At present, there are many methods for genotype of hepatitis C virus , but not a gold standard. In order to establish the rationale for genotypic determination of optimal region sequence, fifteen complete genome sequences of hepatitis C which had been given the annotation about every region and derived from different country were downloaded from GenBank. Phylogenetic trees on 5' UTR, core, E1, E2 and NS5B region were established. The results demonstrated that genotyping group was not all correct on 5' UTR region while genotyping groups were wholly correct on core, E1, E2 and NS5B region. Comparing phylogenetic distances on core, E1, E2 and NS5B region with that on complete genome sequence demonstrated that the NS5B area was the best genotyping region instead of the complete genome sequence. In addition, analysis of the molecular evolution on each core region could supply some clues for creating novel genotyping method based on PCR-RFLP.

Objective To study basic thyroid stimulating hormone (bTSH) levels impact on outcomes of in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) in Qinghai.Methods Totally 282 cases with IVF cycles and 93 cases with ICSI cycles were studied prospectively,according to bTSH level,patients were divided into four groups.Reproduction rate,clinical pregnancy rate,miscarriage rate and live birth rate were studied among four groups.Results (1) In 375 cases with IVF/ICSI cycles,bTSH was positively correlated with abortion rate (r=0.42,P=0.04),but live birth rate and growing rate showed negative correlations with bTSH (r=-0.42,-0.28; P=0.04,0.03).bTSH and the number of eggs,the number of fertilized eggs,the number of embryos,biochemical pregnancy rate,and clinical pregnancy rate were no significant correlation (all P＞0.05).(2) Among women at group of ≤1.7,＞1.7 and ≤2.5,＞2.5 and ≤3.5,＞3.5 mU/L,the implantation rates were 28.7％,27.3％,37.7％ and 19.2％,live birth rates were 80.9％,75.0％,82.7％,and 59.8％,abortion rates were 19.0％,15.0％,16.7％,40.1％; they all showed significant difference (all P＜0.05).Abortion rate in women with high bTSH level was higher than that of women with lower bTSH level,however implantation rate,live birth rate in women with high bTSH level were lower.Conclusion When bTSH level is ＞3.5 mU/L,the abortion rate were increased,but live birth rate,rate of implantation were decreased.

In this article, the authors expounded the necessary and problems of animal experiment post-ap-proval monitoring ( PAM) , which included who should do PAM, how to do PAM, and what the PAM can do. The authors also exposed the following suggestion: formulating the detailed rules, regulations and SOP, strengthening the training of PAM team member and animal experiment personnel, and monitoring the whole process of animal protocol review using the software.

Objective To study the effect of cyclophosphamide(CP) and its metabolites acrolein(ACR) on PTEN gene deleted on chromosome 10 after acting on ovarian cancer cellsSKOV3.Methods Different concentrations of CP and ACR were selected to act on recombinant PTEN protein.The phosphorylation activity of PTEN was detected by PNPP.The expression of PTEN protein was detected by Western blot.The binding mode of drug with protein was detected by the biotin combined with protein;meanwhile the expression change of P53/TP53 in PTEN gene pathway was analyzed.The target protein was obtained by immunoprecipitation(IP) after different drug concentrations acting on the cells.The phosphorylation activity of the target protein was detected by high performance liquid chromatography(HPLC).Results After the drug metabolites acting on recombinant PTEN protein,the phosphorylation activity was decreased with the increase of drug concentration,while the expression of ACR antibody action was increased with the drug concentration elevation.The expression of protein and biotin in different experimental groups was increased with the increase of drug concentration.The PTEN phosphorylation activity was decreased with the drug concentration increased in cells,and so did the expression of TP53 protein.Conclusion CP metabolite ACR induces the cytotoxicity by inhibiting PTEN protein phosphorylation activity.

