OBJECTIVE To detect and analyze the results and significance of HBV-DNA,HBeAg and PreS1 of HBV infective sera.METHODS Routine detection and PreS1 antigen of 450 sera were tested by ELISA,and HBV-DNA was detected by fluorescent quantitation PCR.RESULTS The positive rates of HBV-DNA,HBeAg and PreS1 were 74.4%,48.9% and 63.3%,respectively,in 450 cases of HBV infective sera.Among 285 PreS1-positive samples,the positive cases of HBeAg and HBV-DNA were 190 and 270,respectively.There were significant difference(P0.05)among PreS1,HBV-DNA and PreS1,and they three were in positive correlation.CONCLUSIONS PreS1 and HBV-DNA are more sensitive than HBeAg,and PreS1 is better coincided with HBV-DNA.They can reflect the infection and replication condition of HBV.Therefore,it has important clinical meaning for the diagnosis and therapy of HBV to simultaneously used HBV-DNA,HBeAg and PreS1.

Objective Using chemically synthesized small interfering RNA (siRNA) transfected HepG2.2.15 cells to construct a cell model in interfering hepatitis B virus (HBV) X gene, studying the inhibi-tion of HBV replication and antigen expression in vitro. Methods After transfection of HepG2.2.15 cell for 24 h, 48 h, 72 h, detecting the cell supernatant of HBsAg and HBeAg by chemiluminescence immunoassay, the cell supernatant HBxAg protein by ELISA , the HBx mRNA relative expression of transfected cell was detected by fluorescence quantitative polymerase chain reaction (PCR), the ability of cell proliferation was detected by CCK-8 assay. Results After HBx-siRNA transfected HepG2.2.15 cells, cell proliferation ability was inhibited. The cell of HBx mRNA and the cell supernatant of HBxAg expression decreased (P < 0.05); at the same time it in-hibited the expression of HBsAg and HBeAg. The suppressed peak and the inhibition rate were 66% and 58%respectively at 72 h. The fluorescence quantitative PCR confirmed that expression of HBV DNA in the super-natant was decreased. Conclusion The HepG2.2.15 cell interference model of HBV X gene has been success-fully constructed, which has an effect of inhibiting proliferation of HepG2.2.15 cells and replication and expres-sion of HBV gene in vitro.

Five groups were assigned to include intrahepatic cholangiocarcinoma ( ICC, n=30 ) , liver cirrhosis (LC,n=30),metastatic carcinoma (MCA,n=30) and 30 healthy subjects.The serum level of GPC3 was measured by a sandwich method of enzyme-linked immunosorbent assay ( ELISA ) and alpha-fetoprotein (AFP) by microparticle enzyme immunoassay (MEIA).The serum levels of GPC3 and AFP were significantly higher than those of other groups (P<0.05).At a cut-off value of 3.5μg/L,the sensitivity and specificity of GPC3 in the diagnosis of HCC was 83.3%and 76.7%respectively.The sensitivity of combined measurement of GPC3 and AFP was better than GPC3 or AFP alone.Detectable GPC3 was significantly correlated with the presence of viral hepatitis markers and tumor size.However there was no obvious difference in tumor thrombi in portal vein ( PVTT), tumor number, age, gender or hepatic function of HCC.Thus,as a sensitive serum diagnostic marker for HCC ,GPC3 may be a good supplement to AFP in differentiating HCC from non-malignant chronic liver diseases and other liver cancers.

[ ABSTRACT] AIM:To investigate the effect of phycocyanin on the apoptosis of human laryngeal cancer HEP-2 cells and to explore the inhibitory mechanism of phycocyanin to tumor.METHODS:Highly purified phycocyanin was ex-tracted from spirulina.The effects of phycocyanin at different concentrations on the growth of human laryngeal cancer HEP-2 cells were detected by MTT assay.In addition, the cell structures were observed under electron microscope.The cell ap-optosis was analyzed by flow cytometry.The production of reactive oxygen species ( ROS) was measured by flow cytometry. Enzymatic activities of caspase-3,-8 and-9 were measured by chemical colorimatry.The expression of Bax, Bcl-2, Fas, P53, caspase-3 and caspase-9 at mRNA and protein levels was determined by RT-PCR and Western blot.RESULTS:MTT test confirmed that phycocyanin inhibited the cell activity of HEP-2 cells with time and dose dependent manners.The result of electron microscope observation and flow cytometry indicated that phycocyanin induced the apoptosis of HEP-2 cells.The intracellular content of ROS was increased.The activities of caspase-3, -8 and -9 were increased.RT-PCR showed that the mRNA expression of Bax, Fas, P53, caspase-3, caspase-9 was increased and Bcl-2 was decreased.The results of Western blot were consistent with the results of RT-PCR.CONCLUSION:Phycocyanin might induce apoptosis of HEP-2 cells by down-regulating Bcl-2, up-regulating Bax, Fas and P53, and the transduction of apoptotic signals in the human laryngeal cancer cells.

Education of evidence-based medicine in clinical research is important in enhancing primary hospital clinical research capacity and quality.Dongyang People's Hospital of Zhejiang Province has carried out evidence-based medicine education and training,and greatly improved the performance of clinical research and development of scientific researchers.

