Objective To screen the markers of chromosome 12, which is vacant in the Beijing local standard DB11/T828.3-2011 for genetic quality control of miniature pigs in Beijing, and to study the applicability of the local standard by comparison of two different methods for evaluation of the genetic quality of the same population.Methods According to the literature, we selected four pairs of microsatellite markers of chromosome 12 and studied the polymorphism through monitoring the genetic quality of two populations of China Agricultural University miniature pigs.We screened the highly polymorphic markers of chromosome 12, combined them with the microsatellite primers of the standard 18 pairs of chromosomes to establish the whole chromosome method.We compared and analyzed the applicability of the local standard DB11/T828.3-2011 through monitoring the genetic quality of the same population of miniature pigs with different method.The data were processed and analyzed using software Popgen32.Results All the screened four pairs of microsatellite markers of chromosome 12 were highly polymorphic (PIC>0.5).The local standard showed a chromosome coverage of 94.7%, stability of amplification of 96.0%, and certified that the China Agricultural University miniature pig III was qualified, while the China Agricultural University miniature pig I was not qualified.When the markers of chromosome 12 were added, the whole chromosome method showed result of 100% chromosome coverage and 96.6% amplification stability, both of the two populations of pigs were certified as qualified.Conclusions The four screened markers of chromosome 12 are all highly polymorphic, and provide a support for supplement of the local standard DB11/T828.3-2011.

Objective To strengthen the quality control management and enhance the detection capacity of the experimental animal quality control laboratoriesin our country through the detection of serum esterase-1 (Es-1) in the experimental mice.Methods The samples were prepared according to the standard procedure, and then were randomly numbered and distributed to participating units by cold-chain transport.Before the deadline, the participants submitted the results and the copies of original records.When the results were completely consistent with the standard results,the results were regarded as satisfactory, otherwise were unsatisfactory.Results A total of 11 laboratories participated in this program, of which 10 laboratories were regarded as satisfactory (90.9%) and one laboratory obtained unsatisfactory result (9.1%).Conclusions The results of this proficiency testing project demonstrate that the overall detection level of Es-1 in laboratory mice is highof the participating laboratories.However, more attention still should be paid to standard specifications and some test details.

Objective To investigate the detection capacity of malic enzyme 1 and isocitrate dehydrogenase 1 ( Mod1&Idh1) in the laboratory animal monitoring laboratories in China in order to understand the detection capacity of la-boratories and to improve the detection level of laboratory animals’ quality.Methods Based on the program approved by CNAS, samples preparing, homogeneity test and stability test of malic enzyme 1 and isocitrate dehydrogenase 1 in the mouse kidneys were carried out.Standard operation procedure and samples with random numbers were distributed to the la-boratories.The laboratories should submit the result reports before the time limit expires.If the laboratory reports were the same with the standard results, the laboratories will receive satisfactory remark.If laboratory reports were not the same with the standard results, the laboratories will receive unsatisfactory remark.If a laboratory did not submit report, the laboratory will also receive unsatisfactory result.Results Eight laboratories out of 10 (80%) enrolled laboratories reported satisfac-tory experiment results, and two laboratories (20%) presented unsatisfactory results.Conclusions The whole detection level of laboratories in Mod1 &Idh1 is relatively high in the laboratory animals monitoring laboratories in China.It can re-flect the detection level of laboratories to conduct the laboratory capacity evaluation.

Objective To systematically evaluate the effectiveness and safety of glucoside tripterygium total (GTT) combined with antihistamine medicine for chronic idiopathic urticaria (CIU). Methods All randomized or semi-randomized controlled trials (RCTs or semi-RCTs) of GTT in treating CIU were collected from CNKI, VIP, WanFang Data, CBM, Embase, PubMed and Cochrane Library clinical controlled trials database. After two researchers conducted screening and data extraction independently, the quality of the included literature research was evaluated according to the risk of bias tool described in the Cochrane Handbook version 5.1.0, and then RevMan 5.3 was used to undertake Meta analysis. Results A total of 21 articles about RCTs and semi-RCTs were obtained, including 2194 patients. Meta analysis indicated that GTT combined with antihistamine medicine treatment showed higher effective rates compared with the control group [RR=1.33, 95%CI (1.26, 1.40), P<0.000 01], with obviously lower recurrence rate [RR=0.45, 95%CI (0.33, 0.62), P<0.000 01]. There was no statistical significance in adverse reactions. Conclusion GTT combined with antihistamine medicine for CIU has definite efficacy, and is relatively safe. Whether apply GTT combined with antihistamine medicine for CIU should be verified through randomized controlled trial with large-scale samples, multiple centers and high quality.

