Objective To investigate the gene expression profile of cervical exfoliated cells from woman treated by artificial cycle,and their potential association with endometrial receptivity in order to screen specific biomarkers closely related to the receptivity.Methods A total of 19 female patients were enrolled from those preparing for frozen embryo transfer(FET)at the Reproductive Center of First Medical Center of Chinese PLA General Hospital from February 2024 to October 2024.Under the artificial cycle frozen embryo transfer protocol,the endometrial tissues were collected on the 4th day after progesterone administration(P+4)to verify their endometrial receptivity status.Additionally,cervical exfoliated cells were collected on the 4th day(P+4)and the 6th day(P+6)after progesterone administration.RNA sequencing(RNA-Seq)was used to detect gene expression profiles.Differentially expressed genes(DEGs)were identified using the criteria of|log2fold change|＞1 and a false discovery rate(FDR)＜0.05,followed by bioinformatics analysis.The protein-protein interaction(PPI)network of DEGs was constructed using R software(4.4.1)and analyzed with gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)analyses.The candidate genes were identified based on the PPI network using Cytoscape software.Quantitative reverse transcription PCR(RT-qPCR)was employed to validate the target candidate genes both in vitro and in vivo.Results The rsERT confirmed that all 19 women were in state of endometrial receptivity at P+6.RNA-Seq identified 3 458 DEGs in cervical exfoliated cells between P+4 and P+6.The up-regulated DEGs were mainly enriched in the pathways associated with immune response and cell differentiation,and the down-regulated ones were mainly enriched in the pathways associated with lipid metabolism and cell proliferation.Using maximal clique Centrality(MCC)algorithm in the PPI network,the top 20 genes were selected.Among them,6 genes,such as IFIT2,OASL,MX1,RSAD2,IFIT1 and IFIT3,tied for the first place,and the 6 genes all belong to interferon-stimulated genes(ISGs).qRT-PCR indicated that the above 6 genes showed significantly higher expression levels in the cervical exfoliated cells at the P+4 stage than the cells at the P+6 stage(P＜0.05).Conclusion There are changes in the expression levels of the genes related to immunity and cytoskeleton remodeling in cervical exfoliated cells during the endometrial receptivity phase.The decrease in the expression of ISGs may serve as a potential biomarker for endometrial receptivity.

Objective:To investigate the efficacy and potential mechanism of sulfasalazine (SASP) therapy for intrahepatic cholestasis.Methods:Forty SD rats were randomly divided into a normal group (carboxymethylcellulose sodium 0.5%), a model group (carboxymethylcellulose sodium 0.5%), a SASP group (sulfasalazine 150 mg/kg), and an ursodeoxycholic acid (UDCA 100 mg/kg) group, with ten rats in each group. The cholestatic liver injury model was induced using α-naphthylisothiocyanate. Blood samples were collected to detect liver biochemistry and cholestasis indexes. Rat liver tissue was collected for hematoxylin-eosin staining and Mason staining. Liver tissue was analyzed using transcriptome sequencing, real-time reverse transcription quantitative polymerase chain reaction, Western blotting and flow cytometry. Simultaneously, the level of inflammatory factors, total cholesterol, and total bile acids were measured in liver tissue. A t-test or a nonparametric test was selected based on the distribution and variance characteristics of the data. Results:The serum levels of alanine aminotransferase [(386.88±155.77) U/L], aspartate aminotransferase [(593.13±251.44) U/L], alkaline phosphatase [(561.25±167.54) U/L], total bilirubin [(38.00±29.75) mol/L] and total bile acids [(191.31±91.48) mol/L] were significantly lower in the SASP than the model groups [(778.75±313.59) U/L, (1 159.38±274.62) U/L, (801.25±161.28) U/L, (86.63±27.83) mol/L, (432.63±151.54) mol/L, P<0.05]. Liver histopathology showed that the inflammatory cells in the manifold area, the bile duct proliferation and dilation, and the collagen deposition in the manifold area were significantly improved under the pathological state of cholestasis in the SASP group. The results of transcriptome sequencing demonstrated that SASP activated the peroxisome proliferator actived receptor α (PPAR α) and inhibited Th17 cell differentiation. The PPARα mRNA level in the liver tissue of rats was significantly increased in the SASP group compared with that in the model group [(0.41±0.28) vs. (0.16±0.04), P<0.05], and the expression of 3-hydroxy-3-methylglutaryl coenzyme A reductase was decreased compared with that in the model group [(3.09±1.16) vs. (8.19±2.19), P<0.05], which was also verified at the protein level. The concentrations of total cholesterol [(0.31±0.34) mmol/g] and total bile acids [(2.58±0.99) μmol/g] were lower than the model group [(0.83±0.62) mmol/g and (4.07±0.91) μmol/g] ( P<0.05), and at the same time it was accompanied by lower levels of inflammatory factors ( P<0.05). SASP treatment decreased the expression of retinoic acid receptor-related orphan receptor γt gene ( P<0.05) and the proportion of Th17 ( P<0.05). Conclusion:SASP can improve cholestatic liver injury, and its mechanism is related to the activation of peroxisome proliferator-activated receptor α and the inhibition of Th17 cell differentiation.

