OBJECTIVE To establish a method for simultaneous determination of ceftazidime， avibactam and aztreonam concentrations in human plasma.METHODS Stable-isotope-labeled compounds ceftazidime-D 5 ， avibactam- 13 C 5 and aztreonam- 2 H 6 were used as internal standards. Plasma samples were pretreated by protein precipitation with methanol， followed by nitrogen blowing. A ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） technique was applied to develop a quantitative analytical method for simultaneous measurement of ceftazidime， avibactam and aztreonam in human plasma. An ACQUITY UPLC BEH C 18 column was used as the chromatographic column. Gradient elution was performed with 0.01% formic acid-acetonitrile solution as the mobile phase at a flow rate of 0.3 mL/min. The column temperature was set at 40 ℃， and the injection volume was 5 μL. An electrospray ionization source was operated in both positive and negative ion modes for multiple reaction monitoring. Method validation was carried out according to the Guidelines for Bioanalytical Method Validation contained in volume Ⅳ of the 2020 edition of Chinese Pharmacopoeia .RESULTS The linear ranges of ceftazidime， avibactam and aztreonam were 0.04-200， 0.05-50，0.01-50 μg/mL， respectively， with all correlation coefficients no less than 0.996. The relative error （RE） for accuracy was no more than 12.25% （ n ＝5）， and the RSD for precision was no more than 13.49% （ n ＝5）. The RSD of internal-standard-normalized matrix factors was less than 15.00% （ n ＝6）. The RE in stability tests was lower than 15.00% （ n ＝3）.CONCLUSIONS This study successfully established an UPLC-MS/MS method for simultaneous determination of ceftazidime， avibactam and aztreonam concentrations in human plasma. The method possesses favorable accuracy， precision and stability， and can provide methodological support for therapeutic drug monitoring and individualized medication in patients treated with these three combined drugs.

Non-infectious arthritis (NIS) mainly includes osteoarthritis and inflammatory arthritis. With the aging of the population, chronic disease management of NIS patients in China will face great challenges, such as long disease duration, multi-system involvement, high demand for personalized diagnosis and treatment, lack of patient awareness, and difficulty in managing co-morbidities. To address these challenges, a systematic strategy for chronic disease management of NIS needs to be developed at the national level, which encompasses increasing popularization of science, improving the referral system, using artificial intelligence (AI) technology to assist in early diagnosis, and implementing interventions including optimized pharmacological/surgical treatments and multidisciplinary collaboration. In the future, with the deepening of the research on the application of AI technology and novel biologics, the management of chronic disease management of NIS will become more intelligent and personalized.

Background Work-related musculoskeletal disorders (WMSDs) are one of the major occupational health problems faced by bus drivers and should receive special attention. Objective To explore the associations of weekly working hours and sleep quality with neck and lower back WMSDs among bus drivers, as well as assess the potential mediating role of sleep quality. Methods From June to December 2022, we recruited bus drivers from 5 subsidiaries of the Shenzhen Bus Group by convenient sampling method. Demographic characteristics, lifestyles, and work-related features of the bus drivers were collected through a questionnaire survey. The Pittsburgh Sleep Quality Index (PSQI) scale and the Musculoskeletal Disorders Survey Questionnaire were used to assess sleep quality and WMSDs respectively. Logistic regression models were applied to analyze the associations of weekly working hours and sleep quality with WMSDs in neck and lower back. Furthermore, mediation analysis was performed to investigate the role of sleep quality in the associations between weekly work hours and neck and lower back WMSDs. Results A total of 1792 valid questionnaires were collected, with a response rate of 95.07%. Among these, 89.3% of the bus drivers worked more than 40 h per week, and the positive rate of poor sleep quality was 28.2%. The overall positive rate of WMSDs within one year before the survey was 69.12%. Among different body regions, the leading positive rates were identified in neck and lower back regions, at 55.92% and 53.68%, respectively. The logistic regression model showed that compared to the bus drivers with weekly work hours ≤40 h, those weekly worked (40~48] h (OR=1.43, 95%CI: 1.02, 2.01; OR=1.65, 95%CI: 1.17, 2.34), (48~56] h (OR=2.69, 95%CI: 1.89, 3.83; OR=2.30, 95%CI: 1.62, 3.29) and ＞56 h (OR=2.63, 95%CI: 1.86, 3.74; OR=3.23, 95%CI: 2.27, 4.62) had significantly increased risk of WMSDs in neck and lower back. Additionally, when compared to those with good sleep quality, bus drivers with poor sleep quality had higher risks of both neck and lower back WMSDs (OR=4.08, 95%CI: 3.20, 5.23; OR=4.15, 95%CI: 3.26, 5.30, respectively). Further mediation analysis indicated that sleep quality partially mediated the associations between weekly working hours and both neck and lower back WMSDs, with 36.15% and 33.68% as the respective proportion of mediation. Conclusion The positive rate of WMSDs is high among bus drivers, and the most common WMSDs occur in neck and lower back. Long working hours and poor sleep quality are significantly associated with increased risks of neck and lower back WMSDs. Sleep quality may mediate the associations of weekly working hours with neck and lower back WMSDs.

