Objective To explore the diagnostic values of bile cytology obtained in endoscopic retrograde cholangiopancreatography (ERCP) and blood tumor markers for malignant bile duct obstruction.Methods From August 2005 to April 2009, bile cytology and blood tumor markers measurement were performed in 47 cases with bile duct obstruction, in which 39 cases were confirmed to be malignant and 8 were benign. Results Malignant cells were found in bile from 26 of 39 malignant obstruction (66. 7% ), and serum CA19-9 was positive in 28 (71.8%). No malignant cells were detected in bile cytology from 8 cases with benign obstruction (with a specificity of 100% ) and serum CA19-9 was negative in 7 (87.5%). In 26 patients with cholangiocarcinoma, serum CA19-9 was positive in 18 (69. 2% ). For bile cytology and blood tumor markers test, sensitivity of parallel combination was 89. 7%, and the specificity of serial combination was 100. 0%. Conclusion Bile cytology testing during ERCP can provide the pathological evidence for malignant bile duct obstruction. The sensitivity is improved by combination tests of bile cytology and blood tumor markers.

Objective To study the effects of Irinotecan (CPT-11) on human colorectal cancer HCT-116 and HT-29 cells and investigate the potential mechanisms.Methods The effect of Irinotecan on the proliferation of HCT-116 and HT-29 cells was determined by MTT assays.The invasive capacity was measured by transwell assays,and the apoptosis of the tumor cells was detected by flow cytometry after stained with Annexin-V and PI.The difference between the current of ATP-sensitive potassium ion of HCT-116 and HT-29 was examined by patch clamp.Results It was found that 1.0-64.0 μg/ml CPT-11 could inhibit the proliferation and the invasive capacity of HCT-116 and HT-29 cells at both dose-and time-dependent manner.The IC_(50) of HCT-116 and HT-29 were 39.3 and 19.5 μg/ml respectively.Cytometry showed that the apoptotic rates were increased from 14.8% and 9.3% to 36.9% and 27.9% after the treatment of 32.0 μg/ml and 16.0 μg/ ml CPT-11,which were close to their IC_(50).The proportion of G_0/G_1 and S of HCT-116 and HT-29 was enhanced from 27.4% and 17.4% to 95.9% and 98.2%.Transwell assay indicated that the invasiveness of HCT-116 and HT-29 was reduced by 40.8% and 47.5%.The patch clamp showed that CPT-11 reduced the I_(KATP) of cell membrane at a negative dose-dependent manner.Conclusion CPT-11 could have a significant impact on the proliferation,invasiveness,cell cycle,and the apoptosis of human colorectal cancer cell HCT-116 and HT-29.HT-29 was more sensitive to CPT-11 than HCT-116.The inhibitory effect of CPT-11 on cell proliferation might be linked to its inhibition of ATPsensitive potassium channel.

Objective To investigate the effects and potential mechanism of irinotecan (CPT-11), an antitumor drug, on human colorectal cancer cell line HT-29 and its impact on 4-amion pyridine (4-AP), a kalium ion channel blocker. Methods The effects of CPT-11, 4-AP and combination of two drugs on proliferation and invasion of HT-29 cells were measured by MTT and Transwell assay respectively. The impact of CPT-11 or 4-AP on cell apoptosis was determined by flow cytometry with Annexin-V and PI staining. The current of ATP sensitive potassium ion (IKATP) was measured by patch clamp. Results The CPT-11 could inhibit proliferation of HT-29 cells at dose from 1.0 to 64.0 μg/ml in dose-and time-dependent manners. Whereas the above effect was enhanced when CPT-11 combined with 4-AP (1.0 mmol/L). The administration of CPT-11 (16.0 μg/ml) or 4-AP (1.0 mmol/L) significantly induced the cell apoptosis and inhibited the invasion of HT-29 cells, furthermore, these effects could be enhanced by combination of two drugs. And the different concentrations of CPT-11 reduced the IKATP of cell membrane in negative dose-dependent manner. Conclusions The effects of CPT-11 on HT-29 cells, such as reducing proliferation and invasion as well as inducing the apoptosis, can be enhanced by 4-AP, which may be related to inhibition of ATP-sensitive potassium channels.

