OBJECTIVE: To explore the clinical phenotypes and genetic characteristics of five children with Lamb-Shaffer syndrome (LAMSHF). METHODS: Five children with LAMSHF diagnosed at the Department of Pediatrics, the First Medical Center of Chinese PLA General Hospital from April 2021 to December 2024 were selected as study subjects. Clinical data of the children was collected. Genomic DNA was extracted from peripheral blood samples of the children and their parents. Whole exome sequencing (WES) was carried out to screen for variants. This study was approved by the Medical Ethics Committee of the Chinese PLA General Hospital (Ethics No.: S2025-411-01). RESULTS: All five children had presented with global developmental delay. Among them, two had manifestations of autism spectrum disorder, two had abnormal electroencephalogram findings, four had abnormal MRI results, and two had ocular abnormalities. WES has detected five novel variants in the SOX5 gene. Among these, c.1771G>C (p.Gly591Arg) was unreported previously. Sanger sequencing confirmed that none of the parents had carried the same variants, suggesting that they were all de novo variants. According to the guidelines from the American College of Medical Genetics and Genomics (ACMG), two nonsense variants and one missense variant were classified as pathogenic, whilst two missense variants were classified as likely pathogenic. CONCLUSION This study has clarified the correlation between the clinical phenotypes of five children with LAMSHF and variants of the SOX5 gene, which expanded the mutational spectrum of the SOX5 gene and provided a basis for the clinical diagnosis and genetic counseling.
Humans ; Male ; Female ; Phenotype ; Child, Preschool ; Child ; SOXD Transcription Factors/genetics* ; Exome Sequencing ; Mutation ; Infant

The aim of this study is to establish an gene editing method of Mesomycoplasma hyo-pneumoniae(Mhp)based on the homologous recombination principle.The restriction enzyme di-gestion and ligation method combined with gene synthesis were used to construct a shuttle plasmid to achieve replication in both Mhp and Escherichia coli(E.coli).The pGEM?-T vector was used as the skeleton.The oriC sequence of Mhp which can achieve the replication of the plasmid in Mhp was inserted into the vector.Sequences of the Spiroplasma promoter and puromycin resistance gene were then inserted into the above constructed plasmid to screen recombinant clones.The up-stream and downstream homologous arms of p97 were constructed to initiate homologous recombination.The recA gene of E.coli is inserted to improve the efficiency of homologous recom-bination.The obtained shuttle plasmid was then delivered into Mhp by electro-transformation or chemical transformation.A shuttle plasmid,pGEM?-Mhp-oriC-p 97,which can replicate in both Mhp and E.coli was constructed.With the transformation of this plasmid,the carried puromycin gene and recA gene can be expressed,the p97 gene can be edited.Finally,the genetically unstable p97 gene mutant was initially obtained.In this study,a tool for Mhp gene editing based on the principle of homologous recombination was established,which laid a foundation for the develop-ment of tools for studying the pathogenesis of Mhp.

The aim of this study is to establish an gene editing method of Mesomycoplasma hyo-pneumoniae(Mhp)based on the homologous recombination principle.The restriction enzyme di-gestion and ligation method combined with gene synthesis were used to construct a shuttle plasmid to achieve replication in both Mhp and Escherichia coli(E.coli).The pGEM?-T vector was used as the skeleton.The oriC sequence of Mhp which can achieve the replication of the plasmid in Mhp was inserted into the vector.Sequences of the Spiroplasma promoter and puromycin resistance gene were then inserted into the above constructed plasmid to screen recombinant clones.The up-stream and downstream homologous arms of p97 were constructed to initiate homologous recombination.The recA gene of E.coli is inserted to improve the efficiency of homologous recom-bination.The obtained shuttle plasmid was then delivered into Mhp by electro-transformation or chemical transformation.A shuttle plasmid,pGEM?-Mhp-oriC-p 97,which can replicate in both Mhp and E.coli was constructed.With the transformation of this plasmid,the carried puromycin gene and recA gene can be expressed,the p97 gene can be edited.Finally,the genetically unstable p97 gene mutant was initially obtained.In this study,a tool for Mhp gene editing based on the principle of homologous recombination was established,which laid a foundation for the develop-ment of tools for studying the pathogenesis of Mhp.

