AIM:To investigate the effect of the N-terminal 24-amino acid (N24) overexpression in p55γre-gulatory subunit of phosphatidylinositiol 3-kinase ( PI3K) on the cell adhesion of human gastric carcinoma cell MGC 803. METHODS:MGC803 cells, which stably expressed GFP-N24 fusion protein and GFP alone , were generated by transfec-tion with pEGFPN-24 plasmid and control plasmid pEGFP-C1, respectively.The morphological change of the cells was ob-served under inverted microscope .The expression of GFP-N24 fusion protein was detected by Western blot .The adhesion of the cells was determined by cell adhesion assay .The effects of GFP-N24 fusion protein on the expression of E-cadherin andβ-catenin were analyzed by Western blot .The expression and secretion of matrix metalloproteinase 9 (MMP9) and u-rokinase-type plasminogen activator ( uPA ) in the MGC803 cells were detected by gelatin zymography .RESULTS:MGC803/GFP-N24 cell line steadily expressed GFP-N24 fusion protein and MGC803/GFP cell line steadily expressing GFP were successfully established , but the expression of fusion protein GFP-N24 was lower than that of the control protein GFP.The morphological changes of the transfected cells from paving stone to fibroblast cell form after gene transfection , and the cytoplasm secretory granules were increased significantly .The cell adhesion to fibronectin and collagen decreased after GFP-N24 transfection .GFP-N24 fusion protein increased the expression of cell adhesion molecule E-cadherin and de-creased the wnt signal pathway molecule β-catenin in the MGC803 cells.However, it did not affect the expression and se-cretion of tumor metastasis-related proteins MMP9 and uPA.CONCLUSION:Overexpression of N24p55γinhibits cell ad-hesion by influencing the expression of E-cadherin and β-catenin in the MGC803 cells.

AIM:To study the protective effects of extract from Tribulus terrestris L on adriamycin-induced damage to myocardial mitochondria in rats. METHODS: Rats were given intraperitoneal injection of adriamycin (ADR) to induce the damage to myocardial mitochondria. After continuous ig of extract from Tribulus terrestris L, the every experimental group was observed myocardial ultrastructure with electron microscope, the mitochondrial membrane potential (MMP) and the activity of glutathione peroxidase (GSH-Px) ill myocardial mitochondria were determined. RESULTS: ADR of 20 mg/kg was given to rats intermittently induced severe damage to myocardial mitochondria in rats, decreased MMP and the activity of GSH-Px. Extract from Tribulus terrestris L middle- and high-dosage groups (30 mg/kg, 45 mg/kg) could reduce cardiotoxicity of ADR at the ultrastructure level, and increase MMP and the activity of GSH-Px. CONCLUSION: Extract from Tribulus terrestris L may decreased the damage to myocardial mitochondria induced by adriamycin, and has a protective effect on adriamycin-induced cardiotoxicity.

Objective To investigate the clinical characteristics of multisystem impairment of Sj(o)gren's syndrome (SS) to make quick and accurate diagnose and treatment to decrease the rate of misdiagnosis and mistreatment.Methods A case of thrombotic thrombocytopenic purpura with SS was reported and the related literatures were reviewed.The clinical manifestations,laboratory examinations,diagnostic criteria and therapy of thrombotic thrombocytopenic purpura with SS were analyzed.Results Purpura haemorrhagica,hemolytic anemia,central nervous systemic symptoms,kidney impairment and fever were the representative clinical manifestations of thrombotic thrombocytopenic purpura with SS.Plasmapheresis was the most effective therapy with the background treatment with glucocorticosteroid and immunosuppressive agents.Conclusion Thrombotic thrombocytopenic purpura with SS presents a complicated clinical manifestations,so misdiagnosis and mistreatment is very frequent.More attention should be paid to this specific situation because the disease is rare and has high mortality rate.

OBJECTIVETo study the effects of different doses of taurine (Tau) on calcium ion concentration ([Ca2+]i) and ATPase on cardiocyte membrane of diastole heart failure rats.

METHODDiastole heart failure model was established by the coarctation of abdominal aorta. Four weeks after operation, forty diastole heart failure rats were divided into four groups randomly as follows, model (normal saline 2 mL), taurine (400 mg x kg(-1) x d(-1)), taurine (200 mg x kg(-1) x d(-1)), taurine (100 mgx kg(-1) x d(-1)), with 10 rats for each group (n=10), and 10 sham operation rats was taken as control(normal saline, 2 mL). After 4 weeks administration, Isolate single cardiocyte by enzymatic isolation method which were loaded with Ca2+-sensitive fluorescent indicator Fluo-3/AM. [Ca2+]i was measured by laser scanning confocal microscope [LSCM], and represented it by fluorescent intensity [FI]; ATPase activity of cell membrane was measured by the method of enzymatic reaction chromatometry.

