Objective To investigate the inhibitory effect of methylprednisolone (MP) on the development of coronary arteritis in a murine model of Kawasaki disease (KD) induced with a candida albicaus watersoluble fraction (CAWS).Methods Forty-five C57BIL/6mice were evenly divided into three groups (the control group,the CAWS group and the MP group).Mice in the CAWS group were intraperitoneally injected phosphate buffer saline (PBS) for 5 days.MP or PBS was administered to the different group.The animals were sacrificed at day 3,day 10 and day 28,and the status of vasculitis in the coronary arteries and the aortic root was investigated histologically.One-way analysis of variance (ANOVA) was used to compare the differences among three groups,and t-test for two independent groups.Results The mice in CAWS group and MP group,which induced by CAWS,showed that the body weight and heart weight decreased significantly,and the spleen weight was increased at day 10 and day 28 (P＜0.05).Vasculitis was induced in the mice of those two groups,and the severity score was the highest at day 10 (12.7±0.5).In addition,the severity of the inflammation of the aortic root and the coronary arteries were reduced in MP group (t=6.35,5.55,2.8,P＜0.05).Elastic fiber staining showed that the layers of vascular walls were in disorder and elastic fibers were broken in the CAWS group.However,there was no disruption or breakage in the MP group.Conclusion MP can suppress the progression of coronary arteritis in this CAWS-induced murine vasculitis model,which indicates the efficacy of MP in KD patients with coronary artery lesions.

Objective To study the distribution and drug resistance of pathogens in neonatal septicemia in order to provide clinical guidance for antibiotic usage.Methods This retrospective study analyzed blood culture and clinical data from 83 confirmed neonatal septicemia patients and the blood collection cultures were analyzed.Results A total of 84 strains were isolated from 83 cases of blood specimens,Gram positive bacteria,Gram negative bacteria and fungi were 38(45.2%),41(48.8%),5(6.0%),respectively.Gram positive bacteria was mainly coagulase negative staphylococcus and staphylococcus aureus,which were 13(15.5%) and 8(9.5%) respectively.Gram negative bacteria was mainly Escherichia coli and Klebsiella pneumonia,which were 25(29.8%) and 9(10.7%) respectively.Gram positive bacteria were found high resistance to penicillin G,amoxicillin clavulanate potassium,oxacillin and clindamycin,from 34.2% to 73.7%,but they were sensitive to vancomycin,teicoplanin and linezolid.Gram negative bacteria were found high resistance to ampicillin(82.9%),the constituent ratio of the extended spectrum βlactamases(ESBLs) was 34.1%,carbapenem resistant strains was not found.All fungi were sensitive to azoles.Conclusion Gram negative bacteria are the major pathogens in neonatal septicemia,with high infection rate of Escherichia coli and high constituent ratio of the ESBLs,and antimicrobial agents should be chosen according to blood culture and antimicrobial susceptibility results.

Objective To investigate the expression of interleukin-35 (IL-35) in serum of mice with pulmonary interstitial fibrosis.Methods Thirty-six C57BL/6 mice were divided into three groups (twelve mice in each group):control group,model group of mice with bleomycin-induced pulmonary fibrosis,glucocorticoid treatment group of mice with bleomycin-induced pulmonary fibrosis.The mice were sacrificed at day 7,day 14 and day 28 respectively,and the serum concentration of IL-35 was assayed.Statistical product and service solutions (SPSS) 17.0 statistical software was used for single factor analysis of variance and LSD-t comparison and Pearson correlation analysis was used for the comparison between each two groups.Results The serum IL-35 concentrations between groups and within groups at the time of day 7,day 14 and day 28 were compared respectively.The serum IL-35 concentration of the model group was significantly lower than the control group and the glucocorticoid treatment group at the time of day 7 (F=24.56,P＜0.05).The serum IL-35 concentration of glucocorticoid treatment group was significantly lower than the control group at the time of day 14 (F=8.85,P＜0.05),and which of glucocorticoid treatment group was significantly lower than the control group and the model group at the time of day 28 (F=36.64,P＜0.05).There was no significant difference between days 28 and day 7 within control group (t=1.03,P＞0.05).The serum IL-35 concentration of the model group at the time of day 28 was significantly higher than those of day 7 [(9.36±0.95) ng/ml vs (6.51±0.58) ng/ml,t=5.14,P＜0.05],and which of glucocorticoid treatment group was significantly lower than those of day 7 [(5.27±1.01) ng/ml vs (9.42±0.84) ng/ml,t=6.32,P＜0.05].From day 7 to day 28 the serum IL-35 concentration change of the model group and glucocorticoid treatment group showed significantly negative correlation (r =-0.743,P＜0.05).Conclusion Serum IL-35 concentration has shown an trend of increase during bleomycin-induced pulmonary fibrosis with mice model,and early glucocorticoid treatment can decrease the serum IL-35 in the model mice.

