AIM: To evaluate the diagnostic value of serum Th1-type cytokine levels and extraocular muscle-related parameters in thyroid-associated ophthalmopathy(TAO).METHODS: A retrospective study was conducted on 45 patients diagnosed with TAO in our hospital from January 2023 to December 2024, and 20 normal volunteers during the same period as controls. Venous blood samples of the patients were collected to detect the concentrations of Th1-type cytokines [interferon-γ(IFN-γ), tumor necrosis factor-α(TNF-α), interleukin-2(IL-2), and interleukin-12(IL-12)] in the serum. Additionally, the end diastolic velocity(Ved), velocity maximum(Vmax), resistance index(RI)of the central retinal artery, as well as the thickness and left-right diameter of the medial rectus muscle were measured. Logistic regression model was used to analyze the risk factors of TAO, and receiver operating characteristic curve(ROC)was adopted to evaluate the diagnostic efficacy of each index for the occurrence of TAO.RESULTS: The general information of the two groups was comparable. Compared with the normal control group, the serum concentrations of IFN-γ and TNF-α in TAO patients were significantly increased, Ved and Vmax were lower than those in the control group, and RI and the thickness of the medial rectus muscle were higher than those in the control group(all P<0.05). Logistic regression analysis showed that serum IFN-γ concentration, Ved, Vmax, and the thickness of the medial rectus muscle were all risk factors for TAO. ROC curve analysis indicated that the AUCs of serum IFN-γ concentration, Ved, Vmax, and the thickness of the medial rectus muscle for the diagnostic efficacy of TAO were 0.756, 0.769, 0.732, and 0.642, respectively. The combined detection of IFN-γ, Ved, Vmax, and the thickness of the medial rectus muscle had an AUC of 0.840 and a Youden index of 0.59, which was superior to the detection of a single indicator.CONCLUSION: The levels of serum Th1-type cytokines and extraocular muscle-related ultrasound indicators have certain value in the diagnosis of TAO. The combination of IFN-γ, Ved, Vmax, and the thickness of the medial rectus muscle has better diagnosis efficiency in TAO, which can provide a certain reference for the early diagnosis of TAO.

Objective To establish a sandwich enzyme-linked immunosorbent assay (ELISA) method for the quantitative detection of Chromogranin A (CHGA) in saliva, and to explore its preliminary application in the testing of saliva samples. Methods Recombinant human CHGA protein was used to immunize BALB/c mice, and monoclonal antibodies (mAbs) were prepared and screened using conventional hybridoma technology. A double-antibody sandwich ELISA detection method was constructed, and the matrix effect of saliva samples was optimized. This method was then applied to detect the concentration of CHGA in the saliva of stressed individuals. Results Twenty-one stable hybridoma cell lines secreting high affinity anti-human CHGA antibodies were obtained. A pair of detection antibodies with the best effect was selected, and the optimal coating concentration was determined to be 10 μg/mL, with the optimal dilution of detection antibodies being 1:32 000. The accuracy and reproducibility of this method were verified, with both intra-batch and inter-batch variation coefficients less than 15×, and the recovery rate between 80× and 120×. The matrix effect was further optimized to make it suitable for saliva sample detection. Saliva samples from individuals in different stress states were collected, and the CHGA levels were detected using the method established in this study, indicating its potential to reflect the intensity of stress. Conclusion A reliable saliva CHGA ELISA detection method has been successfully established, and its potential as a biomarker in stress-related research has been preliminarily explored.
