1.Effect of free fatty acids on expression and secretion of extracellular matrix in rat mesangial cells
Dong LIANG ; Haiqing LUO ; Xiaowen CHEN ; Deshen TANG ; Huafeng LIU
Chinese Journal of Laboratory Medicine 2003;0(11):-
Objective To investigate the effects of Free fatty acids (FFAs) on extracellular matrix(ECM) mRNA expression and secretion.Methods Rat glomerular mesangial cells(HBZY-1 cells) were cultured in vitro and stimulated with OA in different concentration.The expression of collagen Ⅳ(Col Ⅳ) and fibronectin (FN) and transforming growth factor-beta1(TGF-?_1) mRNA were measured by semi-quantitative RT-PCR.The levels of Col Ⅳ and FN in cultured supernatant were determined by ELISA.Results The mRNA expression of Col Ⅳ, FN and TGF-?_1 of 25、100、400 ?mol/L stimulated OA groups were 0.94?0.17、1.16?0.15、1.28?0.19 and 0.82?0.11、0.97?0.07、1.09?0.08 and 1.15? 0.07、1.24?0.06、1.36?0.05 respectively , which increased significantly compared with their control group(0.73?0.16、0.53?0.09、 0.96?0.11 P
2.Clinical application of enzyme-linked immunospot assay for rapid diagnosis of active tuberculosis
Jun YUE ; Li LIANG ; Suhui LI ; Yuejian ZHANG ; Heping XIAO
Chinese Journal of Laboratory Medicine 2003;0(11):-
Objective To evaluate the clinical application of enzyme-linked immunospot(ELISPOT) assay for detection of M.tuberculosis antigen specific T cells in the rapid diagnosis of active tuberculosis.Methods Using the rapid enzyme-linked immunospot assay for detection of T cells with secretion of IFN-? specific for M.tuberculosis antigens in blood samples from 112 patients with tuberculosis, 24 patients with hepatitis C, 30 healthy persons, and 65 with other respiratory diseases.Results 107 of 112 tuberculosis patients had detectable M. tuberculosis antigen-specific T cells, whereas 2 of 30 healthy subjects and 65 patients with non-tuberculosis illnesses responded. This assay has a sensitivity of 95.5%, specificity of 93.3%.Conclusions M.tuberculosis specific T cells could serve as accurate markers of M.tuberculosis infection in an area of high tuberculosis prevalence.ELISPOT is a rapid, sensitive and specific method for detecting M.tuberculosis specific T cells.It also gives objective evidence to the diagnosis of active tuberculosis.
3.The study on the time resolved fluoroimmunoassay of double labeled prostate specific antigen
Lan ZHU ; Biao HUANG ; Xiangrui ZHANG
Chinese Journal of Laboratory Medicine 2003;0(11):-
Objective To establish the time-resolved fluoroimmunoassay (TRFIA) of total prostate specific antigen (tPSA) labeled with Sm 3+ and free prostate specific antigen (fPSA) labeled with Eu 3+ . Methods The monoclonal antibody (McAb) 101# was coated on the 96 well plates, anti-tPSA McAb 102# was labeled with Sm 3+ and anti-fPSA McAb 201# was labeled with Eu 3+ .The standard curves were given out by Log-Log_B function of the DEFIA instrument.The levels of PSA in sera from patients or healthy volunteers were determined by t/fPSA TRFIA using the autoDELFIA_ 1235 system.Results The measurement ranges of tPSA-TRFIA were 0.02-250 ?g/L and those of fPSA-TRFIA were 0.05-250 ?g/L. The within-run and between-run CVs of the tPSA-TRFIA were 2.38% and 3.91%, respectively, and those of fPSA-TRFIA were 3.16% and 3.34%, respectively. The recovery rates of tPSA-TRFIA and fPSA-TRFIA were 101.3% and 103.2%, respectively.The detection limitations of tPSA and fPSA were 0.05 ?g/L and 0.02 ?g/L,respectively.The average results of health group benign prostate disease patients,prostate cancer patients for tPSA were 1.4 ?g/L,4.3 ?g/L,14.2 ?g/L,and those of fPSA were 0.3 ?g/L,1.5 ?g/L,6.2 ?g/L.The cut-off point condition for benign prostate disease and malignant prostate cancer was tPSA
4.The clinical significance of epitopes of SmB and SmD in systemic lupus erythemotosus
Jing XUE ; Yang GAO ; Huaxiang WU ; Mengxue YU
Chinese Journal of Laboratory Medicine 2003;0(11):-
Objectives To assess the presence of autoantibodies directed against the epitopes of SmB and SmD in systemic lupus erythemotosus(SLE) as well as other different connective tissue diseases(CTDs) and analyze the clinical significance of them.Methods Polypeptides of SmB and SmD were designed with the animo acids sequence by the biologic technical software.The sera from different patients including 102 SLE,153 other CTDs and 54 controls were detected by ELISA with thesynthetic polypeptides.Results The positive percentage of anti-SmB and antiSm-D epitopes were higher in SLE than in other groups (P
5.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
