BACKGROUND:Although the methods of mechanical scrapping,tissue block transplantation,and enzyme digestion to culture endothelial cells have been quite mature,it has been constantly updated.OBJECTIVE:To explore the culture and identification of rabbit aortic endothelial cells.METHODS:Aorta was derived from one-week-old New Zealand rabbit.Following removing the adventitia,the endothelium was digested in the mixture of 2 g/L type Ⅰ collagen,2 9/L type Ⅱ collagen,2 g/L type Ⅳ collagen,and 2 g/L type Ⅴ collagen (1:1:1:1)for 20 minutes,and the digestionwas stopped with culturemedium(1:1).Cells in the endothelium were lightly scraped to make cell suspension,which was centrifuged to precipitate cells with DMEM culture medium containing 20%fetal bovine serum,1 μg/L VEGF,2 μg/L bFGF,and 6 U/L gentamicin.The cells were then blown into single cells and the culture liquid was changed every 48 hours.The cells were passaged according to the ratio of 1:2.Inverted phase contrast microscope was used to observe cell culture;immunohistochemistry and immunofluorescence were used to determine Ⅷ-related antigen;electron microscope was used to detect Weibel-Paladed body.RESULTS AND CONCLUSION:Ⅷ-related antigen and electron microscopy confirmed that the primary and 5-passage endothelial cells were successfully cultured,suggesting that rabbit aortic endothelial cells were cultured into cell line.Ⅷ-related antigen and electron microscopy determined that a combination with W-P body was a good method to determine endothelial cells.

Objective To compare serum protein fingerprint among Uygur patients with thyroid cancer and benign thyroid nodules, and Han patients with thyroid cancer, and to screen ethnic-specific protein markers of thyroid cancer. Methods Using the technology of two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), the protein expression profiles of Uygur and Han patients with thyroid cancer and Uygur patients with thyroid nodules were established and compared by image analysis software between two groups. Liquid chromatography-mass spectrometry/mass spectrometry coupling techniques ( LC-MS/MS) were used to identify differential protein. The protein′s name, cellular localization and functional classification were searched in Swissport database. Results There are 11 differentially expressed proteins by comparison of sera in Uygur patients with benign thyroid nodules and thyroid cancer, in which complement C3 and C4b levels were down-regulated in the serums of Uygur patients with benign thyroid nodules, and 9 proteins such as heme-binding proteins etc were up-regulated. There are 7 differentially expressed proteins by comparison of serums in Uygur and Han patients with thyroid cancer, in which transferrin level was up-regulated in the serum of Uygur patients with thyroid cancer, and 6 proteins such as cytokeratin-1 etc were down-regulated in serum of Uygur patients with thyroid cancer. Conclusion Combined screening of multiple labelled proteins including heme-binding protein,α2-macroglobulin, and transferrin protein etc may provide the basis for the diagnosis of thyroid cancer in Uygurs and Hans.

ObjectivesTo explore the serum insulin-like growth factor-1 (IGF-1)levels in the patients with unresectable or metastatic gastrointestinal stromal tumors before and after imatinib treatment and to discuss its clinical significance.MethodsThe serum samples of27 patients with unresectable or metastatic gastrointestinal stromal tumors were collected before imatinib treatment(before treatment group) and three months after imatinib treatment(after treatment group).The other serum samples were collected from 20 healthy volunteers who were accepting health examination (control group). The serum IGF-1 levels of the samples were determined by enzyme-linked immunosorbent assay (ELISA).ResultsThe mean serum IGF-1 levels were significantly higher in before treatment group than in control controls[(463.61±120.98)ng/ml vs.(115.75±39.27) ng/ml, t=12.355,P=0.000)]. Three months after imatinib treatment,the mean serum IGF-1 levels were significantly lower in after treatment group than in before treatment group [(244.64±100.11)ng/ml vs.(463.61±120.98) ng/ml, t=7.582,P=0.000].ConclusionsSerum IGF-1 levels may help to judge therapeutic effect,progression,recurrence or metastasis of the gastrointestinal stromal tumor.

