Objective:To identify and validate secreted effector proteins of Chlamydia psittaci ( C. psittaci) through bioinformatic prediction and experimental verification, and to characterize their subcellular localization in host cells. Methods:Potential effector proteins were predicted using bioinformatics tools. Candidate effectors were fused to β-lactamase through the constructed expression vectors, and these vectors were transformed into C. psittaci. The secretion of these candidate effectors was evaluated by β-lactamase translocation assays. Eukaryotic expression vectors of confirmed effectors were transfected into host cells to determine their intracellular localization patterns. Results:Bioinformatic analysis identified 29 candidate effector proteins. Experimental validation confirmed the secretion of five effectors, with four exhibiting cytoplasmic localization and one displaying nuclear localization in host cells.Conclusion:This study characterizes five novel C. psittaci secreted effector proteins, providing critical insights for investigating the molecular pathogenesis of psittacosis.

Objective:To explore and evaluate the effectiveness of the new "D 3-teaching" mode in surgical training for ophthalmology standardized residency training. Methods:A total of 40 residents at the Eye Center of The Second Affiliated Hospital of Zhejiang University School of Medicine who participated in the standardized residency training program in Zhejiang Province from September 2020 to June 2024 were selected as the "D 3-teaching" group. This group received an innovative surgical training mode integrating digital resources, three-dimensional teaching, and dry lab simulation. Moreover, another 42 residents who had completed the standardized training program in the 2018 and 2019 were selected as the traditional teaching group. This group received traditional teaching methods involving theoretical lectures, text- and image-based teaching, and video demonstrations. The effectiveness of both teaching modes was assessed by comparing the pass rates of examinations, ICO-OSCAR skill mastery rates, learning periods, and satisfaction survey. Results:After training, there was no significant difference in the pass rates between the two groups. However, the skill mastery rate in the "D 3-teaching" group was significantly higher than that of the traditional teaching group (28/40, 70.00% vs. 15/42, 35.71%, χ2=9.66, P=0.002). Additionally, the learning period in the "D 3-teaching" group was significantly shorter than that of the traditional teaching group [(13.14±5.91) months vs. (21.33±7.73) months, t=3.88, P<0.001]. Furthermore, the scores for satisfaction with all teaching parameters in the "D 3-teaching" group were significantly higher than those in the traditional teaching group. Conclusions:The "D 3-teaching" mode is an innovative surgical teaching method in ophthalmology standardized residency training that significantly improves trainees' skill acquisition, enhances training outcomes, shortens the learning period, and has received high recognition from trainees. This mode is worthy of wider application and promotion.

Objective:To explore and evaluate the effectiveness of the new "D 3-teaching" mode in surgical training for ophthalmology standardized residency training. Methods:A total of 40 residents at the Eye Center of The Second Affiliated Hospital of Zhejiang University School of Medicine who participated in the standardized residency training program in Zhejiang Province from September 2020 to June 2024 were selected as the "D 3-teaching" group. This group received an innovative surgical training mode integrating digital resources, three-dimensional teaching, and dry lab simulation. Moreover, another 42 residents who had completed the standardized training program in the 2018 and 2019 were selected as the traditional teaching group. This group received traditional teaching methods involving theoretical lectures, text- and image-based teaching, and video demonstrations. The effectiveness of both teaching modes was assessed by comparing the pass rates of examinations, ICO-OSCAR skill mastery rates, learning periods, and satisfaction survey. Results:After training, there was no significant difference in the pass rates between the two groups. However, the skill mastery rate in the "D 3-teaching" group was significantly higher than that of the traditional teaching group (28/40, 70.00% vs. 15/42, 35.71%, χ2=9.66, P=0.002). Additionally, the learning period in the "D 3-teaching" group was significantly shorter than that of the traditional teaching group [(13.14±5.91) months vs. (21.33±7.73) months, t=3.88, P<0.001]. Furthermore, the scores for satisfaction with all teaching parameters in the "D 3-teaching" group were significantly higher than those in the traditional teaching group. Conclusions:The "D 3-teaching" mode is an innovative surgical teaching method in ophthalmology standardized residency training that significantly improves trainees' skill acquisition, enhances training outcomes, shortens the learning period, and has received high recognition from trainees. This mode is worthy of wider application and promotion.

