Objective:To analyze the efficacy and safety of Minocycline for treating macrolide-unresponsive Mycoplasma pneumoniae pneumonia (MUMPP) in children aged 6-14 years, and to investigate the influencing factors of severe Mycoplasma pneumoniae pneumonia (SMPP). Methods:Retrospective cohort study.The clinical data of 247 children with MUMPP hospitalized in the Department of Pediatrics, the First Affiliated Hospital of Anhui Medical University from August to December 2023 were analyzed.According to the medication 3 to 7 days after conventional macrolide treatment, the children were divided into an Azithromycin group (107 cases) and a Minocycline group (140 cases). Clinical indicators such as the length of hospital stay and chest radiograph absorption time were compared between the two groups.Multivariate Logistic regression analysis was performed on children with SMPP to explore the correlation of the incidence of severe cases with the initiation time of Minocycline, serum vitamin D, lactate dehydrogenase (LDH), C-reactive protein (CRP) levels.Results:In the Minocycline group, the median hospital stay was shortened by 1.0 d (4.0 d vs.5.0 d, P=0.008), the median time for cough improvement after hospitalization was reduced by 3.0 d (2.0 d vs.5.0 d, P<0.001), and the duration of coughing, time to normal body temperature, and the 1-2-week improvement rate of chest imaging [91.4%(128/140) vs.49.5%(53/107), P<0.001] were all significantly better, compared with those in the Azithromycin group.In addition, the Glucocorticoid use rate [35.7%(50/140) vs.49.5%(53/107), P=0.029] in the Minocycline group was significantly lower than that in the Azithromycin group.The incidence of drug-related adverse reactions was only 0.71%(1/140) in the Minocycline group.A longer duration of macrolide treatment before Minocycline administration [odds ratio ( OR)=1.386, 95% confidence interval ( CI): 1.038-1.850, P=0.027], lower serum vitamin D levels ( OR=0.841, 95% CI: 0.748-0.945, P=0.004), elevated LDH levels ( OR=1.023, 95% CI: 1.009-1.037, P=0.001), and elevated CRP levels ( OR=1.074, 95% CI: 1.035-1.114, P<0.001) were associated with SMPP.The area under the receiver operating characteristic curve of the combined model for predicting SMPP was 0.912 7. Conclusions:Minocycline shows better efficacy than Azithromycin in treating MUMPP children aged 6-14 years, with comparable safety.LDH and CRP levels are independent risk factors for SMPP, while the serum vitamin D level is a protective factor.Early initiation of Minocycline treatment can optimize the clinical outcome.

OBJECTIVE To explore the effect of fluconazole on proteomics of Trichosporon asahii so as to reveal the responding process of T.asahii to fluconazole stress and the resistance mechanisms to azoles on the protein level.METHODS T.asahii AS 2.2174 was chosen as the research subject,the minimum inhibitory concentration(MIC)of fluconazole was determined by broth microdilution assay.The protein abundance of T.asahii was detec-ted by means of tandem mass tag(TMT)technique combined with liquid chromatography-tandem mass spectrom-etry(LC-MS/MS)before and after the treatment with fluconazole(1× MIC).The differentially expressed pro-teins(DEPs)were identified based on the screening standards of fold change ≥1.20 or ≤0.83 and P＜0.05.Gene ontlogy(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis were performed for the DEPs so as to understand the biological property of the DEPs and the major biological pathways that the DEPs in-volved in.Finally,the targeted validation was carried out for the targeted differentially expressed proteins by using multiple reaction monitoring(MRM).RESULTS The MIC of fluconazole to T.asahii AS 2.2174 was 8 μg/ml.Totally 196 DEPs were identified,including 93 upregulated DEPs and 103 downregulated DEPs.The function en-richment analysis showed that the DEPs mainly participated in synthesis and metabolism of sterols,drug metabo-lism,stress response,energy metabolism and intertranslation.The targeted DEPs showed the consistent expres-sion trends in MRM target validation and TMT-LC-MS/MS.CONCLUSIONS The protein abundance of T.asahii has remarkable change under the fluconazole stress.The bioinformatics analysis reveals the complicated molecular mechanisms of T.asahii in response to the fluconazole stress,which may offer valuable ideas for understanding the drug resistance to azoles and developing new drug targets.

