Abstract:Acute pancreatitis(AP) is characterized by its sudden onset and rapid progression and is often complicated by liver injury. AP-induced liver injury may develop into hepatic failure and even result in death. Thus, it is of importance to protect liver function and block injury-related pathways. In the pathogenesis of liver injury in AP, inflammatory cytokines, nuclear factor-kappa B(NF-κB) and oxygen free radicals play important roles. The complexity of the mechanism underlying the development of liver injury exerts, to some extent, a contribution to the difficulties in the treatment of this disease. Currently, the drugs used to treat the disease include L-arginine (L-Arg), calcium ion antagonists, somatostatin and a variety of inflammatory mediator inhibitors. Additionally, some traditional Chinese medicines such as tripterygium, wilfordii, rhubarb and salvia miltiorrhizae may also have some effects. In this article, the pathogenesis of liver injury in AP and its therapy are reviewed.

Abstract:Objective: To construct recombinant adeno-associated virus co-expressing human vascular epithelial growth factor 165(hVEGF165) and bone morphogenetic protein 7(hBMP7), measure the virus titer and verify the recombination. Methods:The AAV helper-free system was used as basis to generate recombinant AAV. The IRES sequence of plasmid pIRES was cut down and subcloned into ITR/MCS containing vector pAAV-MCS to construct recombinant plasmid pAAV-MCSa-IRES-MCSb. The hVEGF165 and hBMP7 gene was amplified by PCR and inserted into upstream MCSa and downstream MCSb respectively. Then, recombinant plasmid pAAV-hVEGF165-IRES-hBMP7, pAAV-RC and pHelper were co-transfected into AAV-293 cells to complete rAAV-hVEGF165-IRES-hBMP7 packaging. The GFP labeled rAAV-IRES-GFP was simultaneously packaged by using the parallel plasmid pAAV-IRES-hrGFP. The efficiency of AAV packaging was monitored under fluorescent microscope and recombinant viral particles were harvested from infected AAV-293 cells. The virus titer was measured by infecting AAV-HT1080 cells, and the recombinant AAV-hVEGF165-IRES-hBMP7 was verified by PCR of the exogenous interest genes. Results:Recombinant pAAV-hVEGF165-IRES-hBMP7 was verified by double digestion. GFP expression in AAV-293 could be observed under fluorescent microscope 72 h after transfection and the system provided a high packing ratio of 95%. The recombinant adeno-associated virus has a high titer of 5.5×1011vp/ml, and AAV-HT 1080 was infected at a ratio of 90%. The recombinant virus was confirmed by PCR of exogenous hBMP7 and hVEGF165 gene. Conclusion:Recombinant rAAV-hVEGF165-IRES-hBMP7 was successfully constructed with a high virus titer, which may offer foundation for in vitro and in vivo experiments of hVEGF165 and hBMP7 co-expression and provide a new method for gene therapy of bone regeneration.

Abstract:Objective: To study the role of nuclear factor-kappa B(NF-κB) in cholesterol efflux from THP-1 derived-foam cells treated with AngiotensinⅡ(AngⅡ). Methods:Cultured THP-1 derived-foam cells were treated with AngⅡ or preincubated with tosyl-phenylalanine chloromethyl-ketone(TPCK) NF-κB inhibitor. The levels of activated NF-κB in the cells were examined by sandwich ELISA. Cellular cholesterol content was studied by electron microscopy scanning and zymochemistry via fluorospectrophotometer and cholesterol efflux was detected by scintillation counting technique. ABCA1 mRNA and protein were quantified by RT-PCR and Western blotting. Results:Addition of TPCK to the cells before AngⅡ stimulation attenuated the response of NF-κB p65 nuclear translocation induced by AngⅡ and showed no peak in foam cells group and caused a reduction in cholesterol content and an increase in cholesterol efflux by 24.1%(P < 0.05) and 41.1%(P < 0.05) respectively, when compared with AngⅡgroup. In accordance, the ABCA1 mRNA and protein were increased by 30% and 19%(P < 0.05) respectively, when compared with AngⅡ group. Conclusion:AngⅡ can down-regulate ABCA1 in THP-1 derived-foam cells via NF-κB, which leads to less cholesterol efflux and the increase of cholesterol content with the consequence of the promotion of atherosclerosis.

Abstract:Objective:To explore the effect of bile salt and bile acid on cultured eternalized human gastric mucosa epithelium GES-1 cells. Methods:Cultured eternalized human gastric mucosa epithelium GES-1 cells were treated with media containing 6 different kinds of bile salts and 3 different kinds of bile acids and their mixture with different concentrations: GCDC(glycochenodeoxycholate), GDC(glycodeoxycholate), GC(glycocholate), TCDC(taurochenodeoxycholate), TDC(taurodeoxycholate), TC (taurocholate), LCA(lithocholicacid), CA(cholic acid), DCA(deoxycholic acid)(50μmol/L,250μmol/L,500μmol/L,1000μmol/L), DY(mixture of bile salts) and DS(mixture of bile acids)(250μmol/L,500μmol/L,1000μmol/L,1500μmol/L, 2000μmol/L), in comparison with the control group(in normal media without bile salts and bile acids). Cell proliferation was assessed by MTT(3-[4,5-Dimethylthiaolyl]-2,5-diphenyl-tetrazolium bromide) assay for 72 hours with different concentrations and the apoptotic cells were assayed by flow cytometry (FCM) with Annex V-FITC conjugated with propidium iodide(PI) staining for 24 hours with different concentrations(1500,2000μmol/L). Results:There was no significant difference in morphology and cell proliferation in GC group after 24~72 h. Low concentration(50μmol/L) of GCDC, GDC, TCDC, TDC and TC accelerated gastric epithelial cell growth in a dosage-time dependent manner. At middle concentration(250-500μmol/L), it showed positive effect after 24~48 h, while negative effect after 72 h. At high concentration(1000μmol/L), it accelerated gastric epithelial cell growth after 24h and show consistent inhibition even leading to necrosis after 48~72 h. LCA and CA showed a positive effect on the concentration of 50μmol/L after 24~72 h, while 250-1000μmol/L showed a trend towards apoptosis after 24~72 h. At 50-500μmol/L, DCA showed proliferation after 24 h and apoptosis after 48~72 h, but showed necrosis after 24~72 h at 1000 μmol/L. DY and DS could facilitate normal gastric mucosa epithelial cell growth at low concentration (250-500μmol/L), however at 1000-2000μmol/L the trend shifted from apoptosis to necrosis. FCM with Annexin-V conjugated with PI staining revealed that GCDC, GDC, GC, TCDC, TDC, TC, LCA, CA, DCA, DY and DS induced apoptosis of human gastric mucosal epithelial cells. They were all significantly higher than that of the control(P < 0.05), but there was no significant difference in GC group (P > 0.05). The bile salts induced apoptosis in a time-dose-dependent manner. Conclusion:Our results suggested that bile acid and bile salt is the trigger of injury in human gastric mucosal epithelial cells.

Abstract:Objective: To evaluate the effectiveness of regenerous tissue and bone substitute in autogenous tooth transplantation in the larger recipient socket. Methods:In 3 Beagle dogs, 18 incisors were transplanted to the recipient sockets, 2 mm wider mesio-distally. The regenerous tissue group, the bone substitute group and the control group contained 7, 7 and 4 teeth respectively. No additional material was used in control group. Clinical and radiographic examinations were done every month and were sacrificed 3 months later. Subsequently, decalcified sections were prepared for routine histological evaluation. Ordinal scores for root surface resorption were analyzed using the Kruskal-Wallis test. Results:All donor teeth survived. A statistically significant difference was found among all three treatment groups(P = 0.0001). The proliferating tissue in space positively affected the periodontal healing without any resorption. Inflammatory resorption of the root surface and formation of new bone were observed in the bone substitute group. Surface resorptions of the roots were found in the control group. Conclusion:Proliferating tissues enhance the regeneration of periodontal tissues in larger recipient sockets and prevent root resorption. Sinbone HT is beneficial for the stabilization of the transplanted teeth in larger sockets.

