OBJECTIVETo explore the epidemiological characteristics of viral diarrhea of norovirus (NV), sapovirus (SV) and astrovirus (AstV) among children in Zhuhai during winter and spring.

METHODSStool specimens were collected from children with viral diarrhea in Maternal and Child Health Hospital of Zhuhai from November 21, 2009 to April 3, 2010. Nucleic acid of NV, SV and AstV from negative specimens of rotavirus and adenovirus were detected by using Reverse transcription-polymerase chain reaction (RT-PCR), and the types of positive samples of NV were also classified at the same time.

RESULTSThe total detection rate of the three viruses is 21.49 percent, the highest detection rate is 29.05% in December 2009, the lowest detection rate is 12.20% in February 2010, 87.96% of positive specimens were from children patients aged from 0 to 30 months. The season detection rate of NV, SV and AstV are 14.70%, 2.75% and 4.04% respectively. There were significant differences of NV and SV detection rates in every month of the season, whereas the AstV detection rate was comparatively stable. The highest detection rate of NV is 34.09% in children patients aged from 12 to 18 months, the highest SV detection rate is 12.5% in children patients aged from 60 to 120 months, and the highest AstV detection rate is 16.67% in children patients aged from 24 to 30 months. All the NV were belong to G II genogroup.

CONCLUSIONSNV is one of the main pathogens causing viral diarrhea among children in Zhuhai during winter and spring, SV and AstV are also important pathogens. So we should strengthen the monitoring of viral diarrhea caused by NV, SV and AstV in infants and young children.

Child ; Child, Preschool ; Diarrhea ; virology ; Feces ; virology ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Mamastrovirus ; isolation & purification ; Norovirus ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sapovirus ; isolation & purification ; Seasons

In order to develop a rapid detection kit for novel avian influenza virus (AIV) subtype H7N9, two sets of specific primers and probes were designed based on the nucleotide sequences of hemagglutinin antigen (HA) and neuraminidase antigen (NA) of novel H7N9 virus (2013) available in GenBank to establish the method of TaqMan probe-based multiplex real-time RT-PCR for rapid detection of AIV subtype H7N9. The primer and probe of HA were for all H7 subtype AIVs, while the primer and probe of NA were only for novel N9 subtype AIVs. The results showed that this method had high sensitivity and specificity. This method was applicable to the testing of positive standard sample with a minimum concentration of 10 copies/microL; it not only distinguished H7 subtype from H1, H3, H5, H6, and H9 subtypes, but also distinguished novel N9 subtype from traditional N9 subtype. A total of 2700 samples from Zhuhai, China were tested by this method, and the results were as expected. For the advantages of sensitivity and specificity, the method holds promise for wide application.
Animals ; Birds ; virology ; Influenza A Virus, H7N9 Subtype ; genetics ; isolation & purification ; physiology ; Influenza in Birds ; prevention & control ; virology ; Real-Time Polymerase Chain Reaction ; methods ; Species Specificity ; Taq Polymerase ; metabolism ; Time Factors

OBJECTIVETo establish a rapid and convenient method of reverse dot blot (RDB) analysis for detecting the point mutations of non-deletion alpha-thalassemia in Chinese.

METHODSLabel biotin to primers and amplify human alpha2 globin gene selectively, then hybridize PCR products with a set of oligonucleotide probes immobilized on strips, and develop colour to detect non-deletion alpha-thalassemia defects.

RESULTSThe PCR system using biotin-labeled primers could specifically amplify a 1085 bp fragment of human alpha2 globin gene which encompasses all six alpha-thalassemia mutations. After being hybridized with sequence-specific oligonucleotide probes and colour development, it could simultaneously identify all six types of non-deletion alpha-thalassemias encountered in Chinese.

CONCLUSIONThis method does not need semi-nested PCR, and the products amplified by biotinylated primers can be used directly to hybridize with the probes on strips under the identical conditions of hybridization. So, it is a specific and multiplex detection assay for screening non-deletion alpha-thalassemia defects in Chinese.

Humans ; Nucleic Acid Hybridization ; methods ; Point Mutation ; Reproducibility of Results ; alpha-Globins ; genetics ; alpha-Thalassemia ; diagnosis ; genetics

OBJECTIVETo evaluate the expression of epidermal growth factor receptor (EGFR) gene copy number and the expression of ERCC1 and BRCA1 proteins in patients with non-small-cell lung cancer (NSCLC) and the correlation between them.