Objective:To investigate the application value of needle-knife accurate fistulo-tomy (NKAF) for difficult biliary cannulation during endoscopic retrograde cholangiopancreato-graphy (ERCP).Methods:The retrospective and descriptive study was conducted. The clinicopatho-logical data of 137 patients with difficult biliary cannulation during ERCP at Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine between January 2021 and December 2022 were collected. There were 51 males and 86 females, aged (69±13)years. All 137 patients received NKAF for cannulation during ERCP. Observation indicators: (1) surgical situations; (2) complications. Measurement data with normal distribution were represented as Mean± SD. Count data were repre-sented as absolute numbers. Results:(1) Surgical situations. Of 137 patients, 136 cases had succe-ssful cannulation, 1 case had failed cannulation with NKAF following unsuccessful double-guidewire technique. In the 136 successful cases, the endoscope was straightened in 42 cases, left in a long position in 37 cases, and maintained in the standard position in 57 cases. The cannulation time was (90±8)s. (2) Complications. The serum amylase at postoperative 6 hours in the 136 successful cases was (163±23)U/L. No patient developed post-ERCP pancreatitis. Of the 136 patients with successful cannulation, one case experienced post-sphincterotomy bleeding, which was observed oozing from the papillary orifice on emergency gastroscopy. The patient was successfully controlled with endoscopic clips.Conclusion:NKAF is safe and effective for difficult biliary cannulation during ERCP.

To analyze the epidemiological characteristics of COVID-19 in Fujian Province from the 49th week of 2022 to the 5th week of 2023,after further optimization of China's COVID-19 prevention and control measures on December 7,2022(the 49th week of 2022),this study used multi-dimensional surveillance data to dynamically assess population infection levels and their changing trends.The aim of the study was to provide a scientific basis for early warning of epidemic risk,medical resource allocation,and evalu-ation of socio-economic impact.A multi-source data surveillance system was constructed,encompassing surveillance of fever clinics at medical institutions(weekly collection of visits,positive nucleic acid and antigen test results,inpatients,and severe cases in sec-ondary or above hospitals),population nucleic acid test monitoring(weekly person-times and positivity rates of single-tube tests from the provincial system),sentinel hospital monitoring(weekly proportion of influenza-like illness visits at 18 sentinel hospitals and re-lated viral testing data),and monitoring of novel coronavirus variants(weekly systematic collection of genomic sequences of local and imported cases).Line charts were plotted weekly,and time series analysis,molecular epidemiological methods,and an improved SEIAR model were used to simulate epidemic spread.During the study period,the COVID-19 epidemic in Fujian Province exhibited three distinct stages.In the infection peak stage(52nd week of 2022),the provincial fever clinic visits reached 606 893 person-times,and a 49.2%positivity rate in population single-tube nucleic acid tests and 63.8%positivity rate in sentinel hospital monitoring were observed.In the medical load peak stage(2nd week of 2023),274 460 inpatients and 28 487 severe cases were recorded.In the epidemic decline stage(4th to 5th weeks of 2023),fever clinic visits decreased by 96.3%with respect to the peak,the single-tube nucleic acid test positivity rate decreased to 6.3%,and the sentinel hospital COVID-19 nucleic acid test positivity rate was 6.4%.All 508 sequenced local cases were Omicron variants,predominantly BA.5.2 and its sub-lineages(67.4%).Among 56 imported se-quenced cases,BA.5.2 and its sub-lineages accounted for 50.0%,and 16.1%comprised nine variants of interest including XBB and BQ.The model predicted the infection peak in the 52nd week of 2022,whereas the hospitalization peak lagged by approximately 10.6 days.Multi-source data monitoring revealed a three-stage development of the COVID-19 epidemic in Fujian.The BA.5.2 strain was dominant during the epidemic.The combination of multi-source monitoring data and modeling provides important references for epi-demic prevention and control,and highlights the need to improve the monitoring system in follow-up.