Objective To isolate pathogenic bacteria from the skin of a nude mouse exhibiting squamous skin scurfs, and perform bacterial identification, traceability analysis, and pathogenicity studies to provide a new approach for the diagnosis of pathogens in nude mice with squamous skin scurfs. MethodsSkin swab samples were collected from a nude mouse exhibiting squamous skin scurfs for nucleic acid testing, bacterial isolation and culture, biochemical identification, 16S rDNA gene amplification and sequencing, and whole genome sequencing to construct a phylogenetic tree. Fifteen BALB/c nude mice were randomized into a saline-treated control group, a high-concentration group treated with 1.8×10⁸ CFU/mL of the isolated bacterial suspension, and a low-concentration group treated with 1.8×10⁷ CFU/mL of the isolated bacterial suspension. Pathogenicity was assessed by animal infection experiments and observation of histopathological changes in skin tissue using HE staining. Results The nucleic acid test for Corynebacterium bovis was negative, excluding infection by this organism. The pathogen isolated on mannitol salt agar and blood agar, combined with Gram staining, suggested a Gram-positive Staphylococcus species. The isolated strain was identified by 16S rDNA sequencing and a fully automated microbial identification system as Staphylococcus xylosus. Phylogenetic tree analysis based on whole genome sequencing showed that the strain was most closely related to an isolate from leafy vegetables in South Korea (GenBank GCA_00207825.1). In the high-concentration group, squamous skin scurfs appeared on the head, neck, and back of nude mice on the 17th day post-infection, while in the low concentration group, similar symptoms appeared on the 20th day post-infection and gradually spread to other areas. The scaling symptoms were transient, lasting for 7 days in the high-concentration group and 3 days in the low-concentration group, after which the skin returned to normal. The infection rate was 33.33% in both the high- and low-concentration groups. No significant pathological changes were observed in the skin tissues of infected mice compared to the control group, indicating marked individual differences in the pathogenicity of the strain in nude mice. Conclusion A strain of Staphylococcus xylosus was isolated from the skin of a nude mouse exhibiting squamous skin scurfs. The strain is an opportunistic pathogen that causes transient squamous skin scurfs without significant histopathological changes, and there are individual differences in the sensitivity of nude mice to this strain. These findings can provide valuable data for pathogen identification in immunodeficient or gene knockout mice.

Objective To isolate pathogenic bacteria from the skin of a nude mouse exhibiting squamous skin scurfs, and perform bacterial identification, traceability analysis, and pathogenicity studies to provide a new approach for the diagnosis of pathogens in nude mice with squamous skin scurfs. MethodsSkin swab samples were collected from a nude mouse exhibiting squamous skin scurfs for nucleic acid testing, bacterial isolation and culture, biochemical identification, 16S rDNA gene amplification and sequencing, and whole genome sequencing to construct a phylogenetic tree. Fifteen BALB/c nude mice were randomized into a saline-treated control group, a high-concentration group treated with 1.8×10⁸ CFU/mL of the isolated bacterial suspension, and a low-concentration group treated with 1.8×10⁷ CFU/mL of the isolated bacterial suspension. Pathogenicity was assessed by animal infection experiments and observation of histopathological changes in skin tissue using HE staining. Results The nucleic acid test for Corynebacterium bovis was negative, excluding infection by this organism. The pathogen isolated on mannitol salt agar and blood agar, combined with Gram staining, suggested a Gram-positive Staphylococcus species. The isolated strain was identified by 16S rDNA sequencing and a fully automated microbial identification system as Staphylococcus xylosus. Phylogenetic tree analysis based on whole genome sequencing showed that the strain was most closely related to an isolate from leafy vegetables in South Korea (GenBank GCA_00207825.1). In the high-concentration group, squamous skin scurfs appeared on the head, neck, and back of nude mice on the 17th day post-infection, while in the low concentration group, similar symptoms appeared on the 20th day post-infection and gradually spread to other areas. The scaling symptoms were transient, lasting for 7 days in the high-concentration group and 3 days in the low-concentration group, after which the skin returned to normal. The infection rate was 33.33% in both the high- and low-concentration groups. No significant pathological changes were observed in the skin tissues of infected mice compared to the control group, indicating marked individual differences in the pathogenicity of the strain in nude mice. Conclusion A strain of Staphylococcus xylosus was isolated from the skin of a nude mouse exhibiting squamous skin scurfs. The strain is an opportunistic pathogen that causes transient squamous skin scurfs without significant histopathological changes, and there are individual differences in the sensitivity of nude mice to this strain. These findings can provide valuable data for pathogen identification in immunodeficient or gene knockout mice.