Objective To explore the effect of different concentrations of iguratimod (IGU) on mouse model of bleomycin-induced pulmonary fibrosis.Methods A total of 108 female C57BL/6 mice were randomly divided into the control group,the model group,the low dose IGU group,the moderate dose IGU group,high dose with group and the methylprednisolone (MP) group (n=18 in each group).The mice in the control group were injected with 0.2 ml normal saline endotracheally,and others were injected with 0.2 ml bleomycin (5 mg/kg) from endotracheally to induce pulmonary fibrosis model.The next day,the mice in both control group and the model group were fed with 0.2 ml normal saline every day;The mice in the IGU groups and methylprednisolone group were fed with 0.2 ml iguratimod liquid the IGU (10,30,90 mg/kg) and 0.2 ml methylprednisolone (10 mg/kg) every dayrespectively.Finally the mice were sacrificed at day 7,day 14,day 28 respectively,and the lung tissue was examined by HE staining and Masson staining to evaluate the degree of alveolitis and fibrosis.Repeated measurement of variance analysis was used to compare the differences for time and group,and multi-factor analysis of variance LSD test was used for the comparison between groups.Results ① The body mass of mice in bleomycin-induced groups were decreased compared to the control group.② The pathological alveolitis scores in the high dose IGU group and methylprednisolone group were significantly decreased compared to those of the model IGU group at day 7 and day 14 (P＜0.05),and the pathological fibrosis scores were decreased dramatically compared with the model group at day 14 and day 28 (P＜0.05).Conclusion High concentration of IGU and methylprednisolone can reduce and inhibit lung inflammation and fibrosis of bleomycin-induced pulmonary fibrosis in mice.

Objective:To establish a method for the determination of aflatoxin( AF) B1,B2,G1and G2by HPLC in Xiao'er Fupi granule,and help manufacturers select safe raw materials for production through the analysis of test results of 31 batches of samples and the safety investigation of Xiao'er Fupi granule in terms of aflatoxin pollution. Methods: A post-column photochemical derivation HPLC method was used to detect the content of aflatoxin in Xiao'er Fupi granule. An Ecosil C18column(250 mm×4.6 mm,5 μm) was adopted with the mobile phase of acetonitrile-methanol-water (30: 10: 60)at the flow rate of 0.8 ml·min-1. The column tem-perature was maintained at 30 ℃. The sample size was 20 μl,The excitation wavelength was 360 nm and the emission wavelength was 450 nm in the fluorescence detection. Results:The linear range of aflatoxin B1,B2, G1and G2was 9.65-48.25 pg (r=0.999 1), 2.45-12.25 pg(r=0.999 8), 10.5-52.5 pg(r =0.995 6) and 2.55-12.75 pg (r =0.996 6), respectively. The recovery was 83.3%-95.6%. Totally 14 batches of samples contained aflatoxin,and the total content was 0.21 × 10 -3-0.54 ×10 -3μg·g-1. Conclusion:The method is convenient and accurate,which can be used for the quality control of Xiao'er Fupi granule.

Objectives: To explore the differences in signal pathway and gene expression related to the pathogenesis of congenital microtia by the in-depth analysis of DNA methylation profiling of auricular chondrocytes from congenital microtia patients. Methods: Genome wide methylation profile of congenital microtia was obtained by MeDIP chip technology, and analyzed by Gene ontology (GO) and Pathway analysis. The gene expression levels of Wnt1 and Wnt11 were evaluated by Real-time PCR in the auricular cartilage from the healthy side and affected side of the congenital microtia patients , and healthy controls. Results: The GO and Pathway assay showed that Wnt signal pathway was enriched in differential methylated levels. The Wnt1 and Wnt11 genes were with higher methylation in the promoter region and CpG islands in healthy control group than that in microtia group, in addition the methylation level in the affected side auricular cartilage was lower than that in the healthy side. There was no difference in Wnt1 and Wnt11 gene expression in microtia patients and healthy controls. The higher Wnt11 gene expression was detected in the affected side residual cartilage tissues than in the healthy side cartilage tissues of the same congenital microtia patient. Conclusions The over expression of Wnt11 during embryonic development might be associated with the pathogenesis of congenital microtia. The mechanism of the difference in methylation levles of Wnt11 affecting pathogenesis of congenital microtia needs further research.