Objective:To investigate the status of work-related musculoskeletal disorders (WMSDs) in male large-scale mechanical maintenance workers, and to explore the relationship between WMSDs and thyroid indexes.Methods:From April to July 2022, male front-line maintenance workers in a large-scale mechanical maintenance enterprise who participated in occupational health examination were selected as the study subjects ( n=2036). The occurrence of WMSDs was investigated by questionnaire. The levels of triiodothyronine (T3), thyroxine (T4) and thyroid stimulating hormone (TSH) in serum were detected. χ2 test, t-test and Mann-Whitney U test were used to detect thyroid diseases and hormone levels of workers in WMSDs group and non-WMSDs group, and binary logistic regression model was used to analyze the relationship between thyroid disease, T3, T4, TSH and WMSDs. Results:The incidence of WMSDs among the male large-scale mechanical maintenance workers was 73.18% (1490/2036). The rate of thyroid disease in WMSDs group was higher than that in non-WMSDs group [8.26% (123/1490) vs. 4.95% (27/546), χ 2=6.42, P=0.011], and the TSH value was lower than that in non-WMSDs group[1.66 (1.23, 2.26) μIU/ml vs. 1.75 (1.30, 2.42) μIU/ml, Z=-2.40, P=0.019]. There were no significant differences in the levels of T3, T4 and abnormal levels of 3 hormones between the two groups ( P>0.05). After accounting for individual and occupational factors, workers with thyroid disease had an increased risk of WMSDs ( OR=1.656, 95% CI: 1.072-2.559, P=0.023), while workers with increased TSH had a decreased risk of WMSDs ( OR=0.897, 95% CI: 0.823-0.977, P=0.013) . Conclusion:The incidence of WMSDs in male workers of large-scale mechanical maintenance is high, and its incidence may be related to thyroid index. Thyroid disease may be a risk factor for WMSDs, and increased TSH may be a protective factor for WMSDs.

Objective To investigate the efficacy of voriconazole in the treatment of pulmonary tuberculosis complicated with chronic pulmonary aspergillosis(CPA)based on CYP2C19 gene polymorphism detection and examine the factors affecting the efficacy for improving targeted therapy in clinical practice.Methods A total of 207 patients with pulmonary tuberculosis complicated with CPA treated in the Third People's Hospital of Kunming from December 2018 to November 2022 were randomly assigned to an observation group(105 cases)or a control group(102 cases).The patients in the control group received standard voriconazole treatment,while the patients in the observation group had their voriconazole regimen tailored based on CYP2C19 genotyping results.Plasma drug concentration levels,efficacy,and safety were compared between the two groups and in terms of CYP2C19 genotypes.Logistic regression analysis was used to identify the factors affecting treatment efficacy.Results The observation group showed significantly higher plasma voriconazole concentrations and overall antifungal efficacy compared to the control group(P＜0.05).In the observation group,CYP2C19 genotyping identified 37 extensive metabolizers(EM),47 intermediate metabolizers(IM),and 21 poor metabolizers(PM).Plasma concentration of voriconazole did not show significant difference between EM and IM(P＞0.05),but both PM and IM were associated with significantly lower plasma concentration of voriconazole than PM(P＜0.05).The clinical efficacy rate was 100％for PM,91.5％for IM,and 83.8％for EM(P＜0.05).The incidence of adverse events did not show significant difference among the three genotypes(P＞0.05).Logistic regression analysis revealed that lung cavitation,hypoalbuminemia,and agranulosis were significantly correlated with therapeutic efficacy(P＜0.05).Conclusions CYP2C19 gene polymorphism detection is valuable in clinical practice.It can inform anti-aspergillus therapy with voriconazole to effectively improve symptoms and clinical efficacy in patients with pulmonary tuberculosis complicated with CPA.Meanwhile,clinicians should be aware of the factors such as hypoproteinemia,agranulocytosis,and lung cavitation that may affect the efficacy of voriconazole.

