Objective To explore the pathogenic mechanisms of intractable epilepsy by searching for the differential cell signal transduction associated genes expression in intractable and non intractable epilepsy rat brain using cDNA microarray Methods Intractable epilepsy and non intractable epilepsy rat model were build The total RNAs were isolated from the brain tissues Both mRNAs from the brains of the intractable and non intractable epilepsy rats were reversely transcribed to the cDNA with the incorporation of fluorescent dUTP to prepare the hybridization probes The PCR products of 4 096 human genes were spotted on a chemical material coated glass plates in array The mixed probes were then hybridized to the cDNA microarray After high stringent washing, the cDNA microarray was scanned for the fluorescent signals and showed differences between 2 tissues Results Among the 4 096 target genes, 29 genes associated with cell signal transduction differentially expressed were identified, 10 were up regulated(34 48%) and 19 down regulated(65 52%) Conclusion cDNA microarray technology is an effective technique in screening the differentially expressed genes between intractable and non intractable epilepsy rat brain Disturbances of cell signal transduction play a role in the pathogenic mechanism of intractable epilepsy

Objective To investigate the expression of microRNA-134 ( miR-134 ) , CREB and pCREB in the temporal lobe tissue of patients and epileptic rats and to explore their roles in pathogenesis of epilepsy.Methods Tempo-ral lobe tissue samples of 14 patients with refractory epilepsy and 10 non-epileptic patients, and hippocampus and brain tis-sue samples of 42 rats were used in this study.Forty-two healthy adult male Sprague-Dawley rats were randomly divided in-to 6 epilepsy groups (24 h, 72 h, 7 d, 14 d, 30 d, and 60 d after kindling epilepsy) and a normal control group (n=6 for all groups) .The rat model of epilepsy was generated by intraperitoneal injection of 127 mg/kg lithium chloride and 16-20 h later, 35 mg/kg pilocarpine.In the temporal lobe tissue of patients and hippocampal tissue of rats, the expression level of miR-134 was detected by real-time polymerase chain reaction.The expression levels of CREB and pCREB were de-termined by Western blot, and CREB and pCREB localization was assessed by immunohistochemistry.Results Compared with the control rats, the expression of miR-134 was significantly decreased in the temporal lobe tissue of experimental rats at 72 h,7 d,14 d, 60 d after kindling (P＜0.05),and no significant change at 24 h and 30 d after kindling (P>0.05). Expression of miR-134 in patients with refractory epilepsy was significantly lower than that of the controls ( P＜0.05 ) , while up-regulation of CREB expression was at the same time points (P＜0.05).Up-regulation of pCREB expression was at all the time points after kindling (P＜0.05).CREB and p-CREB expressions were seen in the nuclei of neurons, and significantly higher in patients with refractory epilepsy and epileptic rats.Conclusions The expression of miR-134 is sig-nificantly decreased and that of CREB and pCREB was significantly increased in the temporal lobe tissue of patients with re-fractory epilepsy and the hippocampal tissue of epileptic rats.These findings indicate that the signaling pathway of miR-134/CREB/pCREB may play an important role in the pathogenesis of epilepsy.

Objective: To investigate the anti-inflammatory, bacteriostatic, diuretic, antispasmodic and immunoenhancement effect of Niao Gan Ning(NGN). Methods: The tests of inhibiting the leukocyte infiltration, in-vitro bacteriostasis, water loading with metabolism cage, isometric tension recording and the phagocytosis of abdominal macrophages(AM) were conducted. Results: NGN could obviously inhibit the wandering of leukocyte in abdominal cavity and decreased the number of WBC in mice, and exert bacteriostatic effect to various degrees. NGN also increase the urine volume in water load rats, inhibited contraction of bladder and ureter smooth muscle induced by Carbacol and histamine in rabbits and rats, and increased phagocytosis of the chicken red cells by AM. Conclusion: NGN has a strong anti-inflammatory, bacteriostatic, diuretic and antispasmodic effect.

OBJECTIVETo investigate the expression and activity of silent information regulator 1 (SIRT1) in the temporal lobe of epileptic patients and rat models and explore its role in the occurrence and progression of epilepsy.

