Effect of Yiqi Bushen Granule (益气补肾颗粒) on the Peripheral Natural Killer Cell and γ δ T-cell in the Patients with Minimal Residual Leukemia

Objective： To analyze the changes in peripheral natural killer T-cells （NKT） and gammadelta T-cells （γδT-cell） in patients with minimal residual leukemia （MRL） before and after being treated with Yiqi Bushen Granule （益气补肾颗粒, YBG） in order to determine their significance in prognosis of the disease. Methods： Before and after treatment, the changes in 36 patients （16 males and 20 females） receiving long-term （more than 3 months） YBG therapy were analyzed using multi-parameter flow cytometry, with 34 healthy persons （19 males and 15 females） acting as controls. Results： The absolute value and percentage of NKT cells and γδT-cells were all significantly raised after treatment, for NKT cells, 0.52%±0.39% to 0.83%±0.66% and 7.25±7.77 cell/μL to 12.86±11.99 cell/μL, for γδT-cells, 6.08%±3.03% to 7.24%±2.78% and 83.97±48.09 cell/μL to 110.53±54.12 cell/μL, respectively （P0.05 or P0.01）. Conclusion： YBG could regulate the immune function and elevate the amount of NKT cells and γδT-cells, thus to kill or suppress the residual leukemic cell in the body, which might be one of the mechanisms of YBG in prolonging the disease-free survival in MRL patients.

Network pharmacology-based prediction and verification of the mechanism for Bushen Chengyun granule on low endometrial receptivity

Objective:Bushen Chengyun granule(BCG)is an empirical treatment for female infertility(FI)caused by low endometrial receptivity(LER)involving a poorly understood mechanism.In this study,network pharmacology was used to explore the potential therapeutic mechanism of BCG on FI caused by LER.Methods:The corresponding herb targets were obtained by conducting a search in the Traditional Chinese Medicine Systems Pharmacology Database and PubMed-reported literature.Disease targets were obtained from the following databases:Comparative Toxicogenomics Database,Human Phenotype Ontology,and Therapeutic Target Database.Treatments for LER using BCG have used target matching(BCG e LER target).Then,the predicted targets were uploaded to the Search Tool for the Retrieval of Interacting Genes/Proteins database for gene ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses.Furthermore,triptorelin acetate for injection t menotrophin t chorionic gonadotropin for injection were used to establish a mouse model of blastocyst implantation disorder and to evaluate the in vivo effect of BCG on blastocyst implantation.Results:Overall,156 bioactive chemical components and 1092 targets of BCG were identified.The results indicated that 482 biological processes(FDR<0.01)and 15 pathways(FDR<0.01)related to BCG participated in the complex treatment effects and were associated with the endocrine system,inflammatory responses,metabolism,apoptosis,ovulatory performance,and angiogenesis.Moreover,16 hub nodes of BCG including estrogen receptor(ESR1),estrogen receptor beta(ESR2),progesterone receptor,et al,were recognized as potential treatment targets and might help clarify the underlying therapeutic mechanisms of BCG for female infertility.BCG significantly increased the protein expressions of estrogen receptors and progesterone receptors.Conclusions:These findings reveal the potential therapeutic mechanism of BCG for female infertility involves low endometrial receptivity,which should be evaluated further.

VEGFR2 mediated RAS/MAPK signaling pathway to explore the mechanism of Bushen Antai Granule in the treatment of recurrent abortion

Objective:To investigate the effect of Bushen Antai Granule on the mRNA and protein expression of Ras protein/mitogen activated protein kinase mediated by vascular endothelial growth factor receptor 2 with small interference RNA interference.Methods:Method to construct the placenta microvascular endothelial cells,and the preparation of kidney fetus granule drug-containing serum,select the best drug-containing serum concentration,it can be divided into normal group,the serum siRNA-NC normal serum group,drug serum,siRNA normal serum group,siRNA drug serum group,using real-time fluorescent quantitative PCR,Western blotting,immunofluorescence test respectively the RAS/MAPK mRNA and protein expression.Results:Results there was no significant difference in Ras and MAPK mRNA and protein expression between the normal group and the negative control group(P>0.05).The mRNA and protein expressions of Ras and MAPK in the drug serum group were significantly higher than those in the normal serum group(P<0.01).Ras and MAPK mRNA and protein expression were significantly decreased in siRNA1 normal serum group compared with normal serum group(P<0.01).Ras,MAPK mRNA and protein expression in siRNA1 drug serum group were significantly different from that in siRNA1 normal serum group(P<0.01).Conclusion:Conclusion The therapeutic effect of Bushen Antai Granule on recurrent abortion may be realized by upregulation of RAS/MAPK mRNA and protein expression.

