OBJECTIVETo develop a method for detection of coxsackie B virus type 1-6 by RT-PCR.

METHODSA pair of primers were designed to amplify all types of coxsackie B virus 1-6 efficiently. The PCR product was hybridized in micro-wells in which 6 type specific oligonucleotide probes had been coated respectively, colorimetric detection was performed to discriminate the types of coxsackie B virus.

RESULTSThis method was shown to be concordant with the IgM ELISA, 71.7% of anti-coxsackie B positive cases could be detected by RT-PCR.

CONCLUSIONThe RT-PCR method can type coxsackie B virus efficiently and provides a tool for clinical diagnosis and epidemiological investigation.

DNA Primers ; Enterovirus B, Human ; classification ; genetics ; isolation & purification ; Enterovirus Infections ; diagnosis ; virology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin M ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; methods

OBJECTIVEBrugada syndrome is an inherited channelopathy that characterized by ST-segment elevation in the right precordial lead (V(1)-V(3)) on the electrocardiogram with or without right bundle branch block and related with high risk of sudden cardiac death and structurally normal hearts. The first and only gene linked to this disease is SCN5A, a gene encodes for alpha subunit of the cardiac sodium channel. The objective of this study is to explore SCN5A gene mutations in Chinese patients with Brugada syndrome.

METHODSFour patients diagnosed as Brugada syndrome and nine patients with suspected Brugada syndrome were chosen for the study. The exons in the functional regions of SCN5A gene were amplified with polymerase chain reaction and the amplified products were sequenced with Sanger method. If a mutation was identified, patient's family members were also screened.

RESULTSTwo heterozygous mutations were found in one family diagnosed as Brugada syndrome. One missense mutation was a G-->A transition in the first nucleotide of codon 95 in SCN5A gene exon 3, which was predicted to result in substitution of Valine with Isoleucine (V95I). The other missense mutation was a C-->T transition in the second nucleotide of codon 1649 in SCN5A gene exon 28, which was predicted to result in substitution of Alanine with Valine (A1649V). A heterozygous mutation was identified in one family suspected to have the disease. The mutation was a three nucleotides (TCT) deletion that caused Phenylalanine deletion in codon 1617 in SCN5A gene exon 28. The three mutations were not detected in 100 control chromosomes.

CONCLUSIONSMutation in SCN5A gene is one of the causes of Brugada syndrome in Chinese. Three novel SCN5A gene mutations were identified in Chinese with Brugada syndrome, which expands the spectrum of SCN5A mutations associated with the disease.

Adolescent ; Adult ; Aged ; Brugada Syndrome ; genetics ; Case-Control Studies ; Exons ; genetics ; Humans ; Male ; Middle Aged ; Muscle Proteins ; genetics ; Mutation ; NAV1.5 Voltage-Gated Sodium Channel ; Sodium Channels ; genetics

There are four different types of N-terminal amino acid sequences (F-I-0, F-I, F-II, F-III) in the multicomponents of earthworm fibrinolytic enzymes (EFE). In GenBank 21 nucleic acid sequences of EFE have been reported. Among them, most of the N-terminal amino acid sequences belong to the F-III type,few belong to the F-II type. Only one is similar to the F-I type, but none to F-I-0. In this research we hoped to obtain the gene encoding component F-I-0 of EFE by the bioinformatics tools. Based on the N-terminal amino acid sequence VVGGSDTTIGQYPHQL of the F-I-0 type from Lumbricus rubellus, a nucleic acid sequence was obtained by in silico cloning from dbEST of Lumbricidae using the software DNAMAN. A new gene of EFE from Eisenia foetida was successfully obtained by RT-PCR using specific primers designed according to this sequence. The new gene named EfP-0 was cloned in pMAL-c2x and expressed as the fusion protein MBP-EfP-0 in the supernatant of lysate. The fusion protein MBP-EfP-0 purified by affinity chromatography had hydrolytic activity on casein plate. Sequencing result shows, EfP-0 has 678bp and encodes a protein of 225 amino acids. The protein is a serine protease belonging to trypsin family. It has similar amino acid composition to F-I-0. BLAST in GenBank shows that the similarity is lower than 40% between EJP-0 gene and other EFE genes. By this we conclude that EfP-0 gene of EFE is a novel gene and it is the first time to be reported, its accession number for Genbank is DQ836917.
Amino Acid Sequence ; Animals ; Cloning, Molecular ; Computational Biology ; Databases, Genetic ; Endopeptidases ; biosynthesis ; chemistry ; genetics ; metabolism ; Escherichia coli ; genetics ; Expressed Sequence Tags ; metabolism ; Molecular Sequence Data ; Oligochaeta ; enzymology ; genetics ; Open Reading Frames ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Objective To investigate the molecular-epidemiologic characteristics and genotypes of human calicivirus (HuCVs) in acute diarrhea patients in Hangzhou from 2009 to 2010.Methods Epidemiologic data and fecal specimens were collected from patients with acute diarrhea.HuCVs of 920 specimens were detected by PCR.PCR products of several positive samples were randomly selected and sequenced.All the sequences were analyzed,phylogenetically.Results 201HuCVs positive cases were identified from 920 facal specimens (21.8％).25 isolates would include norovims G Ⅰ -type,G Ⅱ -type for 170 strains and sapovirus for 1 1 strains.Norovirus G Ⅰ -type and G Ⅱ -type were detected in four specimens at the same time.Other specimens were mixed infection with norovirus G Ⅱ -type and sapovirus.Genotypes of HuCVs showed that norovirus G Ⅰ subtypes were G Ⅰ -1 (3 strains) and G Ⅰ -2 (1 strain).Norovirus G Ⅱ subtypes were G Ⅱ -4/2006b variant strains (7 strains),GⅡ-2 (1 strain),G Ⅱ -7 (1 strain) and G Ⅱ -4/2008 variant strains (2 strains) ;Sapovirus subtypes were G Ⅰ -2 (5 strains),G Ⅰ -1 (4 strains) and G Ⅱ-1 ( 1 strain).The prevalence rates of HuCVs were different in seasons and age groups.Conclusion HuCVs were one of the major pathogens causing acute diarrhea.Both multiple viruses and genotypes of HuCVs were found in the specimens.G Ⅱ-4/2006b variant and similar strains were identified,probably as the prevalent strains from 2009 to 2010 in Hangzhou,Zhejiang province.