Abstract:Objective:To evaluate Wells, Kahn, St.Andréand Constans scores for the prediction of deep venous thrombosis in Chinese patients. Methods:One hundred and seventy-two patients, prospectively, blinded referred for evaluation with four clinical-score systems for suspected deep venous thrombosis, were examined by ultrasonography. Sensitivity, specificity, positive predictive value, nega-tive predictive value and receiver operation curves were calculated for four clinical scores. The difference between areas of the ROC curve for each of the scores was compared with others and reference line. Results:Forty-six of 172 patients had deep venous throm-bosis proven by sonography. The sensitivity, specificity, positive predictive value and negative predictive value for Wells score was 91.3%, 57.1%, 43.8% and 94.7%, respectively, for Kahn score; 65.2%, 71.4%, 45.5% and 84.9%, respectively, for St.Andréscore; 63%, 38.9%, 27.4% and 74.2% respectively, for Constans score; 95.7%, 34.9%, 34.9% and 95.7% respectively. Area under ROV curve of Constans score was 0.814, which was similar to that of Wells score, then followed by Kahn score and that of St.Andréscore was no difference with the reference line. Conclusion:Based on the results of our study, the sensitivity, negative prediction value and area under ROC curve are larger for Constans score and Wells score in Chinese hospitalized patients than that of Kahn score or St.Andréscore. Considering the aim of the clinical assessment, Constans score and Wells score are more efficient for Chinese hospitalized patients.

Abstract:Objective:To analyze the value of CT pulmonary angiography(CTPA) in assessing right ventricular dysfunction(RVD) after acute pulmonary embolism. Methods:Thirty-six patients with CTPA-confirmed PE who underwent ultrasonic cardiography(UCG) within the ensuing 24 hours were retrospectively reviewed. According to the severity of the disease, the patients were divided into the massive PE group(24 cases) and non-massive PE group(12 cases) respectively. CT scans were analyzed for findings suggestive of RVD. Scans were considered positive for RVD if the right ventricle was dilated(RVd/LVd＞1) or if the interventricular septum was straightened or deviated towards the left ventricle. Results were then compared with the results of UCG to estimate the value of CTPA in detecting RVD associated with PE. Results:In all cases, compared with UCG, the diagnostic sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, positive predictive value, and negative predictive value of CTPA was 84.61%, 78.26%, 3.892, 0.197, 68.75% and 90% respectively. Kappa value was 0.60, which suggested moderate agreement between CTPA and UCG in the whole level. In the massive PE group, the diagnostic sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, positive predictive value, negative predictive value of CTPA was 84.61%, 72.73%, 3.103, 0.212, 78.57% and 80% respectively. Kappa value was 0.58, which suggested moderate agreement between CTPA and UCG in the massive PE group. In the non-massive PE group, the diagnostic specificity of CTPA was 83.33%. By statistics, the value of RVd/LVd had significant difference between the massive PE and the non-massive PE group. Conclusion:CTPA can reliably detect RVD through the evaluation of cardiac morphology. However, this result requires confirmation using a larger prospective cohort study.

Abstract:Objective: To compare clinical outcomes and safety of eptifibatide or tirofiban in patients with acute coronary syndrome(ACS) undergoing percutaneous coronary intervention(PCI). Methods:Thirty-six patients with ACS(unstable angina/non-ST-segment elevation myocardial infarction, UA/NSTEMI) who underwent PCI were randomly divided into two groups to receive eptifibatide or tirofiban treatment. Eptifibatide or tirofiban was predominantly initiated in the catheter laboratory before the intervention. In-hospital and 30-day MACE outcomes; bleeding as well as platelet counting were investigated in those two groups. Results:No in-hospital and 30-day MACE event occurred in the two groups. The number of ischemia leads after treatment reduced compared to that before PCI in the two groups. There was improvement in the number of ischemia leads for 24 h after administration in the tirofiban group than those in eptifibatide group(4.21±2.46 vs. 3.89±3.31, P =0.03). The two groups showed no incidence of massive bleeding. Minor bleeding rates were 16.7% and 22.2% in the two groups respectively. Conclusion:Eptifibatide as an adjunct to PCI may further decrease the incidence of ischemia event in patients with ACS and improve the safety, but its long-term efficacy and side effects need further observation.

Abstract:Objective: It was known that neutrophils play an important role in ischemic-reperfusion injury. In this study we tested the effect and its mechanism of dipyridamole on neutrophils. Methods:Hydrogen peroxide(H2O2) production by neutrophils was determined using luminol amplified chemiluminescence and the percentage of activity was calculated by observing the uninhibited peak height. Results: Dipyridamole per se produced a concentration-dependent inhibition of H2O2 by formyl-MetleuPhe(fMLP)-stimulated neutrophils. Dipy-ridamole at a low concentration(0.3μmol·L-1) that per se affected neutrophils only slightly, enhanced markedly the effects of adenosine on neutrophils. On the other hand, dipyridamole did not alter the inhibitory effect of NECA(5’-N-ethylcarboxamidoadenosine) on neutrophils. However, propentofylline, a known inhibitor of adenosine uptake, also gotten the same result. Conclusion:Dipy-ridamole inhibited the production of H2O2 by fMLP-stimulated neutrophils. Dipyridamole at a low concentration enhanced the inhibi-tory effect of adenosine on neutrophils. The mechanism involved is probably due to the effect of dipyridamole on adenosine uptake.

Abstract:Objective: To investigate the regulation of High sensitive C-reactive protein(Hs-CRP) and WBC count in patients with coronary heart disease(CHD) by percutaneous transluminal intervention(PCI) and to discuss the mechanism of inflammatory reaction after coronary stenting. Methods:127 patients who received successful percutaneous transluminal coronary stenting, were divided into groups of stable angina(SAP), unstable angina(UAP), and acute myocardial infarction(AMI) according to their clinical types. Another 41 stable angina patients with more than 7o% of coronary artery tenosis who did not receive PCI served as control. Serum Hs-CRP levels and WBC count were determined before intervention, 3 days and 7 days post PCI and the data were analyzed statistically by t-test. Results: There showed no difference in clinical baseline characteristics between groups. The serum Hs-CRP level and WBC count was gradually raised in the UAP and AMI group(how about SAP group, and had no difference in CAG group and SAP group). After PCI serum Hs-CRP levels and WBC counts were significantly higher in the SAP group than in the coronary angiography group(CAG) at 3 days and had no difference at 7 days. In the UAP and AMI group, the serum Hs-CRP level at 3 days and 7 days declined obviously, however serum WBC count did not decrease apparently. Conclusion: The serum Hs-CRP level and WBC count elevate transiently after PCI. There are different inflammatory reactions in different types of coronary heart diseases after coronary stenting procedure.

Abstract:Objective:To research on consistency of identifying solitary pulmonary masses with CT. Methods:Three observers with different working backgrounds in imaging diagnosis individually interpreted the same group images of solitary pulmonary mass, by 12 indexes of objective signs. The differences in interpretation resulted in ante- and post-interpretations were assessed by the x2 test. The agreement of two interpretations from the same observer was confirmed with the kappa test. A double-blind method was adopted for analysis. Results:The agreement rates of ante- and post-interpreting from the three observers were respectively 82.65%(486/588) 80.27%(472/588) and 84.86% (499/588) while their interpreting results were generally accordant without significant difference(x2 = 4.975, df = 2, P = 0.083) however there was difference between the observer 2 and observer 3(x2 = 4.875, df = 1, P = 0.027). There were five indexes with k > 0.40 of ante- and post-interpreting results of the three observers, including clarity of nodule borderline, presence of sentus, uniformity of density, existence of cavity and calcification in pathological region, among them, the agreement rate of interpreting borderline and cavity was higher(k > 0.07); the blood vessel convergence poorer(0 < k ≤ 0.40); the other six CT signs of interpretation were slightly different. Conclusion:The ability to identify solitary pulmonary mass was inconsistent, and needs to be improved further.

