Based on M9 culture medium,the concentration of ingredients of culture medium was optimized for the fermentation of pNSR32/BL21(DE3),the engineering bacterial with spider silk protein,and lactose as an inducer.The condition of optimum culture medium was obtained for the expression of the high molecular weight recombinant spider silk protein by using orthogonal and individual factor experimental design.The result was showed that the optimum culture medium was consisted of 0.3% glycerol,3% yeast,0.75% tryptone,0.05%(NH_4)_2SO_4 and a little inorganic salt_.It was confirmed that the optimum culture medium will benefit the growth of bacterial and expression of recombinant spider silk protein.The production level of propose protein will attain at 20% of the total proteins in the fermentation.

OBJECTIVE:To investigate the effects of Wenxin granule on the related indexes of senile patients with primary hy-pertension and acute left cardiac failure. METHODS:160 senile patients with primary hypertension and acute left cardiac failure were randomly divided into control group(80 cases)and observation group(80 cases). Control group received 12.5 mg captopril, orally,twice to 3 times a day (if patients had taken plenty of diuretic drugs,in low sodium and/or low blood volume,patients with normal or low blood pressure,the initial dose was 6.25 mg,3 times a day). Observation group additionally given 9 g of Wenxin granule,3 times a day. The treatment course for both groups was 4 weeks. SBP,DBP,PP,SDANN,SDNN-Index,RMS-SD,PNN50,improvement of cardiac functions before and after treatment,and incidence of adverse reactions in 2 groups were ob-served. RESULTS:After treatment,the SBP,DBP and PP in 2 groups were significantly lower than before,and observation group was lower than control group,SDNN,SDANN,SDNN-Index、RMSSD and PNN 50 were significantly higher than before,and ob-servation group was higher than control group,the differences were statistically significant(P<0.05). And there was no significant difference in the incidence of adverse reactions(P>0.05). CONCLUSIONS:Based on conventional treatment,wenxin granule can significantly reduce the bloop pressure of senile patients with primary hypertension and acute left cardiac failure,improve cardiac functions and inhibit autonomic disorders,with good safety.

Objective:To explore the diagnostic value and correlation of serum tumor markers combined detection of pancreatic cancer. Methods:Selected January 2013 to may 2016 in our hospital in patients with pancreatic cancer in 146 cases,128 cases of non pancreatic cancer with patients and 124 cases of healthy physical examination. Radiation immunity analyzer test groups of serum CA19-9,CA242,CA50,CA125,CEA and TSGF levels and compared between groups. To draw the working characteristic curve(ROC) to analyze the diagnostic value of tumor markers in patients with pancreatic cancer and the correlation of linear correlation analysis of tumor markers. Analysis of independent risk factors for pancreatic cancer using multiple logistic regression models. Results: The pancreatic cancer group of serum CA19-9,CA242,CA50,CA125,CEA and TSGF levels were significantly higher than the control group and non pancreatic cancer group,the differences were statistically significant(P<0. 05 or P<0. 01).Ⅳstage andⅢstage with patients of serum CA19-9,CA242,CA50,CA125 and TSGF levels were significantly higher than Ⅰstage and Ⅱstage with patients(P<0. 01).Ⅳ stage with patients of serum CA19-9, CA242, CA125 and CEA levels were significantly higher than Ⅲ stage with patients ( P<0. 01). The pancreatic cancer group of serum CA19-9,CA242,CA50,CA125,CEA and TSGF of positive rate were significantly higher than that of control group and non pancreatic cancer group(P<0. 01). ROC curves showed that the AUC of serum CA19-9 were higher than other single indexes,the best critical value,sensitivity and specificity were 114. 5 U/ml,81. 2% and 79. 3% respectively. The diagnostic efficacy of the six joint detection were better than the single detection,the sensitivity and specificity were 92. 4% and 76. 5%respectively. Correlation analysis showed that serum CA19-9 were positively correlated with CA242,CA50 and CA125(r=0. 703,P=0. 005;r=0. 572,P=0. 024;r=0. 439,P=0. 036). Multiple logistic regression analysis showed that smoking,incorrect diet,history of diabetes,gallbladder disease history and high levels of CA19-9,CA242,CEA into the regression model,the OR value and 95%CI were 1. 717(0. 736 to 2. 359),2. 865(2. 217 to 3. 685),2. 614(2. 186 to 3. 127),3. 527(2. 842 to 4. 377),4. 214(3. 570 to 4. 962), 2. 315(2. 114 to 2. 539),1. 876(1. 175 to 2. 852). Conclusion: Serum tumor markers combined detection can help to improve the accuracy of early diagnosis of pancreatic cancer and smoking,incorrect diet habits,history of diabetes,biliary disease history and high levels of CA19-9,CA242 and CEA are independent risk factors for pancreatic cancer.

