Objective To establish a reversed-phase HPLC method for determining three stereoisomers in baloxavir marboxil and provide a basis for the quality specification of baloxavir marboxil.Methods The chromatographic column was CHIRALPAK IC-3(4.6 mm × 150 mm,3 μm),The mobile phase was acetonitrile-0.1％formic acid aqueous solution-isopropanol(35:50:15).The column temperature was 40 ℃.The flow rate was 0.5 mL·min-1.The injection volume was 10 μL.The detection wavelength was 259 nm.Result The isomer peaks were completely separated from the principal component peak.The detection limits for stereoisomers 1,2 and 3 were 0.024 7,0.038 7,0.038 1 μg·mL-1 respectively.The quantitation limits for stereoisomers 1,2 and 3 were 0.049 4,0.077 3,0.076 1 μg·mL-1 respectively.There were good linear relationships between the concentrations and peak area within the ranges of the study,and the linearity concentration ranges of stereoisomers 1,2 and 3 were 0.049 5-0.989 0 μg·mL-1,0.051 6-1.031 0 μg·mL-1,0.050 8-1.015 0 μg·mL-1 respectively.The linear correlation coefficients were 0.999 4.The recovery was 92.28％-103.90％.The sample solution was stable in 48 h at room temperature.Conclusion The method is accurate and reliable for determining stereoisomers in baloxavir marboxil,and provide a guideline of quality standards of baloxavir marboxil and safety evalution.

Aim To explore the potential mechanism of metformin on ATP-binding cassette transport A1(ABCA1)expression.Methods J774A.1 macrophages were treated with metformin and cycloheximide,and ABCA1 expression was determined by Western blot.His-tagged ABCA1 and HA-tagged Ub plasmids were co-transferred into HEK293 cells and stimulated with metformin.Co-immunoprecipitation(Co-IP)was used to test the binding ability of ABCA1 and ubiquitin.Candidate E3 ubiquitin-protein ligases(CE3)of ABCA1 were identified through Co-IP-based pro-teomics.The MIB1 plasmid was constructed and transferred into HEK293 cells,and Western blot was used to determine the effect of metformin and MIB1 on ABCA1 expression.Results Metformin increased the expression of ABCA1 in J774A.1 cells(P<0.01),and inhibited ABCA1 degradation(P<0.05).Metformin disrupted the binding of ABCA1 to ubiquitin(P<0.05).The proteins regulated by metformin in ABCA1 expression were primarily enriched in pathways re-lated to cell development,inflammation and immune defense.Metformin may upregulate ABCA1 protein expression via MIB1(P<0.05).Conclusion Metformin inhibits the degradation of ABCA1 by blocking the ubiquitin-proteasome system(UPS),and MIB1 might act as a candidate E3 ubiquitin-protein ligase(CE3)for ABCA1.

Objective:To observe the pregnancy outcomes of patients with unexplained recurrent spontaneous abortion (URSA) after interventional treatment or expectant treatment.Methods:This prospective study followed up 398 patients with recurrent spontaneous abortion from March 2017 to September 2022 in seven hospitals. Among them, 267 patients were diagnosed with URSA, including 124 patients who were initially diagnosed in the interventional treatment hospital and 143 patients who were initially diagnosed in the expectant treatment hospital. All URSA patients were followed up for 33 months. Ongoing pregnancy rates were observed as main outcome indicators.Results:A total of 127 patients became pregnant, and 107 of them had sustained pregnancies, the ongoing pregnancy rate was 84.25% (107/127). The ongoing pregnancy rate was 86.11% (31/36) in the interventional treatment group and 83.52% (76/91) in the expectant treatment group, with no significant difference ( P>0.05). During the follow-up, the ongoing pregnancy rates in the interventional treatment hospital and the expectant treatment hospital were 75.71% (53/70) and 94.74% (54/57), respectively, with a significant difference ( P<0.05). The ongoing pregnancy rate after interventional treatment in the interventional treatment hospital was 82.76% (24/29), which was similar to the 94.00% (47/50) after expectant treatment in the expectant treatment hospital ( P>0.05). Conclusion:The ongoing pregnancy rate of interventional treatment for URSA patients has not been significantly improved, suggesting that it may not be necessary to carry out this treatment.

