Cell culture plays an important role in virology. It provides a platform for the detection and isolation of viruses as well as for the biochemistry and molecular biology based studies of viruses. In the present work, a new system that could permits multiple (different) cell lines to be simultaneously cultured in one dish was developed. In the system, each cell line was cultured in an isolated zone in the same dish or well and the system is therefore called an isolated co-culture system. The usefulness of this novel approach for virus isolation was demonstrated using a model system based on adenovirus.

OBJECTIVETo study the prevalence of internet addiction disorder (IAD) in middle school students of Hunan and to explore its risk factors.

METHODS5760 middle school students and their parents were sampled at random in Hunan province using two-stage sampling (stratified sampling and cluster sampling) method. The ten-item diagnosis tool for IAD, self-rating depression scale, self-rating anxiety scale, interpersonal sensitivity subscale of symptom checklist, family assessment device, parenting locus of control scale, and self-developed questionnaire were employed together to gather related data. 95% confidence interval (CI) was used to describe the prevalence of IAD. Chi-square and logistic regression tests were employed respectively to compare the differences of IAD prevalence among different subpopulations and to explore the possible influential factors.

RESULTSThe overall prevalence of IAD in middle school students of Hunan was 5.52%, with 95% CI as 4.84-6.20. IAD prevalence rates between males and females, being or not being monitored in the class and among different grades, showed significant differences (P < 0.05). No statistical difference was found between single child or having siblings in the family, being at key middle school or ordinary middle school, or within residential areas, (P > 0.05). Gender, tired of going to school, anxiety, interpersonal sensitivity, peer influence in haunting at internet bars, father's education level, the number of recreational settings in the community etc. were the influencing factors of IAD, with ORs as 0.281, 3.469, 2.318, 1.710, 1.877, 1.456, 1.273 and 0.726 respectively (P < 0.10).

CONCLUSIONThe prevalence of IAD in middle school students of Hunan was moderate compared to the reported prevalence rates in other provinces. Gender and peer influence in haunting at internet bars and other 6 factors were suggested to be correlated with IAD events.

Adolescent ; Behavior, Addictive ; epidemiology ; China ; epidemiology ; Female ; Humans ; Internet ; Logistic Models ; Male ; Prevalence ; Sex Factors ; Students ; statistics & numerical data

Adult ; Hemochromatosis ; etiology ; Hepatitis B, Chronic ; complications ; Humans ; Male

Objective To investigate the effect of bundle of care in the prevention of lower extremities deep venous thrombosis ( DVT) among patients with severe ischemic stroke .Methods Two hundred and twenty-eight patients with severe ischemic stroke in the Neurological Intensive Care Unit (NICU) in 2012 were selected as experimental group , and 206 patients with severe acute ischemic stroke in NICU in 2011 as control group.In the experimental group, the patients were treated with bundle of care , which included preventive screening, body position nursing care, nutrition support and liquid management , vascular protection, physical prevention, medicine prevention, passive movement and other measures .And in control group, conventional nursing was applied.The incidence of DVT was observed and compared in two groups .Results There were only 8 patients (3.51%) with lower extremities DVT in the experimental group , and 49 patients (23.79%) with lower extremities DVT in the control group , the difference was statistically significant between the two groups (χ2 =39.004 1,P <0.01).However, there was no statistically significant difference in the incidence of unilateral and bilateral limbs paralysis (χ2 =0.152 9,P=0.696).Conclusions The application of bundle of care can effectively reduce the incidence of deep venous thrombus of lower extremity in patients with severe ischemia stroke.It is worth for clinical use.

OBJECTIVE: To develop a self-administered risk questionnaire for common nutrition-related diseases in middle school students. METHODS: Two phases were conducted to develop the questionnaire: scale development and validation. Phase 1 included 7 steps: (1) determining the objective, theoretical framework, principles and format for indicator generation; (2) setting up the preliminary indicator pool; (3) selecting indicators and forming pilot questionnaire through focus groups; (4) testing the pilot questionnaire; (5) further correcting the questionnaire using expert consultation; (6) choosing indicators again using good-poor analysis; and (7) shaping the final questionnaire. Phase 2 consisted of: (1) using the Pearson correlation coefficient to assess test-retest reliability; (2) using the Cronbach's alpha coefficient to assess the internal consistency reliability; (3) using the feedback from field investigation to assess face validity; and (4) using explanatory factor analysis to assess construct validity. Students from 96 classes were selected at random in Hunan Province as the field test samples using stratified sampling and cluster sampling. And the students from 4 out of the 96 classes were chosen again to serve as the test-retest samples. We used Epidata 3.0 to build the database and SPSS 11.0 to analyze the data. RESULTS: A brief self-administered risk questionnaire for common nutrition-related diseases in middle school students with 12 items being formed after Phase 1. Good-poor analysis showed results from t tests for each item were statistically significant (P<0.05). The Pearson correlation coefficient was 0.76 (P<0.05) and the Cronbach's alpha coefficient was 0.56. The questionnaire was accepted by the students participating in the field test. Four common factors were extracted using explanatory factor analysis, accounting for 50.18% of the total variation. CONCLUSION The brief self-administered risk questionnaire for common nutrition-related diseases in middle school students is reliable and valid.
Adolescent ; Adult ; Child ; China ; epidemiology ; Female ; Humans ; Male ; Malnutrition ; epidemiology ; Nutrition Surveys ; Obesity ; epidemiology ; Risk Assessment ; Self Administration ; methods ; Students ; Surveys and Questionnaires