Objective:To identify and validate secreted effector proteins of Chlamydia psittaci ( C. psittaci) through bioinformatic prediction and experimental verification, and to characterize their subcellular localization in host cells. Methods:Potential effector proteins were predicted using bioinformatics tools. Candidate effectors were fused to β-lactamase through the constructed expression vectors, and these vectors were transformed into C. psittaci. The secretion of these candidate effectors was evaluated by β-lactamase translocation assays. Eukaryotic expression vectors of confirmed effectors were transfected into host cells to determine their intracellular localization patterns. Results:Bioinformatic analysis identified 29 candidate effector proteins. Experimental validation confirmed the secretion of five effectors, with four exhibiting cytoplasmic localization and one displaying nuclear localization in host cells.Conclusion:This study characterizes five novel C. psittaci secreted effector proteins, providing critical insights for investigating the molecular pathogenesis of psittacosis.

Objective To isolate a Acinetobacter baumannii(Ab)phage from underground sewage,study its prop-erties,and to provide a theoretical basis for phage treatment of Ab infection.Methods Double-layer agar tech-nique was used to isolate phages by using Ab GY-6 as the host strain.Biological characterization and therapeutic effect of the phage was tested.Genetic information of the phage was analyzed.Results Ab phage Abgy202162 was isolated.Transmission electron microscopy(TEM)analysis showed that the morphology of Abgy202162 exhibited an icosahedral structure.Biological characteristic analysis showed that the optimal multiplicity of infection was 1,the latent period was 5 min,and the burst size was approximately 520 PFU per cell.In addition,Abgy202162 re-mained stable at different concentrations of chloroform,pH,and temperatures.Sodium dodecyl sulfate-polyacryl-amide gel electrophoresis(SDS-PAGE)analysis showed that it contained 10 proteins with molecular weights ran-ging from 15 to 100 ku.The double-stranded(ds)DNA genome of Abgy202162 consisted of 40 889 bp and its G+C content was 38.85％.It contained 47 open reading frames(ORFs),of which 26 had specific functions,but no virulence related genes or antibiotic resistance genes were found.Phylogenetic analysis showed that Abgy202162 was a new phage in the Autographiviridae family,Beijerinkvirinae subfamily,and Friunavirus genus.Abgy202162 showed the ability to prevent Ab infection in the Galleria mellonella in vivo model.Conclusion The phage Ab-gy202162 has strong environmental tolerance and high safety,indicating its potential as an antibiotic alternative used in the treatment of infections caused by Ab.

Objective To examine the mediating role of empathy and emotional competence in the association between family functioning and internalizing and externalizing problem behaviors among adolescents in China.Methods In this study,we used the data from the June-July 2022 survey of Chengdu Positive Child Development(CPCD)cohort.All respondents were 5th-9th graders from six primary or secondary schools in Chengdu.The Achenbach Child Behavior Checklist(CBCL),the Chinese Family Assessment Instrument(C-FAI),the empathy subscale of the Chinese version of the Interpersonal Reactivity Index(C-IRI),and the emotional competence(EC)subscale of the Chinese Positive Youth Development Scale(CPYD)were used to evaluate the respondents'internalizing and externalizing problem behaviors,family functioning,empathy,and emotional competence,respectively.The average score derived from the total score of a scale divided by the number of entries in each dimension was used as the final score of the scale.Independent samples t-tests or one-way analysis of variance(ANOVA)were performed to examine the differences in family functioning,empathy,emotional competence,and internalizing and externalizing problem behaviors across student groups with different demographic characteristics(sex,grade,and region).Pearson correlation analysis was conducted to examine the relationship between family functioning,empathy,emotional competence,and internalizing and externalizing problem behaviors.AMOS 24.0 was used to validate the hypothesized model and structural equation modeling was used to analyze the mediating effects of empathy and emotional competence between family functioning and internalizing and externalizing problem behaviors among adolescents.Results A total of 3026 eligible participants were included,including 1548(51.16%)male students and 1478(48.84%)female students.Among the respondents,798(26.37%)were 5th graders,738(24.39%)were 6th graders,567(18.74%)were 7th graders,614(20.29%)were 8th graders,and 309(10.21%)were 9th graders.In addition,2064(68.21%)of all respondents were from urban areas and 962(31.79%)were from rural areas.The results of the difference analysis showed that the differences in adolescents'internalizing and externalizing problem behaviors were statistically significant between students of different grades(P=0.004),and that the differences in family functioning and empathy scores were also statistically significant between students of different grades(all P<0.001),whereas the differences in adolescents'internalizing and externalizing problem behaviors were not statistically significant between sexes and regions(P=0.919,0.959).The results of correlation analysis showed that family functioning scores(the higher the score,the worse the family functioning)were significantly negatively correlated with empathy and emotional competence(r=-0.482,-0.432,P<0.01),and significantly positively correlated with internalizing and externalizing problem behaviors(r=0.220,P<0.01).Empathy was significantly positively correlated with emotional competence(r=0.402,P<0.01).Empathy and emotional competence were significantly negatively correlated with all the dimensions of internalizing and externalizing problem behaviors(r=-0.115,-0.305,P<0.01).Emotional competence partially mediated the relationship between family functioning and adolescents'internalizing and externalizing problem behaviors,with a mediation effect value being 0.042(95%[confidence interval]CI:0.031-0.057).Empathy and emotional competence had chain mediation effect between family functioning and adolescents'internalizing and externalizing problem behaviors,with the value of the mediation effect being 0.010(95%CI:0.007-0.014).Conclusion Family functioning influences adolescents'internalizing and externalizing problem behaviors in a direct way and through the chain-mediating roles of empathy and emotional competence.

