchenpi” from other Citri Reticulatae Pericarpium. Materials and Methods: The genomic deoxyribonucleic acid (DNA) of all the materials was extracted and then the internal transcribed spacer 2 was amplified, sequenced, aligned, and analyzed. The secondary structures were created in terms of the database and website established by Jörg Schultz et al. High-performance liquid chromatography-diode array detection-electrospray Ionization/mass spectrometry (HPLC-DAD-ESI-MS)/MS coupled with chemometric analysis was applied to compare the differences in chemical profiles of the three kinds of Citri Reticulatae Pericarpium. Results: A total of 22 samples were classified into three groups. The results of DNA barcoding were in accordance with principal component analysis and hierarchical cluster analysis. Eight compounds were deduced from HPLC-DAD-ESI-MS/MS. Conclusions: This method is a reliable and effective tool to differentiate the three Citri Reticulatae Pericarpium. Abbreviations used: CTAB: Hexadecyltrimethylammonium bromide, DNA: Deoxyribonucleic acid, ITS2: Internal transcribed spacer 2, PCR: Polymerase chain reaction.

Background: Noni juice has been extensively used as folk medicine for the treatment of arthritis, infections, analgesic, colds, cancers, and diabetes by Polynesians for many years. Due to the lack of standard scientific evaluation methods, various kinds of commercial Noni juice with different quality and price were available on the market. Objective: To establish a sensitive, reliable, and accurate high-performance liquid chromatography with electrospray ionization triple quadrupole mass spectrometry (HPLC-ESI-MS/MS) method for separation, identification, and simultaneous quantitative analysis of bioactive constituents in Noni juice. Materials and Methods: The analytes and eight batches of commercially available samples from different origins were separated and analyzed by the HPLC-ESI-MS/MS method on an Agilent ZORBAX SB-C₁₈ (150 mm × 4.6 mm i.d., 5 μm) column using a gradient elution of acetonitrile-methanol-0.05% glacial acetic acid in water (v/v) at a constant flow rate of 0.5 mL/min. Results: Seven components were identification and all of the assay parameters were within the required limits. Components were within the correlation coefficient values (R² ≥ 0.9993) at the concentration ranges tested. The precision of the assay method was <0.91% and the repeatability between 1.36% and 3.31%. The accuracy varied from 96.40% to 103.02% and the relative standard deviations of stability were <3.91%. Samples from the same origin showed similar content while different origins showed significant different result. Conclusions: The developed methods would provide a reliable basis and be useful in the establishment of a rational quality control standard of Noni juice. Abbreviations used: HPLC-ESI-MS/MS: High-performance liquid chromatography with electrospray ionization triple quadrupole mass spectrometry, LOD: Limit of detection, LOQ: Limit of quantitation, S/N: Signal-to-noise ratio, RSD: Relative standard deviations, DP: Declustering potential, CE: Collision energy, MRM: Multiple reaction monitoring, RT: Retention time.

Background: Curative plants have reportedly been used to make chewing sticks/toothbrushes intended for the treatment of oral diseases. Objective: The in vitro antibacterial activities of Azadirachta indica, Pongamia pinnata, Psidium guajava, and Mangifera indica were evaluated against Streptococcus mutans, along with the cytotoxicity and antioxidant and synergistic potentials. The effect of M. indica on the expression of crucial virulence genes spaP and gtfB of S. mutans was determined. Materials and Methods: The antibacterial activity was determined using a modified microdilution method. The antioxidant potential was evaluated using diphenyl picrylhydrazyl (DPPH), Griess reagent, and nitroblue tetrazolium calorimetric assays. The synergistic activity was investigated using a modified checkerboard method, while the cytotoxicity was determined according to a cell proliferation 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide salt assay. Reverse transcription was the chosen method for determining the difference in expression of the spaP and gtfB genes after treatment with the plant sample. Results: M. indica and A. indica had the highest antibacterial activity at concentrations of 0.3 mg/ml and 6.25 mg/ml, respectively. A. indica had the best free radical scavenging of DPPH, exhibiting 50% inhibition at 28.72 μg/ml; while M. indica showed better superoxide scavenging potential than the positive control quercetin. Both M. indica and A. indica had adequate activity against the nitric oxide-free radical (12.87 and 18.89 μg/ml, respectively). M. indica selectively reduced the expression of the gtfB gene, indicating a mechanism involving Glucotranferases, specifically targeting bacterial attachment. Conclusion: The overall good antibacterial activity of M. indica correlated with the general good antioxidant capacity, while showing a potentially unique mechanism of bacterial inhibition, targeting virulence gene expression. Abbreviations used: AA: Ascorbic acid; BHI: Brain–heart infusion; CHX: Chlorhexidine; DPPH: Diphenyl picrylhydrazyl; DMSO: Dimethlysulfoxide; NBT: Nitroblue tetrazolium; NO: Nitric oxide; XTT: 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5 -carboxanilide salt.

