This study aims to analyse the value of CODEHOP RT-PCR in the detection of Flavivirus. According to the amino acid sequences of polyproteins of different flaviviruses published in GenBank, a pair of primers was designed using the CODEHOP method. One-step RT-PCR was used to detect Japanese encephalitis virus strain JEV1201, Dengue virus strain JKD001, and yellow fever virus vaccine YV6161. BLAST analysis and phylogenetic analysis were performed after the RT-PCR products of nucleocapsid genes were sequenced. The results showed that this method could amplify Flavivirus specifically, and the size and sequence of the target fragment accorded with the anticipated result. JEV1201 had the highest homology to Japanese encephalitis virus strain YL2009-4/YC2009-3, belonging to the branch of the phylogenetic tree of Japanese encephalitis virus strains. JKD001 had the highest homology to Dengue virus strain DENV-2/ID/1022DN/1975, belonging to the branch of the phylogenetic tree of Dengue virus strains. YV6161 had the highest homology to Yellow fever virus strain 17D, belonging to the branch of the phylogenetic tree of Yellow fever virus strains. In conclusion, the method of CODEHOP RT-PCR can be effectively used to detect, identify, and phylogenetically analyse Flavivirus.
DNA Primers ; genetics ; Flavivirus ; classification ; genetics ; isolation & purification ; Flavivirus Infections ; virology ; Humans ; Molecular Sequence Data ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Viral Nonstructural Proteins ; genetics

OBJECTIVE: To investigate the expression of hyperpolarization-activated cyclic nucleotide-gated cation channel 4 (HCN4) and connexin43 (Cx43) in the sinoatrial node of electric shock death. METHODS: As experimental group, 34 cases of electric shock death who had definite current mark evidence were selected from pathology department of Xuzhou Medical College from 2010 to 2013. As the control group, 20 cases of fatal severe craniocerebral injury in traffic accidents were chosen. The expressions of HCN4 and Cx43 in the sinoatrial node were observed by immunohistochemical technology. RESULTS: HCN4 positive cells expressed in the cell membrane and cytoplasm of the sinoatrial node. Cx43 positive cells expressed in the cell membrane and cytoplasm of T cells and myocardial cells. The expression of HCN4 was significantly higher than that of the control group (P < 0.05) and the expression of Cx43 was significantly lower than that of the control group (P < 0.05). CONCLUSION The changes of HCN4 and Cx43 expressions in the sinoatrial node illustrate electric shock death might be related to the abnormalities of cardiac electrophysiology and conduction.
Connexin 43/metabolism* ; Cyclic Nucleotide-Gated Cation Channels ; Heart Rate ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels/metabolism* ; Immunohistochemistry/methods* ; Myocardium/metabolism* ; Myocytes, Cardiac ; Sinoatrial Node/physiopathology*

Objective To observe toxic effect of deoxynivalenol(DON)on articular cartilage and synovium of New Zealand rabbits's knee ioints.Methods Fifteen male rabbits were divided randomly into 3 groups:control, high-dosage,and low-dosage group.In high-dosage and low-dosage group,saline solution of DON was injected with a dose of 0.10 and 0.05 ms/kg every 48 h into ear vein of rabbits.Specimen of articular cartilage and synovium were through pathologY methods,and IL-1β,TNF-α,NO levels were assayed in joint liquid,after 20 days. Results Morphological changes were observed, such as synovium inflammative infiltration, chondrocytes deformation and necrosis under light microscope.The levels of IL-1β,TNF-α and NO had statistical significance in comDarison between 3 grouPs(F=19.396,18.195,22.136,P＜0.05).The levels of IL-1β,TNF-α and NO were significantly higher(all P＜0.05),high-dosage[(0.451±0.091),(0.575±0.122)μg/L;(70.27±11.53)μmol/L] and low-dosage group[(0.295±0.107),(0.387±0.131)μg/L;(45.32±12.24)μmol/L]compared with control ((0.1 13±0.049),(0.138±0.087)μg/L;(23.56±9.35)μmoL/L],and high-dosage compared with low-dosage group Conclusions DON results in articular and synovial impairment,which has the symptom similar to osteoarthritis. DON probably causes osteoarthritis.

