Objective To investigate the influence of repeated open field tests， novel object recognition tests， and Barnes maze behavioral tests within a short term on cognitive and anxiety assessment in 3xTg-AD mice.Methods Four groups of 3xTg-AD mice， aged 12 months， were obtained using different pretreatment regimens， and the mice in the experimental groups 1-3 were treated with different drugs， while those in the control group were treated with normal saline. The open field test， the novel object recognition test， and the Barnes maze test were performed on mice successively. A behavioral video analysis system was used to record the locomotor trajectories of the mice and analyze the parameters such as time spent in the central area， exploration time for novel versus familiar objects， and latency to reach the target hole. After one session of complete tests， three sessions were performed repeatedly， and all tests were completed within one month.Results With the increase in the number of repeated tests， there was a significant reduction in the time spent in the central area in the open field test （P<0.05）； in the novel object recognition test， there was an increase in the coefficient of variation for object recognition index and a reduction in exploratory behavior towards both the new environment and the novel objects； in the Barnes maze test， there was no significant difference in the latency to reach the target hole across the four tests， suggesting good reproducibility.Conclusion Repeated tests within a short term can interfere with the evaluation of anxiety status in 3xTg-AD mice in the open field test and their preference for novel objects in the novel object recognition test， but it has no obvious influence on the latency to reach the target hole in the Barnes maze test. Whether behavioral tests can be performed repeatedly in 3xTg-AD mice should be determined based on different tests.

Objective To explore the effect of hypoxia-inducible factor-1α(HIF-1α)inhibitor LW6 on ferroptosis in septic cardiomyopathy rats.Methods Rat septic cardiomyopathy model was prepared using cecal ligation and puncture(CLP)method.Thirty-six specific pathogen-free(SPF)6-8 weeks male SD rats were randomly divided into the sham-operated group,CLP group,CLP+solvent group,LW6 group,ferrostatin-1(Fer-1)group,and LW6+Fer-1 group.The degree of myocardial damage in each group was evaluated through hematoxylin-eosin stai-ning and detection of lactate dehydrogenase and creatine kinase content in cardiac tissue.Myocardial mitochondrial damage was observed by transmission electron microscopy.Ferroptosis level was determined by detecting iron ion concentration,reduced glutathione,malondialdehyde,and reactive oxygen species.Protein expression levels of HIF-1α,solute carrier family 7 member 11(SLC7A11),and glutathione peroxidase 4(GPX4)in cardiac tissue were detected by Western blotting.Results Compared with the CLP group and the CLP+solvent group,the LW6 group could ameliorate myocardial damage,alleviate mitochondrial damage,inhibit ferroptosis-related indicators(all P＜0.05),reduce HIF-1α protein expression levels(P＜0.05),and enhance SLC7A11 and GPX4 protein expression levels(both P＜0.05).Conclusion LW6 decreases HIF-1α expression and ferroptosis levels through the SLC7A11/GPX4 pathway,and ameliorates sepsis-induced cardiomyopathy.

Background and Aims:Early-onset extensive emphysematous pancreatitis(EP)is a rare but highly lethal subtype of infected pancreatic necrosis(IPN),characterized by abrupt onset and rapid deterioration.This study aimed to investigate its clinical characteristics,microbiological spectrum,treatment approaches,and outcomes to provide evidence for early identification and timely intervention.Methods:A retrospective analysis was performed on 305 IPN patients treated at Xiangya Hospital,Central South University,from January 2010 to October 2023.Eight patients who developed gas accumulation involving≥50%of pancreatic or peripancreatic necrosis within two weeks of onset were defined as early-onset extensive EP.Their clinical data were compared with those of ordinary IPN patients.Results:Early-onset extensive EP accounted for 2.6%of all IPN cases.The early-onset extensive EP group had significantly higher mortality and multiple organ failure rates compared with the ordinary IPN group(75.0%vs.24.6%and 75.0%vs.34.7%,respectively;both P<0.05).A total of 15 microbial isolates were identified from early-onset extensive EP patients,predominantly Klebsiella pneumoniae(62.5%)and Escherichia coli(37.5%).The infection rate of carbapenem-resistant Enterobacteriaceae(CRE)was markedly higher in the EP group than in the ordinary IPN group(75.0%vs.31.1%,P=0.015).Most patients were treated using a step-up approach based on percutaneous catheter drainage,with no significant difference in treatment strategy between the two groups(P=0.625).Conclusion:Early-onset extensive EP represents a rare and fulminant subtype of IPN with extremely poor outcomes.Klebsiella pneumoniae and CRE are the predominant pathogens.Early radiological evaluation and timely intervention are crucial for improving prognosis in these patients.

