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 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.

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.

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.

Objective To understand the influencing factors of professional adaptability for college nursing students and the mechanism. Methods A group of 1 105 college students from grades 2010 and 2011 were surveyed with questionnaire for professional adaptability. The data were analyzed with SPSS 16.0. Measurement data were expressed as x±s while enumeration data were expressed as percentage. t test and variance analysis were used to compare the differces among groups of different grades and genders. Results The professional adaptability mean score of college nursing students was (102.66 ± 14.93) and significant differences in professional adaptability were observed between different grades and genders (P=0.004 3, P=0.001 5). The possible influencing factors professional adaptability were different sources of students, family location, family income, etc. There were statis-tic differences in professional promise between arts and science students and between rural an urban students (P=0.029 8, P=0.007 0). Conclusion Grade and genders are the main influencing factors for professional adaptability of college nursing students while other factors exert little effect on it.

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 investigate the influence of CD4+CD25+ regulatory T cells (Tregs) on the disease progression in MRL/lps mice. Methods Tregs were separated by using magnetic beads from splenic cells of MRL/lps mice and BALB/c mice, and concentrated. Twenty-four MRLAps mice were equally divided into 3 groups, test group 1 injected with Tregs from MRL/lps mice, test group 2 injected with Tregs from BALB/c mice, and control group injected with physiological sodium chloride solution. Three weeks later, the levels of urine protein as well as serum anti-dsDNA antibody were determined; subsequently, the mice were sacrificed followed by histopathological and immunopathological examination of renal tissue. Results A significant decline was observed in the test group 1 compared with the test group 2 and control group in the urine protein score (10.63 ± 4.17 vs. 20.00 ± 5.35 and 18.75 ± 8.34, both P < 0.05), serum anti-dsDNA antibody level (5.36 ± 2.40 pg/ml vs. 9.57 ± 1.97 pg/ml and 10.75 ± 3.98 pg/ml, both P < 0.05), glomerular sclerosis index [(32.00 ± 12.09)% vs. (45.50 ± 13.68)% and (47.50 ± 10.78)%, both P< 0.05], and immunofluorescence intensity of IgG immune complex in renal tissue (1.88 ± 0.99 vs. 2.88 ± 0.64 and 2.75 ± 0.71, both P< 0.05). No significant difference was noted in renal tubule interstitial impairment index between the 3 groups (4.63 ± 1.92, 6.00 ± 1.07 and 5.75 ± 1.28, all P> 0.05). There was no statistical difference between the test group 2 and control group in terms of any of the above parameters (all P > 0.05). Conclusions Injection of Tregs from homologous mice could significantly down-regulate proteinuria degree, serum anti-dsDNA antibody level, glomerular sclerosis index and IgG immune complex level in renal tissue, and thereby decelerate the progression of renal impairment in MRL/lps mice.

BACKGROUND:The important reason for failed back surgery syndrome is the postoperative epidural scar adhesions, therefore, exploring the methods of preventing postoperative lumbar epidural scar adhesions has always been a hot research in spine surgery field. OBJECTIVE:To investigate the effect of Shenshu point magnetic stimulation therapy on epidural scar adhesions in rat models of failed back surgery syndrome. METHODS:Sixty Sprague-Dawley rat models of failed back surgery syndrome were successfuly established using the method of laminectomy, and then divided into Shenshu magnetic therapy group and blank control group. Beginning from 1 week after modeling, rats in the Shenshu magnetic therapy group were subjected to bilateral Shenshu magnetic stimulation for 6 weeks, 5 days of treatment per week. Rats in the blank control group were not given any intervention. At 13 weeks after modeling, rats were harvested and the area ratio of epidural scars, range of adhesions and formation of colagen fibers were observed under light microscope. Fibroblasts were counted. Hydroxyproline content and transforming growth factor β1 expression in scar tissue were compared between these two groups.