Mechanism of Bushen Tongluo granule on osteoarthritis based on network pharmacology and molecular docking technology

Objective:In this study,we used network pharmacology and molecular docking technology to analyze the mechanism of Bushen Tongluo granule in the treatment of osteoarthritis.Methods:The main active components and corresponding targets of Bushen Tongluo granule were screened from thetraditional Chinese medicine systems pharmacology database.The targets related to osteoarthritis were collected from the Online Mendelian Inheritance in Man,Therapeutic Target database,GeneCards,Pharmacogenomics Knowledgebases and Drugbank databases.Cytoscape3.9.0 software was used to construct the action network diagram of“Bushen Tongluo Granule-Active Component-Target”.Builded a protein-protein interaction network from the STRING database.The Bioconductor platform and R language 4.0.3 were used for Gene Ontologyfunction andKyoto Encyclopedia of Genes and Genomespathway enrichment analysis.Then,selected the pathway most associated with osteoarthritis for specific analysis.Finally,the core genes were screened and verified by molecular docking using AutoDockTools software.Results:71 principal components of Bushen Tongluo granule and 183 potential therapeu-tic targets for osteoarthritis were obtained.Twenty-eight key targets of Bushen Tongluo granulein the treatment of osteoarthritis are enriched in 158 pathways.Among them,the tumor necrosis factor signaling pathway,interleukin-17 signaling pathway,Toll-like receptor signaling pathway,T helper cell 17 cell differentiation and hypoxia-inducible factor-1signaling pathway involved in key targets are closely related to osteoarthritis.The relevant vital targets were involved in the regulation of DNA transcription factor activity,the response to chemical stress,the response to reactive oxygen species,the response to oxidative stress,the proliferation of muscle cells,the proliferation of epithelial cells and the biological processes such as responses to lipopolysaccharides,responses to molecules of bacterial origin.Molecular docking showed that protein kinase B 1-β-sitosterol,the tumor necrosis factor-naringenin,interleukin-6-luteolin,mitogen-activated protein kinase-quercetin,vascular endothelial growth factor A-quercetin and prostaglandin-endoperoxide synthase 2-luteolin have strong docking activities.Conclusions:Bushen Tongluo granule can reduce the inflammatory response and inhibit articular cartilage angiogenesis in the treatment of osteoarthritis,which may be achieved by regulating the inflammatory signaling pathway and hypoxia-inducible factor-1 signaling pathway.

Bushen Qiangji Granule (补肾强脊颗粒) Medicated Serum Inhibits Osteogenic Differentiation of Fibroblasts in AnkylosingSpondylitis by Inhibiting the BMP/Smads Signal Pathway in vitro

Objective: To explore the mechanism of Bushen Qiangji Granule(补肾强脊颗粒, BSQJ) in restraining the osteogenic differentiation of ankylosing spondylitis(AS) fibroblasts. Methods: Hip joint capsules were obtained from AS patients(n=10) receiving total hip replacement and healthy hip joint capsules from patients with hip fracture(n=10) receiving surgery as a control. Finite fibroblast lines were established from these tissue samples to observe the effect of BSQJ on suppressing osteogenic differentiation of fibroblasts. The expression of osteogenic marker gene corebinding factor a1(Cbfa1) and Smad family proteins were examined by Western blot and real-time quantitative polymerase chain reaction(q PCR). Results: The m RNA expression level of Cbfa1 was significantly higher in AS fibroblasts than that in normal fibroblasts and the expression of p Smad1, p Smad5, Smad4 and Cbfa1 in AS fibroblasts was also higher, demonstrating the activation of the BMP/Smads signal pathway in AS fibroblasts. BSQJ-medicated serum not only restrained the m RNA and protein expression levels of Cbfa1 and inhibited protein expression level of Smad4 but also decreased the expression quantities of p Smad1 and p Smad5. Conclusions: BSQJ can inhibit osteogenic differentiation of AS fibroblasts in vitro by suppressing the activation of the BMP/Smads signal pathway. This may be the important molecular mechanism of BSQJ in regulating AS ossification.

Effectiveness of Bushen Huoxue Granule (补肾活血颗粒) on 5-Serotonin and Norepinephrine in the Brain of Parkinson's Disease Patients with Depressive State

Objective：To observe the clinical curative effect of Chinese medicine Bushen Huoxue Granule（补肾活血颗粒,BHG） on Parkinson＇s disease（PD） patients with depressive state.Methods：Sixty-two PD patients with depressive state were randomly assigned to two groups by using a random number table,31 in each group.Madopar was given to all as the conventional treatment.The fluoxetine hydrochloride dispersible tablet was given to the patients in the control group and BHG was given to those in the treatment group.The therapeutic course for all was 12 weeks.Before and after treatment,Hamilton depression rating scale（HAMD） was applied to judge the curative effect,and the changes of cerebral neurotransmitters levels in the brain of patients were detected by encephalofluctuograph technique.Results：The scores of HAMD in the two groups were decreased markedly after 12-week treatment.It was lower in the treatment group than that in the control group with significant difference（P〈0.01）.The contents of norepinephrine（NE） and 5-serotonin（5-HT） in the PD patients were obviously lower than normal value.There was no significant difference between the two groups before treatment（P〉0.05）.The contents of NE and 5-HT were all increased in the two groups after treatment（P〈0.05）,with significant differences between the two groups（P〈0.01）.Conclusion：BHG could increase the contents of NE and 5-HT in PD patients＇ brain to improve the depressive state of PD patients.

