Objective:To explore the correlation between triglyceride-glucose (TyG) index-body mass index (BMI) product (TyG-BMI) and hypertension.Methods:Based on the cross-sectional survey data of adult hypertension prevalence in Hunan Province from June 2013 to May 2014, 4 012 subjects aged ≥18 years with complete key data were included. Binary logistic regression analysis was used to analyze the correlation between TyG-BMI and hypertension, and receiver operating characteristic (ROC) curve was used to compare the ability of TyG index and TyG-BMI to identify hypertension.Results:A total of 4 012 subjects were included in this study, with an average age of (54.6±12.6)years, and males accounted for 40.98%(1 644/4 012). The prevalence of hypertension was 38.33%(1 538/4 012). Logistic regression analysis showed that elevated TyG-BMI was an independent risk factor for hypertension ( P<0.05). In the fully adjusted model, each 1/4 increase in TyG-BMI was associated with a 1.017-fold increase in the risk of hypertension ( OR=1.017, 95% CI: 1.014-1.019). Compared with the lowest quartile group (Q 1), the higher quartile groups (Q 2, Q 3, Q 4) of TyG-BMI had a higher risk of hypertension, with OR values of 1.841, 2.265, and 4.386, respectively. Restricted cubic spline plot showed a linear dose-response relationship between TyG-BMI and the risk of hypertension (overall trend P<0.001). In subgroup analyses stratified by age, gender, smoking and drinking status, TyG-BMI was positively correlated with hypertension. In addition, ROC curve analysis showed that TyG-BMI had better diagnostic value for hypertension compared with TyG index. Conclusions:TyG-BMI is an independent risk factor for hypertension. Excessively high TyG-BMI or gradual increase of TyG-BMI will increase the risk of hypertension, and TyG-BMI has higher value in identifying hypertension compared with TyG index.

Objective:To explore the correlation between triglyceride-glucose (TyG) index-body mass index (BMI) product (TyG-BMI) and hypertension.Methods:Based on the cross-sectional survey data of adult hypertension prevalence in Hunan Province from June 2013 to May 2014, 4 012 subjects aged ≥18 years with complete key data were included. Binary logistic regression analysis was used to analyze the correlation between TyG-BMI and hypertension, and receiver operating characteristic (ROC) curve was used to compare the ability of TyG index and TyG-BMI to identify hypertension.Results:A total of 4 012 subjects were included in this study, with an average age of (54.6±12.6)years, and males accounted for 40.98%(1 644/4 012). The prevalence of hypertension was 38.33%(1 538/4 012). Logistic regression analysis showed that elevated TyG-BMI was an independent risk factor for hypertension ( P<0.05). In the fully adjusted model, each 1/4 increase in TyG-BMI was associated with a 1.017-fold increase in the risk of hypertension ( OR=1.017, 95% CI: 1.014-1.019). Compared with the lowest quartile group (Q 1), the higher quartile groups (Q 2, Q 3, Q 4) of TyG-BMI had a higher risk of hypertension, with OR values of 1.841, 2.265, and 4.386, respectively. Restricted cubic spline plot showed a linear dose-response relationship between TyG-BMI and the risk of hypertension (overall trend P<0.001). In subgroup analyses stratified by age, gender, smoking and drinking status, TyG-BMI was positively correlated with hypertension. In addition, ROC curve analysis showed that TyG-BMI had better diagnostic value for hypertension compared with TyG index. Conclusions:TyG-BMI is an independent risk factor for hypertension. Excessively high TyG-BMI or gradual increase of TyG-BMI will increase the risk of hypertension, and TyG-BMI has higher value in identifying hypertension compared with TyG index.

