Objective To explore the role of miR-429 in synovial mesenchymal stem cell-derived exosomes(SMSC-Exos)in repairing cartilage damage in temporomandibular joint osteoarthritis(TMJ OA)by extracting SMSC-Exos from human synovial tissue and screening differentially expressed microRNA(miRNA)through transcriptome sequencing.Methods Human synovial tissues were obtained from 6 patients who underwent surgery at the First Medical Center of the Chinese PLA General Hospital from June to December 2023,including 3 patients with osteoarthritis(OA group)and 3 control patients(control group),all of whom were male.SMSC-Exos were extracted from the synovial tissues for miRNA sequencing and differential expression analysis.Further,Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed on the target genes of differentially expressed miRNA to identify key functional miRNA and construct miRNA-target gene regulatory networks and protein-protein interaction(PPI)networks of target genes.An in vitro model of rabbit condylar cartilage cell inflammatory microenvironment induced by interleukin-1β(IL-1β)was established,with the control group cultured in DMEM/F12 basic medium and the inflammation-induced group cultured in DMEM/F12 basic medium containing 10 ng/ml IL-1β.RT-qPCR was used to detect the effects of overexpressed target miRNA on the mRNA expression levels of cartilage phenotype factors such as type Ⅱ collagen α1 chain(Col2a1),aggrecan(Acan),as well as inflammatory factors including a disintegrin and metalloproteinase with thrombospondin motifs 5(Adamts5)and cyclooxygenase-2(Cox-2).Results(1)SMSC-Exos were successfully isolated,cultured,and identified.(2)miRNA sequencing of SMSC-Exos from OA and control groups revealed 16 differentially expressed miRNAs(|log2FC|>2,P<0.05).Compared with control group,7 miRNAs were up-regulated and 9 were down-regulated in OA group.GO and KEGG analysis indicated that the target genes of miR-429 were mainly involved in development process,anatomical structure development,system development,cell development and differentiation,and were enriched in inflammation-related pathways such as mitogen-activated protein kinase(MAPK)and phosphatidylinositol 3-kinase-protein kinase B(PI3K-Akt).(3)Functional validation of miR-429 in the rabbit condylar cartilage cell inflammatory model showed that overexpression of miR-429 increased the mRNA expression levels of Col2a1 and Acan(P<0.05)and decreased the mRNA expression levels of Adamts5 and Cox-2(P<0.05)in the inflammation-induced group.Conclusions miRNA sequencing of SMSC-Exos isolated and identified from human synovial tissues reveals a specific miRNA expression profile in OA patients,with miR-429 significantly down-regulated.Functional validation demonstrates that overexpression of miR-429 has reparative and anti-inflammatory effects on condylar cartilage cells in an inflammatory microenvironment.

Objective To understand the distribution characteristics of fungi on object surface in hospital environ-ment,and provide reference for the scientific and precise formulation of environment control strategies based on fun-gal in clinic.Methods From December 7 to 23,2023,a total of 60 environmental specimens of 19 categories in 6 departments of a large tertiary first-class hospital were collected and divided into water-related environmental speci-men group,complete facade environmental specimen group,and sanitary ware environmental specimen group.18S rRNA sequencing was performed on specimens with fungi detected.Results Fungal detection rate of environmental specimens was 20.00％(12/60).Sink in the department of endocrinology had the highest fungal colony count(15 CFU/cm2),followed by the air outlet of air disinfection device in the department of thoracic surgery and the in-ternal part of a faucet in the department of endocrinology(both 10 CFU/cm2).The water-related environmental specimen group detected most diverse fungal genera(14 species),with high relative abundances of Aspergillus(100％),Meyerozyma(99.06％),Ophiocordyceps genus(95.63％),and Kodamaea(87.86％).The air outlet of air disinfection device was detected with a high abundance of Chaetomium(44.08％)and Corollospora(39.71％).There was no statistically significant difference in the α-diversity(Shannon and Simpson indices,P values of 0.661 and 0.568,respectively)and β-diversity(P=0.712)among the three environmental specimens.Conclusion Under the routine implementation of basic environmental cleaning and disinfection in medical institutions,fungi are in a low prevalence in the environment.However,moist surfaces and air disinfection device are prone to fungal colonization,and it is necessary to strengthen daily monitoring and take corresponding intervention measures to reduce the risk of infection.