Saliva/metabolism* ; Enzyme-Linked Immunosorbent Assay/methods* ; Humans ; Animals ; Mice, Inbred BALB C ; Mice ; Chromogranin A/immunology* ; Antibodies, Monoclonal/immunology* ; Female ; Male ; Reproducibility of Results ; Adult

The regulatory processes in developmental biology research are significantly influenced by long non-coding RNAs (lncRNAs). However, the dynamics of lncRNA expression during human tooth development remain poorly understood. In this research, we examined the lncRNAs present in the dental epithelium (DE) and dental mesenchyme (DM) at the late bud, cap, and early bell stages of human fetal tooth development through bulk RNA sequencing. Developmental regulators co-expressed with neighboring lncRNAs were significantly enriched in odontogenesis. Specific lncRNAs expressed in the DE and DM, such as PANCR, MIR205HG, DLX6-AS1, and DNM3OS, were identified through a combination of bulk RNA sequencing and single-cell analysis. Further subcluster analysis revealed lncRNAs specifically expressed in important regions of the tooth germ, such as the inner enamel epithelium and coronal dental papilla (CDP). Functionally, we demonstrated that CDP-specific DLX6-AS1 enhanced odontoblastic differentiation in human tooth germ mesenchymal cells and dental pulp stem cells. These findings suggest that lncRNAs could serve as valuable cell markers for tooth development and potential therapeutic targets for tooth regeneration.
Humans ; RNA, Long Noncoding/metabolism* ; Odontogenesis/genetics* ; Tooth Germ/embryology* ; Cell Differentiation ; Gene Expression Regulation, Developmental ; Mesoderm/metabolism* ; Tooth/embryology* ; Gene Expression Profiling ; Sequence Analysis, RNA ; Dental Pulp/cytology*

OBJECTIVE To get the current situation of healthcare associated infection(HAI)and implement targe-ted management through analyzing the trends of HAI incidence rate,the distribution of clinical departments,the sites of HAI and the composition of pathogens in a children's hospital in a five-year period.METHODS Data on HAI cases were collected from all inpatients in Children's Hospital of Soochow University from 2019 to 2023 through the real-time monitoring system of XINGLIN Healthcare acquired Infection Surveillance;Cochran-Armit-age test was used to check for the significant changes of distribution of HAI cases.RESULTS A total of 383,376 hospitalized patients were monitored from 2019 to 2023,of which HAI occurred on 6670 cases and 7209 case-times with an incidence rate of 1.74%and an incidence case rate of 1.88%.The top five departments of HAI inci-dence rates were cardiac,neonatal,surgical and pediatric intensive care unit and department of hematology.HAI mainly occurred in blood and alimentary system,upper and lower respiratory tract.A total of 2668 strains of path-ogenic bacteria were isolated,of which 1346 strains were gram-negative bacteria,1140 strains were gram-positive bacteria and 182 strains were fungi.The top three gram-negative bacteria were Klebsiella pneumoniae,Escherich-ia coli and Pseudomonas aeruginosa;the top three gram-negative pathogens were Streptococcus,Staphylococcus aureus and Staphylococcus epidermidis;and the top three fungi were Candida albicans,Candida parapsilosis and Candida tropicalis.CONCLUSIONS The HAI incidence rate of this hospital steadily declines in the past five years,however the same changes are not observed in the departments with high incidence of HAI.Attention should be paid to the raising bloodstream infections and detection rate of S.epidermidis.

Objective:To investigate the clinical distribution characteristics changes in antimicrobial resistance, and carbapenemase resistance genes of Klebsiella pneumoniae isolated from children in Chongqing region during the period of January 2019 to December 2024, providing a basis for the rational use of antibiotics and the prevention and control of nosocomial infections.Methods:An observational study was conducted to retrospectively analyze 5 020 Klebsiella pneumoniae (KP) isolates detected in four hospitals of the Southwest Pediatric Laboratory Specialty Alliance. Antimicrobial susceptibility testing was performed by the minimum inhibitory concentration method combined with the disk diffusion method. Results were interpreted according to the 2024 Clinical and Laboratory Standards Institute (CLSI) standards. Carbapenemase resistance genes were detected by polymerase chain reaction (PCR) combined with Sanger sequencing. WHONET 5.6 was used for resistance analysis and SPSS 19.0 for statistical analysis. The chi-square test was used to assess trends in resistance rates, ESBL detection rates, and resistance rates of different CRKP carbapenemase genotypes from 2019 to 2024. Statistical significance was