Abstract:Objective:To investigate the expression of Survivin p53 and its relationship with apoptosis、proliferation in hepatocellular carcinoma(HCC). Methods:The expression of Survivin, p53 and the proliferation of tumor cells marked by proliferation cell nuclear antigen (PCNA) in 42 cases of HCC were assessed by immunohistochemical method. TUNEL was used to detect apoptosis. Results:Survivin protein was expressed in 30 of 42 cases of HCC(71.4%) and in 4 of 34 cases of adjacent cirrhosis tissues(11.8%). Expression of Survivin protein was negative in 10 cases of normal tissues. Survivin protein positive expression rate in HCC was significantly higher than adjacent cirrhosis tissues and normal tissues(P < 0.001). The increased Survivin protein expression in cancer was significantly associated with histological grade(P = 0.003), p53 protein(P = 0.013), survival time(P < 0.05) and the ratio of proliferative index to apoptotic index(P < 0.01), but was not significantly correlated with age, sex, clinical stage, tumor size, metastasis, AFP, HBs-Ag(P > 0.05).Conclusion:There is a marked increased expression of Survivin in HCC, which may play an important role in breaking the balance of proliferation and apoptosis of HCC cells. The correlation between Survivin and p53 expression in HCC indicates that cooperation between Survivin and p53 plays a certain role in occurrence and /or development of HCC.

Abstract:Objective:To observe the clinical effect of anterior decompression and reconstruction using titanium mesh with locking plates in the treatment of cervical spondylotic myelopathy. Methods:One hundred and twenty patients with cervical spondylotic myelopathy were treated by anterior decompression and reconstruction using titanium mesh with locking plates. There were 66 men and 54 women ranges in age from 37 to 72 Years(mean age, 58.3 years). The mean Japanese orthopedic surgery association(JOA) scale was 9.6 points before operation. Patients were followed up clinically and radiographically. Results:Having stood surgery well, the operation time ranged between 60~100 min and bleeding during operation ranged between 20~200 ml. There were no case of postoperative infection, recurrent laryngeal nerve palsy, or esophageal or tracheal laceration or rupture. The average follow-up period was 14.3 months(range, 12 to 24 months) in 96 who were followed up. At the last follow-up visit the mean JOA scale had improved to 14.4 points, reflecting an improve-ment of 4.8 points. The results were considered to be excellent in 87 patients, good in 25, fair in 6, and poor in 2. No hardware-related complications or adjacent segment degenerative changes were encountered during the follow-up periods. Stable bone union was observed in all cases and the average time required for fusion was 5.7 months. Conclusion:Titanium mesh filled with autologous bone graft can avoid the complications associated with harvesting bone from the iliac crest donor site. When combined with cervical anterior locking plate, it can obtain satisfatory clinical results for the treatment of cervical spondylotic myelopathy.

Abstract:There were a lot of bright spots in the report “the advice regarding to deepening reform of the medical and health”, which was announced in mid-October, in order to ask for the precious suggestions from the public. Among the report, the most prominent idea was to emphasize that the new round of reform in China should stick to government-led medical reform which not only decided by the nature of government and the task of the reform, but also directly responsed to the objective realities in the field of medical health such as the obvious disparities of public welfare, economy and society. The author suggested that government-led idea should promote the government to change the functions during the process of building a service-oriented government, fundamentally solve the problem which medical treatment was difficult and expensive.

Abstract:The administration for food safety was an important measurement of the insurance of safe food. But the administration degree of food safety nowadays was not so strong, therefore the article suggested that the administration of food safety should be strengthened in the management of the original food material, the quick response for the food safety emergency, the further enforcement of the law, the research for the preventions with adequate money, more rewards for the report on food safety, more international exchanges in food safety, the improvement of the administration pattern, more controls of the environmental pollutions and the guidance for food production and process with adequate policy.

Abstract:Chinese medicine is the Chinese national outstanding cultural heritage. It has been the indispensable influential role in the national medical care. However, there are still shortcomings and deficiencies at present about the legal protection of Chinese medicine. By introducing the legal system of Chinese medicine about the ancient China, the Republic of China as well as the current period and analyzing the problems and the legislative difficulties in the current legal system of Chinese medicine, the article tried to provide some related suggestions for promulgating the fundamental law of Chinese medicine.

Abstract:This article discussed extensively how to apply product responsibility in the infringement of blood product from the perspectives of law application, the principle of clarifying responsibility, the distribution of proof burden and the relief of responsibility. The author argued that the burden of proof of causality in this kind of infringement should be inverted because it was also belong to the medical infringement. The author also suggested that the burden of proof of the product defect should also be inverted due to the principle in distributing the proof burden and the Article 7 of Proof Regulation.

Abstract:The article defined the legal nature of consultation that the consultation was a kind of civil contract act with the characteristics of innominate contract, consensual contract, formal contract, and onerous contract, etc. The authers demonstrated the civil liability related to the doctor in the process of consulation, including the principles of imputation of contract liability and tort liability, the type of contract liability and tort liability, and the civil liability of the consultants.

Abstract:Construction of a reasonable system of urban land rights would help to make full use of urban land and prevent waste of land resources. The urban land rights of civil law countries and regions were usufruct on the ground, easement, easement for special people, right of pawn and so on. China should learn from the experience of other countries and combine with specific national conditions to construct own city land rights system.

Abstract:Although the new Property Law had defined land contract operation right as usufructuary right, related issues were far from resolved which caused a lot of contradictions and conflicts. Legislation should provide appropriate solutions for the healthy development of land contract rights such as further improvement of collective land ownership, clear function of the land contract rights, and smooth channels of relief. In addition, this kind of right had insurmountable disadvantages, so the law should provide some other choices for farmers to choose according to the actual situation.

Abstract:The article researched the factors which influenced patients’ satisfaction in questionnaires. It found that the most important factor was the result of the treatment. Doctors’ service also had positive influence on patients’ satisfaction. However, how much they paid for the treatment had no notable influence. Patients would like to pay more money for better treatment and would not change their demand for medical care by the price. Hospitals had no need to reduce their price for more patients. Doctors’ service had good effect on treatment. So, if the hospital wanted to improve patients’ satisfaction, they should at first bring up doctors’ ability on treatment, then improve their service. The medical treatment price could be more appropriate just by government’s surveillance.

Abstract:Objective: To study the domain influencing factors and their internal relations which exist in hospitalization expenses of the main systematic diseases in rural health institutions in Nantong, to explore the main causes of unreasonable expenses growth, and put forward some suggestions. Methods: Factor analysis method was used to analyze the various indicators of hospitalization expenses among the illness case first pages of rural second-level hospitals and first-level health services centers in Haian and Haimen in April 2005. Results: Factors of examination fee and laboratory fee, treatment fee and nursing fee, bed fee and check fee, other fee, were the four public factors influencing hospitalization expenses. Conclusion: Make clear correlation and characteristics of public factors, take comprehensive measures to control unreasonable rise in hospitalization expenses in rural area.