OBJECTIVES: This study aimed to use machine learning algorithms to build a prediction model of the first permanent molar caries of 9-year-old children in Suzhou and screen out risk factors. METHODS: Random stratified whole group sampling was applied to randomly select 9-year-old students from 38 primary schools in 14 townships and streets in Wuzhong District for oral examination and questionnaire survey. Multifactor Logistics regression was used to analyze the risk factors of tooth decay. The data set was randomly divided into training sets and verification sets according to 8∶2, and R 4.3.1 was used to build five machine learning algorithms: random forest, decision tree, extreme gradient boosting (XGBoost), Logistics regression, and lightweight gradient enhancement (LightGBM). The predictive effect of these five models was evaluated using the area under the characteristic curve (AUC). The marginal contribution of quantitative characteristics to the caries prediction model was determined through Shapley additive explanations (SHAP). RESULTS: This study included 7 225 samples that met the standard. The caries rate of the first permanent molar was 54.96%. Multifactor Logistic regression analysis showed that sweet drinks, dessert and candy, snack frequency, and snacks before going to bed after brushing teeth were correlated with the occurrence of first permanent molar caries (P<0.05). The AUC values of decision tree, Logistic regression, LightGBM, random forest, and XGBoost were 75.5%, 83.9%, 88.6%, 88.9%, and 90.1%, respectively. Compared with the variables after single heat coding, the SHAP value of high-frequency sweets (such as dessert candy ≥2 times a day, mother's sugary diet ≥2 times a day) and bad oral hygiene habits (such as frequent snacks before going to bed after brushing teeth and irregular brushing teeth) exhibited the highest positive. CONCLUSIONS XGBoost algorithm has a good prediction effect for first permanent molar caries in 9-year-old children. High-frequency sweet factors and bad oral hygiene habits have a strong positive impact on the risk of first permanent molar caries and are key drivers that can be used in the formulation of targeted interventions.
Humans ; Dental Caries/epidemiology* ; Child ; Machine Learning ; China/epidemiology* ; Molar ; Risk Factors ; Female ; Logistic Models ; Male ; Decision Trees ; Algorithms

Objective To study the action of Sijunzi decoction preventing disuse atrophy and its mechanism on the pathway of muscle stem cell(MuSC)/myonuclear domain(MND)based on the theory of sedentary behavior damaging muscle.Methods 40 male rats were randomly divided into the normal group,tail hanging group(model group),tail hanging+exercise group(exercise group),tail hanging+distilled water group(DW group),tail hanging+sijunzi decoction(Chinese medicine(CM)group);grip strength of limbs was measured by Grip Strength Meter for rats and mice;the wet weight of quadriceps femoris(QF)was measured by electronic scales;concentrations of actin and myosin were detected by ELISA;the cross section area(CSA)and myonuclear numbers were detected with Image Pro 6.0,and then MNDs were calculated;the expressions of pair box gene 7(Pax7)and myogenic differentiation antigen(MyoD)were detected by immunofluorescence.Results Compared with those in the normal group,grip strength,QF wet weight,the myosin concentration,the CSA and MND in the model and DW groups were all decreased,the numbers of Pax7 were increased significantly;compared with those in the model and distilled water groups,grip strength,QF wet weight,the myosin level,CSA and myonuclear numbers in the exercise group and CM groups were increased,and the numbers of Pax7 and MyoD positive cells were increased,markedly,however the number of Pax7 positive cells in the exercise group was more than that in the CM group significantly.Conclusion Sijunzi decoction can prevent disuse atrophy and its possible mechanism is related to the activation the MuSC/MND pathway.

