A LAMP-Taqman rapid detection system for porcine pseudorabies virus(PRV)was de-veloped based on LAMP and quantitative PCR.LAMP primers were designed for PRV conserved sequences,and the loop primer modified by the fluorescent quenching group was used as the Taq-man probe.The composition optimization,specificity,sensitivity and repeatability of the LAMP-Taqman system were tested using positive samples and recombinant plasmid as templates.Thirty-eight samples of pork swabs were tested with the commercial LAMP detection kit in parallel to verify the actual detection effect of the LAMP-Taqman detection system.The results showed that the optimal final concentration of each component was as follows:PRV-FIP/BIP 0.8 μmol/L,Bst DNA polymerase 0.7 U/μL,Taq DNA polymerase 0.24 U/μL,dNTPs 1.6 mmol/L,MgSO47.2 mmol/L.This system had good specificity and did not cross-react with other virus samples.The linear correlation coefficient of gradient samples was 0.995,the coefficient of variation of repeatable tests was less than 3.000％,and the minimum detection limit could reach 2.81 ×102 copies/μL.The test results of the actual swab samples were consistent with the commercial isothermal fluores-cence detection reagents.In conclusion,the PRV detection system established by LAMP-Taqman method in this study is specific,sensitive,stable and accurate,and is a reliable technical method suitable for the accurate detection of porcine PRV.

Objective:This paper summarized the practical experience in the construction of a scientific research platform in a large comprehensive hospital and discusses the innovative management model of the scientific research platform.Methods:This paper introduced the development history and construction status of a scientific research platform in a large comprehensive hospital, and analyzes the contradictions and difficulties existing in equipment sharing, resource allocation and other aspects during the development of the scientific research platform and its countermeasures.Results:Building a strong scientific research platform was of great significance to the hospital's discipline development, talent cultivation, and improvement of academic influence and core competitiveness. The hospital should continue to increase investment in the construction of scientific research platforms and strengthen system layout, cooperation, exchanges, and resource sharing.Conclusions:Large comprehensive hospitals need to build a highly open and efficient scientific research platform system to empower technological innovation.

OBJECTIVE To research the effect and mechanism of fermentation of Astragalus membranaceus on endogenous metabolites in hyperuricemia model rats using serum UHPLC-HRMS. METHODS The SD rats were randomly divided into different groups, including blank group, model group, benzbromarone group(20 mg·kg-1), as well as fermentation of Astragalus membranaceus high-dose(3 g·kg-1) and low-dose group(1.5 g·kg-1). Model group and each treatment group were disposed with 300 mg·kg-1 oxonic acid potassium to establish hyperuricemia models. At the time of 1 h after modeling, rats in each treatment group were given corresponding drugs for intervention. Collected rat serum after 14 d. The serum of different groups were collected for endogenous metabolites research using UHPLC-HRMS. After multivariate statistical analysis, the different metabolites and metabolic pathways were selected. RESULTS The hyperuricemia rat modes were successfully established by oxonic acid potassium 14 d, and fermentation of Astragalus membranaceus showed good uric acid reducing effect. Compared with the blank group, 17 potential biomarkers associated with hyperuricemia were found in the model group. Among them, 9 potential biomarkers were significantly recalled by fermentation of Astragalus membranaceus. It mainly involved sphingolipid metabolism, pyrimidine metabolism, tryptophan metabolism, pantothenic acid and CoA biosynthesis, glycine, serine and threonine metabolism and other pathway. CONCLUSION This study can provide a basis for revealing the mechanism of reducing uric acid by fermentation of Astragalus membranaceus, and lay a foundation for the further development and utilization of Astragalus.

