ObjectiveTo analyze the infection status of main respiratory pathogens in influenza-like illness (ILI) cases in Anji County, Huzhou City, Zhejiang Province, and to provide a reference for the prevention, diagnosis, and treatment of respiratory infections. MethodsThroat swab samples were collected from 520 ILI cases in an influenza sentinel surveillance hospital in Anji County of Zhejiang Province from December 2023 to November 2024. Multiplex real-time fluorescence quantitative polymerase chain reaction (mRT-PCR) was used to detect 18 pathogens and their subtypes, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza A virus (Flu A), influenza A (H1N1) virus, influenza A (H3N2) virus, influenza B virus (Flu B), influenza B virus Victoria lineage (BV), influenza B virus Yamagata lineage (BY), coronavirus (CoV), human parainfluenza virus (HPIV), respiratory syncytial virus (RSV), human metapneumovirus (HMPV), adenovirus (ADV), human bocavirus (HBoV), enterovirus (EV), rhinovirus (RV), Mycoplasma pneumoniae (MP), Chlamydia pneumoniae (CP), and Streptococcus pneumoniae (SP). ResultsThe overall positivity rate of pathogens in 520 samples was 33.65%, among which the detection rates of Flu (9.14%), ADV (7.50%), SARS-CoV-2 (6.15%), and EV (3.65%) were relatively high. There were statistically significant differences in the overall positivity rate of pathogens by age and season (all P<0.05). The highest overall positivity rate was observed in the 5‒14 years old group (42.77%), and the overall positivity rate in winter (53.08%) was significantly higher than that in other seasons. ConclusionFrom 2023 to 2024, the main respiratory pathogens detected in ILI cases in Anji County were Flu, ADV, SARS-CoV-2, and EV. The epidemic characteristics showed age and seasonal specificity, so it is necessary to strengthen prevention and control for high-risk populations and epidemic seasons in a targeted manner.

Objective:To construct a prediction model for gestational diabetes mellitus (GDM) complicated with reproductive tract infections based on the decision tree algorithm, and explore prevention and control strategies for GDM complicated with reproductive tract infections.Methods:This cross-sectional study used convenience sampling to select 256 GDM patients treated at the Obstetrics and Gynecology Hospital, School of Medicine, Zhejiang University from January 2023 to June 2024 as the study subjects. Based on the occurrence of reproductive tract infections, the patients were divided into a non-infection group and an infection group. Multivariate Logistic regression analysis and the decision tree method were employed to create a prediction model for GDM complicated with reproductive tract infections, and the risk prediction performance of the model was evaluated.Results:Among the 256 GDM patients, the age ranged from 20 to 43 (33.42 ± 4.25) years. The incidence of reproductive tract infections was 18.75% (48/256). Multivariate Logistic regression analysis revealed that age, glycosylated hemoglobin>6%, history of reproductive tract infections, uncontrolled blood glucose, and frequency of sexual activity ≥2 times during mid-to-late pregnancy were independent risk factors for reproductive tract infections in GDM patients ( χ2 values were 2.08-4.24, all P<0.05). The decision tree model for GDM complicated with reproductive tract infections consisted of 4 layers and 17 nodes, with the nodes selecting five indicators: age, frequency of sexual activity during mid-to-late pregnancy, blood glucose control status, sharing of vulvar cleaning tools, and history of reproductive tract infections. Among these, age was the most significant predictor of reproductive tract infections in GDM patients. The area under the curve (AUC) value of the decision tree model for GDM complicated with reproductive tract infections was 0.948, while the AUC value of the multivariate logistic regression model was 0.927. There was no statistically significant difference between the AUC values of the decision tree model and the multivariate logistic regression model ( P>0.05). Conclusions:The constructed decision tree model for GDM patients developing genital tract infections has high application value and can serve as a basis for clinical screening of potential GDM patients at risk of genital tract infections.

