Objective To study a rapid and sensitive method for determination of virulence-associated genes in O1El Tor,O139,non-O1/non-O139Vibrio cholerae strains.Methods Five pairs of primers were designed respectively ac-cording to cholera toxin sub-unit A gene(ctxA),accessory cholera enterotoxin gene(ace),zonula occludens toxin gene(zot),toxin coregulated pilus A gene(tcpA)and toxR regulatory protein gene(toxR).Multiplex PCR(MPCR)procedures for simultaneously detecting those five genes were established.The gene information of the virulence-associated genes in the Vibrio cholerae strains was obtained through agar gel electrophoresis for products of single amplification of the MPCR.Results The five virulence-associated genes in the positive control Vibrio cholerae O139(MO45strain)could be detected and the results were correct,which could meet the designed request for the method.In the other tested strains(O1EL Tor,O139,non-O1/non-O139)could be detected1to5kinds of the virulence-associated genes.Based on the results of the variety of carried virulence-associated genes,the tested Vibrio cholerae strains could be classified as5kinds of genetypes,and the Vibrio cholerae could be distinguished between toxic and non-toxic strains.The sensitivity of the MPCR approach reached to10 2 cfu/ml.Conclusion This method is rapid,specific and sensitive,which possess great value for practical application.

OBJECTIVETo analyze the risk factors of liver metastasis in patients after radical resection of pancreatic cancer.

METHODSOne hundred and twenty-four patients with non-metastatic, resectable pancreatic cancer treated in our department between 2006 and 2012 were included in this study. All of these patients underwent resection of the primary tumor combined with extensive lymph node dissection. The development of postoperative liver metastases was carefully followed up, and the clinicopathological factors and molecular characteristics were evaluated by univariate analysis and multivariate logistic regression using SPSS 16.0 software.

RESULTSForty-eight cases of liver metastases were found among the 124 cases of pancreatic cancer after radical surgery (38.7%). The rate of liver metastasis of pancreatic cancer after radical surgery in the age groups < 40, 40-60, and > 60 were 68.8%, 33.3% and 35.1%, respectively. The rate of liver metastasis in the body mass index (BMI) group < 20 kg/m2, 20-25 kg/m2, and > 25 kg/m2 were 21.6%, 44.1% and 52.6%, and the rate of liver metastasis in the time between the onset and diagnosis groups ≥ 3 months and < 3 months were 59.4% and 31.5%, respectively. The rate of liver metastasis in patients with preoperative fatty liver was 14.3% and it was 43.7% in patients without preoperative fatty liver. The rate of liver metastasis in patients of histological high, medium and low grade was 10.0%, 35.4% and 49.0%, respectively. The rate of liver metastasis in patients with venous tumor thrombus was 68.8% and it was 34.3% in patients without venous tumor embolus. The rate of liver metastasis in patients with postoperative chemotherapy was 31.2% and it was 51.1% in patients without postoperative chemotherapy. All those differences had statistical significance (P < 0.05). Univariate analysis revealed that age, body mass index (BMI), time between the onset and diagnosis, preoperative fatty liver, histological grading, tumor invasion depth, venous tumor embolus, and postoperative chemotherapy were significantly related to postoperative liver metastasis. Multivariate analysis revealed five statistically independent risk factors for postoperative liver metastasis: BMI, time between onset and diagnosis, preoperative fatty liver, histological grading, and venous tumor embolus.

CONCLUSIONSOur data suggest that patient's BMI, time between onset and diagnosis, histological grade, and venous tumor embolus are significantly correlated with postoperative liver metastases in patients with pancreatic cancer. Pancreatic cancer patients with preoperative fatty liver have less postoperative liver metastasis.

Adult ; Aged ; Body Mass Index ; Humans ; Liver Neoplasms ; secondary ; Lymph Node Excision ; Middle Aged ; Pancreatic Neoplasms ; pathology ; surgery ; Regression Analysis ; Risk Factors

