Objective:To investigate the clinical and genetic characteristics of KDM6B gene variation associated neurological developmental disorder in a child. Methods:Clinical data were collected from a child of KDM6B gene variation associated neurological developmental disorder admitted to Children′s Hospital Affiliated to Zhengzhou University in July 2021. His clinical manifestations and genetic variation profiles were retrospectively analyzed and literature review was conducted. Results:The patient was a one-year-six-month old male, with protruding forehead, joint laxity, distal skeletal abnormalities, and behavioral, cognitive, language, intellectual, and psychomotor development disorder. The whole-exome sequencing and Sanger sequencing confirmed that there was a de novo heterozygous frameshift variation c.1718delC(p.Pro573Hisfs *9) in exon 11 of the KDM6B gene. This variation was classified as pathogenic (PVS1+PS2+PM2_supporting) according to the American College of Medical Genetics and Genomics and the Association for Molecular Pathology guidelines, with no prior reports. By literature review, no relevant Chinese literature was retrieved, whereas 4 English literatures were found, reporting 98 patients, totally 99 patients (including this case) with nervous system development disorder due to KDM6B gene variation. The main manifestations were neurodevelopmental disorders such as speech, motor, and behavioral abnormalities, mental retardation, as well as facial deformities, hypotonia, infantile feeding difficulties/gastroesophageal reflux, joint/ligament laxity, and abnormalities of the hands and toes/palms. A total of 83 variation sites were found, including 37 frameshift variations, 18 missense variations, 21 nonsense variations, and 7 splicing variations, all of which were heterozygous variations. Conclusions:The KDM6B gene variation can lead to neurodevelopmental disorder, craniofacial developmental and skeletal abnormalities. The de novo heterozygous variation in the KDM6B gene is considered to be the genetic etiology of this child. This study extends the spectrum of KDM6B gene variant.

Objective:To investigate the clinical and genetic characteristics of KDM6B gene variation associated neurological developmental disorder in a child. Methods:Clinical data were collected from a child of KDM6B gene variation associated neurological developmental disorder admitted to Children′s Hospital Affiliated to Zhengzhou University in July 2021. His clinical manifestations and genetic variation profiles were retrospectively analyzed and literature review was conducted. Results:The patient was a one-year-six-month old male, with protruding forehead, joint laxity, distal skeletal abnormalities, and behavioral, cognitive, language, intellectual, and psychomotor development disorder. The whole-exome sequencing and Sanger sequencing confirmed that there was a de novo heterozygous frameshift variation c.1718delC(p.Pro573Hisfs *9) in exon 11 of the KDM6B gene. This variation was classified as pathogenic (PVS1+PS2+PM2_supporting) according to the American College of Medical Genetics and Genomics and the Association for Molecular Pathology guidelines, with no prior reports. By literature review, no relevant Chinese literature was retrieved, whereas 4 English literatures were found, reporting 98 patients, totally 99 patients (including this case) with nervous system development disorder due to KDM6B gene variation. The main manifestations were neurodevelopmental disorders such as speech, motor, and behavioral abnormalities, mental retardation, as well as facial deformities, hypotonia, infantile feeding difficulties/gastroesophageal reflux, joint/ligament laxity, and abnormalities of the hands and toes/palms. A total of 83 variation sites were found, including 37 frameshift variations, 18 missense variations, 21 nonsense variations, and 7 splicing variations, all of which were heterozygous variations. Conclusions:The KDM6B gene variation can lead to neurodevelopmental disorder, craniofacial developmental and skeletal abnormalities. The de novo heterozygous variation in the KDM6B gene is considered to be the genetic etiology of this child. This study extends the spectrum of KDM6B gene variant.

