Mechanism of Huatan Sanjie Fang in improving goiter in Graves'disease mice based on the Hippo signaling pathway

Objective:To explore the mechanism of Huatan Sanjie Fang(HTSJ)in regulating goiter in Graves'disease(GD)mice by detecting key factors of the Hippo signaling pathway.Methods:A mouse model of GD was established by injecting Ad-TSHR289 adenovirus into the bilateral quadriceps femoris of female mice.Successful mouse models were then randomly divided into a model group,methimazole(MMI)group,and HTSJ group,and fed with deionized water,MMI(4.5 mg/kg per day),and HTSJ(35.10 g/kg per day),respectively,for 10 weeks.Histopathological changes of the thyroid gland were subsequently observed by hematoxylin-eosin staining.Radioimmunoassay was used to detect serum total thyroxine(T4)and thyrotrophin-receptor antibody(TRAb)levels.The relative expression of mRNA of Mst1,YAP,and TAZ were detected by quantitative real-time polymerase chain reaction,while the protein expression of Mst1,YAP,TAZ,pMst1,and pYAP were detected by western blot.Results:After 10 weeks of drug intervention,goiter and other pathological changes in the HTSJ group significantly improved compared with the model group,and the levels of serum T4 and TRAb significantly decreased(P=.002,P<.001,respectively).Decreased mRNA expression of Mst1,YAP,and TAZ,the key factors of the Hippo signaling transduction pathway,was also observed(P=.002,P=.022,P<.001,respectively).In contrast,protein expression of Mst1(P=.046),pMst1(P=.026),and p YAP(P=.004)increased,while protein expression of YAP and TAZ decreased(P=.041,P<.001,respectively).Conclusion:HTSJ can effectively improve goiter in GD mice through the Hippo signaling pathway.

Xiaotan Sanjie decoction attenuates tumor angiogenesis by manipulating Notch-1-regulated proliferation of gastric cancer stem-like cells

AIM: To determine the underlying mechanisms of action and influence of Xiaotan Sanjie (XTSJ) decoction on gastric cancer stem-like cells (GCSCs).

Xiaotan Sanjie decoction inhibits interleukin-8-induced metastatic potency in gastric cancer

AIM:To investigate the interaction between Xiaotan Sanjie(XTSJ) decoction and interleukin-8(IL-8) and its effect on adhesion,migration and invasion of SGC-7901 gastric cancer cells.METHODS:SGC-7901 gastric cancer cells were exposed to serum containing XTSJ decoction and/orIL-8(1 ng/m L).SGC-7901 cell adhesion to fibronectin,an extracellular matrix component,was detected using the Cell Counting Kit-8.Migration and invasion abilities of SGC-7901 cells were detected by scratch wound and Transwell chamber assays.Then,protein(immunofluorescence and Western blot) and m RNA levels(quantitative polymerase chain reaction) of cluster of differentiation 44(CD44),a cell adhesion molecule,were measured in 72-h-cultured SGC-7901 cells.RESULTS:Cell adhesion was promoted by IL-8(P = 0.001),but was inhibited by XTSJ decoction(P = 0.0001).Similarly,IL-8 promoted SGC-7901 cell invasion(P = 0.003),and XTSJ decoction inhibited cell invasion(P = 0.001).IL-8 induced SGC-7901 cell migration,but this was inhibited by XTSJ decoction.IL-8 up-regulated CD44 protein(P = 0.028) and m RNA expression(P = 0.002),whereas XTSJ decoction inhibited CD44 protein expression(P = 0.0001),but not m RNA expression(P = 0.275).An interaction between XTSJ decoction and IL-8 was confirmed in the invasion(P = 0.001) and CD44 m RNA expression of SGC-7901 cells(P = 0.010),but not in cell adhesion(P = 0.051).CONCLUSION:XTSJ decoction may inhibit adhesion,migration and invasion of gastric cancer cells,which is partly associated with down-regulation of IL-8.

