Benefits and related problems of influenza immunization for health care workers / 中华实用儿科临床杂志

Vaccination is the most effective way to prevent influenza and severe outcoming caused by influenza viruses.Health care workers(HCW) are exposed to patients with influenza and they are at high risk of occupationally acquired influenza and of causing nosocomial infection among patients,increasing the incidence rate,the risk of severe and death of patients.Improving the influenza immunization in HCW can not only reduce the prevalence of themselves and keep a weel-oiled of health care facilities during the influenza seasons,but also reduce the risk of severe and death among patients and increase the influenza vaccine uptake in whole population.At present,the influenza immunization coverage of HCW is low.The obstacles and myths of influenza vaccine are barriers for vaccine uptake among HCW.The various strategies are critical in order to improve the influenza coverage rates of HCW.

In vitro expression of human cytomegalovirus UL148 RNA and prediction of its functional sites / 中华微生物学和免疫学杂志

Objective To express UL148 RNA of human cytomegalovirus (HCMV) clinical strains in vitro and to study its functions. Methods Urine of a newborn with HCMV infection was inocula-ted into human embryo lung cells. HCMV clinical strain was isolated and identified by multiplex PCR. UL148 gene was amplified and cloned into pGEM-T-Easy plasmid after double enzyme digestion. A recombi-nant plasmid was constructed and located at the downstream of the T7 promoter. The recombinant plasmid was identified by electrophoresis of the recombinant plasmid,PCR product and double enzyme product. Se-quencing analysis was used for final confirmation. UL148 was transcribed into RNA by 32P labeling. Post-translational modification sites were analyzed by bioinformatics method based on UL148 sequence characteris-tics. Results The clinical strain of HCMV was obtained in vitro. Electrophoresis and sequencing analysis confirmed the successful construction of the recombinant plasmid. UL148 RNA was transcribed in vitro by T7RNA polymerase. Post-translational modification sites showed that UL148 gene contained one cell adhe-sion sequence, one legume lectins beta-chain signature, two N-myristoylation sites, one casein kinase Ⅱphosphorylation site,seven protein kinase C phosphorylation sitse, one cAMP/cGMP-dependent protein ki-nase phosphorylation site, two N-glycosylation sites and one transmembrane region. Conclusion UL148 gene might encode a viral adhesion molecule involving in the signal transduction in host cells.

Effective inhibition of human cytomegalovirus UL148 gene expression by external guide sequences in vitro / 中华实验和临床病毒学杂志

Objective@#To investigate the UL148 gene function of human cytomegalovirus (HCMV) low passage clinic isolate and new strategies for anti-HCMV treatment, the DNA-based external guide sequences (EGSs) were designed to inhibit UL148 RNA expression.@*Methods@#UL148 RNA secondary structure was analyzed by RNA structure technique, an appropriate region was chosen for DNA-based EGS57 synthesis, targeted the UL148 RNA. The M1RNA and UL148 RNA were generated by PCR for transcription in vitro. The UL148 RNA and M1RNA were transcribed in vitro under the function of T7 RNA polymerase. The UL148 was labelled by 32P. The cleavage reactions were carried out by mixing up EGS, M1RNA with UL148 RNA for 1 h. The products were separated by urea denaturing polyacrylamide gel electrophoresis and detected with Typhoon Phosphor Imager.@*Results@#UL148 RNA ranged from 58 to 72 sites was the binding position, and 57 was a cleavage site. EGS57 was designed and synthesized. EGS57 was combined with UL148 RNA to form the natural substrate of M1RNA. UL148 RNA and M1RNA were synthesized through T7 RNA polymerase catalyzing, and the products were conformed. After cleaving reactions, DNA-based EGS57 was shown to be able to cleave UL148 RNA efficiently in vitro by a complex with M1RNA.@*Conclusions@#UL148 RNA was cleaved efficiently by EGS57, and the cleaving site was conformed as expectation. It will provide the gene silent tool effectively for further study the function of UL148 gene.