Objective To observe the value of cardiac CT angiography(CCTA)for left atrial appendage closure(LAAC).Methods Twelve Labradors were enrolled and underwent LAAC.CCTA examinations were performed before and 7,45 and 90 days after LAAC,and the left atrial appendage(LAA)parameters before and post LAAC occluder compression ratio were recorded.Ninety days after LAAC,the dogs were euthanized for gross anatomical observation,micro CT imaging and pathological examination,and the value of CCTA for LAAC was analyzed.Results Based on CCTA before LAAC,LAA opening depth was(18.8±2.6)mm,the minimum diameter was(16.3±2.0)mm and the maximum diameter was(22.6±3.2)mm,so 28 mm diameter occluders were selected for LAAC.Seven,45 and 90 days after LAAC,no displacement of occluder,thrombi nor device leakage was found on CCTA,while no significant difference of compression ratio was found among different time points(P＞0.05),nor between those measured with CCTA and micro CT 90 days after LAAC(P＞0.05).Ninety days after LAAC,gross dissection revealed complete endothelialization of occlude in 11 dogs and incomplete endothelialization in 1 dog.Pathological examination showed that collagen fibers arranged neatly,with endothelial cells covering the surface,and no inflammatory cell infiltration was observed.Conclusion CCTA could be used to observe LAA and surrounding structures before LAAC,hence guiding to select appropriate occluder,evaluate occlusion effect and monitor complications of LAAC in dogs.

Objective: To investigate the correlation between campus bullying and suicidal tendency symptoms comorbidity with addictive behavior among middle and high school students in Hainan Province, so as to provide a theoretical basis for health education and behavioral intervention in schools. Methods: In July 2023, an anonymous questionnaire survey was conducted among 6 654 middle and high school students in Hainan Province, selected by probability proportional sampling and stratified cluster random sampling method. Campus bullying, suicidal tendency and addictive behavior were determined according to the relevant items in the questionnaire on health related behaviors of Chinese students health status and influencing factors questionnaire, and self designed questionnaire. The co occurrence of campus bullying and suicidal tendency among students was analyzed. The binary Logistic regression method was used to analyze the correlation between the co occurrence of campus bullying and suicidal tendency and the addictive behavior of middle school students. Results: The report rate of campus bullying among middle and high school students in Hainan Province was 28.48%, the suicidal tendency was 15.25%, and the co occurrence of campus bullying and suicidal tendency was 8.00%. The results of Logistic regression analysis showed that middle school students and left behind students were prone to campus bullying and suicide tendency ( OR =1.55, 1.52, P <0.05), while Internet addiction, gambling and current smoking showed significant positive correlation with comorbidity of campus bullying and suicide tendency ( OR =3.14, 2.18, 2.07, P < 0.05 ). Conclusions Middle and high school students with addictive behavior have a higher possibility of comorbidity of campus bullying and suicidal tendency. The comprehensive intervention of addictive behavior can reduce the incidence of co occurrence of campus bullying and suicidal tendency, so as to improve health and wellbeing of middle school students.

Objective This study aimed to estimate the therapeutic effects of preformed metal crown(PMC)and pre-fabricated zirconia crowns(ZC)on decayed primary morals in children,as well as to analyze the possible influencing fac-tors.Methods A retrospective cohort study was performed on the data of 192 patients(aged 3 to 8)in the Stomatologi-cal Department of Shenzhen Children's Hospital from October 2021 to December 2021.The decayed mandibular first molars were selected and restored by vital-pulp therapy followed by PMC and ZC,including 96 cases(96 caries)in the PMC group and 96 cases(96 caries)in the ZC group.Oral clinical examination was performed at 3 months,1 year,and 2 years after treatment,overwiewing the clinical therapeu-tic effects and periodontal status of PMC and ZC groups,as well as recording the crown integrity,gingival index(GI),probing bleeding index(BI),plaque index(PLI)and various prosthetic indices.Results No significance differences existed in the periodontal status of PMC and ZC groups at 3 months,1 year,and 2 years after treatment(P>0.05).However,the GI,BI,and PLI in the PMC group were higher than those in the ZC group at 3 months,1 year,and 2 years after treatment,and the difference was dramatically signifi-cant(P<0.05).No significances difference existed in various prosthetic indices(P>0.05),as well as in the GI,BI,and PLI,between the two groups(P>0.05).No significant differences existed in various prosthetic indices between genders after PMC restoration(P>0.05).The scores of girls in various prosthetic indices after ZC restoration were higher than those of boys(P<0.05).Pearson correlation analysis indicated an inverse correlation between age in the PMC group and the GI,BI,PLI,and FDI indices(P<0.01),rather than in the ZC group(P>0.05).Conclusion PMC and ZC can be ap-plied to restore deciduous molar caries.The periodontal status of deciduous teeth in ZC group was superior to that in the PMC group.The periodontal status of deciduous teeth in PMC group may be stable with increased age.