Objective:To investigate the relationship between screen time and content, and the mental health status of adolescents. The findings will inform the formulation of targeted intervention policies to enhance adolescent mental health.Methods:Between September and November 2023, 5 197 students from 64 junior high, senior high, and vocational schools across 13 districts in Wuhan were recruited, using the stratified whole-cluster random sampling to investigate their screen behavior and mental health status. Mental health status was measured using the Mental Health Inventory for Chinese Middle School Students (MMHI-60). A generalized additive model was used to explore the nonlinear association between screen time and mental health status. Additionally, a mixed-effects model was utilized to explore the dose-response associations between average daily total screen time, screen time for different content types, and adolescents' mental health status and the impact of the proportion of different screen contents on mental health outcomes.Results:The age of the participants was (14.40±1.48) years, with 56.07% being boys. The MMHI-60 score averaged 1.73±0.70. The M( Q1,Q3) for daily total screen time was 50.00 (0.00,128.57) minutes. The M( Q1,Q3) for screen time dedicated to gaming, studying, socializing, and watching videos were 0.00 (0.00, 20.00), 8.57 (1.64, 44.50), 4.28 (0.00, 30.00), and 0.00 (0.00, 25.71) minutes, respectively. A non-linear association was observed between average daily screen time and adolescent mental health problem score, 0-1 hour of daily screen time was beneficial for adolescent mental, compared to no screen time. However, screen time exceeding 1 hour was detrimental, with the negative impact increasing alongside screen time duration. When total daily screen time was held constant, the proportion of time spent on gaming ( β=0.14, 95% CI: 0.05-0.23, P=0.003) and video ( β=0.21, 95% CI: 0.09-0.28, P<0.001) was positively correlated with mental health problems, whereas the proportion of time spent on studying was negatively correlated with mental health problems ( β=-0.17, 95% CI: -0.24 - -0.11, P<0.001). Conclusions:Moderate screen time is advantageous for adolescent mental health. However, it is crucial to minimize the proportion of screen time dedicated to video and gaming activities to mitigate potential adverse effects.

Objective: To investigate the nutritional status and influencing factors among Tibetan and Mongolian children and adolescents aged 7-18 years in high-altitude regions, so as to provide evidence for early prevention and control of malnutrition in this population. Methods: From May to June 2022, a cluster sampling method was employed to recruit 1 019 Tibetan and Mongolian children and adolescents aged 7-18 years from two primary and secondary schools in Golmud City. Physical examinations, dietary frequency questionnaires, and physical activity assessments were conducted. Nutritional status was classified as obesity, combined overweight/obesity, underweight, or central obesity according to national standards including Screening for Overweight and Obesity among School-age Children and Adolescents, Screening Standard for Malnutrition of School-age Children and Adolescents, Blue Book on Obesity Prevention and Control in China. Chi-square tests, t-test and Logistic regression analyses were performed to identify factors associated with different nutritional statuses. Results: The detection rates of obesity, combined overweight/obesity, underweight, and central obesity were 8.0%, 18.1%, 5.2%, and 19.7%, respectively. The height of children and adolescents across all age groups was generally lower than the national standard values. Tibetan participants exhibited significantly lower height-for-age Z-scores (HAZ)(9-10, 13-17 years, Z =2.01, 2.78, 4.16, 3.38, 4.12, 3.63, 3.00) and BMI-for-age Z-scores (BAZ) compared to Mongolian participants ( Z =-2.95, -2.47, -2.31, -2.89, -2.14, -2.17)( P < 0.05 ). Multivariate Logistic regression revealed that Mongolian children and adolescents had higher risks of obesity ( OR =2.20) and combined overweight/obesity ( OR = 2.18 ) ( P <0.05). Additionally, insufficient moderate-to-vigorous physical activity (MVPA) was associated with an increased risk of central obesity ( OR =1.48, P <0.05), compared with children and adolescents who meet the standard of MVPA. Conclusions The rates of overweight and obesity among Tibetan and Mongolian children and adolescents in Golmud City are higher, influenced by multiple factors. Nutrition interventions and physical activity strategies tailored to ethnic characteristics should be implemented, with emphasis on promoting MVPA to improve nutritional outcomes in this population.