Yangzheng Sanjie decoction regulates proliferation and apoptosis of gastric cancer cells by enhancing let-7a expression

AIM To explore the let-7a-mediated anti-cancer effect of Yangzheng Sanjie decoction(YZSJD) in gastric cancer(GC) cells.METHODS YZSJD-containing serum(YCS) was prepared using traditional Chinese medicine serum pharmacology methods. After YCS treatment, cell proliferation and apoptosis were assessed by cell counting kit-8 assay and flow cytometry, respectively, and mi RNA expression profiles were determined using q PCR arrays. Let-7a expression was examined by in situ hybridization in GC tissues and by q PCR in GC cells. c-Myc protein expression was detected by immunohistochemistry in GC tissues, and by Western blot in cell lines.RESULTS YZSJD significantly inhibited proliferation and induced apoptosis in AGS and HS-746 T GC cells. After treatment with YCS, the mi RNA expression profiles were altered and the reduced let-7a levels in both cell lines were up-regulated, accompanied by a decrease in c-Myc expression. Moreover, decreased let-7a expression and increased c-Myc expression were observed during the progression of gastric mucosa cancerization.CONCLUSION YZSJD inhibits proliferation and induces apoptosis of GC cells by restoring the aberrant expression of let-7a and c-Myc.

Rupi Sanjie capsule plus surgical operation for the treatment of hyperplasia nodule of mammary gland:A meta-analysis and trial sequential analysis

Objective:To evaluate the efficacy and safety of Rupi Sanjie(RPSJ)capsule plus the conventional surgical operation for treatment of hyperplasia nodule of mammary gland.Methods:A meta-analysis was conducted on randomized controlled trials(RCT)related to RPSJ capsule plus surgical operation in the treatment of hyperplasia nodule of mammary gland.The methodological quality of eligible RCTs was assessed according to the criteria from the Cochrane Handbook for Systematic Reviews of Interventions.RevMan5.3 and Stata14.0 were used for data analyses.Trial sequential analysis was performed to estimate the sample size of systematic review base on TSA software v0.9.Results:Twenty-two RCTs were totally included in this study,involving 2135 patients.The result showed the clinical recurrence rate of RPSJ capsule plus surgical treatment group was significantly lower than surgical treatment alone group(RR:0.25,95%CI[0.17,0.37],P<0.01).The clinical cure rate and total effective rate were higher in RPSJ capsule plus surgical treatment group than the conventional surgical treatment group(RR:1.63,95%CI[1.46,1.82],P<0.01);(RR:1.29,95%CI[1.22,1.37],P<0.01).Application of RPSJ capsule decreased the occurrence of adverse events including nausea,vomiting,irregular menstruation,constipation,dizziness,and headache etc.(RR:0.90,95%CI[0.54,1.49],P=0.68).The results of trial sequential analysis demonstrated that the current available data did not reach the expected value.Conclusion:RPSJ capsule plus the conventional surgical treatment was more effective in reducing the clinical recurrence rate,and improving the total clinical effective rate and clinical cure rate,with a decrease in the occurrence of adverse events.

Protective effects of Yishen Sanjie Huayu compound on the renal artery disease in rats with IgA nephropathy through ERK/NF-κB pathway

Objective:To observe the effect of Yishen Sanjie Huayu compound prescription on ERK/NF-κB signaling pathway in IgA nephropathy(IgAN)rats,and explore its effect on preventing and treating IgA nephropathy intrarenal arteriole disease.Methods:Fifty-five male SD rats were randomly divided into blank group,model group,ShenfukangⅡcapsule group and Losartan potassium tablet group.Bovine serum albumin(BSA)was used for intragastric administration and carbon tetrachloride(CCl4).IgAN rat model was established by subcutaneous injection and lipopolysaccharide(LPS)tail vein injection.ShenfukangⅡcapsule group and Losartan potassium tablet group were given each drug suspension 2ml/head/d one week after modeling Gavage was started.The blank group and the model group were given an equal volume of normal saline.The 24h urine protein(UTP)of the rats was measured at 4,8,and 12 weeks after the administration,and the blood creatinine(SCr)was measured after 12 weeks.,urea nitrogen(BUN),aldosterone(ADS),angiotensinⅡ(AngⅡ),immunohistochemical Envi-sion System two-step method to detect vascular endothelial growth factor(VEGF)and human matrix in the whole rat kidney and small artery area The expression of metalloproteinase-9(MMP-9),proliferating cell nuclear antigen(PCNA),extracellular regulatory protein kinase(ERK)1/2,nuclear transcription factor-κB(NF-κB),and the small arteries of rat kidney tissue The intima,media,vessel wall/vascular outer diameter value.Results:Compared with the model group,the expressions of VEGF,MMP-9,PCNA,ERK1/2 and NF-κB in kidney tissues of the ShenfukangⅡcapsule group and the Losartan potassium tablet group decreased(P<0.05),24hUTP and SCr,BUN level decreased(P<0.05),kidney tissue damage was alleviated;intima and vessel wall/vascular outer diameter values were significantly reduced(P<0.01),there was no significant difference in ADS between the groups.The AngⅡof the Tanpotassium tablets group was lower than that of the model group(P<0.05).Conclusion:Yishen Sanjie Huayu compound can inhibit the ERK/NF-κB signaling pathway in rats with IgA nephropathy,reduce the levels of VEGF,MMP-9,PCNA,ERK1/2,NF-κB,and inhibit intrarenal arteriole vascular endothelial cells Proliferate and reduce kidney damage.

