Objective:To clone human canstatin cDNA,express it as fusion protein in E.coli,purify the fusion protein and determine its biological activity.Methods:Total RNA was extracted from Chinese placenta tissue.The canstatin cDNA was amplified by RT-PCR,then cloned into pTYB1 and sequenced;Prokaryotic expression vector pTYB1-hCAN was constructed;fusion protein was expressed by using IPTG induction and purified by chitin column.The biological activity of the purified fusion protein was identified by using the chick embryo chorioallantoic membrane assay(CAM).Results:RT-PCR product was about 700 bp,its sequence was the same as that of canstatin reported.Prokaryotic expression vector pTYB1-hCAN of human canstatin was constructed successfully,and the fusion canstatin protein was expressed in E.coli.The purity of the protein was 91.6%.The fusion protein could suppress the angiogenesis in CAM.Conclusion:The active human Canstatin is cloned and expressed as a bioactive form.

AIM:To investigate the effects of cancerous inhibitor of protein phosphatase 2A (CIP2A) over-ex-pression on the proliferation and apoptosis of gastric epithelial cells .METHODS:The mRNA expression of CIP2A and cy-clin D1 in the tissues of normal gastric mucosa and gastric polyps was detected by RT -qPCR.The GES-1 cells were divided into control group, Ad-emp group and Ad-CIP2A group.The cell proliferation ability was detected by MTT assay and BrdU assay , and the cell apoptosis was analyzed by flow cytometry .The expression of apoptosis-related molecules was determined by Western blot and RT-qPCR.The levels of inflammatory cytokines were measured by ELISA after GES-1 cells were infec-ted with Ad-emp and Ad-CIP2A.Furthermore, the protein levels of p-Rb, E2F1 and cyclin D1 in the GES-1 cells was de-termined by Western blot after transfected with E2F1 siRNA.RESULTS:The expression of CIP2A and cyclin D1 in ade-nomatous gastric polyps tissues was significantly higher than that in normal gastric mucosa tissues , and no significant change of that between hyperplastic gastric polyps tissues and normal gastric mucosa was observed .After transfected with CIP2A, the proliferation ability of GES-1 cells was increased , the cell apoptosis was inhibited , the concentrations of IL-1βand IL-10 was up-regulated and the protein levels of p-Rb, E2F1 and cyclin D1 were increased, while the protein levels of p-Rb, E2F1 and cyclin D1 were significantly decreased after transfected with E2F1 siRNA.CONCLUSION: CIP2A promotes the proliferation and inhibits the apoptosis of GES-1 cells by activating Rb/E2F1.

Object To develop a method for the quantitative determination of salvianolic acid A in rat plasma. Methods HPLC method and Hypersil ODS column (4 6 mm?200 mm, 10 ?m) were used, the mobile phase was acetontrile water (1∶2 65, adjusted pH to 2 5 with formic acid), at a flow rate of 0 9 mL/min, the UV detection wavelength was 285 nm. Cinnamic acid was used as internal standard. Results The recovery of the method was in the range of 98 9%～106 9%, the recovery of the extraction was over 83%. The within day precision and the day to day precision were both lower than 8 4%. Conclusion The method is appropriate for the determination of salvianolic acid A when pharmacokinetics of salvianolic acid A is studied in rat plasma.

Objective To investigate the function of serum soluble intercellular adhesion molecule-1(sICAM-1) in acute pancreatitis(AP).To study the significance of continuous monitoring sICAM-1 in differential diagnosis of AP.Methods A total of 60 patients were investigated;36 patients with acute edematous pancreatitis(AEP),and 24 with acute haemorragic necrotic pancreatitis(AHNP).The concentrations of sICAM-1 in the patients' serum were measured serially over a period of 7d by enzyme-linked immunosorbent assay(ELISA).Results There was significant difference between AEP group and AHNP group(P

Objective:To evaluate the effect of of twin block appliance in the early treatment of class Ⅱ malocclusion.Methods:20 cases with class Ⅱ malocclusion were treated with Twin block appliance.Cephalometric analysis was used to evaluate the effects.Results:The twin block appliance could stimulate the growth of mandible and lead to retrocline of the upper incisors and procline of the lower incisors.Conclusion:The twin block appliance can achieve remarkable effects of growth modification of dentognathic system.

Objective To make the clinical evaluation of Targis/Vectris restorations. Methods 337 cases with Targis/Vectris restorations were rechecked 12～50 months after cementation respectively. The restorations were evaluated clinically according to the USPHS standard. Results The changes of Targis/Vectris restorations in colour match, boundary line, wear and tear, marginal adaptation and surface grain were not obvious within 12～50 months. The secondary caries were not found. The successful rate of the restorations was 95.4 %.Conclusion The clinical prosthetic results of Targis/Vectris restorations are reliable.

