AIM To investigate the roles of Cl- channels in Ca2+ influx induced by activaion of al- adrenoceptor subtypes in transfected-CHO cells. METHODS The effects of drugs on ?1A、?1B and ?1D- AR-induced Ca2+ influx were investigated with Fura2 fluorescence technique. RESULTS The ?1A-AR- induced Ca2+ influx was inhibited by furosemide(2 .5 ~ 10 M?mol?L- 1 )and SK&F96365(5- 15 ?mol?L- 1 ) in a concentration- dependent manner respectively; The ?1B-AR-induced Ca2+ influx could also be inhibit inhibited by NFA(2. 5 ~ 10 ?mol? L-1 ), whereas the alD AR-induced Ca2+ influx was only suppressed by NFA. In ?1B-CHO cells, Adr-triggered Ca2+ influx could be further inhibited by NFA or furosemide after the maximal inhibition by SK&F96365;SK&F96365 could further inhibit Ca2+ influx which had been inhibited by NFA or furosemide. In ?1A-CHO cells, Adr-triggered Ca2+ influx could be further inhibited by SK&F96365 after had been inhibited by furosemide; furosemide could not further inhibite Ca2+ influx which had been inhibited by S&F96365. CONCLUSION There are different characteristics of CI- channels related to ?1A、 ?1B and ?1D-AR-induced Ca2+ influx.

Objective To investigate the incidence trends,clinical features and prognosis of ischemic colitis (IC) in China,and improve the level of diagnosis and treatment of IC.Methods China National Knowledge Infrastructure and VIP Chinese Science and Technology Periodical Database were searched.The ending date of search was May 15,2014.Results A total of 324 references were found,which were published from 1982 to 2013.There were 9202 cases reported,3973 cases of males and 5229 cases of females,with the male/female ratio of 1 ∶ 1.32 and mean age of (63.6±7.8)years.The amount of references and cases reported began to rise after 2002.Estimated ratio of cumulative incidence was higher in the north than in other areas of China (x2 =1097.95,P=0.000).The most common IC accompanying diseases were hypertension,heart diseases,diabetes,hyperlipidemia and constipation.There were statistically significant differences in the accompanying diseases between different regions and different times.Drugs,enteroscopy,surgery and low blood volume might be the precipitating factors.Patients commonly complained of abdominal pain,diarrhea/ desiring to defecate and hematochezia.Computed tomography was feasible in detecting lesions.Colonoscopy was the main method for diagnosis.The lesions were most common located in the left half colon including sigmoid colon,descending colon and splenic flexure,with typically in a segmental manner.Pan-colon involvement or rectum involvement rarely occurred.Type of transient lesion was the predominant subtype,which was generally managed non-operatively with good prognosis.Different from the type of transient lesion,pathological changes in gangrenous type were located in the right half colon including hepatic flexure of transverse colon,ascending colon and ileocecal junction.Gangrenous type required prompt surgical intervention but the mortality was much higher.Conclusions Incidence of CI has been increasing year by year.Patients who complain of abdominal pain,diarrhea/desiring to defecate and hematochezia should be considered as IC,particularly in the elderly women and patients with cardiovascular disease.Diseased regions are mainly located in sigmoid colon,descending colon and splenic flexure.Transient colitis is the predominant subtype,which have good prognosis.

AIM: To investigate the relationship between chloride channels and the Ca 2+influx induced by adrendine(Adr). METHODS: The effects of drugs on Adr-induced Ca 2+influx were investigated with Fura-2 fluorescence technique. RESULTS: Adr-induced Ca 2+influx was inhibited by nifedipine,SK&F96365,NFA and furosemide in a concentration manner respectively; Ca 2+influx could be further inhibited by NFA or furosemide after the maximal inhibition by SK&F96365;SK&F96365 also could further inhibit the Ca 2+influx which had been inhibited by NFA or furosemide. Genistein and vanadate could reduce or increase the Ca 2+influx respectively. CONCLUSION: Ca 2+influx induced by Adr is related to VDC and ROC, and chloride channels involves in the processes.The levels of tyrosine phosphoralation affect the Ca 2+influx.