Objective:To explore the characteristics of plasma microRNA (miRNA) profiles and bioinformatics in patients with osteoarthritis(OA) in order to search for diseases related biomarkers.Methods:Blood samples from 20 cases of OA patients and 15 cases of normal control (NC) were collected to extracted total RNA in plasma. The plasma miRNA expression profile was tested by using Agilent Human miRNA array. Target gene analysis and clustering analysis were performed on differentially expressed microRNAs. Three differentially expressed miRNAs (miR-134-5p, miR-320c and miR-940) were detected by real-time quantitative polymerase chain reaction (RT-qPCR) for further confirmation of microarray data. The differences were tested using t test analysis. Results:① MiRNA microarray showed that compared with NC, there were 74 differential expression genes in plasma of patients in the OA group (FC≥2, P≤0.01), among which 45 were up-regulated and 29 were down-regulated. ② A total of 2 731 potential target genes were predicted in three database, and involved in 462 Kyoto Encyclopedia of Genes and Genomes (KEEG) pathways. Target gene ontology (GO) functional clustering found that the main functions of miRNAs were intercellular adhesion, collagen synthesis, intracellular signal transduction, etc. The main KEGG pathways of miRNAs include mitogen-activated protein kinase (MAPK) signaling pathway, cyclic adenosine monophosphate (cAMP) signaling pathway, osteoclast differentiation signaling pathway, etc. ③ The expression level of miR-20a-5p and miR-320c in OA group were significantly higher than that in controls ( t=-6.142, P<0.05; t=-3.854, P<0.05), while miR-940 was significantly lower than that of controls ( t=2.767, P<0.05) . The trend was consistent with the microarray data. The receiver operating characteristic curve (ROC) curve analyses showed that they were useful biomarkers for differentiating patients with OA from controls. Conclusion:The study shows that plasma in OA patients has a specific miRNAs expression, and miRNAs play an important role in the pathogenesis of OA.

Objective:To investigate the expression level of plasma miR-320c in patients with osteoarthritis(OA), and explore the clinical significance and the role in pathogenesis of OA.Methods:The clinical data and peripheral blood of 30 patients with OA, 30 patients with connective tissue diseases and 30 healthy control individuals were collected.The levels of plasma miR-320c were detected byfluorescentquantitative reverse transcription PCR(qRT-PCR). Correlation analysis was used to explore the correlation of plasma miR-320c level with knee X-ray data and VAS pain score in OA patients.Finally the miR-320c mimic, the miR-320c inhibitor, and the control material were transfected to the chondrocyte HC-a.The proliferative capacity of HC-a chondrocytes was examined at different time points as determined by the CCK-8 assay.Results:The expression level of plasma miR-320c was significantly higher in OA group(3.26±0.55)than that in the connective tissue diseases group(1.62±0.50)and in healthy control group(1.21±0.66)( F=107.66, P<0.001). Plasma miR-320c expression was positively correlated with radiographic grade( r=0.830, P<0.001), and had no correlation with VAS pain score in OA group( P>0.05). Through repeated measurement variance analysis, the time effect, the group effect and the interaction effect between group and time showed statistically significant differences in chondrocyte proliferation between NC mimic group and miR-320c mimic group( Ftime=5256.767, Fgroup=1947.436, Ftime×group=114.314, all P<0.001). The level of proliferation was significantly reduced.Apoptosis rate of chondrocytes was significantly increased in the group transfected with miR-320c( t=7.85, P<0.01). Conclusions:The expression level of plasma miR-320c is significantly higher in osteoarthritis patients and associated with knee radiographic severity grade.Furthermore, over-expression of miR-320c could suppress the proliferation of chondrocytes.Plasma miR-320c might be potential bio-marker for osteoarthritis knee severity assessment, and involves in regulating chondrocyte growth in the pathogenesis of osteoarthritis.

