1.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.
2.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.
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
8.Application of ProtEx~(TM):an exogenous FasL protein modification technology in the study of immunomodulation
Xiao GU ; Jin YANG ; Hong ZHAO
Chinese Journal of Organ Transplantation 2010;31(3):144-147
Objective By using ProtEx~(TM) technology to decorate target cells with an exogenous protein and observe its function for immunomodulation.Methods Donor splenocytes were decorated with chimeric SA-FasL protein.The expression of FasL and its impact on other molecules on the surface of splenocytes was detected.Heterotopic rat heart transplantation was performed from inbred WF rat to ACI rat.According to the different donor splenocytes perioperatively injected into recipients,the recipients were divided into 3 groups:SA-FasL group (n=23),SA group (n=20) and control group(n=10).Heart beating was examined by palpation on daily basis,the cessation was regarded as reiection,while surival over 100 days was considered as graft long-term survival.Diabetes models in C57BL/6 mice were induced by intravenous injection of STZ.Islets from BALB/C mice were decorated by SA-FasL protein,islets transplant was performed from BABL/c to C57BL/6 mice.Based on the different treatments of donor islets,recipients were divided into three groups:islet-FasL group (n=21),islet-SA group (n=21) and islet-untreated group(n=14).Rejection was identified while urine glucose was positive and blood glucose value was over 250 mg/ml for consecutive two days.Urine glucose negative over 100 days was regarded as graft long-term survival.Results The mean exDression rate of FasL in splenocytes was 94.49%±4.27% tested by flow cytometry and FasL showed red color 0n the surface of splenocytes by fluorescent stammg.FasL deeoration did not disturb the expression of CD3,CD4,D8,MHC I and CD80.The long-term survival rate o,f heart transplant showed significant difference between tWO groups(SA-FasL:70% vs SA:25%,P<0.05).67% FasL-decorated islets had normal function 30 days post transplant,and 29%FasL-decorated islets achieved long-term survival,which was significantly differed from the other groups(P<0.05).Conclusion ProtEx~(TM) is a simple,fast,safe and efficient approach for exogenous protein modmcation.Decoration of target cells with Chimeric FasL by this technique plays an umportant role in immunomodulatiorn.
9.The effect of early application of Tirofiban on CRP levels in patients with acute myocardial infarction after emergency percutaneous coronary intervention
Hongbing XIAO ; Jun GU ; Dadong ZHANG
Journal of Interventional Radiology 2010;19(2):101-104
Objective To evaluate the effect of Tirofiban on CRP levels in patients with acute myocardial infarction (AMI) after primary emergency percutaneous coronary intervention (PCI). Methods Eighty-four AMI patients admitted on emergency were randomly divided into two groups: (1) early-treated group (n=45), immediately receiving Tirofiban intravenously on admission and (2) late-treated group (n=39), receiving Tirofiban intravenously after coronary angiography was performed. TIMI grading before and after PCI in beth groups were compared, CRP levels before and three days after PCI were estimated. The major adverse cardiovascular events (MACEs) occurred during hospitalization and following-up period of three months were recorded. Results Before PCI, TIMI grade 3 forward flow rate in early-treated group was significantly higher than that in late-treated group, while no significant difference existed between two groups after PCI. Three days after PCI, CRP level in early-treated group was markedly lower than that in late-treated group. During hospitalization, the occurrence of MACEs in early-treated group was lower than that in late-treated group, while no marked difference was found between two groups during the following-up period of three months. Conclusion In treating AMI patients with primary PCI, Tirofiban should be used as early as possible, which is safe and effective for PCI and can also significantly improve forward blood flow in target vessels, decrease the ClIP level and reduce the occurrence of MACEs during hospitalization.
10.Down-regulation of midkine by small interfering RNA increase chemosensitivity in bladder cancer cells
Yu FAN ; Xiao GU ; Xiaoda TANG
Chinese Journal of Urology 2010;31(9):604-607
Objective To study the effects of knock down midkine(MK)by siRNA on chemosinsitivity in bladder cancer cells. Methods Three MK siRNAs were designed and constructed. After transfected with MK siRNA or scrambled siRNA for different time, cultured cells were harvested to carry on the next experiments. Expression of MK was determined by real-time RT-PCR and western blot, and apoptosis were evaluated by caspase-3 activity and TUNEL assay. MTT was performed to examine the inhibition effect of Paclitaxel (PTX) on cells. Results MK siRNA could down-regulate the MK expression in a dose- and time-dependent manner. The MTT results showed that the inhibit rates were (18.21±0.36)%, (18.19±0.29)%, (17.89±0.33)%, (1.86±0.52)%, (32.56±0.53) %, (53. 83±0.38) % and (78. 95±0.55) % in different groups PTX alone(0.2μmol/L), ConA +PTX(0.2 μmol/L), Con-B +PTX(0.2 μmol/L), siRNA alone(12. 50 nmol/L), siRNA(3. 125nmol/L) + PTX (0. 2 μmol/L), siRNA (6. 25 nmol/L) + PTX (0. 2 μmol/L) and siRNA (12. 50nmol/L)+PTX(0.2 μmol/L), respectively. The TUNEL results showed that apoptosis index was (1.81 ±0. 36)%, (1. 89±0. 38)%, (5. 56±0. 58)%, (9. 68±0.55)% and (15. 25±0.56)% in different groups (Con-A, Con-B, siRNA (3. 125 nmol/L), siRNA (6. 25 nmol/L) and siRNA ( 12. 5nmol/L), respectively. The activity of caspase-3 increased significantly in transefected cells with a dose-and time-dependent manner. Conclusion MK siRNA could sensitize human bladder cancer cells to chemotherapy which might be through the apotosis.