ObjectiveTo generate the prokaryotic expression systems of vwA1 and vwA2 genes of Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai,and to determine the correlation among the target recombinant expressed products rVwA1 and rVwA2 and platelet-associated hemorrhage.MethodsThe vwA1 and vwA2 genes of L.interrogans strain Lai were amplified using high fertility PCRs and then the amplification products were sequenced.The prokaryotic expression systems of vwA1 and vwA2 genes were generated by using routine genetic techniques.The expression and dissolubility of rVwA1 and rVwA2 proteins were examined by SDS-PAGE plus Bio-Rad Gel Image Analyzer.Ni-NTA affinity chromatography was used to extract the expressed rVwA1 and rVwA2.Real-time fluorescence quantitative RT-PCRs were performed to determine the changes of vwA1-mRNA and vwA2-mRNA levels in the leptospires of L.interrogans strain Lai before and after infection of human umbilical vein endothelial cell line (HUVEC).The ability of leptospiral rVwA1 binding to human platelets was detected by flow cytometry.A hydrolytic test plus SDS-PAGE were applied to examine the cleavage of leptospiral rVwA2 by a human von Willebrand factor lysase (ADAMTS13).ResultsThe nucleotide and putative amino acid sequences of the cloned leptospiral vwA1 and vwA2 genes were 100％ identities compared to the reported corresponding genes.The generated prokaryotic expression systems of vwA1 and vwA2 genes could express soluble rVwA1 and rVwA2,respectively.When L.interrogans strain Lai infected HUVEC for 8 h,both the vwA1-mRNA and vwA2-mRNA levels were significantly up-regulated (P＜0.05).The result of flow cytometry showed that the leptospiral rVwA1 was able to combine with human platelets with a 60.8％ binding rate. Human recombinant vWF-A2 ( rhvWF - A 2 ),but not the leptospiral rVwA 2,could be hydrolyzed by human ADAMTS 13.Conclusion The vwA1 and vwA2 genes may play roles during infection of L.interrogans species,and the function of vwA1 gene product is referring to the hemorrhage in leptospirosis.

Objective To study on the effect of nttrsing intervention on coping with fatigue in patients with cervical spine fracture and high paraplegia.Methods 46 patients who suffered cervical spine fracture with paraplegia were divided into the control group and the intervention group with 23 patients in each group randomly.The control group received routine care in general,while the intervention group was given additional systematic nursing intervention by full-time nurses.The brief fatigue inventory(FSI),as fatigue assessment tools,was used to assess the fatigue of patients in the two groups before invention and 20 days after invention respectively.Results As a result,we found that the difference of fatigue between the two groups had no prominent significance before intervention,but after invention the difference had prominent significance.Conclusions The nursing intervention is very important for patients with cervical spine fracture and paraplegia and has significant advantages to alleviate or eliminate patients' fatigue and improve their quality of life.

Objective:To investigate the expressions and significance of tyrosine kinase receptor EphA2 and its ligand ephrinA1 in human malignant gliomas and their correlation with tumor angiogenesis. Methods:The expressions of EphA2, ephrinA1, and CD105-stained microvessel density (MVD) were detected via immunohistochemical assay in 62 glioma tissues and 8 normal brain tissues. The correlation between EphA2 and ephrinA1 expression and microvessel counts in the glioma tissues were assessed. Results:Immunohistochemical staining results revealed that variable levels of EphA2 and MVD expression were significantly higher than that of the normal brain samples. Statistical difference was observed in EphA2 and MVD expressions between human gliomas and normal brain samples (P<0.01). The positive rate of EphA2 and MVD expressions was significantly higher in high-grade gliomas (WHO III-IV) than that in low-grade gliomas (WHO I-II) (P<0.01). EphrinA1 was expressed at low levels in most malignant gliomas, and the increased ephrinA1 expression was associated with lower-grade histology. MVD was significantly positively correlated with EphA2 expression (r=0.713, P<0.01) and significantly negatively correlated with ephrinA1 expression (r=-0. 772, P<0.01). EphA2 was significantly negatively correlated with ephrinA1 expression (r=-0.912, P<0.01). Conclusion:Specifically over-expressed EphA2 and its low-expressed ligand ephrinA1 in malignant gliomas may be closely correlated with the invasion and malignant degree of gliomas. Cooperation is involved in the angiogenesis and has an important function in the initiation and progression of gliomas.

OBJECTIVE: To explore the clinical phenotype and genetic variant in a child with Verheij syndrome (VRJS). METHODS: A child who had presented at the Soochow University Affiliated Children's Hospital and Wujiang District Children's Hospital in July 2022 for "elevated scapula since early childhood" was selected as the study subject. Peripheral blood samples of the child and his parents were collected and subjected to whole exome sequencing. Candidate variant was verified by Sanger sequencing and bioinformatic analysis. RESULTS: The child had manifested elevated scapulae, torticollis, neck asymmetry, facial dysmorphism, dispersed café-au-lait spots, limited mobility of upper limbs and shoulder joints, and intellectual disability. Sequencing revealed that he has harbored a de novo heterozygous c.405dupT (p.Ile136Tyrfs*4) variant of the PUF60 gene. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), this variant was classified as pathogenic (PVS1+PS2_moderate+PM2_supporting). Combined his clinical features and result of genetic testing, the child was diagnosed with VRJS due to variant of the PUF60 gene. CONCLUSION The clinical manifestations of VRJS include facial dysmorphism, intellectual disability, elevated scapulae, vertebral fusion, other skeletal malformations, without significant abnormalities of the heart, kidney, and eyes, which need to be distinguished from Klippel-Feil syndrome. Above finding has expended the mutation spectrum of the PUF60 gene and provided a reference for delineation of the genotype-phenotype correlation of the VRJS.
Child ; Child, Preschool ; Humans ; Male ; Cafe-au-Lait Spots ; Computational Biology ; Genetic Testing ; Genomics ; Intellectual Disability/genetics* ; Mutation