1.Carry out the study and application of tumor marker based on advanced scientific administration and technique
Chinese Journal of Laboratory Medicine 2001;0(04):-
The tumor marker(TM) is widely applied in early screening, thus providing further diagnostic hint of cancer, guiding the choice of therapy protocols for various tumor and evaluating the prognosis and therapeutic effects. However, there are some problems in TM application:There has still been no satisfactory early determinant TM, none test method of TM has been standardized, and the diagnostic capability of TM has not been evaluated reasonably. It is necessary to depend on the advanced scientific administration and technique, cooperate with clinical and basic disciplines actively, try to find out the new satisfactory TM and improve as well as exert important roles in the prevention, early diagnosis and treatment of 8 main catepories of cancers in our country.
2.Alteration of serum Cystatin C concentrations in patients with hepatopathy infected with hepatitis B or C virus
Jiafu FENG ; Tingmei CHEN ; Zhiguang TU
Chinese Journal of Laboratory Medicine 2003;0(11):-
Objective To investigate the clinical application of Cystatin C as a biological marker for monitoring hepatic pathological change in patients with virus hepatitis.Methods Two hundred and seven patients infected with hepatitis B or C virus(HBV, HCV)were divided into cirrhosis group(group A),chronic HBV group(group B),chronic HCV group(group C),and liver cancer group(group D). 32 healthy controls(group H) were recruited . The serum TIMP-1,TIMP-2,and Cystatin C as well as some traditional markers for monitoring liver function and renal function including creatinine, creatinine clearance rate, alanine transaminase, and aspartate transaminase were determined.Results In these groups, serum Cystatin C(F=28.334, P
3.Effects of quercetin on proliferation of lung cancer cell line A549 by down-regulating hTERT gene expression
Jian WANG ; Penghui ZHANG ; Zhiguang TU
Journal of Third Military Medical University 2003;0(19):-
Objective To investigate the effects of quercetin on the growth of lung cancer cell line A549 and the expression of hTERT gene. Methods The number of viable cells was ascertained by trypan blue dye exclusion test. Morphological changes of apoptotic cells were observed by electronic microscopy and DNA ladder assay. The telomerase activity was analyzed by PCR-TRAP assay and hTERT mRNA expression was detected by quantitative RT-PCR. Results Quercetin had a significant inhibition on the proliferation of A549 cells in a dose-dependent manner. The IC50 was 22.5 ?mol/L after exposure to quercetin for 48 h. The results from electron microscopy and DNA ladder showed that apoptosis occurred in the A549 cells of treatment groups. The results of quantitative RT-PCR and PCR-TRAP revealed that the expression of hTERT mRNA was significantly inhibited by quercetin and telomerase activity was decreased. Conclusion Quercetin inhibits the growth of lung cancer cell line A549 in a dose-dependent manner,and induce their apoptosis. The down-regulated expression of hTERT,suppression of telomerase activity and destruction of telomere stability may all contribute to the mechanism of apoptosis induction.
