Objective To construct a subtractive cDNA library of genes transactivated by XTP3 protein with suppression subtractive hybridization technique in order to clone genes associated with transactivation. Methods mRNA was isolated from HepG2 cells transfected by pcDNA3.1(-)-XTP3 and pcDNA3.1(-) empty vector respectively, and then cDNA was synthesized. After restriction enzyme Rsa Ⅰ digestion, small sizes cDNAs were obtained. Then tester cDNA was divided into two groups and ligated to the specific adaptor 1 and adaptor 2 respectively. After being hybridized with driver cDNA twice and underwent two times of nested PCR, tester cDNA was subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E.Coli strain JM109. The cDNA was sequenced and analyzed in GenBank with Blast search after PCR. Results The amplified library contained 30 positive clones. Colony PCR showed that 23 clones contained 200-1 000bp inserts. Sequence analysis was performed also. It was found that 20 kinds of known and 2 kinds of novel cDNA sequences may be target genes transactivated by XTP3 protein. Conclusion The subtractive library of genes transactivated by XTP3 protein was constructed successfully. It provides a foundation for the further research on pathogenesis of the viral proteins.

Objective To investigate the risk factors and treatment of postoperative delirium in patients with benign prostatic hyperplasia(BPH). Methods　For 256 postoperrative BPH patients in our department,the clinical parameters including age,hypoxemia,preoperative infection,postoperative pain,and introvert were analyzed,and the treatment were summarized. Results　The incidence of postoperative delirium was higher in patients aged≥75 years than＜75 years[17.4%(19/109)vs.5.4%(8/147),X2=8.307,P=0.0043,higher in patients with preoperative infection (pulmonary or urinary)versus without the infection[15.0%(17/113)vs.7.0%(10/143),X2=4.337,P=0.037],higher in patients with hypoxia versus without hipoxia[16.5%(16/97).vs.6.9%(111159),X2=4.885,P=0.027]and higher in patients with post-operative pain versus without the pain(16.9%(10/59)vs.1.2%(1/85),X2=10.146,P=0.0014].Hematocrit and hemoglobin in patients with postoperative delirium were lower after operation versus pre-operation[(0.29±0.04)vs.(0.42±0.04),t=12.2314,P＜0.001;(102.39±6.58)g/L vs.(114.58±8.36)g/L,t=5.9537,P＜0.001].All delirium patients were recovered after sedation treatment,improving the quality of sleep,controlling the infection and correcting electrolyte turbulence. Conclusions　Advanced age,hypoxemia,preoperative infection,postoperative pain are important risk factors for delirium.

Objective To establish the determination method of Tubeimoside Ⅰ in Jiazhongxiao preparation and serum of rat. Methods RP-HPLC/UV method was used as follows:Diamonsil C18 column (250 mm?4.6 mm, 5 ?m) and Easyguard column, methanol-water (65∶35) as mobile phase with flow rate of 1.0 mL/min, UV detection at 214 nm. Results The mean recovery rates of Tubeimoside Ⅰ in preparation and serum of rat were (103.49?3.49)% and (104.70?4.69)% respectively. The linearity of Tubeimoside Ⅰ was shown in the range of 1.18～88.5 ?g/mL. Tubeimoside Ⅰ in preparation and serum of rat were 995.3 ?g/g and 3.19 ?g/mL. Conclusion The method was simple, reliable and rapid. It’s quite suitable to be used in the analysis of Tubeimoside Ⅰ in Jiazhongxiao preparation and serum of rat.

Objective To evaluate the influences of artifical plasma substitutes on blood coagulation and fibrinoglysisMethods Forty adult patients, ASA grade Ⅰ-Ⅱ, scheduled for selective surgery ,were randomly allocated to receiving intravenous infusion of hydroxyethylh starch(HES group), polygeline (Haemaccel,HAE group), succinylated gelatin (Gelofusine,GEL group), or normal saline (control group) at the rate of 20ml?kg -1?h -1in 60 min ,respectively, with 10 cases in each groupThe venous blood samples were taken before and 1h following the infusion to determine platelet count(PLC),platelet agglutgtination test(PAG), platelet factor 3 availability test(PF 3at), activated partial thromboplastin time(APTT), prothrombin time(PT), thrombin time(TT), fibrinogen(FIB), tissue plasminogen(t-PA) and plasminogen activator inhibitor(PAI:A)Results In HAE group PAG decreased significantly, and PT was prolonged markedly after the infusion(P005)In GEL group only FIB reduced obviously after the infusion ,additionally was remarkably lower than that in control group(P

