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 investigate the clinical characteristics and genetic etiology of a child with Cortical dysplasia, complex, with other brain malformations 4 (CDCBM4) and epilepsy due to a TUBG1 gene variant. Methods:A child diagnosed with CDCBM4 and epilepsy at the Children′s Medical Center of the Affiliated Hospital of Guangdong Medical University in May 2024 was selected as the study subject. Clinical data were retrospectively analyzed. Peripheral venous blood samples were collected from the child and her parents for genomic DNA extraction. Trio-based whole-exome sequencing (WES) was performed, and candidate variants were validated by Sanger sequencing. According to the Standards and Guidelines for the Interpretation of Sequence Variants established by the American College of Medical Genetics and Genomics (ACMG), candidate variants were classified for pathogenicity. This study was approved by the Medical Ethics Committee of the Affiliated Hospital of Guangdong Medical University (Ethics No.: PJ2021-097).Results:The child, a 4-month-old female infant, had no special facial features, normal limb muscle strength, and increased muscle tone of infantile onset, with generalized tonic-clonic seizures as the main manifestation. During seizures, she exhibited head retroflexion, tightly closed eyes, and tonic convulsions of the limbs, occurring approximately 2-3 times per day. Electroencephalogram suggested bilateral anterior predominant medium-to-high amplitude 7-8 Hz mixed rhythm discharges. Head MRI revealed ventricular system dilatation and pachygyria. Trio-WES results indicated that the child has harbored a TUBG1 gene variant of c. 776C>T (p.Ser259Leu). Sanger sequencing verification showed that neither of her parents had carried the same variant, confirming it as de novo in origin. According to the ACMG guidelines, the variant was rated as pathogenic (PS2+ PS3+ PM2_Supporting+ PP3). Combining the child′s clinical phenotype, the child was diagnosed as CDCBM4 with epilepsy. Conclusion:Children with CDCBM4 and epilepsy due to TUBG1 gene variants may show pachygyria or agyria and commonly present with intellectual and motor developmental delays and seizure disorders of variable severity. The heterozygous TUBG1 c. 776C>T (p.Ser259Leu) variant is likely the genetic etiology underlying this disorder. The results of this study has expanded the mutational spectrum of the TUBG1 gene associated with CDCBM4 and epilepsy.

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 the therapeutic effect of extracellular vesicles(EVs) of Aspergillus flavus(A. flavus) on A. flavus infection. Methods: The EVs of A. flavus were isolated using ultracentrifugation and detected/identified by nanoparticle tracking analysis(NTA). Quantitative real-time PCR(qRT-PCR) and enzyme-linked immunosorbent assay(ELISA) kits were used to assess the effect of A. flavus EVs on the polarization of bone marrowderived macrophages(BMDMs) and the expression levels of several cytokines,including tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ),interleukin-6(IL-6),and interleukin-10(IL-10). The Galleria mellonella(G.mellonella) infection model was constructed. Three concentration groups of A. flavus EVs(0. 2,2,and 20 μg/larvae) were set as the experimental groups,and PBS was used as the control group for the infection model.Meanwhile,three forms of negative control groups were established,including the PBS group,the pierced controlgroup, and the negative control group. The therapeutic effect of A. flavus EVs on A. flavus infection was evaluated by the survival rate of the G. mellonella infection models. Results: The particle size of A. flavus EVs ranged from 20 to550 nm. A. flavus EVs could polarize BMDMs into both M1 and M2 phenotypes and induce the production of cytokines,including TNF-α,IFN-γ,IL-6,and IL-10. The results of the G. mellonella infection model showed that A.flavus EVs could improve the survival rate of G. mellonella after A. flavus infection. Conclusion The EVs produced by A. flavus can promote the expression of both pro-inflammatory and anti-inflammatory cytokines in BMDMs,induce M1 polarization and M2 polarization of BMDMs,and increase the survival rate of G. mellonella after A. flavus infection.