To comparatively investigate ultrastructural characteristics and expressions of AFP (alpha-fetoprotein) and Tn (Thomsen-Friedenreich-related antigen) protein in AFP negative (AFP-) and AFP positive (AFP+) primary hepatocellular carcinoma. Fourty-three cases of AFP- and AFP+ hepatocellular carcinoma (HCC) tissues and five cases of normal liver tissues were divided into three groups: control group (normal liver tissue, n=5); AFP+ HCC group (the serum AFP level was higher than 10 ng/ml, n = 22); AFP- HCC group (the serum AFP level was lower than 10 ng/ml, n=21). The ultrastructural morphology was studied by transmission electron microscopy, the expressions of AFP and Tn protein were detected by immunohistochemistry and cell image analysis. 1. The immunohistochemical study showed that (1) the expression intensity and positive rate of Tn protein in AFP- HCC group were markedly higher than that in AFP+ HCC group (P<0.01); (2) The expression intensity of AFP in AFP- HCC group was lower than that in AFP+ HCC group (P<0.01). 2. The transmission electron microscopy demonstrated that some AFP- HCC cells linked closely with each other, others dispersed loosely just as cultured cells, the remarkable morphologic features in AFP- HCC cells were simple organelles, but they were abundant in the free polyribosomes. In AFP+ HCC group, all the HCC cells linked closely together and were rich organelles in their cytoplasm, especially the rough endoplasmic reticula. In addition, mitochondria and Golgi complex were obviously observed. (1) The AFP and Tn protein had discrepancy distribution in AFP- and AFP+ HCC tissues, Tn protein may be one of the early diagnostic indicators in AFP- HCC; (2) The synthetic locations of the AFP and Tn protein were different in hepatocarcinoma cells by ultrastructural observation.

The effects of drugs on intracellular calcium concentration(［Ca2+］i) were investigated with fura-2 fluorescence technique to investigate ATP and thrombin-induced Ca2+ entry in bovine aortic endothelial cells(BAEC). It was found that application of ATP and thrombin gave rise to biphasic ［Ca2+］i elevation. ATP or thrombin only triggered a fraction of cyclopiazonic acid(CPA)-sensitive Ca2+ store, which was enough to activate Ca2+ entry. The Ca2+ release induced by thrombin resulted from the activation of phospholipase C(PLC), whereas the PLC-independent mechanism was involved in ATP-induced Ca2+ release. Nifedipine had no effect on ATP and thrombin- induced Ca2+ entry. SK&F 96365 and ginsenoside-2A inhibited both ATP and CPA-induced Ca2+ entry, however no effect of them on thrombin-induced Ca2+ entry was found. The inhibitory effects of SK&F 96365 and ginsenoside-2A on CPA-induced Ca2+ entry were less than that on ATP-induced Ca2+ entry. The Ca2+ influx sensitive to SK&F 96365 was not the same as that to ginsenoside-2A. These observations suggest that both ATP and thrombin evoke Ca2+ release and Ca2+ influx by activation of different receptor. However their mechanisms appear different.

Objective To evaluate the value of indirect hemagglutination test(IHA)in schistosomiasis diagnosis. Meth?ods The literature concerned schistosomiasis diagnosis with IHA in the databases of Medline,CNKI,VIP and Wanfang Data from 1982 to 2014 was collected and evaluated. Results Totally 21 articles which were satisfied with the research criteria were analyzed with the Meta?analysis method. The IHA method had high value in schistosomiasis diagnosis,the AUCSROC of IHA in laboratory evaluation was 0.990 6,while in filed evaluation was 0.832 9,and the difference between them was significant(Z=4.50,P<0.05). Conclusion The diagnosis value of IHA in field evaluation is less than that in laboratory. In the process of the elimination of schistosomiasis,developing a new and higher sensitive reagent in schistosomiasis diagnosis is needed.

Objective:This study aimed to investigate the effect of human lymphatic endothelial cells (HLECs) on proteins secreted by epithelial ovarian cancer (EOC) cells SKOV3-pm4 with highly directional lymphatic metastasis. Methods:The supernatants of the four groups of cultured cells (A, SKOV3;B, SKOV3+HLEC;C, SKOV3-PM4;and D, SKOV3-PM4+HLEC) were collected. The proteins of these cells were detected by antibody arrays and iTRAQ-2D-LC-MALDI-TOF/TOF/MS. The screened significantly differential proteins were further analyzed by bioinformatics and validated in the human serum and cell culture medium by ELISA. Results:Progranulin (GRN) and vascular endothelial growth factor A (VEGF-A) were upregulated between groups C and A. In addition, insulin-like growth factor binding protein-7 (IGFBP-7) and secreted protein acid rich in cysteine (SPARC) were downregulated between groups D and C. Comprehensive bioinformatics analysis revealed that IGFBP7 interacted with VEGFA. VEGF exhibited the highest expression in ovarian cancer and IGFBP7 exhibited the lowest expression compared with the serum of the normal control group. Statistically significant differences were observed between the two substances. Conclusion:The HLEC microenvironment is closely associated with directional metastasis in lymph nodes with differential proteins, including matricellular proteins and adhesion factors. In particular, the upregulation of VEGFA and GRN and the downregulation of SPARC and IGFBP7 were closely associated with the directional metastasis of EOC cells in lymph nodes.

