Objective:To analyze the sympathy,responsibility and rational emotion of clinical trainees and to construct the medical ethics emotion education system.Methods:A questionnaire survey was conducted among undergraduate clinical medical students.Of the 307 selected students,302 returned valid questionnaire.After eight months in the field trip,the educational outcome was assessed using questionnaire survey again,and the data was analyzed with descriptive statistics analysis.Result:According to the results of questionnaire survey,we came up with effective countermeasures to improve the medical ethics emotion of clinical medical students,and then strengthened students' sympathy,responsibility and rational emotion quality education.Conclusions:It suggests that cultivating the students' learning interests and motivating the subjective initiative of trainees on ethics learning with various teaching methods promoted the transformation from medical ethics emotion to behaviors and customs,and thus constructed a medical ethics emotion education model and improved quality medical ethics education.

Objective:Toprepare poly-DL-lactide-poly (PELA) microspheres encapsulating recombinant tissue inhibitors of metalloproteinase-1 (TIMP-1) adenovirus, and to investigate their effects on the proliferation of hepatocellular carcinoma HepG2 cells. Methods: The microsphere was constructed by encapsulating recombinant adenovirus containing TIMP-1 in biodegradable PELA. The diameter of the microsphere, quantity of virus encapsulated, loading rate, and releasing kinetics were measured. HepG2 cells were infected with the microspheres; the infection efficiency was examined by fluorescent microscope; and the ultrastructure was observed by TEM. The expression of TIMP-1 mRNA in HepG2 cells was examined by semi-quantitative RT-PCR, and the proliferation of HepG2 cells was detected by MTT assay. Results: The microsphere encapsulating recombinant TIMP-1 adenovirus was successfully constructed, with its diameter, entrapment efficiency, and virus loading rate being 1.965, 60.0%, and 10.5×10~8/mg, respectively. About 60% of the viruses were released within 120 h, and the total releasing time was longer than 240 h. Infection with rAdTIMP-1 PELA microsphere efficiently induced TIMP-1 expression in HepG2 cells, and significantly inhibited the proliferation of HepG2 cells, with the inhibitory rate being 47%. Conclusion: PELA microsphere encapsulating recombinant TIMP-1 adenovirus can markedly inhibit the proliferation of HepG2 cells, which provides an experimental basis for the combining macromolecular chemistry and gene therapy for treatment of hepatocellular carcinoma.

Objective To prepare recombinant human prothrombin-2 expressed in Pichia pastoris, and assay the enzymatic and clotting activities of prothrombin-2 activated by prothrombin activator ecarin.Methods Human prothrombin-2 gene and Echis carinatus ecarin gene were synthesized separately on the basis of the cDNA sequences published in GenBank.The gene of prothrombin-2 was cloned into the expression vector pPICZαA.The expression vector pPICZαA/prothrombin-2 was transformed into glycoengineered P.pastoris, and then prothrombin-2 engineered P.pastoris was screened.The expression products were induced by methanol, purified by two-step chromatography and identified by diges-tion by PNGase F and analysis of pepetide fingerprint.The ecarin gene was cloned into the expression vector pcDNA3.1. The expression vector pcDNA3.1/Ecarin was transformed into HEK 293T cells and the culture supernatant of HEK 293T/Ecarin was collected.The reaction product of HEK 293T/Ecarin cell culture supernatant and purified prothrombin-2 was analyzed by S-2238,which was the chromogenic substrate for thrombin.Fibrinogen was used to measure blood clotting time. Results The purified protein of P.pastoris expressed prothrombin-2 culture supernatant was 37 ×103 .The relative molecular mass(Mr) of the purified protein was reduced to 35 ×103, which was consistent with the theoretical Mr of prothrombin-2 molecular weight.The purified protein was proved to be prothrombin-2 by peptide fingerprint identification. The purified prothrombin-2 processed by HEK 293T/Ecarin culture supernatant could hydrolyze S-2238 to produce yellow pNA, and D405 of pNA increased with the volume of the processed prothrombin-2 that could promote the plasma coagulation.The blood clotting time was close to that of the thrombin kit.Conclusion Prothrombin-2 is prepared by P.pastoris and activated toα-thrombin by ecarin.This technique may replace the method of extraction of prothrombin from plasma and can be used for the treatment of war wounds or for future clinical research.

Objective To evaluate recombinant adenovirus microspheres encapsulated with antisense MRP(as-mrp) on reversion of hepatocellular carcinoma.Methods The rats were divided randomly into 5 groups:control group;hepatic arterial injection of normal saline group,microspheres carrying no viruses group,rAdV carrying as-mrp group,and microspheres with encapsulation of as-mrp rAdV(MER) group.The theraputic effect were observed.Results The tumor growth inhibition and the mean life time in MER group were superior to than of other 4 groups(P

Objective To investigate the different effect of exogenous leptin on estrogen receptor α,β mRNA in human breast tumor tissue in nude mice xenograft models. Methods We made nude mice xenograft models of MCF-7 human breast cancer cells cultured in vitro, then divided them into experimental group of]eptin( n = 30)and control group of normal saline( n = 30)randomly. The models of experimental group were injected subcutaneously the recombinant human leptin for 15 consecutive days, the models of control group were injected subcutaneously the same dose of normal saline. A real- time quantitative RT- PCR assay was developed to quantify the expression of estrogen receptor α, β mRNA in tumor tissue, using the relative quantitative analysis. Results The leptin-intervened nude mice xenograft models were safely established. The relative quantitation of estrogen receptor α mRNA was significantly higher in the leptin group than in the normal saline group ( P ＜ 0.01 ), the relative quantitation of estrogen receptor β mRNA was significantly lower in the leptin group than in the normal saline group ( P ＜ 0. 01 ). Conclusion The nude mice xenograft models can be safely intervened with human leptin by subcutaneous injection around tumor.Estrogen receptor is one of the targets of leptin in the progress of breast cancer. Exogenous human leptin can up- regulate the expression of estrogen receptor α and down- regulate the expression of the estrogen receptor βin nude mice xenograft models of human breast tumor.

