Pediatrics ; organization & administration ; Virology ; organization & administration

Child ; Herpesviridae ; classification ; Herpesviridae Infections ; diagnosis ; Humans ; Paramyxoviridae ; classification ; Paramyxoviridae Infections ; diagnosis ; Parechovirus ; classification ; Pediatrics ; Picornaviridae ; classification ; Picornaviridae Infections ; diagnosis ; SARS Virus ; classification ; Severe Acute Respiratory Syndrome ; diagnosis ; Virus Diseases ; diagnosis ; Viruses ; classification

Adrenal Cortex Hormones ; therapeutic use ; Autoantigens ; immunology ; Child ; Cytokines ; metabolism ; HLA Antigens ; genetics ; Hepatitis, Autoimmune ; diagnosis ; immunology ; therapy ; Humans ; Immunosuppressive Agents ; therapeutic use ; Liver ; drug effects ; metabolism ; pathology ; Liver Transplantation

Objective To investigate the pathogens of acute respiratory infection of children.Methods A total of 159 children with acute respiratory infection who were hospitalized in our department from August 2005 to January 2006 were involved in this study.The serum IgM antibody of 18 pathogens were detected by indirect immunofluorescence test.The 18 pathogens included respiratory syncytial virus(RSV),adenovirus(ADV),influenza A(H1N1,H3N2)and B viruses,parainfluenza viruses(PIV) type 1,2,3 and 4,coxsackie virus B1(CBV1),coxsackie virus A7(CAV7),echovirus(ECHO7),haemophilus influenzae(HI),klebsiella pneumoniae(KP),bordetella pertussis(BP),bordetella parapertussis(BPP) and legionella pneumophila serotype 1 and 12.Results The evidence of specific IgM was obtained in 103 of 159 patients(64.78%).Influenza A was found in 66 cases(64.08%),influenza B in 49 cases(47.57%),enterovirus in 26 cases(25.24%),RSV in 18 cases(17.48%),PIV in 11 cases(10.68%),and co-infection in 66 cases(64.08%),1/ 3 of them were co-infected with influenza A and B.Conclusions Viruses are the most common agents of acute respiratory infection.Influenza virus is predominant among them.

OBJECTIVETo investigate the value of dynamic examination of duodenal fluid in the differential diagnosis of infantile hepatitis syndrome (IHS) and extrahepatic biliary atresia (EHBA). The aim of the study was to establish a simple, rapid and accurate diagnostic procedure for infantile cholestatic jaundice.

METHODSThe authors developed a special duodenal drainage-tube and established a specific duodenal fluid drainage technique. The duodenal fluids were collected and the colors were documented. The bilirubin, gamma-glutamyltranspeptidase (gamma-GT) and bile acid concentrations in the duodenal fluids were measured.

RESULTSDuodenal fluid drainages were initially performed on 561 cases of infants with cholestatic jaundice. The yellow duodenal fluids were drained within 3-8 minutes after intubation in 342 cases. The yellow fluids were obtained in more patients after continuous drainage for 24 hours (21 cases) and 48-72 hours (16 cases), respectively. The duodenal fluids were light yellowish in 71 cases and white in 111 cases. The drainage techniques were subsequently performed in 182 infants with light yellowish or white duodenal fluids after conservative treatment. The duodenal fluids were yellow in 91 cases, white in 89 cases, and slightly yellowish in 2 cases. The increased levels of bilirubin (> or = 8.5 micromol/L), gamma-GT (> 20 IU/L) and bile acid (positive or 33-260 micromol/L) were observed in the yellow duodenal fluids. While the bilirubin levels were 0-2 micromol/L or 5-8 micromol/L in the white or slightly yellowish duodenal fluids, with gamma-GT levels at 0-5 IU/L and bile acid tested negative. According to the criteria set as bilirubin > or = 8.5 micromol/L, bile acid tested positive and gamma-GT > 20 IU/L in duodenal fluid, 470 infants were diagnosed as HIS; 91 cases were diagnosed as EHBA with duodenal fluid bilirubin < 8.5 micromol/L, bile acid tested negative and gamma-GT < 20 IU/L. The diagnoses of these patients were confirmed by surgical operation.

