Objective To observe the effect of a Jianpi Huayu Jiedu formula on the NLRP3-mediated pyroptosis pathway in gastric precancerous lesion(GPL)associated with Helicobacter pylori(Hp)infection.Methods A GPL mouse model was prepared using Hp suspension gavage combined with Atp4a gene-deficient mice.The Jianpi Huayu Jiedu formula was administered as an intervention.Gastric mucosal tissue pathological damage was observed using hematoxylin and eosin(HE)staining.The presence of intestinal metaplasia(IM)was assessed using Alcian Blue-Periodic Acid Schiff(AB-PAS)staining.Ultrastructural changes in cell organelles were observed using transmission electron microscopy.Enzyme-linked immunosorbent assay(ELISA)was used to measure levels of gastrin-17(G-17),pepsinogen I(PGI),and proinflammatory cytokines IL-1β and IL-18.The expression of molecules related to the pyroptosis pathway was detected using Western blot and real-time quantitative PCR(RT-qPCR).Results Compared to the control group,Hp-related GPL mice exhibited gastric mucosal atrophy accompanied by IM and dysplasia.Damage to mitochondria and endoplasmic reticulum in parietal cells was observed.Levels of G-17,PGI,and proinflammatory cytokines IL-1β and IL-18 were elevated.The expression of molecules related to the pyroptosis pathway was increased.The Jianpi Huayu Jiedu formula significantly reduced gastric mucosal tissue pathological damage in GPL mice,decreased G-17 and PGI levels,mitigated inflammatory responses,and downregulated the expression of molecules related to the pyroptosis pathway.Conclusion The Jianpi Huayu Jiedu formula may exert its effects by inhibiting the NLRP3-mediated pyroptosis signaling pathway,thereby alleviating or even reversing the pathological damage of gastric mucosa in Hp-related GPL.

Objective:To investigate the role and molecular mechanisms of signal transducer and activator of tran-scription 3(STAT3)in the paraventricular nucleus(PVN)of the hypothalamus in regulating body weight and energy metabolism in mice.Methods:AAV2/9-hSyn-Cre-EGFP virus was stereotactically injected into the PVN of STAT3Flox/Flox mice to conditionally knock out(CKO)STAT3 in the PVN.STAT3 expression was verified via immunoflu-orescence staining and Western blot.Body weight and temperature were monitored,glucose tolerance was assessed using glucose tolerance tests in mice,and morphological changes in the liver,interscapular brown adipose tissue(IBAT),in-guinal white adipose tissue(IWAT),and quadriceps muscle were evaluated via hematoxylin-eosin(HE)staining.RT-qPCR was used to measure mRNA levels of platelet-derived growth factor receptor α(PDGFRα),peroxisome prolifera-tor-activated receptor γ(PPARγ),hormone-sensitive lipase(HSL),and adipose triglyceride lipase(ATGL)in these tissues.Results:STAT3 protein expression in the PVN of CKO mice was significantly reduced,and the number of STAT3-positive neurons was also decreased.Compared to wild-type(WT)mice,CKO mice exhibited increased body weight,impaired thermogenesis in IBAT,and reduced glucose tolerance.HE staining revealed lipid droplet accumula-tion in hepatocytes of the liver,enlarged adipocytes with hypertrophic lipid droplets and leukocyte infiltration in adipose tissues,and intermuscular fat deposition in the quadriceps muscle.RT-qPCR analysis showed decreased mRNA levels of PDGFRα,HSL,and ATGL in the liver;upregulated PPARγ mRNA but downregulated HSL and ATGL mRNA in IBAT and IWAT;and reduced PPARγ and HSL mRNA levels in the quadriceps muscle of CKO mice.Conclusion:The STAT3 signaling pathway in the PVN is critical for maintaining systemic energy balance and serves as a key node in met-abolic regulation.