Objective To investigate the effect of empathy on depressive symptoms in adolescents and to explore the potential mediating role of family functioning in the effect of empathy on depressive symptoms.Methods The 2022 cross-sectional data from the Chengdu Positive Child Development(CPCD)cohort were analyzed in the study.A survey was conducted in Chengdu in June 2022,involving 3 020 students in grades 5-8 from three randomly selected stratified schools.The Interpersonal Reactivity Index(IRI-C),the Chinese Family Assessment Instrument(C-FAI),and the Center for Epidemiologic Studies Depression Scale for Children(CES-DC)were used in the survey.Chi-square test or one-way analysis of variance was performed to examine the differences in various demographic characteristics(sex,grade,region,and total monthly household income)between groups of respondents,as well as the differences in family functioning,empathy,and depression.Pearson correlation coefficient was used to examine the correlation between family functioning,empathy,and depressive symptoms.Structural equation modeling and SPSS PROCESS component Model 4 were used to analyze whether family functioning played a mediating role in the effect of empathy on depressive symptoms in adolescents.Results The detection rate of depressive symptoms among survey respondents was 25.40%.The results of the difference analysis revealed significant differences in the detection rates of depressive symptoms among respondents of different grades,regions,and monthly household incomes(P<0.05).There was no significant difference in the detection rates of depressive symptoms between male and female students.There was a significant difference in the detection rate of depressive symptoms between respondents with different scores for family dysfunction and empathy ability(P<0.001).Correlation analysis results showed that empathy scores were negatively correlated with depression(r=-0.11,P<0.001),that family dysfunction was positively correlated with depression(r=0.29,P<0.001),and that empathy scores were negatively correlated with family functioning(r=-0.37,P<0.001).The mediating role of family dysfunction in the relationship between empathy and depressive symptoms was established,with the direct effect being 0.039(95% confidence interval[CI]:0.010-0.069,P<0.001)and the indirect effect value being-0.096(95% CI:-0.115--0.079,P<0.001).The direct effect value accounted for 28.89% of the total effect value,while the mediation effect value accounted for 71.11% of the total effect value.Conclusion The empathy ability of adolescents is correlated to depressive symptoms,and family functioning plays a mediating role between empathy and depressive symptoms in adolescents.It is suggested that adolescents'empathy ability and family functioning should be enhanced through multiple channels to reduce the occurrence of depressive symptoms.