OBJECTIVETo prospectively evaluate the change of quality of life in patients with acute coronary syndrome following percutaneous coronary intervention (PCI) with drug-eluting stents and explore the influencing factors of quality of life.

METHODSThere hundred and thirty four consecutive patients with acute coronary syndrome receiving drug-eluting stents implantation between September 2008 and December 2009 were enrolled. Of them, two hundred and ninety three patients completed 36-item short form health survey at baseline and 6 months after PCI procedure. Change of quality of life and influencing factors on quality of life were analyzed.

RESULTSCompared with baseline, quality of life improved significantly after PCI in terms of both physical component summary and mental component summary [ (51.07 ± 20.39) scores vs. (61.69 ± 19.73) scores and (63.27 ± 20.00) scores vs. (68.81 ± 18.71) scores, respectively; all P < 0.01]. Multiple linear regression analysis showed that female, diabetes and ST-segment elevation myocardial infarction were independent predictors of physical component summary improvements post PCI (β values were -0.310, -3.880 and 1.302, respectively; P < 0.05 or P < 0.01). Previous PCI and diabetes were independent predictors of mental component summary improvements post PCI (β values were -1.483 and -2.790, respectively; all P < 0.01).

CONCLUSIONSQuality of life of acute coronary syndrome patients is significantly improved at 6 months after drug-eluting stents implantation. The predictors of physical quality of life improvement are female, diabetes, and ST-segment elevation myocardial infarction. Predictors of mental quality of life improvement are previous PCI and diabetes.

Acute Coronary Syndrome ; surgery ; Aged ; China ; Drug-Eluting Stents ; Female ; Humans ; Male ; Middle Aged ; Quality of Life ; Treatment Outcome

OBJECTIVETo search for the mutations of potassium voltage-gated channel, KQT-like subfamily member 1(KCNQ1) gene in 31 Chinese long QT syndrome(LQTS) families.

METHODSDue to the genetic heterogeneity, the genotype of patients was first predicted based on the spectrum of ST-T-wave patterns on ECG. Ten of 31 probands were considered as LQT1. Then the mutation of KCNQ1 gene was screened by the polymerase chain reaction and single strand conformation polymorphism (PCR-SSCP) technique combined with DNA sequencing in all members of these 10 families. To avoid omitting some LQT1 patients without typical characteristics and also to do methodological comparison, the mutations of KCNQ1 gene on 16 exons were screened by PCR and direct DNA sequencing in the rest 21 non-LQT1 probands only. Co-segregation analysis was carried out after the finding of an abnormal sequence. In case that the abnormality existed in patients only, the test of such exon was performed in 50 irrelevant normal individuals.

RESULTSTwo missense mutations and three single nucleotide polymorphisms (SNPs) were found in the LQT1 predicted families. The two mutations were S277L (1 family) and G306V (1 family) in exon 5 and were not reported previously. Three polymorphisms were 435C-->T (7 families), 1632C-->A (1 family), and IVS1+9 C-->G (3 families). Only a splice mutation IVS1+5G-->A (2 families) and a polymorphism IVS10+18C-->T (1 family) were found in the non-LQT1 predicted probands. All three mutations were localized within the functional domain of KCNQ1 and were co-segregated with the disease, and were not found in 50 normal individuals.

CONCLUSIONTwo novel missense mutations, 1 splice mutation and four SNPs on KCNQ1 gene were found in the 31 LQTS families. Combined with ECG-based genotype prediction, PCR-SSCP could find most mutations on KCNQ1 and be a simple and economic method for screening LQTS.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Humans ; Infant ; KCNQ Potassium Channels ; KCNQ1 Potassium Channel ; Long QT Syndrome ; genetics ; Male ; Middle Aged ; Mutation ; Potassium Channels ; genetics ; Potassium Channels, Voltage-Gated

OBJECTIVEJervell and Lange-Nielsen syndrome (JLNS) is a severe cardioauditory syndrome manifested as QT interval prolongation, abnormal T waves, and relative bradycardia ventricular tachyarrhythmias. In this report, we screened a nonconsanguineous families with JLNS for mutations in KCNQ1.