Due to the insufficient education of medical ethics and tending to medical moral educa-tion, the residents lack the ability to make ethical decisions. Therefore, we applied Four Topics method in the clinical ethics training of residents. The clinical ethics training was divided into three stages, and each stage had different training content and objectives. Dongyang People's Hospital adopted the form of lectures and panel discussions. During the group discussion, the group leader was in charge of case preparation, which came from clinical practice, and then all members applied the Four Topics method to analyze and discuss the ethical conflicts, and make the ethical decision. Through this process, residents' ability to solve ethical problems in clinical practice was enhanced.

Clinical thinking of Clinicians in Dongyang People's Hospital is gradually shifting from ‘empirical medicine’ to ‘evidence-based medicine’ after popularizing of evidence-based medicine, training core members and implementing multi-layered, multi-channeled, comprehensive and sustainable resident doctor training mode with clear steps and purposes within three years. This paper discussed on reasonable clinical practice of evidence-based medicine in primary hospital and provided realistic basis for the further development and improvement of training of evidence-based medicine in primary hospital.

Objective:Our study aims to predict acute kidney injury (AKI) in acute myocardial infarction (AMI) by establishing a random forest model.Methods:By using the clinical database from affiliated Dongyang Hospital of Wenzhou Medical University, a total of 1 363 AMI cases were included. Then, 75% of participants were analyzed as training subsets and the remaining 25% were testing subsets. The CARET package in R was used to filter variables and build random forest. The prediction ability of established model was evaluated by specificity, sensitivity, accuracy, relative operating characteristic curve (ROC curve) in testing subsets. In addition, the performance of random forest was compared with other 3 commonly used machine learning algorithms (Artificial Neural Network, Naive Bayes, and Support Vector Machine).Results:In this study, 30 variables including the demographic information, risk factors of cardiovascular disease, vital signs at admission, laboratory tests were identified and used to establish our random forest prediction model. The area under the curve of the testing subsets ROC was 0.893. The specificity and sensitivity of prediction model was 0.791 and 0.866, respectively. And the first creatinine, first blood urea nitrogen, and D-dimer after admission, age, mechanical ventilation were the top-five factors in this model. After comparing various machine learning algorithms, random forest model had a better performance.Conclusion:The random forest model would be used to predict the occurrence of AMI with AKI.

Objective To investigate the expression of CCL2 in colorectal cancer and its carcinogenesis mechanism associated with macrophages.Methods The expression level of CCL2 mRNA in 17 cases of colorectal cancer tissues and corresponding adjacent tissues were analyzed by PCR,and the phenotypes of macrophages in tumor infiltrating lymphocytes (TIL) were analyzed by flow cytometry.Human peripheral blood mononuclear cells were isolated and induced to differentiate into macrophages.After 12 h incubation with CCL2,the phenotypic changes of macrophages were analyzed by flow cytometry.The expression level of VEGF,COX-2 and IL-6 in the supernatant were measured by ELISA assay.Results The expression level of CCL2 in colorectal cancer tissues was significantly higher than that in corresponding adjacent mucosal tissues (t =4.017,P ＜ 0.05),and the macrophages in TIL had a high proportion of CCR2 phenotype.CCL2 was shown to induce increased CCR2 expression in macrophages (t =5.070,P ＜ 0.05),and promote the secretion of the tumor growth-associated factors such as VEGF,COX-2 and IL-6 (all P ＜ 0.05).Conclusion The levels of CCL2 in colorectal cancer were up-regulated suggesting that CCL2 may play a key role in tumor promotion by recruiting macrophages and influencing their function.

Objective To discuss the clinical application of mechanical thrombectomy with AngioJet system for acute lower limb arterial ischemia (ALI).Methods A total of 12 ALI patients,who underwent percutaneous mechanical thrombectomy with AngioJet systemn during the period from January 2015 to November 2016,were enrolled in this study.The clinical data were retrospectively analyzed.The blood flow classification score after thrombolysis in myocardial infarction (TIMI) was used to evaluate the blood perfusion condition,and Cooley standard of efficacy score was used to assess the clinical curative effect.Results The technical success rate of mechanical thrombectomy with AngioJet system was 91.7％ (11/12).The average restored perfusion time was (1.5±0.6) hours.The clinical success rate and limb salvage rate were 83.3％ (10/ 12) and 91.7％ (11/12),respectively.The TIMI flow scores were improved from preoperative grade 0 (n=8) and Ⅰ (n=4) to postoperative grade 0 (n=1),Ⅰ (n=3) and Ⅱ (n=8).In 11 patients (91.7％) the symptoms of lower limb arterial ischemia were strikingly improved after mechanical thrombectomy.In one patient,the postoperative TIMI flow score remained 0 as preoperative state and the symptoms of lower limb arterial ischemia were not improved although catheter directed thrombolysis therapy was employed for 24 hours,and above-knee surgical amputation had to be carried out.Cooley efficacy score showed that complete cure was seen in 4 patients (33.3％),good response in 6 patients (50.0％),general improvement in one patient (8.3％) and pool response in one patient (8.3％).No severe bleeding complications occurred.Conclusion Percutaneous mechanical thrombectomy with AngioJet can rapidly recover the blood perfusion in patients with ALI,thus,further deterioration of the disease can be prevented and the limb salvage rate can be improved.Therefore,this technique has good clinical application value.