Abstract:To evaluate the immediate effectiveness of peer education on AIDS prevention and provide bases for AIDS prevention education. Methods: A survey was conducted in October 2007 among 4 500 students from 15 colleges and universities in Nanjing on students’ knowledge, attitude and behavior concerning AIDS. A training program for AIDS prevention peer education was carried out among 1 400 college students of Grade 1 to Grade 3 selected by the methods of stratified cluster sampling in these colleges and universities. A parallel comparison group was then made about the effectiveness of the peer education, students receiving training as the intervention group and another 2 000 students not receiving training as the control group. Results: Comparing the proportions of knowing AIDS’s relative knowledge, having positive attitude to safe sex behavior and AIDS patients, the intervention group was significantly higher than pre-intervention group and parallel control group(P<0.05). The comparison between pre-intervention group and parallel control groups showed no statistical significances on several survey items(P>0.05). Conclusion: Peer education is suitable for university students and has good immediate effectiveness. The application and generalization of this effective means plays a highly important role in preventing and controlling the spread of AIDS.

Abstract:The article analyzed the status quo of the new rural cooperative medical system (NRCMS) in Nanjing city. The results of the investigation showed that NRCMS covered the rural population that increased continuously and the financing level rose year by year. It had realized severe illnesses orchestration that was more than other illnesses as principle of the policy, but the funds surplus rate was higher. The hospitalization person distribution was unbalance in the different rank hospitals. The author suggested that the service behavior of the contract medical treatment organization should be normalized, medical practice in hospital should be pursued class by class, township health center should strengthen building up, and reimbursement method and the way of expenses to pay should be actively carried out. NRCMS management information system should be built up and improved.

Abstract:Academic power and administrative power are the two basic forms in the academic management of colleges and universities. How to coordinate the operation of the two powers and how to guarantee exercising academic power have become hot issues in academic circles. This article argued that the breakthrough for many problems lied in ensuring that the administrative power would run by law. Based on analyzing the status and functions of administrative powers and the existing problems in the operation of administrative powers, the author put forward some solutions to regulate the operation of administrative powers.

Abstract:This article analyzed the present situation of university administrators in the higher education activity and discussed the necessary of the administrators integrating into the higher education activity. On the basis of these, the authors pondered how to make the university administrators integrate into the higher education activity on multi-positions by the superior resources of higher education, and pursue both the higher education and the university administrators themselves to get better and more harmonious development.

Abstract:In this article Pearson correlation coefficient was used to correlation analysis and significant test for readership of university library. Based on the results, the author suggested that the university library would arrange a reasonable stock structure, strengthen building human resources, and carry out a wide range of information services.

Abstract:General practitioners have been born under the development of modern medical mode and primarily engaged in grass-roots community health care services. Community health service is developing rapidly in China, but the training mode of general practitioner is still inadequate. This article analyzed and compared general medical education modes among the other countries, so as to discover some problems and give some advices to general medical education in China.

Abstract:On basis of principle of teaching students according to their aptitude,the students were classed into group A,B and C and trained by bi-level teaching. The effection of theaching implied the bi-level teaching was favourable to activate the initiative and enthusiasm of study,and was of great advantage to the ability of clinic practices,so did the clinic thinking training. It was reasonable to popularize and persist in bi-level teaching in clinic practice in prosthetic dentistry.

Abstract:Objective:To construct PI3-K/Akt shRNA expression vectors. Methods:Eight 19~21 bp reverse repeated motifs targeting of PI3-K/Akt gene were synthesized and cloned into eukaryotic expression plasmid pGenesil-1. After being screened and sequencing confirmed,the recombinant plasmids were transfected into rat glomerular mesangial cells(GMCs),then the levels of P-Akt and T-Akt protein in rat GMCs were measured using Western blot to select the optimal shRNA. Results:It was verified by partial nucleotide sequencing and restriction endonuclease digestion that the constructed eukaryotic vector expressing shRNA of PI3-K/Akt were correct. Western blotting results showed that the optimal shRNA which could effectively silence the target genes were shPik3c3-2 and shAkt1-4 respectively. Conclusion:The eukaryotic vectors expressing shRNA of PI3-K/Akt were constructed successfully.

Abstract:Objective:To construct recombinant plasmids containing 3 T cell epitopes from ESAT-6 antigen of Mycobacterium tuberculosis and extra-cellular fragment of Flt3 ligand(FL) genes,and to express them in rat glomerular mesangial cells(GMC). Methods:The amino acid sequence of ESAT-6 antigen was analysed using predictive algorithms and 3 T cell epitopes were predicted. The oligonucleotide encoding and the linker Ala-Ala-Tyr(AAY) were synthesized and inserted into the bicistronic vector pIRES and pIRES-FL. The recombinant plasmids were transfected into GMC cells,the recombinant proteins expressing in GMC cells was examined by Western blot. Results:The recombinant plasmids were verified by sequencing and the recombinant fusion proteins were identified by Western blot. Conclusion:The multiepitope DNA vaccine from ESAT-6 antigen of Mycobacterium tuberculosis was constructed and expressed successfully.

Abstract:Objective:Preparation and characterization of monoclonal antibodies against human herpesvirus 6(HHV-6). Methods:Purified HHV-6 was immunized to BALB/c mouse,and the spleen cells of the immunized mouse were fused with SP2/0 myeloma cells. The positive clones which produced McAbs against HHV-6 were screened by ELISA,indirect immunofluorescence assay. And samples of saliva from 25 oral squamous cell carcinoma patients were tested for human herpesvirus 6 by nested PCR assay and ELISA. Results:Clones named as JA9 could secrete specific McAbs against HHV-6 were detected. Isotrip kit of mouse Ig McAbs showed that McAb was IgG1. Culture supernatant and ascitic titres of the antibodies were 1 ∶ 500,1 ∶ 8.0×104,when tested by ELISA respectively. Western-blot showed JA9 was bound to 75 ku protein of HHV-6. HHV-6 positive rates were 60% by nested PCR assay and 40% by ELISA in the salivas of oral squamous cell carcinoma patients. Conclusion:The McAbs against HHV-6 might be a useful tool for fundamental studing of HHV-6 and clinic laboratory detection.

Abstract:Objective:To express structural GST-LMP-1 fusion protein in E.coli,detecting and analyzing its antibodies in NPC patients’ sera of EBV associated malignancies with Western-blot. Methods:The plasmid pMD18-LMP1 containing the recombinant extracellular peptide gene of LMP1 of EBV was digested with BamHⅠ and EcoRⅠ,and cloned into expression vector pGEX-4T-2. The constructed vector which had been identified by enzyme digestion and nucleotide sequence analysis was transformed into BL21(DE3). After induced with IPTG,the recombinant protein was purified with GST affinity chromatography,and confirmed by Western blot. Results:The recombinant plasmid was constructed correctly. SDS-PAGE analysis showed that the recombinant protein was about 32.2 ku. The purified protein could be used as an antigen to detect specific antibodies in the sera of patients with NPC by Western blot. Conclusion:The recombinant extracellular peptide gene of LMP1 of EBV could be expressed with high performance,and the antigenicity of the purified protein was certificated in sera of patients with NPC, which was associated with EBV.

Abstract:Objective:To investigate the effect of epidermal growth factor receptor(EGFR) inhibitor AG1478 on adhesion,migration and invasion of HuH7 cell line and its molecular mechanism. Methods:HuH7 cells were treated with AG1478. Cell adhesion assay and Matrix-gel were used to examine the capability of adhesion,migration and invasion. Western blotting was employed to detect the expression of Erk2 and Paxillin in HuH7 cells. Results:AG1478(20 μmol/L) significantly inhibited the adhesion of HuH7 cells by 52%. And Matrix-gel experiment showed that AG1478(20 μmol/L) decreased the migration and invasion ability of HuH7 cells in vitro by 49% and 70%,respectively(P < 0.01). After the treatment of AG1478 for 24 h,the expression of Erk2 and Paxillin were decreased remarkably(P < 0.01). Conclusion:Our findings suggested that EGFR signal transduction pathway might play a key role on the capability of adhesion,migration and invasion in human HCC cell line HuH7,which could be partly related to the mechanism of Erk2 and Paxillin signal pathway.

