Objective To investigate the role and molecular mechanisms of bladder cancer-derived exosomal long non-coding RNA(lncRNA)CIAT1 in mediating collective invasion and to evaluate its clinical significance and potential therapeutic value.Methods High-throughput sequencing was used to identify lncRNAs that are highly expressed in exosomes from bladder cancer and lymph node metastatic tissues.CIAT1 was selected for further validation in clinical bladder cancer samples.By constructing CIAT1-overexpressing and knockdown bladder cancer cells,we demonstrated in vitro that CIAT1-contained exosomes target cancer-associated fibroblasts(CAFs)to induce collective invasion.The underlying mechanism of CIAT1 in bladder cancer collective invasion was explored through RNA pull-down,RNA immunoprecipitation(RIP),dual-luciferase reporter assays,chromatin isolation by RNA purification(ChIRP)and chromatin immunoprecipitation(ChIP).Results CIAT1 was significantly upregulated in exosomes derived from bladder cancer tissues compared to adjacent normal tissues by High-throughput sequencing(fold change>1.5,P<0.05)and clinical sample validation(P<0.01).In vitro experiments,exosomal CIAT1 was selectively internalized by cancer-associated fibroblasts(CAFs),significantly enhancing collective invasion of bladder cancer via regulating CAFs.In co-culture models,CIAT1 overexpression group showed increased total number and total length of collective invasion chains compared to the control group(P<0.01 for both).Mechanistically,CIAT1 was packaged into exosomes via binding to hnRNPA2B1,and internalized by CAFs,where it activated N-cadherin transcription by modulating H3K4me3 histone modification at the N-cadherin promoter.Consistently,the CIAT1 overexpression group exhibited elevated collective invasion chain numbers and lengths compared to the control group(P<0.01 for both).However,blocking N-cadherin reversed the pro-invasive effects of CIAT1,with no significant differences in chain numbers or lengths between the CIAT1 overexpression+N-cadherin blockade group and controls(P>0.05 for both).Further clinical correlation analysis confirmed that CIAT1-regulated N-cadherin is closely associated with collective invasion in bladder cancer patients(P<0.01).Conclusions Exosomal CIAT1 derived from bladder cancer cells targets CAFs to activate N-cadherin transcription,thereby promoting bladder cancer collective invasion.

Objective To investigate the role of circular RNA,circMYCBP2,which is highly expressed in lymph node(LN)metastatic bladder cancer(BCa)tissues,in enhancing BCa cell adhesion and mediating lymphovascular invasion(LVI),and to evaluate its clinical relevance and potential therapeutic value.Methods High-throughput sequencing identified circRNAs with high expression in LN metastatic BCa tissues.Further validation of their expression in bladder cancer tissues was conducted through clinical samples.The coding ability of circMYCBP2 was predicted by circBank bioinformatics website,and the encoded short peptide was detected by immunoprecipitation(co-IP)combined with silver staining and Western blot(WB).Then,by constructing BCa cells with overexpression and knockdown of circMYCBP2 in vitro,the invasive ability of BCa cells was verified by wound healing and transendothelial cell migration assays.The underlying mechanism of circMYCBP2 in the forma-tion of LVI was explored through RNA pulldown,qRT-PCR and WB.Results High-throughput sequencing and clinical sample validation confirmed that circMYCBP2 is highly expressed in LN metastatic BCa.In vitro experi-ments demonstrated that circMYCBP2 overexpression significantly enhanced BCa cell migration across lymphatic endothelial cells.Mechanistically,circMYCBP2 encodes the short peptide MYCBP2-227aa via an IRES-dependent mechanism by interacting with EIF3H.Overexpression of MYCBP2-227aa increased the mRNA stability of VCAM-1,thereby enhancing the invasive capacity of UM-UC-3 cells.Conclusions CircMYCBP2 encodes the short peptide MYCBP2-227aa through an EIF3H-mediated,IRES-dependent translation mechanism.MYCBP2-227aa regulates VCAM-1 expression and promotes the invasive behavior of BCa cells.Our findings elucidate the critical biological role and molecular mechanism of circMYCBP2 in the formation of LVI and LN metastasis of BCa,providing a potential biomarker and therapeutic target for early lymphatic metastasis in BCa.