Objective To investigate effects of intrathecal injection of sufentanil combined with bupivacaine on postoperative analgesia and immunity function in elderly patients under lower extremity orthopaedic surgery.Methods Totally 60 elderly patients with ASA Ⅰ or Ⅲ under lower extremity orthopaedic surgery were randomly divided into two groups:bupivacaine and sufentanil group (each n =30).3 ml bupivacaine (2 ml 0.75％ bupivacaine plus 1 ml 0.9％ normal saline) were given in bupivacaine after successful spinal anesthesia and 3 ml sufentanil (1 ml 0.75 ％ bupivacaine plus 1 ml 0.9％ normal saline and 1 ml with 5 μg sufentanil) were used in sufentanil group respectively.The anesthetic feeling and block level before operation recovery time were recorded postoperation.Visual analogue scale (VAS) and adverse reaction were recorded at 1,6,12,24 and 48 h after surgery.Cellular and humoral immunity function were recorded at the following time points:T0,the day before surgery; T1,the day after surgery; T2,the first 5 postoperative days; T3,the first 10 postoperative days; T4,the first 15 postoperative days.Results The sensory and motor block levels in sufentanil group were T9～～T10 and T11～T12 respectively,which were higher than in bupivacaine (T7～T8 and T8～T10).The recovery time in sufentanil group was shorter than in the bupivacaine (P＜0.05).There were no significant differences in VAS scale between the two groups at any postoperative time points (P＜0.05).The incidence of skin pruritus was higher in sufentanil group (6 cases) than in bupivacaine (P＜0.05).There were significant differences in the indexes for the immune condition including CD3+,CD4+/CD3+,CD4/HLADR,CD8/HLADR,B-cell,NK-cell between time points of T2[(48.7±2.3)％,(33.4±1.8)％,(10.2±1.8)％,(0.75±0.07)％,(1.02±0.16)％,(3.12 ±0.19) ％,(2.53±0.14)％,respectively] and T0 in bupivacaine (P＜0.05).There were significant differences in the indexes for the immune condition including CD3+,CD4+/CD3+,CD4/HLADR,CD8/HLADR,B-cell,NK-cell at T2 between sufentanil group [(54.5 ± 2.4) ％,(36.3 ± 1.7) ％,(15.1±1.6)％,(1.32±0.11)％,(1.25±0.14)％,(4.96±0.25)％,(4.17±0.10)％,respectively] and bupivacaine (P＜0.05).Conclusions Intrathecal injection of 5 μg sufentanil combined with low concentration bupivacaine can be safely and effectively applied in elderly patients under lower extremity orthopaedic surgery,and can reduce local anesthetic concentration,shorten recovery time of anesthesia and accelerate the recovery of postoperative immunity function.

Objective To explore the characteristics and risk factors of type 2 diabetes mellitus (T2DM) onset in pedigrees. Methods A total of 865 subjects were screened and diagnosed by oral glucose tolerance test (OGTT) based on American Diabetes Association (ADA) criteria. Type 1 diabetes mellitus (T1DM) , maturity onset diabetes of the young (MODY) and chondriosome diabetes were excluded by clinical features and laboratory test of insulin and autoantibodies including glutamic acid decarboxylase antibody, insular cellular antibody and insulin autoantibody. A total of 182 pedigrees of T2DM were obtained. Results No gender difference was found in the prevalence of T2DM (42. 59% in male and 48. 18% in female respectively, P ＞0. 05) , nor was the newly diagnosed rate(9. 89% in male and 11. 82%in female, P ＞ 0. 05). The onset age was (63. 3 ± 12. 4) years old in the first generation [(64. 4 ± 12. 5)years in male and (62. 3 ± 10. 3) years in female] , (47. 1 ± 8. 7) years old in the second generation [(48. 2 ±9. 3)years in male and (46. 1 ± 8. 1) years in female] , (29. 6 ± 10. 2) years old in the third generation [(28. 9 ±9. 5)years in male and (30. 0 ± 10. 4)years in female]. Compared with normal glucose tolerance (NGT) subjects , newly diagnosed T2DM and impaired glucose regulation (IGR) subjects had higher prevalence of hypertension, hyperlipidemia and smoking but less physical activities. Statistical differences were shown in body weight five years before diagnosis, one years before diagnosis and at diagnosis in newly diagnosed T2DM[(68. 4 ±12. 4)kg, (69. 5 ± 11. 0)kg and (69. 1 ±9. 6)kg] and IGR[(66. 1 ±10.7)kg, (65.9 ± 10.7) kg and(65.7 ± 10.4) kg] , when compared with NGT [(61.0 ± 10.2) kg,(59. 5 ±11.0) kg and (60. 1 ± 10. 4) kg, all P ＜ 0. 05] . The same results were obtained with waist circumference and waist-hip ratio [(4. 1 ± 12. 5) cm and 0. 92 ± 0. 36 in newly diagnosed T2DM while (89. 1 ± 10. 7) cm and 0. 90 ± 0. 64 in IGR] , when compared with NGT[(82. 5 ± 10. 1) cm and 0. 82 ±0. 25] , all P ＜0. 05. Conclusions No gender difference was found in the onset characteristics of T2DM.High prevalence of obesity, hypertension, hyperlipidemia and smoking with less physical activities were associated with T2DM.

