Adult ; Colonic Neoplasms ; genetics ; DNA, Neoplasm ; genetics ; Female ; Fluorescence ; Humans ; Male ; Microsatellite Instability ; Middle Aged ; Polymerase Chain Reaction ; methods ; Rectal Neoplasms ; genetics

OBJECTIVETo investigate the prevalence of human bocavirus (HBoV) in children with acute lower respiratory tract infection and to explore the relationship between the viral load of HBoV and the clinical characteristics of acute lower respiratory tract infection in children.

METHODSA total of 1 554 nasopharyngeal aspirates from children who were hospitalized due to acute lower respiratory tract infection between March 2011 and March 2014 were collected. Quantitative real-time PCR was used to detect 12 RNA and 2 DNA viruses, adenovirus (ADV) and HBoV, and to measure the viral load of HBoV in HBoV-positive children. A comprehensive analysis was performed with reference to clinical symptoms and indicators.

RESULTSIn the 1 554 specimens, 1 212 (77.99%) were positive for viruses, and 275 (17.70%) were HBoV-positive. In HBoV-positive cases, 94.9% were aged <3 years, and there were more males than females. In the 275 HBoV-positive cases, 45 (16.36%) had single infection, and 230 (83.64%) had mixed infection. There was no significant difference in viral load between children with single infection and mixed infection (P>0.05). The patients with fever had a significantly higher viral load than those without fever (P<0.05). The children with wheezing had a significantly higher viral load than those without wheezing (P<0.05). There was no significant difference in viral load between children with mild, moderate, and severe acute lower respiratory tract infection (P>0.05).

CONCLUSIONSHBoV is one of the important pathogens of acute lower respiratory tract infection in children. Children with a higher viral load of HBoV are more likely to experience symptoms such as fever and wheezing. However, the severity of disease and mixed infection are not significantly related to viral load.

Acute Disease ; Adolescent ; Child ; Child, Preschool ; Female ; Human bocavirus ; isolation & purification ; Humans ; Infant ; Infant, Newborn ; Male ; Respiratory Tract Infections ; virology ; Viral Load

OBJECTIVETo establish a rapid, sensitive and specific real-time PCR method for detection of Human Herpesvirus-6 (HHV-6).

METHODSAccording to the reference, a pair of primers and a probe were designed located in U65-66 gene and to set up the standards. We established a real-time RT-PCR method for detection of HHV-6, and to verify the specificity, sensitivity, reproducibility.

RESULTSThe correlation coefficient was 0.999, E = 97.9%, the coefficient of variation values of Ct were 0.61% and 3.13% in real-time PCR assay for inter and intra assay, respectively. The results of all viruses were negative except of HHV-6 for the assay. The quantitative detection limit of the assay was 3 x 10(0) copies/microl.

CONCLUSIONThe real-time PCR assay is highly specific, sensitive and reproducible, which can be used to quatitative detecting clinical samples.

Herpesvirus 6, Human ; genetics ; isolation & purification ; Humans ; Real-Time Polymerase Chain Reaction ; methods ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; methods

OBJECTIVETo discuss the enzyme linked immune spot test (ELISPOT) detected the cellular immune response induced by human Bocavirus (HBoV) VP2 virus-like particles (VLPs).

METHODSAfter immunized by HBoV VP2 VLPs, the specific cellular immune response in mice were detected by ELISPOT assay, observe the ELISPOT results at the conditions of different polypeptide stimulate, different cell culture time, different cell concentration and different specific stimulus peptide concentration, then screening the right ELISPOT experimental conditions and establish the ELISPOT method.

RESULTSThe spots induced by HBoV1 VLPs immunized mice spleen lymphocytes stimulate with polypeptide P3 (GYIPIENEL) and P5 (LYQMPFFLL)were 233 spots/10(6) cells and 157 spots/10(6) cells,spots induced by HBoV2 VLPs immunized mice spleen lymphocytes stimulate with polypeptide P8 (GYIPVIHEL) were 113 spots/10(6) cells; 24 hours is the best time for culture, at this time HBoV1 and HBoV2 groups specificity secretion IFN-gamma ratio were 232 spots/10(6) cells and 119/10(6) cells; Best concentration of mice spleen lymphocyte is 5 x 10(5), right now HBoV1 and HBoV2 group specificity secretion IFN-gamma ratio were 232 spots/10(6) cells and 108/10(6) cells; Best concentration of polypeptides is 10 microg/ml, HBoV1 and HBoV2 group specificity secretion IFN -gamma ratio were 233 spots/10(6) cells and 96/10(6) cells.

