Child ; Child, Preschool ; Enteral Nutrition ; adverse effects ; Female ; Humans ; Male ; Pancreatitis ; etiology ; Sodium Chloride

OBJECTIVETo obtain the monoclonal antibody against hexon protein of human adenovirus.

METHODSBALB/c mice were immunized with purified recombinant hexon protein, and the spleen cells of the mice were isolated and fused with myloma cells. Four hybridoma cell strains were screened by indirect ELISA and cultured, and the sensitivity, specificity and virus neutralizing activity were analyzed with ELISA, Western blotting and neutralizing test.

RESULTSThe mouse ascites produced by these hybridoma cells contained specific monoclonal antibodies against hexon protein of human adenovirus as identified by ELISA and Western blot, and the antibody generated by 4C6 strain showed human adenovirus type 3-neutralizing activity.

CONCLUSIONThe monoclonal antibodies against hexon protein with high specificity have been successfully obtained, and these antibodies can be useful in developing assays for early diagnosis of HAdV3 infection and also in study of therapeutic drugs of the infection.

Adenoviruses, Human ; chemistry ; immunology ; Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibodies, Viral ; biosynthesis ; immunology ; Blotting, Western ; Capsid Proteins ; biosynthesis ; genetics ; immunology ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; Humans ; Hybridomas ; secretion ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; biosynthesis ; immunology

OBJECTIVETo study molecular epidemiology of norovirus (NV) infections, stool specimens collected from children with acute diarrhea were tested by TaqMan real-time reverse transcription polymerase chain reaction (RT-PCR) for the viral specific nucleic acid segments.

METHODSFecal samples from a total of 1260 children who had watery diarrhea seen from December 2006 to December 2007 in Guangzhou were analyzed by real-time RT-PCR. The primers and probes used for rapid detection and typing of NV strain target NV sequences were at the ORF1-ORF2 junction, a highly conserved region of the NoV genome. The positive specimens were determined by nested PCR and sequenced.

RESULTSTotally 257 specimens were positive for NV with a positive rate of 20.40%. Shedding of NV type GI was detected in 6.90%, type GII in 16.98% respectively, while the positive number of mixed infection with GI and GII was 44. Of the NV strains that were cloned and sequenced, GI was GI-3, GI-2 and GI-4 detected in positive specimens respectively; meanwhile, GII-4 was most commonly seen in genome II, followed by GII-3 and GII-7. In addition, the average age of children infected with NV was less than 2 years. An epidemic occurred during the winter and early spring (December through the next March).

CONCLUSIONNV was one of the important pathogens for acute diarrhea among children in Guangzhou, which suggested GII-4 was the prevalent strain.

Caliciviridae Infections ; epidemiology ; Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; etiology ; virology ; Feces ; virology ; Humans ; Infant ; Molecular Epidemiology ; Norovirus ; classification ; genetics ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the risk factorsthat predict pain during colonoscopy for decision of sedation or analgesia before the examination.

METHODSA total of 283 consecutive patients undergoing colonoscopicexamination at Nanfang Hospital between July, 2016 and September, 2016were retrospectively analyzed. The clinical data and visual analogue scale after the examination were analyzed to identify the risk factors for pain during colonoscopy using univariate analysis and multivariate logistic regression. A risk stratification model for predicting pain in colonoscopy was established.

RESULTSThe completion rate of the procedure was significantly lower in patients with a visual analogue scale ≥5 (P<0.000). Univariate analysis showed that female patients, previous abdominal surgery, no previous experience with colonoscopy, complaint of abdominal pain before colonoscopy, insufficient experience of the endoscopists, patient's anticipation of high painlevelbefore examination, and a low body mass index (BMI) were all associated with the experience of pain in colonoscopy (P<0.05). Multivariate logistic regressionanalysis identified BMI index (X), level of experience of the endoscopist (A, A, A) and the patient's anticipation of painlevel (X) as the risk factors of pain in colonoscopy(P<0.05), and the establishedmodel with the 3 variables was: P=e/(1+e),Y=0.049-0.124×X-0.97×X+1.713×A+0.781×A+0.147×A, which showed a sensitivity of 70.3% and a specificity of 67.5%for predicting pain in colonoscopy.

CONCLUSIONThe patient's anticipation of a high pain level in colonoscopy, insufficient experience of the endoscopist, and a low BMI are the independent risk factors for pain in colonoscopy, and evaluation of these factors can help in the decision-making concerning the use of sedation or analgesia before colonoscopy.