CONCLUSIONDynamic examination of duodenal fluid is a simple, rapid, safe and reliable method in the differential diagnosis of infantile cholestatic jaundice.

Bile Acids and Salts ; analysis ; Bilirubin ; analysis ; Body Fluids ; chemistry ; Diagnosis, Differential ; Duodenum ; metabolism ; Female ; Humans ; Infant ; Infant, Newborn ; Jaundice, Obstructive ; diagnosis ; Male ; Monitoring, Ambulatory ; instrumentation ; methods ; Prognosis ; Reproducibility of Results ; Sensitivity and Specificity ; gamma-Glutamyltransferase ; analysis

OBJECTIVETo investigate the effect of bufalin on nucleus-mitochondria localization of human telomerase reverse transcriptase(hTERT) by exploring its effect on proliferation and apoptosis in human esophageal squamous carcinoma EC9706 cells.

METHODSEC9706 cells were treated with bufalin at various concentrations, and then the cell growth inhibition of EC9706 cells was examined by CCK-8 assay and the 50% inhibitory concentration (IC(50)) was calculated.Cell cycle analysis was performed by flow cytometry with PI staining, and nucleus morphology of apoptosis were observed by fluorescence microscopy with Hoechst 33342 staining. The apoptotic index was measured by flow cytometry with Annexin V-FITC/PI double staining. hTERT subcellular localization and protein expression were determined by Western blotting and multiple immunofluorescence labling combined with laser confocal scanning microscopy.

RESULTSThe proliferation of EC 9706 cells was significantly inhibited by bufalin along with the increase of processing time and concentrations (p<0.01). After the EC9706 cells were exposed to 100 nmol/L bufalin,the number of cells gradually decreased in G(1) phase and increased in S and G(2)/M phases(p<0.05). The typical nucleus morphological changes of apoptosis were observed and the apoptotic index was increased(p<0.01). The expression of hTERT decreased in nucleus but increased in mitochondria(p<0.05).

CONCLUSIONSBufalin can inhibit the proliferation of human esophageal squamous carcinoma EC9706 cells in a time- and dose-dependent manner. It can arrest cell cycle in S and G(2)/M phases and induce the apoptosis of EC 9706 cells. hTERT is localized in both nucleus and mitochondria,and can be partially translocated from nucleus to mitochondria during the bufalin-induced apoptosis.

Apoptosis ; drug effects ; Bufanolides ; pharmacology ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Esophageal Neoplasms ; metabolism ; pathology ; Humans ; Telomerase ; metabolism

OBJECTIVETo investigate the prophylactic, blocking and therapeutic effects of Allitridin on inhibiting HCMV proliferation by measuring the expression level of HCMV IEA in vitro and explore the mechanism of Allitridin anti-HCMVactivity.

METHODSThe cytotocity of Allitridin was evaluated through MTT colorimetry and cell morphology. HCMV IEA levels were quantitatively detected by Flow Cytometry respectively under the following conditions: Allitridin was given before (pretreated for 24 h), during, or after viral inoculation in which serial doses (maximum tolerant concentration, MTC for human embryo lung cells, HEL) of Allitridin was used to treat HCMV infected HLE cells for different durations (24, 48, 72, 96 h) after viral infection.

RESULTThe MTC of Allitridin was 9.60 mg x L(-1). Allitridin remarkably inhibited the expression of HCMV IEA in vitro. Within MTC, the inhibitory rate had a significant correlation with its dosage (r = 0.96). At the time of IEA highest expression (72 h after infection), inhibitory effect was the greatest (inhibitory rate: 89.3%). With pretreatment of Allitridin, the inhibitory rate was 28.6%. When Allitridin was used together with HCMV inoculation, IEA inhibitory rate was only 10.3%.