Objective:To analyze the antimicrobial resistance and genomic characteristics of Salmonella typhimurium and its monophasic variants isolated from human and food sources in Hangzhou. Methods:A total of 186 Salmonella typhimurium and its monophasic variants isolated in Hangzhou during the period from 2017 to 2021 were subjected to antimicrobial susceptibility testing and whole-genome sequencing, and the genomes from public databases were downloaded. Multilocus sequence typing and the identification of antimicrobial resistance (AMR) genes were performed using the sequencing data. Phylogenetic tree based on the single nucleotide polymorphism (SNP) sites in the genomes from Hangzhou and public databases was constructed. Results:The multidrug resistance (MDR) rate of the monophasic variant strains in Hangzhou was 95.4% (103/108). The MDR strains harbored ≥3 classes of AMR genes simultaneously and the most common resistance genotype was blaTEM-1B- tet( B)- sul2- aac(6′)- Ia (44.7%, 46/103). The MDR rate of Salmonella typhimurium strains was 75.6% (59/78), and 52.5% (31/59) of the MDR strains harbored ≥10 AMR genes simultaneously, with the most common resistance genotype being blaTEM-1B- qnrS1- tet( A)- tet( M)- cmlA1- floR- dfrA12- sul2- sul3- aac(6′)- Iaa (40.7%, 24/59). All of the 186 strains were positive for AMR gene aac (6 ′)- Iaa, and three monophasic variant strains which were resistant to polymyxin E was positive for mcr-1.1. ST19 accounted for 91.0% of the 78 Salmonella typhimurium strains (except for two strains belonging to ST34, two strains belonging to ST1544, and three strains untyped), and ST34 accounted for 96.3% of the 108 monophasic variant strains (except for four strains untyped). The phylogenetic tree showed that the ST19 strains dominated by serotype typhimurium and the ST34 strains dominated by monophasic variants were divided into two clades. In the ST19 clade, strains from Hangzhou were mainly clustered with the strains from other cities in Zhejiang Province as well as other provinces and cities in China, such as Jiangxi, Shanghai, and Shandong. Strains isolated from pig, duck, and chicken were the ones that were clustered with clinical strains. In the ST34 clade, most strains from Hangzhou were clustered together, and some strains were clustered with those isolated from other cities in Zhejiang Province as well as Beijing, Guangdong, Shanghai, etc. Strains isolated from pig and chicken were the ones that were clustered with clinical strains. Conclusions:The MDR rate of monophasic variant strains is higher than that of Salmonella typhimurium strains, and the latter is prominent by its harboring multiple AMR genes. The epidemic of Salmonella typhimurium and its monophasic variants in Hangzhou is mainly caused by the spread of ST19 and ST34 strains, respectively, and the clinical infection may be closely related to the consumption of pork and poultry meat. While transmitting locally, cross-region transmission of the two serovars in Zhejiang Province and cross-province transmission in China may also occur.

Objective:To analyze the antimicrobial resistance and genomic characteristics of Salmonella typhimurium and its monophasic variants isolated from human and food sources in Hangzhou. Methods:A total of 186 Salmonella typhimurium and its monophasic variants isolated in Hangzhou during the period from 2017 to 2021 were subjected to antimicrobial susceptibility testing and whole-genome sequencing, and the genomes from public databases were downloaded. Multilocus sequence typing and the identification of antimicrobial resistance (AMR) genes were performed using the sequencing data. Phylogenetic tree based on the single nucleotide polymorphism (SNP) sites in the genomes from Hangzhou and public databases was constructed. Results:The multidrug resistance (MDR) rate of the monophasic variant strains in Hangzhou was 95.4% (103/108). The MDR strains harbored ≥3 classes of AMR genes simultaneously and the most common resistance genotype was blaTEM-1B- tet( B)- sul2- aac(6′)- Ia (44.7%, 46/103). The MDR rate of Salmonella typhimurium strains was 75.6% (59/78), and 52.5% (31/59) of the MDR strains harbored ≥10 AMR genes simultaneously, with the most common resistance genotype being blaTEM-1B- qnrS1- tet( A)- tet( M)- cmlA1- floR- dfrA12- sul2- sul3- aac(6′)- Iaa (40.7%, 24/59). All of the 186 strains were positive for AMR gene aac (6 ′)- Iaa, and three monophasic variant strains which were resistant to polymyxin E was positive for mcr-1.1. ST19 accounted for 91.0% of the 78 Salmonella typhimurium strains (except for two strains belonging to ST34, two strains belonging to ST1544, and three strains untyped), and ST34 accounted for 96.3% of the 108 monophasic variant strains (except for four strains untyped). The phylogenetic tree showed that the ST19 strains dominated by serotype typhimurium and the ST34 strains dominated by monophasic variants were divided into two clades. In the ST19 clade, strains from Hangzhou were mainly clustered with the strains from other cities in Zhejiang Province as well as other provinces and cities in China, such as Jiangxi, Shanghai, and Shandong. Strains isolated from pig, duck, and chicken were the ones that were clustered with clinical strains. In the ST34 clade, most strains from Hangzhou were clustered together, and some strains were clustered with those isolated from other cities in Zhejiang Province as well as Beijing, Guangdong, Shanghai, etc. Strains isolated from pig and chicken were the ones that were clustered with clinical strains. Conclusions:The MDR rate of monophasic variant strains is higher than that of Salmonella typhimurium strains, and the latter is prominent by its harboring multiple AMR genes. The epidemic of Salmonella typhimurium and its monophasic variants in Hangzhou is mainly caused by the spread of ST19 and ST34 strains, respectively, and the clinical infection may be closely related to the consumption of pork and poultry meat. While transmitting locally, cross-region transmission of the two serovars in Zhejiang Province and cross-province transmission in China may also occur.