Objective:To analyze the efficacy and safety of Minocycline for treating macrolide-unresponsive Mycoplasma pneumoniae pneumonia (MUMPP) in children aged 6-14 years, and to investigate the influencing factors of severe Mycoplasma pneumoniae pneumonia (SMPP). Methods:Retrospective cohort study.The clinical data of 247 children with MUMPP hospitalized in the Department of Pediatrics, the First Affiliated Hospital of Anhui Medical University from August to December 2023 were analyzed.According to the medication 3 to 7 days after conventional macrolide treatment, the children were divided into an Azithromycin group (107 cases) and a Minocycline group (140 cases). Clinical indicators such as the length of hospital stay and chest radiograph absorption time were compared between the two groups.Multivariate Logistic regression analysis was performed on children with SMPP to explore the correlation of the incidence of severe cases with the initiation time of Minocycline, serum vitamin D, lactate dehydrogenase (LDH), C-reactive protein (CRP) levels.Results:In the Minocycline group, the median hospital stay was shortened by 1.0 d (4.0 d vs.5.0 d, P=0.008), the median time for cough improvement after hospitalization was reduced by 3.0 d (2.0 d vs.5.0 d, P<0.001), and the duration of coughing, time to normal body temperature, and the 1-2-week improvement rate of chest imaging [91.4%(128/140) vs.49.5%(53/107), P<0.001] were all significantly better, compared with those in the Azithromycin group.In addition, the Glucocorticoid use rate [35.7%(50/140) vs.49.5%(53/107), P=0.029] in the Minocycline group was significantly lower than that in the Azithromycin group.The incidence of drug-related adverse reactions was only 0.71%(1/140) in the Minocycline group.A longer duration of macrolide treatment before Minocycline administration [odds ratio ( OR)=1.386, 95% confidence interval ( CI): 1.038-1.850, P=0.027], lower serum vitamin D levels ( OR=0.841, 95% CI: 0.748-0.945, P=0.004), elevated LDH levels ( OR=1.023, 95% CI: 1.009-1.037, P=0.001), and elevated CRP levels ( OR=1.074, 95% CI: 1.035-1.114, P<0.001) were associated with SMPP.The area under the receiver operating characteristic curve of the combined model for predicting SMPP was 0.912 7. Conclusions:Minocycline shows better efficacy than Azithromycin in treating MUMPP children aged 6-14 years, with comparable safety.LDH and CRP levels are independent risk factors for SMPP, while the serum vitamin D level is a protective factor.Early initiation of Minocycline treatment can optimize the clinical outcome.

Objective To study the action of Sijunzi decoction preventing disuse atrophy and its mechanism on the pathway of muscle stem cell(MuSC)/myonuclear domain(MND)based on the theory of sedentary behavior damaging muscle.Methods 40 male rats were randomly divided into the normal group,tail hanging group(model group),tail hanging+exercise group(exercise group),tail hanging+distilled water group(DW group),tail hanging+sijunzi decoction(Chinese medicine(CM)group);grip strength of limbs was measured by Grip Strength Meter for rats and mice;the wet weight of quadriceps femoris(QF)was measured by electronic scales;concentrations of actin and myosin were detected by ELISA;the cross section area(CSA)and myonuclear numbers were detected with Image Pro 6.0,and then MNDs were calculated;the expressions of pair box gene 7(Pax7)and myogenic differentiation antigen(MyoD)were detected by immunofluorescence.Results Compared with those in the normal group,grip strength,QF wet weight,the myosin concentration,the CSA and MND in the model and DW groups were all decreased,the numbers of Pax7 were increased significantly;compared with those in the model and distilled water groups,grip strength,QF wet weight,the myosin level,CSA and myonuclear numbers in the exercise group and CM groups were increased,and the numbers of Pax7 and MyoD positive cells were increased,markedly,however the number of Pax7 positive cells in the exercise group was more than that in the CM group significantly.Conclusion Sijunzi decoction can prevent disuse atrophy and its possible mechanism is related to the activation the MuSC/MND pathway.