Objective To assess the related risk factors of endoscopic retrograde cholangiopancreatography(ERCP)on postoperative pancreatitis.To improve the level of diagnostic and therapeutic ERCP,to reduce the incidence of postoperative pancreatitis.Methods A total of 346 patients in our hospital referred to diagnostic and therapeutic ERCP(399 frequency)were divided into 8 groups;the differences of postoperative serum amylase in 24 hour as well as clinical symptoms were compared among different groups.Results The incidence of postoperative hyperamylasaemia was 12.5 %.The incidence of postoperative acute pancreatitis was 1.5 %.ERCP+STENT therapeutic group(38.9%)had the highest incidence of postoperative hyperamylasaemia and postoperative acute pancreatitis among the groups.(respectively 38.9%,11.1%).There was the different incidence of postoperative actcte pancreatitis between the period from June 2003 to June 2005 and from July 2005 to July 2007,respectively 3.3%,0.4%.Conclusion Pancreatic duct contrast filling and deficient experience of doctor during ERCP manipulation are the main risk factors for postoperative pancreatitis.

Objective To investigate the pathologic changes in the pancreas of rats after intraperitoneal injection of DETC,a kind of superoxide dismutase (SOD) inhibitor,and to compared that with another model of chronic pancreatitis by pancreatic duct injection of TNBS.Methods The rats were randomly divided into DETC group,DETC control group,TNBS group,TNBS control group,normal control group.Rats in DETC group received an intra-peritoneal injection of DETC twice a week,and rats in DETC control group received an intra-peritoneal injection of same amount of normal saline.Rats in TNBS group was injected with 2％ TNBS ethanol phosphate buffer into the pancreatic duct,while rats in TNBS control group was treated with injection of same amount of ethanol phosphate buffer,and rats in normal control group received no treatment.The rats were sacrificed after 2 w,4 w,6 w and 8 w.The serum levels of amylase were determined,and pathological and ultrastructure changes of the pancreas were measured.The levels of SOD,GSH-PX activity and MDA content were detected.The expressions of α-SMA,Desmin,Collagen Ⅰ,Collagen Ⅲ,TGF-β1,FN in tissue were detected by immunohistochemical assay.The TGF-β1 mRNA expression was detected by RTPCR.Results No rat died in DETC group.The mortality rate of TNBS group was 15％.The serum levels of amylase were not statistically different between the 2 groups.The fibrosis scores of rat in DETC group at 4 w was 3.4 ± 1.l,which was significantly higher than that in TNBS group (3.0 ± 1.3,t =3.462,P ＜ 0.05).At 6 w,the damage scores of rat in DETC group was 9.1 ± 1.8,which was significantly higher than that in TNBS group (8.4 ± 1.8,t =2.943,P ＜ 0.05).Scores of vacuolar degeneration and fatty infiltration of rat in DETC group were higher than those in TNBS group,but the difference between the two groups was not statistically significant.Two weeks later,ultrastructure changes of pancreas could be observed,and large amounts of regenerative or mature collagen could be seen at 4 w.The SOD activity of DETC group was significantly decreased when compared with those in TNBS group (t =5.468,P ＜ 0.01).The GSH-PX activity of DETC group at 2 w,6w was significantly decreased when compared with those in TNBS group (t =6.497,10.125,P＜0.01).While the activity of MDA at 6 w,8 w was significantly increased when compared with those in TNBS group (t =3.350,5.407,P ＜0.05).The differences at other time points were not statistically significant.The expressions of (a)-SMA,Desmin,Collagen Ⅰ,Collagen Ⅲ,TGF-β1,FN,and TGF-β1 mRNA were not statistically significant between the 2 groups.Conclusions Sustained suppression of SOD activity can successfully induce chronic pancreatitis.Fatty infiltration and fibrosis in pancreas in DETC group occurs earlier with more severe presentation than that in TNBS group.Intraperitoneal injection of DETC is easy with low mortality rate,which is an ideal method for chronic pancreatitis model induction.