METHODSMutation analysis was performed by using purified PCR products to direct sequence analysis on an ABI-3730XL automated DNA sequencer. The whole sequence of proband' KCNQ1 was screened firstly, then screened the mutation exon sequences of others of the family and 50 unrelated normal persons.

RESULTSA heterogeneous mutation was identified in the patients of the JLNS family, a missense mutation (G-->T) at nucleotide 917 encoded in exon 6 of KCNQ1. This substitution leads to a change from glycine to Valine at codon 306(G306V) corresponding to the S5 transmembrane segment of KCNQ1. The other normal members of the family and 50 unrelated normal persons were not identified this mutation.

CONCLUSIONThe result suggested that not only homozygous mutations or compound heterozygous mutations in KCNQ1 could cause Jervell-Lange-Nielsen syndrome, the single heterozygous mutation may also cause Jervell-Lange-Nielsen syndrome.

Adolescent ; Adult ; Aged ; Child ; Female ; Genotype ; Humans ; Jervell-Lange Nielsen Syndrome ; genetics ; KCNQ1 Potassium Channel ; genetics ; Long QT Syndrome ; genetics ; Male ; Middle Aged ; Mutation, Missense ; Pedigree ; Young Adult

Objective To explore a new outpatient mode for pregnancy nutrition to adapt to digital hospital.Methods The outpatient management mode and methods were analyzed for pregnancy nutrition.A new outpatient mode combining the technologies of mobile internet and remote monitoring was developed with consideration on standardization,and the effect of the new mode was discussed on pregnancy nutrition outpatient.Results Mobile internet technology and remote monitoring technology contributed to enhancing the efficiency of pregnancy nutrition outpatient,and facilitated the nutrition service of common pregnant women as well as the precision and individualized nutrition management of high-risk ones such as those with gestational diabetes mellitus.Conclusion The new outpatient mode enhances the doctor's efficiency and pregnancy care,and thus is worthy promoting practically.

BACKGROUND: Posterior internal fixation is one of the most common methods for thoracolumbar fractures. There is a lack of systematic evaluation about the efficacy of injured vertebra pedicle screw fixation(IVPSF)versus short-segment pedicle instrumentation (SSPI) for thoracolumbar fracture. OBJECTIVE: To compare the clinical outcomes of IVPSF and SSPI for single thoracolumbar fracture through a METHODS: A computer-based on-line research of PubMed, Medline, Embase, Cochrane Library, CNKI, and WanFang databases was performed for the studies regarding IVPSF versus SSPI for thoracolumbar fracture from 1990 to 2016. meta-analysis. The randomized controlled trials and cohort studies were collected based on the strict criteria of inclusion and exclusion. A meta-analysis was conducted on Revman5.3 sofeware. RESULTS AND CONCLUSION: (1) Eleven articles were enrolled, including 5 English and 6 Chinese ones, involving 689 patients (328 cases for IVPSF and 361 cases for SSPI). (2) The meta-analysis indicated that the operation time, blood loss and mean hospital stay showed no significant differences between two groups. IVPSF showed more effective than SSPI in the kyphotic angle correction and anterior vertebral height recovery at postoperation and 1-5 years of follow-up. Moreover, the incidence of postoperative fixation failure in IVPSF was lower than that in SSPI. (3) These findings suggest that IVPSF that reduces the postoperative fixation failure rate for thoracolumbar fractures provides better kyphosis correction and restoration of anterior vertebral height at post-operation and 1-5 years of follow-up.

Pulmonary arterial hypertension (PAH) is a rare disease with a complex aetiology characterized by elevated pulmonary artery resistance, which leads to progressive right ventricular failure and ultimately death. The aberrant metabolism of arachidonic acid in the pulmonary vasculature plays a central role in the pathogenesis of PAH. The levels of 15-lipoxygenase (15-LO) and 15-hydroxyeicosatetraenoic acid (15-HETE) are elevated in the pulmonary arterial endothelial cells (PAECs), pulmonary smooth muscle cells (PASMCs) and fibroblasts of PAH patients. Under hypoxia condition, 15-LO/15-HETE induces pulmonary artery contraction, promotes the proliferation of PAECs and PASMCs, inhibits apoptosis of PASMCs, promotes fibrosis of pulmonary vessels, and then leads to the occurrence of PAH. Here, we review the research progress on the relationship between 15-LO/15-HETE and hypoxic PAH, in order to clarify the significance of 15-LO/15-HETE in hypoxic PAH.
Arachidonate 15-Lipoxygenase ; Cell Proliferation ; Cells, Cultured ; Endothelial Cells ; Humans ; Hydroxyeicosatetraenoic Acids ; Hypoxia ; Myocytes, Smooth Muscle ; Pulmonary Arterial Hypertension ; Pulmonary Artery