Objective To investigate the efficacy of voriconazole in the treatment of pulmonary tuberculosis complicated with chronic pulmonary aspergillosis(CPA)based on CYP2C19 gene polymorphism detection and examine the factors affecting the efficacy for improving targeted therapy in clinical practice.Methods A total of 207 patients with pulmonary tuberculosis complicated with CPA treated in the Third People's Hospital of Kunming from December 2018 to November 2022 were randomly assigned to an observation group(105 cases)or a control group(102 cases).The patients in the control group received standard voriconazole treatment,while the patients in the observation group had their voriconazole regimen tailored based on CYP2C19 genotyping results.Plasma drug concentration levels,efficacy,and safety were compared between the two groups and in terms of CYP2C19 genotypes.Logistic regression analysis was used to identify the factors affecting treatment efficacy.Results The observation group showed significantly higher plasma voriconazole concentrations and overall antifungal efficacy compared to the control group(P＜0.05).In the observation group,CYP2C19 genotyping identified 37 extensive metabolizers(EM),47 intermediate metabolizers(IM),and 21 poor metabolizers(PM).Plasma concentration of voriconazole did not show significant difference between EM and IM(P＞0.05),but both PM and IM were associated with significantly lower plasma concentration of voriconazole than PM(P＜0.05).The clinical efficacy rate was 100％for PM,91.5％for IM,and 83.8％for EM(P＜0.05).The incidence of adverse events did not show significant difference among the three genotypes(P＞0.05).Logistic regression analysis revealed that lung cavitation,hypoalbuminemia,and agranulosis were significantly correlated with therapeutic efficacy(P＜0.05).Conclusions CYP2C19 gene polymorphism detection is valuable in clinical practice.It can inform anti-aspergillus therapy with voriconazole to effectively improve symptoms and clinical efficacy in patients with pulmonary tuberculosis complicated with CPA.Meanwhile,clinicians should be aware of the factors such as hypoproteinemia,agranulocytosis,and lung cavitation that may affect the efficacy of voriconazole.

Objective:To investigate the efficacy and potential mechanism of sulfasalazine (SASP) therapy for intrahepatic cholestasis.Methods:Forty SD rats were randomly divided into a normal group (carboxymethylcellulose sodium 0.5%), a model group (carboxymethylcellulose sodium 0.5%), a SASP group (sulfasalazine 150 mg/kg), and an ursodeoxycholic acid (UDCA 100 mg/kg) group, with ten rats in each group. The cholestatic liver injury model was induced using α-naphthylisothiocyanate. Blood samples were collected to detect liver biochemistry and cholestasis indexes. Rat liver tissue was collected for hematoxylin-eosin staining and Mason staining. Liver tissue was analyzed using transcriptome sequencing, real-time reverse transcription quantitative polymerase chain reaction, Western blotting and flow cytometry. Simultaneously, the level of inflammatory factors, total cholesterol, and total bile acids were measured in liver tissue. A t-test or a nonparametric test was selected based on the distribution and variance characteristics of the data. Results:The serum levels of alanine aminotransferase [(386.88±155.77) U/L], aspartate aminotransferase [(593.13±251.44) U/L], alkaline phosphatase [(561.25±167.54) U/L], total bilirubin [(38.00±29.75) mol/L] and total bile acids [(191.31±91.48) mol/L] were significantly lower in the SASP than the model groups [(778.75±313.59) U/L, (1 159.38±274.62) U/L, (801.25±161.28) U/L, (86.63±27.83) mol/L, (432.63±151.54) mol/L, P<0.05]. Liver histopathology showed that the inflammatory cells in the manifold area, the bile duct proliferation and dilation, and the collagen deposition in the manifold area were significantly improved under the pathological state of cholestasis in the SASP group. The results of transcriptome sequencing demonstrated that SASP activated the peroxisome proliferator actived receptor α (PPAR α) and inhibited Th17 cell differentiation. The PPARα mRNA level in the liver tissue of rats was significantly increased in the SASP group compared with that in the model group [(0.41±0.28) vs. (0.16±0.04), P<0.05], and the expression of 3-hydroxy-3-methylglutaryl coenzyme A reductase was decreased compared with that in the model group [(3.09±1.16) vs. (8.19±2.19), P<0.05], which was also verified at the protein level. The concentrations of total cholesterol [(0.31±0.34) mmol/g] and total bile acids [(2.58±0.99) μmol/g] were lower than the model group [(0.83±0.62) mmol/g and (4.07±0.91) μmol/g] ( P<0.05), and at the same time it was accompanied by lower levels of inflammatory factors ( P<0.05). SASP treatment decreased the expression of retinoic acid receptor-related orphan receptor γt gene ( P<0.05) and the proportion of Th17 ( P<0.05). Conclusion:SASP can improve cholestatic liver injury, and its mechanism is related to the activation of peroxisome proliferator-activated receptor α and the inhibition of Th17 cell differentiation.