Objective To investigate the effects of imatinib mesylate on recurrent or metastatic gastrointestinal stromal tumor.Methods Thirty-five cases of patients with recurrent or metastatic gastrointestinal stromal tumors treated with Imatinib mesylate from January 2007 to February 2010 in our hospital were analyzed retrospectively.Results After treated with Imatinib mesylate for 3 months,complete remission rate was 2.9％( 1/35 ),partial remission rate was 45.7％ ( 16/35 ),stable disease rate was 51.4％ ( 18/35 ) and progressive rate was 0;clinical benefit rate was 100％.The 2-year overall survival rate was 97.1％.The 2-year progression free survival was 82.9％.Conclusion Imatinib mesylate is effective for recurrent or metastatic gastrointestinal stromal tumors and the treatment is safe and reliable.

Objective To explore changes of serum fibroblast growth factor 2 (FGF2) levels after treatment with imatinib mesylate in patients with unresectable or metastatic gastrointestinal sromal tumor.Methods Serum FGF2 levels (with ELISA) were determined in 27 patients with unresectable or metastatic gastrointestinal stromal tumor both before and after treatment with imatinib mesylate.Results Before treatment with imatinib mesylate,of 27 cases of unresectable or metastatic gastrointestinal stromal tumor,17 cases (63.0 ％) were found positive for serum FGF2,after treatment with imatinib mesylate,9 cases (33.3 ％) were found positive for serum FGF2 (P ＜ 0.05).Using imatinib mesylate for 3 months,complete remission rate was 0,partial remission rate was 51.9 ％ (14/27),stable disease rate was 48.1％ (13/27) and progressive rate was 0.Conclusion Patients with unresectable or metastatic gastrointestinal stromal tumor have elevated serum FGF2 levels.Imatinib mesylate treatment can reduce serum FGF2 levels in patients with unresectable or metastatic gastrointestinal stromal tumor.

Objective The aim of this study is to provide guidance on rehabilitation nursing via observing the effect of comprehensive relaxation training on perioperative psychological stress in patients with coronary intervention treatment. Methods In this study, eighty patients ready to receive coronary intervention were randomly divided into the experimental group and the control group (40 patients in each). The control group were treated with routine percutaneous coronary intervention care, and the experimental group received both comprehensive relaxation training and routine care. The anxiety state and subjective feelings on comprehensive relaxation training in both groups were dynamically investigated via State-Trait Anxiety Inventory questionnaire and subjective feeling questionnaire from pre-operation to 7 days after operation. Results Anxiety levels between two groups showed the similar trend, and there was statistically significance between two groupsin the state anxiety level. The subjective feeling in the experimental group improved. Conclusions The psychological stress in patients receiving coronary intervention surgery maintains at high level form pre-operation to 24h post operation, and comprehensive relaxation training can significantly reduce the level of state anxiety. These patients consider that this simple training shows obvious relaxing effect.

Drug-induced liver injury is commonly seen in ctinical practice and is usually caused by antipsychotic drags,antitubercnlar agents,paracetamol,and statins.Silybin,a substance with biological activity in silymarin extract,has antioxidant,anti-inflammatory,and anti-fibrosis effects.This article summarizes the research advances in silybin in the prevention and treatment of drug-induced liver injury.