Objective:To evaluate the immunogenicity and protective effect of a multicomponent recombinant protein vaccine EPRHP014 constructed independently and provide a scientific basis for developing new tuberculosis (TB) vaccine and effective prevention and control of TB.Methods:Three full-length Mycobacterium ( M.) tuberculosis protein antigens (EsxH, Rv2628, and HspX) and two epitope-predicted and optimized epitope-dominant protein antigens (nPPE18 and nPstS1) were selected, from which five protein antigens were used to construct a protein antigen composition EPRHP014, including a fusion expression multi-component protein antigen (EPRHP014f) and a multi-component mixed protein antigen (EPRHP014m) formed with the five single protein using clone, purification, and purification respectively. Multicomponent protein vaccines EPRHP014f and EPRHP014m were prepared with aluminum adjuvant, and the BCG vaccine was used as a control. ELISA detected the titer of serum-specific antibodies, the secretion of various cytokines was detected by ELISpot and Luminex, and immune protection was observed by the M.tuberculosis growth inhibition test in vitro. The results were statistically analyzed by t-test or rank sum test, and P<0.05 was considered a statistically significant difference. Results:Mice Immunized with EPRHP014m and EPRHP014f could produce highly effective IgG antibodies and their subtypes IgG1 and IgG2a, and the antibody titers were similar to those of mice immunized with BCG, with no statistical significance ( P>0.05). The number of spot-forming cells (SFC) secreting IFN-γ and IL-4 induced by EPRHP014f group was significantly higher than those by EPRHP014m group and BCG group ( P<0.05), but there was no significant difference in the number of SFC for IFN-γ and IL-4 induced between EPRHP014m group and BCG group ( P>0.05). The secretion levels of GM-CSF and IL-12p70 induced by the EPRHP014m group were higher than those of the BCG group ( P<0.05), but there was no significant difference in the levels of IL-6 and IL-10 induced between EPRHP014m group and BCG group ( P>0.05). There was no significant difference in the secretions of IL-6, IL-10, IL-12, and GM-CSF between the EPRHP014f and BCG groups ( P>0.05). EPRHP014m group, EPRHP014f group, and BCG group had obvious antibacterial effects in vitro, and the difference was insignificant ( P>0.05). Conclusion:Both EPRHP014f and EPRHP014m can induce strong humoral and cellular immune responses in mice after immunization, and have a strong ability to inhibit the growth of M. tuberculosis in vitro, indicating that the antigen composition EPRHP014 has good potential in the development and application of TB vaccine.

Objective:To analyze the antimicrobial resistance and genomic characteristics of Salmonella enterica serovar Derby strains isolated from human and food sources in Hangzhou. Methods:A total of 60 Salmonella enterica serovar Derby strains isolated in Hangzhou during the period from 2015 to 2020 were subjected to antimicrobial susceptibility testing, pulsed field gel electrophoresis (PFGE) typing and whole-genome sequencing. Multilocus sequence typing (MLST), core genome multilocus sequence typing (cgMLST) and the identification of antimicrobial resistance genes were performed using the sequencing data. Phylogenetic tree based on the single nucleotide polymorphism (SNP) sites in the 60 genomes from Hangzhou and 379 genomes from public databases was constructed. Results:No significant difference was observed in the drug resistance rates between the clinical strains and food strains in Hangzhou. The multidrug resistance (MDR) rate was 76.7% (46/60). All of the 60 Salmonella Derby strains were positive for the antimicrobial resistance genes aac(6′)- Iaa and fosA7. The 60 strains were subtyped into 46 molecular types by PFGE and 53 molecular types by cgMLST(HC2). Except for one strain belonging to ST3220, the other Salmonella Derby strains were ST40. The phylogenetic analysis showed that some strains isolated in Hangzhou were close to the strains in Southeast Asia, suggesting the possibility of cross-border transmission of ST40 strains, with the main food sources being pork and fish; other strains were close to those circulating in Beijing, Guangzhou, Hubei, Chongqing and other provinces, suggesting the possibility of cross-province transmission of the strains, with the main food sources being pork, beef and chicken. Conclusions:The epidemic of Salmonella Derby in Hangzhou was mainly caused by the spread of ST40 strains and MDR was common. Clinical infections might be closely related to the consumption of pork, beef, chicken and fish. There was the possibility of cross-border transmission of Salmonella Derby between Hangzhou and Southeast Asia and cross-province transmission in China.