OBJECTIVE: To investigate the clinical characteristics and genetic etiology of a child with Cortical dysplasia, complex, with other brain malformations 4 (CDCBM4) and epilepsy due to a TUBG1 gene variant. METHODS: A child diagnosed with CDCBM4 and epilepsy at the Children's Medical Center of the Affiliated Hospital of Guangdong Medical University in May 2024 was selected as the study subject. Clinical data were retrospectively analyzed. Peripheral venous blood samples were collected from the child and her parents for genomic DNA extraction. Trio-based whole-exome sequencing (WES) was performed, and candidate variants were validated by Sanger sequencing. According to the Standards and Guidelines for the Interpretation of Sequence Variants established by the American College of Medical Genetics and Genomics (ACMG), candidate variants were classified for pathogenicity. This study was approved by the Medical Ethics Committee of the Affiliated Hospital of Guangdong Medical University (Ethics No.: PJ2021-097). RESULTS: The child, a 4-month-old female infant, had no special facial features, normal limb muscle strength, and increased muscle tone of infantile onset, with generalized tonic-clonic seizures as the main manifestation. During seizures, she exhibited head retroflexion, tightly closed eyes, and tonic convulsions of the limbs, occurring approximately 2-3 times per day. Electroencephalogram suggested bilateral anterior predominant medium-to-high amplitude 7-8 Hz mixed rhythm discharges. Head MRI revealed ventricular system dilatation and pachygyria. Trio-WES results indicated that the child has harbored a TUBG1 gene variant of c.776C>T (p.Ser259Leu). Sanger sequencing verification showed that neither of her parents had carried the same variant, confirming it as de novo in origin. According to the ACMG guidelines, the variant was rated as pathogenic (PS2+PS3+PM2_Supporting+PP3). Combining the child's clinical phenotype, the child was diagnosed as CDCBM4 with epilepsy. CONCLUSION Children with CDCBM4 and epilepsy due to TUBG1 gene variants may show pachygyria or agyria and commonly present with intellectual and motor developmental delays and seizure disorders of variable severity. The heterozygous TUBG1 c.776C>T (p.Ser259Leu) variant is likely the genetic etiology underlying this disorder. The results of this study has expanded the mutational spectrum of the TUBG1 gene associated with CDCBM4 and epilepsy.
Humans ; Female ; Epilepsy/genetics* ; Malformations of Cortical Development/genetics* ; Infant ; Phenotype ; Exome Sequencing ; Microtubule-Associated Proteins/genetics*

OBJECTIVE: To explore the clinical phenotype and genetic etiology of a child with Ehlers-Danlos syndrome, spondylodysplastic type 2 (EDSSPD2). METHODS: A child who was admitted to the Children's Medical Center of the Affiliated Hospital of Guangdong Medical University in July 2024 for "delayed motor development for 1 and a half year" was selected as the study subject. Clinical data of the child was collected, including medical history, family history, and results of auxiliary examinations. Peripheral venous blood samples were collected from the child and his two brothers and both parents. Genomic DNA was extracted from the child and his family members and subjected to whole-exome sequencing (WES) and copy number variation (CNV) analysis. Sanger sequencing was used to verify the parental origin of the candidate variants. Multiple protein function prediction software tools, including SIFT, PolyPhen-2, and REVEL, were used to assess the impact of candidate variants on the protein function. Based on protein database information from UniProt, a two dimensional structural schematic of the target protein was generated. The pathogenicity of the variants was classified based on the guidelines from the American College of Medical Genetics and Genomics (ACMG). Relevant literature on the B3GALT6 gene variants leading to EDSSPD2 was retrieved from CNKI, Wanfang Data Knowledge Service Platform, and PubMed databases. The procedures followed in this study were reviewed and approved by the Medical Ethics Committee of Affiliated Hospital of Guangdong Medical University (Ethics No.：PJ2021-097). RESULTS: The proband was a 2-year-old male with an onset in infancy. The main clinical manifestations included loose skin, scoliosis and kyphosis, generalized hypermobility of joints, and motor developmental delay. WES has revealed two compound heterozygous variants of the B3GALT6 gene (NM_080605.4): c.766C>T (p.Arg256Trp) and c.962G>A (p.Cys321Tyr). Sanger sequencing verification showed that the c.766C>T and c.962G>A variants were respectively derived from his phenotypically normal father and mother. Bioinformatics analysis showed that for the c.766C>T (p.Arg256Trp) variant, the Arg256 site is located within the galactosyltransferase catalytic domain (GalT domain) of the β3GalT6 protein. According to the ACMG guidelines, the c.766C>T variant was classified as a likely pathogenic (PS3+PM2_supporting+PM3+PP3), and the c.962G>A was classified as a variant of unknown significance (PM2_Supporting+PM3+PP3). By following the pre-set literature retrieval strategy, a total of 12 articles related to B3GALT6 gene variants were identified (11 English and 1 Chinese), which involved a total of 71 patients. Among these, 4 reports (involving 20 patients) involved B3GALT6 gene variants leading to EDSSPD2. Among the 18 live-born EDSSPD2 patients (including the proband in this study), common clinical manifestations have included scoliosis (88.9%, 16/18), generalized hypotonia (83.3%, 15/18), and soft and lax skin (66.7%, 12/18). Some patients already showed skeletal abnormalities on prenatal ultrasound scan (22.2%, 4/18), while a few presented with cervical instability (16.7%, 3/18). One child had deceased at 18 months of age due to hypoxia caused by tracheomalacia and tracheal compression due to scoliosis. Among the 23 reported EDSSPD2 related B3GALT6 variant sites, missense variants were the most common (78.3%, 18/23), followed by nonsense variants (21.7%, 5/23). CONCLUSION Above finding has enriched the clinical and mutational spectra of EDSSPD2. Early genetic testing has important clinical value for the diagnosis, differential diagnosis, and genetic counseling of this disease.
Humans ; Male ; Ehlers-Danlos Syndrome/genetics* ; Pedigree ; N-Acetylgalactosaminyltransferases/genetics* ; Asian People/genetics* ; DNA Copy Number Variations ; Exome Sequencing ; Female ; Child ; Child, Preschool ; Phenotype ; Mutation ; China ; East Asian People ; Galactosyltransferases