Objective To investigate the role of hypoxia?inducible factor?2α(HIF?2α) in the expression of tight junction proteins and permeability alterations in rat glomerular endothelial cells (rGENCs) under hypoxia condition. Methods The expressions of the HIF?2α and tight junction proteins such as occludin and ZO?1 of rGENCs were examined after exposed to 5%oxygen at different treatment time periods (0 h, 12 h, 24 h and 48 h). Then lentiviral transfection was used to knock down HIF?2α expression in rGENCs. The cells were split into four groups, including i) control group where rGENCs were cultured under normal oxygen conditions, ii) hypoxia group, iii) negative control group where rGENCs were infected with a negative vector, iv) HIF?2α lentivirus transfection group. Group ii, iii and iv were kept in hypoxic chamber (5% O2, 5% CO2 and 90% N2) for 24 h. The expressions of occludin, ZO?1 and HIF?2α were assessed by Western blotting. The permeability of rGENCs was measured using trans?epithelium electrical resistant (TEER) by Millicell? ERS voltohmmeter. Results With the elongation of hypoxia time, the expression of HIF?2α was increased gradually, while the occludin expression was decreased, there was statistically significance difference in each group (all P<0.01). The expression of ZO?1 also decreased gradually under hypoxia circumstance, but no statistically significant was found between 24 h and 48 h groups (all P>0.05). And a dramatic decrease in TEER of hypoxia cells was detected as compare with control cells (P<0.01). After knockdown of HIF?2αexpression, both expressions of occludin and ZO?1 were increased significantly compared with hypoxia cells (P<0.01), and TEER elevated at the same time (P<0.01). Above indexes had no statistical difference between hypoxia cells and negative control cells (all P>0.05). Conclusion Hypoxia may promote HIF?2α expression, which could increase the permeability of rGENCs by reducing the expression of occludin and ZO?1.

Objective:To investigate the effect of rapamycin (Rapa) on apoptosis of acute myeloid leukemia THP-1 cells induced by idarubicin (IDA) and its molecular mechanism.Methods:The THP-1 cells were treated with 10, 20, 40 and 80 nmol/L Rapa for 1 h, and the cells without Rapa treatment were set up. Western blot was used to detect the conversion of autophagy marker LC3 protein in THP-1 cells (the ratio of LC3Ⅱ/LC3Ⅰ), flow cytometry was used to detect the apoptotic rate, and the pretreatment concentration of Rapa was determined. THP-1 cells were treated with different concentrations of IDA for 24 h, the cell proliferation inhibition rate of IDA for THP-1 cells was detected by CCK-8 method, and the half maximal inhibitory concentration ( IC50) was calculated. THP-1 cells with or without Rapa treatment were treated by IDA with the concentration of lower than IC50 for 24 h, CCK-8 method was used to detect cell proliferation inhibition rate, flow cytometry was used to detect cell apoptosis, real-time fluorescent quantitative polymerase chain reaction was used to detect the expression changes of autophagy-related genes Beclin-1, LC3 and p62, and Western blot was used to detect the conversion of autophagy marker LC3 protein. Results:The ratio of LC3Ⅱ/LC3Ⅰ in THP-1 cells treated by 20 nmol/L Rapa was higher than that in the untreated cells ( P=0.002 4). The apoptotic rate in THP-1 cells treated by 80 nmol/L Rapa was higher than that in the untreated cells ( P=0.007 3). According to the results of Western blot and flow cytometry, 20 nmol/L Rapa was selected as the pretreatment concentration. The IC50 of IDA for THP-1 cells treated with IDA for 24 h was 59.874 nmol/L. After treated with 50 nmol/L IDA for 24 h, the proliferation inhibitory [(69.67±5.03)% vs. (41.67±3.51)%] and apoptotic rates [(74.35±4.83)% vs. (41.25±5.24)%] in THP-1 cells pretreated by Rapa were higher than those in the unpretreated cells (both P<0.05); the Beclin-1 and LC3 mRNA expression levels and the ratio of LC3Ⅱ/LC3Ⅰ in THP-1 cells pretreated by Rapa were higher than those in the unpretreated cells, and the expression of p62 mRNA was lower than that in the unpretreated cells (all P<0.05). Conclusion:Rapa can enhance the apoptosis of THP-1 cells induced by a relative low dose of IDA, which may be achieved through inducing excessive autophagy in THP-1 cells.

Objective To investigate effects of high fat-salt diet on change of growth and development,body fat distribution insulin sensitivity and associated metabolic indexes for juvenile rats. Methods 50 grams of male,female SD juvenile rats (3 weeks,just weaned) were randomly divided into 3 groups,12-14 animals in each group,were given routine diet (NC) and high fat diet (FC) and high fat-salt diet (FSC) .Then the body weight,,body length,abdominal circumference,blood pressure,visceral fat weight,plasma lipids were measured 4 weeks later,at the same time oral glucose tolerance test and insulin release test were performed. Results In the FSC group,body weight,abdominal circumference,blood pressure,visceral fat,plasma glucose and insulin level significantly increased than the NC group,plasma lipid disorders increased and significant insulin resistance occurred. Conclusion High fat and high salt could successfully induced obesity,hypertension,dyslipidemia and impaired glucose tolerance.