Objective To analyse tratment strategies and to evaluate the relation between different therapies and survival rate of patients of with asynchronous liver metastases after pancreatic cancer surgery (PCLM).Methods From January 2006 to January 2012,48 patients with PCLM were included in this study,and their medical records were retrospectively analyzed.Results Among the 48 patients,27 cases of liver metastases were found within six months after surgery,and the survival rate for 1,3 and 5 years was 22.2％,3.7％ and 0％,respectively,with the median survival of 6 months,and 21 cases of liver metastases were found after six months,and the survival rate for 1,3 and 5 years was 85.7％,30.6％ and 9.2％,with the median survival of 15 months,and the difference between the two groups was statistically significant (P ＜ 0.01).After pancreatic cancer surgery and adjuvant gemcitabine chemotherapy,the probability of liver metastases was 33.3％ (8/24) within six months,the median disease-free survival time was 8 months and the disease-free survival rate for 1,3 and 5 years was 20.8％,4.3％ and 0％.For patients without adjuvant gemcitabine chemotherapy,the probability of liver metastases was 79.2％ (19/24),the median disease-free survival time was 3 months and the disease-free survival rate for 1,3 and 5 years was 4.2％,0％ and 0％,and the difference between the two groups was statistically significant (P ＜ 0.01).The overall survival for patients undergoing resection of liver metastases combined with gemcitabine treatment was better than the other groups (P ＜ 0.01).And the overall survival for patients undergoing transhepatic arterial embolization (TACE) combined with gemcitabine treatment was better than TACE group,gemcitabine group or the observation group (P ＜0.05).There were no difference in overall survival between TACE group,gemcitabine group and observation group.Conclusions Pancreatic cancer patients who develop liver metastasis within six months after surgery have poor prognosis,but postoperative chemotherapy can delay the development of liver metastasis.For patients with resectable lesion,resection of asynchronous liver metastasis is the treatment of choice,and TACE combined with gemcitabine has better efficacy than that of single treatment.

Objective To discuss the individualizing schedule of endovascular treatment for intracranial venous sinus thrombosis based on the clinical feature and the stage of the disease.Methods Forty-three patients diagnosed with intracranial venous sinus thrombosis in Xuanwu Hospital,Capital University of Medical Sciences and in Zhongshan Hospital Xiamen University during period of August 2010 to August 2011 were treated with endovascular therapy designed individually based on the clinical stage of the disease development.Of all 43 cases,22 cases with acute onset (＜ 1 week after the onset) were treated with standard anticoagulant therapy ; 11 cases who failed to respond to anticoagulant therapy and 8 cases with subacute onset (1 week to 1 month) received intravenous thrombolysis and mechanical thrombus maceration;and 13 cases with chronic course (＞ 1 month) were given mechanical thrombus maceration combined with balloon dilation vascular surgery or stent-assisted venous sinus surgery,with taking anticoagulant for 12months.The patients were followed up for 3 to 6 months (mean 4.5 months).Results Their symptoms and signs were all significantly improved,with headache relief in 29 cases(67.4％),vision improvement in 28cases (28/31,90.3％),cerebrospinal fluidpressure decrease to normal level in 32 cases(32/43,74.4％) ;no improvement in 1 case,and a complication of subdural hematoma in 1 case.Three months follow-up of 39 patients,symptoms disappeared,the pressure cerebrospinal fluid in 36 cases returned to normal (94.3％),the papilledema in 37 cases subsided (94.9％).After 6 months,16 patients were followed up with an angiography; 8 of them were found venous sinus clear,5 were found partial recanalization of venous sinus trunk,cortical veins and deep venous was partially compensation,and 3 cases with stent-assisted venous sinus surgery were found the stent not shifted or collapsed,and venous sinuses maintained patency.Conclusions Endovascular treatment for patients with intracranial venous sinus thrombosis should be treated with individually designed therapy based on their clinical features and the stage of the disease.The individualized treatment was effective and safe.

Objective To study the effect and safety of intra-arterial thrombolysis combined with mechanical ad-juvant in the treatment of acute large artery occlusive infarction of anterior circulation and posterior circulation. Methods Fourty-tow patients were divided into anterior circulation group (24 cases) and posterior circulation group(18 cases). The recanalization rate, NIHSS score (National Institute of Health stroke scale), GCS score ( Glasgow coma score,GCS), BI ( Barthel Index) excellent rate, symptomatic intracranial hemorrhage rate and mortality was analyzed after intra-arterial thrombolysis combined with mechanical aids therapy. Results The anterior circulation group was mainly caused by car-diogenic embolism (15/24, 62.5%) and posterior circulation group was mainly caused by atherosclerosis thrombosis (5/18, 72.2%). The NIHSS score was significantly lower after treatment (8.3±4.9 vs. 8.1±5.7) than before treatment(15.1±5.3 vs. 16.8±7.8)(P=0.001 vs. P=0.001), the GCS score was significantly higher after treatment(13.9±4.4 vs. 12.8±4.2)than be-fore treatment(9.5 ± 3.8 vs. 9.6 ± 3.7)(P=0.001 vs. P=0.021). The symptomatic intracranial hemorrhage rate was signifi-cant higher in anterior circulation group (5/24, 20.8%) than in posterior circulation group (0,0%) (P<0.05). Compared with the anterior circulation group, the recanalization rate trended to increase in posterior circulation group (P=0.830).The symptomatic intracranial hemorrhage rate in anterior circulation group (5/24,20.8%) was significant higher than in pos-terior circulation group (0,0%) (P<0.05), the mortality was similar between these two groups. Conclusions Intra-arterial thrombolysis combined with mechanical adjuvant therapy can improve neurological deficit in acute large artery occlusive infarction and increase the recanalization rate,which is more suitable for the treatment of posterior circulation infarction.