Objective:To investigate the role and molecular mechanisms of signal transducer and activator of tran-scription 3(STAT3)in the paraventricular nucleus(PVN)of the hypothalamus in regulating body weight and energy metabolism in mice.Methods:AAV2/9-hSyn-Cre-EGFP virus was stereotactically injected into the PVN of STAT3Flox/Flox mice to conditionally knock out(CKO)STAT3 in the PVN.STAT3 expression was verified via immunoflu-orescence staining and Western blot.Body weight and temperature were monitored,glucose tolerance was assessed using glucose tolerance tests in mice,and morphological changes in the liver,interscapular brown adipose tissue(IBAT),in-guinal white adipose tissue(IWAT),and quadriceps muscle were evaluated via hematoxylin-eosin(HE)staining.RT-qPCR was used to measure mRNA levels of platelet-derived growth factor receptor α(PDGFRα),peroxisome prolifera-tor-activated receptor γ(PPARγ),hormone-sensitive lipase(HSL),and adipose triglyceride lipase(ATGL)in these tissues.Results:STAT3 protein expression in the PVN of CKO mice was significantly reduced,and the number of STAT3-positive neurons was also decreased.Compared to wild-type(WT)mice,CKO mice exhibited increased body weight,impaired thermogenesis in IBAT,and reduced glucose tolerance.HE staining revealed lipid droplet accumula-tion in hepatocytes of the liver,enlarged adipocytes with hypertrophic lipid droplets and leukocyte infiltration in adipose tissues,and intermuscular fat deposition in the quadriceps muscle.RT-qPCR analysis showed decreased mRNA levels of PDGFRα,HSL,and ATGL in the liver;upregulated PPARγ mRNA but downregulated HSL and ATGL mRNA in IBAT and IWAT;and reduced PPARγ and HSL mRNA levels in the quadriceps muscle of CKO mice.Conclusion:The STAT3 signaling pathway in the PVN is critical for maintaining systemic energy balance and serves as a key node in met-abolic regulation.

Objective To investigate the killing effect of the recombinant oncolytic influenza virus OvFlu-GM-CSF,constructed using reverse genetics(RG)technology,in combination with chemotherapy drug,gemcitabine(GEM),against pancreatic cancer cells.Methods The recombinant oncolytic virus OvFlu-GM-CSF was successfully rescued using RG technology in our previous study.The virus was then comprehensively characterized through chicken red blood cell hemagglutination assay,transmission electron microscopy,and viral replication assay.CCK-8 assay was utilized to determine the impact of OvFlu-GM-CSF viruses[multiplicities of infection(MOI):0,0.1,1.0,3.0]on the survival rate of pancreatic cancer cell lines(Panc02,PANC-1,SW1990,BxPC-3)and normal pancreatic ductal epithelial cells(HPDE6-C7)after treatment for 24,48 or 72 h.Using a subcutaneous tumor-bearing mouse model of pancreatic cancer,36 female C57BL/6 mice(6 weeks old)were randomly divided into PBS group,recombinant oncolytic virus group,GEM group,and the combined treatment group,with 9 mice in each group.PBS(100 μL/animal)or OvFlu-GM-CSF virus(1× 107PFU/100 μL)was given to the mice of the corresponding groups through intratumoral injection,while GEM(100 mg/kg)was injected intraperitoneally,once per 3 days,for totally 9 times.Changes in tumor volume and survival rate were monitored.Multi-immunofluorescence staining was employed to analyze T cell infiltration and proliferation in the tumor tissues.HE staining was performed to observe the pathological changes in major organs(heart,liver,lungs,kidneys and brain),and the serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were measured to evaluate the safety of the recombinant oncolytic virus.Results The recombinant oncolytic influenza virus OvFlu-GM-CSF has a hemagglutination titer of 28,typical morphological features of influenza virus,and can selectively replicate within pancreatic cancer cells.At the cellular level,the viruses demonstrated a significant selective cytotoxic effect on Panc02,PANC-1,SW1990,and BxPC-3 cells under the conditions of 48 h post-infection and MOI=3.0,when compared to 48 h post-infection and MOI=0(P＜0.01).The cell survival rate was gradually decreased with the increase in MOI value and the extension of infection time(P＜0.01),but the viruses showed no significant effect on normal pancreatic ductal epithelial cells(HPDE6-C7).In the pancreatic cancer tumor-bearing mouse model,the combined treatment of the viruses+GEM significantly reduced the tumor volume than simple virus treatment and simple GEM treatment(P＜0.01),and enhanced the infiltration of T cells in the tumor tissues.No obvious pathological changes were observed in the above-mentioned major organs.Additionally,there were no significant differences in the serum levels of ALT and AST in the OvFlu-GM-CSF group,GEM group,and OvFlu-GM-CSF+GEM group compared to the PBS group.Conclusion RS technology-constructed recombinant oncolytic influenza virus OvFlu-GM-CSF,when combined with the chemotherapeutic agent GEM,enhances the cytotoxic efficacy against pancreatic cancer cells and effectively activates the host's anti-tumor immune response.

