Cytomegalovirus(CMV) is an important pathogen of congenital and postnatal infections in children,which causes a series of acute and chronic infectious diseases and nervous system sequelaes.Early and accurate diagnosis of pediatric CMV infection is an effective way to improve health in children.This paper will introduce the types,laboratory techniques and diagnostic strategies of CMV infection based on the diagnostic standards at home and abroad,and also focus on current progress in diagnosis of pediatric CMV infection.

Objective To establish a screening system of efficient small interfering RNA (siRNA) target sites directed against truncated region of human cytomegalovirus (HCMV) UL54 gene (UL54S) with siRNA expression vectors. Methods Two small hairpin RNA (shRNA) expression vectors targeting truncated region of HCMV UL54 gene were constructed based on pAVU6 + 27 vector, and cotransfected into AD293 cells with the fusion protein expression vectors pUL54S-enhanced green fluorescent protein (EGFP). The levels of mRNA and EGFP were evaluated by means of reverse transcription-polymerase chain reaction (RT-PCR), fluorescence microscopy and flow cytometry so as to assess the inhibitory efficiency of siRNA. Analysis of variance was applied to analyze the variance of total fluorescence intensity to screen out the efficient target sites of siRNA.Results shRNA expression vector psiUL54-2 and fusion protein expression vector pUL54S-EGFP were cotransfected into AD293 cells. The EGFP expression level in pUL54S-EGFP/psiUL54-2 cotransfected group was lower than that in pUL54S-EGFP/pAVU6 +27 cotransfected group after 48 h of transfection. Gel analysis showed that the mRNA relative level of UL54S was 19.6 after 48 h of psiUL54-2/pUL54S-EGFP cotransfection, which was significantly lower than those in pUL54S-EGFP/psiUL54-1 group (96.6) and control group (100.0). Cotransfection of psiUL54-1/pUL54S-EGFP for 48h didn't show any effects on the expression of fusion protein UL54S-EGFP (P>0. 05).While psiUL54-1/pUL54S-EGFP cotransfection inhibited the expression of fusion protein UL54S-EGFP(19.43×104±2.29×104vs27.89×104±5.50×104, P<0.01).Conclusion Thescreening system of efficient siRNA targeting truncated region of HCMV UL54S is established successfully. The 1532th-1550th nucleotide acids of UL54S coding sequence are efficient siRNA target sites.

Objective To investigate the association between genetic polymorphisms and protein levels of mannose-binding lectin (MBL) and the sensitivities of common infections in a pediatric Han population lived in Zhejiang Province.Methods MBL genetic polymorphisms of patients and controls were detected by PCR-based sequencing.MBL protein levels were measured using MBL ELISA Kit.Results No mutations at positions +223 and +239 of the exon 1 were detected in either patients or controls.No mutation at position +4 of the promoter was detected in controls.The frequencies of the three genotypes HH,HL,and LL at position-550 of the promoter were different between patients and controls(P＜0.05).The frequencies of genotypes YA and XB relevant to MBL protein levels were also different between patients and controls(P＜0.05).Comparing the frequencies of genotypes YA and XB in separate infectious disease with controls,significant differences were found in the group of RRI and CMV infection.The distributions of serum MBL level frequencies in patients and controls were both characterized by skewed distributions.MBL levels of patients with CMV infection were lower than those of controls(P＜0.05).Inversely,MBL levels of patients with acute respiratory infection and localized abscess were higher than those of controls (P＜O.05).Conclusion Genetic polymorphism of MBL gene is seemed to be relative to the sensitivity of common infections in children.

The human cytomegalovirus (HCMV) is a widespread herpesvirus. Virus reactivation from latency can lead to stillbirth, miscarriage, fetal anomalies, and intrauterine growth retardation. During latent infection with the HCMV, the virus can be cleared by the immune response or apoptosis of host cells. However, the HCMV has developed several strategies to manipulate expression of its genes and the microenvironment of host cells. Recent studies have shown that latent infection with the HCMV is associated with viral: regulation of early expression of genes; evasion of cell death; evasion of the immune response; regulatin of non-coding RNAs. This review summarizes recent research progress on the mechanisms underpinning latent infection with the HCMV.
Animals ; Cytomegalovirus ; genetics ; physiology ; Cytomegalovirus Infections ; immunology ; virology ; Humans ; Viral Proteins ; genetics ; metabolism ; Virus Latency

OBJECTIVE Study the role of estrogen receptor (ER)in the inhibition of cell viability and differentiation induced by bisphenol A (BPA)in micro mass culture of rat e mbryonic midbrain(MB) cells.METHODS Micro mass cultures of MB were prepared fro m rat e mbryonic midbrain on gestation day 13.MB cells were exposed to BPA (10 -4 ,10 -6 ,10 -8 ,10 -10 ,10 -12 mol·L -1 )for 5 d.Cell viability was assessed by neutral red uptake test.MB differentiation was detected by he matoxylin staining and i mage analysis.In order to observe the role of ER pathway in the toxicity induced by BPA,cell cultures were co-treated with ICI182780 0.1 n mol·L -1 ,ta moxifen 1 n mol·L -1 and BPA 0.1 mmol·L -1 for 5 d, the cell viability and foci differentiation were detected.Moreover,the protein expression levels of ER in normal e mbryonic brain of gestation day 18,testis tissue fro m adult rats and midbrain cells untreated with BPA were investigated by Western blot.The mRNA expression levels of ER in normal e mbryonic brain of gestation day 13 and gestation day 18,ovary and testis tissue fro m adult rats,and midbrain cells un-treated with BPA were investigated by real-ti me PCR.The mRNA expression levels of Notch1 and Hes1 in MB cells treated with BPA 0.1 mmol·L -1 were also detected by real-ti me PCR.RESULTS BPA 0.1 mmol·L -1 could inhibited MB cell viability and foci differentiation.However,this effect could not be reversed by ER antagonist.The protein and mRNA expression levels of ER in e mbryonic brain and MB cells untreated with BPA were found to be extre mly low.In addition,BPA 0.1 mmol·L -1 could inhibited the mRNA expression levels of Notch1 and Hes1 .CONCLUSION BPA could inhibited MB cell viability and foci differentiation.ER pathway might be not involved in this effect.Instead,Notch-Hes pathway might be involved for this effect.