Objective To explore the effect of exercise under cold exposure on hepatic protein ki-nase B(AKT)/forkhead box O1(FoxO1)expression in obese rats.Methods Among rats successfully induced nutritional obesity by high-fat diet,forty were selected and randomly divided into a normal-temperature control group(NC,n=10),a normal-temperature exercise group(NE,n=10),a sus-tained-cold control group(SC,n=10),and a sustained-cold exercise group(SE,n=10).The normal temperature was kept at 25°C±1°C,while the low temperature remained at 4°C±1°C,with 50%to 60%relative humidity.The exercise protocol was every other day at a speed of 25 m/min for 2 sets of 30 min each,with an interval of 10 minutes.After 5 weeks,glucose and insulin tolerance were tested by oral glucose tolerance test(OGTT)and insulin tolerance test insulin tolerance test(ITT).Then,all rats were weighed and sacrificed,then taken blood from the abdominal aorta to sepa-rate serum,followed by detection of serum alanine aminotransferase(ALT)and aspartate aminotransfer-ase(AST)levels using fully automatic biochemical analyzer.Moreover,livers were weighed to calcu-late the liver index.Meanwhile,the mRNA expressions of hepatic AKT,FoxO1 and PEPCK were de-tected using RT-qPCR,while the protein expressions of AKT,phosphorylated protein kinase B(p-AKT),and FoxO1 in the liver were measured using Western blotting.Results(1)The average body weights of the NE,SC and SE groups were significantly lower than the NC group(P<0.01),with that of the SC and SE groups significantly lower than the NE group(P<0.01).(2)Compared with the NC group,the area under the OGTT curve of the SC group decreased(P<0.01).Moreover,the area under the ITT curve of the SE group was significantly lower than the other 3 groups(P<0.01,P<0.01,P<0.05),with that of the NE and SC groups significantly lower than the NC group(P<0.05,P<0.01).(3)The liver indices of the NE,SC and SE groups were all significantly lower than the NC group(P<0.01),while the serum ALT level of the NE group was significantly lower than the NC and SE groups(P<0.05),with that of the SC group significantly lower than the NC group(P<0.01).(4)Compared with the NC group,hepatic AKT mRNA expression increased significantly in the SE group(P<0.05),while the hepatic FoxO1 and PEPCK mRNA expression decreased significantly in the other three groups(P<0.01).(5)Compared with the NC group,the liver AKT protein phosphorylation levels were significantly higher in the other three groups(P<0.05,P<0.05,P<0.01),while the FoxO1 protein expression decreased significantly(P<0.05,P<0.01,P<0.01).(6)Body mass,liver FoxO1,PEPCK mRNA expression,AKT protein phosphorylation level and FoxO1 protein expression level of obese rats were affected by cold exposure,exercise and cold exposure+exercise,and the liver index and serum ALT level were done by exercise and cold exposure+exercise.However,the area un-der the ITT curve and liver AKT mRNA content were impacted by cold exposure and exercise,while that under the OGTT curve was influenced by cold exposure and cold exposure+exercise.Conclu-sion Exercise under cold exposure can activate hepatic AKT/FoxO1 signaling pathway,protect liver function,increase insulin sensitivity,and effectively improve glucose metabolism in obese rats.

The specific mechanisms of interferon regulatory factor 8(IRF8)in porcine intestinal in-nate immunity and resistance to enteric virus infection remain to be elucidated.To investigate the immunoregulatory role of IRF8,establishing an IRF8 gene knockout porcine intestinal epithelial cell(IPEC-J2)monoclonal cell line is of significant importance.This study initially aimed to obtain recombinant adeno-associated virus rAAV-sgIRF8-eGFP capable of knocking out the IRF8 gene through co-transfection of HEK-293T cells with three plasmids.Subsequently,IPEC-J2 cells were infected with the virus,and those expressing eGFP were selected by flow cytometry and cultured to form monoclonal cell lines.These cell lines were then identified by Sanger sequencing and West-ern blot techniques.Lastly,qPCR analysis was used to measure the expression levels of interferon factors IFN-α,IFN-β,IFN-γ and IFN-λ,providing preliminary insights into the impact of IRF8 gene knockout on IPEC-J2 cell immunity.The results demonstrated successful generation of rAAV-sgIRF8-eGFP,which successfully infected IPEC-J2 cells leading to eGFP fluorescence.Flow cytometry followed by cell culture led to the establishment of two monoclonal cell lines,IRF8-KO1 and IRF8-KO3.Sanger sequencing revealed a five-base deletion in IRF8-KO1 and a seven-base dele-tion in IRF8-KO3.Western blot confirmed the absence of IRF8 protein expression in IRF8-KO1,making it an ideal candidate monoclonal cell line.qPCR analysis of interferon factors indicated sig-nificant decrease in IFN-γ(P＜0.05)and IFN-λ(P＜0.01)transcription level in IRF8-knockout cells,while the transcription levels of IFN-α and IFN-β remained relatively unchanged.This study successfully established an IRF8 gene knockout IPEC-J2 monoclonal cell line,providing a founda-tion for further research on IRF8-related porcine intestinal immune regulation and mechanisms of intestinal virus infection.