Objective To investigate the difference of spontaneous hemorrhagic transformation(HT)in enlarged perivascular space(EPVS)of different site and various dilation in patients with acute ischemic stroke.Methods A total of 595 patients admitted to our department acute ischemic stroke from January 2021 to June 2024 were recruited in this study.Brain magnetic resonance ima-ging was conducted in all of them within 72 h after admission,and their EPVS status was collect-ed.According to the results of brain MRI,they were divided into EPVS(384 cases)and non-EPVS(211 cases)groups.Based on EPVS site,the EPVS group was further assigned into basal ganglia(135 cases),centrum semiovale(132 cases),and midbrain(117 cases)subgroups,and based on the severity,the group was also into mild(155 cases)and severe(229 cases)subgroups.Brain CT or MRI examination was re-performed on the 7th to 10th day during hospitalization to investigate the intracranial condition,and then spontaneous HT was evaluated.The incidence of spontaneous HT in the EPVS patients of different sites and severity was analyzed and compared.Results The incidence of spontaneous HT was significantly higher in the EPVS group than the non-EPVS group(27.08％vs 8.53％,P＜0.01),and the incidence of spontaneous HT was also obviously higher in the basal ganglia subgroup than the centrum semiovale subgroup and then the midbrain subgroup(40.00％vs 21.97％vs 17.95％,P＜0.01).What's more,the severe subgroup had nota-bly higher incidence of spontaneous HT than the mild subgroup(33.19％vs 18.06％,P＜0.01).Conclusion There is difference in spontaneous HT among acute ischemic stroke patients with EPVS of different site and severity.EPVS in the basal ganglia and of severe level indicates a high-er risk of spontaneous HT,and the two features can better predict the incidence of spontaneous HT.

Objective To investigate the difference of spontaneous hemorrhagic transformation(HT)in enlarged perivascular space(EPVS)of different site and various dilation in patients with acute ischemic stroke.Methods A total of 595 patients admitted to our department acute ischemic stroke from January 2021 to June 2024 were recruited in this study.Brain magnetic resonance ima-ging was conducted in all of them within 72 h after admission,and their EPVS status was collect-ed.According to the results of brain MRI,they were divided into EPVS(384 cases)and non-EPVS(211 cases)groups.Based on EPVS site,the EPVS group was further assigned into basal ganglia(135 cases),centrum semiovale(132 cases),and midbrain(117 cases)subgroups,and based on the severity,the group was also into mild(155 cases)and severe(229 cases)subgroups.Brain CT or MRI examination was re-performed on the 7th to 10th day during hospitalization to investigate the intracranial condition,and then spontaneous HT was evaluated.The incidence of spontaneous HT in the EPVS patients of different sites and severity was analyzed and compared.Results The incidence of spontaneous HT was significantly higher in the EPVS group than the non-EPVS group(27.08％vs 8.53％,P＜0.01),and the incidence of spontaneous HT was also obviously higher in the basal ganglia subgroup than the centrum semiovale subgroup and then the midbrain subgroup(40.00％vs 21.97％vs 17.95％,P＜0.01).What's more,the severe subgroup had nota-bly higher incidence of spontaneous HT than the mild subgroup(33.19％vs 18.06％,P＜0.01).Conclusion There is difference in spontaneous HT among acute ischemic stroke patients with EPVS of different site and severity.EPVS in the basal ganglia and of severe level indicates a high-er risk of spontaneous HT,and the two features can better predict the incidence of spontaneous HT.

Objective:To explore the characteristic of dynamic function network connectivity (dFNC) of resting brain networks in internet gaming disorder (IGD) adolescents.Methods:Forty-four adolescent IGD subjects (IGD group, male/female: 38/6) and fifty healthy controls (HC group, male/female: 40/10) were collected, and the subjects completed demographic questionnaires, Young internet addiction scale(YIAS), Chinese adolescents' maladaptive cognitions scale(CAMAS), and functional magnetic resonance imaging (fMRI) tests. The fMRI data were preprocessed on the Matlab platform, and the preprocessed data was divided into 64 components for group level independent component analysis.The dynamic functional connectivity of obtained 18 effective independent components was analyzed by sliding time window technique, and the difference of dynamic functional connectivity of brain triple network between the IGD group and HC group was compared using SPSS 22.0 software.Results:Four repeated dFNC states were identified through cluster analysis.Each state indicated that different functional networks had different connection strengths.State 3, the most frequent state, had been indicated that the whole brain network of the subject was in a state of weak functional connectivity.The second frequent state was state 1, which indicated enhanced functional connectivity within the subject's central executive network (CEN).State 2 had been indicated enhanced functional connectivity within the subject's salience network (SN).State 4 had been indicated generally enhanced functional connectivity in the subjects' brain networks, and this state was the least frequent.The results of non-parametric permutation test on the time attribute showed that compared with the HC group, the IGD group had a longer time score (IGD group: 0.24±0.19, HC group: 0.13±0.15, t=1.19, P<0.05, non-parametric substitution test) for state 1 with strong connectivity within the CEN, which was positively correlated with the YIAS score and game time ( r=0.418, P=0.003; r=0.515, P=0.004).Compared with HC group, the functional connectivity of ICD group between the internal insula of the SN and the dorsal anterior cingulate cortex was enhanced ( P<0.05, FDR corrected), while the average residence time in weakly connected state 3 was longer ( Z=2.09, P<0.05, nonparametric substitution test). Conclusions:The difference in dynamic functional connectivity of the triple network in the brain of IGD adolescents under resting state is mainly manifested by strong connections in CEN, functional connections between insula and dorsal anterior cingulate cortex in SN is enhanced, and weakening of overall functional connections, which may play an important role in the pathological mechanism of IGD.

