Objective: To investigate the neuroprotective effect of hyaluronidase(HAase) on rabbit brain tissue in germinal matrix-intraventricular hemorrhage(GM-IVH) premature rabbits. Methods: Eighty premature rabbits of gestation age 29 days were randomly divided into normal group, GM-IVH control group and HAase treatment group.The rabbits in GM-IVH control group and HAase treatment group were given intraperitoneal injection of 50 g/L glycerol to establish GM-IVH animal model, while the premature rabbits in normal group were given the same dose of 9 g/L saline.The GM-IVH was screened by using cranial ultrasound.The premature rabbits in HAase treatment group were given HAase into the lateral forebrain while the rabbits in the GM-IVH control group were given the same dose of 9 g/L saline.In 3 days, 7 days and 14 days after birth, the premature rabbits were killed and brain tissue were separated.The expression of neuron-glial antigen 2(NG2) were detected by adopting immunohistochemical method.The expression of myelin basic protein(MBP) were detected by Western blot method. Results: The expression of NG2 in 3 groups of premature rabbits decreased gradually with the increase of age[the values of NG2 in the normal group were(62.65±33.58)×10⁴, (15.61±4.22)×10⁴, and (13.54±4.51)×10⁴ on the 3^rd, 7^th, and 14^th day; those in the GM-IVH control group were (54.58±25.48)×10⁴, (48.91±22.49)×10⁴, (7.18±2.28)×10⁴ on the 3^rd, 7^th, and 14^th day; those in the HAase treatment group were (148.13±27.30)×10⁴, (88.38±14.55)×10⁴, (77.98±18.96)×10⁴ on the 3^rd, 7^th, and 14^th day]. At the same time, the expression of NG2 in the HAase treatment group was higher than that of the GM-IVH control group, and the differences were statistically significant among the 3 groups at any time(all P<0.05). The expression of MBP protein in the 3 groups increased with the increase of day age[MBP protein in the normal group were (0.30±0.22)×10³, (1.91±0.43)×10³, and (5.67±2.14) ×10³ on the 3^rd, 7^th, and 14^th day; those in the GM-IVH control group were (0.87±0.12)×10³, (1.15±0.22)×10³ and (2.54±0.69) ×10³ on the 3^rd, 7^th, and 14^th day; those in the HAase treatment group were (0.91±1.01)×10³, (2.25±0.66)×10³, (3.40±1.10)×10³ on the 3^rd, 7^th, and 14^th day]. The difference was statistically significant between the normal group and the HAase treatment group(P<0.05). Conclusions After HAase treatment, the expression of NG2 and MBP can be improved to a certain extent, promoting the development of oligodendrocytes and regeneration of myelin sheath.

Objective To determine the effects of histone deacetylase inhibitor suberoylanilide hydroxamic acid(SAHA) on the cell proliferation and apoptosis of the human hepatic stellate cell line LX-2.The possible underlying mechanisms were also investigated.Methods The LX-2 cells were treated with SAHA in vitro.The morphology of LX-2 cells in different concentrations groups was observed by inverted microscope;the proliferation of LX-2 cells was measured by MTT assay;the Annexin V-FITC and PI staining was used to detect the apoptosis of LX-2 cells by flow cytometry and fluorescence microscope;the expression of α-SMA,collagen Ⅰ,acH3K9,acH3K14 and acH3K18 were detected by Western blot.Results The morphology change of LX-2 cells showed that SAHA inhibited the proliferation rate of LX-2 cells and in a dose dependent manner(P<0.05).The LX-2 cells were sensitive to SAHA along with time increasing,and in a time-dependent manner(P<0.05).Western blot showed that the expression levels of α-SMA and collagen-Ⅰ were significantly lower(P<0.05),on the contrary,the acetylation levels of acH3K9,acH3K14 and acH3K18 were significantly higher (P<0.05).Conclusions The increased acetylation of the histone acH3K9,acH3K14,acH3K18 and the lower expressed α-SMA and collagen-Ⅰ in LX-2 cells may be one of the mechanisms of SAHA.