Background and Aims:Early-onset extensive emphysematous pancreatitis(EP)is a rare but highly lethal subtype of infected pancreatic necrosis(IPN),characterized by abrupt onset and rapid deterioration.This study aimed to investigate its clinical characteristics,microbiological spectrum,treatment approaches,and outcomes to provide evidence for early identification and timely intervention.Methods:A retrospective analysis was performed on 305 IPN patients treated at Xiangya Hospital,Central South University,from January 2010 to October 2023.Eight patients who developed gas accumulation involving≥50%of pancreatic or peripancreatic necrosis within two weeks of onset were defined as early-onset extensive EP.Their clinical data were compared with those of ordinary IPN patients.Results:Early-onset extensive EP accounted for 2.6%of all IPN cases.The early-onset extensive EP group had significantly higher mortality and multiple organ failure rates compared with the ordinary IPN group(75.0%vs.24.6%and 75.0%vs.34.7%,respectively;both P<0.05).A total of 15 microbial isolates were identified from early-onset extensive EP patients,predominantly Klebsiella pneumoniae(62.5%)and Escherichia coli(37.5%).The infection rate of carbapenem-resistant Enterobacteriaceae(CRE)was markedly higher in the EP group than in the ordinary IPN group(75.0%vs.31.1%,P=0.015).Most patients were treated using a step-up approach based on percutaneous catheter drainage,with no significant difference in treatment strategy between the two groups(P=0.625).Conclusion:Early-onset extensive EP represents a rare and fulminant subtype of IPN with extremely poor outcomes.Klebsiella pneumoniae and CRE are the predominant pathogens.Early radiological evaluation and timely intervention are crucial for improving prognosis in these patients.

OBJECTIVE To investigate the current status and existing issues of hospital-associated infection manage-ment in non-public medical institutions,so as to provide data for the formulation of hospital-associated infec-tion management policies and the construction of a quality evaluation system for hospital-associated infection man-agement in non-public medical institutions.METHODS A qualitative research method was employed.In Dec.2023,semi-structured interviews were conducted with 38 directors of hospital-associated infection management depart-ments in primary,secondary and tertiary non-public medical institutions.The interview data were coded and ana-lyzed by Nvivo 12.0 software to refine themes.RESULTS Three themes were refined:the organizational system for hospital-associated infection management in non-public medical institutions had been basically established,but the division of functions and responsibilities was not yet fully clear,hospital-associated infection management had achieved certain results,including collaborating with other departments to standardize and implement infection control measures,enhancing staff awareness of infection control,and promoting the establishment of organization-al systems.There were issues in infection control works such as insufficient funding,personnel allocation not meeting requirements,and difficulties in implementing infection control measures.The most urgent assis-tance needed was to address the training and professional title promotion for full-time and part-time infection con-trol personnel.CONCLUSION Non-public medical institutions should strengthen staffing and training,enhance the pro-fessional capabilities of infection control personnel,and establish hospital-associated infection management standards that are tailored to the characteristics of non-public medical institutions at all levels.