METHODSThe status of EGFR gene copy number was determined by in situ hybridization (FISH), and the expression of ERCC1 and BRCC1 proteins was examined by immunohistochemistry (IHC). The relationship of EGFR gene copy number with the expression of ERCC1 and BRCA1 and the clinical pathologic features were analyzed.

RESULTSFISH-positive EGFR expression was identified in 40 of 166 samples (24.1%). More FISH-positive EGFR in the female than male patients (31.9% vs. 18.6%, P = 0.048), and non-smoker than smoker (32.8% vs. 16.7%, P = 0.045). FISH-positive EGFR was not associated with age, pathological type, clinical stage and metestasis status (P > 0.05). The expression of ERCC1 protein was identified in 60 of 132 samples (45.5%). The expression of ERCC1 protein varied significantly in tumors of different pathological types (P = 0.046), but not associated with age, gender, clinical stage, metestatic status and smoking status (P > 0.05). The expression of BRCA1 protein was identified in 46 of 131 samples (35.1%). The expression of BRCA1 was not associated with age gender, pathological type, clinical stage, metestatic ststus and smoking status (P > 0.05). There was a moderate correlation between the expressions of ERCC1 and BRCA1 (r = 0.449, P < 0.001), but EGFR gene copy number was not correlated with the expression of ERCC1 or BRCA1 protein.

CONCLUSIONSFISH-positive EGFR expression is associated with gender and smoking status, but not correlated with the expression of ERCC1 and BRCA1 proteins. There is a moderate correlation between the expressions of ERCC1 and BRCA1.

Adult ; Aged ; Aged, 80 and over ; BRCA1 Protein ; metabolism ; Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; pathology ; DNA-Binding Proteins ; metabolism ; Endonucleases ; metabolism ; Female ; Gene Dosage ; Gene Expression Regulation, Neoplastic ; Genes, erbB-1 ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Sex Factors ; Smoking ; Young Adult

OBJECTIVETo develop a 96-microwell plate DNA diagnostic chip for simultaneous detection of 9 major foodborne bacteria.

METHODSType-specific PCR primers labeled with biotin and oligonucleotide probes were designed according to the conservative genes of 9 major foodborne bacteria Staphylococcus aureus, Salmonella spp., Escherichia coli O157:H7 (Stx1 and Stx2), Shigella spp., Listeria monocytogenes, Bacillus cereus, Yersinia enterocolitica, Vibrio cholerae and Vibrio parahaemolyticus. A one-tube multiplex PCR system for simultaneous amplification of these bacteria was established, and the DNA probes were spotted and immobilized in the wells of the plate in 5x5 array format. Stable hybridization system between PCR products and oligonucleotide probes in the microwell was established after condition optimization. Alkaline phosphatase-conjugated streptavidin and NBT/BCIP were used to detect the hybridized PCR products.

RESULTSTwenty standard bacteria strains were used to validate the 96 microwell plate DNA diagnostic chip and highly specific and stable experiment results were obtained. Using this chip assay, the causal pathogen Staphylococcus aureus was identified within 12 h after the sampling from an incident of food poisoning, and the result was consistent with that obtained using conventional bacterial culture and biochemical identification.

CONCLUSIONThe novel 96 microwell plate DNA diagnostic chip allows rapid, accurate, automated and high-throughput bacterial detection and is especially valuable for quick response to such public health emergencies as food poisoning.

Bacteria ; classification ; genetics ; isolation & purification ; DNA, Bacterial ; analysis ; Food Contamination ; analysis ; Food Microbiology ; methods ; Foodborne Diseases ; microbiology ; Humans ; Oligonucleotide Array Sequence Analysis ; methods

OBJECTIVETo discuss the impacts of moxibustion for regulating spleen and stomach function on the survival quality of the patients of end stage renal disease (ESRD) with maintenance hemodialysis (MHD).