CONCLUSIONSHBoV1 and HBoV2 specificT-cell epitope in BABL/c mice were P3, P5 (HBoV1) and P8 (HBoV2). The best experiment condition were: cell cultivated for 24 h, cells concentration for 5 x 10(5) cells/well, stimulating polyperides concentration for 10 microg/ml, it can use to study the cellular immune induced by HBoV in mice.

Amino Acid Sequence ; Animals ; Enzyme-Linked Immunospot Assay ; methods ; Epitopes, T-Lymphocyte ; Human bocavirus ; immunology ; Immunity, Cellular ; Interferon-gamma ; biosynthesis ; Male ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Virion ; immunology

OBJECTIVETo investigate the prevalence of viral pathogen in children with severe pneumonia in Hunan.

METHODBronchoalveolar lavage fluid [BALF] were collected from 122 hospitalized children with severe pneumonia in People's Hospital of Hunan province from January 2011 to December 2011. Nested- or reverse transcription Polymerase chain reaction (PCR or RT-PCR) was used to screen Adenovirus (ADV), Human Bocavirus (HBoV), Parainfluenzaviruses1-4 (PIV1-4), Human Respiratory Syneytial virus (RSV), Influenza virus A (IFVA), Influenza virus B (IFVB), Human Rhinovirus(HRV), Human Metapneumovirus (HMPV), human coronaviruses NL63 and HKU1 (HCoV-NL63, HCoV- HKU1).

RESULTSAmong the 122 bronchoalveolar lavage fluid, viral agents were detected in 60 samples(49.1%), among which ADV (40.98%) was the most common virus, followed by RSV (7.37%) and HBoV (7.37%). Two viruses were detected in 21 individual (35%) samples, of which 20 were dual positive for ADV (40%).

CONCLUSIONADV is the most frequently detected viral etiology of severe pneumonia in children in Hunan during this year. And its Coinfection with other respiratory viruses was common.

Adenoviruses, Human ; isolation & purification ; Adolescent ; Bronchoalveolar Lavage Fluid ; virology ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Pneumonia ; virology ; Reverse Transcriptase Polymerase Chain Reaction ; Seasons ; Viruses ; isolation & purification

UNLABELLEDTo investigate the epidemiological features and types of human adenoviruses (ADV) in children with acute respiratory tract infection in Nanjing area, China. Nasopharyngeal aspirates and nasopharyngeal swabs were collected from 644 outpatients or hospitalized pediatric patients with ARTI at the Children Hospital of Nanjing, Jiangsu Province, China, between August 2010 and July 2011. Adenoviruses were identified and typed from the collected clinical specimens by nested-PCR based on the partial region of the hexon gene. Other 12 respiratory viruses including human bocavirus (HBoV), respiratory syncytial virus (RSV), human rhinovirus (HRV), parainfluenza viruses 1-4 (PIV1-4), influenza virus A/B (IFVA/B), human metapneumovirus (HMPV), human coronavirus NL63 and HKU1 (HCoV-HKU1 and HCoV-NL63) were also identified by PCR method. All PCR positive products were sequenced and phylogenetic analysis was conducted. It was showed that adenoviruses were detected in 171 patients out of 644 (26. 55%) children, 120 (70.18%, 120/171) for ADV3, 16 (9.36%,16/171) for ADV7, 12 (7.02%, 12/171) for ADV1, 10 (5.85%, 10/171) for ADV2, 6 (3.51%, 6/171) for ADV5, 3 (1.75%, 3/171) for ADV6, 3 (1.75%, 3/171) for ADV57, and 1 (0.58%,1/171) for ADV41. ADV infection could occur in any season. There was a higher possibility of ADV infection from April to July in 2011. Most cases (96.49%) were younger than 7 years old. A total of 99 of the 171 ADV-positive children (57.89%) were co-infected with other respiratory viruses. Respiratory syncytial virus (RSV) and human rhinovirus (HRV) were the most common additional respiratory viruses, Lower respiratory tract infections were the most frequent diagnoses made in the hospital, in which there were 52 pneumonia (30.4%) cases.