OBJECTIVE To investigate the current status and existing issues of hospital-associated infection manage-ment in non-public medical institutions,so as to provide data for the formulation of hospital-associated infec-tion management policies and the construction of a quality evaluation system for hospital-associated infection man-agement in non-public medical institutions.METHODS A qualitative research method was employed.In Dec.2023,semi-structured interviews were conducted with 38 directors of hospital-associated infection management depart-ments in primary,secondary and tertiary non-public medical institutions.The interview data were coded and ana-lyzed by Nvivo 12.0 software to refine themes.RESULTS Three themes were refined:the organizational system for hospital-associated infection management in non-public medical institutions had been basically established,but the division of functions and responsibilities was not yet fully clear,hospital-associated infection management had achieved certain results,including collaborating with other departments to standardize and implement infection control measures,enhancing staff awareness of infection control,and promoting the establishment of organization-al systems.There were issues in infection control works such as insufficient funding,personnel allocation not meeting requirements,and difficulties in implementing infection control measures.The most urgent assis-tance needed was to address the training and professional title promotion for full-time and part-time infection con-trol personnel.CONCLUSION Non-public medical institutions should strengthen staffing and training,enhance the pro-fessional capabilities of infection control personnel,and establish hospital-associated infection management standards that are tailored to the characteristics of non-public medical institutions at all levels.

Objective To explore the effect of hypoxia-inducible factor-1α(HIF-1α)inhibitor LW6 on ferroptosis in septic cardiomyopathy rats.Methods Rat septic cardiomyopathy model was prepared using cecal ligation and puncture(CLP)method.Thirty-six specific pathogen-free(SPF)6-8 weeks male SD rats were randomly divided into the sham-operated group,CLP group,CLP+solvent group,LW6 group,ferrostatin-1(Fer-1)group,and LW6+Fer-1 group.The degree of myocardial damage in each group was evaluated through hematoxylin-eosin stai-ning and detection of lactate dehydrogenase and creatine kinase content in cardiac tissue.Myocardial mitochondrial damage was observed by transmission electron microscopy.Ferroptosis level was determined by detecting iron ion concentration,reduced glutathione,malondialdehyde,and reactive oxygen species.Protein expression levels of HIF-1α,solute carrier family 7 member 11(SLC7A11),and glutathione peroxidase 4(GPX4)in cardiac tissue were detected by Western blotting.Results Compared with the CLP group and the CLP+solvent group,the LW6 group could ameliorate myocardial damage,alleviate mitochondrial damage,inhibit ferroptosis-related indicators(all P＜0.05),reduce HIF-1α protein expression levels(P＜0.05),and enhance SLC7A11 and GPX4 protein expression levels(both P＜0.05).Conclusion LW6 decreases HIF-1α expression and ferroptosis levels through the SLC7A11/GPX4 pathway,and ameliorates sepsis-induced cardiomyopathy.