OBJECTIVETo analyze genotypic diversity of oral Streptococcus mutans (S. mutans) and find out horizontal transmission possibility of the microbe in day-nursery children.

METHODSThe plaque samples were scratched with sterilized toothpicks from teeth of 32 day-nursery children aged between 3 and 4, then cultured on MSB plates. Clones with representative S. mutans-like were subcultured and identified to species level biochemically. AP-PCR fingerprinting was performed to distinguish genotypic diversity of those isolates. Then S. mutans isolated from different children with very similar amplicon profiles were examined by chromosomal DNA fingerprinting analysis.

RESULTSS. mutans were isolated in oral cavities of 78.1% children, 100% in caries and 69.6% in caries-free children. A total of 57 genotypes were identified by AP-PCR. More than one amplitypes were identified in 88% of the 25 children with S. mutans colonization. Two pair of children shared common genotypic S. mutans.

CONCLUSIONThere is no evident relation between number of genotype detected and caries. The presence of matching genotypes of S. mutans among day-nursery children suggests the horizontal transmission may exist.

OBJECTIVETo compare the impacts of different beverages on plaque pH of caries-sensitive and caries-free children and to evaluate the applicability of in situ pH measurements in human dental plaque using Beetrode microelectrode.

METHODSThe subjective population consisted of 20 children (aged 3-5 years). Ten of them were caries-free(dmft = 0); the other were caries-active (dmft > 4). The dental plaque pH were measured in situ with a pH microelectrode within 1 h after drinking three different beverages respectively. Then the resting pH value (pHrest), minimum pH value (pHmin), the range of the pH (deltapH) was analyzed by ANOVA. Results There was a significant difference in pHrest between caries-free and caries-sensitive children. All the pH responses in the plaque following drinking three different beverages showed a classic Stephan-type response. The differences of the pHmin, deltapH in sound sites were no statistic significance (P > 0.05) after drinking beverages in caries-free children. However the changes in the caries-sensitive group were more pronounced than the non-caries group. There were significant differences on the pHmin, deltapH among different beverages (P < 0.05).

CONCLUSIONThe cariogenicity of plaque in caries-active children was stronger than that of the caries-free group. Different beverages have different potential cariogenicity.

Objective To investigate the clinical characteristics and emergency management of severe hand-foot-mouth disease(HFMD)associated with encephalitis and neurogenic pulmonary edema(NPE)caused by en-terovirus 71(EV71)in children.Method Data of critical patients with severe HFMD associated with encephalitis and NPE admitted to pediatric intensive care unit(PICU)Fuyan city Hospitals Anhni Province from May to June 2008 were reviewed.Results Of 30 patients,the mean age was 15.8 months ranged from 4 months to 48 months.The overall morality was 19.4%.Tha average duration of critical symptoms persisted Was 2.1 days ranged from 12 hours to 5 days.There were no rash found in 12 patients(33.3%).The chinical features of nervous system mani-fested the symptoms of brainstem encephalitis in 27 patients(75%),brainstem encephalitis with myelitis in 6 pa-tients(16.7%),and encephalitis in 3 patients(8.3%).The frothy expectoration tinged with pink or bloody,asyrmmetrical pulmonary edema or hemoptysis were the main features of NPE.The main approaches to the treatment were mechanical ventilation,mannitol,methylpredifiselone,intravenous immunoglobulin(IVIG),and vasoactive a-gents.And nine patients(25%)needed fluid volume resuscitation in addition.Conclusions Young children are particularly vulnerable to the Severe EV71 encephalitis with NPE.The majority of involved fatal patients are aged under 3 years.Patients may die of acute onset of NPE and/or hemoptysis with rapid progress towards cardiopul-monary failure.Early diagnosis and evaluation,respiratory support,lowering intracranial pressure and maintaining hemodynamics ale the essential therapeutic approaches.

OBJECTIVETo find out horizontal transmission of oral Streptococcus mutas (S. mutans) in nursery children through analyzing the similarity of S. mutans genotypes.

METHODSThe study group included 24 nursery children between 3 and 4 years of age. Dental plaque samples were collected with sterile toothpick and cultured on MSB plates for 48 h. Individual Streptococcus mutans group colonies representative of the colonial morphologies were subcultured on TPY plates. These strains were identified to species level biochemically. AP-PCR fingerprinting was preformed after identification. S. mutans isolates from different children with very similar fingerprinting profiles were examined by chromosomal DNA fingerprinting analysis.

RESULTSStreptococcus mutans group were isolated in oral cavities of 66.7% children, 58.3% in caries-free and 75.0% in caries children. A total of 4' S. mutans isolates from 24 subjects were analyzed by AP-PCR, and 29 different amplitypes were identifyied, 45.8% carried tw. genotypes. There were 2 genotypes of S. mutans isolated repeatedly among 12 nursery children.

CONCLUSIONThe presence of matching genotypes of S. mutans among nursery children suggests horizontal transmission.