Background: Addiction to ketamine is becoming a serious public health issues, for which there exists no effective treatment. Rhynchophylline (Rhy) is an alkaloid extracted from certain Uncaria species that is well known for both its potent anti-addictive and neuroprotective properties. Increasing evidence supports the contributions of cAMP response element binding protein (CREB), nuclear receptor-related-1 (Nurr1), and brain-derived neurotrophic factor (BDNF) in modulating neural and behavioral plasticity which was induced by addictive drugs. Objective: To investigate the effects of Rhy on the behavior and the levels of phosphorylated CREB (p-CREB), Nurr1, and BDNF in the hippocampus of ketamine-induced conditioned place preference (CPP) rats. Materials and Methods: CPP paradigm was used to establish the model of ketamine-dependent rats and to evaluate the effect of Rhy on ketamine dependence. The expressions of p-CREB, Nurr1, and BDNF were tested by Western blotting and immunohistochemistry. Results: We observed that Rhy can reverse the behavior preference induced by ketamine CPP training. At the same time, expression of p-CREB, Nurr1, and BDNF, which was significantly increased by ketamine, was restored in the Rhy -treated group. Conclusion: This study indicates that Rhy can reverse the reward effect induced by ketamine in rats and the mechanism can probably be related to regulate the hippocampal protein expression of p-CREB, Nurr1, and BDNF. Abbreviations used: Rhy: Rhynchophylline; CREB: cAMP response element binding protein; Nurr1: Nuclear receptor-related-1; BDNF: Brain-derived neurotrophic factor; CPP: Conditioned place preference; NMDA: N-methyl-D-aspartic acid; METH: Methamphetamine; CNS: Central nervous system; PFA: Paraformaldehyde; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; LTP: long-term potentiation.

Background: Arctigenin has many pharmacological activities with clinical significance and is derived from Arctium lappa L. However, the present extraction method is inefficient and does not have meaningful industrial production. Objective: A new method to directly prepare arctigenin was established by combining enzyme-assisted extraction and central composite design. Arctigenin's further pharmacological activity was also surveyed in vitro. Materials and Methods: β-D-Glucosidase, a food-grade enzyme, was added directly to the fruits of A. lappa L. to hydrolyze the arctiin to arctigenin, and the obtained samples were subsequently subjected to ethanol (30%, v/v) extraction. The pharmacological activity of the extraction and arctigenin was determined by inhibiting acetylcholinesterase (AChE) and scavenging nitrite. Results: The factors investigated include the enzyme concentration (0.5%–2.5%), ultrasound time (10 min⁻³ 0 min), and extraction temperature (30°C–50°C). From the analysis of the results by Design-Expert (V8.0.6), the optimal extraction conditions were obtained: enzyme concentration (1.4%), ultrasound time (25 min), and extraction temperature (45°C). The highest yield of arctigenin, obtained under the optimal conditions was 6.39%, representing an increase of 28.15% compared to the reference extraction without enzyme processing. The IC₅₀values of the extraction and arctigenin, respectively, for inhibiting AChE were 0.572 mg/ml and 0.462 mg/ml, and those for nitrite-scavenging were 34.571 mg/ml and 17.49 mg/ml. Conclusions: The results demonstrate that using an enzyme directly in the production is an effective means for extracting arctigenin from Fructus arctii. The extraction has the activities of inhibiting AChE and scavenging nitrite, probably because there has arctigenin in it. It is implied that the extraction and arctigenin could contribute to human health in clinical applications. Abbreviations used: AChE: Acetylcholinesterase, CCD: Central composite design, TCM: Traditional Chinese medicines, AD: Alzheimer's disease, DTNB: 5,5'-dithiobis-(2-nitrobenzoic acid), DMSO: Dimethyl sulfoxide, ATCI: Acetylthiocholine iodide.