6.Researching on the relation between lamivudine-resistant mutation and HBV genotypes
Junqiang LI ; Lijun LIU ; Feng LIU ; Shaocai DU
Chinese Journal of Laboratory Medicine 2003;0(11):-
Objective To analyze the relation between lamivudine-resistant mutation types and HBV genotypes.Methods 95 cases with YMDD mutation were selected from out-patient clinic and sickroom in our hospital.Restriction endonuclease MboⅠand EarⅠwere used to identify HBV genotypes of PCR product, farther a phylogenesis tree was applied to check it.Results We could divide 95 cases into two parts, 75 cases(78.95%) were HBV C genotype and 20 cases(21.05%) were B genotype, these were verified by phylogenesis tree.With regard to the types of YVDD mutation、YIDD mutation and YMDD+YVDD+YIDD mutation, there were 11 cases, 5 cases, 4 cases in genotype B, respectively, but 35 cases, 27 cases, 13 cases in genotype C.We made the cha-test to check genotypes and types of YMDD mutation, ?2= 0.856, P=0.710. There were no significant differences.Conclusions Using MboⅠand EarⅠcan genotype HBV PCR product easily and reliably. It is no statistical significance between HBV B or C genotype and the types of YMDD mutation.
7.Strengthen advancement of tumor tendency and earlier diagnostic technology
Chinese Journal of Laboratory Medicine 2003;0(11):-
The latest advancement on tumor biology and tumorigenesis has been discussed in this paper. The potential clinical significance on tumor tendency and early diagnosis of the genomic and proteomic analysis technology has been expounded too. It will be benefit for the new tumor biomarker studies.
8.Emphasis on the utility of laboratory examination on early diagnosis of chronic kidney disease
Chinese Journal of Laboratory Medicine 2003;0(11):-
Chronic kidney disease ( CKD) has become a pandemic.Laboratory assay takes the important role in early diagnosis of CKD.We should develop these assays correctly on the basis of full understanding of CKD and guide clinical use for them in laboratory medicine.
9.Mid-youth professionals:mainstay of laboratory medicine development
Chinese Journal of Laboratory Medicine 2003;0(12):-
The fifth mid-youth laboratory medicine congress was held by the Chinese Society of Laboratory Medicine and the Chinese Journal of Laboratory Medicine at Guiyang. The purpose of the congress was establish the communication platform for the development of mid-youth professionals. Regarding research development trend and problems of laboratory medicine, we prove some proposals for better development of mid-youth professionals: (1)Pay more attention to practice and ability; (2)Change concept, optimize orientation; (3)The research work should target “clinical practice”.
10.Application of ANAs protein chip in autoantibodies profile detection
Yongzhe LI ; Zhixian ZHAO ; Dawei TONG ; Shulan ZHANG ; Chaojun HU ; Yang GAO ; Weiping YANG ; Mengxue YU ; Liping ZHU ; Jing CHENG
Chinese Journal of Laboratory Medicine 2003;0(12):-
Objective To establish and apply the protein chip to detect eleven autoantibodies profile, and evaluate the authenticity and reliability with ANAs protein chip in clinical autoantibodies profile detection.Methods By comparing the results of IIF and ELISA , validation the sensitivity and specificity of ANAs protein chip in clinical autoantibodies profile detection. The autoantibodies detected were anti-SSA-52,anti-SSA-60,anti-SSB,anti-Sm,anti-RNP,anti-Scl-70,anti-Jo-1,anti-dsDNA,anti-rRNP,anti-centromere antibodies and antinuclear antibodies (ANA). To each autoantibody, we have selected 70 positive and 294 negative samples except the 32 rare samples that contain anti-Jo-1 antibody.Results The sensitivity to all the autoantibodies was 100% except anti-SSA52 and anti-SSB antibodies was 95.7%and 98.6% respectively. The specificity to all the autoanbodies was 100% except anti-SSB, anti-RNP-68, anti-Scl-70, anti-dsDNA, anti-CENP-B and ANA was 98.0%, 98.0%, 99.7%, 99.7%, 99.7% and 98.3% respectively. Conclusions To all the eleven antinuclear autoantibodies , the sensitivity is all above 95.0% and specificity is all above 98.0%, which indicate that there is high concordances between the ANAs protein chip and the methods used in clinical screening and confirmation,and it could meet the requirement of clinical autoantibodies profile detection. The protein chip method is fast, easy for detection with the characteristic of high-throughput,high sensitivity and specificity,it is hence recommended to apply ANAs protein chip to detect autoantibodies profile in clinical detection.