Objective To study the effective methods for extraction of clenbuterol from biological material. Methods Clenbuterol in blood samples was extracted with 5 different methods,the extracts from various extraction methods were determined by gas chromatography-mass spectrometry(GC-MS?雪. The efficiency of 5 extraction methods was compared with each other. Results The standard curve showed good linear relationship with a range from 0.01-10.0 mg/L clenbuterol hydrochloride(r=0.999 9?雪. The detection limit was 0.005 mg/L. After adding 1.0 ?g/ml clenbuterol hydrochloride into blood sample,the blood sample was immersed and extracted in 0.01 mol/L HCl and absolute alcohol.The extracts were extracted by chloroform at pH value of 11. The average recovery and RSD of this method were 96.5% and 5.21% respectively(n=6?雪. Conclusion The method showed easy operation and reliable results ,it was suitable for determination of clenbuterol in biological material without deviation.

Objective To determine whether tissue factor(TF) is present in the amniotic fluid of normal term pregnancy. Methods Tissue factor antigen levels in amniotic fluid, blood plasma, centrifugal supernatant and sediment of 16 normal term pregnancies were detected with immunoenzymatic method(ELISA). Results Tissue factor antigen levels in the blood plasma, the whole amniotic fluid, the centrifugal supernatant and the sediment were [(31?9) ng/L0, [(404?186) ng/L], [(348?177) ng/L] and [(1360?639) ng/L], respectively. Tissue factor antigen concentrations in three states of the amniotic sediment(P

Objective Analyze the relationship between high sensitivity C‐reactive protein (hs‐CRP) ,homocysteine (Hcy) ,D‐di‐mer (D‐D) levels and atherosclerosis (AS) .Methods A total of 92 patients with AS were analyzed retrospectively ,including acute coronary syndrome (ACS) group with 61 cases ,stable angina pectoris (SAP) group with 31 cases .According to the result of coro‐nary artery plaque CT detection was further divided into plaque group ,non plaque group ,stable plaque subgroup ,and unstable plaque subgroup .Meanwhile 42 healthy subjects were selected as control group .The levels of the three indicators were compared . Results The serum hs‐CRP ,Hcy and plasma D‐D concentration in the ACS group and SAP group were significant higher than those of the control group (P< 0 .05) ,and those of the ACS group were significant higher than those of the SAP group(P< 0 .05) . The serum hs‐CRP ,Hcy and plasma D‐D concentration in the plaque group were significant higher than those of the non plaque group ,those in the unstable plaque subgroup were significant higher than those of the stable plaque subgroup(P< 0 .05) .The serum hs‐CRP ,Hcy and plasma D‐D concentration of the ACS patients were all correlated positively .Conclusion Serum hs‐CRP ,Hcy and plasma D‐D levels are closely related to the development of AS ,the combined detection of three indexes is value for the prevention , treatment and prognosis of AS .

Objective To investigate the changes of the activity of blood coagulation and fibrinolysis in fracture patients,and to research the relations between the activity and thrombossis. Methods This reser ch chose 60 patients in the department of orthopaedic trauma in our hospital and 10 normal control.Each patient enderwent laboratory analysis on preoperation?post operation 24 hours?post operation 4 days.The molecule markers of coagulation and fibrinolysis TAT(thrombin/antithrombin Ⅲ complex)?F1+2(prothrombin fragment F1+2)?PAP(Plasmin/? 2-antiplasmin complex) were quantitative measured by enzyme immunoassay. Result Compare with normal control,F1+2 and TAT levels increase obviously(P0.05); FG levels increase obviously (P0.05) .Conclusion It has a high level of the activity of blood coagulation and has no obviously change of the activity of fibrinolysis after trauma injury,it indicates high coagulation activity is correlated with thrombossis.

Objective To investigate the effect of neutrophil extracellular traps (NETs) on inflammation of rheumatoid arthritis (RA), especially angiogenesis. Methods The presence of NETs in synovial tissues of RA and osteoarthritis (OA) was observed by immunofluorescence assay. Neutrophils were isolated from peripheral blood of health volunteers. Neutrophils were cultured in vitro, the formation of NETs was observed. NETs were extracted as a stimulating agent. The effects of NETs on the proliferation of human umbilical vein endothelial cells (HUVECs) and synovial fibroblasts (RAFLS) were evaluated by MTT, and which were classified into two groups: HUVECs group and RAFLS group, with the following treatment: control and NETs (0.28 mg/L). Wound repair assay was employed to evaluate the effect on the cell migration stimulated with NETs. The experiment was divided into three groups:control, VEGF (40μg/L VEGF) and NETs (0.28 mg/L NETs). Results (1) Compared with OA, NETs were found more in the synovial tissue of RA. (2) NETs formation was induced by stimulator in vitro. The concentration of extracted NETs-DNA was 27.8 mg/L. (3) MTT assay showed that compared with the control groups, low concentration of NETs (0.28 mg/L) promoted the proliferation of HUVECs (0.499 ± 0.011 vs. 0.393 ± 0.009, P<0.05) and RAFLS (0.266 ± 0.007 vs. 0.192 ± 0.007, P<0.05). (4) It was showed that a significant wound closure induced by low concentration of NETs (0.28 mg/L) was found compared with control. Conclusion Our present study suggests that NETs are found more in the synovial tissue of RA, and low concentration of NETs can promote angiogenesis in RA.