Background: Tripterygium wilfordii (TW) is widely employed to treat rheumatoid arthritis and autoimmune disorders clinically, which, however, accompany with disturbing hepatotoxicity and nephrotoxicity. The previous research showed that Panax notoginseng (PN) compatibly and significantly reduces the TW-induced hepatotoxicity. Objective: To explore the underlying mechanism, the present study was designed to reveal the influence of PN on the intestinal absorption process of TW-derived active components in rat. Materials and Methods: An in situ single-pass intestinal perfusion technique was established and preformed to obtain the perfusate samples of triptolide (TP), tripterine (TE), TW extract, and TW-PN extract. A rapid and sensitive ultra-performance liquid-chromatography tandem mass spectrometry method was subsequently developed and validated to determine the concentrations of TP and TE in the perfusate samples. Then, the absorption parameters, effective permeability, absorption rate constant, and percentage of 10 cm intestinal absorption were calculated strictly. Results: The final data indicated that both TP and TE have no special absorption site in the intestine and are primarily absorbed in a passive manner. Otherwise, the absorption of TP was decreased from compatibility of PN, but the absorption of TE was enhanced. Conclusion: The absorption reduction of TP and absorption elevation of TE from TW initiated by the combination of PN are contributed to attenuate the toxicity and reinforce the therapeutic efficacy of TW. It is practically reasonable of usage of TW compatibility with PN clinically. Abbreviations used: 10 cm% ABS: percentage of 10 cm intestinal absorption, DMARDs: Disease-modifying antirheumatic drugs, GU: Glycyrrhiza uralensis, K_a: Absorption rate constant, NSAIDs: Nonsteroidal anti-inflammatory drugs, P_eff: Effective permeability, PN: Panax notoginseng, QC: Quality control, RA: Rheumatoid arthritis, RG: Rehmannia glutinosa, SPIP: Single-pass intestinal perfusion, TE: Tripterine, TP: Triptolide, TW: Tripterygium wilfordii, UPLC-MS/MS: Ultra-performance liquid-chromatography tandem mass spectrometry.

Background: Mulberry fruits are a superior source of polyphenol, especially anthocyanins that contribute potentially to the beneficial effects which include reducing the risk of cardiovascular diseases and cancers with antioxidant, anti-inflammatory, and chemoprotective properties. Objectives: In this study, purification of the polyphenol-rich extract from mulberry fruit (MPE) was purified and assessed the activities of antioxidant and hemolysis protective in vivo and in vitro. Materials and Methods: Antioxidant activities in vitro was measured by quantifying its 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity, reducing power and Fe²⁺-chelating ability. MPE was purified by high-pressure liquid chromatography (HPLC) and analyzed individual polyphenols using liquid chromatography–mass spectrometry (LC-MS)/MS. Results: The total polyphenol content was 147.69 ± 0.02 mg gallic acid equivalents (GAE)/g dried weight (DW) in the extract and 403.55 ± 0.02 mg GAE/g DW in the purified extract. Further identification by HPLC-ultraviolet-visible and LC-MS/MS analysis indicated in MPE, an anthocyanin compound, cyanidin-3-O-glucoside. With regard to in vitro assays, MPE possessed antioxidant effect, especially in Fe²⁺ chelating ability with an IC₅₀value of 1.016 mg/mL. The protective effects on mouse red blood cell hemolysis and lipid peroxidation ex vivo were dose and time dependent. Conclusion: It indicates that MPE could be a good candidate for future biomedical applications to promote human health with limited side effects. Abbreviations used: MPE: Purification of the polyphenol-rich extract from mulberry fruit, LC-MS: Liquid chromatography–mass spectrometry, HPLC: High-pressure liquid chromatography, DPPH: 2,2-diphenyl-1-picrylhydrazyl scavenging activity, RBC: Red blood cell, GAE: Gallic acid equivalent, FeCl₂: Ferrous chloride, H₂O₂: Hydrogen peroxide, EDTA-2Na: Ethylenediaminetetraacetic acid disodium salt, PBS: Phosphate-buffered saline, TCA: Trichloroacetic acid, TBA: 2-thiobarbituric acid, FeSO₄: Ferrous sulphate, MDA: Malondialdehyde, V_C: Vitamin C, DW: Dried weight.

