Objective: To clone,identify and express the Shigalike toxin 2B(Stx2B) subunit gene of EHEC O157∶H7.Methods: A pair of primers were designed based on the Stx2B subunit gene sequence of EHEC O157∶H7.The Stx2B gene was amplified from the EHEC O157∶H7 chromosome by PCR and cloned into the pMD18-T vector.Thereafter,the gene was cut from the pMD18-T vector and cloned into the prokaryotic expression plasmid pET-28a vector.Then the recombinant plasmids were transformed into E. coli BL21(DE3) and the transformed host strain induced by IPTG.The expression protein was detected by SDS-PAGE and Western-blot analyses.Results: The Stx2B gene was successfully cloned into pMD18-T and pET-28a vectors,and the expressed protein identified by SDS-PAGE and Western blot.The molecular mass(Mr) of the expressed product was about 7 500,and the expression rate about 40%. Conclusion: The Stx2B gene was successfully cloned and effectively expressed in the prokaryotic expression system.

Objective:To study the expression of Ki67 and P53 proteins in gingival tissue after wearing four kinds of temporary crowns.Methods:The experiment animal model was established by temporary prosthesis on dogs' teeth using chemical-curing resin,thermal-curing resin,DMG-TEMP crown material and SWIFT composite resin respectively.The immunohistochemistry Envision method was used to measure the expression of Ki67 and P53 proteins in gingival tissue before tooth preparation,after tooth preparation,1-and 2-week,1 month after wearing the crowns.The teeth without any treatment were surved as control.HE stained gingival tissue slices were observed under microscope.Results:The expression of Ki67 and P53 proteins in gingival tissue increased 1 week after wearing the chemical-curing resin crowns or thermal-curing resin crowns(P0.05).No pathological change was observed in all the samples.Conclusion:Chemical-curing resin crowns and thermal-curing resin crowns may increase the expression of Ki67 and P53 proteins in gingival tissue,but DMG-TEMP crowns and SWIFT resin crowns do not.None of the four kinds of temporary crowns may cause abnormal proliferation of gingival epithelia.

Objective:To investigate the role of transforming growth factor β1 (TGFβ1) in the development of Helicobacter pylori ( H. pylori)-associated non-metaplastic atrophic gastritis. Methods:The expressions of TGFβ1, CD68 and smooth muscle actin(SMA) were detected immunohistochemically in 10 patients with mild non-atrophic gastritis, 30 patients with mild non-metaplastic atrophic gastritis, and 32 patients with severe non-metaplastic atrophic gastritis having H. pylori infecion. Meanwhile, three cases of mild non-atrophic gastritis and 4 cases of severe non-metaplastic atrophic gastritis were observed with electron microscope. Results: The count of TGFβ1 positive cells per high-power field (HPF) in severenon-metaplastic atrophic gastritis group (53±22 ) was significantly higher than that in mild non-atrophic gastritis group(22±/HPF) and mild non-atrophic gastritis group(0-3/HPF, P＜0.01). The count of CD68 positive cells in severe non-metaplastic atrophic gastritis group (23±7/HPF) was significantly higher than that in mild non-atrophic gastritis group (13±/HPF) and mild non-atrophic gastritis group (0-3/HPF, P＜0. 01). Correlation analysis showed that the expressions of TGFβ1 and CD68 had a moderate correlation in each group ( r = 0. 634, P＜ 0. 01; r = 0. 699, P＜ 0. 01). Compared with mild non-atrophic gastritis, SMA-positive myofibroblasts and smooth muscle cells in the lamina propria increased in mild and severe non-metaplastic atrophic gastritis. Ultrastructurally, the proliferation of fibroblasts in gastric lamina propria was observed in mild non-atrophic gastritis, while the proliferation of fibroblasts and presence of myofibroblasts could be observed in mild non-metaplastic atrophic gastritis, and there was a parallel phenomenon between myofibroblasts and fibroblasts, as well as smooth muscle cells.Conclusion: Our findings indicate that TGFβ1 expression increases with severity of H. pylori-associated non-metaplastic atrophic gastritis, suggesting that TGFβ1 might play an important role in the development of non-metaplastic atrophic gastritis.