4.Analysis of tumor DNA in plasma for diagnosis of lung cancer
Xiaomei LUO ; Zhiguang TU ; Yulai WANG
Chinese Journal of Clinical Laboratory Science 2006;0(01):-
Objective To investigate the loss of heterozygosity (LOH) of 3p D3S1300 and D3S1289 loci of plasma DNA of lung cancer (LC) patients and evaluate its value for a tumor marker of lung cancer.Methods Two microsatellite markers,D3S1300 and D3S1289,were analyzed by PCR and silver staining to investigate LOH of the plasma DNA and cancer tissue DNA from 69 cases of primary lung cancer,and the plasma DNA from 40 control subjects.Results The percentages of D3S1300 LOH detected in the tumor tissue and plasma DNA of LC patients were 40.6% and 29.0% respectively,while the percentages of D3S1289 LOH were 31.9% and 24.6% respectively.In contrast,only 2 cases of D3S1300 LOH and 3 cases of D3S1289 LOH were found in plasma DNA of control group (P
5.The functions of GPⅠa/Ⅱa and GPⅣin platelet adhesion to collagen
Zhuo ZHANG ; Yulong CONG ; Zhiguang TU
Chinese Journal of Clinical Laboratory Science 2006;0(03):-
Objective To study the functions of GPⅠa/Ⅱa and GPⅣduring the course of platelet adhesion to collagen. Methods Reaction system which contains anti-GPⅠa/Ⅱa antibody ,anti-GPⅣantibody or no blocking antibody was analyzed by flow cytomety. PE labeled anti-CD42b antibody was used to select platelet and fluorescence intension of fluorescein isothiocyanate ( FITC) was detected during the course of platelet adhesion to FITC-labeled collagen. Results The antibody against GPⅠa/Ⅱa inhibited platelet adhesion to collagen, especially to activated platelet(P 0. 05 ). Conclusion GPⅠa/Ⅱa plays an important roles in platelet adhesion to collagen. Blocking GPⅠa/Ⅱa may decrease the adhesion. GPⅣmay be helpful for acceleration of platelet adhesion to collagen at early stage.
6.Function of GPⅠa/Ⅱa in the platelet-collagen interactions
Zhuo ZHANG ; Yulong CONG ; Zhiguang TU
Chinese Journal of Laboratory Medicine 2001;0(05):-
Objective To investigate the function of GPⅠa/Ⅱa in the platelet-collage adhesion and the platelet granules release induced by collagen. Methods With the flow cytometry, in either presence or absence of the anti-GPⅠa/Ⅱa antibody: (1) PE-labeled anti-CD42b antibody was used for selecting platelet and FITC fluorescence intension was detected during the course of platelet adhered to FITC-labeled collagen. (2) PerCP-labeled anti-CD61 antibody was used for selecting platelet and detecting CD62P with PE-labeled CD62P antibody during platelet granule release induced by collagen. Results Anti-GPⅠa/Ⅱa antibody inhibited the platelet-collagen adhesion, especially the activated platelet-collagen adhesion (P0.05). Conclusion GPⅠa/Ⅱa plays an important role in platelet-collagen adhesion. Blocking GPⅠa/Ⅱa can decrease the platelet-collagen adhesion, but has no effects on platelet granule release reaction.
7.Preparation,characterization and preliminary application of monoclonal antibody against cystatin C
Tingmei CHEN ; Jiafu FENG ; Zhiguang TU
Chinese Journal of Immunology 2000;0(09):-
Objective:To prepare monoclonal antibodies(McAb)against cystatin C(Cys C)and to establish the particle enhanced turbidimetric immunoassay(PETIA)for determining human serum Cys C.Methods:The prokaryotic expression vector pET32a(+)/Cys C was constructed and Cys C expression was induced.McAbs against Cys C were prepared with the hybridoma technique after mice were immunized with the purified recombinant protein.Then the McAbs were covalently attached to uniform microparticles,PETIA method for determination of human serum Cys C was established,and primary evaluation tests of methodology were performed.Results:Three hybridoma cell lines were obtained successfully,the secreted antibodies were isotype of IgG1,and Western blot confirmed that the antibodies reacted specifically to the Cys C protein.After one of the hybridoma cell lines was injected into mice abdominal cavity,the ascites abundant for McAb was obtained.The titer of the McAb against the purified protein was 1∶4?106.With the self-made McAb,PETIA for human serum Cys C was established.The primary evaluation tests of methodology revealed that self-established PETIA method had a satisfactory performance,which was equal to the import kit.Conclusion:The prepared McAb against Cys C is prepared,which could be used to establish PETIA for determining human serum Cys C.