Objective To investigate the expression of pseudokinase Tribbles homology 3(TRIB3)and its clinical prognostic value in Siewert type Ⅱ adenocarcinoma of esophagogastric junction(AEG).Methods Western blot and immunohistochemical method were used to detect the expression of TRIB3 in R0 resected Siewert type Ⅱ AEG and its corresponding adjacent tissues,and analyze its rela-tionship with clinical parameters,survival and prognosis.Results Western blot analysis showed that the expression level of TRIB3 in Siewert type Ⅱ AEG tissues was significantly lower than that in the adjacent tissues(P<0.05).The immunohistochemical Results showed that the positive expression rate of TRIB3 in cancer tissues was significantly lower than that in adjacent tissues(P<0.01).The expression of TRIB3 was significantly correlated with the degree of differentiation,clinical TNM stage and lymph node metastasis(P<0.05),but not with age,gender and pathological morphology(P>0.05).Kaplan-Meier survival analysis showed that the long-term survival of patients with positive TRIB3 expression was significantly better than that of patients with negative TRIB3 expression(P<0.01).Univariate(HR =0.290,95%CI:0.110-0.761,P =0.012)and multivariate(HR =0.179,95%CI:0.051-0.630,P = 0.007)COX regression analysis showed that TRIB3 could be used as an independent prognostic factor for patients with Siewert type ⅡAEG(P<0.05).Conclusion TRIB3 may be involved in the occurrence and development of Siewert typeⅡ AEG.It is expected to be-come a new target for early diagnosis and treatment of AEG,and can be used as an important indicator for judging the prognosis of patients.

Objective:To determine the risk profile of venous thromboembolism (VTE) and evaluate VTE prophylaxis implementation of the hospitalized cancer patients in the DissolVE 2 study.Methods:The data of hospitalized cancer patients in the DissolVE 2 study were analyzed. The risk distribution of VTE, preventive measures and in-hospital VTE events of hospitalized patients with tumors were described by percentage and 95% confident interval (CI).Results:A total of 1 535 cancer patients were included. According to the Padua score, 826 (53.8%) patients were at low risk of VTE, while 709 (46.2%) patients were at high VTE risk. VTE events occurred in 4 low-risk patients (0.5%; 95% CI: 0.1%, 1.2%) and 5 high-risk patients (0.7%; 95% CI: 0.2%, 1.6%). The overall incidence was 0.6% (9/1 535, 95% CI: 0.3%, 1.1%). Among patients with high VTE risk, 666 (93.9%) did not receive any VTE prophylaxis, and only 11 (1.6%) patients received appropriate VTE prophylaxis. Among patients who received VTE prevention, no VTE event was observed. Conclusions:Nearly half of the hospitalized cancer patients are at high risk of VTE, but most of them don′t receive VTE prophylaxis. The results reflect the insufficient management of VTE risk for hospitalized cancer patients in China, and improvement of awareness and practice of VTE prophylaxis is urgently needed.

Objective:To determine the risk profile of venous thromboembolism (VTE) and evaluate VTE prophylaxis implementation of the hospitalized cancer patients in the DissolVE 2 study.Methods:The data of hospitalized cancer patients in the DissolVE 2 study were analyzed. The risk distribution of VTE, preventive measures and in-hospital VTE events of hospitalized patients with tumors were described by percentage and 95% confident interval (CI).Results:A total of 1 535 cancer patients were included. According to the Padua score, 826 (53.8%) patients were at low risk of VTE, while 709 (46.2%) patients were at high VTE risk. VTE events occurred in 4 low-risk patients (0.5%; 95% CI: 0.1%, 1.2%) and 5 high-risk patients (0.7%; 95% CI: 0.2%, 1.6%). The overall incidence was 0.6% (9/1 535, 95% CI: 0.3%, 1.1%). Among patients with high VTE risk, 666 (93.9%) did not receive any VTE prophylaxis, and only 11 (1.6%) patients received appropriate VTE prophylaxis. Among patients who received VTE prevention, no VTE event was observed. Conclusions:Nearly half of the hospitalized cancer patients are at high risk of VTE, but most of them don′t receive VTE prophylaxis. The results reflect the insufficient management of VTE risk for hospitalized cancer patients in China, and improvement of awareness and practice of VTE prophylaxis is urgently needed.