Aim To explore the potential mechanism of metformin on ATP-binding cassette transport A1(ABCA1)expression.Methods J774A.1 macrophages were treated with metformin and cycloheximide,and ABCA1 expression was determined by Western blot.His-tagged ABCA1 and HA-tagged Ub plasmids were co-transferred into HEK293 cells and stimulated with metformin.Co-immunoprecipitation(Co-IP)was used to test the binding ability of ABCA1 and ubiquitin.Candidate E3 ubiquitin-protein ligases(CE3)of ABCA1 were identified through Co-IP-based pro-teomics.The MIB1 plasmid was constructed and transferred into HEK293 cells,and Western blot was used to determine the effect of metformin and MIB1 on ABCA1 expression.Results Metformin increased the expression of ABCA1 in J774A.1 cells(P<0.01),and inhibited ABCA1 degradation(P<0.05).Metformin disrupted the binding of ABCA1 to ubiquitin(P<0.05).The proteins regulated by metformin in ABCA1 expression were primarily enriched in pathways re-lated to cell development,inflammation and immune defense.Metformin may upregulate ABCA1 protein expression via MIB1(P<0.05).Conclusion Metformin inhibits the degradation of ABCA1 by blocking the ubiquitin-proteasome system(UPS),and MIB1 might act as a candidate E3 ubiquitin-protein ligase(CE3)for ABCA1.

Objective:To observe the pregnancy outcomes of patients with unexplained recurrent spontaneous abortion (URSA) after interventional treatment or expectant treatment.Methods:This prospective study followed up 398 patients with recurrent spontaneous abortion from March 2017 to September 2022 in seven hospitals. Among them, 267 patients were diagnosed with URSA, including 124 patients who were initially diagnosed in the interventional treatment hospital and 143 patients who were initially diagnosed in the expectant treatment hospital. All URSA patients were followed up for 33 months. Ongoing pregnancy rates were observed as main outcome indicators.Results:A total of 127 patients became pregnant, and 107 of them had sustained pregnancies, the ongoing pregnancy rate was 84.25% (107/127). The ongoing pregnancy rate was 86.11% (31/36) in the interventional treatment group and 83.52% (76/91) in the expectant treatment group, with no significant difference ( P>0.05). During the follow-up, the ongoing pregnancy rates in the interventional treatment hospital and the expectant treatment hospital were 75.71% (53/70) and 94.74% (54/57), respectively, with a significant difference ( P<0.05). The ongoing pregnancy rate after interventional treatment in the interventional treatment hospital was 82.76% (24/29), which was similar to the 94.00% (47/50) after expectant treatment in the expectant treatment hospital ( P>0.05). Conclusion:The ongoing pregnancy rate of interventional treatment for URSA patients has not been significantly improved, suggesting that it may not be necessary to carry out this treatment.

Objective To establish a reversed-phase HPLC method for determining three stereoisomers in baloxavir marboxil and provide a basis for the quality specification of baloxavir marboxil.Methods The chromatographic column was CHIRALPAK IC-3(4.6 mm × 150 mm,3 μm),The mobile phase was acetonitrile-0.1％formic acid aqueous solution-isopropanol(35:50:15).The column temperature was 40 ℃.The flow rate was 0.5 mL·min-1.The injection volume was 10 μL.The detection wavelength was 259 nm.Result The isomer peaks were completely separated from the principal component peak.The detection limits for stereoisomers 1,2 and 3 were 0.024 7,0.038 7,0.038 1 μg·mL-1 respectively.The quantitation limits for stereoisomers 1,2 and 3 were 0.049 4,0.077 3,0.076 1 μg·mL-1 respectively.There were good linear relationships between the concentrations and peak area within the ranges of the study,and the linearity concentration ranges of stereoisomers 1,2 and 3 were 0.049 5-0.989 0 μg·mL-1,0.051 6-1.031 0 μg·mL-1,0.050 8-1.015 0 μg·mL-1 respectively.The linear correlation coefficients were 0.999 4.The recovery was 92.28％-103.90％.The sample solution was stable in 48 h at room temperature.Conclusion The method is accurate and reliable for determining stereoisomers in baloxavir marboxil,and provide a guideline of quality standards of baloxavir marboxil and safety evalution.