Objective:To investigate the antiviral efficacy of direct-acting antiviral agents (DAAs) in the treatment of liver transplantation (LT) recipients with hepatitis C virus (HCV) infection.Methods:Twenty-two HCV-infected LT recipients treated with DAAs at Fifth Medical Center of Chinese PLA General Hospital from December 2014 to June 2018 were retrospectively analyzed, Twenty cases of HCV RNA gene type 1b were treated with sofosbuvir (400 mg/d) + ledipasvir (90 mg/d) or sofosbuvir (400 mg/d) + daclatasvir (60 mg/d) for 12 weeks or 24 weeks; 2 cases of gene type 2a were treated with sofosbuvir (400 mg/d) for 12 weeks. The effect of antiviral treatment, adverse reactions during treatment, and laboratory indicators such as HCVRNA quantification, blood routine, liver and kidney function during treatment and follow-up were studied.Results:The LT recipients of HCV infection included 16 males and 6 females, with a median age of 61.5 (36-71) years old, and the median time of antiviral treatment was 48 (2-117) months after transplantation. Among the 22 patients, 16 received a 12-week course of treatment. Except for 2 patients who did not get HCVRNA negative conversion at 4-week, all achieved a negative HCV RNA at 4-week and the end of the treatment. Six LT recipients received a 24-week course of treatment (gene type 1b), and HCVRNA was negative at 4-week and the end of treatment. All patients achieved end of treatment virological response and a sustained virological response (SVR) rate of 100% at 12 weeks and 24 weeks after the end of treatment. The serum levels of alanine aminotransferase (ALT) and creatinine were 71.5 (30, 110) U/L and (89.4±25.7) mmol/L before treatment, respectively. ALT decreased to 22 (17.8, 28.5) U/L after 4 weeks of treatment, and serum creatinine decreased to (77.4±11.5) mmol/L at 24 weeks after the end of treatment. The differences before and after treatment were statistically significant (all P<0.05). No serious adverse events occurred during the treatment. Conclusions:DAAs have a definite antiviral effect in the treatment of LT recipients with HCV infection, and long-term SVR can be obtained.

The present study analyzes the use of the online teaching platform of China Medical University during the COVID-19 epidemic, excavates the teaching difficulties encountered in online teaching, and shares the experience of implementing "1+M" mode of mixed online teaching platform by introducing a variety of online teaching platforms. Monitoring of teaching quality has been initiated on time, and the key tasks of online teaching have been straightened out in time, which has effectively improved the quality of online teaching, providing references and basis for further advancing the reform of higher medical education in China.

Objective To study the effects of efflux pump inhibitors(CCCP and PAβN)on carbapenems in Pseudomonas aernginosa(P.aeruginosa)clinical isolates and investigate the association between the resistance to imipenem or meropenem and expression levels of efflux pumps of P.aeruginosa.Methods MICs of imipenem or meropenem combined with efflux pump inhibitors including carbonyl cyanide m-chlorophenylhydrazone(CCCP,107 strains)and Phe-Arg-β-naphthylamide(PAβN,71 strains)against imipenem-resistant strains were determined by agar dilution method,and changes of MICs were observed.For 32 strains with different resistant phenotypes to imipenem and meropenem,the mRNA expression levels of three efflux pump genes(mexA,mexD and mexF)were quantified by real time fluorescent quantitative PCR.Results The resistance rate of imipenem and meropenem didn't prove any significant difference in the presence of efflux pump inhibitors.The X2 value of imipenem combined with CCCP and PAβN were 0.338 and 0.086,respectively(P＞0.05),while that of meropenem combined with CCCP and PAβN were 1.065 and 1.458(P＞0.05).No significant in MICs of carbapenems were seen in over half of P. aeruginesa isolates. MICs of carbapenems was significantly downregulated for 4-fold or above in eight isolates. Overexpression of efflux pumps genes were present in 24 of 27 carbapenem-resistant isolates(88. 9% ). Efflux pumps genes including MexAB-OprM, MexCD-OprJ and MexEF-OprN were all overexpressed in 13 isolates,constituting 54. 2% of all carbapenem-resistant isolates. There were 3 isolates in which beth MexAB-OprM and MexCD-OprJ showed overexpression,constituting 12. 5%. Also,MexAB-OprM and MexEF-OprN overexpressed in 3 isolates. There were 2 isolates (8.3%) showing MexEF-OprN overexpression and MexAB-OprM alone. MexCD-OprJ didn't showed overexpression alone. Furthermore,the expression levels of efflux pumps genes mexA,mexD and mexF in isolates susceptible to both in imipenem and meropenem were 0. 48±0. 48,0. 48±0. 53 and 0. 30±0. 41,respectively,which were much lower than that in carbapenem-resistant ones (P＜0. 05 ). MexA gene was expressed at a higher level in meropenemresistant isolates than meropenem-susceptible ones (P＜0. 05 ). Conclusions When the concentration of CCCP and PAβN were 5 μg/ml and 20 μg/ml respectively,the efforts on the carhapenems resistance of P.aeruginosa were small Overexpression of MexAB-OprM might play an important role in meropenemresistance in P. aerugines. Overexpression of MexCD-OprJ and MexEF-OprN was associated with imipenemresistance. However,the relationship between them and meropenem-resistance need to be explored in the future.

