Objective To investigate the correlation between Toll-like receptor2 (TLR2) gene promoter region -597T/C polymorphism and primary ANCA associated small vasculitis (AAV) in Guangxi Han people. Methods A case contrastive control study was adopted in the study. Patients with AAV (patients group, n=110) and healthy people (control group, n = 200) were recruited. Associated serum indexes were detected and polymorphisms of TLR2 gene promoter 597T/C were analyzed by polymerase chain restricted fragment length polymorphisms (PCR-RFLP). Results (1)Three TLR2-597T/C genotypes were discovered in 110 AAV patients, namely, TT, TC and CC, with the frequency of 54.55%,40.00% and 5.45% respectively. And the frequencies of allele T and C were 74.55% and 25.45%. In control group, the genotype frequencies of TT, TC and CC were 56.00%,40.50% and 3.50%, with 76.25% of allele T and 23.75% of allele C. No significant differences were found in neither genotype distribution nor allele frequencies between the patients group and control group ( P > 0 . 05 ) . ( 2 ) Significant differences were found in the incidence of proteinuria rate and the hemoglobin (P< 0.05)in AAV patients. (3)There was no significant difference between AI and CI in TT, TC and CC genotype in AAV patients. Conclusions Polymorphism of TLR2-597T/C may be correlated with the incidence of proteinuria and the level of hemoglobin, while no obvious correlation with the genetic susceptibility of ANCA in vasculitis patients of Guangxi Han people.

Objective To investigate the correlation between toll-like receptor 9 (TLR9) gene 2848G/A polymorphism and primary antineutrophil cytoplasmic antibodies (ANCA) associated small vasculitis (AAV). Methods A case-control study was performed among 135 patients diagnosed with AAV and 140 disease-free control and we test the serum biochemical parameter. Polymorphism was analyzed by polymerase chain restricted fragments length polymorphism. As for statistic method, according to the character of data, we performed t-test, chi-square test, Spearman grade related analysis and one-way ANOVA. Results ① The frequencies of AA, GG, GA genotype of TLR9 2848 in AAV patients were 14.07%, 38.52%, and 47.71%, respectively; ② Significant increase in IgM was observed in AA genotype than GG+GA genotype in AAV patients (F=4.561, P<0.05). ③ There was no significant difference between AI and CI in AA, GA and GG genotype in AAV patients (F=2.115, 0.760, P>0.05). Conclusion AA, GA and GG genotypes are detected in TLR9 2848G/A in patients with AAV in Guangxi, without significant correlation with susceptibility to primary AAV in Guangxi.

Objective: To evaluate whether human NK cells expanded in vitro can be used as carrier cells of reovirus and to investigate its clinical application value. Methods: Expansion of human NK cells in vitro, and flow cytometry was used to analyse the purity of CD3-CD56+ cells. Expanded NK cells were loaded with reovirus and observed by confocal microscopy, to determining the location of reovirus on NK cells. CCK-8 assay was used to detect reovirus-induced oncolysis of expanded NK cells carrying reovirus (Reo-NK) to tumor cells in the presence of neutralizing antibodies; Real-time fluorescence quantitative PCR was used to assess the relative expression of viral RNA in tumor cells. Cytotoxicity assay were performed to detect Reo-NK cells against KRAS mutant (DLD-1) and KRAS wild type (CaCo-2, HT29) colorectal cancer cell lines, ELISA matched paired antibodies assay was performed to measure the perforin level released by NK cells. Results: Confocal microscopy demonstrated that NK cells retained reovirus on the surface. Expanded NK cells could delivery reovirus to tumor cells in the presence of neutralizing antibodies, and the reovirus after delivery still had significant oncolytic activity (P<0.01); Corresponding qPCR result displayed that the expression of viral RNA in tumor cells significantly increased over time (P<0.01). Compared with NK group, Reo-NK group evidently enhanced the cytotoxicity on colorectal cancer cell lines with both KRAS gene mutant and wild (all P<0.05), and significantly increased the release of perforin (all P<0.05). Conclusion: In vitro expanded NK cells provide a convincing cell carrier for reovirus, while reovirus enhances the cytotoxicity of NK cells, and the combination of the two show a stronger killing effect on colorectal cancer cells，that has important clinical application value.