【Objective】 To analyze the current situation of human parvovirus B19 infection in blood donors in different regions of China, so as to provide basis for formulating reasonable screening programs of B19 virus for blood donors in various cities and regions. 【Methods】 The literatures related to human parvovirus B19 infection in whole blood and plasma donors published from 1998 to 2021 were searched in the database, and meta-analysis of literatures that satisfied the inclusion criteria was conducted by R4.1.0 software. 【Results】 A total of 35 literatures were obtained, 20 literatures involving 56 846 blood donor samples and 8 literatures involving 1 608 pooled raw plasma samples were subjected to Meta analysis of the positive rates of B19 DNA; 17 literatures involving 12 308 blood sample were subjected to the Meta analysis of the positive rate of B19 IgG antibody.The positive rates of B19 DNA in blood donors(I²=96%, τ²=0.026 0, P<0.01)and pooled raw plasma (I²=98%, τ²=0.124 5, P<0.01), as well as the positive rate of B19 IgG antibod (I²=98%, τ ² =0.021 0, P < 0.01) presented significant heterogeneity between regions. The combined positive rate of B19 DNA was estimated to be 2.0% (95%-CI: 0.007~0.039), that of pooled raw plasma for production was 66.6% (95%-CI: 0.476~0.832), and that of B19 IgG antibody was 30.2% (95%-CI: 0.246~0.357). 【Conclusion】 Low HPV B19 infection rate and high positive rate of IgG antibody were found in blood donors. Therefore, the risk of B19 virus infection due to blood transfusion is low, but B19 infections in blood donors varied significantly between regions. Domestic cities and regions should reasonably evaluate their own B19 virus infection status to formulate appropriate B19 virus screening programs for blood donors, so as to reduce blood transfusion transmitted risk of B19 virus. In addition, the infection rate of B19 virus in pooled plasma for production is somewhat high. Recipients should be screened for B19 virus antibodies, and appropriate blood transfusion schemes should be formulated for blood recipients lacking neutralizing B19 IgG antibodies to reduce the exposure of B19 virus.

Clinical data and genetic mutation characteristics of a patient with Coffin-Siris syndrome by 6q25.3 deletion were summarized. The child was a 7-year and 6-month old girl who had feeding difficulties, repeated infection, language and motor retardation, low intelligence, laryngeal cartilage dysplasia, thick eyebrows, sparse teeth, hairy back, hyperactivity and aggressive behavior, seizures and ataxia. There was no abnormality in chromosomal karyotype analysis by proband; genomic copy number variant sequencing (CNV-seq) indicated approximately 4.27 Mb heterozygous deletion in chromosome 6q25.3 region, with 17 genes including ARID1B gene, father maternal CNV-seq showing no abnormalities. Trio-whole-exome sequencing showed the proband missed all exons 1-20 of the ARID1B gene, with wild-type parents. The proband had severe clinical symptoms and haplodose insufficiency which was the genetic etiology.