METHODSOne hundred and nine cases of uremia with MHD from 3 hemodialysis centers were randomized into an observation group (58 cases) and a control group (51 cases). The regular hemodialysis and conventional medication were used in the two groups. In the observation group, on the basis of the common treatment, moxibustion was applied to Zusanli (ST 36) and Sanyinjiao (SP 6), 2-3 times a day, the treatment of 4 weeks made one session. Totally, 3 sessions were required and the follow-up lasted for 3 months. KDQOL-SF (kidney disease quality of life short form,KDQOL-SFTM 1. 3) was adopted for the questionnaire investigation on survival quality before treatment, after treatment and at the end of follow-up separately in the two groups.

RESULTSAfter treatment, the survival quality scores in terms of physical functioning (83.62+/-13.27 vs 79.32+/- 22. 17), general health (58. 88+/- 20.24 vs 48.82+/-20.89) and vitality (77.07+/-15.56 vs 70. 59+/-22.61) in the observation group were higher than those in the control group (all P<0. 05). In comparison before and after treatment in the same group, the survival quality scores in terms of physical functioning, general health, vitality and symptoms/problems were all improved in the observation group (all P<0. 05). At the end of follow-up, the survival quality scores in terms of physical functioning, general health, mental health, social functioning, vitality, effects of kidney disease and cognitive function were higher in the observation group as compared with those in the control group (all P<0. 05). In comparison of the results at the end of follow-up with those before treatment, the survival quality scores in terms of vitality, symptoms/problems and cognitive function in the observation group were improved (all P< 0. 05). The differences were not significant in all of the 19 fields of survival quality evaluation before and after treatment, and after follow-up in the control group (all P>0. 05).

CONCLUSIONMoxibustion for regulating spleen and stomach function improves the survival quality of the patients with hemodialysis in terms of physical functioning, general health and vitality, which benefits the psychological condition of the patients, resulting in the improvements of the survival quality in the fields of mental health, social functioning, effects of kidney disease and cognitive function.

Adult ; Aged ; Female ; Humans ; Kidney Failure, Chronic ; physiopathology ; therapy ; Male ; Middle Aged ; Moxibustion ; Quality of Life ; Renal Dialysis ; Spleen ; physiopathology ; Stomach ; physiopathology

OBJECTIVETo develop a primer-extension in combination with denaturing high-performance liquid chromatography (PE-DHPLC)-based assay for prenatal diagnosis of the five most common beta-thalassemia mutations in Chinese.

METHODSThe human beta-globin gene fragment was amplified by PCR, followed by a multiple PE reaction specific for each five mutations. Then the PE product mixtures were separated for genotyping of beta-globin gene mutations using fully-denaturing DHPLC analysis.

RESULTSIn a blind study, prenatal diagnosis was performed on thirty-six at-risk families for beta-thalassemia major. Reverse dot blot (RDB) analysis was used to validate each result, showing an accuracy rate of 100% for PE-DHPLC in a total of 108 samples tested. Overall, by PE-DHPLC analysis, the authors could identify the genotypes involving the five mutations and normal alleles corresponding to 94.4% (102/108) and actually make final decision for prenatal diagnosis covering 97.2% (35/36).

CONCLUSIONThe PE-DHPLC protocol can be a simple, rapid, and highly accurate assay in the prenatal detection of common beta-thalassemia mutations.

Base Sequence ; Chromatography, High Pressure Liquid ; methods ; DNA Mutational Analysis ; methods ; DNA Primers ; Female ; Fetal Diseases ; diagnosis ; genetics ; Genotype ; Globins ; genetics ; Humans ; Molecular Sequence Data ; Point Mutation ; Pregnancy ; Prenatal Diagnosis ; beta-Thalassemia ; diagnosis ; genetics

OBJECTIVETo developed a multiplex RT-PCR assay for simultaneous screening of type A, B and novel A (H1N1) influenza viruses.

METHODSTwo pairs of universal primers in were designed for amplifying the M gene and NS gene of type A and B influenza viruses, respectively. A pair of specific primers of HA gene was designed to detect novel A (H1N1) influenza virus. A one-step method was used to establish the multiplex RT-PCR system. A blinded experiment was carried out to validate the accuracy of this assay in comparison with the results of real-time fluorescence RT-PCR. The clinical practicability and efficacy of this assay was also evaluated.

RESULTSThe RT-PCR products were analyzed using agarose gel electrophoresis, which yielded distinct bands of the target fragments without non-specific reactions, suggesting the high efficiency and specificity of the multiplex RT-PCR. Blinded study of 50 samples demonstrated a concordance rate of 100%.