OBJECTIVETo clone UreB gene of Helicobacter pylori (H. pylori) isolated from children to pGEX-4T-1 expression plasmid, and do sequence analysis.

METHODSA pair of specific primer was designed according to H. pylori UreB gene in the GenBank. Using H. pylori strains isolated from children as a template, a UreB gene was obtained by PCR. After EcoR I and Not I digestion, the PCR production was linked with pGEX-4T-1 which was digested with the same enzymes. The recombinant plasmid was transformed into E.coli BL21 and identified by double enzyme digestion and sequence analysis. The sequence results were compared with the gene sequence in the GenBank.

RESULTSA UreB gene was successfully amplified from children's H. pylori strain GZCH1. It was 1710 bp in size. The objective band was identified by double enzyme digestion. DNA sequence showed that UreB was in the correct open reading frame. The sequence comparison analysis showed that DNA and amino acid sequence identities of UreB gene with other strains were 98%. The sequence of UreB of H. pylori strain GZCH1 was submitted to GenBank (accession number:FJ455126).

CONCLUSIONSUreB of H. pylori strain GZCH1 is successfully cloned to pGEX-4T-1, which provides a basis for research of oral H. pylori vaccine.

Amino Acid Sequence ; Bacterial Vaccines ; immunology ; Child ; Cloning, Molecular ; Helicobacter pylori ; enzymology ; immunology ; Humans ; Male ; Molecular Sequence Data ; Urease ; chemistry ; genetics ; immunology

OBJECTIVETo find out the status of human metapneumovirus (hMPV) infection in children under 14 years old with acute respiratory tract infections (ARI) in Guangzhou, analyze the epidemiology and clinical characteristics among the hMPV-infected children, and provide some basis for research of hMPV.

METHODSAll 521 throat and pharyngeal swabs were collected among the children with acute respiratory tract infections in outpatient departments and those admitted to the wards from September 2006 to August 2008. Then total nucleic acid was extracted from respiratory specimens. The 213 nucleosides of nucleoprotein gene were detected by RT-PCR and 16 strong positive samples were picked to compare with the sequence of hMPV in GenBank after the sequence of the amplification products were determined. Then applied statistical analysis to the data of the collected patients.

RESULTSAll 521 samples were detected by RT-PCR, and confirmed that N gene was positive in 39 samples with a detection rate of 7.49%, and the peak time was in October and April. The 16 amplification products were compared by using the analysis of gene sequence. The nucleocapsid protein (N) gene similarity to BJ1897 of Beijing was up to 99%, and to AY550156 of Thailand was up to 97%, genotype B was the most common genotype.

CONCLUSIONThere existed hMPV infection in children acute respiratory system diseases in Guangzhou areas, in which the children under the age of 6 years were accounted for the main group, however there was no difference in gender. The main symptoms of the patients with hMPV infection were high fever and cough symptom of catarrh. Co-infections other than respiratory virus with hMPV were detected as 41.03% of positive samples.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Male ; Metapneumovirus ; genetics ; isolation & purification ; Molecular Sequence Data ; Nucleocapsid Proteins ; genetics ; RNA, Viral ; genetics ; Respiratory Tract Infections ; epidemiology ; virology

Sodium taurocholate cotransporting polypeptide (NTCP) deficiency is an inborn error of bile acid metabolism caused by mutations of SLC10A1 gene. This paper reports the clinical and genetic features of a patient with this disease. A 3.3-month-old male infant was referred to the hospital with the complaint of jaundiced skin and sclera over 3 months. Physical examination revealed moderate jaundice of the skin and sclera. The liver was palpable 3.5 cm below the right subcostal margin with a medium texture. Serum biochemistry analysis revealed markedly elevated bilirubin (predominantly direct bilirubin) and total bile acids (TBA), as well as decreased 25-OH-VitD level. On pathological analysis of the biopsied liver tissue, hepatocyte ballooning and cholestatic multinucleate giant cells were noted. The lobular architecture was distorted. Infiltration of inflammatory cells, predominantly lymphocytes, was seen in the portal tracts. In response to the anti-inflammatory and liver protective drugs as well as fat-soluble vitamins over 2 months, the bilirubin and transaminases levels were improved markedly while the TBA kept elevated. Because of persisting hypercholanemia on the follow-up, SLC10A1 gene analysis was performed at his age of 17.2 months. The child proved to be a homozygote of the reportedly pathogenic variant c.800C>T (p. Ser267Phe), while the parents were both carriers. NTCP deficiency was thus diagnosed. The infant was followed up until 34.3 months old. He developed well in terms of the anthropometric indices and neurobehavioral milestones. The jaundice disappeared completely. The liver size, texture and function indices all recovered. However, the hypercholanemia persisted, and the long-term outcome needs to be observed.
Humans ; Infant ; Male ; Organic Anion Transporters, Sodium-Dependent ; deficiency ; genetics ; Symporters ; deficiency ; genetics