Abdominal Pain ; etiology ; Analgesia ; Colonoscopy ; adverse effects ; Conscious Sedation ; Female ; Humans ; Male ; Pain Management ; Pain Measurement ; Retrospective Studies ; Risk Factors

OBJECTIVETo understand the etiology of hand, foot and mouth disease (HFMD) in Guangzhou area in 2008.

METHODTotally 1023 clinical specimens were collected from pediatric patients suspected of HFMD in 2008. TaqMan real-time RT-PCR were used for detection of enterovirus 71 (EV71), Coxsackievirus A16 (CA16) and other enteroviruses. The specimens which were enterovirus positive by RT-PCR method with universal primer but EV71 and CA16 negative, were amplified and sequenced for 5'untranslated region.

RESULTEnterovirus was identified from 434 of 1023 samples and detection rate of enterovirus was 42.42%; of the 434 samples, 276 were positive for EV71 (63.6%), 126 for CA16 (29%), 4 samples for enterovirus 84, 3 for Echovirus 11, 2 for Echovirus 9, 3 for Coxsackievirus B3, 4 for Coxsackievirus A10, 3 for Coxsackievirus A6, 6 for Coxsackievirus A12 or A5, and for 7 samples typing was difficult.

CONCLUSIONThe major causative agents of HFMD in Guangzhou were EV71 and CA16 in 2008, and EV84, CA10, CA12, CA6, COSB3, ECHV11, ECHV9 were also the pathogens for smaller proportions of patients.

Child ; Child, Preschool ; China ; epidemiology ; Coxsackievirus Infections ; epidemiology ; DNA Primers ; Enterovirus A, Human ; classification ; genetics ; isolation & purification ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; virology ; Humans ; Infant ; Male ; RNA, Viral ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigative vacA, cagA and iceA genes dominant genotypes of Helicobacter pylori (Hp) isolated from children suffering from gastric and duodenal diseases in Guangzhou area.

METHODSTotally 105 children who underwent gastroscopy in Guangzhou Children's Hospital were enrolled into this study. From each patient, 3 biopsy specimens from the gastric antrum were taken, one was used for rapid urease test, one for histological examination, and one for polymerase chain reaction (PCR) for detecting ureA, vacA, cagA, and iceA genes. DNA was prepared directly from the biopsy specimens from the gastric antrum using a QIAamp DNA mini kit (Qiagen, Germany) according to the manufacturer's instructions. Then 11 primers were used for detecting the genotypes including ureas, (s1, s1a, s1b, s1c, s2) and m (m1, m1T, m2) region of vacA, cagA and iceA (iceA1 and iceA2) genotypes in the 105 children. The distribution of the genotypes of Hp was analyzed.

RESULTAmong the 105 children, only 52 children were positive by the three methods, among these 52 children, 26 were boys and 26 girls. Hp vacA s1as1c/m2 was detected in 43 out of 52 children (82.7%), s1as1c/m1T in 9.6% (5/52), m region that could not betyped was 7.7% (4/52). No strains presented genotypes vacA s1b, s2, m1. The comparison of the positive ratio of vacA s1as1 c/m2 detected in the children infected with Hp and that of the other combination of signal region and middle region was statistically significantly different (P < 0.01). With regard to cagA gene, cagA(+) gene and cagA(-) gene were found in 90.4% (47/52) and 9.6% (5/52) of the children, respectively. The cagA(+) gene was more frequent in the children infected with Hp. Single iceA1 was detected in 78.8% (41/52) children, and single iceA2 was detected to be 1.9% (1/52), multiple strains infection of iceA1 and iceA2 were detected in 3.8% (2/52) children, iceA1 and iceA2 were not detected in 15.4% (8/52), the comparison of the positive ratio of iceA1 detected in the children infected with Hp and that of the other genotypes was statistically significantly different (P < 0.01).

CONCLUSIONThe s1as1c/m2, cagA and iceA1 were the dominant genotypes of Hp in the children in Guangzhou area and s1as1c/m2, cagA and iceA1 were the dominant genotypes combination of Hp in the children in this area.