Objective To investigate the role and molecular mechanisms of bladder cancer-derived exosomal long non-coding RNA(lncRNA)CIAT1 in mediating collective invasion and to evaluate its clinical significance and potential therapeutic value.Methods High-throughput sequencing was used to identify lncRNAs that are highly expressed in exosomes from bladder cancer and lymph node metastatic tissues.CIAT1 was selected for further validation in clinical bladder cancer samples.By constructing CIAT1-overexpressing and knockdown bladder cancer cells,we demonstrated in vitro that CIAT1-contained exosomes target cancer-associated fibroblasts(CAFs)to induce collective invasion.The underlying mechanism of CIAT1 in bladder cancer collective invasion was explored through RNA pull-down,RNA immunoprecipitation(RIP),dual-luciferase reporter assays,chromatin isolation by RNA purification(ChIRP)and chromatin immunoprecipitation(ChIP).Results CIAT1 was significantly upregulated in exosomes derived from bladder cancer tissues compared to adjacent normal tissues by High-throughput sequencing(fold change>1.5,P<0.05)and clinical sample validation(P<0.01).In vitro experiments,exosomal CIAT1 was selectively internalized by cancer-associated fibroblasts(CAFs),significantly enhancing collective invasion of bladder cancer via regulating CAFs.In co-culture models,CIAT1 overexpression group showed increased total number and total length of collective invasion chains compared to the control group(P<0.01 for both).Mechanistically,CIAT1 was packaged into exosomes via binding to hnRNPA2B1,and internalized by CAFs,where it activated N-cadherin transcription by modulating H3K4me3 histone modification at the N-cadherin promoter.Consistently,the CIAT1 overexpression group exhibited elevated collective invasion chain numbers and lengths compared to the control group(P<0.01 for both).However,blocking N-cadherin reversed the pro-invasive effects of CIAT1,with no significant differences in chain numbers or lengths between the CIAT1 overexpression+N-cadherin blockade group and controls(P>0.05 for both).Further clinical correlation analysis confirmed that CIAT1-regulated N-cadherin is closely associated with collective invasion in bladder cancer patients(P<0.01).Conclusions Exosomal CIAT1 derived from bladder cancer cells targets CAFs to activate N-cadherin transcription,thereby promoting bladder cancer collective invasion.

Objective To investigate the role of circular RNA,circMYCBP2,which is highly expressed in lymph node(LN)metastatic bladder cancer(BCa)tissues,in enhancing BCa cell adhesion and mediating lymphovascular invasion(LVI),and to evaluate its clinical relevance and potential therapeutic value.Methods High-throughput sequencing identified circRNAs with high expression in LN metastatic BCa tissues.Further validation of their expression in bladder cancer tissues was conducted through clinical samples.The coding ability of circMYCBP2 was predicted by circBank bioinformatics website,and the encoded short peptide was detected by immunoprecipitation(co-IP)combined with silver staining and Western blot(WB).Then,by constructing BCa cells with overexpression and knockdown of circMYCBP2 in vitro,the invasive ability of BCa cells was verified by wound healing and transendothelial cell migration assays.The underlying mechanism of circMYCBP2 in the forma-tion of LVI was explored through RNA pulldown,qRT-PCR and WB.Results High-throughput sequencing and clinical sample validation confirmed that circMYCBP2 is highly expressed in LN metastatic BCa.In vitro experi-ments demonstrated that circMYCBP2 overexpression significantly enhanced BCa cell migration across lymphatic endothelial cells.Mechanistically,circMYCBP2 encodes the short peptide MYCBP2-227aa via an IRES-dependent mechanism by interacting with EIF3H.Overexpression of MYCBP2-227aa increased the mRNA stability of VCAM-1,thereby enhancing the invasive capacity of UM-UC-3 cells.Conclusions CircMYCBP2 encodes the short peptide MYCBP2-227aa through an EIF3H-mediated,IRES-dependent translation mechanism.MYCBP2-227aa regulates VCAM-1 expression and promotes the invasive behavior of BCa cells.Our findings elucidate the critical biological role and molecular mechanism of circMYCBP2 in the formation of LVI and LN metastasis of BCa,providing a potential biomarker and therapeutic target for early lymphatic metastasis in BCa.

OBJECTIVETo explore the anti-platelet activation effect and partial mechanisms of Taohong Siwu decoction (TSD).

