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.

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 investigates the gene expression of telomerase by real-time quantitative telomeric-repeat amplification protocol assay (RTQ-TRAP) in the peripheral blood mononuclear cells (PBMC) of colorectal carcinoma patients and the relationship between the telomerase gene expression in PBMC and clinicopathological features. Methods Peripheral blood samptes were collected from 71 colorectal carcinoma patients, 20 benign colorectal disease patients and 25 normal controls. The telomerase gene expression in PBMC was measured by RTQ-TRAP, and serum CEA in colorectal carcinoma patients was measured by chemiluminescence immunoassay. Results The gene expression of telomerase in PBMC was positive in 50 out of 71 cancer patients (70. 4% ) , 1 out of benign patients (5. 0% ), respectively. The difference of the telomerase gene expression of PBMC in benign colorectal diseases and cancer patients was significant (χ2 = 24. 521, P < 0. 001 ). There was no significant association between the expression of telomerase and patient's gender, age, Dukes stage, and tumor site. The positive rate of CEA in colorectal carcinoma patients was not significantly higher than positive rate of telomerase gene expression(χ2 = 2. 286,P = 0. 125). Conclusions The RTQ-TRAP method is highly accurate and sensitive in measuring telomerase gene expression. The detection of telomerase gene expression in PBMC of colorectal carcinoma patients is a simple and useful molecular marker for the diagnosis of colorectal carcinoma.

Objective To explore the peptides that can mimic the blood type A antigen and evaluate the anti-A antibody detection value of these peptides.Methods The anti-A monoclonal antibodv (NaM87-1F6)was used to panning the phage clones from a phage display 12-mer peptide library.Positive clones were identified by phage ELISA,phage mieropanning methods.Phage DNA Was sequenced and the corresponding peptide sequences were deduced.Agglutination inhibition test WaS performed to assess the ability of phage clones to inhibit the binding between the type A red blood cell and the anti-A antibody. ABO-ELISA based on the selected peptides was compared with classical haemagglutination test jn the detection of senlm anti-A antibody.Results Seven positive clones were chosen after panning,phage ELISA and phage micropanning.Six clones displayed peptide EYWYCGMNRTGC(C5),the other one displayed peptide QIWYERTLPFTF(C17).The phages displaying the selected peptides could specifically inhibit agglutination of type A red blood cells(RBCs)by anti-A antibodies.In the ABO-ELISA based on C5 and C17,the receiver operating characteristic(ROC)Curve showed that area under curve(AUC)were 0.889 (P=0.000),0.75l(P=0.000)respectively.The Spearman correlation Coeffieient between the ABO-EliSA value and the antibody titer derived from haemagglutination assay were 0.743(P<0.01),0.664(P<0.01)respectively.As for C5,0.300 was the best cut-off for ABO-ELISA with 82.2% sensitivity and 83.3% specificity.As for C17,the sensitivity and specificity of ABO-ELISA was 68.9% and 63.3% respectively when the cut-off value was 0.250.Conclusions The peptides EYWYCGMNRTGC and QIWYERTLPFTF can mimic the blood type A antigenic epitope.ABO-ELISA based on these peptides has the potential for the detection of anti-A antibody.

Objective To revise the Chinese version of the National Institutes of Health-Chronic Prostatitis Symptom Index (CHN-NIH-CPSD), and evaluate its feasibility, reliability, validity and responsiveness. Methods The NIH-CPSI was translated into Chinese according to a standard methodology including forward-backward-forward technique. The CHN-NIH-CPSI was pre-tested in consecutive samples of 162 native-speaking Chinese chronic prostatitis(CP)patients. Ninety-five of 162 filled the index again on the same day and after 4-week therapy. Ninety-seven healthy men were included as evaluated. Results The recovery of the questionnaires was 100% and all the patients filled the index completely. The mean time to complete the questionnaire for the patient group was 5.2±2.4 (range 2 - 12) min. The split-half reliability was 0.82. For the overall index and each subscale, the test-retest reliability was 0.98, 0. 98, 0. 98, 0. 97, respectively(P＜0.01);and the Cronbach's α coefficient was 0. 61,0. 71, 0. 59, 0. 75, respectively. The confirmatory factor analysis showed good construct validity with a goodness of fit index of 0. 85 and a x2 of 124.67(P＜0. 01). Of all 162 patients, the scores of the overall index and each subscale were 23. 33±5.91. 8. 80±4.26, 5.30±2.82, 9. 23±1.90, respectively;and those of healthy controls were 1. 95±1.97, 0. 37±1.03, 0. 15±0.58, 1.42± 1.20,respectively. Of the 95 patients, the original scores were 23. 53±5.60, 9.21 ±4.04, 5.10±2.75,9.21 ±2.05, comparing with 19.47±6.36, 7.79±3.95, 3. 58±1.88, 8.11±2.50, the 4 weeks later scores. The group t-test and paired t-test showed good responsiveness. Conclusions The CHN-NIH-CPSI has high feasibility, reliability, validity and responsiveness for testing the patients with CP. It is suitable for Chinese-speaking patients and helpful for cross-cultural comparisons of men with CP in clinical and research settings.

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.

AIM: To observe whether transfection of mammalian expression vector pEGFP containing the gene of B-cell specific moloney leukemia virus insertion site 1(BMI-1) could express in human cervix cancer cell line HeLa, and to detect its effect on HOX family expression and cell cycle.METHODS: pEGFP-BMI-1 was transfected into HeLa cells with Lipofectamine 2000. The expression of pEGFP-BMI-1 was determined by EGFP fluorescence and Western blotting. SYBR green I real-time RT-PCR was used to quantitate mRNA expression of P16~(INK4a), hTERT, HOXA9, HOXB4 and HOXC13. FACS analysis was used to detect the change of cell cycle.RESULTS: In HeLa cells transfected with pEGFP-BMI-1, the results of real-time RT-PCR showed that the mRNA expressions of P16~(INK4a), HOXA9 and HOXC13 were reduced to 9.2%, 10.9% and 69.7%, respectively, as compared to control HeLa cells (P<0.01). However, hTERT and HOXB4 mRNA expressions did not change significantly (P>0.05). FACS analysis showed a decrease from 65.68 % to 50.53% in G_1 population and a significant increase from 27.17% to 39.59 % in S population after transfection (P<0.01).CONCLUSION: BMI-1 over-expression in HeLa cells down-regulates mRNA expressions of P16~(INK4a), HOXA9 and HOXC13, decreases G_1 population and increases S population. Therefore, BMI-1 may be involved in carcinogenesis and cancer development.

ObjectiveTo optimize Chonglian oral solution extracting craft. MethodsWith the obtaining rate of extract and the total content of the Pariphyllin Ⅰ and Phillyrin presented in the extract as the indexes for the water extraction process,and with the total content of the Pariphyllin Ⅰ and Phillyrin presented in the extract as the indexes for the alcohol deposition process,orthogonal design was used to optimize the conditions for the extraction process respectively. ResultsThe optimal conditions for the water extraction of Chonglian oral solution was as following:to add water 10 times,decocting 3 times,1 hour each time.The optimal conditions for the alcohol deposition of Chonglian oral solution was as following:concentrated for the relative density to 1.10(80C hot test),cold,add ethanol to the solution for the ethanol content of the solution reached 80%,static settlement for 24 hours. ConclusionThe extracting method is reasonable,stable,and suitable to industrialized producting.