Antigens, Bacterial ; genetics ; therapeutic use ; Bacterial Outer Membrane Proteins ; genetics ; Bacterial Proteins ; genetics ; Child ; China ; epidemiology ; Female ; Genes, Bacterial ; drug effects ; genetics ; Genotype ; Helicobacter Infections ; drug therapy ; genetics ; Helicobacter pylori ; genetics ; Humans ; Male ; Polymerase Chain Reaction ; Pyloric Antrum ; microbiology

OBJECTIVETo observe the effect of pungent dispersion bitter purgation method (PDBPM) on the esophageal mucosal intercellular space of reflux esophagitis (RE) model rats.

METHODSTotally 100 Wistar rats were randomly divided into the control group, the model group, the Western medicine group (WM), the Chinese medicine group (CM), 25 rats in each group. Rats in the control group only received switch operation. Rats in the rest three groups received modified partial cardia muscle incision combined pylorus ligation of external parts to prepare the RE rat model. Starting from the 3rd day after operation, WM mixture (Motilium 3. 2 mg/kg + Omeprazole Capsule 4.3 mg/kg + Hydrotalcite Tablet 161.4 mg/kg) was administered by gastrogavage to rats in the WM group. Rats in the CM group was administered by gastrogavage with Modified Banxia Xiexin Decoction (5.7 g/kg), 2.5 mL each time, twice daily for 14 consecutive days. Equal volume of normal saline was administered by gastrogavage to rats in the control group and the model group. On day 7 and 14, the lower esophagus pH value, general specimen of mucosa and histopathologic changes were observed. Intercellular spaces of esophageal epithelium were measured for a control study.

RESULTSCompared with the same group at day 7, the lower esophagus pH value increased at day 14 (P < 0.01); the naked eye integral of esophageal mucosa and intercellular spaces of esophageal epithelium also decreased at day 14 in the CM group and the WM group (P < 0.05). Compared with the control group at the same time point, the lower esophagus pH value decreased in the model group (P < 0.01). The naked eye integral of esophageal mucosa, and intercellular spaces of esophageal epithelium increased in the model group with increased intercellular spaces (P < 0.01). Compared with the model group at the same time point, the lower esophagus pH value increased and the naked eye integral of esophageal mucosa decreased in the CM group and the WM group at day 7 and 14 (P < 0.01). Intercellular spaces of esophageal epithelium of RE model rats at day 14 was lower in the CM group and the WM group than in the model group (P < 0.01). Compared with the WM group, the lower esophagus pH value decreased at day 7 in the CM group (P < 0.05); the naked eye integral of esophageal mucosa and intercellular spaces of esophageal epithelium decreased at day 14 in the CM group (P < 0.05).

CONCLUSIONSPDBPM had favorable treatment effect on RE model rats. The therapeutic effect was more obvious along with the therapeutic course went by. Its mechanism might be achieved through good repair effect on damaged mucosa, increasing the pressure of esophageal sphincter, and inhibiting gastric acid.

Animals ; Anti-Ulcer Agents ; pharmacology ; therapeutic use ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Esophagitis, Peptic ; drug therapy ; Extracellular Space ; Mouth Mucosa ; Omeprazole ; therapeutic use ; Rats ; Rats, Wistar

OBJECTIVETo understand the characteristics of human calicivirus (HuCV) infection in infants with diarrhea in Guangzhou city and to study genotype of the virus.

METHODSThe authors collected fecal specimens from 22 children with acute nonbacterial gastroenteritis from November to December, 2001. HuCV was detected from the specimens by RT-PCR. The PCR products were cloned into the PMD18-T cloning vector and sequenced.

RESULTSHCV was detected from the specimens of 2 cases (9%, 2/22). The nucleotide sequence analysis revealed that the virus strains belonged to genotype 2 of Norwalk-like viruses.

CONCLUSIONHuCV is one of the pathogens causing diarrhea in infants and young children in Guangzhou area. HuCV infection occurred sporadically in autumn and winter.

Base Sequence ; Caliciviridae ; genetics ; Caliciviridae Infections ; complications ; virology ; China ; DNA, Viral ; chemistry ; genetics ; Diarrhea, Infantile ; etiology ; Dysentery ; etiology ; Feces ; virology ; Genotype ; Humans ; Infant ; Molecular Sequence Data ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid

OBJECTIVETo survey the clinical epidemiological features of norovirus and rotavirus diarrhea among children living in 5 cities.

METHODA prospective epidemiological investigation was conducted among outpatient children with acute diarrhea between August 2008 and July 2009 in Shanghai, Hangzhou, Guangzhou, Chongqing and Tianjin. The stool samples were randomly collected from children with non-dysentery diarrhea. Group A rotavirus antigen was tested by the kit that applies colloidal gold method in all specimens. GI and GII genogroups of norovirus were detected by one-step real-time reverse-transcription polymerase chain reaction (RT-PCR). The detection rates, seasonality and susceptible age of both viruses-associated diarrhea were analyzed.