METHODThe effect to venous thrombosis model and pulmonary thromboembolism model induced by vein injecting ADP and Adr was observed. The platelet adhesion rate was analyzed by using spinning glass bottle, and the platelet aggregation rate induced by ADP, Adr was analyzed by using turbidimetry. The acute blood stasis rat model was established to analyze the content of plasm TXB2 and PGI2 by RIA, and the content of VWF, GMP-14 by ELISA.

RESULTTSD could effectively reduce platelet the adhesion rate of normal rat, inhibit the platelet aggregation of normal rat induced by ADP, Adr. It significantly reduced the plasma TXB2 VWF, and GMP-140 level of blood stasis rats. It also had significant tendency to increase 6-keto-PGF1alpha level.

CONCLUSIONTSD possessed obvious activity of inhibiting platelet activation. The mechanism related with the restraining of platelet adhesion, platelet aggregation and platelet releasion.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Male ; Medicine, Chinese Traditional ; Platelet Activation ; drug effects ; Platelet Aggregation ; drug effects ; Rats

G-protein coupled receptors(GPCRs)belong to the biggest subfamily of transmembran receptors.Recently,more and more research has been suggested that the dimerization of GPCRs may regulate the physiological and pharmacological activity.With the development of biochemistry technology and molecular biology,a great progress has been achieved in the field of the dimerization of GPCRs.This article will generally demonstrate that the vital roles of the homodimerization and heterdimerization.

AIM:To study the isolation and identification of curdione from Zedoary oil. METHODS: Silica column was adopted to isolate constituents from Zedoary oil;IR,GC/MS and NMR methods were used to identify its structure. RESULTS: Three constituents were isolated from Zedoary oil,including crystal C_1,C_2 and C_3.Crystal C_1 was identified to be curdione and curdione content in Zedoary oil was 11.31%. CONCLUSION: This method is simple and convenient and it can be used to isolate more quantity of curdione for further pharmaceutical study.

AIM: To investigate the protective effects and mechanisms of Tongqiao Huoxue Decoction on the rats of cerebral ischemia. METHODS: Cerebral ischemia madel was established on the basis of cerebral thrombus and at the same time ligaturing bilateral carotid arteries.These models were used to observe the protective effects of Tongqiao Huoxue Decoction on contents of CGRP、ET、IL-1?,TNF? in rats′ cerebral ischemica. RESULTS: The results showed that Tongqiao Huoxue Decoction decreased the contents of IL-1?、TNF-?、 ET,and increased the contents of CGRP. CONCLUSION: Tongqiao Huoxue Decoction had effects on anti-cerebral ischemia.The protective effects of Tongqiao Huoxue Decoction are related to decreasing the contents of IL-1?,TNF-?,ET,and increasing the contents of CGRP.

The anti-inflammatory of waterand fat-soluble alkoloids of Leon-tice Kiagnanensis (WSALK, FSALK ) were described after sc of WSALK (150, 300 mg/kg ) and FSALK (13, 26mg/kg ) . The increased capillary permeability in abdominal cavity and skin of mice induced respectively by 0.7% HAc and histamin were inhibited abviously. WSALK (75, 150, 300mg/kg, ip ) and FSALK (6.5, 13, 26mg/Kg, ip ) markedly inhibited the ear inflammation induced by xylene in mice, carrageenin-iaduces swelling of the ankle in normal on adrenalecto-mized rats, and carrageenin-induced pleurisy in rats. Both compound also inhibited the granuloma induced by cotton pellet after sc of WSALK 300, 600 mg/kg and FSALK 13, 26 mg/kg qd ? 7d.Both compound decreased the content of PGE in exudate formed after injecting carrageenin into the hind paw of rats. These results suggest that both compounds passesses the anti-inflammatory activity and their actions might be relatied to their inhibiting the synthesis or relase of pro-staglandin E. WSALK TI 9.35, FSALK TI 4.50.

The effects of TSCP and TSCC on aggregation and adhesion of blood platelets were measured by turbidimetric method and salzman method respectively. TSCP and TSCC strongly increase mouse and rat platelet aggregation and platelet adhesion. These effects are important for blood coagulation. cAMP and TXB2 levels in plasma and platelet were determind by RIA TSCP significantly increased the TXB2 level in plasma and platelet. This effect may be the main mechanism of increasing platelet aggregation by TSCP.