Effective ingredients, potential targets and mechanism in cancers treatment of Bushen Jiedu Sanjie recipe

Objective: To analyze the active compounds, potential drug targets and corresponding therapy cancer of Bushen Jiedu Sanjie Recipe (BSJDSJR) based on network pharmacology and bioinformatics technology. Methods: The network databases including Cancer HSP, TCMSP, TCMID, TCM-PTD, TCM Database@Taiwan and DrugBank were applied to screen the active compounds, potential drug targets and corresponding cancers of BSJDSJR. Cytoscape3.3 software was used to construct the network between active compounds of Chinese medicine and the corresponding targets. Then, the enrichment of biological processes and KEGG pathways of BSJDSJR were analyzed using DAVID database. Results: According to Oral bioavailability (OB)≥30% and drug like index (DL)≥0.18, 129 active compounds in BSJDSJR were screened out and they targeted to 301 proteins. These targets were closely associated with the occurrence of various cancers, such as bladder cancer, pancreatic cancer, non-small cell lung cancer and colorectal cancer. Further investigation showed that, there were lots of active compounds in BSJDSJR are closely connected with the COX-2/β-catenin signaling pathway, STAT3 signaling pathway and MAPK/ERK signaling pathway. Conclusions: Based on the network pharmacology, the study disclosed the active chemical compounds, potential targets and possible action cancers of BSJDSJR.

＂Impression-Liu Sanjie＂ -A Research on the Participation and Perception of Residents in Scenic Area and Community

Tight space and interactive resources grant tourism community residents a vital body status in community tourism development and construction process. Residents＇ all-sided and effective participation in Tourism development process, with no doubt, is an open democratization process and an effective way to ensure sustainable development of tourism as it can raise residents＇ income and eliminate bad influence of tourism. ＂Impression-Liu Sanjie＂ is a scenic spot where ＂landscape live-action＂ is performed. Local residents （farmers） serve as the main part. Through survey and analysis of participation and perception of community residents, this paper aims at finding some useful consult and inspirations for the development, management and policy formulation of other rural scenic region alike.

Study on the Mechanism of Ruanjian Sanjie Capsules(软坚散结胶囊)Inducing Renal Cancer Cell Apoptosis by Blocking NOTCH Signal Pathway

Objective:To investigate the molecular mechanism of Ruanjian Sanjie Capsules(软坚散结胶囊)through regulating NOTCH signaling pathway to promote human clear cell renal cell carcinoma 7860 cell apoptosis.Methods:A laser confocal microscope was used to observe the nuclear displacement of apoptosis-related proteins,and the expression of Notch1,NICD,Bcl-2,and Bax was detected by Western blotting(WB).Results:Laser confocal microscopy showed that the immunofluorescence intensity of Bcl-2 protein gradually decreased with the increase of Ruanjian Sanjie Capsules(软坚散结胶囊)drug-containing serum concentration,and the fluorescence intensity of Bax protein gradually increased with the increase of dose concentration,and it entered the nucleus to varying degrees;WB detection showed that Bcl-2,Notch1,and NICD protein expression levels were inversely correlated with the drugcontaining serum dose concentration,that is,protein expression decreased with the increase of drugcontaining serum concentration,while Bax protein expression increased gradually with the increase of serum concentration.Afte 7860 cells knock out Notch1,the expression of related proteins was consistent with the expression trend of the drug-containing serum group.Conclusion:Ruanjian Sanjie Capsule's(软坚散结胶囊)low,medium and high dose concentration of drug-containing serum can promote 7860 cell apoptosis,and it is concentration-dependent.The molecular mechanism of apoptosis is directly related with Ruanjian Sanjie Capsule(软坚散结胶囊)blocking Notch signaling pathway activation.