Abstract:Objective:To establish Livin specific gene-silencing system and to obtain stable clones of SGC-7901/siRNA-Livin. Methods:The small interfering RNA eukaryotic expression vector specific to Livin was constructed by gene recombination,and the nucleic acid was sequenced. Then it was transfected into SGC-7901 cells by Lipofectamin 2000. RT-PCR and Western blot were used to validate gene silencing efficiency of Livin in SGC-7901 cells. Stable clones were obtained by G418 screening. Results:Restriction endonuclease analyses showed that all plasmids had been successfully recombined. Nucleic acid sequencing showed that the siRNA was synthesized and inserted into pGPU6/GFP/Neo exactly. Livin silenced SGC-7901 cell clones were obtained after gene transfection and G418 selection,while the silencing efficiency of pGPU6/GFP/Neo/Livin-4 was higher than 70%. Conclusion:Livin specific siRNA was successfully constructed,and the expressions of Livin was silenced. Stable clones were obtained after gene transfection and screening.

Abstract:Objective:To isolate and clone the Negative factor(Nef) gene of human immunodeficiency virus 1(HIV-1),and to investigate its expression in the BCBL-1 cells and NIH/3T3 cells. Methods:A pair of PCR primers for Nef gene with EcoRⅠ and SalⅠ restriction enzyme cut sites was designed according to the sequence registered in GenBank. Then the genome of plasmid PCINL was taken as template and Nef gene was amplified using PCR. Subsequently,amplified gene fragments were digested with the two enzymes mentioned above,and then cloned into pCI-neo vector to create recombinant eukaryotic expression plasmid designated as pNefF,which was introduced with Flag sequence to facilitate the detection of protein. After identification with enzyme digestion and nucleotide sequences analysis,recombinant pNefF DNA was transient transfected into BCBL-1 cells and NIH/3T3 cells. The expression of pNefF mRNA and protein in BCBL-1 cells was detected by RT-PCR and Western blot,respectively. Results:Nucleotide sequences analysis indicated that the isolated and cloned pNefF sequence length was 651 bp. The isolated K9 sequence was 100% homology with Nef gene previously registered in GenBank. The specific bands were both detected at the expected place by RT-PCR and Western blot. Conclusion:Nef gene could be correctly expressed in BCBL-1 cells and NHT/3T3 cells.

Abstract:Objective:To evaluate the expression level and the diagnostic role of GDF 15 as a potential tumor marker in small hepatocellular carcinoma(SHCC) in human beings. Methods:Based on the previous data of Affymatrix microarrays,which indicated an over-expression of GDF15 in early-stage HCC of rat models,transcription level of GDF15 was examined in both rat models and human SHCC by RT-PCR. Real-time quantitative PCR was used to quantitatively analyze the gene in ten human small hepatocellular carcinoma and ten normal liver tissues,respectively. The protein levels and expression sits were subsequently confirmed in the SHCC,single large hepatocellular carcinoma(SLHCC), paraneoplastic tissues and normal liver tissues of human by immunohistochemistry(IHC),respectively. Results:GDF15 expression was significantly higher in the rat and human SHCC than in the normal liver tissues(P < 0.05). Real-time quantitative PCR and IHC analysis revealed that GDF15 expression was also higher in SHCC than in the paraneoplastic liver tissues and normal liver tissues. GDF15 protein detected in all 12 SHCC was mainly observed in the cytoplasm and extra cellular matrix of tumor cells. Conclusion:The current results showed GDF15 expression was high in the SHCCs and suggested it should be a useful tumor marker in detecting SHCC.

Abstract:Objective:To detect the expression and distribution of aquaporin1(AQP1) in adenocarcinoma of the lung and to analyze its clinical significance. Methods:The lung tissue of 30 cases of pulmonary adenocarcinoma and 30 normal cases were sampled immediately after surgery to study the expression and distribution of aquaporin1 in adenocarcinoma of the lung by immunohistochemical and Western blot analysis. Results:The sporadic expression of AQP1 was found in the vascular endothelial cells of normal lung tissues. The majority of AQP1 expression could be detected in the vascular endothelial cells,tumor cells in adenocarcinoma of the lung;the expression of AQP1 protein in tumor tissues was remarkably higher than that in normal tissues beside tumors;the expression of AQP1 protein in tumor tissues was correlated with tumor grading and lymph node metastasis. Conclusion:The expression of AQP1 was increased in adenocarcinoma of the lung over normal lung tissues. It suggested that AQP1 might play an important role in the formation and development of adenocarcinoma of lung and the mechanism need to be further studied.

Abstract:Objective:To investigate the expression of EGFR and COX-2 and their correlations in human gastric carcinoma,and analyze their relationship with H.pylori infection. Methods:By Immunochemical EnVision method,the expression of EGFR and COX-2 were detected in 47 cases of gastric cancer and 20 cases of cancer-adjacent tissues. The statuses of H.pylori infection were detected by fast urea enzyme test and 13C breath test. Results:The positive rates of EGFR and COX-2 in gastric carcinoma were significantly higher than those in paraneoplastic tissues(P < 0.01). There were significantly positive correlation between the expression of EGFR,COX-2(P < 0.05),and the expression with H.pylori infection were higher than those without H.pylori infection(P < 0.01). Conclusion:EGFR and COX-2 were overexpressed in gastric carcinoma,and there were positive correlations between them. H.pylori infection could upregulate the expression of EGFR and COX-2 in gastric cancer tissues.

Abstract:Objective:To investigate the association of ERCC1 mRNA levels and ERCC1 118 C/T polymorphism with the clinical outcome of gastric cancer patients treated with oxalipatinum-based adjuvant chemotherapy. Methods:Sixty-two gastric cancer patients received oxalipatinum-based adjuvant chemotherapy. The intratumoral ERCC1 mRNA expression was tested by semi-quantitative RT-PCR, and ERCC1 118 C/T polymorphism was tested by PCR-LDR(ligation detection reaction) method. Results:The median ERCC1 RNA level in 62 gastric cancer tissues was 0.672. The frequencies of ERCC1 118 C/C,C/T and T/T genotype were 51.6%(32/66),43.5%(27/62) and 4.8%(3/62),respectively. The recurrence-free and overall survival rate in patients with low level of ERCC1 mRNA(>0.672)were significantly higher than those in patients with high level(P < 0.05),while there was no significant difference found between patients with ERCC1 118 C/C and variant genotypes(T/T or C/T). Multivariate analysis showed that ERCC1 mRNA level was a potential predictor for relapse and survival in gastric cancer patients treated with platinum-based adjuvant chemotherapy(P < 0.05). ERCC1 118 C/T polymorphism was not associated with the disease prognosis. Conclusion:The ERCC1 mRNA level might be a useful predictive parameter for the relapse and survival of gastric cancer patients received with oxalipatinum-based adjuvant chemotherapy.