Objective To improve the diagnosis and treatment of infective endocardits(IE)by exploring the causes,pathogenic microorganism,clinical characteristics and therapeutic results of patients with IE.Methods From January 2006 to January 2016,89 cases with IE in Ganzhou People's Hospital were collected,and the clinical charac-teristics were analyzed retrospectively.Results Among the IE patients,40.5%had congenital heart diseases,33.7%had rheumatic heart diseases,18.0% were post－PCI or operative endocarditis,and patients without basic cardiac diseases accounted for 7.9%.The most common clinical manifestations were fever (86.5%),followed by anemia (85.4%).The blood culture positive rate was 63.4%.Streptococci viridians were the most common causative organ-isms(44.2%),followed by coagulase －negative staphylococci(21.2%).Eighty－six patients received echocardio-graphy examination,vegetation was observed in 61cases (68.5%).The hospital mortality rate was 10.1%,mostly due to refractory congestive heart failure and sepsis.Conclusion Congenital heart disease showed an upward trend, rheumatic heart disease decreased significantly.Intermittent fever,anemia are the main clinical manifestations.Blood culture positive rate is not high,Streptococcus viridians is the most common causative organism.Echocardiography is the main basis for the diagnosis.

Cardiovascular diseases have become the most common and most prevalent chronic diseases in the elderly.Smoking, hyperlipidemia, diabetes and other high-risk factors participate in the occurrence and development of atherosclerosis by impairing the structure and function of the vascular wall, resulting in the occurrence of arterial stiffness.Pulse wave conduction velocity(PWV) is the rate at which a pulse wave travels from one specific location of the artery to another specific location along the wall of the artery.It is a simple, effective and repeatable noninvasive index for evaluating arterial elasticity and vascular function.PWV is closely related to cardiovascular disease.In recent years, the relationship between PWV and cardiovascular disease has been deeply studied.The relationship between PWV and coronary heart disease, hypertension, cardiac function, atrial fibrillation and pulmonary hypertension is described in this paper.

Objective:To analyze the fail mode of neoadjuvant therapy combined with surgery for locally advanced esophageal squamous cell carcinoma (ESCC) after long-term follow-up.Methods:Clinical data of consecutive 238 patients with locally advanced resectable ESCC who underwent neoadjuvant therapy combined with surgery in Zhejiang Cancer Hospital from September 2012 to October 2019 were retrospectively analyzed. The failure mode in the whole cohort was analyzed after long-term follow-up. The overall survival (OS) and disease free survival (DFS) rates were analyzed by Kaplan-Meier method. Survival differences were determined by log-rank test.Results:The pathological complete response (pCR) rate was 42.0% in 238 patients. After a median follow-up of 46.1 months, tumor progression occurred in 96 patients (40.3%), including 25 patients (10.5%) with local recurrence, 61 patients (25.6%) with distant metastases, and 10 patients (4.2%) with simultaneous local recurrence and distant metastases. The median OS and DFS were 64.7 months and 49.9 months. And the 3-, 5-, and 7-year OS and DFS rates were 70.0%, 52.8%, 36.4% and 63.5%, 42.5%, and 30.0%, respectively. The 3-, 5-, and 7-year locoregional recurrence-free survival rates and distant metastasis-free survival rates were 86.0%, 71.4%, 61.2% and 70.6%, 55.9%, 43.0%. Compared with non-pCR patients, the overall progression rate and distant metastasis rate of pCR patients were lower (26.0% vs. 50.7%, 16.0% vs. 32.6%, both P<0.05). And the 3-, 5-, and 7-year OS (83.0% vs. 60.2%, 69.7% vs. 41.7%, 50.4% vs. 27.7%, all P<0.001) and DFS rates (80.4% vs. 51.4%, 63.9% vs. 31.2%, 45.9% vs. 20.3%, all P<0.001) were significantly better in pCR patients. Conclusions:Distant metastasis is the main failure mode of patients with locally advanced ESCC after neoadjuvant therapy. Patients with postoperative pCR can achieve better long-term survival.