Objective To explore the influencing factors of device-related pressure injuries (DRPI) in critically ill patients, construct a risk prediction model, and validate its effectiveness. Methods A retrospective analysis was conducted on the clinical data of 136 critically ill patients. Based on the occurrence of DRPIs, the patients were divided into occurrence group (32 patients) and non-occurrence group (104 patients). Univariate analysis and binary Logistic regression analysis were used to investigate the influencing factors of DRPIs, and a binary Logistic regression model was constructed. The Hosmer-Lemeshow test was used to assess the goodness of fit of the model, and the area (AUC) under the receiver operating characteristic (ROC) curve was used to evaluate the predictive performance of the model.Additionally, the Bootstrap method was employed for internal validation of the model. Results Univariate analysis revealed statistically significant differences between the two groups in terms of age, diabetes, ICU stay duration, duration of non-invasive ventilation mask use, Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) score, Braden scale score, use of vasoactive drugs, prone position ventilation, hemoglobin levels, and lactate levels (P < 0.05). Binary Logistic regression analysis indicated that age, diabetes, ICU stay duration, APACHE Ⅱ score, duration of non-invasive ventilation mask use, use of vasoactive drugs, prone position ventilation, lactate levels, Braden scale score, and hemoglobin levels were independent influencing factors for DRPIs in critically ill patients (P < 0.05). A binary Logistic regression model for predicting the risk of DRPIs in critically ill patients was constructed based on these independent factors. The overall prediction accuracy of the model was 99.26%. The Hosmer-Lemeshow goodness-of-fit test showed that the model had good fitness (χ² < 0.001, P>0.999). Internal validation using the Bootstrap method and ROC curve analysis showed that the AUC of the model for predicting DRPIs in critically ill patientswas 0.952, with a sensitivity of 87.5% and a specificity of 93.3%. Conclusion The risk prediction model for DRPIs in critically ill patients constructed in this study demonstrates good stability and predictive performance. It can assist clinical healthcare professionals in screening high-risk patients and formulating personalized intervention plans.

Objective:To analyze the mediating role of maladaptive cognition between family relationships and internet gaming disorder (IGD) in adolescents.Methods:From September 2021 to April 2023, a total of 52 adolescents with IGD (IGD group) from the First Affiliated Hospital of Zhengzhou University and 51 normal adolescents (HC group) from community were selected as study subjects.The severity of adolescent internet addiction was assessed using the Young internet addiction scale (YIAS).Family relationships and maladaptive cognition were evaluated by the Chinese version of family environment scale (FES-CV) and the Chinese adolescents’ maladaptive cognitions scale (CAMCS).SPSS 22.0 software was used for descriptive analysis, correlation analysis and mediation analysis.Two-sample t-tests were used for quantitative data comparison, and chi-square tests were used for qualitative data comparison. Pearson correlation analysis was conducted to analyze the relationship of variables, and the intermediate variable test was performed using Bootstrap method. Results:(1) Compared with HC group, the score of YIAS in IGD group was higher, and the difference was statistically significant((63.69±10.36), (28.96±5.20), t=21.571, P<0.05). Compared with the HC group, the conflict score of FES-CV in the IGD group was higher((4.79±2.02), (3.33±1.42), t=4.228, P<0.05), and the cohesion score of FES-CV was lower ((4.46±1.64), (5.57±1.33), t=-3.768, P<0.05).Compared with the HC group, the CAMCS score in IGD group was higher, and the difference was statistically significant((28.88±10.11), (22.98±9.11), t=3.116, P<0.05). (2) There was a positive correlation between conflict of family relationships and IGD ( r=0.504, P<0.01), and a negative correlation between cohesion of family relationships and IGD( r=-0.474, P<0.01) in IGD group. (3) Path analysis and mediation effect test showed that conflict in family relationships positively predicted IGD ( β=0.331, 95% CI=0.100-0.563) and maladaptive cognition ( β=0.372, 95% CI=0.108-0.636). Maladaptive cognitive positively predicted IGD ( β=0.464, 95% CI=0.232-0.696). Maladaptive cognition played a mediating role between conflict of family relationships and IGD, and the mediating effect value was 0.173 (95% CI=0.020-0.366), accounting for 34.33%(0.173/0.504) of the total effect. Conclusion:Family relationships are related to IGD in adolescents. Maladaptive cognition may play a mediating role between family relationships and IGD in adolescents.

