Objective To investigate the change of intracellular cholesterol lipid metabolism and related gene expression after vascular smooth muscle cells infection by human cytomegalovirus(HCMV).Methods By separating the primary umbilical artery vascular smooth muscle cells with a multiplicity of infection of HCMV attack as the experimental group and the control group a-dopted the DMEM/F12 medium containing 3% bovine serum for simulating infection.The change of cholesterol content in the smooth muscle cells after virus attack was detected and the gene expression gene was detected by the gene expression profiling chip method.Results After vascular smooth muscle cells were infected by HCMV,the intracellular gene expression changed significant-ly,in which low-density lipoprotein receptor-related protein 10,11,12 and HMG-CoA synthase,HMG-CoA reductase and B scaven-ger receptor were upregulated.Apolipoprotein A1 and apolipoprotein M were downregulated.Conclusion HCMV infecting vascular smooth muscle cells may alter the lipid metabolism pathway related gene expression,enhance intracellular cholesterol synthesis,re-sult in the cholesterol metabolism imbalance,thus participate in the pathogenesis process of atherosclerosis.

OBJECTIVE: To promote the rational use of antibiotics.METHODS: A retrospectively analysis was conducted on the application of antibiotics in 4 186 discharge case histories in 2005.RESULTS: Of the total 4 186 cases,the application rate of antibiotics was 64.48%,among which,56.17% were prophylactic use of antibiotics and 5.89% used antibiotics without indication.The consumption of antibiotics occupied 45.90% of the total medicines consumed.The nosocomial infection rate was 4.16%,of which,23.56% were fungous infections.The incidence of adverse drug reactions was 2.56%.CONCLUSION: The rate for the prophylactic use of antibiotics in our hospital is on the high side,which may result in high incidences of drug resistant strains and nosocomial infections,therefore,measures should taken to tight the control of the administration of antibiotics.

Objective: To buildSERPING1-knockout porcine fetal fibroblasts(PFFs) based on CRISPR/Cas9 technology and provide cell experimental materials for the construction of hereditary angioedema models. Methods: Firstly, protein structure prediction software was used to analyze the amino acid homology between human and pigSERPING1. Secondly, the eighth exon was selected as the knockout target by screening the effective coding region of the pigSERPING1gene. A single guide RNA targeting pigSERPING1was designed, synthesized and linked to pX330 vector containing Cas9 endonuclease. G418 was used to obtain the positive monoclonal cells after transfection into PFFs. Finally, the circumstance of CRISPR/Cas9 mediated knockout was assessed by the T7 EN1 enzyme digestion assay and the genotypes of monoclonal cells were identified by sequencing analysis Results: Bioinformatic analysis revealed that theSERPING1protein of human and pig had high homology, amino acid sequence identity reached 65.87%. A vector targetingSERPING1was constructed successfully and transfected into cells. Monoclonal cells with knockoutSERPING1gene were obtained through drug screening. Sequencing confirmed the mutant genotype. Conclusion The human and pigSERPING1sequences and their protein structures are highly homologous, and it is suitable for the construction of disease model. CRISPR/Cas9 expression vectors were constructed to achieve theSERPING1gene targeting in PFFs.SERPING1-knockout monoclonal cells were obtained, which could contribute to the construction of theSERPING1knockout miniature pig model.

OBJECTIVE: To explore the clinical features and genetic basis of a patient with glycogen storage disease type VI (GSD-VI). METHODS: Clinical data of the patient was collected. Genomic DNA was extracted from peripheral blood samples of the proband and his parents. Genetic variants were detected by using whole exome sequencing. Candidate variants were verified by Sanger sequencing followed by bioinformatics analysis. RESULTS: The proband presented fasting hypoglycemia, hepatomegaly, growth retardation, transaminitis, metabolic acidosis and hyperlactatemia. Liver biopsy indicated GSD. Novel compound heterozygous PYGL gene variants (c.2089A>G/c.158_160delACT) were detected in the proband. Compound heterozygosity was confirmed by Sanger sequencing of the patient's genomic DNA. Provean and MutationTaster predicted the two variants as deleterious and the variant sites are highly conserved. CONCLUSION The compound heterozygous variants (c.2089A>G/c.158_160delACT) of PYGL gene probably underlay the GSD in the patient. The two novel variants have expanded the spectrum of PYGL gene variants and provided the basis for genetic counseling of the family.
Child ; Family ; Genetic Testing ; Glycogen Storage Disease Type VI/genetics* ; Humans ; Mutation ; Whole Exome Sequencing