Objective To study risk factors of cardiovascular disease (CVD) and status of bone mineral density (BMD) in women with hypoestrogenism.Methods From Jul 2011 to April 2013,a total of 256 women with hypoestrogenism in the First Affiliated Hospital of Shanxi Medical University were enrolled in this retrospective study,which were divided into four groups:133 women in ppausal group,25 women in premature ovarian failure (POF) group,67 women in menopausal transition group and 31 women in premature ovarian failure transition group.General statue,CVD risk factors and BMD were compared among four groups.General statue include menopausal period,menopausal symptoms (Kupperman Index),CVD risk factors include body mass index,blood pressure,waist circumference,waist-hip ratio,blood lipids and glucose,BMD include left hip,lumbar spine bone mineral density and T or Z value.Results (1) The median menopausal period were 3.4 years in postmenopausal group and 3.6 years in premature ovarian failure group,which did not show no statistical difference (P ＞ 0.05).Kupperman Index in four groups were 12 in postmenopausal group,9 in POF group,9 in menopausal transition group and 8 in premature ovarian failure transition group,which reached statistical difference (P ＜ 0.05).(2) The difference of body mass index (BMI),waist circumference,waist-hip ratio,diastolic blood pressure were no statistically significant among four groups(P ＞ 0.05) ; the systolic blood pressure in four groups were 120,110,110,110 mm Hg (1 mm Hg =0.133 kPa),their differences were statistically significance (P ＜ 0.05); the high-density lipoprotein (HDL-C) was 1.6 mmol/L in postmenopausal group,and 1.3 mmol/L in premature ovarian failure transition group,their differences were all statistically significance (P ＜ 0.05) ; the difference of the fasting plasma glucose (FPG) was not statistically different in 4 groups (P ＞0.05).(3) The abnormal rate of lower bone mass in lumbar spine were 57％ (46/81) postmenopausal group,8/15 in POF group,32％ (9/28) in menopausal transition group,12/19 in premature ovarian failure transition group,and osteoporosis was 9％ (7/81),3/15,1％ (3/28)and 0 respectively,their differences were statistically different (P ＜ 0.05) ; the abnormal rate of BMD of left hip and lumbar spine of 11/15 and 12/16 in POF group was higher than 65％ (53/81) in postmenopausal group.In the mean time,the abnormal rate of BMD of left hip and lumbar spine were,12/19 and 10/20 in premature ovarian failure transition group,which were significantly higher than 43％ (12/28) and 39％ (12/31) in the menopausal transition group.Conclusions The menopausal symptoms resulting from hypoestrogenism in natural postmenopausal women are mostly remarkable.The decrease of BMD in lumbar spine is more significant than that of left hip among postmenopausal women.Women with earlier menopause was prone to cause the changes of blood fat and abnormal of BMD,especially HDL-C decreased significantly compared with those natural postmenopause,it is more likely to cause CVD and osteoporosis.