Objective:To investigate the relationship between screen time and content, and the mental health status of adolescents. The findings will inform the formulation of targeted intervention policies to enhance adolescent mental health.Methods:Between September and November 2023, 5 197 students from 64 junior high, senior high, and vocational schools across 13 districts in Wuhan were recruited, using the stratified whole-cluster random sampling to investigate their screen behavior and mental health status. Mental health status was measured using the Mental Health Inventory for Chinese Middle School Students (MMHI-60). A generalized additive model was used to explore the nonlinear association between screen time and mental health status. Additionally, a mixed-effects model was utilized to explore the dose-response associations between average daily total screen time, screen time for different content types, and adolescents' mental health status and the impact of the proportion of different screen contents on mental health outcomes.Results:The age of the participants was (14.40±1.48) years, with 56.07% being boys. The MMHI-60 score averaged 1.73±0.70. The M( Q1,Q3) for daily total screen time was 50.00 (0.00,128.57) minutes. The M( Q1,Q3) for screen time dedicated to gaming, studying, socializing, and watching videos were 0.00 (0.00, 20.00), 8.57 (1.64, 44.50), 4.28 (0.00, 30.00), and 0.00 (0.00, 25.71) minutes, respectively. A non-linear association was observed between average daily screen time and adolescent mental health problem score, 0-1 hour of daily screen time was beneficial for adolescent mental, compared to no screen time. However, screen time exceeding 1 hour was detrimental, with the negative impact increasing alongside screen time duration. When total daily screen time was held constant, the proportion of time spent on gaming ( β=0.14, 95% CI: 0.05-0.23, P=0.003) and video ( β=0.21, 95% CI: 0.09-0.28, P<0.001) was positively correlated with mental health problems, whereas the proportion of time spent on studying was negatively correlated with mental health problems ( β=-0.17, 95% CI: -0.24 - -0.11, P<0.001). Conclusions:Moderate screen time is advantageous for adolescent mental health. However, it is crucial to minimize the proportion of screen time dedicated to video and gaming activities to mitigate potential adverse effects.

Objective To understand the distribution characteristics of fungi on object surface in hospital environ-ment,and provide reference for the scientific and precise formulation of environment control strategies based on fun-gal in clinic.Methods From December 7 to 23,2023,a total of 60 environmental specimens of 19 categories in 6 departments of a large tertiary first-class hospital were collected and divided into water-related environmental speci-men group,complete facade environmental specimen group,and sanitary ware environmental specimen group.18S rRNA sequencing was performed on specimens with fungi detected.Results Fungal detection rate of environmental specimens was 20.00％(12/60).Sink in the department of endocrinology had the highest fungal colony count(15 CFU/cm2),followed by the air outlet of air disinfection device in the department of thoracic surgery and the in-ternal part of a faucet in the department of endocrinology(both 10 CFU/cm2).The water-related environmental specimen group detected most diverse fungal genera(14 species),with high relative abundances of Aspergillus(100％),Meyerozyma(99.06％),Ophiocordyceps genus(95.63％),and Kodamaea(87.86％).The air outlet of air disinfection device was detected with a high abundance of Chaetomium(44.08％)and Corollospora(39.71％).There was no statistically significant difference in the α-diversity(Shannon and Simpson indices,P values of 0.661 and 0.568,respectively)and β-diversity(P=0.712)among the three environmental specimens.Conclusion Under the routine implementation of basic environmental cleaning and disinfection in medical institutions,fungi are in a low prevalence in the environment.However,moist surfaces and air disinfection device are prone to fungal colonization,and it is necessary to strengthen daily monitoring and take corresponding intervention measures to reduce the risk of infection.