Objective:To investigate the application value of metagenomic next-genera-tion sequencing (mNGS) in pathogenic diagnosis of suspected infected severe acute pancreatitis (SAP).Methods:The prospective study was conducted. The clinical data of 25 patients with suspected infected SAP who were admitted to the Xiangya Hospital of Central South University from May to September 2023 were collected. Upper limb venous blood samples of all the patients were collected for both of mNGS and routine pathogen microbial culture. Observation indicators: (1) grouping situations of the enrolled patients; (2) comparison of the diagnostic efficiency of mNGS and routine pathogen microbial culture; (3) results of peripheral blood pathogen microbial testing and peri-pancreatic effusion microbial culture; (4) testing time and cost. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M( Q1, Q3). Count data were expressed as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. Results:(1) Grouping situations of the enrolled patients. A total of 25 patients were selected for eligibility. There were 18 males and 7 females, aged 48(40,59)years. The duration of hospital stay of 25 patients was 30(20,50)days. The etiologies of 25 patients included 14 cases of hyperlipidemic pancreatitis, 8 cases of biliary pancreatitis, 1 case of alcohol-induced acute pancreatitis, and 2 cases of pancreatitis caused by other causes. Of the 25 patients, there were 17 cases with infected pancreatic necrosis (IPN) including 7 cases of death, and 8 cases with sterile pancreatic necrosis including no death. (2) Comparison of the diagnostic efficiency of mNGS and routine pathogen microbial culture. The positive rates of mNGS and routine pathogen microbial culture in diagnosis of suspected infected SAP were 72.0%(18/25) and 32.0%(8/25), respectively, showing a significant difference between them ( χ2=8.01, P<0.05). The sensitivity and negative predic-tive value of mNGS and routine pathogen microbial culture in diagnosis of IPN were 94.1%(16/17), 35.3%(6/17) and 85.7%(6/7), 35.3%(6/17), showing significant differences between them ( χ2=12.88, 5.04, P<0.05). The specificity and positive predictive value of mNGS and routine pathogen microbial culture in diagnosis of IPN were 75.0%(6/8), 75.0%(6/8) and 88.9%(16/18), 75.0%(6/8), showing no significant difference between them ( χ2=0, 0.82, P>0.05). (3) Results of peripheral blood pathogen microbial testing and peripancreatic effusion microbial culture. Of the 17 patients with IPN, 36 strains of pathogenic bacteria were detected by mNGS, and 6 strains were detected by routine pathogen microbial culture. There were 16 of 17 patients with IPN showing positive mNGS pathogenic testing, of which 13 cases were consistent with the pathogenic testing results of peri-pancreatic effusion microbial culture, showing a consistency rate of 76.5%(13/17). There were 6 pati-ents with IPN showing positive routine pathogen microbial culture, with a consistency rate of 35.3%(6/17) to peripancreatic effusion microbial culture. (4) Testing time and cost. Testing time of mNGS and routine pathogen microbial culture were (43±17)hours and (111±36)hours, showing a signifi-cant difference between them ( t=9.31, P<0.05). Testing cost of mNGS was (2 267±0)yuan/case, accoun-ting for 1.7% of the hospitalization expenses of (133 759±120 744)yuan/case. Testing cost of routine pathogen microbial culture was (240±0)yuan/case, accounting of 0.2% of the hospitalization expenses. Conclusion:mNGS has important value for early pathogenic diagnosis of suspected infected SAP, and has a high timeliness.

