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.Research progress on induction of pluripotent stem cell from adult stem cell
Suwen QI ; Yong DAI ; Zhiguang TU
International Journal of Biomedical Engineering 2011;34(3):188-192
It is confirmed that the adult cells can be re-programmed to embryonic stem cells(ESCs) by presenting some certain factors in oocytes in the clone process of animals. In recent years, some transcription factors that can induce pluripotent stem cells(iPS) have been identified and which made it possible to obtain induced pluripotent stem cells similar to embryonic stem cells. iPS provides a unique platform to study the pluripotent mechanism and to establish some specific disease models. This major scientific discovery can not only avoid the use of ES which involves ethics debate, but also lead the stem cell research to a new field.
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.Elevated adiponectin: a danger signal for type 1 diabetic patients
Yiting TU ; Xia LI ; Zhiguang ZHOU
Chinese Journal of Endocrinology and Metabolism 2014;30(4):288-291
Serum adiponectin levels in type 1 diabetic patients are higher than those in the normal population.We reviewed the influencing factors of serum adiponectin levels in type 1 diabetic patients,the relationship between adiponectin and adverse type 1 diabetic clinical features,complications,concurrent condition,and all-cause mortality,as well as potential mechanisms and research directions.
5.Study of the enzyme assay of dimethylarginine dimethylaminohydrolase in rabbit kidney
Jian QIN ; Yunzhen CHEN ; Zhiguang TU ;
Chinese Pharmacological Bulletin 1987;0(03):-
AIM To establish the method for the enzyme assay of dimethylarginine dimethylaminohydrolase (DDAH) in rabbit kidney and to determine the optimal condition for the assaying. METHODS Five healthy Japanese rabbits weighing 3 0 to 3 5 Kg were killed by air embolism,kidneys were harvested and then were homogenized. Asymmetric dimethylarginine (ADMA) was used as the substrate for DDAH. UV 265 spectrophotometer was applied to determine the amount of the enzymatic product L Citrulline( L Cit).The amount of L Cit produced under different conditions was compared and the optimal condition was screened. The kinetic parameters of ADMA degraded by DDAH were calculated. RESULTS The kinetic parameters of ADMA degraded by DDAH were as follows: K m=(0 28?0 10) mmol?L -1 , V m=(1 36?0 42) mmol?L -1 ?g -1 ?min -1 . The optimal conditions for the enzyme assay of DDAH in rabbit kidney determined in this study follow: The concentration of the enzyme protein was 12 g?L -1 ,the optimal pH of the buffer was 7 4,the final concentration of ADMA was 2 mmol?L -1 ,and the reaction time was 30 min. CONCLUSION The method offered here is easily done. The concentration of the substrate determined in this study was based on the value of Km,thus it was beneficial to the accurate assay of DDAH.
6.Effect of matrine on PLA_2 activity of LPS-induced inflammatory rats and its mechanism
Geng QIU ; Zhiguang TU ; Xiaowen LI ;
Chinese Traditional and Herbal Drugs 1994;0(07):-
Object To study the antiinflammatory effect of matrine and its active mechanism. Methods The matrine effects on activities of secretory phospholipase A 2 (sPLA 2) in peripheral serum and cytosolic phospholipase A 2 (cPLA 2) in leucocyte were measured with Escherichia coli membrane incorporated by arachidonic acid as the substrate; the Ca 2+ level of leucocyte was determined using Frua2 AM loading method, and the rat plantar swelling test was used to examine the antiinflammatory effect of matrine. Results One hour after ip 30 mg/mL matrine, the inhibitory rate of sPLA 2 activity was (81.9?1.8)% , cPLA 2 (28.4?6.0) %, while Ca 2+ concentration was (157.10?20.56) nmol/L which was 15.3% higher than that of the control. Plantar swelling test showed that matrine had a significant anti inflammatory effect. Conclusion Matrine is a novel PLA 2 inhibitor with anti inflammatory effect.
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.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.
9.Screening of serum biomarkers and establishment of decision tree in silica-exposed populations by SELDI-TOF-MS
Xuefeng ZHAO ; Shixin WANG ; Zhiguang TU
International Journal of Laboratory Medicine 2008;29(6):492-495,封3
Objective To screen the serum biomarkers by surface-enhanced laser desorption-ionization time-of-flight mass spectrometry (SELDI-TOF-MS) and establish the decision-tree model for the silica-exposed populations.Methods In addition to 20 healthy people without silica-exposed history chosen as control group,all the subjects diagnosed as different stages (stage 0 group,silica-exposed population,n=20;stage 0+ group, doubtful silicosis,n=20;stage Ⅰ group,stage Ⅰ silicosis) of silicosis according to the national diagnostic criteria (GBZ70-2002) of pneumonoconiosis with no complications were recruited. SELDI-TOF-MS and CM10 chip assay were applied to acquiring serum protein finger printing. The bioinformatic software was adopted to perform data ananlysis and establish Wilks'lambda decision equation and decision tree.Results As compared with that of control group, differential peaks appeared in three groups of silica-exposed population, accounting for 80% of total differential peaks. Wilks'lambda decision equation was established with M/Z 3711.73, M/Z 4183.66, M/Z 5487.28 and M/Z 6124.64, which sensitivity and specificity in differentiating healthy controls and silica-exposed population was 95% and 90% respectively. The sensitivity and specificity of decision tree was 100% and 95% respectively. Neither single peak nor Wilks'lambda decision equation worked well for discrimination of silica-exposed population, while the discriminating rates of decision tree accounted for 100%, 95%, 85% and 90% for healthy control group, stage 0 group, stage 0+ group, stage Ⅰ group respectively.Conclusion M/Z 5933.63,M/Z 4183.66,M/Z 5487.28,M/Z 6124.64,M/Z 5915.14,M/Z 2745.7, M/Z 3711.73, M/Z 2954.78 and M/Z 3401.03 are potential biomarkers for discriminating healthy controls and silica-exposed population, and the decision-tree works well for discrimination of controls, silica-exposed population, doubtful silicosis patients and stage Ⅰ silicosis ones.
10.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.