confirmed if the two-tailed P-value was <0.05. Results:A total of 5 020 strains were isolated, with a detection rate of 5.1% (5 020/99 063). The majority were from sputum (59.2%, 2 970/5 020), followed by pus (17.1%, 857), urine (9.7%, 487), venous blood (6.5%, 326), secretions (2.6%, 130), and other specimens (5.0%, 250).The lowest resistance rate was to amikacin (3.8%), followed by levofloxacin (10.9%), imipenem (19.1%), and meropenem (19.9%). Resistance rates to cefoperazone/sulbactam ( χ2=9.982 0, P=0.001 6), piperacillin/tazobactam ( χ2=10.110 0, P=0.001 5), ceftazidime ( χ2=3.849 0, P=0.049 8), cefotaxime ( χ2=7.605 0, P=0.005 8), cefepime ( χ2=13.510 0, P=0.000 2), aztreonam ( χ2=6.457 0, P=0.011 1), imipenem ( χ2=4.672 0, P=0.030 7), and levofloxacin ( χ2=7.555 0, P=0.006 0) showed an annual increasing trend. The main carbapenemase genes were blaNDM-5 (42.2%, 127/301), blaNDM-1 (33.9%, 102/301), and blaKPC-2 (17.3%, 52/301). Patients with KPC-2-producing strains (median age, 240 days) were older than those with NDM-1/NDM-5-producing strains (median age, 40 days) ( χ2=22.620 0, P<0.000 1). In neonatal wards, the detection rate of NDM-KP was higher than that of KPC-KP (64.6%, 148/229 vs. 26.9%, 14/52, χ2=24.680 0, P<0.000 1), whereas in ICUs, it was lower (6.1%, 14/229 vs. 26.9%, 14/52, χ2=20.450 0, P<0.000 1). Conclusion:In Chongqing region, the isolation rate of K. pneumoniae from sputum was the highest with most cases from neonatal wards. Resistance to carbapenems showed an upward trend. BlaNDM-5 was the predominant genotype in pediatric CRKP. Patients with KPC-KP were older than those with NDM-KP. NDM-KP predominated in neonatal wards, while KPC-KP predominated in ICUs, with KPC-KP showing higher antimicrobial resistance.

Objective:The performance of GeneXpert MTB/RIF (Xpert), DNA isothermal amplification fluorescence assay (DNA isothermal amplification) and RNA simultaneous amplification and testing (SAT-TB) were evaluated by detecting Mycobacterium tuberculosis in the pus samples of superficial tuberculous lymphadenitis and skeletal tuberculosis, two common extrapulmonary tuberculosis. Methods:Cross-sectional study. A total of 242 patients with suspected superficial tuberculous lymphadenitis and skeletal tuberculosis admitted to Shaanxi Provincial Tuberculosis Prevention and Control Hospital were collected from January 2022 to December 2023 and pus samples were taken from the lesions for examination. Among them, 210 patients were laboratory confirmed or clinically diagnosed with tuberculosis, 108 patients with superficial tuberculous lymphadenitis and 102 patients with skeletal tuberculosis. 32 patients without tuberculosis. Mycobacterium Tuberculosis culture, Xpert, DNA isothermal amplification, and SAT-TB detection were performed on the pus samples of all patients. The detection results were statistically analyzed using SPSS26.0 and MedCalc software. The detection effectiveness of several different detection methods for Mycobacterium tuberculosis in pus samples was compared, and P<0.05 was considered statistically significan. Results:The sensitivity of Mycobacterium Tuberculosis culture, Xpert, DNA isothermal amplification and SAT-TB in TB patients were 25.2% (53/210), 88.6% (186/210), 81.9% (172/210) and 61.0% (128/210), respectively. The area under curve (AUC) values of the four detection methods in the diagnosis of tuberculosis were 0.626, 0.943, 0.910 and 0.805, respectively. The AUC value of tuberculosis culture in the diagnosis of tuberculosis was significantly lower than that of the three molecular diagnostic techniques ( P<0.05). The AUC value of DNA isothermal amplification was significantly higher than that of SAT-TB detection method ( P<0.05). The positive tuberculosis culture was used as a reference standard where the sensitivity of Xpert, DNA isothermal amplification and SAT-TB reached 96.2%(51/53), 90.6%(48/53) and 86.8%(46/53), respectively. There was no significant difference in sensitivity among the three molecular diagnostic techniques ( P>0.05). In Xpert MTB positive patients, the positive rate of DNA isothermal amplification was 90.3% (168/186), and there was good consistency between DNA isothermal amplification and Xpert MTB detection results ( Kappa=0.765, P<0.001). Xpert detected 27 cases of rifampicin resistance, the resistance rate was 12.9% (27/210). Conclusion:GeneXpert and DNA isothermal amplification have high sensitivity and specificity in the samples of extrapulmonary tuberculosis pus, and the results of DNA isothermal amplification and GeneXpert MTB/RIF detection are highly consistent. In tuberculosis screening, DNA isothermal amplification fluorescence detection can be used for preliminary screening to reduce the detection cost.