Objective: To investigate the effects of silk fibroin on the immobilization of thrombin. Methods: The immobilized thrombin was prepared using silk fibroin as the carrier and glutaraldehyde as the crosslinking agent. With activity yield as the index, the process conditions of silk fibroin immobilized thrombin were determined by an orthogonal test. Results:The optimum process con-ditions of immobilized thrombin treated with silk fibroin were as follows:the immobilization time was 6 h, the enzyme dosage was 2 400 NIH·g-1 casein, the temperature was 25℃ and pH was 7. 6. The activity recovery of immobilized thrombin was 67. 22%. Conclu-sion:Silk fibroin has the positive immobilization effect on thrombin.

DCD WeChat ethical review had its necessity and superiority but problems and confusion as well. For instance, WeChat review time was urgent and limited;WeChat review discussed insufficiently even hard to car-ry out;the methods of WeChat review were limited;the authenticity of audit opinion was difficult to grasp;the re-view data privacy existed hidden danger;the review quality was difficult to guarantee;WeChat conference content file archive was difficult, and so on. As to this, the author put forward the following countermeasures:giving war-ranty on the ethical review time as far as possible;initiating the committee review by the referee and uploading the review information to WeChat group, then ethical reviewers expressing their opinions in real names to ensure data confidentiality and privacy protection; strengthening the ethics committee education for organ transplantation; im-proving the quality of the review committee members and moral cultivation;archiving the WeChat conference infor-mation.

Objective:To detect and verifica the gene profile difference of microRNA-146a (miR-146a) and its role in the pro-liferation of vascular smooth muscle cells (VSMCs) by gene chip technology. Methods: Artificially synthesized miR-146a mimics(50 nmol/L) ,miR-146 inhibitor ( 50 nmol/L ) , scramble ( 50 nmol/L ) and PBS were transfected into cultured primary rat VSMCs in vitro. After transfection,Real time PCR was used to measure the levels of miR-146a and the cell counting kit 8(CCK8) was employed to investigate the proliferation of VSMCs. The VSMCs interfered by miR-146a inhibitor or miR-146a control were examined by gene chips and the profile of gene were analyzed by bioinformatics technology to detect the different genes and signal transduction pathway. The changes in mRNAs and proteins were accessed separately by Real time PCR and Western blot. Results: Compared with sham and control VSMCs,miR-146a expression level was significantly decreased in treatment with miR-146a inhibitor(P<0. 01),as well as optical density(OD) was also shown remarkably down regulated simultaneously(P<0. 05). The investigation of gene profile revealed that the p53 signal pathway was up-regulated in VSMCs interfered by miR-146a. The mRNA and protein expression levels of p53, caspase3 and PTEN in p53 signal transduction pathway didn′t show significant differences(P>0. 05),however,the mRNA and protein expression levels of cyclin D1 significantly increased in treatment with miR-146a mimics VSMCs group and decreased in miR-146a inhibitor VSMCs group ( compared with sham VSMCs group, both P<0. 05 ) . Conclusion: Our data indicated that miR-146a may promote the proliferation of rat VSMCs by up-regulating cyclin D1 expression.

Objective To retrospectively analyze the surgical methods and efficacy in 70 cases of type A aortic dissection patients over 65 years old.Methods From January 2005 to May 2012,70 type A aortic dissection patients over 65 years old received surgical treatment.Among them,there were 47 males aged 65 to 78 years old with mean 71,23 females,aged 65 to 72 years old with mean 68.55 cases were acute onset,while 15 cases were chronically onset.Different surgical methods were selected depend on patients' situations.We followed up all patients after discharged from hospital to continue to observe their health situation and evaluate the therapeutic effects.Results After surgery,eight patients died in the hospital,62 patients were recovered and discharged from the hospital.The mortality rate is 11.4％.During the follow up period from 3 to 72 months,there were no dead,aneurysm rupture and others severe complications.9 cases received endovascular graft exclusion within 6 months after discharged from hospital.The survival patients were satisfactory healed with their daily living activity resumed.Conclusion For over 65 years old patients with aortic dissection,the accurate and rapid selection of surgical method could improve the survival rate and the quality of life with a lower occurrence rate of complications.

Abstrac:Aim To study the effect of hydroxysafflor yellow A ( HYSA ) on the proliferation of vascular smooth muscle cells ( VSMCs) and the related molecu-lar mechanism. Methods The inhibitory effects of hydroxysafflor yellow A on VSMC proliferation was de-tected using cell culture, MTT assay, Western blot and immunohistochemical staining. Results The results showed that HYSA inhibited cell proliferation induced by PDGF in a dose-dependent (5,10,20,40 μmol· L-1 ) manner, reduced proliferating cell nuclear anti-gen ( PCNA ) expression and blocked PDGFR-MEK-ERK1/2 signaling pathway activated by PDGF in VSMCs. Conclusion HYSA inhibits VSMCs prolifer-ation via reducing the expression of PCNA and blocking signal transduction of MEK-ERK1/2 in VSMCs.