Objective:To set up a real-time fluorescent RT-PCR and to quantitate the expression levels of hTERT mRNA in peripheral blood mononuclear cells from patients with AML and to observe the correlation between the expression of hTERT mRNA and occurrence and relapse of AML and to probe into the correlation between the expression of hTERT mRNA and telomerase activity.Methods:Real time fluorescent RT-PCR and Lightcycler PCR system were used to quantitate expression levels of hTERT mRNA.PCR-ELISA was used to quantitate telomerase activity.Results:①N_(hTERT) from AML at initial presention,AML at relapse,AML at complete remission and health examinee was 299.2?292.8,550.1?441.3,14.0?9.2 and 12.3?6.7 respectively and the expression levels of hTERT mRNA from AML at initial presention and AML at relapse elevated signficantly comparing to that from AML at complete remission and health examinee,moreover the expression levels from AML at relapse was higher than that from AML at initial presention significantly.②The telomerase activity from AML at initial presention,AML at relapse,AML at complete remission and health examinee was 32.8%?24.3%,48.6%?31.4%,7.4%?5.1% and 7.6%?3.6% respectively and the telomerase activity from AML at initial presention and AML at relapse elevated signficantly comparing to that from AML at complete remission and health examinee,moreover telomerase activity from AML at relapse was higher than that from AML at initial presention significantly.③There was a strong correlation between telomerase activity and the expression of hTERT mRNA and the correlation coeffecients was 0.78.Conclusion:The up-regulation of the expression levels of hTERT mRNA and telomerase activity are one of important factors during occurrence and relapse of AML,moreover there is a strong correlation between telomerase activity and the expression of hTERT mRNA.

Objective To set up a real-time quantitative assay method of insertion sequence 6110 DNA of mycobacterium tuberculosis, and explore its value for diagnosing tuberculosis. Methods Real-time quantitative assay of insertion sequence 6110 DNA of mycobacterium tuberculosis was performed with Taqman technique and Lightcycler quantitative PCR system. Results 213 clinical samples of different types were detected, 32 cases were positive, and the positive rate was 15 02%. The scope of quantitative results of positive samples was 3 1～7 2?10 6 copies/ml, and the sensitivity and specificity for diagnosis of tuberculosis were 82 76% and 95 65% respectively Conclusion Real-time quantitative assay of insertion sequence 6110 DNA of mycobacterium tuberculosis is a rapid and effective method for diagnosis of tuberculosis.

Objective To investigate the relations of the expression of bcl-xL in superficial bladder tumor and prognosis. Method The expression of bcl-xL was detected by envision system in 80 cases of superficial bladder tumor. Results In patients with high expression of bcl-xL, the recurrence rate was higher than that of normal expression [71.4%(30/42) vs 50.0%(19/38) ,P < 0.05], and the recurrence of time as early as normal expression [(16.0 ± 1.2) months vs (36.0 ± 4.5) months](P < 0.05). Conclusion The expression of bcl-xL could effectively predict the prognosis of patients with bladder tumor, those who with high expression of-bcl-xL have bad prognosis.

Objective To study whether ORF3 coded by fap1-orf4 gene locus of Streptococcus pa-rasanguis is involved in the regulation of Fap1 glycosylation and maturation and to investigate whether ORF3 influences Streptococcus parasanguis adhesion. Methods A gene replacement strategy was adapted to con-struct orf3 alleic replace mutant of Streptococcus parasanguis. Complementation assay and Western blot were used to test Fap1 expression levels. Whole saliva-coated hydroxyapatite (SHA) adhesion assay was adapted to determine Streptococcus parasanguis adhension. Results (1) Non-polar was found in strain VT1774, the orf3 alleic replace mutant of Streptococcus parasanguis. (2) Western blot showed that mature Fapl (Mr about 220 × 103) disappeared and were substituted with high molecular weight Fapl (Mr about 470 × 103) in strain VT1774, furthermore, complementation assay showed VT1775, the complementation strain of VT1774, re-stored mature Fapl expression. (3) The binding ability reduced significantly in strain VT1774. Conclusion ORF3 coded byfapl-orf4 gone locus was required for Fap1 glycosylation and maturation in Streptococcus pa-rasanguis, orf3 alleic replacment resulted in Fap1 glycosylation and mature disorder and decreasing of adhen-sion ability of Streptococcus parasanguis.