OBJECTIVE To establish a mouse model of diabetes mellitus(DM)combined with severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infection to investigate the important pathophysiological changes in the development of DM combined with SARS-CoV-2 infection.METHODS Wild-type(WT)mice and transgenic mice expressing the human angiotensin-converting enzyme 2 receptor driven by the cytokeratin-18 gene promoter(K18-hACE2)were randomly divided into the control group,DM group,SARS-CoV-2 spike protein(S)infection group and DM combined with S protein infection group,with 10 to 12 mice in each group.All the mice were induced by 10 weeks of high-fat diet combined with 40 mg·kg-1 streptozotocin(STZ)for 3 days by ip,except those in the control group or S protein infection group.The control group was given the same volume of 0.1 mol·L-1 sodium citrate buffer.Mice in the S protein infection group and DM+S protein infection group were additionally given 50 μL mixture of 15 μg SARS-CoV-2 spike protein and 1 g·L-1 polyinosinic-polycytidylic acid(poly[I:C])via intranasal drops,while the control group was given an equal volume of sterile water.The glucose tolerance level and pancreatic islet β cell function of mice were evaluated via oral glucose tolerance test at the 6th week of high-fat feeding and 1 week after the administration of STZ by ip.From the 6th week of high-fat feeding to 2 weeks after the administration of STZ,the random blood glucose and fasting blood glucose of mice were measured by a blood glucose meter.Blood samples were taken from subman-dibular veins of 3 mice in each group at 24,48 and 120 h after S protein infection,and lung tissues were taken after euthanization.The pathological changes of lungs of DM mice before and after S protein infection were observed by HE staining.Except for the DM group,blood samples were collected before S protein infection and at 6,24,48,72 and 120 h after infection.The levels of plasma interleukin 1β(IL-1β),IL-2,IL-6,IL-10,IL-17,interferon gamma-induced protein 10(IP-10),interferon γ(IFN-γ),tumor necrosis factor α(TNF-α),monocyte chemotactic protein-1(MCP-1)and granulocyte-colony stimulating factor(G-CSF)were detected by Luminex.The plasma levels of heparan sulfate(HS)were measured by enzyme-linked immunosorbent assay.The levels of cytokines and HS were correlated with the degree of pathological damage by Spearman correlation analysis.RESULTS STZ and high-fat diet could induce DM-like expression in mice,and the random blood glucose(P<0.01)and fasting blood glucose(P<0.05)after 1 week in the hACE2-DM group were significantly higher than in the WT-DM group,and the degree of islet function damage in hACE2-DM mice was significantly higher than that of WT-DM mice(P<0.05).Compared with the DM group,the DM+S group showed more severe pulmonary pathological changes after S protein infection,accompanied by a large number of inflammatory infiltrations and thickening of lung interstitial.Compared with the control group,the levels of pro-inflammatory cytokines G-CSF,IL-6 and IP-10 in the plasma of the WT-S group were significantly increased at 6 h after S pro-tein infection(P<0.01),and those of pro-inflammatory cytokine IL-17 and anti-inflammatory cytokine IL-10 were significantly increased at 24 h after S protein infection(P<0.05).Compared with the control group,the plasma levels of pro-inflammatory cytokines IL-1β,IL-6,TNF-α,MCP-1,G-CSF and IP-10 in the hACE2-S group were significantly increased at 6 h after S protein infection(P<0.05,P<0.01).IL-17 was significantly increased at 24 h and 6 h after S protein infection in the WT-DM+S group and hACE2-DM+S group,respectively(P<0.01,P<0.05).In the hACE2-DM+S group,IFN-γ and IL-1β were signifi-cantly increased in delay to 48 h(P<0.05,P<0.01),and MCP-1 was significantly increased in delay to 72h(P<0.05).Compared with the control group,the level of HS in the plasma of the WT-S group increased significantly(P<0.05,P<0.01)at 6 h and 24 h after S protein infection,but began to decrease at 48 h.At the same time,compared with the WT-S group,the HS level in the WT-DM+S group was slightly increased at 6 h after infection and decreased at 24 h.Compared with the control group,the HS level in the hACE2-S group was significantly increased at 24 h(P<0.01),as was the case with the WT-S group 24 h,48 h and 120 h after S protein infection.At 6 h,24 h and 48 h after S protein infection,the plasma HS level of the hACE2-DM+S group was significantly increased(P<0.01,P<0.05),and the duration of the increase was longer than in the hACE2-S group.Moreover,the levels of IL-1β,IL-10,MCP-1,IP-10,G-CSF and HS in plasma were positively correlated with the degree of lung dam-age in the DM+S group.CONCLUSION In this study,the mouse model of diabetes combined with SARS-CoV-2 spike protein infection has mimicked part of the pathophysiological features of clinical patients,mainly manifested as blunted immune response and elevated HS levels with longer duration to infection alone.IL-1β,IL-10,MCP-1,IP-10,G-CSF and HS may keep track of the course of disease in patients with diabetes combined with SARS-CoV-2 infection.

Objective:To establish an HPLC method for benzyl benzoate related substances and propose revision suggestions for the quality standard of this excipient.Methods:Phenylsilane bond and silica gel were used as fill-ers,and methanol-1％acetic acid(60:40)was used as the mobile phase at the flow rate of 1.0 mL·min-1.The detection wavelength was 254 nm and the injection amount was 20 μL.Results:Based on the results of the de-struction test and taking into account the separation requirements,the chromatographic conditions for benzyl benzo-ate related substances were confirmed and methodological validation was conducted.The detection limits for benzoic acid and benzaldehyde in related substances were 0.2 μg·mL-1 and 0.01 μg·mL-1.The correction factors for benzoic acid and benzaldehyde were 1.0 and 0.1,respectively.According to the newly established method,six batches of benzyl benzoate samples from four enterprises were inspected for related substances,and benzaldehyde was detected with content from 0.02％to 0.1％.The total amount of impurities was from 0.02％to 0.9％.Conclusion:The newly established method can be added to the quality standards of benzyl benzoate,and different impurity limits can be formulated according to the different administration routes in order to better control drug quality.