Objective To investigate the morbidity rate of hyperuricemia and its association with hypertriglyceridemia and hypertension in elderly people in Fujian Quanzhou.Methods A crosssectional population survey for hyperuricemia was performed in Fujian Quanzhou.Questionnaire and physical examination were conducted in 1358 subjects.The levels of serum UA and lipid-profile as well as blood pressure were measured.Results In the same aged group,the level of blood uric acid was significantly higher in male (371.7±83.6) μmol/L than in female (294.8±66.5) μmol/L (t=15.8,P＜0.01).Blood uric acid concentration was gradually increased with aging in the group at age 40-70 years,but was gradually decreased with aging in female group aged over 70 years (F=12.1,P＜0.01).The total morbidity rate of hyperuricemia was 18.6％.The incidence of hypertriglyceridemia or hypertension in female with hyperuricemia was higher than those with normouricemia (22.1％ vs.10.5％,59.1％ vs.26.2％,x2=16.3,65.0,all P＜0.01).The incidence of hypertension in male with hyperuricemia was higher than those with normouricemia (54.6％ vs.42.8％,x2=4.0,P＜0.05).Conclusions The morbidity rate of hyperuricemia in elderly people is higher in south China coast than other region of China.The incidence of hypertriglyceridemia and hypertension in people with hyperuricemia is significantly higher than those with normouricemia.

ObjectiveTo elucidate the clinical application value of intravascular ultrasound(IVUS) in assessing intermediate lesions and the relationship between serum levels of fractalkine,CD36 and LP-PLA2and severity of coronary artery atherosclerosis.MethodsOne hundred and twenty patients with unstable angina underwent coronary angiography.Then the patients were divided into two groups:severe lesion group (lumen diameter stenosis greater than 70％,including 40 patients),and the intermediate lesion group lumen diameter stenosis(50％～70％,80 patients).IVUS were performed in all patients to analysis the coronary lesions.Concentrations of CD36,fractalkine and LP-PLA2 were measured by means of ELISA.Results IVUS found that 74％ intermediate lesion patients had soft lipid plaques,while 48％ severe lesion patients had lipid plaques ( P＜0.01 ).Patients with severe lesion had larger plaque burden and vascular remodeling index than those of patients with intermediate lesions(P＜0.05～0.01).There were 59 patients(74％) in the intermediate lesion group underwent PCI.IVUS also found the PCI patients had larger plaque burden and vascular remodeling index.Concentration of CD36 was significantly higher in severe lesion group than that of intermediate lesion group (P＜0.05).ConclusionsIVUS could further clarify the characteristics of the intermediate and severe lesions,and provide guidance for PCI treatment.Plasma CD36 level could be used to predict pathological severity of coronary atherosclerosis.

OBJECTIVETo study the effect of Ginkgo biloba extract on rats during ischemia/reperfusion and its influence on intracellular calcium in hippocampal neurons.

METHODSModel of intraluminal occlusion of the middle cerebral artery (MCAO) was used to prepare the ischemia/reperfusion cortex tissue. Concentration of MDA was determined by measuring thiobarbituric acid-reactive substance. GSH-PX was quantified using the thiobarbituric acid (TBA) technique. SOD was assayed througha xanthine method. Endogenous amino acids were quantified by high performance liquid chromatographic (HPLC) analysis. Primary culturs of hippocampal neurons were prepared for a free intracellular calcium ([Ca(2+)]I) assay by Fura-2 based single cell microfluoremetric technique.