Objective:To investigate the epidemiology and clinical characteristics of adenovirus (ADV)-caused acute respiratory tract infection among hospitalized children in Kunming, China.Methods:Clinical and laboratory data were collected from 467 children with adenovirus infection who were hospitalized from January 1, 2019 to December 31, 2019 in 6 grade A class Ⅲ hospitals in Kunming area. The basic characteristics, epidemiology, mixed infection and adenovirus genotypes of the patients were retrospectively analyzed. The patients diagnosed with adenovirus pneumonia (AP) were divided into two groups, severe AP (SAP) group and general AP(GAP) group according to the severity of illness. Mann-Whitney U test or χ 2 test was used for comparison between groups, while multivariate regression was applied to analyze the risk factors of SAP. Results:Among 15 635 hospitalized children with respiratory tract infection, 467 cases were adenovirus positive, with a detection rate of 2.99%. Of the 467 patients with adenovirus infection, 284 were male and 183 female, the age was 2.4 (1.1,3.9) years, including 44 cases (9.4%) < 0.5 years, 59 cases (12.6%) of 0.5 to<1.0 years, 176 cases (37.7%) of 1.0 to <3.0 years, 150 cases (32.1%) of 3.0 to <7.0 years, and 38 cases (8.1%) of 7.0 to 14.0 years. Adenovirus infection was common in autumn and winter, and the high incidence months were October to December, which accounted for 51.6% (241/467) of the whole year cases. Co-infection was detected in 226 cases (48.4%) out of 467 patients, in which one pathogen co-infection was the most frequent form (172 cases, 76.1%). Of the 262 pathogen detected 108 (41.2%) were Mycoplasma pneumoniae. In 144 of ADV-positve cases (30.8%) were taken geno-typing was done by PCR amplification, the results showed that 74 cases (51.4%) were ADV 3, 7 subtypes and 65 cases (45.1%) of ADV 1, 2,6 subtypes. Of the 467 cases of ADV infection, 320 (68.5%) were diagnosed with pneumonia, 82 (17.6%) with upper respiratory tract infection and pharyngeal tonsillitis, and 65 (13.9%) with bronchitis, laryngeal bronchitis, and asthmatic bronchitis. Among the 320 patients with AP, 56 cases were severe and 264 cases were general. Two cases (3.6%) in severe group died. Compared with the GAP group, the age was young [17 (11,42) months vs. 24 (14,44) months, Z=2.222, P=0.026], the fever duration was long [8 (5,14) days vs. 6 (3,9) days, Z=3.380, P<0.01], and the proportions of preterm birth and having underlying diseases were high [respectively 19.6% (11/56) vs. 6.1% (16/264), 26.8% (15/56) vs. 10.2% (27/264), χ 2=8.965,11.109, P<0.05] in SAP group. Referring to laboratory markers, white blood cell count, C-reactive protein, creatine kinase-MB and lactate dehydrogenase were significantly increased in SAP group as compared to GAP group(all P<0.05). Multivariate Logistic regression analysis showed that preterm birth ( OR=3.284, 95% CI 1.079-9.993, P=0.036), underlying disease ( OR=3.284, 95% CI 1.079-9.993, P=0.036), fever duration ≥10 d ( OR=2.523,95% CI 1.195-5.328, P=0.015) and C-reactive protein ≥50 mg/L ( OR=3.156, 95% CI 1.324-7.524, P=0.010) were positively correlated with the risk of SAP. Conclusions:The incidence of adenovirus infection among hospitalized children in Kunming was lower than the national level, and no outbreak occurred in 2019. Subtype 3 and 7 of ADV are the predominant strains for infection, which usually occurs in autumn and winter and mainly causes pneumonia. Premature birth, underlining diseases, long fever duration and markedly increased C-reactive protein are the risk factors for developing into severe pneumonia. This paper presents the prevalence and clinical characteristics of adenovirus infection in children at high altitude area.