This paper analyzed the clinical data of a patient with recurrent infection of chlamydia psittaci pneumonia within 7 months. The patient had a clear history of contact with live poultry, and the clinical manifestations were dry cough, persistent fever, and respiratory failure. Chest CT imaging changes showed lobar consolidation of infected lung lobes, ground-glass shadows, bronchial air-filling signs, and pleural effusion. The two infections were detected in bronchoalveolar lavage fluid by metagenomic next-generation sequencing (mNGS) and pathogen targeted next-generation sequencing (tNGS), respectively, to achieve early diagnosis of chlamydia psittaci pneumonia. New tetracycline drugs were used as the core of treatment for both infections, and rapid improvement was achieved after anti-infection treatment.

Objective To compare the effects of different doses of budesonide and formoterol fumarate powder for inhalation combined with montelukast sodium tablet in the treatment of cough variant asthma(CVA)and the improvement of airway function and inflammatory factors.Methods Elderly patients with cough variant asthma were randomly divided into group A and group B.Both groups of patients received budesonide and formoterol fumarate powder for inhalation combined with montelukast sodium tablet.Group A was given budesonide and formoterol fumarate powder for inhalation(Ⅱ),2 inhalation per time,twice a day;Group B was given budesonide and formoterol fumarate powder for inhalation,4 inhalation per time,twice a day;budesonide fumatrol inhalation powder mist for continuous treatment for 6 months,and montelukast sodium tablet 10 mg once a day for at least 3 months.The nighttime cough scores of the two groups were compared before treatment and after treatment.The percentage of forced expiratory volume in one second(FEV1)in the predicted value,the maximum mid expiratory flow(MMEF),the fractional exhaled nitric oxide(FeNO),interleukin-5(IL-5)and eosinophils were compared between the two groups.The incidence of adverse drug reactions and the recurrence rate within 1 year were compared between the two groups.Results A total of 45 cases were enrolled in both the group A and the group B.At 9 months after treatment,the nocturnal cough scores of the group A and the group B were(0.93±0.42)and(0.65±0.29)points,respectively;the percentage of FEV1 in the predicted value were(97.75±9.67)％and(100.93±11.06)％,respectively;the MMEF values were(2.81±1.04)and(3.08±1.09)L·s-1,respectively;the FeNO values were(18.94±9.75)and(15.94±7.96)ppb,respectively;the IL-5 levels were(10.88±7.06)and(8.11±5.56)pg·mL-1,respectively.The above indicators in group B showed statistically significant differences compared to group A(all P＜0.05).The total incidence of adverse drug reactions in group A and group B were 8.89％(5 cases/45 cases)and 13.33％(6 cases/45 cases),respectively.The recurrence rates was 15.56％(7 cases/45 cases)and 13.33％(6 cases/45 cases),respectively.There was no statistically significant difference in the above indicators between group B and group A(all P＞0.05).Conclusion For elderly patients with CVA,higher dose of budesonide and formoterol fumarate powder for inhalation combined with montelukast sodium tablet can better improve cough symptoms,reduce the level of airway hyperresponsiveness and inflammatory factors,reduce the recurrence rate,and the patients are well tolerated.

Objective:To explore the impacts of smoking on the need for respiratory support and mortality in patients hospitalized with pneumonia.Methods:A total of 24 367 patients hospitalized with pneumonia from 2009 to 2017 in the China Kadoorie Biobank, were included. Smoking status was self-reported, and data regarding respiratory support during hospitalization and mortality during follow-up were obtained from medical claims and death registries, respectively. OR, HR, and 95% CI were calculated and adjusted for potential confounders using logistic regression models and Cox proportional hazards regression models, respectively. Results:Among males, current smokers or those who quit smoking due to illness had higher risks of requiring respiratory support ( OR=1.15, 95% CI: 1.03-1.29), 1-year mortality ( HR=1.66, 95% CI: 1.32-2.08), and 5-year mortality ( HR=1.32, 95% CI: 1.13-1.54) following pneumonia hospitalization compared to nonsmokers. Male smokers who started smoking at a younger age or with longer smoking duration had the highest mortality risks (trend test both P<0.05). Female current smokers or those who quit smoking due to illness had higher risks of 1-year mortality ( HR=1.62, 95% CI: 1.17-2.23) and 5-year mortality ( HR=1.33, 95% CI: 1.06-1.67). We found no statistically significant difference in 90-day mortality between current smokers/those who quit smoking due to illness and nonsmokers. Conclusions:Smoking was associated with higher risks of requiring respiratory support and mortality in patients hospitalized with pneumonia, especially among males and heavy smokers. These findings highlight the need for targeted strategies to promote smoking cessation in patients hospitalized with pneumonia.