Objective:To analyze the mediating role of maladaptive cognition between family relationships and internet gaming disorder (IGD) in adolescents.Methods:From September 2021 to April 2023, a total of 52 adolescents with IGD (IGD group) from the First Affiliated Hospital of Zhengzhou University and 51 normal adolescents (HC group) from community were selected as study subjects.The severity of adolescent internet addiction was assessed using the Young internet addiction scale (YIAS).Family relationships and maladaptive cognition were evaluated by the Chinese version of family environment scale (FES-CV) and the Chinese adolescents’ maladaptive cognitions scale (CAMCS).SPSS 22.0 software was used for descriptive analysis, correlation analysis and mediation analysis.Two-sample t-tests were used for quantitative data comparison, and chi-square tests were used for qualitative data comparison. Pearson correlation analysis was conducted to analyze the relationship of variables, and the intermediate variable test was performed using Bootstrap method. Results:(1) Compared with HC group, the score of YIAS in IGD group was higher, and the difference was statistically significant((63.69±10.36), (28.96±5.20), t=21.571, P<0.05). Compared with the HC group, the conflict score of FES-CV in the IGD group was higher((4.79±2.02), (3.33±1.42), t=4.228, P<0.05), and the cohesion score of FES-CV was lower ((4.46±1.64), (5.57±1.33), t=-3.768, P<0.05).Compared with the HC group, the CAMCS score in IGD group was higher, and the difference was statistically significant((28.88±10.11), (22.98±9.11), t=3.116, P<0.05). (2) There was a positive correlation between conflict of family relationships and IGD ( r=0.504, P<0.01), and a negative correlation between cohesion of family relationships and IGD( r=-0.474, P<0.01) in IGD group. (3) Path analysis and mediation effect test showed that conflict in family relationships positively predicted IGD ( β=0.331, 95% CI=0.100-0.563) and maladaptive cognition ( β=0.372, 95% CI=0.108-0.636). Maladaptive cognitive positively predicted IGD ( β=0.464, 95% CI=0.232-0.696). Maladaptive cognition played a mediating role between conflict of family relationships and IGD, and the mediating effect value was 0.173 (95% CI=0.020-0.366), accounting for 34.33%(0.173/0.504) of the total effect. Conclusion:Family relationships are related to IGD in adolescents. Maladaptive cognition may play a mediating role between family relationships and IGD in adolescents.

OBJECTIVE To prepare spider fibroin membrane loaded with Periplaneta americana extract， and investigate its characterization， in vitro drug release property and cytotoxicity. METHODS Using natural spider silk collected from Chilobrachys guangxiensis as raw material， P. americana extract as model drug， the drug-loaded spider fibroin membrane （hereinafter referred to as drug-loaded membrane） was prepared by solvent casting method. The material matrix spider fibroin membrane without P. americana extract （hereinafter referred to as blank membrane） was prepared with same method. The membrane structure was characterized by static water contact angle， Fourier infrared chromatography， X-ray diffraction and scanning electron microscopy from different angles； drug release characteristics in artificial saliva were simulated in vitro to evaluate the drug sustained-release performance. MTT assay was adopted to validate the cytotoxicity of drug-loaded membrane. RESULTS The drug-loaded membrane was prepared， and the static water contact angle was less than 90°， which was less than that of blank membrane. The drug-loaded membrane showed the characteristic absorption peak to polypeptide of P. americana extract at 1 500-1 700 cm－1. X-ray diffraction and scanning electron microscopy also proved that the drug was successfully loaded into the pellicle. The release time of the pellicle in artificial saliva was more than 200 min. The MTT test results showed that the cell proliferation rates of blank membrane and drug-loaded membrane were 84.6% and 79.4% （both greater than 70%）， respectively， without significant potential cytotoxicity. CONCLUSIONS Drug-loaded membrane prepared with natural spider silk has a certain sustained-release effect in artificial saliva， which can be further developed as a drug sustained-release carrier with excellent biological characteristics and biocompatibility.