Abstract:Objective:To investigate the relationship between polymorphisms of DNA repair gene XPD751 and sensitivity to Gemcitabine/Cisplatin(GP) chemotherapy in advanced non-small-cell lung cancer(NSCLC). Methods:116 patients with NSCLC were analyzed. All patients were treated with GP chemotherapy,and DNA of peripheral blood leukocytes was obtained before therapy. XPD751 genotypes were detected by PCR-RFLP method. Results:①Of all cases,the frequencies of XPD751 Lys/Lys,Lys/Gln and Gln/Gln genotype were 81.0%,17.2% and 1.8%,respectively. The overall response rate of chemotherapy in all patients was 37.9%. ② The response rate of chemotherapy among patients with XPD751 Lys/Lys,Lys/Gln and Gln/Gln genotypes were 39.4%,25.0%,100.0%,respectively,and no significant difference was found(χ2 = 4.775,P = 0.093). Comparing to patients with Lys/Gln genotypes,GP chemotherapy showed no significant superior effect in patients with Lys/Lys genotypes,adjusted for gender,age,clinical stage and cytology type(OR=1.09,95%CI:0.95~1.51)-③The incidence rate of vomiting/nausea and alopecie in Lys/Lys genotype was significantly higher than other genotypes(χ2 = 4.032,P = 0.045;χ2 = 4.344,P = 0.037). Conclusion:The polymorphisms of the XPD751 might not be associated with the clinical response to GP chemotherapy in advanced NSCLC,but could be used as a marker of toxicity prognosis.

Abstract:Objective:To investigate the association between of JWA gene single nucleotide polymorphisms and the risk of cervical cancer. Methods:In a hospital-based case-control study of 227 cervical patients and 333 cancer-free controls frequency-matched by age,the genotype of the -76G>C,454C>A and 723T>G polymorphisms of JWA gene were detected by the polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) method;the interaction of menarche age,reproductive age for the first time(RAFT) and parity were further analyzed. Results:A significant increased risk of cervical cancer was found to be associated with the -76GC genotype compared with the -76GG genotype(OR=1.84,95% CI=1.21~2.78),and 454AA genotype compared with the 454CC genotype(OR=1.84,95% CI=1.08~3.14). Moreover,individuals with 3~5 risk alleles will increase the risk of cervical cancer accompanied with early menarche age,late RAFT and many parity. There was no association found between JWA 723T>G polymorphism and cervical cancer. Conclusion:The JWA-76G>C and 454C>A polymorphisms might positively contribute to the etiology of cervical cancer.

Abstract:Objective:To identify the effect of oestrogen receptor(ER) antagonist,Tamoxifen,combination with EGFR tyrosine kinase inhibitor,Gefitinib,on the proliferation and apoptosis in non-small cell lung cancer(NSCLC) cell lines and the related molecular mechanism,providing theoretical evidence for the molecular targeted treatment combination with endocrine treatment in NSCLC. Methods:The cell lines were treated with gefitinib and tamoxifen alone or in combination. The cell proliferation were detected with MTT,the cell apoptosis with flow cytometry and the expression of EGFR and ERβ with real-time PCR and Western blot. In addition,the effect of oestrogen and Tamoxifen on the expression of EGFR,the effect of EGF and Gefitinib on the expression of ERβ were detected,respectively. The effect of oestrogen on the phospho-EGFR were also detected. Results:The combination of Tamoxifen and Gefitinib increased the inhibition and apoptosis in EGFR and ERβ positive cell lines A549 and H1650,while showed no significant changes in EGFR and ER negative cell line H520. It was found that oestrogen could promote the phospho-EGFR protein expression,meanwhile,EGFR protein expression was down-regulated in response to estrogen and up-regulated in response to Tamoxifen in vitro. Conversely,ERβ expression was decreased in response to EGF and increased in response to Gefitinib. Conclusion:The combination of Tamoxifen and Gefitinib can promote its antiproliferative effects on EGFR and ERβ positive lung cell lines. These results suggested that this might be involved in cross-signaling between the EGFR/ER pathways in NSCLC. In conclusion,it provided the rationale to combine Gefitinib with hormone therapy for lung cancer treatment.

Abstract:Objective:To study the antitumor efficiency of Tetrandrine and Cisplatin on transplanted hepatocellular carcinoma(HCC) in mice. Methods:The model of transplanted HCC was established in mice. The tumor volume and weight changes of mice were observed. Results:Tetrandrine could inhibit the growth of the tumor. The inhibition rates were 40.1% and 47.2%, when concentration of Tetrandrine were 5 mg/kg and 10 mg/kg,respectively. The inhibition rates in the two groups were significantly different from that in the control group(P < 0.05). The inhibition rates were 55.8% and 59.9% when the concentration of Cisplatin were 2.5 mg/kg and 5 mg/kg,respectively;however,when combined with 5 mg/kg Tetradrine,the inhibition rates were 73.6%(P = 0.07) and 81.9%(P < 0.05), respectively;when combined with 10 mg/kg Tetrandrine,the inhibition rates were 82.3%(P = 0.06) and 91.4%(P < 0.01). The enhanced anti-cancer efficacy of Cisplatin by Tetradrine was observed in tumor-bearing mice,and had no obvious toxicity reaction. Conclusion:Both Tetrandrine and Cisplatin could inhibit the growth of the transplanted HCC in mice. Tetrandrine could enhance the antitumor effects of Cisplatin. Tetrandrine has synergistic effect with Cisplatin in the treatment of cancer.

Abstract:Objevtive:To explore the gene expressive changes and related genes associated with colorectal cancer and hepatic metastases with gene microarray. Methods:The fresh specimens of primary cancer lesion and hepatic metastasis were collected in 5 patients suffered from colorectal cancer with liver metastasis during operation. After the total RNA extraction,the synthesized biotinylated cRNAs were hybridized to Agilent human 1A oligo microarray. After the design and synthesis of the primers,10 of the differentially expressed genes were then chosen at random to confirm the array results using the SYBR Green Real-time RT-PCR method in 21 fresh specimens collected from primary cancer lesion and hepatic metastasis.The data analysis depended on the ABI Prism 7500 system. Results:Using an extrance limit of ≥2.0 or ≤0.5,there were 110 up-regulated and 96 down-regulated genes identified by microarray which were commonly differentially expressed among the 5 cases. The array results were further confirmed by the real-time RT-PCR. Conclusion:Many genes were involved in the colorectal cancer with liver metastases. It could be helpful to find the regularity of the gene expression by gene microarray and to study the molecular mechanism in the colorectal cancer with liver metastases.

Abstract:Objective:To study the effect of scorpion polypeptide purified from Buthus martensi Karsch(BmK) venom on membrane potential of PC12 cells and its correlation with sodium channel. Methods:PC12 cells were labeled with DiBAC4(3). Laser scanning confocal microscope was used to monitor the changes of membrane potential at real time on these cells that were treated with different concentrations of the scorpion polypeptide. The effect of sodium channel blocker TTX(tetrodotoxin) on the changes was also observed. Results:Membrane potential depolarization induced by the scorpion polypeptide peaked at 3 min and became stabilized after 5 min of the treatment. After comparing to fluorescence intensity without treatment,the normalized fluorescence intensity was determined to be 1.375 ± 0.048,1.504 ± 0.034 and 1.839 ± 0.022 in cells that were treated with 10,20,40 μg/ml scorpion polypeptide,respectively. These numbers are significantly lower than those from untreated control cells(P < 0.01). When PC12 cells were co-incubated with 1 μmol/L TTX for 20 min,then treated with 40 μg/ml scorpion polypeptide,the intensity changed to 1.035 ± 0.028,which was similar to that of the control(P > 0.05). This indicated that the effect of scorpion polypeptide could be completely inhibited by TTX. Conclusion:The scorpion polypeptide induced membrane depolarization of PC12 cells in a concentration-dependent manner,suggesting that the sodium channel should be involved.

Abstract:Objective:To investigate the effect of vasopressin(AVP) on the gastric emptying function in a rat model of psychological stress. Methods:Rats were received intracerebroventriculary injection(icv) with either normal saline or AVP V1 receptor antagonist, and some animal received intravenous injection(iv) of AVP V1 receptor antagonist. Ten min later,animals were subjected to a session of water avoidance stress for a period of 4 h,as previously described. Control animals were maintained in a similar cage without water avoidance for 4 h. The content of cortisone and ACTH in plasma were measured by radioimmunoassay(RIA). Gastric emptying was determined by the phenol red method. Results:Compared with control animals,the levels of ACTH and cortisone in plasma in water avoidance treated rats were increased. Water-avoidance stress stimulated gastric emptying,compared with those without exposure to stress. Intracerebroventricular injection of AVP V1 receptor antagonist could significantly reduce ACTH and cortisone content in plasma. Rate of gastric emptying were also reduced significantly. Conclusion:The enhancement of the hypothalamic-pituitary-adrenal axis function might be essential for the increased gastric emptying rate. These effects might be involved to the increased release of AVP.