OBJECTIVES: Several countries have successfully eliminated measles, and China is making significant strides toward achieving this goal. This study focused on investigating the patterns of measles infections in Zhejiang Province, China, as well as control measures. The objective was to provide valuable insights that could contribute to the development of nationwide elimination strategies. METHODS: We analyzed measles surveillance data from 2005 to 2022 in Zhejiang Province. We utilized a joinpoint regression model to examine trends in measles. Additionally, we employed SaTScan version 9.5 to identify spatial-temporal clusters. Finally, we used an age-period-cohort model to assess the effects of age, period, and cohort. RESULTS: The age-standardized incidence rate (ASIR) of measles infection in Zhejiang Province from 2005 to 2022 was 5.24 per 100,000, showing a consistent and significant downward trend with an annual percentage change of -24.93% (p<0.05). After 2020, the ASIR for measles infection fell to below 0.1 per 100,000. The majority of measles cases occurred in individuals either without an immunization history or with an unknown immunization status, representing 41.06% and 41.40% of the cases from 2010 to 2022, respectively. According to data from the National Measles Surveillance System, the annual rate of discarded measles cases from 2009 to 2014, and the annual rate of discarded measles and rubella cases from 2015 to 2022, were both above 2 per 100,000, indicating the high sensitivity of the measles surveillance system. CONCLUSIONS The significant reduction in measles incidence from 2005 to 2022 demonstrates substantial progress in Zhejiang Province towards the elimination of measles.

Dimethyl oxalate is one of the occupational toxic chemicals and causes strong renal toxicity. On May 16, 2023, a patient with acute dimethyl oxalate poisoning was admitted to Dingxi People's Hospital. The patient presented with nausea, vomiting, lumbar distension, weakness, poor appetite, anuria, and rapidly progressing acute kidney injury. Renal biopsy confirmed acute oxalate nephropathy. After symptomatic supportive treatments such as blood purification, anti-oxidative stress, glucocorticoid, fluid supplementation, alkalized urine, anti-infection, controlling blood pressure, calcium supplementation and anemia correction, the patient's symptoms disappeared, and the kidney function basically returned to normal. This case suggested that the etiology of patients with acute kidney injury must be clearly identified, and renal biopsy was an important examination method. For patients suffering from acute dimethyl oxalate poisoning, comprehensive treatment based on blood purification should be performed as soon as possible, aiming to improve the prognosis.

Background and objective Lung squamous cell carcinoma(LUSC)is a subtypes of non-small cell lung cancer(NSCLC).It has been reported that members of the protocadherin γ family can regulate tumor cell growth by inhibiting the Wnt signaling pathway.Protocadherin-gamma subfamily B4(PCDHGB4)as a family member in LUSC was rarely reported.The aim of this study was to investigate the role and potential prognostic value of PCDHGB4 in the development of LUSC us-ing bioinformatics methods.Methods The Cancer Genome Atlas(TCGA),cBioPortal and UALCAN databases were used to analyze the expression,prognosis,clinicopathological features,immune cell infiltration,immune regulatory genes,immune checkpoint inhibitors(ICIs),and methyltransferases of PCDHGB4 in LUSC.At the single cell level,we analyzed the clustering results of cell subtypes and the expression of PCDHGB4 in different immune cell subpopulations.In addition,we compared the promoter methylation levels of PCDHGB4 in LUSC tissues and normal tissues and performed protein-protein interaction and mutation analysis.Finally,enrichment analysis was performed based on the differentially expressed genes.Results Bioinformat-ics analysis results showed that the expression level of PCDHGB4 in LUSC tissues was lower than that in normal tissues.Sur-vival analysis showed that increased PCDHGB4 expression was associated with poor prognosis.Single-cell sequencing analysis showed that PCDHGB4 was expressed in T cells,monocytes or macrophages,and dendritic cells.It was further found that PCD-HGB4 played an important role in tumor immunity and confirmed that PCDHGB4 was associated with immune checkpoints,immune regulatory genes,and methyltransferases.Besides,enrichment analysis revealed that PCDHGB4 was involved in mul-tiple cancer-related pathways.Conclusion The expression of PCDHGB4 was low in LUSC.PCDHGB4 was related to the poor prognosis of patients,and PCDHGB4 was closely related to the infiltration and pathway of tumor immune cells.PCDHGB4 may be a potential prognostic marker and a new target for immunotherapy in LUSC.

Dimethyl oxalate is one of the occupational toxic chemicals and causes strong renal toxicity. On May 16, 2023, a patient with acute dimethyl oxalate poisoning was admitted to Dingxi People's Hospital. The patient presented with nausea, vomiting, lumbar distension, weakness, poor appetite, anuria, and rapidly progressing acute kidney injury. Renal biopsy confirmed acute oxalate nephropathy. After symptomatic supportive treatments such as blood purification, anti-oxidative stress, glucocorticoid, fluid supplementation, alkalized urine, anti-infection, controlling blood pressure, calcium supplementation and anemia correction, the patient's symptoms disappeared, and the kidney function basically returned to normal. This case suggested that the etiology of patients with acute kidney injury must be clearly identified, and renal biopsy was an important examination method. For patients suffering from acute dimethyl oxalate poisoning, comprehensive treatment based on blood purification should be performed as soon as possible, aiming to improve the prognosis.