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

Brain Stem/pathology* ; Death, Sudden/etiology* ; Fatal Outcome ; Humans ; Intracranial Hemorrhages/mortality* ; Leukemia/pathology*

To develop a real-time FQ-PCR method for quantifying human ermap, a set of primers and a fluorescent probe were designed by primer express 2.0. pBluescriptSK(+) plasmid contained ermap cDNA was transcribed to generate calibration standards for quantification. A real time FQ-PCR method was established. The results showed that when the concentrations of DNA to be amplified were ranged from 1.725 x 10(7) to 1.725 x 10(10) cps/ml, there was a good correlation between template concentration and cycle threshold, and the correlation coefficient reached to -0.999376. In conclusion, real time FQ-PCR which is specific, sensitive and accurate can be used to further research on human ermap.
Blood Group Antigens ; genetics ; Butyrophilins ; DNA, Complementary ; chemistry ; genetics ; Fluorescent Dyes ; chemistry ; Fluorometry ; methods ; Humans ; Polymerase Chain Reaction ; methods ; Reproducibility of Results

AIMTo study the pharmacokinetics of genistein in Beagle dogs.

METHODSGenistein, suspended in 0.5% CMC-Na solution, was orally administered to Beagle dogs at the dose of 5.34 mg.kg-1. At various time intervals, 1.5 mL of blood was drawn from the vein of dogs in their front legs. At the same time, urine and feces were collected. After the collection, the feces were homogenized with physiological saline (to 1 g feces, 10 mL physiological saline were added). The genistein in plasma, urine and homogenized feces was extracted twice by vortexing with 2.0 mL mixture of methyl tert-butyl ether and pentane (8:2). The organic phase was transferred into tubes and evaporated in ventilation cabinet. The residue was dissolved in 50 microL of methanol and 20 microL of the solution was drawn and detected by high-performance liquid chromatography. The pharmacokinetic parameter was calculated by 3P97 software.

RESULTSThe plasma concentration-time curve was fitted to a one-open-compartment model. The peak time was 0.29 h, and the elimination half-life was 0.52 h. After genistein was administered, 10.79% of genistein were excreted from urine and 21.55% from feces within 24 h. It was also found that 13.00% genistein were excreted from urine and 52.46% from feces within 60 h.

CONCLUSIONIt showed that the speed of absorption and elimination of genistein was high in Beagle dog, and genistein was mainly excreted in the form of parent compound in urine and feces.

Animals ; Anticarcinogenic Agents ; blood ; pharmacokinetics ; urine ; Area Under Curve ; Chromatography, High Pressure Liquid ; Dogs ; Feces ; chemistry ; Genistein ; blood ; pharmacokinetics ; urine

OBJECTIVETo observe the pathological and clinical characters of nonalcoholic steatohepatitis (NASH).

METHODSLiver biopsy tissues taken from 97 patients negative for common viral detection with serological test and immunohistochemistry as well as in situ hybridization, were observed by routine light microscopic examination. And the clinical data of patients with NASH was analyzed.

RESULTSAmong the chronic hepatitis patients with unknown etiology, the detection rate of NASH was 15.5% (15/97). The characteristic lesions in NASH patients included macrovesicular steatosis in zone 3 of lobules, hepatocytes ballooning degeneration, lobules diffused with acute and chronic inflammation, and perisinusoidal fibrosis. Grading and staging according to Brunt's method, histological lesions could be accounted for G1S1 in 7 patients, G2S2 in 3 patients, G3S1 in 4 patients and G3S2 in 1 patient. There were 14 patients with mild to moderate elevation of transaminase, 10 with hyperlipemia, 8 with diabetes and 9 with fatty liver by ultrasonography.

CONCLUSIONNonalcoholic steatohepatitis is a common form of unknown etiology chronic liver disease with certain clinic-pathological features.

Adult ; Biopsy ; Fatty Liver ; pathology ; Female ; Humans ; Liver ; pathology ; Male ; Middle Aged

OBJECTIVETo compare the value of apolipoprotein B (apoB) and low-density lipoprotein cholesterol (LDL-C) in assessing the risk of coronary heart disease in patients with inconsistent apoB and LDL-C levels.

METHODSIn a total of 603 patients undergoing coronary angiography, apoB and LDL-C levels were categorized into high and low levels relative to the median levels of apoB and LDL-C, based on which the patients were divided into 4 groups with low apoB/low LDL-C, low apoB/high LDL-C, high apoB/low LDL-C, or high apoB/high LDL-C. According to the results of coronary angiography, we evaluated the number of coronary artery branches with lesions and the severity of coronary artery stenosis in the 4 groups to assess the correlation of apoB and LDL-C with cardiovascular risks.

RESULTSWe found significant differences in the number of coronary artery branches with lesions and the severity of coronary artery stenosis among the 4 groups (P<0.05). The number of coronary artery branches involved and the severity of stenosis differed significantly between patients with consistently high and low apoB/LDL-C levels (P<0.005). Compared with those with low apoB/low LDL-C levels, the patients with high apoB/low LDL-C levels showed a significantly greater number of coronary artery branches with lesions (P=0.017) and more severe stenosis (P=0.034), but such differences were not found in patients with low apoB/high LDL-C levels. Pearson correlation analysis identified LDL-C and apoB as the risk factors for cardiovascular disease with areas under the ROC curve of 0.579 (P=0.014) and 0.589 (P=0.006), respectively.

CONCLUSIONSIn patients with inconsistent levels of apoB and LDL-C, apoB and LDL-C levels are both risk factors of coronary heart disease in close relation with the disease severity. LDL-C and apoB are comparable for their important values in predicting the risk of coronary heart disease.

