CD4-Positive T-Lymphocytes ; Hepatitis B virus ; Hepatitis B, Chronic ; Humans ; Interleukin-2 Receptor alpha Subunit ; Sequence Analysis

Objective:The recombinant shuttle peptide-single chain antibody with neutralizing activity is expected to be obtained by combining the shuttle peptide sequence with rabies single chain antibody and optimizing the expression conditions in E. coli.Methods:In this study, the SO57 single-stranded antibody sequence was recoded and the shuttle peptide of Arg 12 was fused at the N-terminal to construct pET22 (b)-rSO57 expression vector, which was then expressed in E. coli. The growth density OD 600, induction time, induction temperature and inducer concentration were optimized to obtain a higher expression effect. The recombinant protein was purified by immobilized metal chelating chromatography, and the relative affinity of the recombinant single-chain variable fragment (scFv) was determined. The neutralization effect of the recombinant scFv was verified by rSO57/CVS-11 virus strain mixed with infected cells and rapid fluorescent focus inhibition test (RFFIT). Results:The recombinant rSO57 was successfully constructed and induced in BL21 (de3). The optimal expression condition was that when the cell density of OD 600 was 0.8, IPTG of 0.4 mmol/ml was used for induction. After induction at 16 ℃ for 24 hours, rSO57 was expressed in soluble form, accounting for 19.8% of the soluble protein in the cell. After chromatography purification, the recombinant rSO57 with a purity of 84% was obtained. ELISA showed that the relative affinity coefficient Ka of rSO57 was 4.3×10 5. When rSO57 was mixed with CVS-11 strain, the infection rate of cells increased with the increase of dilution, indicating the neutralization activity of recombinant antibodies. Using the RFFIT, 50 μg rSO57 was equivalent to 2.17 IU of rabies neutralizing serum. Conclusions:In this study, the recombinant scFv fusion with shuttle peptide was expressed, which could neutralize rabies virus, in order to prepare specific and targeted antiviral drug carriers for rabies treatment.

Objective:To find out the chemical constituents in Atrichum undulatum var. gacelisetum.Methods:The compounds were isolated by silica gel and TLC, and purified by recrystallization from the ethanol extract. The structures of the compounds were elucidated by spectral analysis and physicochemical properties. Results:Six compounds were obtained and the structures were identified as dotriacontane ( 1 ), montanic acid ( 2 ), β-sitosterol ( 3 ), daucosterol ( 4 ), luteolin 7-O-D-glucoside ( 5 ) and luteolin ( 6 ). Conclusion:Compounds 1-6 are isolated from the species for the first time.

OBJECTIVETo investigate the role ofCD4+CD25+ T regulatory (Treg) cells, T helper (Th)17cells and interleukin (IL)-6 in the progression of hepatitis B virus (HBV)-related acute-on-chronic liver failure (ACLF) and determine their value as prognostic markers.

METHODSThe Chinese National Knowledge Infrastructure (CNKI), WanFang, Chinese Scientific Journals (VIP), PubMed, Embase and Web of Science databases were searched for English language case-control studies on the relationship between regulatory T lymphocytes and ACLF.The quality of included studies was assessed using the Newcastle-Ottawa scale. The meta-analysis was designed according to the PICOS approach recommended by the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) statement. RevMan software, version 5.1, was used to perform the meta-analysis.

RESULTSNine case-cohort studies were selected for inclusion in the metaanalysis.The results of the meta-analyses showed that the level of CD4+CD25+ Treg cells was not significantly different between patients with HBV-related ACLF and patients with chronic hepatitis B (CHB) (mean difference (MD)=0.59, 95% confidence interval (CI)-1.68, 2.85, P=0.61) nor between patients with HBVrelated ACLF and healthy controls (MD=1.12, 95% CI:-1.42, 3.66, P=0.39). Thus, it appears that ACLF patients do not have a higher level of CD4+CD25+ Treg cells than CHB patients or healthy controls. However, the ACLF patients did appear to have a significantly higher level of Th17 cells than both the CHB patients (MD=1.73, 95% CI:0.21, 3.26, P=0.03) and the healthy controls (MD=1.62, 95% CI:(0.52, 2.72, P=0.004). In addition, the ACLF patients also had significantly higher level than both the CHB patients (MD=11.69, 95%CI:1.98, 21.40, P=0.02) and the healthy controls (MD=13.17, 95% CI:1.38, 24.95, P=0.03).

CONCLUSIONCD4+CD25+ Treg cells may be an important protective factor in the progression and prognosis of HBV-related ACLF, while Thl7 cells and IL-6 may be risk factors for further progression and worsened prognosis.

Acute-On-Chronic Liver Failure ; diagnosis ; immunology ; CD8-Positive T-Lymphocytes ; Case-Control Studies ; Disease Progression ; Hepatitis B virus ; Hepatitis B, Chronic ; complications ; Humans ; Interleukin-6 ; immunology ; Prognosis ; T-Lymphocytes, Regulatory ; immunology ; Th17 Cells ; immunology