Objective To investigate the changing antimicrobial resistance profiles of Streptococcus pneumoniae isolates from children in Chongqing area from 2011 to 2022,and to provide evidence for rational use of antibiotics and prevention of nosocomial infections.Methods The clinical data of S.pneumoniae strains isolated from pediatric patients during 2011-2022 were retrospectively analyzed.Antimicrobial susceptibility testing was performed with commercial automated systems and E-test.The results were interpreted according to the breakpoints in CLSI document(2022 edition).Results A total of 26 668 strains of S.pneumoniae were isolated during the 12-year period.The proportion of S.pneumoniae was 16.0％in the total pathogenic bacterial isolates and 46.4％in all the gram-positive bacterial pathogens.S.pneumoniae strains were mainly isolated from respiratory specimens(97.1％),followed by blood samples(1.5％).The highest proportion of S.pneumoniae isolates was in infants(38.2％),followed by toddlers(32.4％),preschool age(22.9％),school age(5.6％),adolescents(0.6％)and neonates(0.4％).All of the 38 strains of nonmeningitis SS.pneumoniae(0.1％)isolated from cerebrospinal fluid were resistant to penicillin.Overall,35.7％and 32.4％of these strains were resistant to cefotaxime and meropenem,respectively.The majority of S.pneumoniae(99.9％,26 630/26 668)were nonmeningitis isolates.The prevalence of penicillin-susceptible(PSSP),-intermediate(PISP),and-resistant(PRSP)strains was 71.9％(16 083),25.1％(5 610),and 3.0％(674),respectively.The prevalence of PRSP in infants and preschool children was higher than that in other age groups.The nonmeningitis S.pneumoniae isolates showed higher than 95％resistance rate to erythromycin,clindamycin and tetracycline,but 0.2％,0.2％and 0.1％resistance rate to levofloxacin,moxifloxacin and rifampicin,respectively.No S.pneumoniae strains were found resistant to vancomycin or linezolid.Conclusions The proportion and antimicrobial resistance profiles of S.pneumoniae strains isolated from pediatric patients varied with age group and specimen type.The decreasing prevalence of PRSP may inform empirical treatment of S.pneumoniae infections in children in Chongqing area.

Objective:To discuss the regulatory role of complement alternative pathway in mouse colorectal cancer(CRC)liver metastasis model based on bioinformatics methods,and to clarify its mechanism through experimental verification.Methods:Using"CRC liver metastasis"as the keyword,the GSE81558 dataset was retrieved from Gene Expression Omnibus(GEO)database,including normal colon tissue samples,CRC tissue samples and CRC liver metastasis tissue samples.Bioinformatics methods were used to analyze and screen differentially expressed genes(DEGs).Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed using R and Cytoscape software,and the results were visualized.Search Tool for the Retrieval of Interacting Genes/Proteins(STRING)database was used to evaluate protein-protein interactions(PPIs)of DEGs and construct PPI network.Twelve C57BL/6 mice were injected with SL4 tumor cells into spleen,and the liver tissues were collected at 0,7 and 14 d.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of complement pathway-related genes in liver metastatic foci.The CRC liver metastasis mouse model was used to verify the complement signaling pathway.The mice were divided into control group,factor B knockout group(FB-/-)and C4 factor knockout group(C4-/-),and there were 6 mice in each group.The liver weights of the mice were measured;HE staining was used to detect the percentage of metastatic area in liver tissue in control group and FB-/-group;immunohistochemistry was used to detect macrophage infiltration in liver tissue in control group and FB-/-group,and the percentage of macrophage infiltration was calculated.Results:The distances between normal colon tissue samples and CRC tissue samples,as well as between CRC tissue samples and CRC liver metastasis tissue samples were far,indicating significant differences between samples,allowing subsequent analysis of DEGs.A total of 1 908 DEGs were screened in the dataset comparing normal colon tissue samples and CRC tissue samples,including 771 up-regulated DEGs and 1 137 down-regulated DEGs.Twenty-three up-regulated DEGs and 100 down-regulated DEGs were identified in the dataset comparing CRC and CRC liver metastasis.The GO functional enrichment analysis results showed that compared with normal colon tissue samples,DEGs in CRC samples were mainly enriched in biological