OBJECTIVE To explore the effect of fluconazole on proteomics of Trichosporon asahii so as to reveal the responding process of T.asahii to fluconazole stress and the resistance mechanisms to azoles on the protein level.METHODS T.asahii AS 2.2174 was chosen as the research subject,the minimum inhibitory concentration(MIC)of fluconazole was determined by broth microdilution assay.The protein abundance of T.asahii was detec-ted by means of tandem mass tag(TMT)technique combined with liquid chromatography-tandem mass spectrom-etry(LC-MS/MS)before and after the treatment with fluconazole(1× MIC).The differentially expressed pro-teins(DEPs)were identified based on the screening standards of fold change ≥1.20 or ≤0.83 and P＜0.05.Gene ontlogy(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis were performed for the DEPs so as to understand the biological property of the DEPs and the major biological pathways that the DEPs in-volved in.Finally,the targeted validation was carried out for the targeted differentially expressed proteins by using multiple reaction monitoring(MRM).RESULTS The MIC of fluconazole to T.asahii AS 2.2174 was 8 μg/ml.Totally 196 DEPs were identified,including 93 upregulated DEPs and 103 downregulated DEPs.The function en-richment analysis showed that the DEPs mainly participated in synthesis and metabolism of sterols,drug metabo-lism,stress response,energy metabolism and intertranslation.The targeted DEPs showed the consistent expres-sion trends in MRM target validation and TMT-LC-MS/MS.CONCLUSIONS The protein abundance of T.asahii has remarkable change under the fluconazole stress.The bioinformatics analysis reveals the complicated molecular mechanisms of T.asahii in response to the fluconazole stress,which may offer valuable ideas for understanding the drug resistance to azoles and developing new drug targets.

Objective:To evaluate the epidemiological characteristics and explore the associated factors of breakthrough cases (BC) from Public Health Emergency Events (PHEEs) of varicella in Zhejiang Province from 2019 to 2022.Methods:Data on cases were obtained from the China Information System for Disease Control and Prevention and the PHEEs Reporting Information Database of Varicella in Zhejiang Province. History records were matched through the Zhejiang Provincial Immunization Information System. Descriptive analysis and multiple logistic regression model with a bidirectional stepwise selection method were performed to explore associated factors for BC during 2019-2022.Results:A total of 144 276 varicella cases were reported from 2019 to 2022, with the annual reported incidence of 47.35-82.80 cases per 100 000 population. Among these cases, 109 172 were non-breakthrough cases (NBC, accounting for 75.67%), 34 517 were BC (23.92%), and the rest 587 cases had unclear vaccination history on varicella (0.41%). A total of 214 PHEEs of varicella were reported, of which 99.07% occurred in school settings. The proportion of PHEEs that occurred in high school increased significantly as time went on ( χ2trend=5.742, P=0.017). Multiple logistic regression model which focused on "BC vs. NBC (as the reference)" indicated that the year of onset ( OR=1.585, 95% CI:1.343-1.878), the month of onset (taking January as the reference, OR=2.311-15.652), city (taking Hangzhou as the reference, Jiaxing OR=2.370, Jinhua OR=2.197, Lishui OR=0.134), age ( OR=0.887, 95% CI: 0.826-0.944), PHEEs setting (taking "primary school and below" as the reference, "high school and above" OR=0.516, 95% CI: 0.305-0.897), and the number of rashes ( OR=0.569, 95% CI: 0.458-0.703) were associated factors. Multiple logistic regression model which focused on "two-dose BC vs. one-dose BC (as the reference)" showed that the age of initial vaccination ( OR=0.045, 95% CI: 0.014-0.107), the time interval from onset to the last dose ( OR=0.037, 95% CI: 0.011-0.087) and the age of onset ( OR=20.724, 95% CI: 8.383-72.485) were associated factors. Conclusion:During 2019-2022, the reported high-risk group of varicella in Zhejiang Province has shifted to adolescents and young adults. Although vaccination could not completely prevent the onset of VZV, it could relieve clinical symptoms and delay the age of onset.