RNA interference(RNAi), a highly conserved process of post-transcriptional gene silencing, can be induced by small interfering double-stranded RNA that mediate sequence-specific mRNA degradation. In the past several years, RNAi has been widely used as both an experimental tool to study mammalian gene function and a potential therapeutic approach to treat human diseases. In addition, some new proteins which involve in RNAi pathway have been characterized in mammalian cells. Here, we summarize the various molecules in RNAi, mechanism of action, and the current therapeutic applications in cancers.

Objective To investigate whether two polymorphism sites of the the exon 2 Lys65Lys(G/A) and exon 9 Va1365Met(G/A) in T ceils immunoglobulin domain and mucin domain protein-4(TIM-4) are associated with asthma in Chinese Han population of Hubei province. Methods The polymorphisms were de-tected with polymerase ehain reaction-restriction fragment length polymorphism(PCR-RFLP) in 185 cases of al-lergic asthma and 162 healthy controls. The genotype and allele frequencies were calculated and analyzed. Re-sults(1)The genotype frequencies of G/G, G/A and A/A in Lys65Lys(G/A)polymorphism were 0.840, 0.160 and 0 respectively in the healthy population, and were 0.859, 0.141 and 0 respectively in the allergic asthma population. No significant difference in genotype and allele frequencies was found between asthma pa-tients and the control subjects (P=0.603, P=0.618). (2) The polymorphism of the Val365 Met(G/A) was not detected in our study. Conclusion There is polymorphism site of the exon 2 Lys65Lys(G/A)in TIM-4, but this polymorphism site is not associated with asthma in Han nationality in Hubei Chinese population. There is no SNP of the exon 9 Val365Met(G/A) in TIM-4 in Chinese Han population of Hubei province.

Objective To investigate whether glucosyltransfernse(GTF) and open reading frame 4 (ORF4) coded by fap1-orf4 gene locus of Streptococcus parasanguis was involved in the regulation of Fap1 glycosylation, and mature and to determine whether there was interaction between GTF and ORF4. Methods A gene replacement strategy was adapted to construct gtf and orf4 allele replace mutant of S. Parasanguis. Complementation assay and Western blot were used to test fap1 expression levels. Yeast two-hybrid analysis and GST pull down assays were adapted to determine the interaction between GTF and ORF4. Results (1) Compared with wild S. Parasanguis, mature Fapl (Mr about 220×103) disappeared and were substituted with high molecular weight Fapl (Mr about 360×103) in gtf or orf4 alleie replace mutants of S. Parasan-guis. Complementation assay showed that pVPT-GFP-gtf and pVPT-GFP-orf4 restored mature fap1 expression in gtf or orf4 alleie replace mutants, respectively. (2) With Yeast two-hybrid analysis, the eotransformants, AH109/pAD-Gtf+pBD-orf4 and AHlOg/pAD-orf4+pBD-gtf growed on SD-LTHA selective ngar plate after streaked, reversely, the eotransformants, AH109/pAD+pBD-orf4,AH109/pAD+pBD-gtf、AH109/pBD+ pAD-orf4、AH109/pBD+pAD-gtf did not grow on SD-LTHA selective agar plate, furthermore, the cotrans-formants, AH109/pAD+pBD-orf4 and AH109/pAD-orf4+pBD-gtfshowed blue during X-α-gal assay. (3) GST pull down assay confirmed the direct interaction between GTF and ORF4. Conclusion There is inter-action between GTF and ORF4 coded byfapl-orf4 gene locus of S. Parasangnis and the formation of the GTF and ORF4 complex was required for the glycosylation and mature of Fapl in S. Parasanguis.