processes(BP)related to cell cycle and mitosis,including mitotic cell cycle process,cell division,response to hormone,mitotic nuclear division and response to lipid.Compared with CRC samples,the DEGs in CRC liver metastasis samples were mainly enriched in coagulation-related BP,including platelet degranulation,blood coagulation regulation,acute-phase response,hemostasis regulation and coagulation regulation.The KEGG pathway enrichment analysis results showed that compared with normal colon tissue samples,the DEGs in CRC tissue samples were mainly enriched in cell cycle and p53 signaling pathways.Compared with CRC tissue samples,the DEGs in CRC liver metastasis tissue samples were mainly enriched in complement,coagulation cascade and metabolism-related signaling pathways.The Hub genes identified in PPI network were related to blood proteins.The RT-qPCR results showed that compared with 0 d group,the mRNA expression level of complement related genes complement 1q(C1q)in liver metastatic foci tissue sampres in 7 d group was significantly decreased(P<0.05),the mRNA expression levels of complement 3(C3),complement 5(C5),FB,and factor D(FD)were significantly increased(P<0.05 or P<0.01),the mRNA expression levels of complement pathway-related genes C1q,complement 2(C2),C3,complement fragment 3a receptor(C3aR),C5,complement fragment 5a receptor(C5aR),decay-accelerating factor(DAF),FB and FD in liver metastatic foci tissue sampres in 14 d group were significantly increased(P<0.05 or P<0.01).Compared with control group,the liver weight of the mice in FB-/-group was significantly decreased(P<0.01),while there was no significant difference was observed in C4-/-group(P>0.05).The HE staining results showed that compared with control group,the liver metastatic foci in FB-/-mice were significantly decreased,and the percentage of metastatic area was decreased(P<0.01).The immunohistochemistry results showed that compared with control group,the macrophage infiltration in liver metastatic foci of the mice in FB-/-group was reduced,and the percentage of macrophage infiltration was decreased(P<0.01).Conclusion:Complement cascade is associated with CRC liver metastasis,and the alternative complement pathway regulates CRC liver metastasis,suggesting this pathway may serve as a potential therapeutic target for CRC liver metastasis.

Objective To investigate the changing antimicrobial resistance profiles of Streptococcus pneumoniae isolates from children in Chongqing area from 2011 to 2022,and to provide evidence for rational use of antibiotics and prevention of nosocomial infections.Methods The clinical data of S.pneumoniae strains isolated from pediatric patients during 2011-2022 were retrospectively analyzed.Antimicrobial susceptibility testing was performed with commercial automated systems and E-test.The results were interpreted according to the breakpoints in CLSI document(2022 edition).Results A total of 26 668 strains of S.pneumoniae were isolated during the 12-year period.The proportion of S.pneumoniae was 16.0％in the total pathogenic bacterial isolates and 46.4％in all the gram-positive bacterial pathogens.S.pneumoniae strains were mainly isolated from respiratory specimens(97.1％),followed by blood samples(1.5％).The highest proportion of S.pneumoniae isolates was in infants(38.2％),followed by toddlers(32.4％),preschool age(22.9％),school age(5.6％),adolescents(0.6％)and neonates(0.4％).All of the 38 strains of nonmeningitis SS.pneumoniae(0.1％)isolated from cerebrospinal fluid were resistant to penicillin.Overall,35.7％and 32.4％of these strains were resistant to cefotaxime and meropenem,respectively.The majority of S.pneumoniae(99.9％,26 630/26 668)were nonmeningitis isolates.The prevalence of penicillin-susceptible(PSSP),-intermediate(PISP),and-resistant(PRSP)strains was 71.9％(16 083),25.1％(5 610),and 3.0％(674),respectively.The prevalence of PRSP in infants and preschool children was higher than that in other age groups.The nonmeningitis S.pneumoniae isolates showed higher than 95％resistance rate to erythromycin,clindamycin and tetracycline,but 0.2％,0.2％and 0.1％resistance rate to levofloxacin,moxifloxacin and rifampicin,respectively.No S.pneumoniae strains were found resistant to vancomycin or linezolid.Conclusions The proportion and antimicrobial resistance profiles of S.pneumoniae strains isolated from pediatric patients varied with age group and specimen type.The decreasing prevalence of PRSP may inform empirical treatment of S.pneumoniae infections in children in Chongqing area.