Aim To investigate the effect of p-hydroxybenzaldehyde ( HD) on intestinal fibrosis in mice based on mouse intestinal fibrosis model and in vitro EMT model,and to explore the underlying mechanism Methods HE staining, Masson staining, immunohisto-chemistry ,qPCR, Western blot and other experimental methods were used to verify the effect of HD on intestinal fibrosis in mice and the potential mechanism. Results In vivo experiments showed that compared with the normal group, the DSS-induced intestinal fibrosis model group had shortened colon, increased colon his-topathological score, increased collagen volume fraction, and significantly increased collagen I expression. After treatment with 4, 10, and 25 mg • kg

OBJECTIVETo evaluate the efficacy and safety of recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) hydrogel in wound healing in patients with deep partial thickness burn.

METHODSThe study was a multicenter, randomized, double-blind, placebo-controlled parallel clinical trial. Three hundred and twenty-one patients (302 cases finally fulfilled the protocol) with deep partial thickness burn were divided into A group (n = 200, with treatment of rhGM-CSF hydrogel, 100 microg/10 g/100 cm2/d), C group (n = 102,with treatment of placebo). Side-effect, systemic condition, wound healing time, wound healing rate, and total effective rate at different time points were observed.

RESULTSThere were no obvious differences in vital signs, wound secretion, wound edge reaction, blood and urine routine, liver and kidney function between two groups (P > 0.05). No side-effect was observed. The median wound healing time was 17 days in A group, which was obviously shorter than that in C group (20 days, P < 0.01). The mean wound healing rate in A group was 24.5%, 70.5%, 95.3%, 99.6% respectively on 8th, 14th, 20th, 28th day after treatment, which were obviously higher than that in C group (15.1%, 51.4%, 84.6%, 97.1%, respectively, P < 0.01). The total effective rates in A group on 8th, 14th, 20th day after treatment were also higher than that in C group (P < 0.01).

CONCLUSIONrhGM-CSF hydrogel can significantly accelerate wound healing in patients with deep partial thickness burn with certain safety.

Burns ; drug therapy ; Double-Blind Method ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; adverse effects ; therapeutic use ; Humans ; Hydrogels ; therapeutic use ; Male ; Placebos ; Recombinant Proteins ; Wound Healing

Objective To determine the prevalence and genotype of hepatitis E virus (HEV)among commercial swine population in Eastern and Southern China. Methods Six hundred specimens of swine bile collected from 5 slaughterhouses in Eastern and Southern China from 2007 to 2009 were tested for HEV RNA using nested RT-PCR. PCR products were sequenced for phylogenetic analysis. Results Forty-seven out of the 600 samples (7.83%) were positive for HEV RNA. Based on the 150 nt fragment within HEV ORF2, data from phylogenetic analysis revealed that all the 47 HEV isolates were identified to be genotype Ⅳ, sharing 75.0%-83.4%, 75.0%-84.6%, 71.9%-80.7% and 88.1%-91.5% nucleotide identities with prototype Ⅰ,Ⅱ,Ⅲ and Ⅳ HEV strains respectively while majority of the isolates clustered within their respective isolation sites. Conclusion HEV was widespread in commercial swine population in Eastern and Southern China that raised a serious concern about the safety regarding the consumption of pork products.

[Objective]To investigate the relationship between clopidogrel response , early neurological deterioration and CYP2C19 gene polymorphism in sICAS patients.[Methods]116 sICAS patients were divided into deterioration group and non-deterio?ration group by whether appear early nervous function deterioration. Record included the baseline data,the genotypes of CYP2C19, platelet maximum aggregation rate after 7 days of given clopidogrel,CYP2C19 genotype and clopidogrel reaction were compared be?tween two groups.[Results]The deterioration group combination with diabetes ,stroke/TIA were significantly higher than the non-de?terioration group(P<0.05);The frequency of CYP2C19*2 AA genotype and A allele were significantly higher than those in non-dete?rioration group. The frequency of GG genotype were significantly lower than non-deterioration group (27.27% vs 2.13%,50.00% vs 14.84%,27.27% vs 72.34%)(P<0.01);The poor metabolic genotype platelet maximum aggregation rate were significantly higher than that of the fast-metabolic genotype and middle metabolic genotype(P<0.01,P<0.05),Middle metabolic genotype platelet maxi?mum aggregation rate were significantly higher than fast-metabolic genotype(P<0.01);The deterioration group fast-metabolic geno?type were significantly lower than non-deterioration group ,poor metabolic genotype were significantly higher than non-deterioration group(22.73% vs 65.96%,36.36% vs 5.32%)(P<0.01);Clopidogrel resistance rate 59.09% were significantly higher non-deteriora?tion group 28.72%(P<0.05).[Conclusion]Clopidogrel response and early neurological deterioration in sICAS patients is associatedwith CYP2C19 gene polymorphism.