Objective:To investigate the application of time series models in forecasting pre-hospital emergency ambulance trips in Handan City and develop the LPro ensemble model for improved prediction accuracy to support emergency resource allocation.Methods:Pre-hospital emergency data from Handan Emergency Medical Command Center (2019-2023) were retrospectively analyzed. From 324 799 original records, 289 949 valid records were included after cleaning. The training set (2019-2022: 215 918 records) included 35 527 records in 2019, 52 015 in 2020, 61 836 in 2021, and 66 540 in 2022. The validation set (2023) contained 74 031 records. ARIMA, linear trend seasonal, exponential smoothing, and Prophet models were fitted to the training set. The LPro ensemble model was constructed using MAPE-based weighting (linear trend seasonal model: 0.38, Prophet: 0.62). Performance metrics included MAPE, RMSE, MAE, and R 2. Results:Data showed annual growth (compound annual growth rate 23.27%) and seasonal patterns (October peaks, February troughs). Ambulance dispatches increased annually with monthly cyclical patterns. For 2023 validation predictions: ARIMA (MAPE 8.76%, RMSE 619, MAE 491, R 2 0.4563), linear trend seasonal (MAPE 9.83%, RMSE 671, MAE 545, R 2 0.3608), Prophet (MAPE 8.43%, RMSE 562, MAE 503, R 2 0.5513), exponential smoothing (MAPE 8.08%, RMSE 643, MAE 410, R 2 0.4124). LPro model showed superior performance (MAPE 7.05%, RMSE 491, MAE 393, R 2 0.6570), with 16.37% lower MAPE, 12.63% lower RMSE, 21.87% lower MAE, and 19.17% higher R 2 versus Prophet. Conclusion:The LPro ensemble model substantially enhances prediction accuracy and reliability, offering scientific support for emergency resource optimization and dispatch scheduling in Handan City.

Objective:To investigate the epidemiological and clinical characteristics of patients with nontuberculous mycobacterial infections in Hunan Province using metagenomics next generation sequencing (mNGS) .Methods:Pathological data form 320 patients diagnosed with mycobacterial infections at the Department of Pathology of Hunan People′s Hospital from June 2019 to December 2023 were collected, including HE staining, acid-fast staring, and fungal fluorescence staining. mNGS was utilized for the classification of mycobacterial pathogens.Results:Among 320 patients with mycobacterial infections, 249 cases were positive for Mycobacterium tuberculosis and 71 cases were positive for nontuberculous mycobacteria. Thirteen genera of nontuberculous mycobacteria were identified from the mycobacterial samples, including Mycobacterium chelonae, Mycobacterium avium complex, Mycobacterium abscessus, Mycobacterium gordonae, Mycobacterium intracellulare, Mycobacterium xenopi, Mycobacterium kansasii, Mycobacterium paragordonae, Mycobacterium obuense, Mycobacterium marinum, Mycobacterium fortuitum, Mycobacterium smegmatis and Mycobacterium mageritense. Conclusions:Infections caused by nontuberculous mycobacteria are gradually increasing, transitioning from single-strain infections to multi-strain complex infections. The histopathological features of nontuberculous mycobacterial infections are difficult to distinguish form those of tuberculosis, posing significant challenges for clinical diagnosis and treatment.