Rupi Sanjie Xiaotong Plaster in Treating Breast Hyperplasia of Liver Stagnation and Phlegm Coagulation Pattern

Objective:To observe the clinical efcacy and safety of external application of Rupi Sanjie Xiaotong Plaster(乳癖散结消痛膏,RPSJXTP)in treating breast hyperplasia of liver stagnation and phlegm coagulation pattern.Methods:Totally 72 female patients with breast hyperplasia of liver stagnation and phlegm coagulation pattern admitted to the First Hospital Affiliated to Henan University of Chinese Medicine were enrolled by ran-domized,single-blind,placebo-controlled research method,and were divided into the treatment group(36 ca-ses)and the control group(36 cases)according to computer random number method.The treatment group were given RPSJXTP,while the control group were gi ven placebo plaster.The course of treatment in both groups was 8 weeks.The breast pain score,breast color ultrasound score,lump score,depression score,and Chinese medicine syndrome score were compared between the two groups at different times.The levels of estra-diol(E_(2)),progesterone(P)and pituitary prolactin(PRL)were detected and the adverse reactions of two groups were observed during the treatment of both groups.Results:Before treatment,there was no significant difference in total breast pain score,breast color ultrasound score,lump score,depression score and levels of serum P,E_(2)and PRL between the two groups(P>0.05);compared with previous treatment,the total breast pain score,breast color ultrasound score,lump score,and depression score significantly decreased in the treat-ment group(P<0.05),and the total breast pain score and depression score of the control group significantly declined(P<0.05);compared with control group,the total breast pain score,breast color ultrasound score,lump score,and depression score significantly decreased after treatment in the treatment group(P<0.05).The improvement rate of Chinese medicine syndromes and the clinical efective rate in the treatment group were higher than those in the control group(P<0.05).There were no serious systemie adverse reactions during the treatment in both groups.Local pruritus occurred in 3 cases in the treatment group and 1 case in the control group.Conclusion:RPSJXTP is effective in treating breast hyperplasia of liver stagnation and phlegm coagula-tion pattern.It can effectively relieve the breast pain which will not recur after stopping of drug use,which can reduce the color ultrasound score,lump score,and depression score of the patients and improve Chinese medi-cine syndromes without serious adverse reactions.

Efficacy and Safety of Sanjie Analgesic Capsule in Patients with Endometriosis-Associated Pain:A Multicenter,3:1 Randomized,Double-Blind,Placebo-Controlled Trial

Objective:To assess the efficacy and safety of Sanjie Analgesic Capsule(SAC)in Chinese patients with endometriosis-associated pain.Methods:This was a multicenter,randomized,double-blind,placebo-controlled trial conducted at 15 centers between November 2013 and July 2017 in China.Eligible 323 patients with endometriosis were randomized at a 3:1 ratio to the SAC group(241 cases)and placebo group(82 cases)by stratified block randomization.Patients in the SAC or placebo groups were given SAC or placebo 1.6 g 3 times per day,orally,respectively since the first day of menstruation for 3 consecutive menstrual cycles.The primary endpoint was clinical response to dysmenorrhea evaluated using a 10-point Visual Analogue Scale at 3 and 6 months.The secondary endpoint was the pain score evaluated by VAS(chronic pelvic pain,defecation pain,and dyspareunia)at 3 and 6 months,and the pain recurrence rate at 6 months.Adverse events(AEs)were recorded during the study.Results:A total of 241 women were included in the SAC group,and 82 were in the placebo group.Among these women,217(90.0%)and 71(86.6%)completed the intervention,respectively.At 3 months,overall response rate(ORR)was significantly higher in women administered SAC(80.1%)compared with those who received a placebo(30.5%,P<0.01).Six months after treatment,the ORR for dysmenorrhea was 62.7%in the SAC group and 31.7%in the placebo group(P<0.01).Chronic pelvic pain and defecation pain were significantly improved by SAC compared with placebo(both P<0.05).The incidence rates of total AEs events in the SAC and placebo groups were 6.6%and 9.8%,respectively,and no significant difference was shown between the two groups(P=0.339).Conclusion:SAC is well-tolerated and may improve dysmenorrhea in women with endometriosis-associated pain.

