Objective:To investigate the antimicrobial activity of the antiparasitic drug tiliquinol to Enterococcus faecalis. Methods:From 2023 to 2024, the standard Enterococcus faecalis strain ATCC 29212 and 6 clinical isolates in the Laboratory Department of the Affiliated Changsha Hospital of Xiangya School of Medicine were selected as the subject of this investigation. The sensibility of tiliquinol against E. faecalis was assessed using broth microdilution method, time-killing curves, and kirby-bauer test; the drug resistance induction ability of tiliquinol was detected by continuous induction of resistance and one-step resistance test. The antimicrobial ability of tiliquinol was determined by the biofilm combined laser confocal microscopy. Skin subcutaneous abscess model of E. faecalis infection was established to evaluate the in vivo antibacterial activity of tiliquinol. Paired t-tests and analysis of variance were used for comparisons between two groups and among multiple groups respectively. Results:The minimum inhibitory concentration (MIC) and minimum bactericidal concentration of tiliquinol against E. faecalis ATCC 29212 were both 2 mg/L. Kirby-bauer test showed obvious antimicrobial activity of tiliquinol against E. faecalis. The time-killing curves showed that the subinhibitory concentration of tiliquinol could effectively inhibit bacterial proliferation. Fifteen-day continuous treatment with tiliquinol showed no drug resistance mutation; tiliquinol treatment at 8×MIC for four hours caused reduced count of viable bacteria from (12.01±1.14) lg CFU/ml to (7.72±0.94) lg CFU/ml in ATCC 29212 stain ( t=3.64; P<0.05), while tiliquinol at 2×MIC significantly inhibited the formation of ATCC 29212 biofilm, which reduced from (1.73±0.27) to (0.18±0.14) ( t=8.77, P<0.05); tiliquinol at 4×MIC also significantly cleared the formed biofilm, which reduced from (0.52±0.03) to (0.40±0.06) ( t=3.07, P<0.05). Utilizing the skin subcutaneous abscess model revealed significant antibacterial effects of tiliquinol treatment, specifically, and compared with the control group, the viable bacterial loads in the 30 mg/kg tiliquinol treatment group decreased by more than 1 lg CFU/ml ( t=4.099, P<0.05). Conclusion:Tiliquinol exhibits substantial antibacterial efficacy against E. faecalis both in vitro and in vivo.

Objective:To investigate the antimicrobial activity of the antiparasitic drug tiliquinol to Enterococcus faecalis. Methods:From 2023 to 2024, the standard Enterococcus faecalis strain ATCC 29212 and 6 clinical isolates in the Laboratory Department of the Affiliated Changsha Hospital of Xiangya School of Medicine were selected as the subject of this investigation. The sensibility of tiliquinol against E. faecalis was assessed using broth microdilution method, time-killing curves, and kirby-bauer test; the drug resistance induction ability of tiliquinol was detected by continuous induction of resistance and one-step resistance test. The antimicrobial ability of tiliquinol was determined by the biofilm combined laser confocal microscopy. Skin subcutaneous abscess model of E. faecalis infection was established to evaluate the in vivo antibacterial activity of tiliquinol. Paired t-tests and analysis of variance were used for comparisons between two groups and among multiple groups respectively. Results:The minimum inhibitory concentration (MIC) and minimum bactericidal concentration of tiliquinol against E. faecalis ATCC 29212 were both 2 mg/L. Kirby-bauer test showed obvious antimicrobial activity of tiliquinol against E. faecalis. The time-killing curves showed that the subinhibitory concentration of tiliquinol could effectively inhibit bacterial proliferation. Fifteen-day continuous treatment with tiliquinol showed no drug resistance mutation; tiliquinol treatment at 8×MIC for four hours caused reduced count of viable bacteria from (12.01±1.14) lg CFU/ml to (7.72±0.94) lg CFU/ml in ATCC 29212 stain ( t=3.64; P<0.05), while tiliquinol at 2×MIC significantly inhibited the formation of ATCC 29212 biofilm, which reduced from (1.73±0.27) to (0.18±0.14) ( t=8.77, P<0.05); tiliquinol at 4×MIC also significantly cleared the formed biofilm, which reduced from (0.52±0.03) to (0.40±0.06) ( t=3.07, P<0.05). Utilizing the skin subcutaneous abscess model revealed significant antibacterial effects of tiliquinol treatment, specifically, and compared with the control group, the viable bacterial loads in the 30 mg/kg tiliquinol treatment group decreased by more than 1 lg CFU/ml ( t=4.099, P<0.05). Conclusion:Tiliquinol exhibits substantial antibacterial efficacy against E. faecalis both in vitro and in vivo.

Objective: To investigate the clinical efficacy of daytime continuous blood purification (DCRRT) combined with plasma exchange in the treatment of severe acute pancreatitis. Methods: The clinical data of 49 patients with non-biliary severe acute pancreatitis admitted to the First People's Foshan Hospital from January 2012 to January 2019 were analysed respectively. The enrollees were randomized into DCRRT combined with plasma exchange (combination therapy) group and DCRR only (DCRR) group using a random number table method. All patients received DCRRT therapy [8 hours continuous venous-venous blood purification/day (CVVH/d)] immediately after the diagnosis of non-biliary severe acute pancreatitis was established. The combination group received at least one plasma exchange during the course of treatment. The differences of laboratory examination and prognosis between the two groups before and after treatment were compared. Results: A total of49 patients were enrolled, including 29 males and 20 females, with age of (46.40±17.81) years. There were 24 patients in the combination therapy group and 25 patients in DCRR group. There were no significant differences in the age, gender, body mass index (BMI), and pre-treatment laboratory findings between the two groups. After treatment, the blood glucose, hypersensitive C-reactive protein (hs-CRP), procalcitonin (PCT-u), amylase, lipase, triglyceride, cholesterol, serum creatinine were lower than those before treatment (all P<0.05). The blood hs-CPR, PCT-u, lipase and triglyceride in the combination therapy group were significantly lower than those in the DCRR group (all P<0.05). The acute physiology and chronic health scores (APACHEⅡ) of the two groups were lower than those before treatment, and the combination therapy group was more significant than DCRR group (all P<0.05). There were 5 deaths (20.83%) in the combination therapy group and 7 deaths (28.00%) in DCRR group. There was no significant difference in mortality between the two groups. There was no significant difference in the start time and duration of DCRRT between the two groups. Conclusions DCRRT or combined plasma exchange therapy can effectively treat non-biliary severe acute pancreatitis. DCRRT combines with plasma exchange therapy can more effectively remove inflammatory factors and reduce APACHEⅡ score.