Objective To explore the influence of extracellular matrix protein ABI-interactor 3-binding protein(ABI3BP)on vesicular stomatitis virus(VSV)genome replication and innate immune signaling pathway. Methods The small interfering RNA(siRNA)was transfected to knock down ABI3BP gene in human skin fibroblast BJ-5ta cells.VSV-green fluorescent protein(VSV-GFP)-infected cell model was established.The morphological changes and F-actin stress fiber formation were detected on ABI3BP knockdown cells by phalloidin immunofluorescence staining.The mRNA level of virus replication was detected by RT-qPCR in BJ-5ta cells after VSV-GFP infection;western blotting was performed to detect the changes in interferon regulatory factor 3(IRF3)andTANK-binding kinase 1(TBK1)phosphorylation levels. Results The VSV-GFP-infected BJ-5ta cell model was successfully established.Efficient knockdown of ABI3BP in BJ-5ta cells was achieved.Phalloidin immunofluorescence staining revealed structural rearrangement of intracellular F-actin after ABI3BP gene knockdown.Compared with the control group,the gene copy number of VSV-GFP in ABI3BP knockdown cells increased by 2.2-3.5 times(P＜0.01)and 2.2-4.0 times(P＜0.01)respectively when infected with VSV of multiplicity of infection 0.1 and 1.The expression of viral protein significantly increased in ABI3BP knockdown cells after virus infection.The activation of type-Ⅰ interferon pathway,as determined by phosphorylated IRF3 and phosphorylated TBK1,was significantly decreased in ABI3BP knockdown cells after VSV-GFP infection. Conclusions Extracellular matrix protein ABI3BP plays an important role in maintaining the formation and rearrangement of actin structure.ABI3BP gene deletion promotes RNA virus replication,and ABI3BP is an important molecule that maintains the integrity of type Ⅰ interferon pathway.

Objective To identify related factors for hypoglycemic episodes in patients with type 2 diabetes mellitus(T2DM)through continuous glucose monitoring(CGM). Methods The included 147 patients with T2DM were those who had undergone CGM for 5 days in our ward of Department of Endocrinology and Metabolism,Huashan Hospital from Dec 2018 to Oct 2019. The general information, laboratory parameters and CGM parameters of the patients were collected. According to whether there wasan episode of hypoglycemia during the monitoring period,the patients were divided into non-hypoglycemia group and hypoglycemic group. A single hypoglycemia episode was defined as a sensor monitoring blood glucose of less than 3.9 mmol/L and lasting for more than 15 minutes.CGM parameters included the mean blood glucose(MBG),standard deviation(SD),coefficient of variation(CV),the differences between maximum and minimum blood glucose (BG) levels (ΔBG),mean amplitude of glycemic excursions (MAGE)and the percentage of time in range(％TIR)of BG at <3.9 mmol/L,3.9-7.8 mmol/L,>7.8 mmol/L,3.9-10.0 mmol/L,and >10.0 mmol/L. Results Logistic regression analysis showed that lower estimated glomerular filtration rate(eGFR)levels,increased use of insulin and its analogs and lower MBG levels were associated with hypoglycemic episodes. Spearman correlation analysis showed that the MBG level and the ％TIR of BG>7.8 mmol/L and BG>10.0 mmol/L were negatively associated while glycemic variability(GV)levels(SD,CV,ΔBG,MAGE)and ％ TIR of BG at 3.9-7.8 mmol/L were positively associated with hypoglycemic episodes. Pearson correlation analysis showed that the duration of hypoglycemic episodes was positively correlated with the use of sulfonylureas and CV levels. Conclusion Lower eGFR levels,increased treatment with insulin and its analogs and lower MBG levels were related factors for hypoglycemic episodes in patients with T2DM.