Objective:To investigate the status of work-related musculoskeletal disorders (WMSDs) in male large-scale mechanical maintenance workers, and to explore the relationship between WMSDs and thyroid indexes.Methods:From April to July 2022, male front-line maintenance workers in a large-scale mechanical maintenance enterprise who participated in occupational health examination were selected as the study subjects ( n=2036). The occurrence of WMSDs was investigated by questionnaire. The levels of triiodothyronine (T3), thyroxine (T4) and thyroid stimulating hormone (TSH) in serum were detected. χ2 test, t-test and Mann-Whitney U test were used to detect thyroid diseases and hormone levels of workers in WMSDs group and non-WMSDs group, and binary logistic regression model was used to analyze the relationship between thyroid disease, T3, T4, TSH and WMSDs. Results:The incidence of WMSDs among the male large-scale mechanical maintenance workers was 73.18% (1490/2036). The rate of thyroid disease in WMSDs group was higher than that in non-WMSDs group [8.26% (123/1490) vs. 4.95% (27/546), χ 2=6.42, P=0.011], and the TSH value was lower than that in non-WMSDs group[1.66 (1.23, 2.26) μIU/ml vs. 1.75 (1.30, 2.42) μIU/ml, Z=-2.40, P=0.019]. There were no significant differences in the levels of T3, T4 and abnormal levels of 3 hormones between the two groups ( P>0.05). After accounting for individual and occupational factors, workers with thyroid disease had an increased risk of WMSDs ( OR=1.656, 95% CI: 1.072-2.559, P=0.023), while workers with increased TSH had a decreased risk of WMSDs ( OR=0.897, 95% CI: 0.823-0.977, P=0.013) . Conclusion:The incidence of WMSDs in male workers of large-scale mechanical maintenance is high, and its incidence may be related to thyroid index. Thyroid disease may be a risk factor for WMSDs, and increased TSH may be a protective factor for WMSDs.

Objective:To analyze the clinical features and genetic basis of a child with Disorder of sex development (DSD).Methods:A child who was admitted to the Linyi People′s Hospital for primary amenorrhoea on July 29, 2019 was selected as the study subject. Clinical data of the child was collected. Chromosomal karyotyping and quantitative real-time PCR were used to detect Y chromosome microdeletions and other chromosomal aberrations. Next-generation sequencing was carried out for the child and her parents. Candidate variant was verified by Sanger sequencing and bioinformatic analysis.Results:The child, a 13-year-old girl, has featured primary amenorrhoea and onset of secondary sex characteristics of males. Ultrasound exam had detected no uterus and definite ovarian structure, but narrow band vaginal hypoecho and curved cavernoid structure. The child was found to have a 46, XY karyotype without an AZF deletion. DNA sequencing revealed that she has harbored a maternally derived c. 323delA (p.Q108Rfs*188) variant in the nuclear receptor subfamily 5 group A member 1 ( NR5A1) gene, which may result in a truncated protein. The variant was classified as pathogenic (PVS1+ PM2_Supporting+ PP4) based on the guidelines from the American College of Medical Genetics and Genomics. Conclusion:The NR5A1: c. 323delA variant probably underlay the pathogenesis of 46, XY DSD in this child. The discovery of the novel variant has enriched the mutational spectrum of NR5A1 gene and provided a basis for clinical diagnosis, treatment and prenatal diagnosis.