Objective To assess the medium and long term efficacy of hemodialysis combined with hemo-perfusion on the endothelial function in patients with maintance hemodialysis(MHD). Methods 60 stable MHD patients were enrolled in the research and randomly divided into 2 group. The observation group received hemodialy-is combined blood perfusion,and the control group received pure hemodialysis therapy. Blood was collected before and after treatment for 6 months for detection of serum C-reactive protein (CRP),hemoglobin (HB),albumin (ALB),advanced glycation end products(AGEs),homocysteine(Hcy)and intercellular cell adhesion molecule (ICAM). Results Plasma hs-CRP,AGEs,Hcy and ICAM decreased gradually after the treatment for 6 months. Compared with the indexes before treatment ,serum HGB and ALB increased significantly after the treatment for 6 months(P < 0.05). Conclusions Hemodialysis combined with hemoperfusion with an appropriate frequency and in a medium or long period is a safe ,convenient,and effective approach for MHD patients to pretect the endotheli-al function.

Objective To investigate mutations in immediate early ( IE) gene ORF62 of three var-icella vaccine Oka strains ( vOka ) including two strains produced in China and their parental Oka strain (pOka), and then to further elucidate its possible roles in attenuation mechanism by comparing their ORF 62 promoter sequences and its activities , ORF62 coding regions and its transactivities .Methods ORF62 pro-moter-reporter plasmids and ORF62-expressing plasmids of pOka and three vOka strains ( vOka-BK from Changchun BCHT Biotechnology Co ., vOka-SH from Shanghai Institute of Biological Product Co .Ltd., and vOka-GSK from GlaxoSmithKline plc, as control) were constructed, respectively.ORF62 promoter regions and coding regions of the four strains were sequenced and then compared with each other .Differences of ac-tivities of the ORF62 promoter, and transactivities of the ORF62-encoded IE62 upon immediate early (ORF4), early (ORF28) and late (ORF67) gene promoters between pOka and vOka strains were assayed with transient transfection technique .Results Compared with pOka strain , three vOka strains had a con-sistent T deletion mutation at site 110 050 in ORF62 promoters, which did not result in any change of tran-scription factor binding motif .However , activities of ORF62 promoters from three vOka strains were signifi-cantly lower than those of pOka strain .Three consistent substitution mutations were observed in ORF 62 cod-ing regions of three vOka strains and three new enzyme restriction sites including SmaⅠ, NaeⅠand BssHⅡwere generated, respectively.Transactivities of IE62 from three vOka strains upon ORF4, ORF28 and ORF67 promoters were significantly higher than those of pOka both in CV-1 and MeWo cells , except that vOka-SH IE62 showed significantly lower transactivities upon ORF 4 promoter than those of pOka strain in CV-1 cells.Conclusion Consistent T deletion mutation at site 110 050 in ORF62 promoters of three vOka strains might be responsible for the reduced promoter activities and the changes of IE 62 transactivities .How-ever , it seemed that cell types have no significant effect on ORF 62 promoter activity or IE 62 transactivity be-tween pOka and vOka strains .

Objective To establish a novel method to screen for anti-varicella-zoster virus (VZV) compounds with our previously generated reporter cell line for VZV, MV9G. MethodsMV9G cells were directly infected with cell-free virus of Oka vaccine strain (vOka) for 2 hours( CFV direct-infection) or cocultured with vOka-infected MeWo cells containing cell-associated virus for 48 hours (CAV co-culture) to promote expression of the reporter gene firefly luciferase. Antiviral compounds including heparin, mannose-6-phosphate( M-6-P), acyclovir( ACV ), resveratrol and roscovitine were added in the medium before or after the virus infection. Inhibitory effects( IC50 ) of the antiviral compounds were analyzed by comparing firefly luciferase activities of MV9G cells in the presence of antiviral compounds with those in the absence. Results Antiviral compounds inhibited luciferase activities of MV9G cells activated by CFV direct-infection and/or CAV co-culture in different levels. The reductions of luciferase activities statistically correlated with those of viral foci shown by immunostaining with a monoclonal antibody against VZV immediate early 62 antigens (IE62) in controls. Among these compounds, heparin, M-6-P, and 2.5 μmol/L of roscovitine inhibited CFV-activated more strongly than CAV-activated luciferase activities, whereas ACV and resveratrol inhibited CAV-activated more strongly than CFV-activated luciferase activities. Cell-associated ACV-resistant strains,Kanno and rOka YSR, activated luciferase activities of MV9G cells, too. However, the inhibitory concentrations (IC50) of ACV to the ACV-resistant strains were much higher than those to the ACV-sensitive strains,pOka and CaGu. ConclusionThe CFV direct-infection and CAV co-culture assays were useful to screen for antiviral compounds targeting the early and late phases of VZV infection, respectively. The VZV reporter cell-based assays may provide a simple, rapid, sensitive, and high-throughput method to screen for anti-VZV compounds.