AIM:To compare the synthesis efficiency of n3 and n6 very long chain polyunsaturated fatty acid ( VLC-PUFA ) by overexpressing ELOVL4 protein, providing guidance for treating Stargardt-like macular dystrophy (STGD3).
METHODS:To establish recombinant adenovirus with the ELOVL4 protein and green fluorescent protein, transferred into cultured PC12 cells. The cells were divided into 3 groups: PC12, PC12 + Ad- GFP and PC12 + Ad-ELOVL4, former two groups serve as controls. ELOVL4 gene expression was quantified by qRT-PCRs. ELOVL4 protein was analyzed by Western - Blot ( WB ) . The transduced cells were treated with both EPA and AA (1:1). After 48h of incubation, cells were collected, total lipids extracted and fatty acid methyl esters prepared and analyzed by gas chromatography-mass spectrometry ( GC-MS) .
RESULTS:When supplemented together, 20:5n3 (EPA) and 20:4n6 ( AA) were efficiently taken up at almost the same amounts in the PC12 cells regardless of ELOVL4 expression. The ELOVL4-expressing cells elongated both EPA and AA to a series of n3 and n6 VLC-PUFAs. From 20:5n3/EPA, 34:5n3 and 36:5n3 account for 0. 71% and 1.6%, respectively. From 20:4n6/DHA, 34:4n6 and 36:4n6 were only 0. 46% and 0. 61%, respectively. The total relative mol% of n3 VLC-PUFAs synthesized from EPA was almost two times that of n6 VLC-PUFAs synthesized from AA.
CONCLUSION: ELOVL4 protein preferentially elongates n3 PUFA to VLC - PUFAs over n6 PUFA. Dietary supplementation of appropriate n3/n6 PUFAs may provide STGD3 patients with some therapeutic benefits.

Objective To assess the effects of hepatitis B virus(HBV) large envelope protein (LHB) on the apoptosis of HepG2 cells and explore the possible mechanism by proteomic approaches.Methods LHB gene was cloned into pShuttle-IRES-hrGFP-1,and the recombinant adenovirus either barboring LHB(Ad-LHB) or empty vector(Ad-GFP) were separately generated.Annexin V-FITC apoptosis detection kit,JC-1 mitochondrial membrane potential assay kit and propidium iodide(PI) staining kit were employed combined with flow cytometry to detect the apoptotic cells infected with Ad-LHB or control of Ad-GFP.The cellular proteins were collected after infection of HepG2 cells by Ad-LHBs or Ad-GFP,and a total of 600 μg proteins were submitted to two-dimensional gel electrophoresis(2-DE) and stained with R350.The gel images were captured by ImageScanner Ⅱ Imaging System,the differentially expressed proteins were identified by ImageMaster 2D Platinum analysis software and picked up by Ettan Spot Picker.After enzyme digestion,the protein samples were analyzed by MALDI-TOF-TOF MS.Results HepG2 cells infected with Ad-LHB were much more prone to apoptosis.There were thirty nine differentially expressed proteins were determined by 2-DE between HepG2 cells infected with Ad-LHB and Ad-GFP,and they were identified ultimately and categorized into thirty three kinds of proteins by MALDI-TOF-TOF MS.Among these proteins,nine were found to be closely related to cell apoptosis,in which CAPN2,eIF3K and PPP2CB were higher expressed in Ad-LHB infected HepG2 cells,and SERPINH1,LASP1,PRDX1,DHRS2,LDHA and PS-MA4 were lower expressed in Ad-LHB infected HepG2 cells.Conclusion LHB could induce apoptosis of HepG2 cells,and several apoptosis-related proteins participated in this process.