Objective To explore the expressions of matrix metalloproteinase-9 (MMP-9) and tissue inhibitors of matrix metalloproteinase-1 (TIMP-1) after focal cerebral ischemia-repeffusion injury in rats. Methods Totally 40 Wistar rats were randomly divided into 2 groups: sham-operated group and iscbemia-reperfusion group. A middle cerebral artery occlusion model was constructed in the 25 rats with Longa's method. The expressions of MMP-9 and TIMP-1 were investigated with immunohistochemistry and HE staining at 6, 24, 48, 72 h and 7 d of reperfusion following 2 h ischemia. Results MMP-9 began to be expressed at 6 h reperfusion, and was obviously increased at 24 h and reached the peak level at 48 h, and then, the expression of MMP-9 began to decrease at 72 h to a low level at 7 d. TIMP-1 positive cells began to arise at 6 h reperfusion, peaked at 24 h, decreased at 48 h and remained a low level at 7 d. The expressions of MMP-9 and TIMP-1 were mainly located in vascular endothelial cells, neurons and gitter cells. The expressions of MMP-9 and TIMP-1 were negative in sham-operated group. Conclusion The expressions of MMP-9 and TIMP-1 are induced to increase by focal cerebral ischemia-reperfusion, and vascular endothelial injury may be the main cause to the high expressions of MMP-9 and TLMP-1.

OBJECTIVETo analyze the spatial distribution of highly pathogenic avian influenza (HPAI) and to explore environmental factors associated with HPAI using geographic information system (GIS) techniques in Mainland China.

METHODSDatabases were set up using the information of HPAI during epidemics in 2004, and linked to digital maps at provincial and county administrative layers in the country through the ArcGIS 8.3 software. Spatial cluster analyses, spatial statistics analyses and tracking analyses on epidemic situation of HPAI were implemented. Environmental factors associated with HPAI were also analyzed on data related to weather, vegetation and migratory birds etc.

RESULTSFindings from spatial cluster analyses showed that high incidence area was centralized in 113.261 degrees ordm; east longitude and 23. 119 degrees ordm; north latitude with a radius of 1090.52 kilometers (relative risk= 2.646, P value= 0.001). Spatial statistical analyses showed that HPAI took place mainly in capital cities of provinces and surrounding areas as well as in the circumference areas of arterial rivers, lakes and seacoasts. Results also showed that HPAI occurrences were associated with low air temperature, high relative humidity and high air pressure as well as with east & central migration routes of migratory birds. The average normalized difference vegetation index was 0.36 +/- 0.11 in epidemic areas of HPAI.

CONCLUSIONHPAI was unrandomly distributed and geographically clustered in China.

Animal Migration ; Animals ; Atmospheric Pressure ; Birds ; virology ; China ; epidemiology ; Cluster Analysis ; Environment ; Geographic Information Systems ; Humidity ; Influenza A Virus, H5N1 Subtype ; pathogenicity ; Influenza in Birds ; epidemiology ; Temperature

This paper reviewed the worldwide research progresses of the genus Laggera both on phytochemical and pharmacological work in the past few decades. The main secondary metabolites of this genus are proved to be sesquitepenoids, flavonoids and phenolic acids. Phamacological investigations revealed that the certain extracts of some Laggera species possess significant bioactivities on anti-inflammation, anti-tumor and anti-viral infection. This review afforded the comprehensive description of the active components as to provide useful references to elucidate their historical clinical application on upper respiratory infection, influenza, parotitis, and recurrent herpes viral infection.
Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; therapeutic use ; Antineoplastic Agents, Phytogenic ; pharmacology ; therapeutic use ; Antiviral Agents ; pharmacology ; therapeutic use ; Flavonoids ; chemistry ; isolation & purification ; therapeutic use ; Humans ; Influenza, Human ; drug therapy ; Molecular Structure ; Parotitis ; drug therapy ; Phytotherapy ; Plants, Medicinal ; chemistry ; Ranunculaceae ; chemistry ; Respiratory Tract Infections ; drug therapy ; Sesquiterpenes ; chemistry ; isolation & purification ; therapeutic use