Objective To investigate the neutrophil extracellular traps (NETs) formation and their molecular mechanisms induced by serum amyloid A (SAA) in rheumatoid arthritis (RA).Methods Neutrophils were isolated from peripheral blood of RA and healthy volunteers.① Neutrophils were cultured in vitro,the formation of NETs was observed and their percentage was calculated.② Neutrophils were cultured in vitro,divided into six groups:control,SAA,[SAA+anti-Toll like receptro4 (TLR4)-Ab],LPS,(LPS+anti-TLR4-Ab) and anti-TLR4-Ab.Appropriate stimulation was conducted for each group.NETs formation and their percentages were investigated.The concentration of DNA in supernatant was detected by fluorescent staining.F test and t test were used for statistical analysis.Results ① The purification of isolated neutrophils was higher than 95％.The network which was collocated with the spreading neutrophils nucleus and neutrophil elastase under the microscope,was NETs.In the RA group,the formation of NETs induced by SAA was significantly more than control [(19.1±0.8) vs (7.4±0.5),t=12.30,P＜0.05].② However,after pretreated with anti-TLR4 antibody,NETs formation was significantly less than the SAA group [(5.7±0.4) vs (14.7±1.1),t=7.825,P＜0.05].Moreover,the fluorescence intensity of DNA in supernatant was significantly higher in SAA group than that of anti-TLR4-Ab pretreatment group [(18.7 ±0.7) vs (12.9±0.8),t=5.552,P＜0.05].The concentration of DNA in supernatant of SAA group was higher than that of anti-TLR4-Ab pretreatment group [(36.9±1.3) μg/ml and (16.3±0.6) μ,g/ml,t=14.41,P＜0.05].Conclusion SAA can induce the formation of NETs from neutrophils by binding to TLR4 in RA.

Objective T o explore the relationship betw een the change rules of volatile organic com pounds (V O C s) in rat m uscle and postm ortem interval (PM I). Methods A total of 120 healthy rats w ere divided random ly into 12 groups (10 for each group). A fter the rats w ere sacrificed by cervical dislocation, the bodies w ere kept at (25±1)℃. R at m uscle sam ples w ere separately obtained at 12 PM I points, including 0, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10 d. T he V O C s in rat m uscles w ere collected, detected and ana-lyzed by headspace solid-phase m icroextraction (H S-SPM E ) coupled to gas chrom atography-m ass spec-trom eter (G C-M S ). Results In total, 15 species of V O C s w ere identified, including 9 arom atic com-pounds, 3 sulfur com pounds, 2 aliphatic acids and 1 heterocyclic com pound. T he species of V O C s in-creased w ith PM I: no species w ere detected w ithin 1 day, 3 species w ere detected on day 2, 9 on day 3, 11 on day 4, 14 from day 5 to 7, and 15 from day 8 to 10. T otal peak area of 15 species of V O C s w as significantly correlated to PM I (adjusted R2=0.15-0.96): the regression function w as y=-17.05 x2+164.36 x-246.36 (adjusted R2=0.96) from day 2 to 5, and y=2.24 x+101.13 (adjusted R2=0.97) from day 6 to 10. Conclusion T he change rules of V O C s in rat m uscle are helpful for PM I estim ation.

Objective To observe the efficacy and safety of pigolitazone/metformin fixed-dose combination therapy replacing metformin alone or combined with other anti-diabetes drugs in type 2 diabetes with poor glycemic control.Methods 80 cases were recruited,with an average age of (54.79±13.99)years,diabetes history of (9.76±6.59) and baseline HbA1c (9.06±1.34)％.All participants received pigolitazone/metformin instead of metformin without other treatment changes.Glycemic control (level of fast blood glucose,HbA1c) was evaluated at 12 weeks,as well as lipid profiles,liver and renal function,adverse events and body weight.Results 8 cases were lost to visit,4 cases were withdrawn for edema,only 68 subjects finished the study.Compared to the baseline,after 12-week treatment,FPG decreased for (2.06+0.16) mmol/L,HbA1c decreased for (0.84+0.23)％,both of the differences were statistically significant (P＜0.001,P＜0.001).Body weight increased (0.34+1.13)kg,with no difference compared to the baseline.The lipid profile presented elevated high density lipoprotein cholesterol (P=0.012)and decreased total cholesterol,low density lipoprotein cholesterol,triglyceride,while the latter three items showed no differences (P＞0.05,P＞0.05,P＞0.05).Indexes reflecting liver and renal function,such as ALT,AST,TBIL,DBIL,Urea,UA,Cr showed no differences compared with the baseline.Adverse events analysis showed at the end of the study,no severe hypoglycemia and serious cardiovascular events occurred,6 cases suffered edema,among whom 4 patients exited the study for severe lower limb edema.No extra gastrointestinal symptom happened.Conclusion Pigolitazone/metformin fixed-dose combination exhibits an excellent efficacy and safety for T2DM,with satisfying tolerability and compliance,which is a selection for those patients with poor glycemic control.