Abstract:Objective:To investigate the effects and mechanism of hemeoxygenase-1(HO-1) on acute liver transplantation rejection model. Methods:36 male DA rats used as donors were randomly divided into 3 groups:Group A was injected via vena dorsalis penis with 0.9%NaCl(5 ml/kg);Group B was injected via vena dorsalis penis with cobalt-protoporphyrin(CoPP) 5 ml/kg,hemeoxygenase-1 inducer;and Group C was injected via vena dorsalis penis with zinc-protoporphyrin(ZnPP) (5 ml/kg),hemeoxygenase-1 inhibitor,before livers were harvested. After transplantation,recipients were given the above treatments daily. Six recipients in each group underwent collection of blood and liver tissues 7 days after transplantation to detect the changes of serum enzymes[alanine aminotransferase(ALT),total bilirubin(TBIL)] and the grafts histology. The mRNA and protein expressions of HO-1 and Foxp3 in the transplanted livers were detected by RT-PCR. The other 6 recipients in each group were used to observe the survival time. Results:The survival time of Group B[(29.83 ± 1.40)d] was significantly longer than that of Group A[(13.00 ± 0.52)d,P < 0.01] and Group C[(11.67 ± 0.42)d,P < 0.01]. The values of ALT and TBIL of Group B were significantly lower than those of Group A and C(all P < 0.01). The histological manifestation of the grafts rejection in group A and C were more severe than group B. The expressions of HO-1 mRNA,Foxp3 mRNA in the transplanted livers of Group B were significantly higher than those of Group A. Both mRNA and protein expressions of HO-1 and Foxp3 of Group C were not significantly different from those of Group A. Conclusion:Continuous expression of HO-1 alleviated the reaction of acute rejection in liver transplantation. This effect partly depends on HO-1-mediated Foxp3 activation.

Abstract:Objective:To investigate the early effects of ischemic preconditioning(IPC) on expressions of IKKβ,NF--资B,TNF-α mRNA and the pathological changes in rat kidney allograft tissue. Methods:Thirty-six male SD rats were served as donors and recipients. Animals were randomly divided into the following three groups:the sham-operated group(A), regular transplantation group (B),and treatment group(C). Animals in A group were subjected to exploratory laparotomy. B group received orthotopic homotransplantations. IPC in C group was administered with 15 minutes of ischemic followed by 10 minutes of reperfusion. The changes of renal function and the degree of ischemic reperfusion injury of kidney graft were observed 24 hours after renal transplantation. Expressions of TNF-α,IKKβ,NF-kB p65 subunit mRNA were assessed by RT-PCR,respectively. Results:The degree of renal graft tubules injury in C group was significantly lower than that in the B group(P < 0.01),as well as the transcription of TNF-α(vs non-IPC group P < 0.01) after 24 hours of transplant. No significant difference of the expression of IKKβ mRNA and p65 mRNA was found between C group and B group(P > 0.05). The levels of Scr and BUN were higher than that in A group, whereas no significant difference was found between C group and B group(P > 0.05). Conclusion:One cycle schedule of preconditioning attenuated rat renal allograft ischemia reperfusion injury in early phase. The inhibition of TNF-α expression and the positive feedback of TNF-α/NF-kB signaling pathways might play the role for the protection.

Abstract:Objective:To investigate the role and mechanism of G-protein-coupled receptor kinase interacting protein 1(GIT1) and Extracellular signal-regulated kinases 1 and 2(ERK1/2) in reperfusion of ischemic spinal cord. Methods:The morphology of spinal cord was observed by electron microscope in different time courses after ischemia/reperfusion injury[group A(control group),group B(ischemia 30 min/reperfusion 30 min),group C(ischemia 30 min/reperfusion 2 h),group D(ischemia 30 min/reperfusion 8 h)]. The phosphorylation of ERK1/2 and GIT1 was detected by Western blot. The interaction between GIT1 and pERK1/2 was detected by coimmunoprecipitation. The localization of pERK1/2 was analyzed by immunohistochemistry. Results:The apoptosis of neurocytes of spinal cord undergoing ischemia/reperfusion injury in group D was detected by electronmicroscope. The phosphorylation of ERK1/2 and GIT1 and the interaction between GIT1 and pERK1/2 significantly increased in group C and D. Immunohistochemistry results showed that the stagnation of pERK1/2 was observed in the cytoplasma of neurocytes in group C and D. Conclusion:The stagnation of pERK1/2 in the cytoplasm might act as apoptosis signals contributing to the apoptosis of neurocytes of spinal cord after ischemia/reperfusion injury,and the interaction between GIT1 and pERK1/2 might cause the stagnation of pERK1/2 in the cytoplasm of neurocytes.

Abstract:Objective:To investigate the effects of congestion preconditioning on acute intestinal venous congestion reperfusion injury in rats. Methods:A total of 30 SD rats were divided into three groups,the acute intestinal venous congestion reperfusion group(CR group),the congestion preconditioning group(group CP) and the sham operation group(group SO). To induce intestinal venous congestion,portal vein was clamped for 45 min,then unclamped for 60 min. In group CP,portal vein was clamped for 5 min,then unclamped for 5 min with one repeat of this procedure before model construction. In group SO,all the procedures were conducted except for clamping portal vein. Portal flow was measured with electromagnetic flowmeter at different time point of congestion and reperfusion. After reperfusion for 60 min,ileum samples were collected for MPO and ICAM-1 determination as well as pathological biopsy. Results:In group CP,it was found that the intestinal mucosa was less damaged than that of group CR with Chiu’s Grade Classification. Meanwhile,intestinal MPO activity and ICAM-1 expression in group CP were also much lower than those in group CR. There was no significant difference in portal flow before the clamping of portal vein among the 3 groups. After reperfusion,the portal flow in group CP significantly increased in compare with that of group CR. Conclusion:Congestion preconditioning could alleviate the injury of acute intestinal venous congestion reperfusion in rats.

Abstract:Objective:To analyze the status of pathogenic bacteria distribution and drug resistance of hospital infection in emergency intensive care unit(EICU),for providing the guidance of clinical rational administration and decreasing the hospital infection. Methods:Pathogenic bacteria were isolated from the patients who suffered from hospital infection in EICU from January of 2006 to June of 2007. K-B slip diffusion method was taken to carry out the sensitive test . Rate of drug resistance of the pathogenic bacteria was analyzed. Results:The results showed that rate of the drug fast is very high. As the bacteria distribution ,the main pathogenic bacteria was the gram-negative bacilli(G-)(65.33 %), the second pathogenic bacteria was the gram-positive cocci(G+)(18.29%). and the mycetes(16.38%) occupied the third. In gram-negative bacilli the most were Pseudomonas aeruginosa,Acinetobacter baumannii,Klebsiella pneumoniae and Escherichia coli;Staph is the main pathogenic bacteria in gram-positive cocci;and in mycetes the main pathogenic bacteria were Candida albicans and Candida glabrate. The detection rate of the extended spectrum β-lactamases(ESBLs) from the Escherichia coli and the Klebsiella pneumoniae was 58.06 % and 30.00%. The detection rate of the meticillin-resistant staphylococci(MRS) was 71.87%. Conclusion:Enhance monitoring on pathogenic bacteria distribution and drug resistance analyses of hospital infection in EICU could benefit for the guide of clinical rational administration,depressing multidrug-resistant bacteria and decreasing hospital infection.