BACKGROUND:Prolonged therapy with lamivudine has been associated with tyrosine-methionine-aspartate-aspartate mutation, which results in hepatitis B recurrence. Recently, antiviral agents, such as entecavir, have high efficacy and low resistance rate in hepatitis B-related liver disease. However, the researches on the effect of entecavir in preventing hepatitis B recurrence after liver transplantation are rare. OBJECTIVE:To investigate the effect of entecavir combined with low-dose hepatitis B immunoglobulin in preventing hepatitis B recurrence after liver transplantation. METHODS:The fol ow-up data of 253 patients who had liver transplantation for hepatitis B virus related liver disease were retrospectively analyzed. Al patients received nucleoside analogues therapy formal y before liver transplantation. The effects of entecavir+hepatitis B immunoglobulin and lamivudine+hepatitis B immunoglobulin were compared in al the patients and the patents with hepatitis B recurrence risk factors (positive preoperative HBeAg, DNA-positive hepatitis B virus, hepatoma and tyrosine-methionine-aspartate-aspartate mutation). RESULTS AND CONCLUSION:A total of 253 patients received hepatitis B virus-related liver transplantation, and 29 patients died. There were 202 patients in lamivudine group in which 26 patients were dead and 16 patients had hepatitis B virus recurrence, and the recurrence rate was 7.92%(16/202). However, entecavir group had 51 patients without hepatitis B virus recurrence in which three patients were dead. There were significant differences in the mortality rate and recurrence rate between two groups. Compared with the lamivudine+hepatitis B immunoglobulin, entecavir+hepatitis B immunoglobulin could effectively reduce the recurrence rate of the patients with hepatitis B virus-related risk factors. Hepatitis B immunoglobulin was terminated and nucleoside analogues were modulated when recurrence appeared. Al patients hepatitis B virus DNA were control ed less than 500 IU/mL and liver function returned to normal level. Log-rank test showed that there was no significant difference in the long-term survival rate after timely treatment of hepatitis B virus recurrence. With the prevention of nucleoside analogues combined with hepatitis B immunoglobulin therapy, timely treatment of hepatitis B recurrence has little influence on the prognosis. Entecavir combined with hepatitis B immunoglobulin can effectively prevent the hepatitis B recurrence. For the patients with hepatitis B virus-related risk factors, entecavir combined with hepatitis B immunoglobulin can better reduce the recurrence rate of hepatitis B than lamivudine+hepatitis B immunoglobulin after liver transplantation.

AIM:To study the effects of suberoylanilide hydroxamic acid (SAHA) on the apoptosis of hepatic stellate cells (HSCs) and expression of associated proteins, and to investigate the mechanisms of SAHA to induce apoptosis.METHODS:The rat HSCs were isolated by OptiPrep gradient centrifugation method.The effect of SAHA on HSC proliferation was detected by real-time cell analyzer.The morphological changes of HSCs treated with SAHA at different concentrations were observed under inverted microscope.The apoptotic rates of HSCs were analyzed by flow cytometry with Annexin V-FITC/PI staining and fluorescence microscopy.The protein expression of α-smooth muscle actin (α-SMA), collagen I, tissue inhibitor of metalloproteinase 1 (TIMP1), glucose-regulated protein 78 (GRP78) and histone deacetylase 6 (HDAC6) was detected by Western blotting.The interaction of GRP78 with HDAC6 in the HSCs was determined by co-immunoprecipitation.RESULTS:HSCs were successfully isolated and cultured for 14 d, during which the HSCs changed gradually from rest state to active state.SAHA significantly inhibited the proliferation of HSCs in a time-and dose-dependent manner (P<0.05).The results of Western blotting showed that the protein expression levels of α-SMA, TIMP1, collagen-I and HDAC6 were significantly decreased (P<0.05), while GRP78 was significantly increased (P<0.05).Compared with activated HSCs, GRP78 and total acetyl-lysine protein were significantly increased in the co-immunoprecipitated HSCs treated with SAHA, while HDAC6 protein was significantly decreased, indicting that GRP78 formed a complex with HDAC6.CONCLUSION:The anti-hepatic fibrosis effect of SAHA may be related to down-regulation of HDAC6 and up-regulation of acetylated GRP78, thus inducing endoplasmic reticulum stress of HSCs and promoting the apoptosis of HSCs.

