DNA methylation in the medial prefrontal cortex regulates alcohol-related behavior in rats.

Alcohol abuse causes tissue and organ damage, and may participate neuropsychiatric diseases. Studies have shown that DNA methylation plays an important role in gene expression and behavioral changes induced by alcohol, however the causative neurobiological mechanisms have not been clarified. In this study, 32 healthy adult male SD rats were randomly divided into a drinking water control group (n=16) and a chronic alcohol exposure group (n=16). The alcohol preference and locomotor activity of rats were evaluated by two-bottle choice test (TBCT) and open-field test (OFT). DNA methylation in the medial prefrontal cortex (mPFC) tissue was detected by the reduced representative bisulfite sequencing (RRBS) technology. The methylation differential genes closely related to alcohol abuse were screened. qRT-PCR was used to verify the mRNA expression patterns of differential genes. qRT-PCR and Western blot were used to detect the expression of DNA methyltransferases (DNMTs) and methyl CpG binding protein 2 (MeCP2). Furthermore, the effect of short-term alcohol exposure (7 days) on DNMTs and MeCP2 in the mPFC of rats was tested (n=8/group). The results indicated that the methylation level of promoter region in the mPFC of rats exposed to chronic alcohol was significantly increased. In addition, the increased methylation levels in the promoter of Ntf3 and Ppm1G were accompanied by down-regulated mRNA levels in the chronic alcohol exposure group. The decreased methylation levels in the promoter of Hap1 and DUSP1 were accompanied by up-regulated mRNA levels. Furthermore, chronic alcohol exposure increased the mRNA and protein levels of DNMT3B and MeCP2. However, short term alcohol exposure did not affect their expression. This present study provides evidence that DNA methylation is associated with the development of alcohol abuse, which may be regulated by DNMT3B and MeCP2. The target genes Ntf3, Ppm1G, Hap1, and DUSP1 related to alcohol abuse were discovered as well, providing new insights into the neurobiological mechanism of alcohol abuse and the potential pharmacological targets for the treatment of alcohol abuse.

[Survey on host animal and molecular epidemiology of hantavirus in Chuxiong prefecture, Yunnan province].

OBJECTIVE: To determine the hosts of hantavirus (HV) and its molecular epidemiological characteristics, to provide evidence for prevention and control on hemorrhagic fever with renal syndrome (HFRS). METHODS: Rodents were captured by a special trap within the residential area. The antigens of HV in lung tissues were detected by direct immuno-fluorescence assay (DFA). Nucleotide sequences of HV were amplified by RT-PCR with HV genotype-specific primer. The amplified genes were then sequenced. Phylogenetic tree were built on nucleotide sequence with ClustalX 1.83 software. RESULTS: 1421 rodents were captured and classified into 8 species of 4 Genera in the epidemic area within 10 counties of Chuxiong prefecture, Yunnan province, between 2005 and 2006. Out of the 1421 rodents, 1056 (74.31% ) of them were Rattus norvegicus and 280 (19.70%) belonged to Rattus flavipectus. The antigens of HV were detected by DFA in lung tissues and the total positive rate of HV was 5.15% (53/ 1029). After applying the sequencing nucleotide method to the 53 positive specimens, data showed that 21 specimens were positive and all of them belonged to Seoul type (15 samples were from Rattus norvegicus, 4 samples Rattus flavipectus, 2 samples Rattus nitidus). The partial S segments from 12 specimens were sequenced which appeared homologic with R22, L99 and HLD65 from GenBank in relatively high level (87.1% -99.7%). When compared to 76-118 strain of Hantaan type, their homologic degree was only 64.4%-69.1%. Results from Phylogenetic analysis showed that 12 specimens belonged to Seoul type. As for their homology, they were significantly similar to Seoul type and could be tentatively divided into two subtypes S1 and S3. CONCLUSION: It was confirmed that the Seoul type virus, as HFRS's pathogenetic agent mainly carried by rats, prevailed widely in Chuxiong prefecture. Owing to the local ecological environment, we also noticed the characteristics of different HV subtypes among Seoul type.

[Survey on human rabies cases and its viral molecular biological features in Baoshan city, Yunnan province].

OBJECTIVE: To understand the epidemiological features of two rabies cases in Baoshan city year 2006 and 2007 and to analyze its source of infection. METHODS: Questionnaires were used to do the epidemiological survey on each of the rabies cases. Brain tissue samples of rabies patients were collect to detect the rabies virus by direct immunofluorescence assay (DFA) and RT-PCR assay. Homology and phylogenetic tree were analyzed, based on the whole nucleotide and deduced amino acid sequence of P, M and N gene of rabies virus followed by molecular epidemiological analysis. RESULTS: In July 2006, one human rabies case was identified in Longyang district, and another one in Tengchong county in Baoshan city in 2007. The degrees of exposure of these two patients was all at degree III. Two brain tissue samples among the dead patients (No. CYN0601H and CYN0701H) were confirmed positive by both DFA and RT-PCR assay. The homology analysis of P, M and N gene sequences among CYN0601H, CYN0701H and other rabies strains isolated from other provinces and other counties, showed that the samples in Baoshan city shared the highest homology with the strains in Thailand. Phylogenetic analysis indicated that the two samples were very close and all belonged to genetype 1 Lyssavirus, with the closest relationship between samples in Baoshan city and strains in Thailand. CONCLUSION: It was confirmed on the virus molecular level that the two patients in Baoshan city were both suffered from rabies. The prevalent strains in Baoshan city was probably imported from foreign country, suggesting that prevention and control measures on rabies virus in the boarder areas of Yunnan should be strengthened.

Effect of Kaixin Capsule in Reconstructing the Collagen of Non-infarct Region of Left Ventricle in Rats after Myocardial Infarction / 广州中医药大学学报

To explore the effect of Kaixin Capsule (KC, composed of Radix Panacis Quinquefolii, Radix Astragali, Radix Ophiopogonis, Rhizoma Chuanxiong,Rhizoma Cyperi, etc.) in reconstructing the collagen of non infarct region of left ventricle in rats after myocardial infarction (MI) and to study its mechanism.Seventy rats were allocated to mimic operation group (Group Ⅰ), model group (Group Ⅱ), high dosage KC group (Group Ⅲ), low dosage KC group (Group Ⅳ) and positive control group (Group Ⅴ). Rat models of left ventricle reconstruction were established by the ligation of the main stem of left coronary artery. Three days after modeling, the rats were treated with KC for eight weeks. And then the total collagen content in the myocardium of non infarct region and angiotensin Ⅱ (AngⅡ) and aldosterone (ALD) levels in blood and myocardium were measured by oxyproline hydroxyproline method and radioimmunoassay. The total collagen content was higher in Group Ⅱ than that in Group Ⅰ (P0 05) . [Conclusion]KC can inhibit the production of collagen and increase myocardial compliance and its mechanism may be involved in the regulation of rennin angiotensin aldosterone system.