Objective To explore the correlation between the expression level of serum fibrinogen-like protein 1(FGL1)and the indexes of metabolism and renal function in patients with diabetic nephropathy(DN)and diabetes mellitus(DM),and provide reference for clinical diagnosis and treatment.Methods From January 2017 to April 2023,30 patients with DM and treated in Jiangsu Province Hospital of Chinese Medicine were selected as the DM group,68 patients with DN were selected as the DN group,and 36 healthy subjects were selected as the control group.The DN group was further divided into the early DN(DN-E)group(n=38)and the late DN(DN-A)group(n=30)according to whether there was a large amount of proteinuria and the severity.Clinical data such as serum albumin(ALB),estimated glomerular filtration rate(eGFR)and albumin-to-creatinine ratio(ACR)were collected.Serum FGL1 level was detected by enzyme-linked immunosorbent assay(ELISA).Pearson linear correlation was used for correlation,the diagnostic value was analyzed by ROC curve.Results Compared with the control group,the levels of ACR,FGL1 in patients with DM group increased,the levels of eGFR decreased,and the differences were statistically significant(t=5.686,4.336,-4.683,all P<0.05).Compated with the DM group,the levels of ACR,FGL1 in patients with DN-E group was increased,and the level of eGFR was decreased,and the differences were statistically significant(t=5.275,3.454,-4.969,all P<0.05).Compared with the DN-E group,the levels of ACR,FGL1 in the DN-A group were increased,the levels of eGFR were decreased,and the differences were statistically significant(t=7.881,7.051,-5.596,all P<0.05).Serum FGL1 level was negatively correlated with ALB and eGFR(r=-0.638,-0.547,all P<0.05),and positively correlated with ACR(r=0.691,P<0.05).The AUC(95%CI),specificity and sensitivity of serum FGL1 level in the diagnosis of DN were 0.947(0.908～0.987),100%and 82.4%,respectively.Conclusion The level of serum FGL1 in DN and DM patients is high,and the level of serum FGL1 is closely related to the common metabolic indexes such as ALB,eGFR and ACR in the diagnosis of DN,which may have certain clinical diagnostic value.

Objective To develop a gas chromatography-tandem mass spectrometry(GC-MS/MS)method for the simultaneous determination of ephedrine and pseudoephedrine in urine.Methods Urine samples containing ephedrine and pseudoephedrine components were extracted with ethyl acetate,centrifuged to collect the supernatant and evaporated to dryness under a nitrogen stream and then derivatized with heptafluorobutyric anhydride 60 μL at 70 ℃ for 30 min,and re-evaporated under nitrogen,and then solubilized with 50 μL of methanol,and then analyzed by GC-MS/MS.Results The method demonstraed excellent linearity for ephedrine(0.05～10 μg/mL,r=0.999 8)and pseudoephedrine(0.02～5 μg/mL,r=0.999 5).Extraction recoveries ranged from 89.4％～95.8％(ephedrine)and 90.3％～93.8％(pseudoephedrine).Limits of detection and quantification of ephedrine and pseudoephedrine were 0.005 μg/mL and 0.01 μg/mL,the intra-day precision and accuracy were less than 5.87％and 9.56％,respectively,and the inter-day precision and accuracy were less than 7.54％and 9.27％,respectively.The stability of ephedrine and pseudoephedrine in urine in 15 d was good under the conditions of room temperature and-20 ℃.Conclusion The GC-MS/MS analytical method for the analysis of ephedrine and pseudoephedrine components in urine established in this study is accurate,stable and sensitive,which can provide data technical support for the forensic toxicological analysis of amphetamine-type drugs or new psychoactive substances in the cathinone group.

Objective To develop a gas chromatography-tandem mass spectrometry(GC-MS/MS)method for the simultaneous determination of ephedrine and pseudoephedrine in urine.Methods Urine samples containing ephedrine and pseudoephedrine components were extracted with ethyl acetate,centrifuged to collect the supernatant and evaporated to dryness under a nitrogen stream and then derivatized with heptafluorobutyric anhydride 60 μL at 70 ℃ for 30 min,and re-evaporated under nitrogen,and then solubilized with 50 μL of methanol,and then analyzed by GC-MS/MS.Results The method demonstraed excellent linearity for ephedrine(0.05～10 μg/mL,r=0.999 8)and pseudoephedrine(0.02～5 μg/mL,r=0.999 5).Extraction recoveries ranged from 89.4％～95.8％(ephedrine)and 90.3％～93.8％(pseudoephedrine).Limits of detection and quantification of ephedrine and pseudoephedrine were 0.005 μg/mL and 0.01 μg/mL,the intra-day precision and accuracy were less than 5.87％and 9.56％,respectively,and the inter-day precision and accuracy were less than 7.54％and 9.27％,respectively.The stability of ephedrine and pseudoephedrine in urine in 15 d was good under the conditions of room temperature and-20 ℃.Conclusion The GC-MS/MS analytical method for the analysis of ephedrine and pseudoephedrine components in urine established in this study is accurate,stable and sensitive,which can provide data technical support for the forensic toxicological analysis of amphetamine-type drugs or new psychoactive substances in the cathinone group.