OBJECTIVES: Hyperhomocysteinaemia (Hcy) is an independent risk factor for cardiovascular and cerebrovascular diseases. MicroRNA (miR)-18a-5p is closely related to cardiovascular diseases. This study aims to investigate the effects of miR-18a-5p on homocysteine (Hcy)-induced myocardial cells injury. METHODS: H9c2 cells were transfected with miR-18a-5p mimic/miR-18a-5p mimic negative control (NC) or combined with Hcy for intervention, and untreated cells were set as a control group. The transfection efficiency was verified by real-time RT-PCR, and cell counting kit-8 (CCK-8) assay was used to determine cell viability. Flow cytometry was used to detect apoptosis and reactive oxygen species (ROS) levels. Western blotting was performed to measure the protein levels of microtubule-associated protein 1 light chain 3 (LC3)-I, LC3-II, Beclin1, p62, Bax, Bcl-2, and Notch2. Dual luciferase reporter assay was used to detect the interaction of miR-18a-5p with Notch2. RESULTS: Compared with the control, treatment with Hcy or transfection with miR-18a-5p mimic alone, or combined treatment with Hcy and miR-18a-5p mimic/miR-18a-5p mimic NC significantly reduced the H9c2 cell viability, promoted apoptosis and ROS production, up-regulated the expressions of Bax and Beclin, down-regulated the expressions of Bcl-2, p62, and Notch2, and increased the ratio of LC3-II/LC3-I (all P<0.05). Compared with the combined intervention of miR-18a-5p mimic NC and Hcy group, the above indexes were more significantly changed in the combined intervention of miR-18a-5p mimic and Hcy group, and the difference between the 2 groups was statistically significant (all P<0.05). There is a targeted binding between Notch2 and miR-18a-5p. CONCLUSIONS MiR-18a-5p could induce autophagy and apoptosis via increasing ROS production in cardiomyocytes, and aggravate Hcy-induced myocardial injury. Notch2 is a target of miR-18a-5p.
Apoptosis/genetics* ; Autophagy/genetics* ; bcl-2-Associated X Protein ; MicroRNAs/metabolism* ; Proto-Oncogene Proteins c-bcl-2/genetics* ; Reactive Oxygen Species ; Rats ; Animals ; Myocytes, Cardiac/drug effects* ; Homocysteine/adverse effects* ; Hyperhomocysteinemia

Objective : To explore the role of Argonaute ( Ago) gene and RNA⁃Dependent RNA Polymerase (RDRP) gene of Aspergillus flavus in the growth and development about the RNAi mechanism . Methods : A. flavus Ago1 , Ago2 , RDRP1 , RDRP3 gene mutant strains were constructed by homologous recombination . The growth and development of the mutant strains were observed on potato dextrose agar(PDA) + uracil uridine (UU) medium inoculated with 3 μl 106 CFU/mL spores . 200 , 400 μg cell wall pressure agent conidored ( CR) , 0. 8 mol/L , 1 . 6 mol/L osmotic pressure agent NaCl , 2 mmol/L , 4 mmol/L oxidative pressure agent hydrogen peroxide (H2 O2 ) and 0. 01% , 0. 02% genomic damage agent methyl mesylate (MMS) were added to the Yeast extract Glucose Minimum (YGM) + UU medium to analyze the stress response of the mutant strains . Results : A. flavus mutant strains about ΔAgo1 , ΔAgo2 , ΔRDRP1 , ΔRDRP3 were successfully constructed and its growth and development were normal . The ΔAgo1 and ΔAgo2 strains reduced the stress effects on cell wall and osmotic pressure compared to the control . Ago1 gene deletion reduced the effect of H2 O2 , and conversely RDRP3 gene deletion increased the inhibition of H2 O2 . The Ago2 and RDRP1 strains reduced the effect on genetic damage agent . In addition , ΔRDRP1 increased the effect of osmotic stress . Conclusion The Ago1 , Ago2 , RDRP1 and RDRP3 genes of A. flavus are not in⁃ volved in the regulation of growth rate and asexual reproduction and can participate in the regulating of the host stress response to the environment .

For the perioperation of chronic rhinosinusitis (CRS), the local corticosteroid nasal spray, hormone and antibiotics oral treatment are mainly used in modern medicine. Oral treatment decoction, nasal spray and traditional Chinese medicine lavage and so on which are the combinations of internal and external treatment are used in TCM therapy. This article reviewed the use of integrated traditional Chinese and Western medicine in the perioperation of CRS, and provided references for standardization of integrated traditional Chinese medicine and Western medicine therapy for treating CRS.