1.The effects of mycophenolate mofetil on the expression of CD40L molecule by activated HUVEC
Chinese Journal of Immunology 2000;0(09):-
Objective:To investigate the effects of mycophenolate mofetil(MMF)on the expression of endothelial CD40L and lymphocyte-endothelium adhesion.Methods:HUVEC was used for adhesion assay and CD40L detection.The effect of mycophenolic acid(MPA),an active metabolite of MMF,was observed in the study.HUVEC were treated with TNF-? or/and MPA for 24 hours.The lymphocyte-endothelium adhesion was detected by Rose Bengal staining.After treated with TNF-?,rIFN-?,and LPS,HUVEC CD40L expression was detected.HUVEC was treated with any of the modulators or each combined with MPA,respectively for 24 hours and different doses of MPA combined with LPS were also designed to stimulate HUVEC for 24 hours.The expression of CD40L on HUVEC was detected by cell-ELISA method.Results:MPA inhibited adhesion to lymphocytes of the HUVECs of either resting or stimulated with TNF-?.The cytokines used in the study induced expression of CD40L on HUVECs.MPA did not inhibit CD40L expression on resitng HUVECs,whereas it inhitbited CD40L expression induced by TNF-?,rIFN-?,or LPS,and the inhibition of CD40L expression on the cells induced by LPS was shown in dose of MMF-dependnet pattern.Conclusion:Endothelial cells express a low level of constitutive CD40L that is increased by treated with simple cytokines.MMF inhibits interactions between endothelial cells and lymphocytes by suppressing the expression of CD4OL on endothelial cells.
2.Programmed Cell Death of Leukemic Cell(HL-60) Induced by Quercetin in Vitro
Chinese Traditional Patent Medicine 1992;0(10):-
To study anti-leukemic cell mechanism of quercetin, its effects on the proliferation activity of human leukemic cell (HL-60) in vitro were investigated by flow cytometric method and MTT assay.The studies indicated that quercetin showed strong suppression on the proliferation activity of leukmic cells in the time and dose-dependent manner. The value of IC50 and 95% confidence limit were 43. 1 (30. 5~60. 8)?mol/L after 48h. The flow cytometry indicated that it could induce programmed cell death of leukmia HL-60 ecll.
3.Immune active effects of chemokine RANTES on human peripheral mononuclear cells
Xiao GU ; Xiaoda TANG ; Shenyang GU
Chinese Journal of Organ Transplantation 1996;0(03):-
Objective To investigate the proliferation and phenotypes of peripheral mononuclear cells (PMNC) stimulated by recombinant human RANTES (rhRANTES) and the mechanisms involved.Methods PMNC was stimulated by various concentrations of rhRANTES and/or anti-CD3 mAb and intervened by PDTC and CTLA4Ig. Scintillation counter was used to count the cpm of proliferation cells and flow cytometry was used to detect the phenotypes of lymphocytes.Results rhRANTES was capable of directly stimulating purified human PMNC proliferation and two peaks occurred with rhRANTES concentration of 100 ng/ml and 5000 ng/ml respectively. The proliferation of PMNC stimulated by rhRNATES was significantly higher than that in the presence of anti-CD3mAb (P
4.Developments in chemotherapy for patients with advanced colorectal cancer
Journal of International Oncology 2006;0(09):-
Surgical resection has been accepted as a radical therapy for patients with colorectal cancer, but most of them will develop regional recurrence or distant metastasis. With the development and improvement of novel drugs and methods, they will bring survival hope to patients. In this article, we review the developments in chemotherapy for patients with advanced colorectal cancer.
5.Effects of cyclooxygenase-2 silencing by adenovirus-delivered siRNA on the relevant changes of human esophageal carcinoma cells
Hui ZHAO ; Xiao-Meng GU ;
Chinese Journal of Digestion 2001;0(12):-
Objective To investigate of human esophageal squamous carcinoma cells(Eca-109) in which cyclooxygenase (COX)-2 gene was knocked down by adenovirus delivered siRNA.Methods Based on the plasmid pSUPER cloned with RNA polymerase Ⅲ-dependent promoter HI,the interfering plasmid psiRNA/COX-2 targeting human COX-2 mRNA was constructed,The siRNA/COX-2 fragment was derived from psiRNA/COX-2 digested by Not I and Xho 1.and was cloned into the shuttle plasmid pAdTrack.Then pAdTrack/siRNA/COX-2 was obtained and co transfected into the E.coli strain BJ5183 with the bone plasmid pAdEasy-1,the recombinant adenovirus Ad/siRNA/COX-2 was generated by homologous recombination.Having been packaged and amplified in cells 293,Ad/siRNA/COX-2 was transfected into Eca-109 cells.The PGE2 concentration in the cells culture supernatant was determined by ELISA,and the level of COX-2 mRNA in the cells was tested by real time PCR.Moreover,cell cycle and apoptosis were determined by flow cytometry,and cells growth curve was protracted.Results The recombinant adenovirus Ad/siRNA/COX-2 was successfully constructed.Ninty six hours after Ad/ siRNA/COX 2 transfecting into Eca 109 cells,COX-2 mRNA was reduced by 71.7%,and PGE2 concentration in the cells culture supernatant was decreased by 62.0%.Correspondingly,the growth of cells slowed down.At the same time,the cells in G0-G1 phase was increased by 32.24%,and those in S phase and G2-M phase were reduced by 16.38% and 15.86%.respectively.And cells apoptosis index was increased by 9.19%.Conclusion The adenovirus based-RNAi was capable of knocking down remarkably COX-2 of human esopbageal carcinoma cells,which lead to growth of cells slowing down.