Objective:To investigate the clinical characteristics and genetic etiology of a child with Cortical dysplasia, complex, with other brain malformations 4 (CDCBM4) and epilepsy due to a TUBG1 gene variant. Methods:A child diagnosed with CDCBM4 and epilepsy at the Children′s Medical Center of the Affiliated Hospital of Guangdong Medical University in May 2024 was selected as the study subject. Clinical data were retrospectively analyzed. Peripheral venous blood samples were collected from the child and her parents for genomic DNA extraction. Trio-based whole-exome sequencing (WES) was performed, and candidate variants were validated by Sanger sequencing. According to the Standards and Guidelines for the Interpretation of Sequence Variants established by the American College of Medical Genetics and Genomics (ACMG), candidate variants were classified for pathogenicity. This study was approved by the Medical Ethics Committee of the Affiliated Hospital of Guangdong Medical University (Ethics No.: PJ2021-097).Results:The child, a 4-month-old female infant, had no special facial features, normal limb muscle strength, and increased muscle tone of infantile onset, with generalized tonic-clonic seizures as the main manifestation. During seizures, she exhibited head retroflexion, tightly closed eyes, and tonic convulsions of the limbs, occurring approximately 2-3 times per day. Electroencephalogram suggested bilateral anterior predominant medium-to-high amplitude 7-8 Hz mixed rhythm discharges. Head MRI revealed ventricular system dilatation and pachygyria. Trio-WES results indicated that the child has harbored a TUBG1 gene variant of c. 776C>T (p.Ser259Leu). Sanger sequencing verification showed that neither of her parents had carried the same variant, confirming it as de novo in origin. According to the ACMG guidelines, the variant was rated as pathogenic (PS2+ PS3+ PM2_Supporting+ PP3). Combining the child′s clinical phenotype, the child was diagnosed as CDCBM4 with epilepsy. Conclusion:Children with CDCBM4 and epilepsy due to TUBG1 gene variants may show pachygyria or agyria and commonly present with intellectual and motor developmental delays and seizure disorders of variable severity. The heterozygous TUBG1 c. 776C>T (p.Ser259Leu) variant is likely the genetic etiology underlying this disorder. The results of this study has expanded the mutational spectrum of the TUBG1 gene associated with CDCBM4 and epilepsy.

Objective To investigate the correlation between the gene polymorphisms of rs7100927,rs10885409,rs4918789,rs290487 of TCF7L2 gene and type 2 diabetes mellitus(T2DM)of Uygur nationality and Han nationality.Methods 577 T2DM patients(T2DM group,293 Han and 284 Uyghu)were enrolled in the Department of Endocrinology,Affiliated Hospital of Traditional Chinese Medicine of Xinjiang Medical University from December 2013 to August 2015.In the same period,307 healthy subjects were selected as normal control(NC)group,of which 208 were Han Chinese and 99 were Uyghu.The polymorphism of TCF7L2 gene rs7100927,rs10885409,rs4918789 and rs290487 were identified by imLDRTM multiple SNP typing technique.Results In Uygur,the frequency of allele G at rs7100927 and rs4918789 of TCF7L2 gene and the frequency of allele C at rs10885409 and rs290487 in T2DM group were lower than those in NC group(P<0.05).Conclusion The G allele at rs7100927 and rs4918789 and C allele at rs10885409 and rs290487 of TCF7L2 gene may be protective factors for T2DM in Xinjiang Uygur population.

Objective To compare the effect of glycopyrrolate or atropine in combination with neostigmine on adverse cardiovascular events(ACEs)after operation in elderly patients undergoing laparo-scopic surgery.Methods A total of 142 patients scheduled for elective laparoscopic surgery were enrolled,69 males and 73 females,aged 65-80 years,BMI 18-28 kg/m2,ASA physical status Ⅰ or Ⅱ.The pa-tients were randomly divided into two groups:the glycopyrrolate group(group G)and the atropine group(group A),71 patients in each group.After the last administration of muscle relaxants for more than 30 mi-nutes,antagonizing residual neuromuscular blockade was performed.Glycopyrrolate 4 μg/kg and neostigmine 20 μg/kg were given intravenously in group G,atropine 10 μg/kg and neostigmine 20 μg/kg were given intravenously in group A.The incidence of ACEs and severe ACEs during operation and 72 hours after operation were recorded.Recovery situation in PACU such as NRS scores at rest and coughing,Rich-mond agitation-sedation scale(RASS)score,and modified Aldrete score 15 and 30 minutes after extubation were recorded.Emergence agitation,dry mouth,nausea,vomiting,and delirium 24 hours after operation were recorded.Results Compared with group A,the total incidence of ACEs,tachycardia,and myocardial ischemia after operation were significantly decreased in group G(P＜0.05),the incidence of dry mouth 24 hours postoperatively was significantly increased in group G(P＜0.05).There was no severe ACEs oc-curred in the two groups 72 hours after operation.Conclusion Compared with atropine,glycopyrrolate combined with neostigmine in elderly patients undergoing laparoscopic surgery can reduce the incidence of cardiac tachycardia,myocardial ischemia,and total ACEs after operation,and there was no severe ACEs occurred.However,it can increase the incidence of dry mouth 24 hours after operation.

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.