CONCLUSIONADV is one of the most important pathogens of acute respiratory tract infection in children in Nanjing area, and adenovirus type 3 was the most prevalent serotype. It is important to develop long-term surveillance.

Adenoviridae ; classification ; genetics ; isolation & purification ; Adenoviridae Infections ; epidemiology ; virology ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny ; Respiratory Tract Infections ; epidemiology ; virology

OBJECTIVETo obtain sufficient recombinant VP2 protein of human Bocavirus and establish it's seroepidemiology assying metbord. METHORD: Tbe capsid protein VP2 DNA genes of HBoV1 and 2 were optimized in accordance with tbe usage of the favorite codons in K coil so as to enhance its protein expression in prokaryotic expressing system. The protein was purified by Ni-NTA column, and its antigenicity was determined by Western Blot. Then establish ELISA to detect the specific anti-VP2 IgG antibodies against HBoV1 and 2 in healthy children aged 3-6 years in Nanjing, China.

RESULTSThe recombinant protein 6 x His-VP2 was produced in a larger quantity at 25 degrees C induced by IPTG (1 mmol/L) over night and purified by Ni-NTA column. Seropositive rates of HBoV1 and 2 were 62.2% and 55.5% and their mixed seropositivity was 37%.

CONCLUSIONThe optimizing expression of the capsid protein VP2 from human Bocavirus constructed successfully and get a high yield under certain conditions. The established ELISA could be used to further analyze seroepidemiology of HBoV in China.

Capsid Proteins ; analysis ; genetics ; Child ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Female ; Human bocavirus ; isolation & purification ; Humans ; Male ; Polymerase Chain Reaction ; Seroepidemiologic Studies

OBJECTIVESTo understand the value of measuring neonatal cerebral regional oxygen saturation (rSO2) using near infrared spectroscopy (NIRS) in assessing cerebral oxygenation, to establish the normal range of neonatal cerebral rSO2 and to collect data of the changes of cerebral rSO2 under certain disease status.

METHODSNine large hospitals participated in the multicenter randomized clinical trial from Jan 2007 to Apr 2008. Using the NIRS human tissue oximeter (TSAH-100) independently developed in China, the cerebral rSO2 of 223 normal full-term and 95 otherwise healthy preterm neonates without any special disease, was detected at 1, 2 and 3 days after birth, respectively. The cerebral rSO2 of 102 neonates with diseases which may affect the cerebral oxygenation, was also detected during the severe phases. The pulse oxygen saturation (SpO2) measured at the finger tip, and also the arterial oxygen saturation (SaO2) measured by blood gas analysis, which could indicate the oxygen supply of the whole body, were obtained simultaneously. The correlations among cerebral rSO2, pulse SpO2 and arterial SaO2 were analyzed.

RESULTS(1) The cerebral rSO2 of the normal full-term neonates was (62+/-2)%. Cerebral hypoxia was defined as rSO2 lower than 58%. The cerebral rSO2 of the normal full-terms was steady at 1, 2 and 3 days after birth respectively, without any significant differences among them (F=0.610, P>0.05). The cerebral rSO2 of the neonates with diseases was (55+/-7)%, which was significantly lower than that of the normal full-term neonates (t=15.492, P<0.05). (2) The cerebral rSO2 was positively correlated with the SpO2 (r=0.74, P<0.01) and the SaO2 (r=0.71, P<0.01). (3) Under some special diseases, the changes of cerebral rSO2 was asynchronous with those of the SpO2: (1) For 18 cases under severe cerebral damages or under relatively low hemoglobin concentration, the cerebral rSO2 was significantly low (50%-58%), but the SpO2 was still normal (above 90%). (2) During the recovery of some critically ill neonates, the increase of cerebral rSO2 was lagged as compared with that of pulse SpO2. Especially, during the severe phases of 6 cases with multi-organ failure, the SpO2 and the cerebral rSO2 were both significantly low (55%-80% for SpO2, and 44%-50% for cerebral rSO2); when the diseases were alleviated, although the SpO2 recovered to above 85%, the cerebral rSO2 was still significantly low (around 50%). (3) In 3 cases, during the severe phases of serious hypoxic-ischemic encephalopathy (HIE), the cerebral rSO2 significantly increased to 70%-72%, which was significantly higher than the normal value (62%).