RESULTCompared with the control group, [Ca2+]i in cardiocyte increased markedly and the ATPase activity of cardiocyte membrane decreased significantly in the model group. Compared with the model group, fluorescent value decreased and ATPase activity increased significantly in Tau high-dose group; fluorescence value and ATPase activity decreased significantly in Tau mid-dose group; fluorescent value decreased and ATPase activity increased significantly in Tau low-dose group.

CONCLUSIONLarge dosage of Tau can increase ATPase activity on cardiocyte membrane, improve [Ca2+]i in cardiocyte and antagonise calcium overload of DHF rats.

Adenosine Triphosphatases ; metabolism ; Animals ; Calcium ; metabolism ; Diastole ; drug effects ; Disease Models, Animal ; Female ; Heart Failure ; drug therapy ; metabolism ; Male ; Myocardium ; pathology ; ultrastructure ; Myocytes, Cardiac ; drug effects ; metabolism ; pathology ; Rats ; Rats, Wistar ; Taurine ; administration & dosage ; pharmacology ; therapeutic use

Objective To explore the potential mechanisms of non-small cell lung carcinoma cells to rsTRAIL protein-induced apoptosis by phosphatidylinositol 3′-kinase (PI3K/Akt)inhibitor LY294002,and to provide new ways to increase killing activities of rsTRAIL protein for non-small cell lung cancer.Methods The A549 cells at logarithmic growth phase were selected and randomly divided into rsTRAIL group and LY294002+rsTRAIL group. The inhibitory rate of growth of the A549 cells was tested by MTT assay.The cell cycle and apoptotic rate were detected by flow cytometry analysis. The expression levels of Ser473 phosphorylated form of Akt (p-Akt),c-FLIPL protein and Bcl-2 protein in the A549 cells in two groups were analyzed by Western blotting method. Results The inhibitory rate of growth of the A549 cells in LY294002+rsTRAIL group (74.6 %± 2.63%)was higher than that in rsTRAIL group (5.61% ± 0.32%) (P< 0.05 ). Compared with rsTRAIL group, the percentage of the cells at G0/G1 phase in LY294002+rsTRAIL group was increased(P<0.05)and the percentage of the cells at S phase was decreased(P<0.05).The apoptotic rate of the A549 cells in LY294002+rsTRAIL group (61.5%±3.02%)was higher than that in rsTRAIL group (3.21%±0.96%)(P<0.05). The Western blotting results showed that the expression levels of p-Akt, c-FLIPL and Bcl-2 proteins in the A549 cells in LY294002+rsTRAIL group were decreased (P<0.05 )and the ratio of Bax/Bcl-2 was increased (P<0.05 ) compared with rsTRAIL group.Conclusion LY294002 can increase the killing activity of rsTRAIL protein in A549 cells by inhibiting the activity of PI3K.

Objective To evaluate the life quality in patients who underwent laparoscopic appendectomy resection and open appendectomy resection for choledocholithiasis.Methods A survey was made on 30 patients who underwent laparoscopic appendectomy resection and 30 patients who underwent open appendectomy resection.The quality of life was measured with the Gastrointestinal quality of life index (GIQLI) preoperation and 2,5,10,16 weeks after the operation.Results There were no significant difference in the GIQLI scores.The GIQLI scores were 121,115,126,131,and 133 respectively in preoperation and 2,5,10,and 16 weeks after laparoscopic appendectomy resection.The scores of GIQLI were 122,109,120,127,and 129 respectively in preoperation and 2,5,10,and 16 weeks after open appendectomy resection.Conclusion The GIQLI scores from laparoscopic appendectomy resection group is better than that from the open appendectomy resection group.

Objective:To investigate mechanism of TRAIL-resistance in A549 cells ( a cell line of non-small cell lung carcino-ma cells) due to Akt phosphorylation .Methods:A549 cells were treated with Akt inhibitor Perifosine and rsTRAIL protein individual-ly and in combination.The expressions of Akt phosphorylation(p-Akt),c-FLIPLL and caspase-8 were detected by Western blot.The apoptotic rate of the A549 cells treated was detected by flow cytometry and the cell proliferation was evaluated by MTT assay .Results:A549 cells showed the increased level of Akt phosphorylation mediated by rsTRAIL protein .Treatment with the Akt inhibitor Perifosine induced a suppression of Akt activation in A 549 cells and a concomitant decrease in the expression of c-FLIPLL .As a result, Perifosine significantly enhanced TRAIL-induced apoptosis rate of (76.5 ±3.02)%and cytotoxic rate of (83.2 ±2.54)%by promoting the activ-ity of caspase-8.Conclusion:Akt activity promotes A549cells survival against TRAIL-induced apoptosis and that the cytotoxic effect of rsTRAIL protein can be enhanced by modulating the Akt phosphorylation in human non -small cell lung carcinoma cells .