Objective:To preliminarily evaluate the immunogenicity and efficacy of two novel tuberculosis vaccine candidates (a fusion multicomponent protein EPDPA015f and a mixed multicomponent protein EPDPA015m) and to provide a new antigen combination for the development of tuberculosis vaccines.Methods:Recombinant plasmids for the expression of EPDPA015f and EPDPA015m proteins were constructed. Six-week-old BALB/c mice were immunized with EPDPA015f or EPDPA015m in combination with aluminium adjuvant (50 μg/mouse) for three times with an interval of 10 d. The mice were sacrificed 10 d after the last immunization to collect blood and spleen samples. Serum antibody titers and cytokine levels were measured by ELISA, Luminex technique and enzyme-linked immunospot assay (ELISPOT). Mycobacterial growth inhibition assay (MGIA) was used to detect the ability of mouse splenocytes to inhibit the growth of Mtb in vitro. One-way analysis of variance and t-test were used for statistical analysis. Results:Both EPDPA015f and EPDPA015m could induce the production of various cytokines and IgG antibodies at a high level. The levels of cytokines related to Th1 (IL-2, TNF-α, IFN-γ), Th2 (IL-4, IL-6, IL-10) and Th17 (IL-17) as well as other proinflammatory cytokines (GM-CSF, IL-12) were higher in the EPDPA015f group than in the adjuvant group ( P<0.05). The titer of IgG antibody induced by EPDPA015f was as high as 1∶4×10 6. The results of MGIA showed that the numbers of Mtb (lgCFU) in the PBS, adjuvant, EPDPA015f and EPDPA015m groups were 3.46±0.11, 3.51±0.06, 2.98±0.09 and 3.19±0.08, respectively. The number of colonies in the EPDPA015f group was the least as compared with that in the other three groups ( P<0.001, P<0.001, P<0.01). Conclusions:The vaccine candidate EPDPA015f could elicit more comprehensive and high-level cellular and humoral immune responses, and exhibited superior in vitro inhibitory activity against the growth of Mtb. EPDPA015f had the potential to be used as a preventive vaccine or a booster vaccine

Objective:To investigate the regulation of transcription factor CCCTC binding factor (CTCF) on the expression of B-cell lymphoma 2 ( Bcl-2) gene in pterygium and its molecular mechanism. Methods:Pterygium tissue samples from 22 primary pterygium patients who underwent pterygium excision combined with autologous limbal stem cell transplantation in The First Hospital of Changsha from June 2017 to February 2019 were collected during the operation as pterygium group.Normal conjunctival tissue from 20 patients with ocular trauma due to conjunctiva rupture, eyeball rupture or eyeball perforation in the same period were collected during the repair of ocular trauma as control group.Real-time PCR and Western blot were used to detect the expression levels of CTCF and Bcl-2 in the two groups.The DNA methylation level of the Bcl-2 promoter in the samples of the two groups was detected by bisulfite sequencing PCR (BSP). Pterygium fibroblasts were isolated and cultured.Fibroblasts were identified by immunohistochemistry using vimentin antibody.The cultured pterygium fibroblasts were divided into a CTCF interference group transfected with CTCF interference plasmid, and a control group transfected with control plasmid.The expression levels of CTCF and Bcl-2 in pterygium fibroblasts in CTCF interference and control groups were detected by real-time PCR and Western blot.The cell vitality was detected with cell counting kit-8 at 12, 24, and 48 hours after transfection.The DNA methylation level of the Bcl-2 promoter in the cells of the CTCF interference and control groups after transfection was determined by BSP.Differences of the indexes among groups were analyzed.Correlation between Bcl-2 mRNA and Bcl-2 gene promoter methylation level of CTCF protein in pterygium tissue was analyzed by Pearson linear correlation analysis.This study protocol was approved by the Ethics Committee of The First Hospital of Changsha (No.KL-2017021). Written informed consent was obtained from the patients from whom the specimens were collected.Results:The relative expression levels of CTCF mRNA and protein in pterygium group were 7.23±3.34 and 0.92±0.21, respectively, which were significantly higher than 1.10±0.44 and 0.28±0.07 in normal conjunctiva group ( t=-8.136, -13.025; both