Introduction: Melanin is synthesized by melanocytes, which are located in the basal layer of the skin. After synthesis, melanin is further deposited on the surface of the skin to form black spots or chloasma. Tyrosinase is a rate-limiting enzyme that plays an important role in melanogenesis. Currently, there are many drugs that inhibit tyrosinase expression to further reduce melanogenesis. Nevertheless, some of these could reverse the pharmacological effect of other drugs, when used simultaneously. Materials and Methods: B16 mouse melanoma cells were treated with the tyrosinase inhibitors licochalcone A and β-arbutin, alone or in combination with capsaicin, an alkaloid found in peppers. Cytotoxicity, melanin content, and tyrosinase activity and expression were determined. Results: Licochalcone A/β-arbutin inhibited tyrosinase expression and further hindered melanin synthesis when applied individually to B16 mouse melanoma cells. However, licochalcone A/β-arbutin combined with 50 μmol/L capsaicin enhanced the expression of tyrosinase in these cells and further increased melanin content. Conclusion: Our data implied that capsaicin could reverse the inhibitory effect of licochalcone A/β-arbutin on tyrosinase expression in B16 mouse melanoma cells.Abbreviations used: B16: B16 mouse melanoma cells; L-DOPA: 3, 4-L-dihydroxyphenylalanine; TYR: Tyrosinase; USP: United States Pharmacopeia; FBS: Fetal bovine serum; EDTA: Ethylenediaminetetraacetic acid; DMSO: Dimethyl sulfoxide; RPMI: Roswell Park Memorial Institute; MTT3: 4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, NaOH: Sodium hydroxide; PBS: Phosphate-buffered saline; RIPA: Radio-immunoprecipitation assay; PMSF: Phenylmethanesulfonyl fluoride or phenylmethylsulfonyl fluoride; SDS: Sodium dodecyl sulfate, sodium salt; PVDF: Polyvinylidene fluoride; ECL: Enhanced chemiluminescence.

Background: Cordyceps militaris fraction (CMF) has been shown to possess in vitro antitumor activity against human chronic myeloid leukemia K562 cells in our previous research. Materials and Methods: The in vitro inhibitory activities of CMF on the growth of KB cells were evaluated by viability assay. The apoptotic and cell cycle influences of CMF were detected by 4′,6-diamidino-2-phenylindole staining and flow cytometry assay. The expression of different apoptosis-associated proteins and cell cycle regulatory proteins was examined by Western blot assay. The nuclear localization of c-Jun was observed by fluorescence staining. Objective: The objective of this study was to investigate the antiproliferative effect of CMF as well as the mechanism underlying the apoptosis and cell cycle arrest it induces in KB cells. Results: CMF suppressed KB cells' proliferation in a dose- and time-dependent manner. Flow cytometric analysis indicated that CMF induced G2/M cell cycle arrest and apoptosis. Western blot analysis revealed that CMF induced caspase-3, caspase-9, and PARP cleavages, and increased the Bax/Bcl-2 ratio. CMF also led to increased expression of p21, decreased expression of cyclin B1, mitotic phosphatase cdc25c, and mitotic kinase cdc2, as well as unchanged expression of p53. In addition, CMF stimulated c-Jun N-terminal kinases (JNK) protein phosphorylations, resulting in upregulated expression of c-Jun and nuclear localization of c-Jun. Pretreatment with JNK inhibitor SP600125 suppressed CMF-induced apoptosis and G2/M arrest. Conclusions: CMF is capable of modulating c-Jun caspase and Bcl-2 family proteins through JNK-dependent apoptosis, which results in G2/M phase arrest in KB cells. CMF could be developed as a promising candidate for the new antitumor agents. Abbreviations used: CMF: Cordyceps militaris fraction; OSCC: Oral squamous cell carcinoma; JNK: c-Jun N-terminal kinase.

