HIV-1 integrase (IN) is a key enzyme for the viral replication. The protein-protein interaction (PPI) between HIV-1 IN and a cellular cofactor lens epithelium-derived growth factor (LEDGF/p75) is a validated target for anti-HIV drug discovery. In order to build the platform for screening inhibitor against PPI between IN and LEDGF/p75, the vector containing the LEDGF/p75 protein cDNA was constructed and expressed in Escherichia coli and the function of the LEDGF/p75 protein was assayed. The LGDGF/p75 encoding gene optimized according to the preference codon usage of E. coli, was synthesized and cloned into the expression vector pGEX-4T-1 to form a recombined plasmid, then transformed into host cell E. coli BL21 (DE3). The recombined clones were identified and confirmed by BamH I/Sal I digestion and sequencing, the successfully recombined plasmid in the host cell was induced by IPTG and the condition of the expression was optimized. The expressed protein was purified by the Ni2+ affinity chromatography column and SDS-PAGE was used to analyze the molecular weight and specificity. In addition, ELISA assay was used to analyze the function of the recombinant protein. The recombinant LGDGF/p75 was soluble, and expressed highly and stably in E. coli. The protein was proved to enhance HIV-1 IN strand transfer activity in vitro by ELISA. It will be helpful to build the platform of screening inhibitors against PPI between IN and LEDGF/p75.

Objective To determine the expression of human protection of telomeres 1 (hPOT1) and its relationship with Helicobacter pylori (HP) infection in gastric carcinoma.Methods The expressions of hPOT1 protein and hPOT mRNA were detected in 53 gastric carcinoma specimens (observed group) and 20 normal gastric mucosa tissues (control group) by SP immunohistochemical method and in situ hybridization (ISH), respectively.HP infection was examined by Warthin-Starry method in observed group.Results The positive expression rate of hPOT1 protein was 84.91％ (45/53) in observed group, higher than that in control group [30.00 ％ (6/20)] (P ＜ 0.01).The positive expression rate of hPOT1 mRNA was 58.49 ％ (31/53) in observed group, higher than that in control group [10.00 ％ (2/20)] (P ＜ 0.05).The positive co-expression rate of hPOT1 protein and mRNA was 56.60 ％ (30/53), both had positive relationship in gastric carcinoma (r =0.394, P ＜ 0.05).The rate of HP infection in 53 cases of gastric carcinoma was 52.83 ％ (28/53).The positive expression rates of hPOTI protein and mRNA in observed group with HP infection were significantly higher than those in observed group without HP infection[protein: 96.43 ％ (27/28) vs 72.00 ％ (18/25), P ＜0.05;mRNA: 85.75 ％ (24/28) vs 28.00 ％ (7/25), P ＜ 0.01].Conclusions hPOT1 may be associated with occurrence of gastric carcinoma.Combined detection of hPOT1 protein and mRNA can be used for the diagnosis of gastric carcinoma.HP infection may be associated with abnormal expression of hPOT1 in occurrence of gastric carcinoma.

Objective To explore the effect of early phase rehabilitation training on the urination function recovery of the patients with paraplegia caused by spinal cord injury (SCI).Methods Sixty-six patients with paraplegia caused by SCI were selected and divided into the rehabilitation and the control group.The rehabilitation group of patients received early phase rehabilitation training on the urination function,the control group received routine training on the urination training and urinary catheter nursing care.The urination function recovery effect was compared between two groups.Results The urination function recovery effect of the rehabilitation group was significantly better than the control group.Conclusions The usage of early phases of urination training measures on the SCI paraplegia patients can help them cast off the catheter,build up regular urination,and reduce complications.

The drug resistant mutations in human immunodefieiency virus type 1 (HIV-1) are a major impediment to successful highly active antiretrovirai therapy (HAART) and new drug design. In order to understand the drug resistance mechanism of HIV-1 integrase (IN) mutually existed for multiple drug-resistant strains to the most potent IN inhibitors diketo acids (DKAs), three S-1360-resistant HIV-1 strains were selected and molecular docking and molecular dynamics (MD) simulations were performed to obtain the inhibitor binding modes. Based on the binding modes, compelling differences between the wild-type and the 3 mutants for IN have been observed. The results showed that: 1) In the mutants, the inhibitor is close to the funetional loop 3 region but far away from the DNA binding site. Different binding sites lead to the decrease in susceptibility to S-1360 in mutants compared to the wild-type IN. 2) The fluctuations in the region of residues 138～166 are important to the biological function of IN. 2 hydrogen-bonds between S-1360 with residues N155 and K159 restrict the flexibility of the region. Drug resistant mutations result in a lack of the interaction, consequently, the less susceptible to S-1360. 3) In the 3 mutant IN complexes, the benzyl ring of S-1360 is far from the viral DNA binding site, thus, S-1360 can not prevent the end of the viral DNA from exposure to human DNA. 4) After T66I mutation, the long side chain of I occupied the active pocket in the 3 mutants, consequently, the inhibitor could not move into the same binding site or have the same orientation. All the above contribute to drug resistance. These results will be useful for the rational inhibitor modify and design.

Lung ; pathology ; Pneumoconiosis ; microbiology ; Pseudomonas Infections ; microbiology ; Pseudomonas aeruginosa

Objective: To analyze the clinical characteristics of cases of novel coronavirus pneumonia and a preliminary study to explore the relationship between different clinical classification and liver damage. Methods: Consecutively confirmed novel coronavirus infection cases admitted to seven designated hospitals during January 23, 2020 to February 8, 2020 were included. Clinical classification (mild, moderate, severe, and critical) was carried out according to the diagnosis and treatment program of novel coronavirus pneumonia (Trial Fifth Edition) issued by the National Health Commission. The research data were analyzed using SPSS19.0 statistical software. Quantitative data were expressed as median (interquartile range), and qualitative data were expressed as frequency and rate. Results: 32 confirmed cases that met the inclusion criteria were included. 28 cases were of mild or moderate type (87.50%), and four cases (12.50%) of severe or critical type. Four cases (12.5%) were combined with one underlying disease (bronchial asthma, coronary heart disease, malignant tumor, chronic kidney disease), and one case (3.13%) was simultaneously combined with high blood pressure and malignant tumor. The results of laboratory examination showed that the alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (ALB), and total bilirubin (TBil) for entire cohort were 26.98 (16.88 ~ 46.09) U/L and 24.75 (18.71 ~ 31.79) U/L, 39.00 (36.20 ~ 44.20) g/L and 16.40 (11.34- ~ 21.15) mmol/L, respectively. ALT, AST, ALB and TBil of the mild or moderate subgroups were 22.75 (16.31- ~ 37.25) U/L, 23.63 (18.71 ~ 26.50) U/L, 39.70 (36.50 ~ 46.10) g/L, and 15.95 (11.34 ~ 20.83) mmol/L, respectively. ALT, AST, ALB and TBil of the severe or critical subgroups were 60.25 (40.88 ~ 68.90) U/L, 37.00 (20.88 ~ 64.45) U/L, 35.75 (28.68 ~ 42.00) g/L, and 20.50 (11.28 ~ 25.00) mmol/L, respectively. Conclusion The results of this multicenter retrospective study suggests that novel coronavirus pneumonia combined with liver damage is more likely to be caused by adverse drug reactions and systemic inflammation in severe patients receiving medical treatment. Therefore, liver function monitoring and evaluation should be strengthened during the treatment of such patients.