Objective:To construct the pET30a-EgG1Y162-2 prokaryotic expression plasmid and induce the expression of EgG1Y162-2 protein, so as to provide a research basis for development of Echinococcus granulosus vaccine. Methods:Using Echinococcus granulosus cDNA as a template, the target gene of EgG1Y162-2 was synthesized by PCR, and after digestion with restriction enzymes EcoRⅠ and Hind Ⅲ, it was connected to the prokaryotic expression vector pET30a to construct the recombinant plasmid pET30a-EgG1Y162-2. The recombinant plasmid was transformed into competent cell BL21 (DE3) and induced by isopropyl β-D-thiogalactoside (IPTG) to express a large number of proteins. The recombinant protein was purified by affinity chromatography. The purification level was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the expression product was identified by Western blotting. Results:The recombinant plasmid pET30a-EgG1Y162-2 was successfully constructed. After inducting expression, the bacterial supernatant and the eluate were both at a relative molecular weight of about 15 × 10 3, and the protein antigen component eluted with 200 mmol/L imidazole was relatively pure. Western blotting results showed that the purified recombinant protein EgG1Y162-2 with His tag could be recognized by His monoclonal antibody. Conclusion:The pET30a-EgG1Y162-2 prokaryotic expression plasmid of Echinococcus granulosus is successfully constructed, and the recombinant protein of EgG1Y162-2 is induced to express, laying a foundation for further study on anti- Echinococcus granulosus vaccine.

Objective To investigate college students' awareness and cognition of 84 shared courses on the Internet and analyze the discrepancies between different ethnicity and majors in Xinjiang Medical University. Methods Purposive sampling was adopted with questionnaire of 1 448 students of 26 classes in Xinjiang Medical University. The survey aimed to investigate students' awareness and cognition of these In- ternet courses. Obtained data were statistically analyzed using SPSS 13.0, and the test level α=0.05. Result The hits of preventative medicine, Chinese medicine, clinical medicine and pharmacy were 19.0%, 33.3%, 35.7% and 11.9% respectively. Among the courses whose hits exceeded 50,000, clinical medicine accounted for 50%, preventive medicine for 21.4%, Chinese medicine and pharmacy for 14.3% respectively, there being no significant difference in overall hits (P>0.05). 90.1% of the surveyed students knew shared courses on the Internet, there being no difference between gender, ethnicity and profession (P>0.05). 89.8% had an accurate understanding of the concept of shared courses on the Internet, there being differences between gender and major ( χ2gender=11.013, P=0.026; χ2major=136.08, P=0.000) without significant differences in ethnicity ( χ2=11.378, P=0.497). 71.9% of the students used shared courses on the Internet as reference resources for the course study, 46.6% for the learning content, 35.5%for the understanding of other resources related to the course, 30.1% for lab-class and exercises, 10% for discussion on online course forum. The Han and Kazak students used the courses as a study, experiment and exercise tool, while the Uyghur and Kazak mainly used these for learning and discussion on the forum, there being differences in ethnicity ( χ2=26.889, P=0.001); the usage rate of the courses of preventive medicine students is higher, whereas pharmacy students relatively low, there being significant differences in major ( χ2=38.01, P=0.004). The Kazakh and Uygur students mainly used the courses to formulate learning plans, and the Han students to learn the current curriculum. 44.7% of preventive medicine students used the courses to improve their abilities, which was significantly higher than those of other majors. Only 18.4% and 1% of pharmacy students used the courses to formulate learning plans and to improve their abilities, which were both lower than those of other majors, showing ethnic and professional differences ( χ2ethnic=37.654, P=0.001; χ2major=73.68, P=0.000). Conclusion Students' awareness of shared courses on the Internet is high and their cognition is accurate. However, there are differences in the ways and purposes of the employment of the courses between different ethnicity and majors. The main reason may be related to major, the quality of the courses, as well as the lack of effec-tive supervision and evaluation system, suggesting that the management department in our university should strengthen the supervision and evaluation of the courses and give full play to the important role of shared courses on the Internet.