Objective To establish a nested-real-time-PCR for the detection of IL-21R mRNA in peripheral blood mononuclear cells(PBMC)of patients with rheumatoid arthritis,and explore its clinical significance.Methods The outer primers of span intron and inner primers were designed for nested-real-time-PCR.To increase specific template,the concentration of primers and enzyme,cycle times were decreased in the first amplification,and the real-time fluorescent quantity assay was performed in the second amplification.The ?-actin gene was used as internal reference,and the results were presented as the ratios of IL-21R mRNA to ?-actin mRNA.Results IL-21R mRNA was detected in 60 patients with RA and 30 healthy controls.The expression of IL-21R mRNA gene in the patients with RA was increased,and the statistical analysis has significant differences(P0.05),but a significant difference was found in the patients of grade III compared with those of grade I and II(P

Objectives To explore cholesteryl ester transfer protein(CETP)level in male patients with different degrees of coronary artery lesion and its clinical significance.Methods ELISA was used to measure CETP of 42 male patients with coronary heart disease(CHD)and 49 healthy male controls.The patients of CHD groups were subdivided into mono-vessel,ambi-vessel,multi-vessel lesion groups;localized,diffuse lesion groups;and mild,severe stenosis groups according to coronary angiography.Results The CETP level of patients with CHD(1.37?1.07 mg/L)was significantly higher than that of healthy control(0.99?0.53 mg/L)(P

Objective To research the relationship between the gene polymorphism of serum amyloid A protein(SAA)1 and coronary heart disease(CHD).Methods By using PCR-RFLP and sequencing,the gene polymorphism of SAA1 of 183 patients with coronary heart disease and 152 healthy controls were analyzed.Result In the both groups 3 alleles(1.1,1.3,1.5)and 6 genotypes(1.1/1.1,1.1/1.5,1.1/1.3,1.3/1.3,1.3/1.5 and 1.5/1.5)were found.The frequency of 1.5 allele in healthy controls group was notably higher than that in CHD group(P

Objective To investigate the expression of protease activated receptors(PARs)in mast cells.Methods Reverse transcription polymerase chain reaction(RT-PCR),flowcytometry and immunofluorescent cell staining were used to detect the expression of PARs in mast cell lines P815 and MC/9 at the levels of protein and mRNA.Results Both the P815 and MC/9 of mast cell lines expressed PAR-1,PAR-2,PAR-3 and PAR-4 at either protein or mRNA level.Conclusion The expression of all the four PARs in mast cells were detectable,which may be of significance for the further study on the function of PARs in mast cells.

Objective To develop a nested-real-time fluorescence quantitative PCR method for detection of TGF-?RII mRNA in human peripheral blood mononuclear cells(PBMC),and analyze the clinical implications of TGF-? RII in RA patients.Method The outer primers of span intron and inner primers were designed for nested-real-time fluorescence quantitative PCR.To increase specific template,the concentration of primers and enzyme,cycle times in the first amplification were decreased,and the real-time fluorescent quantity assay was performed in the second amplification.The ?-actin gene was used as internal reference,and the results were presented as the ratios of TGF-? RII mRNA to ?-actin mRNA.Results The standard curve showed a fine linear relationship between Ct(cycle threshold)and logarithm of template concentration,and the correlation coefficient was 0.999.The sensitivity reached 101 copies.The TGF-? RII gene expression in PBMC of RA patient was increased compared to healthy controls[(0.45?0.11)vs(0.37?0.10),P

Objective To investigate the predictive values of hemostatic activity of platelets on prophylactic transfusion,and determine the threshold of prophylactic platelet transfusion to avoid the bleeding risk caused by thrombocytopenia.Methods One hundred and twenty-seven patients whose platelet count(Plt)

Objective To investigate the correlation between the connective tissue growth factor(CTGF)in serum and the stages of liver fibrosis of patients with chronic liver disease,and explore new marker for evaluation of liver fibrosis.Methods The serum levels of CTGF in 313 patients with liver disease was detected by ELISA.Hyaluronic acid,type III procollagen,type IV collagen and laminin in serum were determined by RIA.Liver biopsy was performed in 45 patients with chronic liver disease.The quantitative relationship between the levels of CTGF and the stages of liver fibrosis was statistically analyzed by SPSS 11.5 software.Results A positive correlation between the serum levels of CTGF and the severity degree of chronic liver disease was found and the correlation coefficient was strong(r=0.634,P

Objective To study the relationship between drug resistance and expression of the gene associated with drug resistance in Candida albicans,as well as the secretory proteinase activity of these isolates.Methods The minimum inhibitory concentrations(MIC)to fouconzole and itraconazole in 99 strains of Candida albicans isolated from respiratory tract were measured by broth microdilution method,the M27-A protocol recommended by NCCLS.The gene expressions of CDR1 and MDR1 were semi-quantitatively determined by RT-RCR.The secretory proteinase activity of these isolates was assayed by plate with medium containing bovine serum albumin.Results The rates of drug resistance to fluconzole and itraconazole were 4% and 12.1% respectively.CDR1 expression in the susceptible dose-dependent(S-DD)isolates was higher than those in the susceptible isolates and drug-resistant isolates(P

Objective To evaluate the application of fluorescent multiplex amplification of short tandem repeat(STR)for monitoring survival of engraftment after bone marrow transplantation(BMT).Methods Three STR loci named D12S391,D18S865 and D20S161 in 56 cases were detected by fluorescent multiplex amplification.PCR products were separated and typed by DNA Sequencer.Results The genotypes of STR in 52 recipient after bone marrow transplantation were completely identical with those of the donors.In another 4 cases the evidences of mixed chimerism were observed.Conclusion The system of fluorescent multiplex amplification of STRs exhibited high capacity of discrimination and low cost.Its application in the detection of STR after BMT is reliable,sensitive and simple.Combined with the clinical manifestation it can be used to evaluate the effect of BMT.

Objective To develop a real-time fluorescence quantitative PCR(FQ-RCR)method for detection of Kallkreins(KLK)4 gene expression in human breast cancer,and to investigate KLK4 expression levels in breast cancer and normal tissues.Methods KLK4 expression levels of 25 normal breast tissues and 39 cancer tissues were detected by the developed real-time quantitative PCR method.The statistical analysis for the relationship between KLK4 expression and several pathological parameters was performed by t test.Results The levels of KLK4 mRNA in normal breast and breast cancer tissue were 0.0120?0.0044 and 0.0272?0.0067 respectively(P0.05).Conclusions The level of KLK4 mRNA in breast cancer tissue was higher significantly than that in normal breast tissue.The results indicated that KLK4 gene expression may have relevance with breast cancer development but have no significant relevance with ER,PR,CerbB-2 and tumor metastasis.