AIM: To investigate the effects of adenovirus-mediated human tissue kallikerin (Ad-hKLK1) gene delivery on the proliferation, migration of VSMC_(SHR) induced by platelet derived growth factor-BB (PDGF-BB). METHODS: The VSMC_(SHR) proliferation induced by PDGF-BB was accessed by cell counting and methyl thiazolyl tetrazoliuin (MTT). The migration was assessed by modified Boyden chamber assay. Western blotting was used to determine the expressions of the cycle-independent kinase inhibitors p27~(Kip1) and p21~(Cip1).RESULTS: Proliferation of VSMC_(SHR) induced by PDGF-BB was inhibited after transfection of Ad-hKLK1 (20-100 MOI) in a MOI-dependent manner. The peak inhibition titer of Ad-hKLK1 fell on 100 MOI, with the peak inhibition rate of 39.3% (cell counting, n=3, P<0.01), 30.2% (MTT, n=3, P<0.01), peak stunning rate of cell-cycle in phase G0/G1 at 36.4%. The inhibitory effects of proliferation and cell-cycle caused by hKLK1 gene delivery were significantly abolished by Hoe140, a bradykinin B2 receptor antagonist. Migration of VSMC_(SHR) induced by PDGF-BB was inhibited after hKLK1 gene delivery, with the peak inhibitory rate of 34.6% (n=6, P<0.01). However the inhibitory effects of migration were not blocked by Hoe140. The protein expression of p27~(Kip1) and p21~(Cip1) increased significantly after the hKLK1 gene delivery, whereas Hoe140 nearly completely blocked these effects (n=3, P<0.01, respectively).CONCLUSION: The hKLK1 gene delivery may inhibit the proliferation and migration of VSMC_(SHR) induced by PDGF-BB. Bradykinin B2 receptor probably mediates the up-regulating expression of p27~(Kip1) and p21~(Cip1) that contributes to the inhibitory effects of proliferation of hKLK1. However, the inhibitory effects of migration by hKLK1 gene delivery may not be mediated by bradykinin B2 receptor.

Objective To investigate the pattern of antihypertensive medication prescribing in outpatients from the Second Hospital of Tianjin Medical University, and analyze the shortcoming and deficiency compared with 2010 Chinese guidelines for the management of hypertension. Methods A total of 154 262 electronic prescribing for outpatients with hy-pertension, from January-December 2012 in a Grade 3A hospital in Tianjin, were enrolled in this retrospective survey. Data of commonly used antihypertensive medication and combination therapy in patients were analyzed. The patient data collected were divided into different groups according to age, gender, high blood pressure level and the onset of the season. Results (1)The list of the drugs commonly used for treating hypertension in outpatients were calcium antagonist (52.3%), angiotensin receptor blockers (34.0%),βblockers (25.9%), angiotensin-converting enzyme inhibitors (12.1%), fixed-dose combination (11.0%) and diuretics (1.4%).(2)The fewer combination therapy was found in outpatients than that of monotherapy (43.9%vs 56.1%). Some prescriptions were not routinely recommended by the Guideline (4.6%).(3)The combination therapy used in patients with stage 3 hypertension was higher than that of patients with stage 1or stage 2 hypertension (44.5%vs 37.7%vs 37.7%, P<0.01). The rate of combination therapy was significantly higher in cardiology department than that of other clini-cal departments (P<0.01). The combination therapy tended to be used in the elderly patients than that of non-elderly pa-tients (P<0.01). The number of prescriptions was lower in summer than that of other seasons,but the rate of combination therapy was higher in summer than that of spring, autumn and winter (P<0.01). Conclusion The prescriptions of combina-tion therapy and diuretic were inadequate in outpatients with hypertension. These findings indicate the difference between clinical prescription and the guideline for the management of hypertension.

Objective To explore the efficacy and safety of percutaneous transhepatic portal vein or transjugular intrahepatic portosystemie shunt (TIPS)to implant the portal vein metallic stent in treatment of cavernous transformation of portal vein (CTPV).Methods Clinical and imaging data of 8 patients with CTPV were retrospectively analyzed who were treated in our hospital.All patients were treated with metallic stent implantation in portal vein including 3 patients by TIPS and 5 by percutaneous transhepatic portal vein.Results All patients were successful in the stent implantation without any occurrence of serious complications such as intra-abdominal hemorrhage and so on.Intraoperative angiography showed blood circulated freely in these stents.1 day-2 weeks later,the patients symptoms of abdominal pain and gastrointestinal bleeding were obviously relieved or disappeared.Follow up 1 month-3 years,1 patient with stent occlusion after one year of operation,the blood flow recovery after stent reimplantation,and the remaining patients,color doppler ultrasound reflected patency of blood flows in their stents.No one suffered from gastrointestinal bleeding or abdominal pain again.Conclusion Implantation of portal vein metallic stent via percutaneous transhepatic portal vein or via TIPS in treatment of cavernous transformation of portal vein is safe and effective.