This study explored the effect of the Mycobacterium tuberculosis(Mtb)PE/PPE family protein PE_PGRS37 on the growth of Mycobacterium smegmatis(Ms)and macrophage autophagy during Mtb infection.The pe_pgrs37 gene was amplified from Mtb genome through PCR,and the recombinant vector pAIN-PE_PGRS37 was successfully constructed through homologous recombi-nation.pAIN-PE_PGRS37 and pAIN were integrated into Ms through electroshock to construct pAIN-PGRS37/Ms and pAIN/Ms re-combinant bacteria.Western blotting indicated that the PE_PGRS37 protein was correctly expressed in pAIN-PE_PGRS37/Ms.The re-combinant bacteria were inoculated in 7H9/7H10 medium,and their colony morphology and growth curves were observed.No signifi-cant difference in colony morphology was observed between pAIN-PE_PGRS37/Ms and pAIN/Ms.The growth rate significantly in-creased between 10 and 16 h,and a plateau was reached at 26 h.After infection of U937 cells with pAIN-PE_PGRS37/Ms and pAIN/Ms,macrophage autophagy flow was detected with western blotting and immunofluorescence.In the pAIN-PE_PGRS37/Ms-infected group,compared with the pAIN/Ms-infected group,macrophage LC3-II and p62 protein expression was significantly up-regulated(P<0.001)and inhibited autophagosome and lysosome fusion.The intracellular survival of the recombinant bacteria was detected through colony counting,and pAIN-PE_PGRS37/Ms showed significantly greater survival in macrophages at 12 h,24 h,and 48 h than pAIN/Ms(P<0.05).Our results suggested that PE_PGRS37 protein promotes Mycobacterium survival in macrophages by blocking macro-phage autophagy flow,thus inhibiting macrophage autophagy.

This study explored the effect of the Mycobacterium tuberculosis(Mtb)PE/PPE family protein PE_PGRS37 on the growth of Mycobacterium smegmatis(Ms)and macrophage autophagy during Mtb infection.The pe_pgrs37 gene was amplified from Mtb genome through PCR,and the recombinant vector pAIN-PE_PGRS37 was successfully constructed through homologous recombi-nation.pAIN-PE_PGRS37 and pAIN were integrated into Ms through electroshock to construct pAIN-PGRS37/Ms and pAIN/Ms re-combinant bacteria.Western blotting indicated that the PE_PGRS37 protein was correctly expressed in pAIN-PE_PGRS37/Ms.The re-combinant bacteria were inoculated in 7H9/7H10 medium,and their colony morphology and growth curves were observed.No signifi-cant difference in colony morphology was observed between pAIN-PE_PGRS37/Ms and pAIN/Ms.The growth rate significantly in-creased between 10 and 16 h,and a plateau was reached at 26 h.After infection of U937 cells with pAIN-PE_PGRS37/Ms and pAIN/Ms,macrophage autophagy flow was detected with western blotting and immunofluorescence.In the pAIN-PE_PGRS37/Ms-infected group,compared with the pAIN/Ms-infected group,macrophage LC3-II and p62 protein expression was significantly up-regulated(P<0.001)and inhibited autophagosome and lysosome fusion.The intracellular survival of the recombinant bacteria was detected through colony counting,and pAIN-PE_PGRS37/Ms showed significantly greater survival in macrophages at 12 h,24 h,and 48 h than pAIN/Ms(P<0.05).Our results suggested that PE_PGRS37 protein promotes Mycobacterium survival in macrophages by blocking macro-phage autophagy flow,thus inhibiting macrophage autophagy.

Objective To study the expression and significance of Glypican-3 in Budd-Chiari syndrome (BCS) complicated with hepatocellular carcinoma (HCC).Methods The data of 46 patients with BCS complicated with HCC (the BCS + HCC group) treated in The First Affiliated Hospital of Zhengzhou University from January 2007 to December 2016 were analyzed retrospectively.Another 48 patients with HBV-related HCC (the HBV + HCC group) and 43 patients with hepatic cyst (the hepatic cyst group) were randomly selected as the control groups during the same time period.The differencesin positive rates of Glypican-3 in the liver tissues among the three groups were compared.The BCS + HCC group was further divided into the Glypican-3 positive and Glypican-3 negative subgroups according to the expression of Glypican-3.The differences in gender,age,AFP,HbsAg,Child-Pugh classification,tumor number,extrahepatic metastasis,vascular invasion,Edmondson-Steiner grading and BCLC staging between the two subgroups were compared.The survival time of the two subgroups was compared using the Kaplan-Meier method.Results The expression rates of Glypican-3 in the BCS + HCC group,HBV + HCC group and Hepatic Cyst group were 76.1％,70.8％ and 0％,respectively.The levels of Glypican-3 in the BCS + HCC group and the HCC group were significantly higher than that in the hepatic cyst group.The differences were statistically significant (P ＜ 0.05).No statistically significant difference was detected between the BCS + HCC group and the HBV + HCC group (P ＞ 0.05).In the group of patients with BCS + HCC,there was no significant difference in gender,age,AFP,HbsAg,Child-Pugh classification,tumor number and extrahepatic metastasis between the Glypican-3 positive and negative subgroups (P ＞0.05).However,vascular invasion,Edmondson-Steiner grading and BCLC staging in the Glypican-3 positive subgroup were significantly higher than those in the Glypican-3 negative group,(P ＜ 0.05).The 1-year,3-year and 5-year survival rates were 77.1％,51.0％ and 22.8％ in the Glypican-3 positive subgroup,compared with 90.9％,63.6％ and 45.5％ in the Glypican-3 negative subgroup,respectively.There were statistically significant differences between the two groups (P ＜ 0.05).Conclusion Glypican-3 has a stable expression in patients with BCS complicated with HCC,and it is closely related to malignancy of the tumor and prognosis of the patients.

Objective:To quantitively assess the mental status of patients with severe acute respiratory syndrome(SARS) for correct diagnosis of those have serious mental crisis,providing scientific basis for the psychological intervention. Methods:The mental status in 46 SARS patients were analyzed with the Mental Status Scale in Non psychiatric Settings(MSSNS, by Department of Psychology, Second Military Medical University )and compared with that of the common pneumonia patients and the norm of common inpatients. Results:The scores of SARS patients and those of the other pneumonia patients and the norm of common inpatients had no significant difference. The psychological reactions of SARS patients were related significantly with the patient's age, marital status, occupation and the duration of fever. Conclusion:The psychological reactions of SARS patients were accordant with those of the common patients. The results indicated that the patient's personality, such as the emotional stability, was the essential impact factor on the patient's mental status. The level of negative emotional reactions in SARS patients are influenced by whether the patients are young, single, lack of medical knowledge and fever duration. So the effective psychological intervention to the SARS patients should be based on the scientific assessment and influencing factors analysis.