Objective To investigate the pathogenic bacteria distribution,clinical features and risk factors of urinary tract infection in patients with ischemic stroke. Methods A retrospective study was conducted on 634 patients with ischemic stroke in Beijing Bo'ai Hospital from Janu-ary,2020 to December,2023.They were divided into control group(n=551,without urinary tract infection)and observation group(n=83,with urinary tract infection)according to whether they developed urinary tract infec-tion.The incidence of urinary tract infection,the distribution of pathogenic bacteria and the resistance of main pathogenic bacteria to different antibacterial drugs were analyzed.The clinical characteristics of the two groups were compared and analyzed.The independent risk factors of urinary tract infection in patients with ischemic stroke were analyzed with multivariate Logistic regression. Results A total of 83 cases of 634 patients with ischemic stroke developed urinary tract infection,and incidence was 13.09%.A total of 127 strains of pathogenic bacteria were isolated from the urine samples of the observation group,of which Gram-negative bacteria accounted for 62.99%(80/127),Gram-positive bacteria accounted for 20.47%(26/127)and strains of fungi accounted for 16.54%(21/127).The main Gram-negative pathogens were Escherichia coli and Klebsiella pneumoniae,which were high resistant to second-generation and third-generation cephalosporins,co-trimoxazole,and levofloxacin;moderately resistant to carbapenems,β-lactamase inhibitor compound preparation and aminoglycosides,etc.;and highly sensitive to tigecycline and polymyxin,etc.The main Gram-positive pathogens were Enterococcus faecalis and Enterococcus faecalis,which were a high resistant to erythromycin and gentamicin,and highly sensitive to linezolid,daptomycin,teicoplanin and vancomycin.The pathogenic fungi detected were not obviously resistant to common antifungal drugs.The proportion of female,di-abetes,indwelling catheter and neurogenic bladder were significantly higher in the observation group than in the control group(χ2>5.043,P<0.05).The female,diabetes,indwelling catheter and neurogenic bladder were inde-pendent risk factors for urinary tract infection in patients with ischemic stroke(P<0.05). Conclusion The pathogenic bacteria of patients with ischemic stroke with urinary tract infection are mainly Gram-nega-tive bacteria,followed by Gram-positive bacteria and fungi.The Gram-negative bacteria showed multiple drug re-sistance.Meanwhile,female,diabetes,indentured catheter and neurogenic bladder are the independent risk fac-tors for urinary tract infection.

Objective:To analyze the epidemiological and etiological characteristics of a death case of meningococcal meningitis in Hengyang city, Hunan Province in 2024.Methods:Epidemiological investigation of the death case was performed, and samples from the patient and close contacts were collected. Following cultivation and isolation, Neisseria meningitidis ( Nm) strains were analyzed by antimicrobial susceptibility testing, pulsed field gel electrophoresis (PFGE), and whole-genome sequencing for analyzing epidemiological and etiological characteristics. Phylogenetic analysis was carried out using core genomic multilocus sequence typing (cgMLST). Results:The case was a 16-year-old high school boarding student with fulminant meningococcal meningitis. He had shock symptoms, and died within 24 h of the onset of symptoms. Six Nm strains were isolated from the patient and his roommates, belonging to two distinct clades. Isolate 144569 from the patient was highly homologous to isolate 144572 from a close contact, both belonging to the highly pathogenic sublineage L44.1 of CC4821. The typical molecular features was C: P1.7-2, 14: F5-101: ST4821 (CC4821). The two strains carried the antimicrobial resistance genes of gyrA-71 and penA-552, indicating reduced susceptibility to quinolone and penicillin, which was with their resistance phenotype. The isolates from four close contacts clustered within the same clade, characterized by the molecular features of B: P1.18-25, 9-18: ST5829 (UA). Conclusions:The death case is caused by Nm serogroup C from highly pathogenic sublineage L44.1 of CC4821. The spread of this isolate has the potential risk of outbreaks of invasive meningococcal disease. It is necessary to enhanced the molecular epidemiological surveillance, particularly focusing on the transmission of multiple serogroups of Nm among adolescents and the increasing exposure risk.