Two cases of Paneth's cell adenoma of the stomach are reported and their morphological, histochemical, immunohistochemical, ultrastructural and immunocytochemical features are described. Both surgical specimens showed varied dysplasia with severe atrophic gastritis and intestinal metaplasia. 60%-70% of the tumor cells contained coarse eosinophilic cytoplasmic granules that are PAS positive but unstained by alcian blue at pH 2.5, and exhibited positive antiserum immunoreaction for ly-sozyme. The Paneth's nature of these granules was confirmed by electron microscopy and immunoelec-tron microscopy. In addition, some endocrine cells were detected in the neoplastic glands. The characteristics of Paneth's cell adenoma of the stomach are discussed and a close relationship between the histogenesis and intestinal metaplasia is suggested.

AIM:To discuss the technique for preparing high-purity solanesol from tobacco. METHODS:The pre-treated tobacco leaves were extracted with mixed solvent of petroleum ether and ethanol,and saponified to obtain solanesol extract; The solanesol extract got through redissolution and dewaxing,then crystallized in the temperatare of -18 ℃; Finally purified by macroporous resin(YPR-Ⅱ,D-1300,HZ-816,HZ-802). RESULTS:The ratio of mixed solvent of petroleum ether and ethanol was 1 ∶ 4,ratio of liquid to materials was 1 ∶ 12,solanesol purity was about 32% after saponification,then crystallization made the purity up to 63. 4% ,finally purified through macroporous resin,the solanesol purity reached above 93. 6%. CONCLUSION:Through the general solvent extraction,saponification,crystallization and macroporous resin adsorption process,the solanesol purity of the product reaches higher than commonly expected values.

ObjectiveTo evaluate the efficacy and safety of tirofiban in treatment of ST segment elevation acute myocardial infarction(STEAMI) no-reflow and acute thrombosis after emergency percutaneous coronary intervention(PCI). MethodsForty patients which were made definite diagnosis of STEAMI were intra-coronary artary injection fortirofiban after emergency PCI stenting occured no-reflow and acute thrombosis. First,the dose of 0.4μg·kg-1·min-1 was given from intra--coronary artary injection of tirofiban within three minutes, after 30min the dose were given 0.1μg·kg-1·min-1 for 48 hours. ResultsThe no re-flow and acute thrombosis was completely disappeared within five minutes,at the time,side effect with in one week was not observed. ConclusionsTirofiban treatment by direct injection in coronary arteries combined with emergency PCI, can increase the repeffusion rate of infarction related vessel in AMI patients,and improve TIMI reflow. This reperfusion method was effective and safe.

Objective To compare the clinical outcomes of Chinese rapamycin-eluting stents (Firebird stents) and imported paclitaxel-eluting stents ( Taxus stents ) in the treatment of acute myocardial infarction. Methods Ninety-seven patients with ST segment elevated acute myocardial infarction were treated with Firebird stents (in 51 patients) and Taxus stents (in 46 patients). The death rate, re-acute myocardial infarction, target lesion revascularization (TLR) ,and major adverse cardiac event (MACE) within 9 months after percutaneous coronary intervention ( PCI) were observed between the two groups. Results The rate of successful stent-implantation, angina,death, re-acute myocardial infarction, TLR and MACE was 100% ,9. 8% ,0% ,2. 0% ,0% , 11. 8% in the Firebird stent group and 100% ,8. 7% ,0% ,2. 2% ,0% ,0% and 10.9% in the Taxus stents group within 9 months after PCI. There was no significant difference between the two groups. Conclusions There is no significant difference in the clinical effect between the Firebird stent group and Taxus stent group within 9 months after PCI. However, the effect-cost ratio is better in the Firebird stent than the Taxus stent.

