Acute and subchronic toxicity as well as evaluation of safety pharmacology of modified pulsatilla granules

The present study investigated acute and subchronic toxicity and safety pharmacology of modified pulsatilla granules（MPG）to provide a basis for a comprehensive understanding of MPG toxicity.The results of acute toxicity testing showed that the median lethal dose of MPG was more than 5 000 mg kg^-1,suggesting that MPG was considered as practically non-toxic.The subchronic toxicity study for 30 days was conducted by daily oral administration at doses of 375,750 and 1 500 mg kg^-1 in Sprague-Dawley rats.The results of subchronic toxicity study showed that the body weight and relative organ weight were not significantly changed by administration of MPG.The clinical chemistry study showed that MPG could induce kidney and liver damages.In histopathological,mild lesions in liver and kidney were also observed,suggesting that the liver and kidney might be potential target organs of MPG.In the safety pharmacology study,MPG did not exhibited any side effects to rats in cardiovascular system,respiratory system and central nervous system.These results suggested that MPG could be considered safe for veterinary use.

Modified Liangfu granule exhibits anti-cancer effects in gastric cancer by regulating apoptosis-related proteins and genes

Objective:To investigate the anti-cancer effects and mechanism of modified Liangfu granule(MLFG)in BGC-823 gastric cancer cells.Methods:The drug serum was extracted from abdominal aorta of Wistar rats treated by cyclophosphamide,dioscin,MLFG(high-,medium-,and low-dose),respectively.MTS assay was performed to detect the effect of different concentrations of MLFG and dioscin on cell growth.The effect of MLFG on cellular apoptosis was detected by flow cytometry.Western blot was used to examine Bcl-2 and Akt in BGC-823 cells treated with MLFG.Quantitative real-time polymerase chain reaction assay was performed to determine the expression level of caspase-3,E2F1 and E2F3 genes in cells treated by MLFG-and dioscin-containing serum.Results:MLFG-and dioscin-containing serum inhibited the cell proliferation of BGC-823 cells.MLFG induced gastric carcinoma cell apoptosis and inhibited the expression of Bcl-2 and Akt in a dosedependent manner.MLFG also significantly induced the gene expression of caspase-3 and downregulated E2F1 and E2F3 gene expression.Conclusion:The effect of MLFG and dioscin on inhibiting cell proliferation and induction of apoptosis of gastric cancer cells might be related to the regulation of Bcl-2 and Akt proteins and the expression of caspase-3,E2F1 and E2F3 genes.

Modified Sijunzi Granules Exhibit Hemostatic Effect by Activating Akt and Erk Signal Pathways via Regulating 5-HT and Its Receptors Levels

Objective:To investigate the hemostatic effect of modified Sijunzi Granules(MSG)in primary immune thrombocytopenia(ITP)zebrafish model and explore the potential mechanism.Methods:AB strain wild type zebrafish were treated with simvastatin(6μmol/L)for 24 h to establish the hemorrhage model(model control group).The zebrafish were treated with MSG at different doses(55.6,167,and 500μg/mL),respectively.The hemostatic effect was assessed by examining the intestinal bleeding and hemostatic rate.5-hydroxytryptamine(5-HT)content was determined using enzyme-linked immunosorbent assay(ELISA)assay.The expressions of5-HT2aR,5-HT2bR,and SERT genes were detected by quantitative real-time polymerase chain reaction(PCR).The protein expressions of protein kinase B(Akt),p-Akt,extracellular regulated protein kinases(Erk),and p-Erk were examined using Western blot analysis.Results:The intestinal bleeding rate was 37%,40%,and 80%in the55.6,167,and 500μg/mL dose of MSG,respectively,in which 55.6 and 167μg/mL MSG dose groups were associated with significantly decreased intestinal bleeding rate when compared with the model control group(70%,P<0.05).Significantly higher hemostatic rates were also observed in the 55.6μg/mL(54%)and 167μg/mL(52%)MSG dose groups(P<0.05).MSG increased the 5-HT content and mRNA expression levels of 5-HT2aR,5-HT2bR,and SERT(P<0.05).In addition,caspase3/7 activity was inhibited(P<0.05).Significant increase in p-Akt and p-Erk was also detected after treatment with MSG(P<0.05).Conclusions:MSG could reduce the incidence and severity of intestinal bleeding in zebrafish by activating MAPK/Erk and PI3K/Akt signal pathways through regulating the levels of 5-HT and its receptors,which may provide evidence for the treatment of ITP.