RESULTDuring the one-year period, 5091 fecal samples were obtained, of which 1563 (30.7%) were rotavirus-positive. The detection rates of rotavirus were 29.5% (268/916) in Shanghai, 36.1% (334/926) in Hangzhou, 26.3% (254/968) in Guangzhou, 34.1% (359/1054) in Chongqing and 28.2% (348/1233) in Tianjin, respectively. Among the remaining 3528 rotavirus-negative samples, 1049 (29.7%) were identified to be norovirus-positive. The detection rates of norovirus were 21.2%(136/642) in Shanghai, 31.3% (185/592) in Hangzhou, 24.2% (173/714) in Guangzhou, 31.8% (221/695) in Chongqing and 37.7% (334/885) in Tianjin, respectively. It is estimated that the infection rate of norovirus among outpatients with acute diarrhea is at least more than 20.6% (1049/5092). Of 1049 norovirus strains, 1036 (98.7%) were GII genogroup and 16 (1.5%) were GI genogroup, 3 were mixed with GI and GII genogroups. The 1049 children with norovirus diarrhea aged between 1 month and 14 years with the median age of 10 months (mean: 13.9 ± 16.9 months) and 91.8% were 2 years old or younger. The 1563 children with rotavirus diarrhea aged between 1 month and 11.3 years with the median age of 10 months (mean: 12.9 ± 13.7 months) and 92.5% were 2 years old or younger. The median ages between norovirus-infected children and rotavirus-infected children were significantly different (P < 0.05). The peak seasons of rotavirus diarrhea spanned autumn and winter (from October to February). The peak seasons of norovirus diarrhea presented in the winter and spring (from November to April) in Tianjin. Norovirus became active in April and usually predominantly prevalent in the summer and autumn (from July to October) in Shanghai, Hangzhou and Chongqing. However, norovirus was sporadically prevalent in the spring and detected more commonly in the other seasons in Guangzhou.

CONCLUSIONBoth rotavirus and norovirus are the major causative agents for childhood diarrhea. The seasonality of rotavirus diarrhea was similar among the 5 cities. Nevertheless, the seasonality of norovirus diarrhea was diverse in the different areas. In Tianjin located in the north of China, norovirus was quite prevalent in the cold season. In the east (Shanghai and Hangzhou) and south-west (Chongqing), norovirus prevailed dominantly in the summer and autumn. In the south (Guangzhou), the activity of norovirus was more obvious in the summer, autumn and winter.

Adolescent ; Caliciviridae Infections ; epidemiology ; prevention & control ; Child ; Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; prevention & control ; virology ; Female ; Humans ; Infant ; Male ; Norovirus ; Prospective Studies ; Rotavirus ; Rotavirus Infections ; epidemiology ; prevention & control

OBJECTIVETo culture olfactory ensheathing cells (OECs) of rats in vitro and to investigate its morphology, mitosis and immunocytochemistry, and to explore if the OECs could be a new donation for transplantation.

METHODSOECs were harvested from olfactory mucosa of Sprague Dawley rats based on the differing rates of attachment of the various cell types, followed by glial fibrillary acidic protein (GFAP), nerve growth factor (NGF), anti-low affinity receptor for NGF (NGFRp75), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and S-100 immunocytochemistry. The morphological changes and mitosis were observed under a phase contrast microscope at different culture time.

RESULTSThree morphologically distinct types of cells, bipolar, multipolar and flat morphology were present in the primary culture of adult rat olfactory mucosa. Mitosis was characterized by a retraction of all processes, forming a sphere that divided into spherical daughter cells, the daughter cells sent out their processes. The OECs were immunoreactive for GFAP, NGFRp75, S-100, NGF, BDNF and NT-3.

CONCLUSIONSThe OECs from nasal olfactory mucosa cultivated in the medium with fetal bovine serum could survive, divide, differentiate, and express the neurotrophin. It may become an accessible source for autologous grafting in spinal cord injury.

Animals ; Cells, Cultured ; Disease Models, Animal ; Immunohistochemistry ; Male ; Mitosis ; Olfactory Mucosa ; cytology ; transplantation ; Rats ; Rats, Sprague-Dawley