1.Serological characteristics of HIV antibody indeterminant and the discrimination assay
Zuoyi BAO ; Xiaoli YANG ; Yongjian LIU ; Haishan GENG ; Lin LI ; Hanping LI ; Jingyun LI
Chinese Journal of Microbiology and Immunology 2010;30(5):427-430
Objective To clarify the serological characteristics and predictive value of HIV antibody indeterminant and to evaluate the efficiency of 3 assays to discriminate HIV antibody indeterminant.Methods Three hundred and ninety-four HIV antibody indeterminant serum samples were collected and the Western blot pattern were analyzed.Ninety-seven HIV antibody indeterminant individuals were followed up,and the development of HIV antibody were observed.The initial serum samples of 67 followed individuals were tested by viral load,line immunoblot assay and ELISA for HIV-1 p24,with the golden standard of follow up,the efficiency of 3 kinds of assay to discriminate HIV antibody indeterminant were evaluated.Results There were 38 patterns among 394 HIV antibody indeterminant,the proportions of env,pol and gag indeterminant were 37.54%,4.04%and 58.37% respectively.Five HIV antibody indeterminant cases were converted to HIV antibody positive among 97 followed individuals,they were all env indeterminant and HIV antibody developed rapidly.HIV viral load was an ideal assay to discriminate HIV antibody indeterminant with best sensitivity.Conclusion The indeterminant of gag were most common,but were unspecific reaction.Env indeterminant were with the greatest predictive value of HIV infection,especially the gp160p24 and gp160.Viral load assay can be applied to discriminate HIV antibody indeterminant.
2.Effect of Buzhong Yiqiwan on NLRP3 Inflammasome Pathway of DSS-induced Colitis Model Mice at Different Pathological Stages
Chunhui SONG ; Yihui YOU ; Junyu KE ; Geng LI ; Haishan LONG ; Yanli WU ; Qun DU ; Yanwu LI ; Wenfeng GUO
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(14):20-28
ObjectiveTo explore the intervention effect and mechanism of Buzhong Yiqiwan (BZYQ) on colitis mice. MethodSixty-four C57BL/6 mice were randomly divided into 2 weeks blank group, 2 weeks model group, 2 weeks high-dose BZYQ (12 g·kg-1) group, 2 weeks low-dose BZYQ (6 g·kg-1) group, 4 weeks blank group, 4 weeks model group, 4 weeks high-dose BZYQ (12 g·kg-1) group, and 4 weeks low-dose BZYQ (6 g·kg-1) group. The colitis model was induced in mice by feeding 3% dextran sodium sulfate (DSS) for 7 days. The mice received BZYQ (12 and 6 g·kg-1) by gavage on the 8th day after modeling, once per day, and sacrificed on the 2nd and 4th weeks, correspondingly. The colon length and weight of mice in each group were measured. Hematoxylin-eosin (HE) staining was used for pathological observation and colonic mucosal inflammation was scored. The mRNA and protein expression of NOD-like receptor thermoprotein domain 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), and cysteinyl aspartate-specific protease-1 (Caspase-1) was detected by real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot, respectively. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of inflammatory cytokines, such as interleukin (IL)-1β, IL-18, and IL-33 in colonic tissues. ResultCompared with the 2 weeks blank group, the 2 weeks model group showed shortened colon length, increased colon weight (P<0.05), loss of epithelial cells, destruction of gland structure, infiltration of a large number of inflammatory cells in mucosa and submucosa, local crypt abscess, and increase in mucosal inflammation score (P<0.01) as revealed by light microscopy, elevated levels of IL-1β, IL-18, and IL-33 in colonic tissues (P<0.05), and increased mRNA and protein expression of NLRP3, ASC, and Caspase-1 (P<0.05). The intervention of BZYQ (12 g·kg-1) restored colon length, alleviated mucosal injury (P<0.05), down-regulated the content of IL-18 (P<0.05), reduced the mRNA expression of NLRP3 and ASC as well as the protein expression of ASC and Caspase-1 compared with the conditions in the 2 weeks model group. Compared with the 4 weeks blank group, the 4 weeks model group showed decreased colon length, increased colon weight (P<0.05), decreased glands in the mucosal layer, expansion of glandular cavity, atrophy of crypt, local connective tissue hyperplasia and lymphocyte infiltration, increased inflammation score (P<0.01) as revealed by the light microscopy, increased expression of IL-1β, IL-18, and IL-33 (P<0.05), and elevated mRNA and expression of NLRP3 inflammasome complex (P<0.05). Compared with the conditions in the 4 weeks model group, the intervention of BZYQ (12 and 6 g·kg-1) could improve colon length and weight (P<0.05), and the intervention of BZYQ (12 g·kg-1) could also improve the inflammation score of the colon (P<0.05). Different from the acute stage, the intervention of BZYQ (12 and 6 g·kg-1) increased the content of IL-33 in the intestinal mucosa and up-regulated the mRNA and protein expression of NLRP3 inflammasome complexes ASC and Caspase-1 (P<0.05). ConclusionBZYQ can relieve the injury of colitis induced by DSS in mice. The mechanism is related to the regulation of intestinal immune response mediated by NLRP3 inflammasome, and it has different regulatory effects in acute and chronic inflammation stages.