CONCLUSIONAllitridin can inhibit HCMV, IEA expression in vitro remarkably which is probably one of the major mechanisms of Allitridin anti-HCMV activity because IEAs are the very important regulatory factors for the expression of all HCMV genes. Its therapeutic effect is the best at the peak stage of IE1 gene expression (72 h after infection) but it has low prophylactic and little blocking effect.

Allyl Compounds ; isolation & purification ; pharmacology ; Antiviral Agents ; pharmacology ; Cytomegalovirus ; genetics ; Fibroblasts ; cytology ; metabolism ; virology ; Flow Cytometry ; Garlic ; chemistry ; Gene Expression Regulation, Viral ; drug effects ; Humans ; Immediate-Early Proteins ; metabolism ; Plants, Medicinal ; chemistry ; Sulfides ; isolation & purification ; pharmacology

OBJECTIVETo evaluate the diagnostic potential of previously published enterovirus (EV) reverse transcription polymerase chain reaction (RT-PCR) assay in detection of EV in CSF samples from children with a diagnosis of aseptic meningitis and to investigate the clinical characteristics of the patients seen in Shandong.

METHODSEV RNA was detected in 187 CSF samples and serum and/or urine samples of a part of patients by RT-PCR and viral culture technique.

RESULTSRT-PCR was positive in all 62 CSF specimens which were positive by cell culture (100%). In addition, 93 of 125 (74.4%) CSF samples negative by cell culture were RT-PCR positive. In 4 of these 93 (4.3%) patients, viral culture of specimens from other sites (serum or urine) was also positive. The sensitivity of CSF RT-PCR based on clinical diagnosis in patients with meningitis of negative bacterial culture results was 82.9% (155/187), which was considerably higher than the sensitivity of CSF virus culture 33.2% (62/187). The results of RT-PCR can be reported within 4 hours, whereas the viral culture of CSF requires 4.6 days for a cytopathic effect to develop. EV meningitis occurred in a sporadic form and in some areas there were outbreaks. The clinical characteristics of 155 patients with EV meningitis were different in different age groups.

CONCLUSIONEV was one of the most common causes of aseptic meningitis in Shandong area. The RT-PCR assay was rapid, sensitive and specific for the diagnosis of EV meningitis and may be a potential tests to shorten hospital stay and reduce the use of antibiotics.

Central Nervous System Infections ; blood ; diagnosis ; urine ; Child ; Child, Preschool ; China ; Enterovirus ; genetics ; isolation & purification ; Enterovirus Infections ; cerebrospinal fluid ; diagnosis ; Female ; HeLa Cells ; Humans ; Infant ; Infant, Newborn ; Male ; RNA, Viral ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVEHuman rhinovirus (HRV) is the most common respiratory pathogen, which causes not only acute respiratory infection and community acquired pneumonitis in children, but also asthma episode and deterioration. However, the detection of respiratory pathogen, which mainly focuses on respiratory syncytial virus, influenzaviruses A and B, parainfluenza viruses 1-3 and adenoviruses, does not include HRV yet by now in China. The absence of detection method limits the clinical understanding of HRV pathogenicity, and causes unreasonable use of antibiotics. This study aimed to establish a one-step reverse transcription (RT) PCR system for detecting specific fragment of HRV RNA, and to analyze the sequences of amplicons.

METHODSA pair of degenerate primers based on the HRV highly conserved 5'' noncoding region (NCR) were used to develop a one-step RT-PCR system for detecting HRV RNA in nasopharyngeal aspirates from 78 children with acute respiratory tract infections in the spring of 2004. All the positive PCR products were sequenced, and the sequences of the nucleotides were analyzed by using biological software and compared with those in the GeneBank.

