The defense mechinism of heat shock protein(HSP)acting as moleculars chaperons requires heat shock transcription factors as the primary mediators of the heat shock response.Increased expression of HSP may protect the heart.Overexpression of HSP can inhibit cardiomyocyte apoptosis,protect the integrity of the microtubules and the actin cytoskeleton in cardiac myocytes and endothelial cells exposed to ischemia,participate in the folding and activation of protein kinases and transcription factors such as HIF-1? and HSF1,bind to endothelial nitric oxide synthase(Enos) and stimulate its activity,and downregulate cytokine production.Hence,it is of benefit to efficiently elevate HSP expression in myocardium using pharmacological and/or gene therapy procedures without any adverse effect.

Electrical status epilepticus during sleep(ESES)is a special electroencephalography in children.It is involved in a variety of epileptic syndromes and can lead to different degrees of degradation of movement, cognition, behavior, language.At present, the cause of ESES is still unknown.According to literature reports, 8 pathogenic genes have been found to be related to ESES, namely GRIN2A, CNKSR2, SCN2A, KCNA2, KCNQ2, KCNB1, SLC6A1 and WAC genes.In this review, progress of ESES molecular genetics are discussed.

OBJECTIVE:To establish a HPLC method for the determination of oxymatrine(OMT)and matrine(MT)in dog plasma.METHODS:Plasma protein was precipitated with perchloric acid;OMT and MT were extracted with dichloromethane under strong base condition and then plasma levels of OMT and MT were determined by HPLC.RESULTS:The linear ranges of OMT and MT were 0.2～ 15? g? mL-1 and 0.1～ 5? g? mL-1,respectively,and the detection limits of OMT and MT were 20ng? mL-1.The absolute recoveries,relative recoveries,intra-day and inter-day precisions of OMT and MT were all in line with the standards.CONCLUSION:The method is simple,accurate and sensitive yet with little interference,and it is applicable for the determination of OMT and MT in plasma,the studying of the pharmacokinetics,the transformation process and the metabolic pathway of OMT in vivo,furthermore,it serves as guidance for the development of new OMT preparations.

Objective To compare the results of manually -tested speech recognition threshold (SRT ) with automatically software -recorded SRT in the trial of Mandarin disyllabic test ,exploring the significance to the clini-cal applying .Methods 128 normal people of different ages without hearing loss and 57 workers exposed to noise in an automobile manufacturing was selected .These two group of volunteers speak mainly Mandarin in their daily life . MADSEN Conera (Danmark) clinical audiometr was applied .A group of double syllable word list with the same dif-ficulty of equivalence was used as test material .The initial presentation level was 20 dB HL higher than PTA .Then compared the results of manually -tested SRT with automatically software -recorded SRT .Results In the normal group ,the automatic value SRT was 7 .84 ± 3 .98 dB HL ,the manual value was 9 .19 ± 4 .47 dB HL ,and the average value of speech frequency threshold was 7 .63 ± 5 .78 dB HL .In the noise group ,the automatic value SRT was 6 .10 ± 8 .40 dB HL ,the manual value was 18 .81 ± 9 .52 dB HL ,and the average value of language frequency threshold was 27 .18 ± 19 .13 dB HL .There was significant difference between the values of SRT tested manually and recorded automatically (P<0 .01) .Conclusion There are differences between SRT valued manually and automatically .The SRT in people with normal hearing can be tested using automatic -recorded method .This method is convenient for screening in people without hearing loss .To exam in people with hearing loss ,the manual test is more appropriate .

To investigate the apoptotic effect of free fatty acids (FFAs) on human ovary granulosa cells, the granulosa cells were treated with FFAs in various concentrations for three days, and then the cell viability was determined by trypan blue exclusion method. DNA fragmentation was examined by DNA ladder formation and Annexin V GFP/PI staining of the cells. We also determined if the apoptotic effect of saturated FFAs was mediated by their acly CoA metabolites and measured the expression of apoptosis related genes, Bcl 2 and Bax by Western blot. The results indicated that saturated FFAs induced apoptosis of granulosa cell by their acly CoA metabolites in a dose dependent manner, and led to the down regulation of Bcl 2 and the up regulation of Bax. It is concluded that saturated FFAs can induce apoptosis of human ovary granulosa cells, suggesting that the sexual disorder of obese and insulin resistant women is related to it.

Objective:To investigate if chloride channel ClC-2 could be phosphorylated by mitogen activated protein kinase(MAPK) for further study of its regulation mechanism in proliferation and differentiation of cells.Methods:The coding sequence containing the cytosolic C-terminus of ClC-2 was amplified from pSPORT1/ClC-2 plasmid,including rabbit ClC-2 cDNA, by polymerase chain reaction(PCR),the fragment was cloned into pGEX-4T-1 plasmid for the construction of GST-tagged fusion protein expressing vector, pGEX-4T-1/ClC-2CT.After being identified by enzyme digestion and sequencing, the recombinant vector was transformed into a strain of E.coli BL21. The expression of GST-tagged fusion protein was induced with IPTG and purified with Gluthathion Sepharose 4B affinity chromatography. Then the phosphorylation of ClC-2 by MAPK was examined by using phosphorylation assays in vitro.Results:The construction of pGEX-4T-1/ClC-2CT recombinant vector was proved by enzyme digestion and sequencing. The purified fusion protein GST/ClC-2CT could be phosphorylated by MAPK, however the GST could not.Conclusion:Chloride channel ClC-2 can be phosphorylated by MAPK.