RESULTSComparing control and treatment groups, the concentration of SOD and GSH-PX were higher, whereas that of MDA was much lower; the concentration of glutamate and aspartate decreased and that of GABA increased markedly at all time point (P < 0.01), Gly also decreased at some time points (P < 0.05). The differences were significant between the groups of 10 mg/kg, 15 mg/kg and the groups of 5 mg/kg. When 1 x 10(-5) mol/L glutamate was applied with 25 micro g/ml ginkgo biloba extract to cultured neurons, the increase in [Ca(2+)]I was lower than that caused by applying glutamate alone. Its peak value was much lower and increased phase was longer, its declining phase was shorter. After returning to baseline, the application of 1 x 10(-5) mol/L glutamate could induce the reaction to recover.

CONCLUSIONSGinkgo biloba extract could protect damaged neurons by keeping the balance of inhibitory/excitatory aminoacids, enhancing the free radical scavengers system, and inhibiting the effect of glutamate on [Ca(2+)]I.

Amino Acids ; analysis ; Animals ; Brain Ischemia ; metabolism ; Calcium ; metabolism ; Cells, Cultured ; Free Radicals ; Ginkgo biloba ; Hippocampus ; metabolism ; Lipid Peroxidation ; drug effects ; Male ; Neuroprotective Agents ; pharmacology ; Phytotherapy ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Reperfusion Injury ; prevention & control

Although bulk endocytosis has been found in a number of neuronal and endocrine cells, the molecular mechanism and physiological function of bulk endocytosis remain elusive. In pancreatic beta cells, we have observed bulk-like endocytosis evoked both by flash photolysis and trains of depolarization. Bulk-like endocytosis is a clathrin-independent process that is facilitated by enhanced extracellular Ca(2+) entry and suppressed by the inhibition of dynamin function. Moreover, defects in bulk-like endocytosis are accompanied by hyperinsulinemia in primary beta cells dissociated from diabetic KKAy mice, which suggests that bulk-like endocytosis plays an important role in maintaining the exo-endocytosis balance and beta cell secretory capability.
Animals ; Calcium ; metabolism ; Cytoplasmic Granules ; metabolism ; Diabetes Mellitus ; metabolism ; pathology ; Disease Models, Animal ; Dynamins ; metabolism ; Electric Capacitance ; Endocytosis ; physiology ; Insulin ; metabolism ; Insulin-Secreting Cells ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Patch-Clamp Techniques ; Photolysis ; Primary Cell Culture

Upon glucose elevation, pancreatic beta-cells secrete insulin in a Ca(2+)-dependent manner. In diabetic animal models, different aspects of the calcium signaling pathway in beta-cells are altered, but there is no consensus regarding their relative contributions to the development of beta-cell dysfunction. In this study, we compared the increase in cytosolic Ca(2+) ([Ca(2+)]i) via Ca(2+) influx, Ca(2+) mobilization from endoplasmic reticulum (ER) calcium stores, and the removal of Ca(2+) via multiple mechanisms in beta-cells from both diabetic db/db mice and non-diabetic C57BL/6J mice. We refined our previous quantitative model to describe the slow [Ca(2+)]i recovery after depolarization in beta-cells from db/db mice. According to the model, the activity levels of the two subtypes of the sarco-endoplasmic reticulum Ca(2+)-ATPase (SERCA) pump, SERCA2 and SERCA3, were severely down-regulated in diabetic cells to 65% and 0% of the levels in normal cells. This down-regulation may lead to a reduction in the Ca(2+) concentration in the ER, a compensatory up-regulation of the plasma membrane Na(+)/Ca(2+) exchanger (NCX) and a reduction in depolarization-evoked Ca(2+) influx. As a result, the patterns of glucose-stimulated calcium oscillations were significantly different in db/db diabetic beta-cells compared with normal cells. Overall, quantifying the changes in the calcium signaling pathway in db/db diabetic beta-cells will aid in the development of a disease model that could provide insight into the adaptive transformations of beta-cell function during diabetes development.
Animals ; Calcium ; metabolism ; Calcium Signaling ; drug effects ; Cell Membrane Permeability ; drug effects ; Cells, Cultured ; Down-Regulation ; drug effects ; Endoplasmic Reticulum ; metabolism ; Glucose ; pharmacology ; Insulin-Secreting Cells ; cytology ; drug effects ; metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Obese ; Potassium Chloride ; pharmacology ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; metabolism ; Sodium-Calcium Exchanger ; metabolism ; Thapsigargin ; pharmacology ; Up-Regulation ; drug effects