Objective To understand the spatial and temporal distribution characteristics of dengue fever in China from 2011 to 2018, and predict the incidence of dengue fever in China in 2019. Methods Based on the case data of dengue fever in China from 2011 to 2018 in the Chinese Disease Prevention and Control Information System, the trend of dengue fever was described and predicted by using the autoregressive integrated moving average model (ARIMA) with R 3.6.0 software. Based on the data of the incidence of dengue fever in the country, provinces and cities from 2011 to 2016 provided by the national scientific data sharing platform for population and health, global and local spatial autocorrelation analysis was performed using GeoDa 1.12 software to determine the dengue fever hotspots. Results The incidence of dengue fever was 14 302 in 2019, showing no disease outbreaks. The incidence of dengue fever in 2012(Moran’s I=-0.088, P=0.037), 2013(Moran’s I=-0.121, P=0.040) and 2014(Moran’s I=-0.076, P=0.045) showed a global spatial negatively correlaton. In 2016(Moran’s I=0.078, P=0.048), the incidence of dengue fever was positively correlated with global space. The results of local autocorrelation analysis showed that the high incidence of dengue fever was mainly in the southeast coastal areas of China. Conclusions In 2019, the epidemic of dengue fever in China showed no obvious fluctuation trend, and the epidemic situation showed spatial clustering distribution.

Purpose: The purpose of this study was to investigate the prevalence and risk factors of overactive bladder (OAB) in young adults and to explore the influence of OAB on mental health. Methods: Between October 2019 and January 2020, 14,010 anonymous questionnaires were distributed to freshmen at 2 universities in Henan, China. The students came from all over the country. The questionnaire included general items and information necessary to calculate the overactive bladder symptom score, the Chinese version of the Pittsburgh Sleep Quality Index (PSQI) score, Self-Esteem Scale (SES) score, and Self-Rating Depression Scale (SDS) score. The relationships between the prevalence of OAB and its risk factors were evaluated. Results: The overall prevalence of OAB was 6.0%, with 4.3% of participants characterized as having dry OAB and 1.7% as having wet OAB. The prevalence of mild OAB was 5.5%, and that of moderate OAB was 0.5%; no severe OAB was observed. Higher prevalence rates of OAB were found among women, respondents with constipation, and respondents with primary nocturnal enuresis (PNE) (P <0.05). Compared to healthy controls, the OAB group exhibited a higher mean SDS score (52.12±8.986 vs. 47.71±9.399, P<0.001) and mean PSQI score (5.28±2.486 vs. 4.27±2.431, P<0.001), but a lower mean SES score (27.78±3.599 vs. 29.57±4.109, P<0.001). Conclusions OAB significantly affects the mental health of young adults. Female sex, constipation, and PNE are risk factors for OAB.

A rice pse(t) (premature senescence, tentatively) mutant line, was isolated from 4,500 independent T-DNA inserted transgenic lines. The symptoms of premature senescence appeared more severely than those of the control plants (Zhonghua 11, japonica) at the last development stage. To characterize the mutant and provide basic information on the candidate genes by mapping to a physical region of 220-kb, experiments were carried out in two phytotrons under controlled temperature of 24 degrees C and 28 degrees C, respectively. The content of chlorophyll, soluble protein and MDA (malondialdehyde), net photosynthesis, the antioxidant enzyme activities of SOD (superoxide dismuase) (EC 1.15.1.1) and POD (peroxidase) (EC 1.11.1.7) and the peptidase activities of leaves were measured from top to bottom according to the leaf positions at the flowering stage. Compared with the control plant, the mutant showed the following characteristics: (1) Higher net photosynthesis rate (P(n)) appeared in the 1st and 2nd leaves, contents of chlorophyll and soluble protein were also higher in the 1st leaf; (2) The activities of SOD, POD and peptidase were higher according to the leaf position from top to bottom; (3) The symptom of premature senescence was accelerated in the mutant at 28 degrees C treatment. The MDA content and the SOD and POD activities between the 24 degrees C and 28 degrees C treatment mutants were not significantly different. Content of chlorophyll and soluble protein of leaves mutant decreased rapidly at 28 degrees C treatment. The results show that pse(t) is sensitive to high temperature. The probable function of PSE(T) is discussed.
Aging ; physiology ; Antioxidants ; metabolism ; Apoptosis ; physiology ; Mutation ; Oryza ; classification ; genetics ; growth & development ; Photosynthesis ; physiology ; Plant Leaves ; classification ; genetics ; growth & development ; Plant Proteins ; genetics ; metabolism

BACKGROUND: Primary injuries of the spinal cord are irreversible in the pathoplhysiological process. Most of studies have focused on prevention against secondary injuries, by reducing neuronal apoptosis and necrosis as well as decreasing damage area, which provides favorable environment for axonal regeneration, blocks secondary injury and promotes neural regeneration after spinal cord injury.OBJECTIVE: To review the recent advances in the signaling pathway related to secondary injuries after spinal cord injury,and to clarify the related signaling pathways, thereby providing theoretical basis for the treatment of spinal cord injury.METHODS: PubMed, EMBASE, Medline, CNKI, CJFD and Wanfang databases were retrieved for the articles addressing the signaling pathways in spinal cord injury published between 2006 to 2016. The keywords were "spinal cord injury, signaling pathway" in English and Chinese, respectively. The signaling pathways in secondary injuries after spinal cord injury were summarized.RESULTS AND CONCLUSION: Mitogen activated protein kinase, nuclear factor of kappa B, PI3K/PKB (Akt), Janus kinase/signal transducer and activator of transcription, and Wnt signaling pathways are mainly involved in secondary injuries after spinal cord injury. All these pathways play a significant role in the development and repair of spinal cord injury, and future investigation on them is warranted.