Objective To observe the impact of different culture conditions on the growth of the endothelial progenitor cells(EPCs) from rat peripheral blood. Methods Mononuclear cells obtained from rat peripheral blood were isolated by using density gradient centrifugation. According to the culture medium added with vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and the dishes were precoated with fibronectin or not, the mononuclear cells were divided into different groups and cultured under different conditions in vitro to compare the differences of the growth conditions. The different growth conditions were recorded and the result was calculated by statistics software. At last, the cells were identified by immunohistochemistry and immunofluorescence. Results The mononuclear cells from rat peripheral blood grew in the manner of keeping close to the wall in vitro. The cells and cell colonies cultured for 7 days hinted:under the same culture conditions, precoating FN was beneficial to the adherent proliferation of EPCs (t=4.43, P＜0.05; t=3.70, P＜0.05). Excluding the impact of fibronectin, growth factors could promote mononuclear cells differentiation to EPCs, indicating that growth factors enhanced proliferation of EPCs (t =-13.22, P＜0.01; t =-10.96, P＜0.01). Immunohistochemistry and immunofluorescence showed that, at day 4, 7, 10 of cells cultured, CD34 and CD133 expressions were increased gradually [(35.7±4.2)%, (60.1±3.8)%, (81.8±6.4)%; (3.2±0.9)%, (18.4±7.3)%,(32±3.8)%, respectively]; at day 14, both were decreased [(32.1±5.4)%, (1.9±2.7)%]; but Flk-1was increased at day 4, 7,10, 14 [(31.2±3.5)%, (40.6±5.3)%, (71.2±8.4)%, (81.5±4.1)%].Conclusions Fibronectin is conducive to adhesion and proliferation of EPCs. VEGF and bFGF play an important role in the differentiation of EPCs. The success culture of EPCs in vitro will provide a sufficient number of seed cells for its application in vascular tissue engineering, and offer new ideas for peripheral blood stem cell transplantation for the treatment of various diseases.

Objective: To establish a scoring model for liver cirrhosis disease (SLCD) in experimental rats, so as to provide evidence for early clinical prevention and treatment of liver cirrhosis diseases. Methods: The liver cirrhosis model was induced in rats by composite factor method. The damages of hepatic tissues and the changes of serological liver function were observed in different phases of the rat liver cirrhosis model. The SLCD formula was obtained using the Master-Element Analysis method. Results: The experimental liver cirrhosis model was successfully established in rats. The experimental rat SLCD and SLCD formula were successfully constructed using the following 6 parameters, the age, total bilirubin, albumin, prealbumin, prothrombin time-international normalized ratio, and serum creatinine. The score was expressed as the R value, which gradually decreased with the aggravation of lesion and fibrosis. R=1: the liver tissue was normal; 0.702 ≤ R < 1: there were inflammatory reaction, focal degeneration and necrosis in the liver; 0.542 ≤ R < 0.702: there was fibroplasia in the liver; 0.352 ≤ R < 0.542: pseudolobules were formed, and it was in the liver cirrhosis stage; and R<0.352: it was in the later stage of liver cirrhosis. Conclusion: The SLCD model can sensitively and accurately display the liver impairment and liver function reserve during experimental liver cirrhosis in rats. It may help the early prevention, diagnosis and treatment of the cirrhosis diseases if relevant clinical evidence is obtained.

Objective To investigate the effect of hydroxysafflor yellow A (HSYA) on neurogenesis of the dentate gyrus organotypic hippocampal slice cultures after oxygen-glucose deprivation (OGD).Methods The 3-day-old Sprague Dawley rats were used to making organotypic hippocampal slices.They were randomly divided into four groups:a blank control group,a saline control group (giving 1 mL normal saline after OGD),a high-dose HSYA group (giving 1 ml,0.072 mg/ml HSYA after OGD),and a low-dose HSYA group (giving 1 ml,0.036 mg/ml HSYA after OGD) (n =6 slices in each group).Nestin and 5-bromo deoxyuridine nucleoside imnune fluorescence method were used to double-label the proliferation of neural stem cell (NSC).Results There were significant differences in NSC numbers among the different time points before and after OGD in slices of each group (all P ＜0.001).After OGD,the proliferation of NSC numbers reduced significantly in each group,cell proliferation increased siginificantly at day 3,and the proliferation of NSC number at day 6 was lower than that at day 3,but they still higher than those before OGD.There were significant differences in the proliferation of NSC numbers among the blank control,the saline control,high-dose HSYA,and low-dose HSYA groups (F =947.077,P ＜0.001).Observing from each time point,in addition to before OGD,the proliferation of NSC numbers at all other time points had the following relations:the blank control Group ＞ the high-dose HSYA group ＞ the low-dose HSYA group ＞ the saline control group (all P＜0.001).Conclusions HSYA may alleviate OGD to the NSC injury and contribute to neurogenesis.

OBJECTIVE:To explore the clinical role of two-hour(C2)of CsA in allogeneic hematopoietic stem cell transplantation.METHODS:The whole blood CsA concentration of 9 patients of C0 and 12 of C2 were determined by FPIA method . RESULTS: The satisfactory immunosuppressive concentration was found as follow: C0 was 200～400 ?g?L-1and C2 was 500～700 ?g?L-1.Both C0 and C2 could predict the acute rejection and side-effects,but C2 is more effective.CONCLUSION:The factors that affect the whole blood concentration of CsA were complex. CsA blood concentration monitoring can be used to guide rational use of CsA,but C2 is more effective than C0.