6.Expression and clinical implication of CC chemokines and receptor in long-term survived renal transplant recipients
Xiao GU ; Xiaoda TANG ; Jiaquan XIAO
Chinese Journal of Urology 2000;0(12):-
Objective To investigate the expression and clinical implication of CC chemokines and receptor in long surviving kidney grafted patients of different immune states. Methods 73 cases of recipients surviving more than 3 years were divided into three groups: control group of normal renal function(C group,n=32), group under low dosage of immunosuppressants(L group,n=20) and group with chronic allograft dysfounction(D group,n=21). The plasma RANTES、MCP 1 and MIP 1?were detected by sandwich ELISA.The expression of CCR5 was determined by flow cytometry(FACS). Results Concentrations of RANTES and MIP 1? as well as expression rate of CCR5 in L group were lower than those of C group ( P 0.05).Plasma level of MCP 1 was significantly higher in group D than that in group C( P 0.05). Conclusions Expression of CC chemokines and receptor closely correlated with the immune state of renal transplant recipients and could be valuable to estimate and monitor the immune state of patients.
7.Implication of expression of lymphotactin mRNA in allografts
Xiao GU ; Xiaoda TANG ; Shenyang GU ; Al ET
Chinese Journal of Organ Transplantation 1996;0(03):-
Objective To investigate the implication of lymphotactin (LTN) mRNA expression in cardiac allografts and the effect of cyclosporin A (CsA).Methods Three groups of rats underwent the heterotopic cardiac transplantation: isograft group (group A), untreated group (group B) and CsA treated group (group C). The LTN mRNA expression was detected by one step RT PCR method. Results The expression of LTN mRNA was not detected in both isografts and normal hearts. The changes of LTN mRNA expression were correlated with the process of acute allograft rejection. The peak of elevated level of the LTN mRNA expression appeared in the 5th day after transplantation and CsA could delay the peak and downregulate its expression. The peak level of LTN mRNA expression in group C was significantly lower than that in group B ( P
9.Effect of Biglycan and FAK signal pathway on the proliferation of colon cancer cells and its mechanisms
Xiaojing XING ; Xiaohu GU ; Jingdong XIAO
Practical Oncology Journal 2015;29(5):428-431
Objective To investigate the effect of Biglycan and FAK signal pathway on the proliferation of colon cancer cells in vitro and its possible mechanisms.Methods Biglycan expression vector was constructed and transfected into the colon cancer cell line HCT116.FAK inhibitor was used for cell treatment as well.Cells were divided into 5 groups:control group(HCT116),control group transfected with empty plasmid(Vector),con-trol group with empty plasmid transfected and inhibitor treatment(Vector+PF-562271),group transfected with Biglycan expression vector(Biglycan),group with Biglycan expression vector transfected and inhibitor treatment ( Biglycan+PF-562271) .Treatment duration was 24 hours.The expressions of FAK,p-FAK,PCNA and p53 were detected by Western Blot.The proliferation of cells was detected by MTT.Results The overexpression of Biglycan significantly promoted the proliferation of HCT116 and the phosphorylation of FAK(P<0.01).It signif-icantly up-regulated PCNA and down-regulated p53(P<0.01).The FAK inhibitor PF-562271 treatment could obviously inhibit the proliferation of HCT116,and the regulation of Biglycan on the expression of p-FAK, PCNA.p53 proteins was reversed(P<0.01).Conclusion Biglycan regulates the proliferation of colon cancer cells by promoting the activation of FAK signal pathway.
10.Effects and mechanism of Biglycan and FAK signaling pathway on the invasion and metastasis of colon cancer cells
Xiaohu GU ; Jingdong XIAO ; Xiaojing XING
Practical Oncology Journal 2015;29(5):444-449
Objective To observe the influence of Biglycan on the focal adhesion kinase( FAK) activa-tion and to explore whether it regulates the invasion and metastasis of colon cancer through FAK signaling path-way.Methods The overexpressive plasmid of Biglycan was constructed and then transfected into the colon canc-er cell line HCT116.Meanwhile,FAK inhibitor was used to treat cells.Control group(HCT116),empty plasmid group(Vector),empty plasmid and inhibitor treatment group(Vector +PF -562271),Biglycan overexpression group( Biglycan) ,Biglycan overexpression and inhibitor treatment group( Biglycan+PF-562271) were set.Twen-ty four hours after exposure to inhibitor,the expression of FAK and p-FAK in each group was detected by West-ern Blot.The invasion and metastasis of colon cancer cells was detected by transwell assay.Results Overexpres-sion of Biglycan significantly enhanced the phosphorylation level of FAK and promoted the invasion and metastasis of HCT116(P<0.01).The inhibitor of FAK PF-562271 could significantly reduce the expression of p-FAK and reverse the effect of Biglycan on the invasion and metastasis of colon cancer cells.Conclusion Biglycan reg-ulates the invasion and metastasis of colon cancer cells through activation FAK signaling pathway.