at P<0.01). The relative expression levels of Bcl-2 mRNA and protein in pterygium group were 10.27±4.64 and 0.95±0.27, which were higher than 1.10±0.41 and 0.32±0.14 in normal conjunctiva group, showing statistically significant differences ( t=-8.789, -10.782; both at P<0.01). The CTCF protein expression was significantly positively correlated with the Bcl-2 mRNA expression in pterygium group ( r=0.746, P<0.01). The DNA methylation level of the Bcl-2 promoter in pterygium group was 0.65±0.09, which was lower than 0.83±0.06 in normal conjunctiva group, with a statistically significant difference ( t=7.408, P<0.01). The DNA methylation level was significantly negatively correlated with the Bcl-2 mRNA expression in pterygium group ( r=-0.635, P<0.01). After the interference of CTCF expression in pterygium fibroblasts, the relative expression levels of CTCF and Bcl-2 mRNA in CTCF interference group were 0.37±0.03 and 0.53±0.06, which were significantly lower than 1.02±0.06 and 0.99±0.07 in control group ( t=20.035, 9.029; both at P<0.01). The relative expression levels of CTCF and Bcl-2 proteins in CTCF interference group were 0.23±0.06 and 0.56±0.07, which were lower than 0.52±0.05 and 0.92±0.12 in control group, showing statistically significant differences ( t=6.914, 4.719; both at P<0.01). The cell viability of pterygium fibroblasts in CTCF interference group was 0.10±0.01, 0.17±0.01, 0.38±0.04 at 12, 24, and 48 hours after interference, respectively, which were lower than 0.12±0.01, 0.29±0.01 and 0.85±0.06 in control group, and the differences were statistically significant ( t=3.718, 18.350, 15.621; all at P<0.01). The DNA methylation level of Bcl-2 promoter in CTCF interference group was 0.75±0.04, which was significantly higher than 0.61±0.03 in control group ( t=-4.472, P<0.05). Conclusions:CTCF is excessively expressed in pterygium, which may mediate the overexpression of Bcl-2 through down-regulating DNA methylation level.

Objective:To investigate any anti-aging effect of repeated transcranial magnetic stimulation (rTMS) and explore the relationship between the effect and relief of clinical symptoms in patients with Parkinson′s disease (PD).Methods:A total of 108 PD patients were randomly divided into an rTMS group and a control group, each of 54, while another 54 healthy counterparts were selected to form a normal group. In addition to anti-PD drug therapy, the rTMS group was given daily rTMS treatment, 5 days a week for 4 weeks, while the control group received sham rTMS treatment, with no treatment of the normal group. Before the treatment and after 4 weeks of treatment as well as and 1 month after the ending of the treatment, the subjects′ clinical exercise symptoms were evaluated using the Unified Parkinson′s Disease Rating Scale (UPDRS), a timed exercise test and the 10m re-entry exercise test. Non-exercise symptoms were assessed using the Hamilton Depression Scale (HAMD), the Hamilton Anxiety Scale (HAMA) and the Mini-mental State Examination (MMSE). Fasting venous blood samples were analyzed to quantify the serum levels of tumor necrosis factor (TNF), interleukin-6 (IL-6), interleukin-1β (IL-1β) and matrix metalloproteinase-3 (MMP-3).Results:Four weeks and 1 month after the treatment, the average UPDRS scores, exercise test times and 10m re-entry exercise test results of the rTMS group were significantly better than those before treatment and significantly better than those of the control group at the same time point. The rTMS group′s average HAMA, HAMD and MMSE scores, as well as its average P300 latency and amplitude were also significantly better than those of the control group at the same time point and significantly better than those before treatment. After 4 weeks, the average MMP-3 content in the rTMS group was significantly lower than the control group′s average, and after a month the average levels of TNF, IL-6, IL-1β and MMP-3 of the rTMS group were all significantly different from those before treatment and those of the control group. The TNF, IL-6, IL-1β and MMP-3 levels were all positively correlated with the average UPDRS total score.Conclusion:High-frequency rTMS therapy can change the phenotypes related to cell senescence, and thus has good therapeutic effect on motor and non-motor symptoms of PD.