A nest RT-PCR/restriction test has been developed in order to distinguish the lapinised vaccine strain from field isolates of classical swine fever virus. The restriction enzyme cut sites mapping of the major coding sequence of E2 gene lapinised vaccine strain and ShiMen strain of classical swine fever virus have been compared. Ten and sixteen unique restriction markers have been found in the lapinised vaccine strain and ShiMen strain. The restriction enzyme cut sites mapping of the twenty six unique restriction marker in the major coding sequence of E2 gene of 17 classical swine fever field isolates have been analyzed. Only 3 sites (HgaI、Hin8I及Hsp92I) are present in the lapinised vaccine strain sequence. Two pans of nested primers and a criteria of analysis have been designed for HgaI restriction marker site. The tests have been conducted first on the lapinised vaccine strain and ShiMen strain of classical swine fever virus resulting in predicted restrection patterns. Finally, the tests have been applied to 5 field isolates of different gene group analyzed by phylogenetic study. The result showed that only HCLV strain gene can be cut to 2 fragment by Hgal , and ShiMen strain and 5 field isolates cant be cut At the same time the sensitivity and specificity of nest RT-PCR have been tested. The sensitivity is 0. 2MLD. The specific fragment of BDV and BVDV were not obtained by the nest RT-PCR. These results showed that the development of the nest RT-PCR/restriction tests is very important for the control and perish of classical swine fever in china.

Acute Disease ; Humans ; Insecticides ; poisoning ; Male ; Occupational Exposure ; adverse effects ; Poisoning ; etiology ; prevention & control

OBJECTIVETo establish a hepatolenticular degeneration rat model with excessive copper, and investigate the effects of excessive copper deposits in the liver on hepatocyte apoptosis and Bax and Bcl-2 expression.

METHODSRat model of hepatolenticular degeneration was established by administering forages containing 1g/kg of copper sulfate and drinking water containing 0.185% copper sulfate. Copper level in the liver and serum and alanine aminotransferase (ALT) level in serum were measured using an atomic absorption spectrophotometer. The terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate nick-end labeling (TUNEL) method was used to detect hepatocyte apoptosis. Bax and Bcl-2 expression was observed by RT-PCR and imunohistochemistry staining. Quantification of positive cells was performed by image analyzer.

RESULTSWith more prolonged excessive copper ingestion, copper level in the liver and serum as well as ALT level in serum rose, and more apoptosis cells appeared in the liver. Bax and Bcl-2 expression increased significantly compared with controls fed a normal diet and progressively increased with more prolonged excessive copper ingestion. The progressively increased extent of Bcl-2 expression was lower than that of Bax expression, so the ratio of Bcl-2/Bax decreased with increasing excessive copper ingestion time.

CONCLUSIONSExcessive copper deposits in the liver can induce hepatocyte apoptosis through an up-regulation of Bax expression.

Alanine Transaminase ; blood ; Animals ; Apoptosis ; drug effects ; Copper ; toxicity ; Hepatocytes ; drug effects ; pathology ; Liver ; chemistry ; metabolism ; Male ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; bcl-2-Associated X Protein ; analysis ; genetics

There are increasing reports of methanol poisoning (MP) incidence worldwide. In Malaysia, the largest first methanol poisoning was reported in Selangor in 2013 with a total of 41 patients and cluster of cases been reported from the country since then. Often MP involved adulterated alcohol containing more than the legal permissible concentration of methanol. Methanol is rapidly absorbed and metabolised into formic acid which causes variable symptoms of the central nervous system such as blindness, seizure, coma and gastrointestinal disturbances. Mortality could reach up to 83% as reported using the coma state, pH and pCO2 level in the worst-case scenario.