Objective:Our purpose was to investigate the growth regulation and postreceptor signal transduction of somatostatin (SS) on human colon cancer cell line. Methods:Low differentiated clone A cells of human carcinoma were treated with different concentrations of SS and the growth status was observed. Intracellular 1,4,5-trisphosphate (IP3) and cyclic adenosine monophosphate (cAMP) were extracted, then the concentrations of them were measured by liquid scintillation technique and gamma scintillation counter.Intracellular free calcium concentration was detected by loading Fura-2 and fluorescental technique. Protein kinase C (PKC) was extracted from cytosol and membrane, then the activity of both parts was determined with TaKai method. Results:The clone A cell growth was inhibited greatly by different concentrations of SS (10-10 to 10-5　mol/L) and it is related to the doses of SS.Somatortatin could largely inhibit the production of intracellular IP3, ［Ca2+］i, and cAMP, and decrease the activity of PKC. Conclusion:The growth of Clone A cell can be inhibited by SS. The inhibition may be mediated by phosphate inositol pathway, so intracellular IP3, ［Ca2+］i decreased, or inhibited the cell growth by inhibiting the activity of PKC.On the other hand, the cell proliferation may be inhibited by adenosine cyclase pathway, that is decreasing intracellular cAMP ,inhibiting cAMP-depending protein kinase.

Objective To evaluate pit pattern analysis for detection of early colorectal carcinoma and precancerous lesions. Methods A total of 162 lesions in 144 patients were examined with magnifying colonoscopy after staining, and their pit patter was analyzed with morphology and pathologic diagnosis. Results With confirmation of pathology, there were 34 non-neoplastic lesions and 128 neoplastic ones, in which 12 were carcinomas. The pit patterns in most non-neoplastic lesions (76. 5%, 26/34) were type Ⅰ or Ⅱ , and those in most neoplastic lesions (96. 1% , 123/128) was type Ⅲ, Ⅳ or Ⅴ. Pit patterns of cancerous lesions were mainly type Ⅴ (75.0%, 9/12), and those of 3 cases of advanced cancers were all type Ⅴ N. Conclusion Pit pattern classification is a very important tool to differentiate between neoplastic, nonneoplastic lesions and early cancer, which helps to decide later therapeutic intervention.

Objective To identify the association between human glucocorticoid receptor gene G166T polymorphism and essential hypertension(EH).Methods DNA samples from 71 normotensive cases and 48 EH cases were analyzed by a polymerase chain reaction(PCR) for restriction fragment length polymorphism(RFLP) to determine the intron 4 variant of human glucocorticoid receptor gene.Results No association was observed between EH group and control group.However,in females,a little association was observed between the EH group and control group.Frequencies of allele G were 0.64 in EH group and 0.46 in control group.Conclusion The G allele may be a predisposing gene marker,HGR gene intron 4 polymorphism contributes to the development of EH in females.