ObjectiveTo investigate the association of hypertensive target organ damage with abnormal ankle brachial index (ABI) in high-risk hypertensive patients.MethodsDuring December 2008 to May 2009,a cross-sectional study was conducted to investigated the prevalence of abnormal ABI (ABI ＜ 0.90) in 2674 community-dwelling,hypertensive patients,who aged ＞ 40 years,without coronary heart disease,stroke/transient ischemic attack or known arteriosclerosis,from 18 centers in China. Data were acquired through history,physical examination,laboratory and other diagnostic tests.ResultsThere were 2615 subjects eligible for the full analysis set. The high-risk hypertensive patients with arterial wall thickening,arterial wall thickening and slightly elevated serum creatinine had a higher prevalence of abnormal ABI than their counterparts respectively ( P ＜ 0.05 ). Compared with the normal group,the abnormal ABI group had a higher serum creatinine level on average (P ＜ 0.01 ).After adjustment for certain factors including investigation center,demographic factors,cardiovascular disease (CVD) risk and CVD risk factors using an unconditional logistic regression model,arterial wall thickening ( OR 2.416,95％ CI 1.395-4.183,P =0.0016 ) and slightly elevated serum creatinine ( OR 3.377,95％ CI 1.267-8.997,P =0.0149) were positively associated with abnormal ABI. However,arterial wall thickening (OR 0.988,95％ CI 0.576-1.695,P=0.9664) and microalbuminuria (OR 1.389,95％ CI0.685-2.817,P=0.3621)were irrelevant to abnormal ABI.Conclusions So far as a high-risk hypertensive patient is concerned,there are significant statistical correlations between arterial wall thickening and/or slightly elevated serum creatinine and an abnormal ABI,but no significant statistical correlations between arterial wall thickening or microalbuminuria and an abnormal ABI is observed.

Summary: Receptor mediated gene delivery is a new gene transfer strategy. Asialoglycoprotein receptor (ASGP-R), the receptor of asialoorosomucoid (Asor), is specially expressed on the surface of hepatocyte. In this paper, the nuclide 131I was combined with Asor to form a kind of soluble nuclide-protein complex, which can be specifically endocytosed into hepatocyte by ASGP-R. After intravenous injection of the complex into experimental animals, the deposition of Asor in vivo and the targeting quality of hepatocyte was detected by ECT. This research testified the feasibility of targeting Asor complex delivery to hepatocyte mediated by ASGP-R in vivo, and provided foundation for the genetic diagnosis and gene therapy of hepatic cell-related diseases.

Objective To evaluate the effect of pitavastatin on blood glucose in patients with hypercholesterolemia,and to investigate the efficacy of pitavastatin in diabetic patients combined with hypercholesterolemia.Method This study was a 12-week,multi-center,open-label,without parallel-group comparison,phase Ⅳ clinical trail.Results Contrasting to baseline,the prevalences at week 4 and 12 post-treatment of abnormal fasting plasma glucose (FPG) and glycosylated hemoglobin Alc (HbA1c)( FPG:14.2％ vs 14.1％ and 11.0％ ; HbA1c:14.3％ vs 15.1％ and 16.1％ ) in the safety set subjects without diabetes mellitus (DM),as well as in those with DM but not taking glucose-lowering drugs (FPG:7/7 vs 4/7 and 5/7; HbAlc:5/5 vs 4/4 and 5/5) had no significant changes (all P vaules ＞0.05).Contrasting to baseline,the levels of TC [ (6.51±0.94) mmol/L vs (5.12 ±0.93) mmol/L and (4.54 ±1.00) mmol/L],LDL-C [(4.11 ±0.79)mmol/L vs (3.02 ±0.81) mmol/L and (2.51 ±0.70)mmol/L] and TG [2.10(1.53,2.54) mmol/L vs 1.62(1.26,2.00) mmol/L and 1.35(1.10,1.86)mmol/L]at week 4 and 12 post-treatment in the per protocol set 55 subjects with DM were significantly reduced (all P values ＜ 0.05 ) ; 33.3％ of subjects at high risk and 10.0％ of subjects at very high risk had achieved a TC target value; 55.6％ of subjects at high risk and 40.0％ of subjects at very high risk had achieved a LDL-C target value.Conclusion Pitavastatin has a safe effect on blood glucose and it could be used to treat diabetic patients combined with hypercholesterolemia in China.