Objective:To investigate the clinical distribution characteristics changes in antimicrobial resistance, and carbapenemase resistance genes of Klebsiella pneumoniae isolated from children in Chongqing region during the period of January 2019 to December 2024, providing a basis for the rational use of antibiotics and the prevention and control of nosocomial infections.Methods:An observational study was conducted to retrospectively analyze 5 020 Klebsiella pneumoniae (KP) isolates detected in four hospitals of the Southwest Pediatric Laboratory Specialty Alliance. Antimicrobial susceptibility testing was performed by the minimum inhibitory concentration method combined with the disk diffusion method. Results were interpreted according to the 2024 Clinical and Laboratory Standards Institute (CLSI) standards. Carbapenemase resistance genes were detected by polymerase chain reaction (PCR) combined with Sanger sequencing. WHONET 5.6 was used for resistance analysis and SPSS 19.0 for statistical analysis. The chi-square test was used to assess trends in resistance rates, ESBL detection rates, and resistance rates of different CRKP carbapenemase genotypes from 2019 to 2024. Statistical significance was confirmed if the two-tailed P-value was <0.05. Results:A total of 5 020 strains were isolated, with a detection rate of 5.1% (5 020/99 063). The majority were from sputum (59.2%, 2 970/5 020), followed by pus (17.1%, 857), urine (9.7%, 487), venous blood (6.5%, 326), secretions (2.6%, 130), and other specimens (5.0%, 250).The lowest resistance rate was to amikacin (3.8%), followed by levofloxacin (10.9%), imipenem (19.1%), and meropenem (19.9%). Resistance rates to cefoperazone/sulbactam ( χ2=9.982 0, P=0.001 6), piperacillin/tazobactam ( χ2=10.110 0, P=0.001 5), ceftazidime ( χ2=3.849 0, P=0.049 8), cefotaxime ( χ2=7.605 0, P=0.005 8), cefepime ( χ2=13.510 0, P=0.000 2), aztreonam ( χ2=6.457 0, P=0.011 1), imipenem ( χ2=4.672 0, P=0.030 7), and levofloxacin ( χ2=7.555 0, P=0.006 0) showed an annual increasing trend. The main carbapenemase genes were blaNDM-5 (42.2%, 127/301), blaNDM-1 (33.9%, 102/301), and blaKPC-2 (17.3%, 52/301). Patients with KPC-2-producing strains (median age, 240 days) were older than those with NDM-1/NDM-5-producing strains (median age, 40 days) ( χ2=22.620 0, P<0.000 1). In neonatal wards, the detection rate of NDM-KP was higher than that of KPC-KP (64.6%, 148/229 vs. 26.9%, 14/52, χ2=24.680 0, P<0.000 1), whereas in ICUs, it was lower (6.1%, 14/229 vs. 26.9%, 14/52, χ2=20.450 0, P<0.000 1). Conclusion:In Chongqing region, the isolation rate of K. pneumoniae from sputum was the highest with most cases from neonatal wards. Resistance to carbapenems showed an upward trend. BlaNDM-5 was the predominant genotype in pediatric CRKP. Patients with KPC-KP were older than those with NDM-KP. NDM-KP predominated in neonatal wards, while KPC-KP predominated in ICUs, with KPC-KP showing higher antimicrobial resistance.