Study on Mechanism of BushenYugong Granule(补肾育宫冲剂) inTreating Uterine Dysgenesia

Objective: To explore the therapeutic mechanism of Bushen Yugong Granule (BSYGG, a Chinese medicinal preparation) in treating infertility due to uterine dysgenesia. Methods: (1) Clinical study: Serum level of sex hormone and endometrial contents of estradial receptor (ER) and progesterone receptor (PR) in follicle period of 60 patients (the tested group) were measured and compared before and after treatment, and also compared with those of 70 normal women (the control group). (2) Experimental study: Rats were divided into the control group and 3 tested groups fed with different doses of BSYGG. Pathologic examination of pituitary, ovary and uterus of rats after BSYGG treatment were performed,and serum levels of ER, PR measu red. Also, an estrogen activity experiment was conducted in some young oophorectomized rats. Results: (1) Clinical study showed the difference of serum sex hormone levels between the tested and control group was insignificant. The percentage of patients in the tested groups with endometrial ER content over was lower than that in the control group (46.9% vs 72.4%) significantly (P<0.05), after treatment, it rose to 65.4%,a nd the change was significant, P<0.05. (2) Experimental study showed that the number of follicle, weight of uterus and serum E2 level increased significantly after BSYGG treatment. The endometrial content of ER in the control group was 29.25±17.03%, while in the tested groups, it was 67.75±24.56% to 83.00±9.87% after treatment, the difference between the two groups was significant (P<0.005). Conclusion: BSYGG has an estrogen-like activity, it could increase the endometrial content of ER, enhance the effect of estrogen on uterus and promote the development of uterus, so as to elevate fertilization rate.

Chinese Medicine Sequential Therapy Improves Pregnancy Outcomes after Surgery for Endometriosis-Associated Infertility: A Multicenter Randomized Double-blind Placebo Parallel Controlled Clinical Trial

Objective:To evaluate the efficacy and safety of Chinese medicine(CM)improving pregnancy outcomes after surgery for endometriosis-associated infertility.Methods:A multicenter,randomized,doubleblind placebo parallel controlled clinical trial was designed.A total of 202 patients who had laparoscopy for endometriosis-associated infertility with qi stagnation and blood stasis syndrome were included and randomly divided into the CM treatment group and placebo control group at a ratio of 1:1 using a central block randomization from May 2014 to September 2017,101 patients in each group.The two groups received continuous intervention at 1–5 days after surgery,for 6 menstrual cycles.Before ovulation,the CM group was treated Huoxue Xiaoyi Granule(活血消异颗粒);after ovulation,Bushen Zhuyun Granule(补肾助孕颗粒)was involved.The control group was treated with placebo.Transvaginal ultrasonography was performed every menstrual cycle during the treatment,and female hormone levels in the follicular and luteal phases were measured during the 1 st,3 rd and 6 th menstrual cycles.The analysis was continued until pregnancy.The primary outcomes were clinical pregnancy rate and pregnancy outcome,and the secondary outcomes were follicular development and endometrial receptivity.Safety evaluations were performed before and after treatment.Results:(1)Clinical pregnancy and live birth rates:the clinical pregnancy and live birth rates of the CM group were significantly higher than those of the placebo group[44.6%(45/101)vs.29.7%(30/101),34.7%(35/101)vs.20.8%(21/101),both P<0.05].(2)Follicle development:the incidence of dominant follicles,rate of cumulative cycle ovulation,and rate of cumulative cycle mature follicle ovulation were significantly higher in the CM group than those in the placebo group[93.8%(350/373)vs.89.5%(341/381),80.4%(275/342)vs.69.1%(253/366),65.8%(181/275)vs 56.1%(142/253),P<0.05 or P<0.01]).The incidence of cumulative cycle luteinized unruptured follicle syndrome was significantly lower in the CM group than in the placebo group[11.7%(40/342)vs.17.8%(65/366),P<0.05).(3)Endometrial receptivity:after treatment,both endometrial types and endometrial blood flow types in the CM group were mainly types A and B,while those in the placebo group were mainly types B and C,with a significant difference between the two groups(both P<0.05).(4)Adverse events:the incidence of adverse events between the two groups was not significantly different(P>0.05).Conclusion:Strategies for activating blood circulation-regulating Gan(Liver)-tonifying Shen(Kidney)sequential therapy can effectively improve the clinical pregnancy rate and live birth rate of endometriosis-associated infertility with qi stagnation and blood stasis after laparoscopy,improve follicular development,promote ovulation,improve endometrial receptivity,while being a safe treatment option.(Trial registration No.NCT02676713).