Expression levels of TGF-β1 mRNA in renal cortex of control group, diabetic group and taurine group measured by real-time quantitative RT-PCR were (7.0±0.8)×10 -3, (64.4±8.0)×10 -3 and (16.7±2.0)×10 -3, respectively. The corresponding results with end-point RT-PCR method were 0.28±0.12,0.58±0.16 and 0.43±0.10, respectively. Compared with end-point RT-PCR method, real-time quantitative with SYBR Green I was easier, faster, sensitive and specific.

objective To quantify the expression of kallikrein gene 6 (KLK6) in malignant and benign breast tissues and to statistically analyze the relationship between KLK6 expression levels and the clinico-pathological variables in patients with breast cancer. Method Paired breast tissue samples of cancerous and corresponding non-cancerous tissues were obtained from 48 patients with breast cancer who underwent surgical resection. Quantitative analysis of KLK6 mRNA expression was done using real-time quantitative reverse transcription-PCR (RT-PCR). The relationship between KLK6 expression intensity and various clinico-pathological variables was analyzed. In addition, the technique of small interfering RNA-mediated gene silencing was used to suppress the KLK6 expression in MCF-7, and to investigate the effects of KLK6 on cell proliferation rate and cell cycle. Results KLK6 mRNA over expression was found in cancerous tissues in 35 out of 48 patients (73%) as compared with normal breast tissue. The mean expression level of KLK6 mRNA in cancerous tissues was significantly higher than that in non-cancerous tissues (P

The human tissue kallikrein(KLK)gene family consists of 15 highly conservative serine pro-teases,which is the largest uninterrupted cluster of protease genes in the human genome. Several members of the family are expected to be markers for tumor diagnosis and prognosis. Studies find that the expressions of KLK2-11 and KLK13-15 are abnormal,and the majority of KLKs have potential diagnostic and prognostic values.

Objective To investigate the relationship between herpes simplex virus(HSV) and spontaneous abortion. Methods 40 aborted material and 16 post-aborted curettings were tested for the presence of HSVDNA with in situ hybridization.In addition 20 normal endometria and 20 induced abortive placentas were used as controls. Results 56.3 %(9/16)post-abortion curettings were demonstrated the presence of HSVDNA.It was higher than that of the control group( P

Objective To observe the clinical significance of detection of platelet-associated immunoglobulin (PAIgG).Methods Flow cytometry was used to detect PAIgG in 42 idiopathic throbocytopenic purpura (ITP) patients, 39 aplastic anemia (AA) patients, 21 patients with other autoimmune diseases and 20 normal controls. Rate of inefficient platelet infusion was calculated.Results Percentage of PAIgG positive in ITP, AA and other autoimmune diseases was higher than that in normal controls (P

Objective To investigate the significance of IL-18R? expression on CD4+ T cells in peripheral blood (PB) of children with Mycoplasma pneumoniae pneumonia (MPP).Methods T cell subtype CD3/CD4/CD8 and expression of IL-18R? on CD4+ T cells in PB from 35 children with MPP and 15 age- and sex-matched control subjects was determined by flow cytometry.Results Compared with that of healthy control, CD3+ and CD4+ positive cells in MPP children were decreased (P0.05).Conclusion There exists cell-mediated immune function disorders and Th1/Th2 imbalance in children with MPP. Th1 immune response is dominant in acute-stage MPP.

Objective To establish a rapid diagnostic method for the detection of marine vibrios,and then construct a new technology platform for the clinical diagnosis of marine vibrio infection.Methods A pair of PCR primers and a sequencing primer based on the whA gene of V.vulnificus and the toxR genes of V.parahemolyticus and V.alginolyticus were designed respectively,and then the specific DNA fragments were amplified.Next,the single-stranded DNA templates were prepared for pyrosequencing.The obtained base sequence was validated by NCBI alignment.In addition,the 16S rRNA genotyping of V.vulnificus was also performed.Results The PCR primers and sequencing primer of V.vulnificus showed good specificity,and a 167 bp DNA fragment was amplified from 4 strains of V.vulnificus.The pyrosequencing results completely matched with the whA gene sequence of V.vulnificus.Meanwhile,the control strains were negative.A 105 bp DNA fragment and a 134 bp DNA fragment were amplified from 11 strains of V.parahemolyticus and V.alginolyticus,respectively,and the pyrosequencing results were consistent with the expected sequence.In addition,one of 4 strains of V.vulnificus was identified as 16S rRNA-A type,and the other 3 as 16S rRNA-B type.Conclusion The PCR-pyrosequencing method established in this study is a new method for the real-time detection of short nucleotide sequences.It has some advantages such as high throughput,high precision and simple operation,and may be applied to the fast and accurate identification of marine and terrestrial pathogenic bacteria.

Objective To develop a real-time quantitative PCR method for detection of human breast cancer related novel gene-kallikrein gene 6 (klk6) expression and investigate klk6 expression levels in breast cancer tissue.Methods Using Sybr Green I, with GAPDH as reference, a real-time quantitative PCR method was established. klk6 expression levels of 32 normal and breast cancer tissues were detected and analyzed by the method and REST software.Results The amplification efficiencies for GAPDH and klk6 of the real-time quantitative PCR method were 0.90 And 0.95, respectively; inter-coefficient of variation were 1.0%~2.1% and 0.8%~1.2%,respectively; intra-coefficient of variation were 3.2% and 3.9%, respectively. The relative expression levels of klk6 in normal breast and breast cancer tissues were 0.017?0.009 and 0.040?0.017 with GAPDH as reference. Analysis results with REST indicated klk6 expression was up- regulated in breast cancer.Conclusion The real-time quantitative PCR method with Sybr Green I for klk6 mRNA quantification was simple, specific, reproductive and reliable, and could be used to study relationship betweeen klk6 expression and tumor.

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.

Objective The purpose of the current study was to develop a real-time quantitative reverse transcription-PCR method for detection of human kallikrein gene 5(KLK5) mRNA expression,quantify the expression of KLK5 mRNA in malignant and benign breast tissues,and statistically analyze whether KLK5 expression levels correlate with clinicopathologic variables in the patients with breast cancer.Methods Paired breast tissue samples from cancerous and corresponding noncancerous tissues were obtained from 48 patients with breast cancer who underwent surgical resection.The relationship of KLK5 mRNA expression status with various clinicopathologic variables were quantitativly analyzed by real-time RT-PCR.Results KLK5 mRNA expression in cancerous tissues was decreased in 38 of 48(79%) breast cancer patients compared with normal counterparts.The mean expression level of KLK5 mRNA in cancerous tissues was significantly lower than that in noncancerous tissues(P0.05).Conclusion The results of this study indicated that KLK5 mRNA expression was significantly lower in cancerous tissues than that in noncancerous breast tissues,and low expression of KLK5 mRNA correlated with TMN stage,metastasis and ER status.