Abstract:Objective:To study the feasibility of detecting free-fetal DNA in maternal plasma by multiplex short tadem repeat(STR) loci amplification. Methods:Plasma total DNA extracted from 10 medium pregnancy women by improved DNA isolation method were detected to analyze genotype by multiplex STR amplification. Then the extraction efficiency and rate of assay was analyzed using DNA-PAGE(polyacrylamide gel electrophoresis) in comparison with the maternal and fetal genome DNA. Results:Of all the 60 loci detected in 10 medium pregnancy women plasma,44 loci have results. Except the 14 loci which maternal and fetal DNA genetypes were completely same,free-fetal DNA in maternal plasma were detected in 18 of 30 STR loci,the rate of detection was 30%. D13S317 and D12S391 have the maximal detection rate. Conclusion:Multiplex STR loci amplification can detect free-fetal specificity genotype in maternal plasma,which may have expectation for noninvasive prenatal diagnosis of certain hereditary diseases.

Abstract:Objection:To investigate the significance of contrast-enhanced ultrasound in quantitative diagnosis of liver cirrhosis. Methods:Rabbits models with liver cirrhosis of different stages were successfully made by feeding them with thioacetamide solution. And a self-comparison method was adopted in the trial. The hepatic artery to vein transit time(HA-VTT) was taken as the parameter for evaluation. Results:The rabbit liver cirrhosis model was successfully constructed by thioacetamide treatment. And the HA-VTT had significant changes in cirrhotic rabbit livers. Conclusion:The thioacetamide should be a good agent to make cirrhosis models. And HA-VTT might be an important approach for the quantitative evaluation of liver cirrhosis.

Abstract:Objective:To study the distribution,location and hemodynamic parameter of the blood vessel of anterolateral thigh flap(ALTF) with Color Doppler Flow Imaging(CDFI). Methods:There were 120 normal volunteers’ cutaneous perforators examined on both thighs. The number,diameter,course,location,and the mean velocity(Vmean) were recorded. All these data were analyzed statistically. Results:The average number of cutaneous perforators was 2.25. They were classified into three types:Ⅰmyocutaneous perforator(65.9%),Ⅱseptocutaneous perforator(30.7%),Ⅲdirect cutaneous perforator(3.4%). 97.1% of the dominant cutaneous perforators were located in the middle third of the thigh. There were no significant difference in the Vmean between left and right dominant cutaneous perforators and source vessels. The result was alike among all age groups. Male’s mean velocity was higher than that of female(P < 0.05). Conclusion:The Color Doppler ultrasound with high resolution could show the distribution and location clearly,assess the quality of the blood flow of ALTF,provid scientific basis for successful grafting and have better prospects in clinic.

Abstract:Objective:To evaluate the protection of Ulinastatin(UTI)on pulmonary tissue during cardiopulmonary bypass(CPB). Methods:Twenty-five patients undergoing open-heart surgery under CPB were evaluated in this study. The patients were randomly divided into two groups:control group and experimental group. The experimental group received Ulinastatin as the way of pulmonary protection. Polymorphonuclear neutrophils(PMN) and platelet(Pt) count,interleukin-6(IL-6),interleukin-10(IL-10),tumor necrosis factor-a (TNF-a),alveolar oxygenation index(OI),airway peak pressure and plateau pressure were recorded perioperatively between two groups. Results:The changes of PMN and Pt count in the right atrium blood were significantly higher than that in the left one on 5 min after aorta declamped in control group(P < 0.05). The plasma levels of IL-6 and TNF-a in experimental group were significantly lower than that of the control group(P < 0.05). IL-10 in experimental group increased compared with control group(P < 0.05). OI,airway peak pressure and plateau pressure in control group was significantly higher(P < 0.05) than that of experimental group. Conclusion:Our results demonstrated that PMN and Pt could aggregate in the pulmonary,and IL-6,TNF-a,IL-10 could play an important role in lung injury in CPB. The Ulinastatin could protect pulmonary during cardiopulmonary bypass.

Abstract:Objective:To evaluate the effects of different interventions for flavum ligament in clinical results and CT outcomes on microendoscopic discectomy(MED). Methods:Ninety-two patients were divided into two groups at random. Both groups were disposed with microendoscopic discectomy,differentiated from each other by flavum ligament operation. Data at 1 day pre-operation,3 weeks,3 months and 1 year post-operation were followed up,respectively. The clinical effects were assessed by Oswestry Disability Index,and the improvement rates were also calculate. CT films taken from 1 year follow-up were graded,and the constitution of different graded cases were analyzed between 2 groups. The Correlation between graded CT films and corresponding improvement rates were analyzed. Results:Both ODI at 1 day pre-operation and improvement rates at 3 weeks post-operation were observed no significant difference(P > 0.05). At 3 months and 1 year post-operation,the improvement rates between two groups had significant difference(P < 0.001). In group A,the improvement rates increased at 3 months post-operation in contrast to 3 weeks post-operation(P > 0.05),which declined 1 year later(P < 0.05). No significant difference between 3 weeks and 1 year post-operation was observed(P > 0.05). In group B,the improvement rates increase continually. There was no significant difference(P > 0.05) between 3 weeks and 3 months post-operation,whereas significant differences were observed between 1 year and 3 months,1 year and 3 weeks(P < 0.05). At 1 year post-operation,the constitution of different graded CT films between two groups had significant difference(P < 0.05). Correlative analysis revealed obvious difference between each graded CT and corresponding improvement rate(P < 0.01). Conclusion:Different interventions for flavum ligament can obviously affect the variances of epidural fibrosis that CT films manifested. There was a negative correlation between fibrosis degrees and clinical effects.

Abstract:Objective:To evaluate the therapeutic effect and indication for the treatment of lumbar disc herniation using intradiscal and paraspinal injection with O2-O3 mixture. Methods:One hundred and sixty-two patients were selected and divided into three groups by their category of the lumbar disc herniation which were verified by CT or MR imaging. For each herniated disc treatment,6~15 ml O2-O3 mixture at 40 ug/ml was injected into intradiscal space, and 10 ml of O2-O3 mixture were also injected into the paravertebral space when the needle withdrawned near the nerve root of the intervertebral forame,then the mixture of compound betamethason(1 ml), lidocaine(0.5 ml) and vitaminB12(0.5 mg) was injected. They were also divided into two groups with or without fibrous annulus,the correlation between fibrous annulus broken and therapeutic effect was studied. In addition,three patients with lumbar disc protrusion accompanied spinal stenosis were performed tentative ozone injection. Results:The patients were followed up from 6 months after treatment. The overall efficacy was 76.6%(141/162). Among them,31 cases(91.17%)in 34 bulging patients and 106 cases(89.07%) in 119 patients protrusion had excellent recovery;4 cases(44.44%)in 9 extrusion patients had significant relief of symptoms;63 cases(87.5%)in 72 patients with intact fibrous annulus and 32 cases(68.05%) in 47 patients with broken fibrous annulus had excellent recovery;There was significant difference in therapeutic effect existed between bulging,protrusion patients and extrusion patients. The same outcome was found between intact fibrous annulus groups and broken fibrous annulus groups. Two cases in three patients whose lumbar disc protrusion accompanied spinal stenosis had excellent recovery. No serious complication occurred. Conclusion:The therapeutic method developed by injecting O2-O3 mixture in lumbar intradiscal and paraspinal space was a safe and effective method for the treatment of the lumbar disc herniation. The therapeutic effect was fine in bulging, protrusion groups and intact fibrous annulus groups. Ozone ablation therapy might be tried in the patient with lumbar disc protrusion accompanied spinal stenosis.