Objective To explore the correlation between the expression level of serum fibrinogen-like protein 1(FGL1)and the indexes of metabolism and renal function in patients with diabetic nephropathy(DN)and diabetes mellitus(DM),and provide reference for clinical diagnosis and treatment.Methods From January 2017 to April 2023,30 patients with DM and treated in Jiangsu Province Hospital of Chinese Medicine were selected as the DM group,68 patients with DN were selected as the DN group,and 36 healthy subjects were selected as the control group.The DN group was further divided into the early DN(DN-E)group(n=38)and the late DN(DN-A)group(n=30)according to whether there was a large amount of proteinuria and the severity.Clinical data such as serum albumin(ALB),estimated glomerular filtration rate(eGFR)and albumin-to-creatinine ratio(ACR)were collected.Serum FGL1 level was detected by enzyme-linked immunosorbent assay(ELISA).Pearson linear correlation was used for correlation,the diagnostic value was analyzed by ROC curve.Results Compared with the control group,the levels of ACR,FGL1 in patients with DM group increased,the levels of eGFR decreased,and the differences were statistically significant(t=5.686,4.336,-4.683,all P<0.05).Compated with the DM group,the levels of ACR,FGL1 in patients with DN-E group was increased,and the level of eGFR was decreased,and the differences were statistically significant(t=5.275,3.454,-4.969,all P<0.05).Compared with the DN-E group,the levels of ACR,FGL1 in the DN-A group were increased,the levels of eGFR were decreased,and the differences were statistically significant(t=7.881,7.051,-5.596,all P<0.05).Serum FGL1 level was negatively correlated with ALB and eGFR(r=-0.638,-0.547,all P<0.05),and positively correlated with ACR(r=0.691,P<0.05).The AUC(95%CI),specificity and sensitivity of serum FGL1 level in the diagnosis of DN were 0.947(0.908～0.987),100%and 82.4%,respectively.Conclusion The level of serum FGL1 in DN and DM patients is high,and the level of serum FGL1 is closely related to the common metabolic indexes such as ALB,eGFR and ACR in the diagnosis of DN,which may have certain clinical diagnostic value.

OBJECTIVES: To investigate the role of the BNIP3-PI3K/Akt signaling pathway in mediating the inhibitory effect of Buyang Huanwu Decoction (BYHWT) on mitochondrial autophagy in human synovial fibroblasts from rheumatoid arthritis patients (FLS-RA) cultured under a hypoxic condition. METHODS: Forty normal Wistar rats were randomized into two groups (n=20) for daily gavage of BYHWT or distilled water for 7 days to prepare BYHWT-medicated or control sera. FLS-RA were cultured in routine condition or exposed to hypoxia (10% O₂) for 24 h wigh subsequent treatment with IL-1β, followed by treatment with diluted BYHWT-medicated serum (5%, 10% and 20%) or control serum. AnnexinV-APC/7-AAD double staining and T-AOC kit were used for detecting apoptosis and total antioxidant capacity of the cells, and the changes in ROS, ATP level, mitochondrial membrane potential and Ca²⁺ homeostasis were analyzed. The changes in mRNA and protein expressions of BNIP3, PI3K and AKT and mRNA expressions of LC3, Beclin-1 and P62 were detected using RT-qPCR and Western blotting. RESULTS: Treatment with BYHWT-medicated serum dose-dependently lowered apoptosis rate of IL-1β-induced FLS-RA with hypoxic exposure. The treatment significantly decreased T-AOC concentration, increased ROS production, autophagosome formation and ATPase levels, and lowered mitochondrial membrane potential and Ca²⁺ level in the cells. In IL-1β-induced FLS-RA with hypoxic exposure, treatment with BYHWT-medicated serum significantly increased BNIP3 protein expression, decreased the protein expressions of PI3K and AKT, increased the mRNA expressions of BNIP3 and P62, and lowered the mRNA expressions of PI3K, AKT, LC3 and Beclin-1 without significantly affecting Beclin-1 protein expression. The cells treated with 5% and 10% BYHWT-medicated serum showed no significant changes in LC3 expression. CONCLUSIONS BYHWT inhibits mitochondrial autophagy in IL-1β-induced FLS-RA with hypoxic exposure possibly by inhibiting BNIP3-mediated PI3K/AKT signaling pathway.
Drugs, Chinese Herbal/pharmacology* ; Arthritis, Rheumatoid/pathology* ; Animals ; Signal Transduction/drug effects* ; Proto-Oncogene Proteins c-akt/metabolism* ; Autophagy/drug effects* ; Humans ; Fibroblasts/cytology* ; Rats, Wistar ; Membrane Proteins/metabolism* ; Rats ; Phosphatidylinositol 3-Kinases/metabolism* ; Mitochondria/metabolism* ; Cells, Cultured ; Proto-Oncogene Proteins/metabolism* ; Apoptosis/drug effects* ; Cell Hypoxia ; Synovial Membrane/cytology* ; Male ; Mitochondrial Proteins