Objective To evaluate the clinical application value of anti-β2 glycoprotein I ( anti-β2 GPI) and anti-cardiolipin antibodies ( ACA ) in the patients with antiphospholipid syndrome ( APS).Methods Serum levels of anti-β2GPI(IgG) and ACA(IgA, IgG, IgM) were determined by enzyme linked immunosorbent assay ( ELISA ) in 53 patients with APS , 27 patients with SLE accompanied by APS , 55 patients with simple SLE , 46 patients with other autoimmune diseases and 40 healthy controls.The sensitivity and specificity of anti-β2 GPI and ACA for the diagnosis of APS and the correlation of serum anti-β2 GPI and ACA-IgG levels were analyzed.The cases were identified in the Affiliated Hospital of Qingdao University during 2012.1 to 2014.2 and the data were analyzed with χ2 test, Mann-Whitney U test and Spearman correlation.Results The positive rates and serum levels of anti-β2 GPI, ACA-IgG/M, ACA-IgG, ACA-IgM were significantly higher in the APS group [77.4%(41/53), 81.1%(43/53), 56.6%(30/53), 52.8%(28/53);14.1 AU/ml, 19.6 U/ml, 17.9 U/ml] than those in the simple SLE group [16.4%(9/55), 32.7%(18/55), 20.0%(11/55), 18.2%(10/55); 4.9 AU/ml, 9.4 U/ml, 8.7 U/ml, χ2 =40.4, 25.7, 15.4, 14.2;U=255.0, 632.5, 476.5, P<0.01], other autoimmune diseases group[0%(0/46), 4.3%(2/46), 2.2%(1/46), 2.2%(1/46); 3.2 AU/ml, 2.6 U/ml, 3.4 U/ml, χ2 =60.7, 58.6, 33.9, 30.5;U=53.5, 87.0, 66.0, P<0.01] and healthy controls [0%(0/40), 2.5%(1/40), 0%(0/40), 2.5%(1/40);3.0 AU/ml, 3.5 U/ml, 2.9 U/ml, χ2 =55.3, 56.5, 33.4, 26.9;U=61.0, 124.0, 152.0, P <0.01 ] separately.The positive rate and serum level of ACA-IgA in the APS group [18.9%(10/53), 11.7 U/ml] were significantly higher than that in the other autoimmune disease group [2.2%(1/46), 2.9 U/ml,χ2 =6.9, U=581.0, P<0.01] and healthy controls[2.5%(1/40),2.1 U/ml,χ2 =4.4,U=764.0,P<0.05] separately.The specificity of anti-β2 GPI (83.6%) for APS diagnosis was significantly higher than that of ACA (67.3%, χ2 =4.0,P<0.05).Conclusions anti-β2 GPI, ACA-IgG and ACA-IgM, have higher clinical application value in the APS diagnosis and identification of SLE with or without APS than ACA-IgA.The specificity of anti-β2 GPI for APS diagnosis is higher than that of ACA.

Objective To investigate the protective effect of RLN revealed for RLN in high -risk thyroid surgery,and the clinical value of detecting nerve,thus to provide a reference for clinical treatment.Methods 132 patients of thyroid high -risk surgery were selected,according to whether to explode recurrent laryngeal nerve,they were divided into exposed group (80 cases)and non -exploded group(52 cases).The recurrent laryngeal nerve damage rate,clinical indicators of operation time were compared between the two groups.In the explode group,80 patients according to whether implementation of the nerve monitoring,were divided into the monitored group (36 cases)and non -monitored group(44 cases).The hours to find the nerve,recurrent laryngeal nerve injury rates and other indicators were compared.Results The operative time of the exploded group was (133.66 ±21.48)min,which was significantly longer than the non -exploded group,difference was statistically significant (t =3.994,P =0.025 ).The exposed group had similar incidence of hypocalcemia compared with the other groups,there was no statistically significant difference (P >0.05).The revealing recurrent laryngeal nerve injury rate of the exposed group was 2.50%,which was significantly lower than the non -exploded group,difference was statistically significant (χ2 =5.794,P =0.008).The recurrent laryngeal nerve positioning time of the monitoring group was (4.78 ±0.94)min,which was significantly shorter than the unmonitored group,difference was statistically significant (t =7.168,P =0.001).The recurrent laryngeal nerve injury rate of the monitoring group was 2.78%,showed no significant difference (χ2 =0.911,P =0.338).Conclusion In a high risk thyroid surgery,explode recurrent laryngeal nerve can reduce the rate of recurrent laryngeal nerve damage,nerve monitoring can effectively speed up the nerve localization time,but has no significant effect on reduce the rate of recurrent laryngeal nerve injury.