Objective To determine the effect of signal transducers and activators of transcription 3 (STAT3) gene silencing by shRNA mediated by lentiviral vector for the treatment of colorectal cancer. Methods The recombinant lentiviral vector pRNAT-shSTAT3, empty lentiviral vector pRNAT-GFP, and lentiviral packaging plasmids in supernatant were collected to transfect HT-29 cells for harvesting the HT-29-shSTAT3 cells and HT29-GFP cells. Fifteen male rats were divided into three groups (n = 5 ), and then they were inoculated with HT-29cells, HT-29-GFP cells and HT-29-shSTAT3 cells, respectively. Cell growth was assessed by MTT assay and the changes in cell cycle were detected by flow cytometry. The changes in microvessel density (MVD) of tumors were detected by immunohistochemistry. All data were analysed by one-way analysis of variance. Results The growth of HT-29-shSTAT3 cells was significantly suppressed compared with HT-29 and HT-29-GFP cells (F = 632.50,P ＜ 0. 05 ). The proportions of cells at the G0/G1 phase were 68.7% ± 2.9% in HT-29-shSTAT3 cells, 38.5% ±1.6% in HT-29-GFP cells and 38.7% ± 2.3% in HT-29 cells, with a significant difference among the three groups (F = 166.53, P ＜ 0.05 ). The MVDs of HT-29 cells, HT-29-GFP cells and HT-29-shSTAT3 cells were 29 ±5, 28 ±4 and 10 ±3, respectively, with a significant difference among the three groups (F=31.60, P ＜0.05). Conclusion STAT3 gene silencing by shRNA mediated by lentiviral vector can significantly inhibit the growth of colorectal cancer cells.

Objective To establish a human ovarian carcinoma cell line with directional highly lymphatic metastasis and to study their biological characteristics.Methods The clone cells of ovarian carcinoma,SKOV3,were inoculated into the hind foot pad of nude mice.The cancer cells of lymph node metastatic foci were transplanted into nude mice again when the metastatic nude of mice were observed.After repetition of this procedure for 3 cycles,the metastatic rate and the metastatic paths were observed in nude mice of every passage.We used limited dilution method to separate and select colonial cells with directional highly lymphatic metastatic potentials from the lymphatic metastasis of human ovarian carcinoma cell line SKOV3.The ceils with biological characteristics were assayed by growth curve,HE staining,karyotype analysis,nude mice transplantation and immunohistochemistry,respectively.Results We established a series of cell lines from lymph node metastasis and designated them as SKOV3-PM1,SKOV3-PM2 and SKOV3-PM3 cell strain.When the cells of SKOV3-PM3 were injected into the hind foot pad of nude mice, they produced 100%(10/10)spontaneous lymphatic metastasis.The lymphatic metastatic rates(26/10) were stable and higher than the mother cell line(1/10,P

Objective To compare anatomically the potential incidences of injury to the nerve roots of ventral and dorsal rami caused by the transarticular screws and Magerl lateral mass screws in the fixation of subaxial cervical spine.Methods Fixations with the transarticular screws and Magerl lateral mass screws were performed in this study.For each kind of fixation,20 screws were inserted from C3 to C7 in two specimens.A 20-ram long screw was used to over-penetrate the ventral cortex.The anterolateral aspect of the cervical spine was carefully dissected to allow ob- servation of the screw-ramus relationship.Results The overall percentage of nerve violation was significantly lower in the transarticular screw fixation(45%)than in the Magerl lateral mass screw fixation(90%).The difference was sta- tistically significant between the two fixation methods(P＜0.05).The greatest percentages of nerve violation caused by the transarticular screws and lateral mass screws were found at the dorsal ramus,25% and 45%,respectively. Conclusion The potential risk of nerve root violation caused by the transarticular screw technique is lower than that by the Magerl lateral mass screw technique in the lower cervical spine.