Analysis the correlation factors of neonatal lenticulostriate vasculopathy / 实用医学杂志

Objective To investigate the correlation factors of neonatal lenticulostriate vasculopathy. Method Four hundred and forty-seven newborns from Guangdong Women and Children Hospital were enrolled in this study. Clinical data of the newborns were obtained . Brain ultrasound studies of lenticulostriate artery were performed on the newborns. The logistic regression was performed for screening the correlation factors of neonatal lenticulostriate vasculopathy (P < 0.05). Results Results of the univariate logistic regression reveal the correlation factors tcontributing to LSV include congenital cytomegalovirus infection、neonatal asphyxia、congenital heart disease (CHD),hypertensive disorder in pregnancy (P < 0.05, respectively). Multivariate logistic regression analysis was performed on these factors. The congenital cytomegalovirus infection, neonatal asphyxia, CHD,hypertensive disorder in pregnancy were significantly associated with LSV (P < 0.05). Conclusion The congenital cytomegalovirus infection,neonatal asphyxia,CHD,hypertensive disorder in pregnancy are the correlation factors of neonatal lenticulostriate vasculopathy. LSV could be a predictive marker for the future development of neuropsychiatric disorders. The brain ultrasound studies of lenticulostriate artery is suggested to be performed on all infants with the correlation factors.

Analysis of nucleotide sequences and genetic polymorphisms of UL138 gene of low passage human cy-tomegalovirus strains in Guangzhou / 中华微生物学和免疫学杂志

Objective To analyze the nucleotide sequences and genetic polymorphisms of UL138 gene of low passage human cytomegalovirus ( HCMV) strains isolated from infants in Guangzhou province. Methods The low passage strains of HCMV were isolated from urine samples of 10 infants with HCMV in-fection in Guangzhou province and identified by multiplex PCR.The UL138 genes were amplified, cloned and identified with sequencing.The sequences were analyzed together with the homologous sequences of 10 clinical isolates published in GenBank.The sequences of UL138 genes were analyzed by using bioinformatics softwares for investigation of the post-translational modification sites, isoelectric points and second structures of UL138 proteins.Results Three low passage strains of HCMV ( D2, D3 and D52) were isolated from in-fants with congenital HCMV infection.The complete sequences of UL138 genes of the three strains were sub-mitted to GenBank after sequencing identification with the GenBank accession numbers of DQ180375, DQ180387 and DQ180359, respectively.The UL138 gene sequences of the three clinical isolates were high-ly conservative.Among the 841 base pairs of the UL138 gene sequences, mutations were identified in 16 sites with base substitution, no any insertion and deletion mutation was found.The 16 mutations resulted in 7 amino acid changes.No additional or deleted sites were found with regard to the post translational modifi-cation sites of UL138 protein in all clinical isolates except the Toledo strain.The isoelectric point of UL138 protein was 6.51 for all clinical isolates.Conclusion The UL138 genes and the deduced amino acid se-quences of HCMV strains isolated from infants in Guangzhou were highly conservative, regardless of the poly-morphism of UL138 gene.This study paved the way for further investigation on HCMV infection and its path-ogenic mechanism.

Human Cytomegalovirus UL143 Gene Polymorphisms in Low-Passage Clinical Isolates in Guangzhou / 热带医学杂志

Objective To investigate the polymorphism of human cytomegalovius UL143 gene of low passage clinical isolates in Guangzhou,China.Method PCR was performed to amplify the entire HCMV ULl43 gene region of 3 clinical isolates,which had been proven by multiplex PCR.The amplification products were cloned into pMD18-T-Vector and subjected to sequencing.The result of DNA sequences were analyzed together with the one of published homologous sequences in GenBank from 14 clinical isolates.Result There were several stop codons in UL143 gene due to a base deletion in open reading frame (ORF) of D3 isolate,which could lead to produce non-functional protein.UL143 ORF of Toledo isolate consisted of 279 nueleotides,encoding a protein with 92 amino acids.UL143 ORFs of other isolates consisted of 252 nueleotides,encoding a protein with 83 amino acids.The DNA sequences were quite conserved and all the variations were base substitution.The amino acid sequences of different isolates were highly conserved.with variation of 1.2%-2.4%.There were no additional or deleted sites of post translational modification of UL143 protein in all clinical isolates except Toledo isolate.There were some differences in the secondary structure among different isolates.The isoelectric point of UL143 protein of all clinical isolates except Toledo isolate was 8.75.Conclusion All DNA and deduced amino acid sequences of UL143 gene shared great similarity among HCMV clinical strains regardless of their polymorphism.