8.Experimental study on shRNA targeted hTERT gene to suppress bladder cancer cell growth
Penghui ZHANG ; Lin ZOU ; Zhiguang TU
Chinese Journal of Urology 2001;0(04):-
Objective To explore the mechanism of siRNA targeted hTERT gene to inhibit bladder cancer T24 cell growth by decreasing c-myc and TGF-?1 expression. Methods shRNA-hTERT-pTZU6+1 vectors were constructed by RNAi-DNA vector technique, then the vectors were transfected into bladder cancer T24 cells,and the most effective vector and its optimal concentration were screened using RT-PCR to detect hTERT expression in T24 cells.The T24 cell growth, the alternative of cell phase,the expression of hTERT,c-myc and TGF-?1 were detected by flow cytometry,RT-PCR and immunohistochemistry. Results Three shRNA-hTERT-pTZU6+1 vectors were successfully constructed.The most effective vector was ph2-shRNA vector,and its optimal concentration was 1.0 ?g.This vector decreased the cell growth and the cell number of S phase from 65.2% to 38.6%,increased the cell number of G0/G1 phase from 32.0% to 57.9%,and attenuated both mRNA and protein expressions of hTERT,c-myc and TGF-?1 in T24 cells. Conclusions Targeted hTERT gene with siRNA may inhibit the cell proliferation of bladder cancer;down-regulating hTERT expression by attenuating the expression of c-myc and TGF-?1 is probably involved in the mechanism.
9.Regulation of hTERT promoter transcription activity by bHLH family genes
Penghui ZHANG ; Lin ZOU ; Zhiguang TU
Chinese Journal of Pathophysiology 1986;0(01):-
AIM: To investigate the transcription regulation of the promoter of human telomerase reverse transcriptase (hTERT) by transcription factors c-myc and [STBX]mad1. METHODS: The various plasmids including wild type hTERT (Tw) or mutant type hTERT (Td) which both harboring luciferase gene, the expression plasmids of c-myc and [STBX]mad1, and their control vectors were constructed. The plasmids were co-trans fected into bladder cancer cell lines T24, EJ and control cells COS-7 or fibrocytes by DOTAP liposome in various combining manner, respectively. The reporter gene luciferase activities in various groups were measured 48 h after transfection. RESULTS: The luciferase activities in T24 and EJ cells treated with Tw were much higher than that in COS-7 and fibrocytes cells treated with Tw, as well as higher than that in T24 and EJ cells treated with Td, respectively. In bladder cancer T24 and EJ cells, transcription factor c-myc and [STBX]mad1 positively and negatively regulated Tw expression in a dose-dependent manner. However, the effects of c-myc and [STBX]mad1 on Td were completely opposite to Tw. Combined with c-myc and [STBX]mad1, down-regulation of Tw expression was observed. CONCLUSION: c-myc and [STBX]mad1 regulates the transcriptional activity of hTERT promoter in bladder cancer cells, and the effects might highly depend on the conservative E-box sequence CACGTG.
10.THE PHARMACOKINETICS AND BILIARY EXCRETION OF PRAZIQUANTEL IN RATS BY DIFFERENT ROUTE OF ADMINISTRATION
Yong HUANG ; Yuzhu QUAN ; Zhiguang TU
Chinese Pharmacological Bulletin 1987;0(03):-
Praziquantel ( PQT ) concentrations in plasma after iv 20 mg/kg decayed rapidly with tip of 0.36 h. The absorption of PQT was rapid following the intramuscular doses of 10,20,40mg/kg or intragastic dose of 100mg/kg, but the phase of elimination was much longer than that after iv. Both of MAT1m and MATig were greater than MRTiv. The bioavailability of ig was 13.2%, suggesting a strong first-pass effect. The kinetics of PQT elimination was linear after intramuscular dose of either 10 or 20 mg/kg, but nonlinear process was found when the dose was increased to 40mg/kg.By any route of iv, im and ig administration, the concentrations of PQT in the bile were much lower than the peripheral plasma concentrations and changed in parallel to the later with high levels after iv, medium levels after im and much low levels after ig.