Objective To observe the effect of a Jianpi Huayu Jiedu formula on the NLRP3-mediated pyroptosis pathway in gastric precancerous lesion(GPL)associated with Helicobacter pylori(Hp)infection.Methods A GPL mouse model was prepared using Hp suspension gavage combined with Atp4a gene-deficient mice.The Jianpi Huayu Jiedu formula was administered as an intervention.Gastric mucosal tissue pathological damage was observed using hematoxylin and eosin(HE)staining.The presence of intestinal metaplasia(IM)was assessed using Alcian Blue-Periodic Acid Schiff(AB-PAS)staining.Ultrastructural changes in cell organelles were observed using transmission electron microscopy.Enzyme-linked immunosorbent assay(ELISA)was used to measure levels of gastrin-17(G-17),pepsinogen I(PGI),and proinflammatory cytokines IL-1β and IL-18.The expression of molecules related to the pyroptosis pathway was detected using Western blot and real-time quantitative PCR(RT-qPCR).Results Compared to the control group,Hp-related GPL mice exhibited gastric mucosal atrophy accompanied by IM and dysplasia.Damage to mitochondria and endoplasmic reticulum in parietal cells was observed.Levels of G-17,PGI,and proinflammatory cytokines IL-1β and IL-18 were elevated.The expression of molecules related to the pyroptosis pathway was increased.The Jianpi Huayu Jiedu formula significantly reduced gastric mucosal tissue pathological damage in GPL mice,decreased G-17 and PGI levels,mitigated inflammatory responses,and downregulated the expression of molecules related to the pyroptosis pathway.Conclusion The Jianpi Huayu Jiedu formula may exert its effects by inhibiting the NLRP3-mediated pyroptosis signaling pathway,thereby alleviating or even reversing the pathological damage of gastric mucosa in Hp-related GPL.

Objective:To investigate the epidemiological and clinical characteristics of patients with nontuberculous mycobacterial infections in Hunan Province using metagenomics next generation sequencing (mNGS) .Methods:Pathological data form 320 patients diagnosed with mycobacterial infections at the Department of Pathology of Hunan People′s Hospital from June 2019 to December 2023 were collected, including HE staining, acid-fast staring, and fungal fluorescence staining. mNGS was utilized for the classification of mycobacterial pathogens.Results:Among 320 patients with mycobacterial infections, 249 cases were positive for Mycobacterium tuberculosis and 71 cases were positive for nontuberculous mycobacteria. Thirteen genera of nontuberculous mycobacteria were identified from the mycobacterial samples, including Mycobacterium chelonae, Mycobacterium avium complex, Mycobacterium abscessus, Mycobacterium gordonae, Mycobacterium intracellulare, Mycobacterium xenopi, Mycobacterium kansasii, Mycobacterium paragordonae, Mycobacterium obuense, Mycobacterium marinum, Mycobacterium fortuitum, Mycobacterium smegmatis and Mycobacterium mageritense. Conclusions:Infections caused by nontuberculous mycobacteria are gradually increasing, transitioning from single-strain infections to multi-strain complex infections. The histopathological features of nontuberculous mycobacterial infections are difficult to distinguish form those of tuberculosis, posing significant challenges for clinical diagnosis and treatment.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in healthcare facilities in major regions of China in 2023.Methods Clinical isolates collected from 73 hospitals across China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2023 Clinical & Laboratory Standards Institute (CLSI) breakpoints.Results A total of 445199 clinical isolates were collected in 2023,of which 29.0％ were gram-positive and 71.0％ were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species (excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi) (MRSA,MRSE and MRCNS) was 29.6％,81.9％ and 78.5％,respectively.Methicillin-resistant strains showed significantly higher resistance rates to most antimicrobial agents than methicillin-susceptible strains (MSSA,MSSE and MSCNS).Overall,92.9％ of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 91.4％ of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis had significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 93.1％ in the isolates from children and and 95.9％ in the isolates from adults.The resistance rate to carbapenems was lower than 15.0％ for most Enterobacterales species except for Klebsiella,22.5％ and 23.6％ of which were resistant to imipenem and meropenem,respectively .Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.6％ to 10.0％.The resistance rate to imipenem and meropenem was 21.9％ and 17.4％ for Pseudomonas aeruginosa,respectively,and 67.5％ and 68.1％ for Acinetobacter baumannii,respectively.Conclusions Increasing resistance to the commonly used antimicrobial agents is still observed in clinical bacterial isolates.However,the prevalence of important crabapenem-resistant organisms such as crabapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a slightly decreasing trend.This finding suggests that strengthening bacterial resistance surveillance and multidisciplinary linkage are important for preventing the occurrence and development of bacterial resistance.