Objective To investigate the role of the BNIP3-PI3K/Akt signaling pathway in mediating the inhibitory effect of Buyang Huanwu Decoction(BYHWT)on mitochondrial autophagy in human synovial fibroblasts from rheumatoid arthritis patients(FLS-RA)cultured under a hypoxic condition.Methods Forty normal Wistar rats were randomized into two groups(n=20)for daily gavage of BYHWT or distilled water for 7 days to prepare BYHWT-medicated or control sera.FLS-RA were cultured in routine condition or exposed to hypoxia(10％O2)for 24 h wigh subsequent treatment with IL-1β,followed by treatment with diluted BYHWT-medicated serum(5％,10％and 20％)or control serum.AnnexinV-APC/7-AAD double staining and T-AOC kit were used for detecting apoptosis and total antioxidant capacity of the cells,and the changes in ROS,ATP level,mitochondrial membrane potential and Ca2+homeostasis were analyzed.The changes in mRNA and protein expressions of BNIP3,PI3K and AKT and mRNA expressions of LC3,Beclin-1 and P62 were detected using RT-qPCR and Western blotting.Results Treatment with BYHWT-medicated serum dose-dependently lowered apoptosis rate of IL-1β-induced FLS-RA with hypoxic exposure.The treatment significantly decreased T-AOC concentration,increased ROS production,autophagosome formation and ATPase levels,and lowered mitochondrial membrane potential and Ca2+level in the cells.In IL-1β-induced FLS-RA with hypoxic exposure,treatment with BYHWT-medicated serum significantly increased BNIP3 protein expression,decreased the protein expressions of PI3K and AKT,increased the mRNA expressions of BNIP3 and P62,and lowered the mRNA expressions of PI3K,AKT,LC3 and Beclin-1 without significantly affecting Beclin-1 protein expression.The cells treated with 5％and 10％BYHWT-medicated serum showed no significant changes in LC3 expression.Conclusion BYHWT inhibits mitochondrial autophagy in IL-1β-induced FLS-RA with hypoxic exposure possibly by inhibiting BNIP3-mediated PI3K/AKT signaling pathway.

Objective:To construct a prediction model for gestational diabetes mellitus (GDM) complicated with reproductive tract infections based on the decision tree algorithm, and explore prevention and control strategies for GDM complicated with reproductive tract infections.Methods:This cross-sectional study used convenience sampling to select 256 GDM patients treated at the Obstetrics and Gynecology Hospital, School of Medicine, Zhejiang University from January 2023 to June 2024 as the study subjects. Based on the occurrence of reproductive tract infections, the patients were divided into a non-infection group and an infection group. Multivariate Logistic regression analysis and the decision tree method were employed to create a prediction model for GDM complicated with reproductive tract infections, and the risk prediction performance of the model was evaluated.Results:Among the 256 GDM patients, the age ranged from 20 to 43 (33.42 ± 4.25) years. The incidence of reproductive tract infections was 18.75% (48/256). Multivariate Logistic regression analysis revealed that age, glycosylated hemoglobin>6%, history of reproductive tract infections, uncontrolled blood glucose, and frequency of sexual activity ≥2 times during mid-to-late pregnancy were independent risk factors for reproductive tract infections in GDM patients ( χ2 values were 2.08-4.24, all P<0.05). The decision tree model for GDM complicated with reproductive tract infections consisted of 4 layers and 17 nodes, with the nodes selecting five indicators: age, frequency of sexual activity during mid-to-late pregnancy, blood glucose control status, sharing of vulvar cleaning tools, and history of reproductive tract infections. Among these, age was the most significant predictor of reproductive tract infections in GDM patients. The area under the curve (AUC) value of the decision tree model for GDM complicated with reproductive tract infections was 0.948, while the AUC value of the multivariate logistic regression model was 0.927. There was no statistically significant difference between the AUC values of the decision tree model and the multivariate logistic regression model ( P>0.05). Conclusions:The constructed decision tree model for GDM patients developing genital tract infections has high application value and can serve as a basis for clinical screening of potential GDM patients at risk of genital tract infections.

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.