Imbalance of calcium homeostasis and abnormal autophagy are important pathogenesis mechanisms for neurodegenerative diseases, as PD, AD, and amyotrophic lateral sclerosis (ALS); and correlation is noted between imbalance of calcium homeostasis and abnormal autophagy. A number of studies have reported that different calcium storage compartments of cells can affect autophagy via calcium channels or Ca 2+-related signal proteins, such as cytoplasm calcium release-activated calcium modulator 1 (Orai1) and transient receptor potential canonical channel (TRPC), endoplasmic reticulum calcium channel inositol 1,4,5-triphosphate receptor (IP3R), mitochondrial calcium uniporter (MCU) related to Ca 2+ uptake, lysosomal calcium channel transient receptor potential channel mucolipin 1(TRPML1) and two-pore channels, cytoplasmic Ca 2+/calmodulin-dependent kinase kinase β (CaMKKβ)/AMP-activated protein kinase(AMPK) pathway. This review summarizes the research progress on autophagy regulated by Ca 2+ in intracellular calcium storage compartments and cytoplasm in neurodegenerative diseases, with a view to further understand the pathogenesis of these diseases.

Objective:To explore the effects and mechanisms of polycaprolactone-cellulose acetate (PCL-CA) nanofiber scaffold loaded with rat epidermal stem cells (ESCs) on wound healing of full-thickness skin defects in rats.Methods:The experiment research method was applied. The primary ESCs were isolated from 1-3 d old Sprague-Dawley (SD) rats (undefined gender) by rapid adherent method and cultured by rapid adherent method. ESCs of the first passage were used for the subsequent experiments after the positive expressions of integrin β 1 and cytokeratin 19 (CK19) in primary cells were identified respectively by flow cytometey and immunofluorescence method. PCL-CA nanofiber scaffolds with polycaprolactone and cellulose acetate as components were prepared by electrospinning technique. The topological structure of the nanofiber scaffolds was determined and the diameter of 25 fibers was measured by scanning electron microscope. The constructed PCL-CA nanofiber scaffolds were used as the culture substrate for ESCs, which were cultured in keratinocytes (KCs) medium to construct ESCs-nanofiber scaffold complex (hereinafter referred to as ESCs scaffold). After 3 days of culture, the morphology of ESCs in the scaffold and their relationship was observed by scanning electron microscopy. The ESCs in ESCs scaffold were set as PCL-CA nanofiber scaffold group, and the ESCs cultured with KCs medium in culture dishes coated with type Ⅳ collagen were set as type Ⅳ collagen group. Western blotting was used to detect the protein expression level of CK19 in ESCs in the two groups after 3 days of culture ( n=3). The protein expressions of CK19 and proliferating nuclear antigen (PCNA) in ESCs in the two groups were detected by immunofluorescence method after 7 days of culture. A circular full-thickness skin wound of about 2 cm in diameter was prepared on both left and right sides of the back of 15 male SD rats aged 6-8 weeks. The rats were then equally divided into blank control group without implantation, scaffold alone group implanted with PCL-CA nanofiber scaffold, and ESCs scaffold group implanted with ESCs scaffold which were constructed after 3 days of culture according to the random number table. The percentage of wound areas on post injury day (PID) 3, 7, 14, and 21 was calculated ( n=5). The new skin tissue at the wound edge was collected on PID 21, the wound healing quality was evaluated by Masson staining, and the protein expression levels of Notch1, Jagged1, and Hes1, which are key proteins of Notch signaling pathway, were detected by Western blotting ( n=3). Data were statistically analyzed with one-way analysis of variance, one-way analysis of variance, analysis of variance for repeated measurement, independent sample t test, and Bonferroni correction. Results:The constructed PCL-CA nanofiber scaffolds had a porous, mesh-like, and multilayered three-dimensional structure, in which the surface of the fibers was smooth and non-porous, and the fiber diameter was (383±24) nm. The ESCs in ESCs scaffold showed intact cellular structures and were tightly attached to the scaffold after 3 days of culture. The cells were interconnected and fully extended on the surface of the scaffold to form a membrane. After 3 days of culture, the protein expression level of CK19 of ESCs in PCL-CA nanofiber scaffold group was significantly higher than that in type Ⅳ collagen group ( t=24.56, P<0.01). After 7 days of culture, compared with those in type Ⅳ collagen group, there was no significant change in the proportion of PCNA positive cells of ESCs in PCL-CA nanofiber scaffold group, while the proportion of CK19 positive cells was higher. On PID 3, 7, 14, and 21, the percentages of wound areas of rats in ESCs scaffold group were (78.0±1.8)%, (40.9±2.0)%, (17.9±1.1)%, and (5.0±1.0)%, respectively, which were significantly lower than (84.2±1.9)%, (45.4±2.6)%, (21.8±1.7)%, and (10.1±1.1)% in blank control group ( t=5.42, 3.09, 4.33, 7.58, P<0.05 or P<0.01) and (82.7±1.2)%, (44.8±2.0)%, (22.4±2.4)%, and (10.3±2.4)% in scaffold alone group ( t=4.98, 3.11, 3.84, 4.57, P<0.05 or P<0.01), while the percentages of wound areas of rats between blank control group and scaffold alone group were similar ( t=1.47, 0.39, 0.47, 0.22, P>0.05). On PID 21, the layer of new skin at the wound edge of rats in each group was intact; compared with that in blank control group or scaffold alone group, the new skin tissue at the wound edge of rats in ESCs scaffold group had more orderly collagen arrangement; the scaffolds in the new skin at the wound edge of rats were completely degraded in ESCs scaffold group and scaffold alone group. On PID 21, the protein expression levels of Notch1, Jagged1, and Hes1 in the new skin tissue at the wound edge of rats in scaffold alone group were similar to those in blank control group ( t=1.70, 1.94, 0.18, P>0.05), while the protein expression levels of Notch1, Jagged1, and Hes1 in the new skin tissue at the wound edge of rats in ESCs scaffold group were significantly higher than those in scaffold alone group ( t=13.31, 22.07, 20.71, P<0.01). Conclusions:PCL-CA nanofiber scaffolds can inhibit the differentiation of ESCs of rats without affecting their proliferation in vitro. ESCs scaffolds constructed through using PCL-CA nanofiber scaffolds as the carrier to culture ESCs of rats can significantly promote the wound healing of full-thickness skin defects in rats, and the mechanism may be related to the activation of Notch signaling pathway.