This study is to investigate the effects on the expression of iNOS and production of NO in the osteogenic differentiation process of rat bone marrow stromal cells (rBMSCs) by icariside II. rBMSCs were cultured by adherence screening method. When the culture dishes were covered with 80% cells, the osteogenic induced cultures were adopted. Icariside II was supplemented into the culture at 1 x 10(-5) mol x L(-1). The activity of iNOS, content of NO and osteogenic differentiation markers including alkaline phosphatase (ALP) activity, CFU-FALP and mineralized bone nodules were compared among the icariside II-supplemented group, L-NMAE group, icariside II + L-NAME group and the control. Total RNA was isolated and the gene expression of iNOS, Osterix and Runx-2 was investigated by real-time PCR. Total protein was also isolated and the secretion of iNOS and collagen I was examined by Western blotting. Icariside II can significantly improved ALP activity, CFU-FALP amount and mineralized nodules. Besides, the mRNA level of factors related to the osteogenic differentiation includes Osterix and Runx-2 also enhanced. The secretion of collagen I also promoted significantly. But all of these effects can be inhibited by L-NAME which can specifically inhibit the activity of iNOS. Icariside II enhances the osteogenic differentiation of rBMSCs significantly, but if the activity of iNOS was blocked by L-NAME, the osteogenic differentiation markers decrease accompanied with iNOS and NO decrease, suggesting that icariside II stimulates the osteogenic differentiation via enhancing the activity of iNOS and promoting the generation of NO.

Objective To study the factors influenceing patients' family members to make own relative fac-tors patients' families making decision on refusal of cardiopulmonary resuscitation (CPR) to the critical patients. Method Data were registered based on Utstein Style of 522 patients aged over 15 years, who subjected to in-hos-pital cardiac arrest(CA) in Department of Emergency of The First Affiliated Hospital of Wenzhou Medical College from January 2005 to December 2008. A total of 157 patients' family made refusal decision among the 522 pa-tients, who belonged to the refusal group, and others belonged to the attempt resuscitation group. The associated factors included age, sex, marriage, household register, cause of CA, underlying diseases, capability of activity before CA, life supported with mechanical ventilation, and administration of pressor agents. The refusal decisions were evaluated by using univariate Logistical regression analysis, and then the statistical significant variables were analyzed by using muhivanate Logistical regression analysis. Results Age, household register, cause of CA(car-diac or traumatic),stroke, sudden death, cancer, capability of activity before CA, life supported with mechanical ventilation,and administration of pressor agents were the important factors of making refusal decision (P < 0.01), but sexes or marriage was insignificant related to the refusal decision (P > 0. O5). The independent risk factors re-lated to refusal decision were age (P = 0.034),cancer (P = 0.006),stroke (P = 0.003), and life supported with mechanical ventilation (P = 0.000) in multivariate Logistical regression analysis, but the protective factors were sudden death (P =0.000),cardiac CA (P =0.020) and traumatic CA(P =0.000). Conclusions Age over 60 years, cancer, stroke, and life suppoted with assisted ventilation before CA were factors associated with re-fusal decision making, yet sudden death, cardiac CA and traumatic CA were factors of accepting CPR.

Objective To investigate the changes of tissue factor(TF)and tissue factor pathway inhibitor(TFPI)at different time points after in-hospital eardiopulmonary resuscitation(CPR).and to explore the role of TF and TFPI in CPR.Method From September 2005 to September 2007,24 patients,who suffered from cardiac arrest,were selected from the of emergency medicine department,The First Affiliated Hospital,Wenzhou Medical College.The selected patients were older than 16 years old and had identified cardial :arrest time.All patients were randomly classified into two groups:those who had retum of spontaneous circulation(ROSC)(n:12)and those mthom ROSC(n=12).Ten normal healthy volunteers served as control subjects(n=10).Etiology of the car.diac arrest and clinical characteristics during eardiopulmomry resuscitation were emestered.Serial levels of"IF and TFPI at different time points of 30 min,60 min,6 h,24 h,48 h after CPR were measured by enzyme linked immunosorbent assay(ELISA)after eardarrest and CPR.Data Were expressed as mean±standard deviation twotailed t test and ANOVA and four flod table chi-square test were used for eomparisoll in SPSS 11.5 software,and chaangs were considered as statistically significant if P value was less than 0.05.Results Inpatients with ROSC.TF obviously increased at 30 min after CPR.reached peak at 6 h.1F levels in patients without ROSC wer higher compared to those of the control group and ROSC at the same time point.The levels of TPFI had no significantly differentce at 50 min after CPR,and TFPI,obviously elevated at 60 min after CPR in ROSC group compared to those ofthe control group and without-ROSC group.In comparison with control group,the ratio of TF/FPI at 30 min after CPR in without-ROSC group and ROSC group were marked elevated.The ratio of in without-ROSC group showed marked devations eompare,t to that of ROSC group.In R()sC group.the ratio of THFI peaked at 6 h after CPR and descended a after CPR.Conclusions'11le 1F and TFPI level8 after CPR in patients with in-hospital cardiac obviously increase.The levels of TF and TF/TFPI at 30 min after CPR can be used for predicting the prognosis of patients with in-hospital cardiac arrest.