Efficacy of Modified Wuzhuyu Decoction Granule (加减吴茱萸汤颗粒剂) for Migraine Patients with Cold and Stasis Obstructing Meridian Syndrome： A Randomized, Double-Blind, Placebo-Controlled Trial

Objective： To study the efficacy of modified Wuzhuyu Decoction Granule （加减吴茱萸汤颗粒剂, MWDG） in the treatment of migraine patients with cold and stasis obstructing meridian syndrome. Methods： This study was a randomized, double-blind, placebo-controlled trial. A total of 78 migraine patients with cold and stasis obstructing meridian syndrome were recruited and randomly assigned by a ratio of 2：1 into a treatment group （51 cases） and a placebo group （27 cases）. Patients in the treatment group were treated with MWDG while placebo granules were applied in the control group. The treatment course lasted for 12 weeks with a follow-up of 4 weeks. The primary outcome measures included frequency and days of migraine attacks and the secondary outcome measures were analgesics consumption and visual analogue scale （VAS） scores. All outcome assessments were conducted respectively at baseline, the 4th, 8th and 12th week, and the end of follow-up. Results： In the treatment group, significant decrease in frequency of migraine attacks were observed since the 4th week and that of analgesics consumption since the 8th week （both P〈0.05）. While, in the placebo group, significant decrease in frequency of migraine attacks were observed since the 8th week and that of analgesics consumption since the 12th week （both P〈0.05）. No significant decrease in days of migraine attacks and VAS scores of migraine pain were observed in both groups. Between the two groups, there were significant differences in VAS scores and intensity of pain appeared in the 8th week （P〈0.05）. However, no significant differences were found in days and frequency of migraine attacks and analgesics consumption （P〉0.05）. Conclusions： MWDG was probably effective in the treatment of migraine especially for alleviating pain intensity. Furthermore, MWDG could reduce the frequency of migraine attacks and analgesics consumption sooner than the placebo.

Effect of Modified Wuzi Yanzong Granule(加味五子衍宗颗粒)on Patients with Mild Cognitive Impairment from Oxidative Damage Aspect

Objective： To observe the effects of modified Wuzi Yanzong Granule （加味五子衍宗颗粒,WYG） on memory function and the activity of serum superoxide dismutase （SOD）, malondialdehyde （MDA） levels, leukocyte mitochondrial DNA （mtDNA） deletion rate and β-amyloid protein1.28 （A β 1-28 ） in patients with mild cognitive impairment （MCl）. Methods： Thirty-six patients with MCl were selected based on the internationally recognized Petersen＇s criteria, and equally and randomly assigned to two groups. The treated group was treated with WYG and the control group was treated with placebo for 3 months. In addition, 20 healthy subjects were included in the study as the normal control group. Changes of memory function, SOD activity, MDA content, leukocyte mtDNA deletion rate and A β 1-28 content were observed before and after treatment. Results： Compared with the normal control group, the memory quotient and SOD activity in patients with MCl decreased significantly （P 〈 0.01）, while MDA, A β1-28 levels and the leukocyte mtDNA deletion rate increased significantly （P 〈 0.01）. After treatment, levels of memory quotient and serum SOD activity increased while the serum MDA level, leukocyte mtDNA deletion rate and A β 1-28 level decreased in the treated group compared with those before treatment （P〈0.01, P〈0.05）. Meanwhile, leukocyte mtDNA deletion rate and A β 1-28 content in the treated group were all lower than those in the control group （P〈0.05）. Conclusion： WYG could improve memory function in patients with MCl and the therapeutic mechanism is possibly related to the increased activity of anti-oxidase, the improved free radical metabolism and the alleviation of leukocyte mtDNA oxidation damage. WYG shows clinical significance in delaying the progression of MCl.