The capacity for thermal tolerance is critical for industrial enzyme. In the past decade, great efforts have been made to endow wild-type enzymes with higher catalytic activity or thermostability using gene engineering and protein engineering strategies. In this study, a recently developed SpyTag/SpyCatcher system, mediated by isopeptide bond-ligation, was used to modify a rumen microbiota-derived xylanase XYN11-6 as cyclized and stable enzyme C-XYN11-6. After incubation at 60, 70 or 80 ℃ for 10 min, the residual activities of C-XYN11-6 were 81.53%, 73.98% or 64.41%, which were 1.48, 2.92 or 3.98-fold of linear enzyme L-XYN11-6, respectively. After exposure to 60-90°C for 10 min, the C-XYN11-6 remained as soluble in suspension, while L-XYN11-6 showed severely aggregation. Intrinsic and 8-anilino-1-naphthalenesulfonic acid (ANS)-binding fluorescence analysis revealed that C-XYN11-6 was more capable of maintaining its conformation during heat challenge, compared with L-XYN11-6. Interestingly, molecular cyclization also conferred C-XYN11-6 with improved resilience to 0.1-50 mmol/L Ca²⁺ or 0.1 mmol/L Cu²⁺ treatment. In summary, we generated a thermal- and ion-stable cyclized enzyme using SpyTag/SpyCatcher system, which will be of particular interest in engineering of enzymes for industrial application.
Animals ; Cyclization ; Endo-1,4-beta Xylanases ; chemistry ; metabolism ; Enzyme Stability ; Industrial Microbiology ; methods ; Microbiota ; Protein Engineering ; Rumen ; enzymology ; microbiology ; Temperature

Objective: To explore the lipid-lowering effect and potential mechanism of the couplet medicines of Flastem Milkvetch Seed and Tribulus terrestris on hyperlipidemia rats. Methods: Forty-eight Sprague-Dawley rats were randomly divided into 6 groups, normal control group, high-fat model group, simvastatin group, Tribulus terrestris group, Flastem Milkvetch Seed group, and couplet medicines of Flastem Milkvetch Seed and Tribulus terrestris group, with 8 rats in each group. The normal control group was fed with basal diet, and the other groups were fed with high-fat diet to establish a hyperlipidemia rat model. At the same time, different group rats were treated with different drug respectively. 6 weeks later, the body weight and visceral index of the rats were measured. Levels of serum cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), and low density lipoprotein cholesterol (LDL-C) were detected by automatic biochemical analyzer. The mRNA and protein expression of HMG coenzyme A reductase (HMGCR), cholesterol 7-alpha hydroxy-lase (CYP7A1) and low density lipoprotein receptor (LDL-R) were detected by RT-PCR and western blot. Results: Compared with model group, the serum levels of TC, TG, and LDL-C were significantly decreased in all Chinese medicine groups, and the serum level of HDL-C was significantly increased in Flastem Milkvetch Seed group and couplet medicines of Flastem Milkvetch Seed and Tribulus terrestris group. Moreover, the mRNA and protein expression of HMGCR, CYP7A1, and LDL-R in rat liver also significantly increased in all Chinese medicine treatment groups. And couplet medicines of Flastem Milkvetch Seed and Tribulus terrestris group shows more significantly effect. Conclusion: Couplet medicines of Flastem Milkvetch Seed and Tribulus terrestris significantly regulate the serum lipid levels on hyperlipidemia rat model, and its mechanism may be related to the regulating expressions of HMGCR, CYP7A1, LDL-R gene.