RESULTSEleven (14.1%) of 78 samples were positive on RT-PCR, these patients were clinically diagnozed as upper respiratory tract infection (n = 7), bronchitis (n = 3) and bronchopneumonia (n = 1), respectively. Compared with the sequences of clinical and standard HRV viruses in the GeneBank, the nucleotide sequences of these 11 amplicons shared high homology of 89%-95.5%. Within the 11 amplicons, nucleotide identity varied from 75.2% to 91.8%, and the ratio of genetic variation was from 8.8% to 31.0%, which occurred in highly conserved regions and usually showed single nucleotide mutation in some special locations. These 11 amplicons attribute to the two branches of HRV cladogram, respectively. Most of mutations in highly conservative domain occurred on single ribonucleotide, mainly as transversion (C/G, A/G) and transition (T/C, A/G), some were mutations among 3 bases (A/C/G, A/T/G, A/C/T). And a few mutations involved two nearby ribonucleotide which were also found in highly conservative domain. However, ribonucleotide deletion and insertion were usually found in highly variable domain.

CONCLUSIONThe findings showed that this one-step RT-PCR system was highly specific, rapid and convenient for the detection of HRV RNA in nasopharyngeal secretions of patients with acute respiratory tract infections and that the genome of HRV viruses was highly variable.

Base Sequence ; Child ; Child, Preschool ; Female ; Genes, Viral ; Humans ; Male ; Molecular Sequence Data ; Picornaviridae Infections ; diagnosis ; virology ; RNA, Viral ; analysis ; Respiratory Tract Infections ; virology ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Rhinovirus ; genetics ; isolation & purification ; Sensitivity and Specificity ; Sequence Analysis, DNA

BACKGROUNDThe production of neural stem cells (NSCs) derived from embryonic stem (ES) cells was usually very low according to previous studies, which was a major obstacle for meeting the needs of clinical application. This study aimed at investigating whether astrocytes could promote production of NSCs derived from ES cells in vitro.

METHODSMouse ES cells line-D3 was used to differentiate into NSCs with astrocytes as inducing stromal cells by means of three-stage differentiation procedure. Another group without astrocytes served as control. The totipotency of ES cells was identified by observation of cells' morphology and formation of teratoma in severe combined immunodeficiency disease (SCID) mice. The quantity and purity of NSCs derived from ES cells were analyzed using clonogenic assay, immunohistochemical staining and flow cytometry assay. The plasticity of NSCs was detected by differentiating test. Octamer-binding transcription factor 4 (Oct-4) and nestin, the specific marker genes of ES cells and NSCs respectively, were detected continuously using reverse transcription-polymerase chain reaction (RT-PCR) method to monitor the process of cell differentiation.

RESULTSThe ES cells of D3 line could maintain the ability of differentiating into cellular derivations of all three primary germ layers after continuous passage culture. At the end of two-stage of inducing process, 23.2 +/- 3.5 neurospheres per plate formed in astrocyte-induced group and only 0.8 +/- 0.3 per plate in the control group (clonogenic assay, P < 0.01), and the ratio of nestin positive cells was (50.2 +/- 2.8)% in astrocyte-induced group and only (1.4 +/- 0.5)% in the control group (flow cytometry, P < 0.01). With the induction undergoing, the expression of Oct-4 gradually decreased and then disappeared, while the expression of nestin was increased step by step, and the ratio of nestin positive cells was up to 91.4% by the three-stage differentiation. The nestin positive cells could be further induced into neurons, astrocytes, and oligodendrocytes in differentiating medium supplemented with fetal calf serum. The results of differentiating test showed that the ratio of NF-200 and NSE positive cells was (42.7 +/- 2.6)% in astrocyte-induced group and only (11.2 +/- 1.8)% in the control group (P < 0.01).

CONCLUSIONSAstrocytes can not only increase the production of NSCs derived from ES cells but also promote the differentiation of NSCs toward neuronal lineage.

Animals ; Astrocytes ; physiology ; Cell Differentiation ; Cell Lineage ; Cells, Cultured ; Embryo, Mammalian ; cytology ; Mice ; Neurons ; cytology ; Stem Cells ; cytology