Objective:To disclose the genome characteristics and mutations of 2019 novel coronavirus (2019-nCoV) strains from the imported cases of Coronavirus Disease 2019 (COVID-19) in Gansu province, thereby to provide scientific reference for the prevention and control of COVID-19 in Gansu province. Methods:The respiratory tract specimens of imported COVID-19 cases from seven countries in Gansu province in 2020 were collected. The virus genome was sequenced by the second-generation sequencing technology, the whole genome sequences were compared and analyzed, and the MEGA software was used to construct a phylogenetic tree based on the neighbor-joining method.Results:A total of 46 2019-nCoV genome sequences with a length of 29 605~29 903 bp were obtained. Compared with the Wuhan reference strain (GenBank ID: NC_045512.2), it was found that the median (minimum to maximum) number of the nucleotide mutations of the 2019-nCoV genome sequence of the imported cases was 10 (7-24). A total of 134 nucleotide mutation sites were found in all 2019-nCoV genome sequences from 7 entry countries in Gansu province, distributed in 11 open reading frames (ORFs). The top three nucleotide mutations in different proteins: ORF1ab (78), S(20), N(12). Among the 134 nucleotide mutations, 82 caused amino acid mutations, and all of them were missense mutations. No insertions or deletions were seen. Types of deletion mutations, the top three amino acid mutations in different proteins: ORF1ab (46), S(11), N(10); the key sites of the receptor binding domain (RBD) of the S protein have not been mutated.Conclusions:No imported cases in Gansu province have been found to carry the reported mutations that can clearly lead to changes in the spread and pathogenicity of 2019-nCoV.