Objective:To assess the antibody persistence 3-5 years following vaccination of measles and rubella combined live-attenuated vaccine (MR) at 8 months of age and measles, mumps and rubella combined attenuated live vaccine (MMR) at 18 months of age.Methods:In 2016, 18-month-old children who were vaccinated with one dose of MR vaccine at the age of 8 months were recruited in Hebei Province as group 1; 4-, 5- and 6-year-old children who were vaccinated with one dose of MR vaccine at the age of 8 months and one dose of MMR vaccine at 18 months of age were recruited in Shanxi, Inner Mongolia and Beijing as group 2, group 3 and group 4, respectively. Serum samples were collected to detect IgG antibodies against measles, mumps and rubella by ELISA. Geometric mean concentrations (GMCs) of measles, mumps, and rubella antibodies were compared among groups by analysis of variance or non-parametric test. Seropositive rates were compared among groups by Chi-square test or Fisher′s exact test. Results:A total of 650 children were included in this study. Seropositive rates of measles, mumps and rubella antibodies 30 d after vaccination of 150 18-month-old children with one dose of MMR vaccine were 100%, 91.33% and 100%, respectively, and the GMCs were 1 846.87 mIU/ml, 299.91 IU/ml and 111.33 IU/ml, respectively. Seropositive rates of measles, mumps and rubella antibodies 3-5 years after vaccination one dose of MR vaccine at 8 months of age and one dose of MMR vaccine at 18 months of age were above 94%, 79% and 71%, respectively, and the GMCs were above 830 mIU/ml, 240 IU/ml and 31 IU/ml. No significant difference in the seropositive rates of the three antibodies was observed among groups 2, 3 and 4 ( P>0.05). There was no significant difference in the GMCs of measles or mumps antibodies among the three groups ( P>0.05), but the differences in the GMCs of rubella antibodies were statistically significant ( P=0.034). Conclusions:Measles, mumps and rubella antibodies persisted for 3-5 years without significant decrease after vaccination one dose of MR vaccine at 8 months of age and one dose of MMR vaccine at 18 months of age.

Objective To evaluate the safety and efficacy of retroperitoneal laparoscopic procedure for rad-ical nephrectomy. Methods The clinical data of 140 cases underwent retroperitoneal aparoscopic radical nephrec-tomy from August 2013 to December 2015 were retrospectively analyzed. Among them ,87 cases were controlled by procedure for radical nephrectomy ,53 cases were treated with Non-procedural for radical nephrectomy. The peri-operative information parameters of the two groups were compared and analyzed. Results All of the 140 cases were operated under laparoscopy. There was no significant difference between the two groups in the basic condition and tumor location(P>0.05). Retroperitoneal laparoscopic procedure in the operation time ,intraoperative bleed-ing,peritoneal rupture rate,blood transfusion,complete tumor resection and postoperative drainage were better than non-procedural for radical nephrectomy ,the differences were statistically significant(P<0.01). All patients were followed up for 1 to 3 years without recurrence and metastasis. Conclusion Retroperitoneal laparoscopic pro-cedure for radical nephrectomy is significant ,worthy of clinical application and promotion.