Objective To investigate the dynamic changes in brain natriuretic peptide (BNP) and the outcome of patients with chronic congestive heart failure (CHF) after the treatment with metoprolol. Methods Sixty-five patients with CHF were admitted to the Forth Affiliated Hospital of Zhengzhou University during April 2004 to April 2007, and they fulfilled the diagnostic criteria of chronic CHF issued by New York Heart Association (NYHA). Of all the patients the left ventricular end-diastolic dimension (LVEDD) ≥65mm. All the patients were randomly assigned into metoprolol group (34 cases) and control group (31 cases), and received conventional therapies [rest, reduced sodium intake, administration of digitalis, diuretic, angiotensin-converting enzyme inhibitor (ACEI) and aldosterone antagonists, etc.]. The patients in metoprolol group were given metoprolol (6.25mg as initiative dose and dose was increased according to illness). NYHA functional classitication, cardiothoracic ratio on chest X-ray film, cardiac output (CO), cardiac index (CI), left ventricular ejection fraction (EF) and LVEDD in echocardiogram were observed, and the plasma BNP level was determined with an immunoradiometric assay (IRMA). Results After 5 months of treatment, the clinical symptoms of patients were improved obviously and p1asma BNP level was significantly lowered in both groups, while p1asma BNP level was significantly lower in metoprolol group compared with control group (200.6?12.1 vs 120.7?14.1, P

PET-CT plays a increasing important role in the treatment of lung cancer.Compared with CT,the advantage of PET-CT is obvious,which is more accuracy when determine a metastasis and identify inflammatory lesions.What's more,on the basis of the accuracy staging,PET-CT change many strategies which were based on traditional image,resulting in improving the treatment effect.While the assessment and prediction of the efficacy by PET-CT also exerted an important influence on the formulation of treatment strategies.

Objective:To construct a recombinant adenoviral vector carrying the specific siRNA of Survivin gene,and to observe its effect on the expression of Survivin gene and on the radiosensitivity of cancer cells in tumor-bearing mice.Methods:The specific siRNA of Survivin gene was designed and synthesized,and a recombinant adenovirus AdEGFP-siRNA was subsequently constructed.SMMC 7721 xenograft models were established with nude mice and were divided into the following 5 groups:siRNA+radiotherapy and siRNA groups(intratumoral injection of AdEGFP-siRNA),siRNA(-)group(injected every other day with AdEGFP-siRNA[-],2?108 pfu/100 ?l per time,total 5 times),pure radiotherapy group and blank control groups(injected with the same volume of normal saline).On day 10,12,14,and 16,the mice in siRNA+radiotherapy and pure radiotherapy groups were given 5 Gy/time radiotherapy.The tumor volumes were measured regularly.The expression of Survivin in tumor tissues was determined immunohistochemically.Results:Adenovirus AdEGFP-siRNA harboring the specific siRNA of Survivin gene and enhanced green fluorescent protein gene(EGFP)was successfully recombined.The growth of SMMC 7721 xenografts in nude mice was inhibited after injecting AdEGFP-siRNA,with the inhibition rate being 56.2%.The inhibition rate in AdEGFP-siRNA therapy + radiotherapy increased to 82.6%.Immunohistochemistry study showed that the specific siRNA markedly silenced the expression of Survivin gene in hepatocarcinoma cells.Conclusion:The specific siRNA can markedly silence Survivin gene and subsequently inhibit the growth of cancer;meanwhile,it can also increase the radiosensitivity of cancer cells so as to improve the treatment effect.

Acute respiratory distress syndrome (ARDS) is a severe respiratory condition that is characterized by rapidly progressive hypoxemia with noncardiogenic pulmonary edema.Despite the improvement of therapeutic methods,the mortality of ARDS is in the range of 40％-50％ all over the world.Some studies have shown that a significant number of patients with ARDS had acute cor pulmonale (ACP),and ACP is independently associated with the mortality of patients with ARDS,which has attracted wide attention in recent years.This paper reviewed recent related studies,summarized the prevalence,pathogenesis and diagnostic approaches of ACP in ARDS,especially echocardiography which was considered as a cornerstone for ACP diagnosis,and elucidated the beneficial effects of right ventricular protective ventilatory strategy and prone-positioning on the pulmonary vasculature and right heart,in order to provide a novel idea for the therapy of ACP in ARDS.