CONCLUSIONThis multiplex RT-PCR assay allows one-step simultaneous detection of type A, B and novel A (H1N1) influenza viruses rapidly and accurately, and provides a valuable low-cost screening technique for influenza epidemic monitoring and early diagnosis.

Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; isolation & purification ; Influenza B virus ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Time Factors ; Viral Matrix Proteins ; genetics ; Viral Nonstructural Proteins ; genetics

OBJECTIVETo explore the pathological basis of diffusion-weighted imaging (DWI) findings in hepatocellular carcinoma (HCC) after transcatheter arterial chemoembolization (TACE).

METHODSDWI was performed in 15 patients with HCC treated by TACE within 24 - 48 hours before II-phase operation. The DWI findings of the liver lesions were analyzed and correlated with pathological findings including macroscopic observation, HE staining and immunohistochemical staining for bFGF.

RESULTS(1) The viable tumor area showed mostly hypersignal intensity (12/15), whereas coagulative necrotic lesions showed hyposignal (8/15) or isosignal intensity (6/15). The ADC values of zones of viable tumor and necrosis in tumor were (1.42 +/- 0.16) x 10(-3) mm(2)/s and (1.58 +/- 0.18) x 10(-3) mm(2)/s, respectively. There was a significant difference of ADC values between the two zones (t = 2.618, P < 0.05). (2) There was a significant difference in ADC values of viable tumor between well and poorly differentiated tumors (t = -2.646, P < 0.05). The distinction of ADC values of the whole tumor was significant among tumors with different degree of necrosis (chi(2) = 7.236, P < 0.05). (3) A negative correlation was observed between bFGF protein expression index and ADC values of viable parts of the tumors in the study group (r = -0.552, P = 0.033).

CONCLUSIONDWI shows certain characteristic features of the HCC after TACE, and can be used to distinguish viable and necrotic tumor tissues in HCC after TACE.

Adolescent ; Adult ; Carcinoma, Hepatocellular ; metabolism ; pathology ; therapy ; Chemoembolization, Therapeutic ; Cisplatin ; administration & dosage ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Fibroblast Growth Factor 2 ; metabolism ; Fluorouracil ; administration & dosage ; Humans ; Iodized Oil ; therapeutic use ; Liver Neoplasms ; metabolism ; pathology ; therapy ; Male ; Middle Aged ; Mitomycin ; administration & dosage ; Young Adult

OBJECTIVETo identify a rare transcription mutation (C-->T) at position -90 of the beta-globin gene previously unreported in the beta-thalassemia carriers from a Chinese family.

METHODSIn phenotype analysis, standard hematological techniques were used to measure RBC counts and Hb concentration. Reverse dot blot (RDB) analysis, which can simultaneously detect 18 known types of beta-thalassemia mutations in Chinese, was used to scan beta-globin gene mutations. DNA sequence analysis of the entire human beta-globin gene was performed to characterize the underlying causative mutation of the sample and to identify its genotype. A semi-quantitative RT-PCR method was used to measure beta-globin gene expression in the form of mRNA from the subjects.

RESULTSThe proband, his brother and his mother presented a typical beta-thalassemic trait with reduced mean corpuscular volume (MCV, 68.2-73.6 fL) and elevated level of Hb A(2) (5.7%-6.4%) but no known beta-thalassemia mutations were found in the samples by RDB analysis. DNA sequencing of the beta-gene region of these three samples revealed heterozygosity for the C-->T substitution at position -90 within proximal CACCC box of the beta-globin gene promoter element, which was previously unreported in the Chinese population. Analysis of mRNA from the positive carriers demonstrated that the mutant beta-globin gene significantly reduced beta-globin transcription (mutants: 2.233 +/- 0.01 vs normal: 3.779+/-1.19; 95%CI: 3.060, 4.499), showing a level comparable with that of the other beta-thalassemia heterozygotes (2.110+/-0.53, 95%CI: 1.732, 2.488).

CONCLUSIONA rare transcriptional mutation that led to beta-thalassemia in Chinese population has been characterized. The findings enrich knowledge of the mutation spectrum of beta-thalassemia.

Adult ; Female ; Globins ; genetics ; Humans ; Mutation ; Transcription, Genetic ; beta-Thalassemia ; genetics