Objective To study the antitumor effects on human prostate cancer cell lines of a traditional Chinese medicine named atractylodes macrocephala koidz volatile oil in vitro.Methods LNCaP and DU145 cell lines were normally cultured and were divided into 4 control groups including:Blank culture without serum and without cell (group A),Blank culture with serum but without cell (group B),LNCaP cell culture with serum (group C),DU145 cell culture with serum (group D).In the meanwhile,there were 6 experimental groups:adding 50 μg/ml of atractylodes macrocephala koidz volatile oil in culture of group C1 and group D1,250ug/ml in group C2 and group D2,and 500ug/ml in group C3 and group D3.All of the 10 groups were simultaneously cultured in 24-well-plates for 48 hrs,and each group was repeatedly studied for three times.Forty-eight hours later,every 2 × 106 cells of LNCaP or DU145 was seeded into each well and atractylodes macrocephala koidz volatile oil was added in 6 experimental groups,and saline water was added into 4 control groups.Another 48 hrs later,the culture solutions of the 10 groups were separately collected for testosterone (T),estradiol (E2),progesterone (P),vascular endothelial growth factor (VEGF),basic fibroblast growth factor (b-FGF),total prostate specific antigen (tPSA) and free prostate specific antigen (fPSA) analysis with enzyme-linked immunoassay kits.The experiment was repeated for 3 times,and the mean data were statistically analyzed by SPSS One-way ANOVA.Results The growth of cultured cells was found to have been effectively inhibited in group C1 and group D2.The inhibition severity of LNCaP cells was positively related with the drug concentration and time,while DU145 cells could only be highly inhibited (60.96％) after 24hrs drug treatments.In group C and group D,we found that both T were in very low level (0 ng/ml) whereas both E2 were in high levels (269 pg/ml and 239.81 pg/ml,P ＜ 0.05),no distinct differences showed in P; In addition,VEGF,b-FGF and fPSA were all in high values,whose values were 102.96 pg/ml and 1763.40 pg/ml,0.26 ng/ml and 6.41 ng/ml,0.16 ng/ml and 0.44 ng/ml,respectively; DU145 cells had higher values than LNCaP cells (P ＜ 0.05).As regard to the 6 experimental groups,in the groups C1,C2,C3 and D3,we found that T had been unexpectedly increased from 0 to 0.37 ng/ml (P ＜ 0.05),E2 continuously elevated from 239.81 pg/ml to 649.90 pg/ml (P ＜0.05),surprisingly P were also increased from 0.98 ng/ml to 9.83 ng/ml (P ＜0.01).On the contrary,VEGF,b-FGF and fPSA levels were all graduallydecreased,dropping down to 47.79 pg/ml and 59.56pg/ml,0 and 1.79 ng/ml,0 and 0.11 ng/ml,respectively; nevertheless,in group C2 and group D2,fPSA values were surprisingly increased from 0 and 0.04 ng/ml up to 1.78 ng/ml and 0.23 ng/ml respectively.Conclusions Atractylodes macrocephala koidz volatile oil has certain anti-tumor effects on human prostate cancer.Compared with LNCaP cells,DU145 cells have many different characteristics in sex hormone,cytokine and PSA expressions.

Objective To investigate the feasibility and accuracy of volumetric measurement of necrotic lesion using CT and MRI,and to assess the value of necrotic lesion volume in predicting collapse of the femoral head in patients with avascular necrosis of the femoral head(ANFH). Methods Comparison among CT,MRI and gross section was performed in 25 femoral heads of 18 patients who underwent total hip replacement for established ANFH.The volume of necrotic lesion was measured using fluid displacement.CT and MRI data were transferred to a computer to calculate the volume of necrotic lesion using software.One way ANOVA was used to compare the volumes of necrotic lesion measured by CT,MRI and gross section.A total of 62 patients (92 hips) who were diagnosed with ANFH but without collapse by CT were followed up 24 months.Student t-test was used to compare the ratio of the volumes of the necrotic lesion and entire femoral head in the hips with and without collapse and ROC curve analysis was carried out.Results CT and MRI coincided with gross section in the necrotic area,proliferative area and extralesional area.The volumes of the necrotic lesion measured by CT,MRI and gross section were ( 20.5 ± 5.2 ),( 21.4 ± 4.8 ),( 20.9 ± 5.2 ) cm3,respectively.There was no significant difference among the necrotic volumes measured by the three methods(F =0.185,P =0.831 ).In fifty-seven out of 92 hips,collapse of the femoral head occurred during the follow-up.The ratio of the volumes of the necrotic lesion and entire femoral head was higher in hips with collapse than in hips without collapse[ (34.5 ±9.3)％ vs.(23.4 ±8.4)％ ;t =5.749,P=0.000].The area under the ROC curve was 0.808. Conclusions The volume of the necrotic lesion plays an important role in the collapse of femoral head in patients with ANFH.Both CT and MRI can identify the shape and location of the necrotic lesion intuitively and stereospecifically and can determine the volume of the necrotic lesion accurately.