Objective:To determine the prevalence of Helicobacter pylori (Hp) infection in orthotopic liver transplantation (LT) recipients, and to study the efficacy and safety of Hp eradication therapy.Methods:13C-urea breath test was carried out between July 2018 and October 2019. There were 104 males and 26 females with an average age of 52.1 year for these LT recipients who were regularly followed-up in the Organ Transplant Center, the Affiliated Hospital of Qingdao University. Propensity scoring was used to match age and gender in a ratio of 1∶3. A healthy group of individuals consisting of 299 males and 91 females, with an average age of 51.8 years, were selected as the control group also in a ratio of 1∶3. All patients underwent 13C-urea breath test to evaluate Hp infection and results of quadruple therapy. Results:The prevalence of Hp infection among the LT recipients was 18.5%(24/130) which was significantly lower than the control group 36.4% (142/390) (χ 2=14.400, P<0.001). Among the recipients who received LT and 13C-urea breath test for less than 1 year, 1-3 years and more than 3 years, the prevalences of Hp infection were 14.3% (6/42), 17.8%(8/45) and 23.3%(10/43), respectively. Although the prevalence of Hp showed a gradually increasing trend, no correlation between Hp infection and duration from LT was observed (χ 2=1.321, P=0.517). Seventeen Hp positive recipients underwent Hp eradication therapy. The success rate of Hp eradication was 100.0%(17/17). Immunosuppressant concentration was monitored regularly in 10 patients. During Hp eradication, the blood concentration of immunosuppressant increased from 1.7 to 3.6 times, and the drug dosage was reduced to one half to one third of what it was before Hp eradication. Seven of these 17 recipients suffered from adverse effects caused by increased levels of blood concentration of immunosuppressants, including headache, hypertension, insomnia and irritability. Normal liver and kidney functions were observed during eradication. Conclusion:In this study, the prevalence of Hp infection among liver transplant recipients was 18.5%. The success rate of Hp eradication was 100% using bismuth-containing quadruple therapy. The dosage of immunosuppressant required to be monitored, and if necessary, adjusted during eradication to reduce adverse side effects.

Objective To explore the diagnostic value of k ras mutation detection of specimen obtained by endoscopic ultrasound-guided fine needle aspiration (EUS-FNA)for pancreatic cancer.Methods Seventy-eight patients with pancreatic carcinoma and 49 patients with pancreatic benign diseases were collected from January 2013 to December 2015.All the patients underwent EUS-FNA and cell or tissue samples were collected.DNA was extracted from the samples,and the codon 12 and 13 mutation of K-ras gene was detected by specific nano capture probe.Liquid cytology was also conducted.The sensitivity and specificity of the two methods were compared.Results The K-ras mutation rate was 92.3％ (72/78) in 78 patients with pancreatic carcinoma,which was obviously higher than the mutation rate of 20.4％ (10/49) in 49 patients with pancreatic benign tumors,and the difference was statistically significant (x2 =68.002,P =0.000).The sensitivity of the cytology examination of EUS-FNA specimens in the diagnosis of pancreatic carcinoma was 75.6％,the specificity was 87.8％,Youden index was 0.634,the positive and negative predictive value was 6.196 and 0.227.The detection of K-ras mutations had a sensitivity of 92.3％ and a specificity of 79.6％ in the diagnosis of pancreatic carcinoma,the Youden index was 0.719,the positive and negative predictive value was 4.524 and 0.096.K-ras mutation detection had a higher sensitivity compared with cytology,and the difference was statistically significant (x2 =8.47,P =0.004).The sensitivity was 94.9％ and specificity of 95.9％ using the combination of K-ras mutations and cytology for diagnosing pancreatic cancer,and the specificity was obviously increased compared with only k ras mutation detection and the difference was statistically significant (x2 =6.13,P =0.013).Conclusions K-ras mutation detection of EUS-FNA specimen using nano capture probe system can significantly improve the sensitivity of diagnosing pancreatic cancer,and the sensitivity and specificity could be further improved when combined with cytology.