Objective To observe the effects of co-transfection of proto-oncogene c-myc antisense oligodeoxynucleotides(c-myc-AODN) and tissue-type plasminogen activator(tPA) gene on intimal hyperplasia of auto-transplantion artery in rabbit. Methods The left and right external iliac artery(length 1.0 cm) of rabbits were cross transplanted. The artery grafts and sutures were respectively soaked in Lipofection, c-myc-AODN, pBudCE4.1/tPA, c-myc-AODN and pBudCE4.1/tPA solution for 15 minutes. Each group were divided into five subgroups(n=5, in each subgroup) according to the sacrifice times(3, 7, 14, 28 and 56 d after operation). Specimens were harvested for pathology, chromogenic substrate test, 3H-TdR incorporation test and immunochemisty coloration study. Result The intimal area, stenosis ratio, 3H-TdR incorporation, PCNA positive cell in c-myc-AODN adding tPA co-transfection group were significantly lower than that of control group(P0.01), and that were lower than c-myc-AODN transfection group and tPA gene transfection group(P0.05). Conclusion Vascular local co-transfection of tPA gene and c-myc-AODN effectively inhibits the proliferation of vascular smooth muscle cell(VSMC) and hyperplasia of intima of the transplanted artery.

In order to estimate the relationship between iron and the Alzheimer's disease, the behavioral and pathological changes were observed by Morris water maze and immunohistochemical staining respectively after injecting FeC13 into brain ventricle of rats. The apoptosis was tested by flow cytometry and the electron microscopy was used to observe ultrastructural changes. There were significant differences in escape latency of time and distance between normal animals and iron treated rats. Percentage and fluorescence intension (FI) of AnnexinV FITC loaded cells undergoing apoptosis were higher in iron treated rats compared with normal animals. Fawn-coloured products of β amyloid protein were interspersedly distributed in extensive areas of cerebral cortex and hippocampus. Under electron microscope, vacuolate degeneration of neuronal processes with mitochondria degeneration and accumulation of microtubule near vacuolar nucleus were observed in iron treated rats. These results suggest that a local higher concentration of iron in brain may induce Alzheimer-like impairment of intelligence and pathological changes.

Objective：To study the mental health status of heroin addicts and their need for psychotherapy after detoxification. Methods：The mental health of 83 heroin addicts and 70 normal subjects were assessed by SCL-90, Social Support Rating Scale , EPQ, SAS and SDS. Results：As compared to normal controls, heroin additions were found to score higher on various mental health measures, including SCL-90 total and factor scores, the N score of EPQ, as well as total scores on SAS and SDS. The social support in heroin addicts scored lower than normal subjects. Conclusion：Heroin addicts have severe psychological problems, which warrant long-term psychological rehabilitation after detoxification.

Objective To observe the influence of hands-washing and non-hands-washing on clinical disinfection effect before surgical hands antisepsis.Methods Totally 40 surgcal staff wee divided into the handa-washing group and the non-hands-washing group.The sterilizing effect of handa at diggerent time points during operations and kinds of bacteria colony were compared between two groups.Results Bacteria colony of hands at different time points during operations showed no significant difference between two groups.Both groups were detected bacteris colony on their hands,mainly made up of coagulase negative staphylococcus,corynebacterium and eaterococcus and no difference was seen between two groups.Conchusions The non-hands-washing method can acquire the same disinfection effect as traditional hands-washing method.Its disinfection effect lasts three hours,easy to operate,little stimulation on the skin,and was worthy of clinical application.

Objective To establish the animal model of acute intracranial hypertension,investigate the changes of transeranial Doppler patterns in raised intraeranial pressure so that the necessary experimental evidence will be provided to clinical experience for monitoring intraeranial pressure (ICP) and cerebral perfusion pressure (CPP) using transeranial Doppler non-invasively. Methods Acute intraeranial hypertension was induced by inflating the balloon inserted into the epidural space. Blood flow velocity of basilar artery was measured with transcranial Doppler and intracranial pressure as well as cerebral perfusion pressure was measured. Results With progressive increase of intracranial pressure five characteristic flow patterns were observed, appearing in the following order: high resistance pattern, systolic flow, retrograde diastolic flow, very small systolic flow and zero flow. Conclusions Analysing patterns of TCD may be helpful for evaluating ICP and CPP qualitatively. The animal model is simple and strict so that it is worthy of being spreadcd.