OBJECTIVETo study dynamics of T lymphocyte subsets in severe acute respiratory syndrome (SARS).

METHODSSequential anti-coagulated blood samples were collected from 46 cases of SARS patients during the 1st week, the 2nd week, the 3rd-5th week and the 8th-12th week after the infection. T lymphocyte subsets including CD3+CD4+ cells, CD3+CD8+ cells, naive CD4+ cells (CD4+CD45RA+CD62L+) and activated CD8+ cells (CD8+CD38+) were detected by 3-color flow cytometry. Fifty-six normal healthy blood donors were also detected as normal controls.

RESULTSCompared with the results of normal controls, both of the percentages of CD4+ cells and CD8+ cells of SARS patients were in normal levels during the 1st week, but the cell counts decreased significantly to (306 +/- 140)/mm3 and (270 +/- 143)/mm3, respectively. The cell count of naive CD4+ subset also remarkably decreased to (96 +/- 49)/mm3, and the percentage of CD8+CD38+ subset was higher than that of normal controls [(59.3 +/- 12.6)% vs (44.9 +/- 12.5)%]. During the 3rd-5th week, the CD8+ cell count and the percentage of CD8+CD38+ subset reached normal values, which were (581 +/- 356)/mm3 and (40.1 +/- 17.6)%, respectively. During the 8th-12th week, the cell counts of CD4+ cell [(578 +/- 193)/mm3] and naive CD4+ subset [(176 +/- 64)/mm3] were still less than those of normal controls, while compared with those of the 1st week, the increments were remarkable.

CONCLUSIONST lymphocytes of SARS patients decreased dramatically but could be obviously resumed in a short time. It will take more than 8-12 weeks for CD4+ cell and naive CD4+ subset to reach to normal levels.

Adult ; CD4 Lymphocyte Count ; CD4-CD8 Ratio ; CD4-Positive T-Lymphocytes ; immunology ; Female ; Humans ; Male ; Middle Aged ; Severe Acute Respiratory Syndrome ; immunology ; T-Lymphocyte Subsets ; immunology

Objective To explore the causative factors of children with respiratory tract infection and their clinical features.Methods Nasopharyngeal aspirate (NPA) was collected from 1746 children with acute respiratory infections (ARIs),aged from 37 days to 3 years,who were admitted to Children's Hospital of Chongqing Medical University between Jun.2009 and May 2011.Fourteen respiratory viruses in NPA were investigated using polymerase chain reactions (PCR) after nucleic acids extracted from the samples and then synthesized cDNA.Sequence analysis was performed to verify the results of each virus from random positive samples.A total of 23 cases were identified with twice or more admissions to hospital.Individual factors,virus detection and clinical characte-ristics were compared between patients with and without repeated hospitalization.Risk factors for repeated hospitalization were investigated.Then these factors were compared in patients with repeated hospitalization between first time and second time.Results The median age was 7 months in patients with repeated hospitalization and 8 months in patients without repeated hospitalization,there were significantly different difference (P ＜ 0.0001).There was no significant difference in virus detection including influenza type A,parainfluenza,respiratory syncytial virus (RSV),adenovirus and human bocavirus between patients with and without repeated hospitalization in the same age group.By comparing clinical features between patients with and without repeated hospitalizatin in the same age group,(69.6％ vs 47.4％,P =O.037) and diarrhea (52.2％vs 29.0％,P =0.017) were found more often in patients with repeated hospitalization.There was no connection between respiratory virus detection and presence of fever,wheeze,cough and diarrhea in patients with repeated hospitalization for the first time of hospitalization.RSV was the most frequently appeared virus in twice hospitalizations,presenting 52.2％ and 30.4％,respectively.One case had twice positive for RSV.In the first hospitalization,there were 16 viruspositive cases and 16 wheeze cases,while in the second hospitalization,there were 12 virus-positive cases (75.0％)and 11 wheeze cases (68.8％),respectively.Conclusions Children with respiratory tract infections suffered repeated hospitalization,mainly occur in children less than 3 years old.Wheeze is an important clinical feature in children with repeated hospitalization.Although causative viruses for patients in each infection are different,RSV is the most common detected viral agent.Virus-positive cases in the first hospitalization present higher viral detection rate during the second hospitalization,with similar trend for wheeze.