With the approval and launch of a large number of new drugs, the incidence rate of drug-induced liver injury (DILI) is increasing year by year, which may affect the treatment of primary diseases. As an adverse drug reaction, DILI cannot be completely eliminated, and the clinical goal is to minimize its influence through prevention and control. This article reviews the research advances in the risk factors for DILI, the monitoring of DILI, and retreatment. Studies have shown that the risk of DILI can be increased by certain factors under some circumstances. Early identification of risk factors, rational monitoring, and focus on the timing and method for retreatment can reduce the development or progression of DILI and thus improve the prognosis of patients.

Objective:To investigate the current situation and analyze the influencing factors of job satisfaction of infectious disease physicians in Jiangsu Province.Methods:From April to May 2021, a stratified random sampling method was used to select 10 municipal level infectious disease hospitals in Jiangsu province. An electronic questionnaire survey was conducted on the infectious disease physicians of these hospitals using an online questionnaire platform. The questionnaire mainly included the job satisfaction scale developed by Greenhaus and Wormley, and a self-designed job satisfaction influencing factor scale based on resource conservation theory. At the same time, semi-structured interviews were made with relevant insiders on such issues as salary and benefits, and career development among others. Descriptive analysis was conducted on the questionnaire data, while single factor analysis and multiple linear regression analysis were used to identify the factors that affect their job satisfaction, and the results of the interviews were studied with Colaizzi 7-step analysis.Results:A total of 457 valid questionnaires were recovered. The job satisfaction score of infectious disease physicians was (3.38±0.76). The results of univariate analysis showed significant differences in job satisfaction scores among infectious disease physicians of different ages, educational backgrounds, professional titles, and positions. The results of multiple linear regression analysis showed that salary and benefits ( B=0.141, P<0.001), work environment ( B=0.128, P<0.001), leadership support ( B=0.112, P=0.014), development security ( B=0.117, P<0.001), self-efficacy ( B=0.084, P=0.045) and social support ( B=0.285, P<0.001) were both influencing factors of their job satisfaction. The interview results identified such problems as low salary, insufficient salary security, large gap in social security levels between in-staff and off-staff physicians, and insufficient career development potential. Conclusions:The job satisfaction of infectious disease physicians in Jiangsu Province was found at an average level. It is recommended that the government increase the investment in personnel expenses for infectious disease physicians, optimize the salary and benefits structure, pay attention to the career development needs of middle-aged and young physicians, and care about the social relationship needs of infectious disease physicians to improve their job satisfaction.

ObjectiveTo investigate the risk factors for tumor recurrence and death after liver transplantation in patients with hepatocellular carcinoma (HCC) and their survival. MethodsThe patients with HCC who underwent liver transplantation in The Fifth Medical Center of Chinese PLA General Hospital from January 2005 to February 2019 were enrolled, and according to the presence or absence of HCC recurrence after liver transplantation, they were divided into recurrence group and non-recurrence group. The t-test or the Mann-Whitney U test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. Univariate and multivariate Cox proportional-hazards regression model analyses were used to determine the risk factors for HCC recurrence and death after liver transplantation. The Kaplan-Meier method was used for survival analysis, and the receiver operating characteristic (ROC) curve was used to investigate the predictive value of death-related risk factors after liver transplantation. ResultsA total of 391 HCC patients who underwent liver transplantation were enrolled, with a median follow-up time of 2 years, among whom 78(19.95%) experienced HCC recurrence. Preoperative alpha-fetoprotein (AFP) level＞200 ng/ml (recurrence: hazard ratio ［HR］=252, 95% confidence interval ［CI］: 1.58-4.03, P＜0.001; death: HR=2.99, 95%CI: 1.59-5.62, P＜0.001］, total tumor diameter (recurrence: HR=1.20, 95%CI: 1.12-1.28, P＜0.001; death: HR=1.10, 95%CI: 1.02-1.17, P=0.002), and vascular invasion (recurrence: HR=1.15, 95%CI: 1.04-1.26, P=0.016; death: HR=1.10, 95%CI: 1.03-1.18, P=0.004) were independent risk factors for tumor recurrence and death after liver transplantation. The 1-, 5-, and 10-year overall survival rates after liver transplantation were 94.8%, 84.2%, and 83.5%, respectively, and the 1-, 5-, and 10-year disease-free survival rates were 840%, 75.1%, and 75.1%, respectively. AFP, involvement of major blood vessels, body mass index, and total tumor diameter had a certain value in predicting the death of HCC patients with recurrence, with an area under the ROC curve of 0.789 (95% CI: 0.719-0858). ConclusionTumor biological features before transplantation are the key factors for tumor recurrence after transplantation.