Xiaotan Sanjie recipe,a compound Chinese herbal medicine,inhibits gastric cancer metastasis by regulating GnT-V-mediated E-cadherin glycosylation

Objective:Xiaotan Sanjie recipe(XTSJ),a Chinese herbal compound medicine,exerts a significant inhibitory effect on gastric cancer(GC)metastasis.This work investigated the mechanism underlying the XTSJmediated inhibition of GC metastasis.Methods:The effect of XTSJ on GC metastasis and the associated mechanism were investigated in vitro,using GC cell lines,and in vivo,using a GC mouse model,by focusing on the expression of Glc-N-Actransferase V(GnT-V;encoded by MGAT5).Results:The migration and invasion ability of GC cells decreased significantly after XTSJ administration,which confirmed the efficacy of XTSJ in treating GC in vitro.XTSJ increased the accumulation of Ecadherin at junctions between GC cells,which was reversed by MGAT5 overexpression.XTSJ administration and MGAT5 knockdown alleviated the structural abnormality of the cell–cell junctions,while MGAT5 overexpression had the opposite effect.MGAT5 knockdown and XTSJ treatment also significantly increased the accumulation of proteins associated with the E-cadherin-mediated adherens junction complex.Furthermore,the expression of MGAT5 was significantly lower in the lungs of BGC-823-MGAT5+XTSJ mice than in those of BGC-823-MGAT5+solvent mice,indicating that the ability of gastric tumors to metastasize to the lung was decreased in vivo following XTSJ treatment.Conclusion:XTSJ prevented GC metastasis by inhibiting the GnT-V-mediated E-cadherin glycosylation and promoting the E-cadherin accumulation at cell–cell junctions.

Mechanism underlying efficacy of Shugan Sanjie decoction(疏肝散结汤) on plasma cell mastitis,based on network pharmacology and experimental verification

OBJECTIVE:To investigate the mechanism underpinning the effeicay of Shugan Sanjie decoction(疏肝散结汤,SGSJD)on plasma cell mastitis(PCM)based on network pharmacology,and to verify it through in vitro.METHODS:Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and Bioinformatics Analysis Tool for Molecular mechanism of Traditional Chinese Medicine were used to screen effective compounds and drug targets;Online Mendelian Inheritance in Man and Gene Cards were used to search for PCM targets.The potential targets of SGSJD in treating PCM were obtained after the drug targets and disease targets were crossed.Cytoscape software was used to establish and analyze the network of Chinese medicines-active compounds-targets-disease;STRING database platform was used to analyze Protein Protein Interaction network;Bioconductor software package was used to perform Gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment for potential targets.Western blot analysis was used to verify the janus kinase/signal transducer and activator of transcription(JAK-STAT)pathway in vitro.RESULTS:(a)47 potential pharmacological components of SGSJD treatment of PCM were screened including quercetin,luteolin,kaempferol and others;20 common targets were obtained,including interleukin-6(IL-6),epidermal growth factor receptor,estrogen receptor 1,nitric oxide synthase 3 and others;a number of signal pathways were available,of which advanced glycation end product/receptor for advanced glycation end products signaling pathway in diabetic complications,hypoxia-inducible factor 1 signaling pathway and janus tyrosine kinase-signal transducer and transcription activator(JAK-STAT)signaling pathway were the main signal pathways related to PCM.(b)Compared with the Blank group,the expressions of p-JAK2/JAK2,pSTAT3/STAT3 and IL-6 protein in the Model group were significantly increased(P<0.01);Compared with the Model group,the expression of p-JAK2/JAK2,p-STAT3/STAT3,and IL-6 protein in the treatment group were significantly reduced in a dose-dependent manner(P<0.05).Compared with the Model group,the dexamethasone significantly reduced the expression of p-JAK2/JAK2,p-STAT3/STAT3,and IL-6(P<0.01).CONCLUSIONS:The SGSJD may regulate the JAKSTAT signaling pathway to achieve the effect of treating PCM by reducing the expression of p-JAK2/JAK2,p-STAT3/STAT3 and IL-6 in a dose-dependent manner.