Objective To investigate the clinical significance of human papillomavirus(HPV )genotype combined with thinprep cytologic test(TCT )in the diagnosis of cervical lesion .Methods A total of 473 patients were checked for 21 subtypes of HPV by diversion hybrid gene chip technology ,TCT and colposcope biopsy were also detected at the same time .The histology was selected as a gold standard to analyze the tested results .Results The over all positive rate of HPV in 473 patients was 35 .7% .The positive rate of high risk HPV(HR‐HPV) was 32 .1% ,the positive rate of TCT was 26 .6% ,the sensibility ,specificity ,false negative rate , positive predictive value and negative predictive value between HR‐HPV detection and TCT detection were no statistical signifi‐cance(χ2 = 3 .444 ,P= 0 .063) .The sensibility ,specificity ,false negative rate ,positive predictive value and negative predictive value of combining test were 95 .8% ,77 .7% ,4 .2% ,52 .3% and 98 .7% ,the sensibility and negative predictive value improved notably , and the omission diagnose rate decreased significantly(P< 0 .05) .Conclusion HPV genotype combined with TCT detection could significantly improve sensibility and negative predictive value and decrease omission diagnose rate in diagnosis of cervical lesion .

Telomerase activity and the expression of telomerase subunits (for example, telomerase reverse transcriptase and telomerase associated protein 1 and telomerase RNA component) of peripheral white blood cells were detected in the patients with acute myelogenous leukaemia (AML) and the correlation between telomerase activity and the expression of telomerase subunits was observed. In 94 peripheral white blood cells from 18 healthy volunteers and 76 patients with AML, including 31 AML at initial presentation, 24 at relapse and 21 at complete remission, the telomerase activity and telomerase subunits mRNA or RNA were detected by PCR-ELISA and RT-PCR respectively. The results showed that the positive rate of telomerase from patients with AML at initial presentation, at relapse and at complete remission was 74.1%, 79.2% and 4.8% respectively. The positive rate of telomerase reverse transcriptase mRNA from healthy volunteers, AML at initial presentation, AML at relapse and AML at complete remission was 5.6%, 80.6%, 83.3% and 9.5% respectively. The positive rate of telomerase associated protein 1 mRNA and telomerase RNA component in all samples were 100%. It was suggested that the up-regulation of telomerase activity and the expression of telomerase reverse transcriptase is correlated closely with the occurrence and relapse of AML, so telomerase activity and the expression of telomerase reverse transcriptase may be used to estimate the curative effect and predict relapse of AML. Moreover, the up-regulation of telomerase activity is correlated with the expression of telomerase reverse transcriptase significantly.
Carrier Proteins/*metabolism ; DNA-Binding Proteins ; Leukemia, Myelocytic, Acute/*enzymology ; RNA/metabolism ; RNA, Messenger/metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Telomerase/*metabolism

Objective To study the effect of Tacrolimus on blood lipid after renal transplantation,and the relationship between C825T polymorphism in G protein beta 3 subunit (GNB3) gene and serum lipid levels.Method Eighty-one cases of recipients patients after renal transplantation were divided into two groups in terms of Tacrolimus concentration:normal blood concentration group (group A) and low blood concentration group (group B).The serum lipid levels at 1st,3rd,6th,and 12th month after renal transplantation were measured.Genotype was determined by the simple sequence-specific primer polymerase chain reaction (SSP-PCR).Result The percentage of patients with hypertriglyceride in group A was significantly higher than in group B during the one-year follow-up period.There was significant difference between the two groups in the serum triglyceride levels but no difference in the serum cholesterol levels.The 825C/T polymorphism in the GNB3 gene was not associated with hypertriglyceride in renal transplantations in Wenzhou.Conclusion The serum triglyceride levels in renal transplantations in Wenzhou was associated with the Tacrolimus concentration,and the incidence of hypertriglyceride is not associated with the 825C/T polymorphism in the GNB3 gene.