Objective:To investigate the effects of pristimerin therapy on the functions of Th17 and Treg cells in patients with allergic asthma,and expore the function of immune factors in therapy on allergic asthma. Methods:Extracted mononuclear cells in pe-ripheral blood of patients with allergic asthma,and given pristimerin intervention in vitro. Then used fluorescence-activated cell sorting analysis to detect the contect of Th17 and Treg,enzyme-linked immunosorbent assay to detect cytokine production of IL-17 and TGF-β, and Real-time PCR to detect the contect of RORγT and Foxp3. Results: Compared with the control group, experimental group had reduced expression of CD4+IL-17+T cells ( Th17 cells) ,and increased expression of CD4+CD25+Foxp3+T cells ( Treg cells) ,which all had the statistical significance (P<0. 05);displayed decreased level of IL-17,and increased level of TGF-β,which all had the statistical significance (P<0. 05);had reduced expression of RORγT,and increased expression of Foxp3,which all had the statistical significance (P<0. 05). Conclusion: Pristimerin therapy can remit the unbalance of Th17/Treg in vitro,hinting this drug may regulate allergic asthma by influencing immunologic balance,which offers the theoretical support for pristimerin in clinical application.

Objective To investigate the correlation between methylation status of UNC 5C gene promoter with colorectal cancer UNC5C .Methods 54 cases of sporadic colorectal cancer and related normal mucosal tissue ,as well as 6 colon cancer cell lines and fiber cell lines(FCL) in the oncology department of the Kailuan General Hospital from February 2010 to March 2013 were selected as the research objects and performed the mRNA and methylation analysis for exploring the correlation between the netrin‐1 receptor UNC5C defects with colorectal cancer disease .Results mRNA of UNC5A and UNC5B was expressed in the detected colorectal cancer cell lines .Except FCL for UNC5C ,all of the cancer cell lines had no mRNA expression .In colorectal cancer tissue , UNC5C methylation was significantly higher than that in the normal mucosa ,the difference was statistically significant (P<0 .05) . The methylation levels of UNC5C gene in the stage Ⅰ to Ⅱ ,and the stage Ⅲ to Ⅳwere significantly higher than non‐methylation levels ,the differences were statistically significant (P<0 .05) .The correlation analysis by the Spearman method showed that the UNC5C defects was positively correlated with colorectal cancer disease (r= 0 .856 ,P< 0 .05) .Conclusion Netrin‐1 receptor UNC5C defect has certain correlation with colorectal cancer disease .

Objective To explore the preventive effect of Bailing capsules(BL) on myocardial fibrosis in the viral myocarditis(VMC) mice models and to clarify the possible mechanism.Methods The models of VMC were established by injecting Coxsackie virus B3 (CVB3) solution into 50 BALA/C mice.The 31 mice were divided into model group(n =11),large dose BL group(n =10)and small dose BL group (n =10).At the same time the control group (n =10) was established.The mice in model group and control group were fed with water 1 mL/d,the mice in large dose BL group were fed with BL at a dose of 7.5 g/(kg · d),and the mice in small dose BL group were fed with BL at a dose of 2.5 g/(kg · d).After 60 d,the levels of angiotensin Ⅱ (Ang Ⅱ) in blood serum of mice in various groups were detected by using ELISA.Myocardium tissue of mice was stained by Masson and collagen volume fraction(CVF)was accounted.At the same time,the content of periostin mRNA and transforming growth factor β1 (TGF-[β1) mRNA in myocardium tissue of mice were detected by adopting reverse transcription(RT)-PCR.Results Compared with the control group,the levels of serum Ang Ⅱ,CVF,TGF-β1 mRNA and periostin mRNA were increased (all P ＜ 0.01) in the model group.Compared with the model group,the levels of serum Ang Ⅱ,CVF and TGF-β1 mRNA were obviously decreased in large dose BL group and small dose BL group (all P ＜ 0.05).Compared with the model group,the level of periostin mRNA was obviously decreased in large dose BL group (P ＜ 0.05),but the level of periostin mRNA in small dose BL group was not decreased obviously (P ＞ 0.05).Conclusions BL can offer some protection to myocardial fibrosis in VMC mice.The possible mechanism may be performed through inhibiting the expression of serum Ang Ⅱ and TGF-β1 in myocardium tissue to reduce the expression of periostin.