METHODSThe temporal lobe tissue of epileptic patients and rat models (induced by lithium-pilocarpine) were examined for SIRT1 expression using immunohistochemistry and Western blotting and also for SIRT1 activity using SIRT1 Deacetylase Assay Kit.

RESULTSImmunohistochemistry detected positive SIRT1 expression mainly in the cytoplasm of the neurons in both human and rat brains, and the epileptic groups showed stronger SIRT1 immunoreactivity than the control group. Western blotting and activity assay showed that the expression and activity of SIRT1 were significantly increased in the temporal lobe of patients with refractory epilepsy as compared with the tissues samples from non-epileptic patients (P<0.05). In the rat models of epilepsy, SIRT1 expression was up-regulated at 6, 24, and 72 h and at 7, 14, 30, and 60 days after kindling (P<0.05) and SIRT1 activity was significantly increased at 6, 24, and 72 h and at 7 and 14 days (P<0.05), with the peak level of SIRT1 expression and activity occurring at 72 h.

CONCLUSIONUp-regulation of SIRT1 expression and activity in the temporal lobe of epileptic patients and rat models may play an important role in the pathogenesis of epilepsy.

Adolescent ; Adult ; Animals ; Disease Models, Animal ; Epilepsy ; metabolism ; Female ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Sirtuin 1 ; metabolism ; Temporal Lobe ; metabolism ; Young Adult

When we try to establish the gene recombinant yeast cell to screen the androgenic endocrine disruptors, the key procedure is the androgen receptor (AR) expression in the yeast cell. For this purpose, we obtained the GPD (glyceraldehyde-3-phosphote dehydrogenase) promoter from the yeast genosome of W303-1A using PCR system and inserting it into Swa I and BamH I sites of pYestrp2. The new constructed vector was named pGPD. The V5 epitope tag DNA with a 5'-BamH I and a 3'-EcoR I sticky end was cloned into the corresponding site of the pGPD vector to yield the vector of pGPDV5. The 2 723 bp full length AR ORF amplified by PCR from pcDNA3.1/AR was fused to V5 epitope tag DNA in pGPDV5 to give the AR yeast expression vector of pGPDV5/AR. This fused vector was transformed into the yeast cell (W303-1A). Western blot was used to detect the V5 fused protein of AR, in the protocol of which the primary monoclonal antibody (IgG(2a)) of mouse anti-V5 and the polyclonal secondary antibody of goat anti-mouse (IgG) linked to horseradish peroxidase (HRP) were used to detect the specific protein in the given sample of the transformed yeast extract. The result showed that the fused protein of AR was expressed successfully in the yeast cell.
Base Sequence ; Endocrine Disruptors ; analysis ; Epitopes ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Glyceraldehyde-3-Phosphate Dehydrogenases ; genetics ; Humans ; Molecular Sequence Data ; Promoter Regions, Genetic ; Receptors, Androgen ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; genetics ; Yeasts ; genetics ; metabolism

OBJECTIVETo explore the relationship between the syndrome types in traditional Chinese medicine (TCM) and serum HBV DNA load in chronic HBV carriers positive for HBeAg.

METHODSAccording to the TCM syndrome types, 185 chronic HBV carriers with HBeAg positivity were classified into single syndrome group (liver Qi depression, kidney Qi deficiency, spleen Qi deficiency, and kidney Yang deficiency), compound syndrome group, and unidentifiable syndrome group; based on the nature of the condition in TCM terms, the patients were classified into excess syndrome group, deficiency syndrome group and comorbidity syndrome group. The serum HBV DNA levels in these cases were analyzed in relation to the TCM syndrome types and disease nature.

RESULTSHBV DNA levels showed no significant difference among the patients with single syndrome, compound syndromes and unidentifiable syndrome (F=0.910, P=0.404), nor among the patients with the 5 different single TCM syndromes (χ²=4.672, P=0.323) or those with different disease nature (F=0.631, P=0.596).

CONCLUSIONSerum HBV DNA level can not be considered as the evidence for syndrome differentiation in chronic HBV carriers with positive HBeAg.

Adolescent ; Adult ; Carrier State ; blood ; diagnosis ; Child ; DNA, Viral ; blood ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; blood ; diagnosis ; virology ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Young Adult