Background: Pyrethroids have prominently known for their insecticidal actions worldwide, but recent reports as anticancer and antiviral applications gained a lot of interest to further understand their safety and immunotoxicity. Objective: This encouraged us to carry out our present study to evaluate the interactions of pyrethroids toward adaptive immune cell receptors. Materials and Methods: Type 1 and Type 2 pyrethroids were tested on T (CD4 and CD8) and B (CD28 and CD45) immune cell receptors using Maestro 9.3 (Schrödinger, LLC, Cambridge, USA). In addition, top-ranked tested ligands were too explored for toxicity prediction in rodents using ProTOX tool. Results: Pyrethroids (specifically type 2) such as fenvalerate (−5.534 kcal/mol: CD8), fluvalinate (−4.644 and − 4.431 kcal/mol: CD4 and CD45), and cypermethrin (−3.535 kcal/mol: CD28) have outcome in less energy or more affinity for B-cell and T-cell immune receptors which may later result in the immunosuppressive and hypersensitivity reactions. Conclusion: The current findings have uncovered that there is a further need to assess the Type 2 pyrethroids with wet laboratory experiments to understand the chemical nature of pyrethroid-induced immunotoxicity. Abbreviations used: PDB: Protein Data Bank; AOFA: Amine oxidase (flavin-containing) A; PGH 1: Prostaglandin G/H synthase 1.

Aim: The aim of this study is to establish a method for determination of eupatilin in Folium artemisiae Argyi and observe the inhibitory effect of Folium artemisiae Argyi extract on human hepatoma SMMC-7721 cells. Methods: High-performance liquid chromatograph system with 2910 pump, 2930 UV detector, and N2000 workstation was used for determination of eupatilin in Folium artemisiae Argyi. Human hepatoma SMMC-7721 cells were cultured and cell proliferation was measured using the MTT assay. The expression protein levels of p53, Topo II, and bcl-2 were detected using Western blotting. Results: Eupatilin exhibited a linearity range of 0.5–3.0 μg/mL and a recovery of 100.72%, relevant standard derivation = 2.28%. Folium artemisiae Argyi extract had marked cytostatic and cytotoxic effects on SMMC-7721 cells, inhibited the SMMC-7721 colony formation in a dose-dependent manner. Folium artemisiae Argyi extract already possessed delayed effect after treating SMMC-7721 cells for 8 h, which became obvious at 12 h from treatment. After drug withdrawal, cells still tended to apoptosis. Folium artemisiae Argyi extract could inhibit p53, Topo II, and bcl-2 expressions in tumor cells. The present method for determination of eupatilin is simple, fast, accurate, sensitive, and reproducible. Conclusion: Hepatoma SMMC-7721 cells are quite sensitive to Folium artemisiae Argyi extract, which may be associated with its suppression of p53, Topo II, and bcl-2 expressions. Abbreviations used: HPLC: High-performance liquid chromatograph; OD: Optical density; RSD: Relevant standard derivation; IC₅₀: Inhibitory 50% concentration.

Background: Rhubarb is a traditional Chinese medicine for treating traumatic brain injury (TBI). Purpose: The purpose of this study is to elucidate the potential mechanism of rhubarb by suppressing extracellular signal-regulated kinase (ERK) to ameliorate brain edema. Materials and Methods: Sprague-Dawley rats were separated into four groups at random. One group received 3 g/kg rhubarb, and another group received 12 g/kg rhubarb, and the vehicle group and sham group were administered the same dose of saline solution. The blood–brain barrier disruption and edema were detected by Evans blue extravasation and water content, respectively. ERK, Matrix metalloproteinase 9 (MMP-9), and zonula occluden-1 (ZO-1) in the damaged tissue were measured by western blot analysis and quantitative real-time polymerase chain reaction. Results: Rhubarb attenuated the brain edema after TBI, especially at the dose of 12 g/kg. Rhubarb significantly suppressed ERK, down-regulated MMP-9, and up-regulated ZO-1. Rhubarb might be a prospective therapeutic regimen to decrease edema in TBI. Conclusions: Rhubarb alleviates the edema by restraining the ERK signaling pathway. Our results contribute to the validation of the traditional use of rhubarb in the treatment of TBI and its mechanism. Abbreviations used: TBI: Traumatic brain injury, ERK: Extracellular signal-regulated kinase pathway, MMP-9: Matrix metalloproteinase 9, ZO-1: Zonula occluden-1, BBB: Blood-brain barrier, PCR: Polymerase chain reaction, TCM: Traditional Chinese medicine, MAPKs: Mitogen-activated protein kinases, CCI: Controlled cortical impact, DL: Rhubarb 3 g/kg in distilled water, DH: Rhubarb 12 g/kg in distilled water, EB: Evans blue, IOD: Integral optical density, MEK: Mitogen extracellular kinase, MMPs: Matrix metalloproteinases, NADPH: Nicotinamide adenine dinucleotide phosphate: ROS, reactive oxygen species.