Objective:To compare the epidemiological and clinical characteristics of hospitalized children with respiratory syncytial virus (RSV) infection in Kunming among the pre-and post-COVID-19 era, and to establish a prediction model for severe RSV infection in children during the post-COVID-19 period.Methods:This was a retrospective study. Clinical and laboratory data were collected from 959 children hospitalized with RSV infection in the Department of Pulmonary and Critical Care Medicine at Kunming Children′s Hospital during January to December 2019 and January to December 2023. Patients admitted in 2019 were defined as the pre-COVID-19 group, while those admitted in 2023 were classified as the post-COVID-19 group. Epidemiological and clinical characteristics were compared between the two groups. Subsequently, comparison of the clinical severity among the two groups was performed based on propensity score matching (PSM). Furthermore, the subjects in the post-COVID-19 group were divided into severe and non-severe groups based on clinical severity. Chi-square test and Mann-Whitney U test were used for pairwise comparison between groups, and multivariate Logistic regression was applied for the identification of independent risk factors and construction of the prediction model. The receiver operating characteristic (ROC) curve and calibration curve were employed to evaluate the predictive performance of this model. Results:Among the 959 children hospitalized with RSV infection, there were 555 males and 404 females, with an onset age of 15.4 (7.3, 28.5) months. Of which, there were 331 cases in the pre-COVID-19 group and 628 cases in the post-COVID-19 group. The peak period of RSV hospitalization in the post-COVID-19 group were from May to October 2023, and the monthly number of inpatients for each of these months were as follows: 72 cases (11.5%), 98 cases (15.6%), 128 cases (20.4%), 101 cases (16.1%), 65 cases (10.4%), and 61 cases (9.7%), respectively. After PSM for general data, 267 cases were matched in each group. The proportion of wheezing in the post-COVID-19 group was lower than that in the pre-COVID-19 group (109 cases (40.8%) vs. 161 cases (60.3%), χ2=20.26, P<0.001), while the incidences of fever, tachypnea, seizures, severe case, neutrophil-to-lymphocyte ratio (NLR), C-reactive protein and interleukin-6 levels were all higher than those in the pre-COVID-19 group (146 cases (54.7%) vs. 119 cases (44.6%), 117 cases (43.8%) vs. 89 cases (33.3%), 37 cases (13.9%) vs. 14 cases (5.2%), 69 cases (25.8%) vs. 45 cases (16.9%), 3.6 (1.9, 6.4) vs. 2.3 (1.8, 4.6), 9.9 (7.1, 15.2) vs. 7.8 (4.5, 13.9) mg/L, 20.5 (15.7, 30.4) vs. 17.2 (11.0, 26.9) ng/L, χ2=5.46, 6.36, 11.47, 6.42, Z=4.13, 3.06, 2.96, all P<0.05). There were 252 cases and 107 cases with co-infection in the post-and pre-COVID-19 groups, respectively. The proportion of triple and quadruple infection in the post-COVID-19 group was higher than that in the pre-COVID-19 group (59 cases (23.4%) vs. 13 cases (12.1%), 30 cases (11.9%) vs. 5 cases (4.7%), χ2=5.94, 4.46, both P<0.05). Among the 252 cases with co-infection in post-COVID-19 group, the most prevalent pathogens involving in co-infections, in order, were Mycoplasma pneumoniae 56 cases (22.2%), Influenza A virus 53 cases (21.0%), Rhinovirus 48 cases (19.0%), Parainfluenza virus 35 cases (13.9%), and Adenovirus 28 cases (11.1%).The result of multivariate Logistic regression showed that age ( OR=0.70, 95% CI 0.62-0.78, P<0.001), underlying diseases ( OR=10.03, 95% CI 4.10-24.55, P<0.001), premature birth ( OR=6.78, 95% CI 3.53-13.04, P<0.001), NLR ( OR=1.85, 95% CI 1.09-3.15, P=0.023), and co-infection ( OR=1.28, 95% CI 1.18-1.38, P<0.001) were independently associated with the development of severe RSV infection in the post-COVID-19 group. The ROC curve of the prediction model integrating the above five factors indicated an area under the curve of 0.85 (95% CI 0.80-0.89, P<0.001), with an optimal cutoff of 0.21, a sensitivity of 0.83 and a specificity of 0.80. The calibration curve showed that the predicted probability in this model did not differ significantly from the actual probability ( P=0.319). Conclusions:In the post-COVID-19 era in Kunming, the peak in pediatric hospitalizations for RSV infection was from May to October, with declined incidence of wheezing and increased incidence of fever, tachypnea, seizures, severe cases, and rates of triple and quadruple co-infections. Age, underlying diseases, premature birth, NLR, and co-infection were identified as independent risk factors for severe RSV infection in the post-COVID-19 period. In this study, a risk prediction model for severe pediatric RSV infection was established, which had a good predictive performance.