CONCLUSIONSThe range of cerebral rSO2 of the normal full-term neonates was (62+/-2)%. Cerebral oxygenation can be externally indicated by the rSO2 noninvasively and continuously measured by NIRS, which was positively correlated with traditional pulse SpO2 and arterial SaO2. In some special diseases, the rSO2 measured by NIRS can be helpful for clinical diagnoses and treatments.

Birth Weight ; Brain ; metabolism ; Female ; Humans ; Hypoxia, Brain ; diagnosis ; Infant, Newborn ; Male ; Oximetry ; methods ; Oxygen ; analysis ; Spectroscopy, Near-Infrared

OBJECTIVETo study the clinical and laboratory features of the mild and severe hand-foot-mouth diseases (HFMD) in Shenzhen in 2008.

METHODS145 cases were observed in East-Lake Hospital and Shenzhen Children's Hospital. Of the 145 cases, 124 mild cases and 21 severe cases were involved.All the clinical data and laboratory findings were collected and summarized. After collection of the acute and convalescent consecutive stools and peripheral bloods from the patients with HFMDI, EV71 genes were amplified from these samples by RT-PCR. Enterovirus 71 were cultured and isolated using Vero cell line and R&D cell line.

RESULTSThe WBC counts and blood glucose levels of the severe cases were significantly elevated, but the ages of the severe ones significantly decreased compared with those of the mild cases (P < 0.05). EV71 genes could be detected by RT-PCR with 35% positive rate in mild cases and 67% in severe cases. The EV71 gene detection rate of the severe cases was significantly increased in contrast to that of the mild ones. The EV71 were isolated and cultured from the stools of 9 patients, one specimens from the dead's stool. Two severe cases died of neurogenic pulmonary edema and brain-stem encephalitis.

CONCLUSIONSEV71 mainly contributes to HFMD and is responsible for death of some severe cases. High fever, less rash, elevated white blood cell counts and blood glucose concentrations as well as age less than 4 years old should be used for prediction of severe cases.

Adult ; Blood Glucose ; physiology ; Child ; Enterovirus ; isolation & purification ; Enterovirus Infections ; blood ; pathology ; Female ; Hand, Foot and Mouth Disease ; blood ; pathology ; virology ; Humans ; Laboratories ; Leukocyte Count ; statistics & numerical data ; Male ; Reverse Transcriptase Polymerase Chain Reaction ; Severity of Illness Index

OBJECTIVETo estimate the frequency of injections and proportion of unsafe injections and to analyses the critical determinants of poor injection practices in general population in China. Also, to study knowledge, attitudes, practice research in providers and general population.

METHODSA random sample consisting residents and health care providers in a rural county was elected and interview about the frequency of received injection, as well as knowledge, attitudes and practices regarding injections were studied.

RESULTSOverall, 1 004 village residents, and 94 providers were interviewed. Among residents, 145 persons (14.4%), with 457 times (0.46 times per person) had received at least one injection during the previous 3 months. The frequency of injection was 1.84 per year. The proportion of received injections on treatment and immunizations was significantly different among > 12 years age group and < or = 12 years age group. Ninety-four point four percent of disposable syringes/needles were used for injections. Knowledge among the population and providers regarding injection safety was limited.

CONCLUSIONInjections were moderately frequent in this rural area and the proportions of disposable syringes/needles used for injections was very high. Knowledge of safe injection and reasonable injection as well as consciousness of self-protection in the providers and residents need to be improved.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Disposable Equipment ; statistics & numerical data ; Equipment Reuse ; statistics & numerical data ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Infant ; Injections ; adverse effects ; statistics & numerical data ; Male ; Middle Aged ; Risk Factors ; Rural Health Services ; Safety ; Surveys and Questionnaires ; Syringes