Objective To investigate the value of digital image analysis (DIA) of endoscopic ultrasonography (EUS) in the differential diagnosis of benign and malignant pancreatic neuroendocrine neoplasm (PNEN).Methods The relation between various parameters of EUS-imaging and benign and malignant lesions in 47 patients clinically diagnosed PNEN were retrospectively analyzed.Photoshop CS5 software was performed for digital image processing,and lesions related parameters were collected,including area,perimeter,length,circularity,gray,gray ratio,and gray standard deviation.The statistical method of t test was performed for comparison between two groups.Receiver operating characteristic (ROC) curve was analyzed in length,circularity,average gray scale ratio and gray standard deviation.Results Among the 47 patients,35 cases and 12 cases were in benign group and malignant group,respectively.The mean gray scale ratio and the circularity of benign group were significantly higher than those of malignant group (0.80±0,05 vs 0.74±0.07,0.63±0.17 vs 0.40±0.09),and the differences were statistically significant (t=2.659 and 5.787,both P＜0.05).The gray standard deviation of benign group were lower than that of malignant group (9.90 ± 1.24 vs 12.55± 3.27),and the difference was statistically significant (t=-2.733,P=0.018).The area under the curve (AUC) of circularity was 0.724 (95％ confidence interval(CI):0.546 to 0.901),the cut-off value was 0.767,the sensitivity and specificity were 71.43 ％ and 66.67％,respectively.The AUC of average gray ratio was 0.888 (95％CI:0.785 to 0.991),the cut-off value was 0.412,the sensitivity and specificity were 94.29％ and 75.00％,respectively.The AUC of gray standard deviation was 0.811 (95％CI:0.647 to 0.974),the cut-off value was 11.02,the sensitivity and specificity were 66.67％ and 85.71％,respectively.When combined with the three parameters of circularity,average gray scale ratio and gray standard deviation,the sensitivity and specificity were 97.14％ and 83.33％,and the accuracy was 93.61％.Conclusions EUS with DIA technology can improve the detection of EUS images to PNEN,and which may be complementary to EUS guided fine needle aspiration.It also privided a noninvasive,objective,convenient,and effective option for the differential diagnosis of benign and malignant PNEN.

Objective To assess the clinical value of low-frequency mini - probe sonography ( LFMPS) in preoperative localization of pancreatic endocrine tumors comparing with other imaging methods. Methods Twenty one cases with suspected pancreatic endocrine tumors were enrolled from June 2000 to June 2002, we compared the diagnostic results of LFMPS, transcutaneous ultrasonography ( US) , helico-computed tomography ( HCT) and magnetic resonance imaging (MRI) with surgical localization and histopathological results by using Fujinon 7. 5 MHz miniature probe and SP-701 ultrasonic system. Results Sixteen pancreatic insulinomas and 1 extra pancreatic VIPoma (vesoactive intestinal polypeptide tumor) were confirmed by surgery and histopathological examination in 17 of the 21 patients, and the rest 4 patients didn't receive surgical procedure because of the negative results in all imaging studies. Among pancreatic lesions, they located on head, body and tail in 9, 3 and 4 cases respectively; the average diameter of all 17 lesions was 2. 02cm. LFMPS correctly localized the tumor in 14 of 17 patients (82. 4% ) while CT in 15 of 17 patients (88. 2% ) , MRI in 12 of 17 patients (70. 6% ) and US in 9 of 17 patients (52. 9% ). Besides, the diagnostic accuracy of LFMPS in detection of small size (

Objective To explore the relationship between microRNA (miRNA)-200c expression,epithelial-mesenchymal transition (EMT) and the sensitivity to gefitinib in colon cancer cells.Methods The inhibitory effects of gefitinib on four types of colon cancer cell lines (HT29,SW620,HCT1116,SW480) were examined by cell counting kit-8 (CCK-8) assay.The expressions of miRNA-200c,epithelial marker (E-cadherin) and mesenchymal markers (vimentin and zinc finger Ebox binding homeobox 1 ZEB 1) at mRNA level in four types of colon cancer cell lines were detected by fluorescence quantitative polymerase chain reaction.The expressions of E-cadherin,vimentin and ZEB 1 at protein level were determined by Western blot.After up-or down-regulated the expression of miRNA-200c,the changes of the expression of EMT related genes and the sensitivity to gefitinib were observed.Results HT29 cells were most sensitive to gefitinib (IC50 =(7.70 ± 0.31) μmol/L),in which the expressions of both miRNA-200c and E-cadherin were the highest,and the expressions of vimentin and ZEB1 were extremely low.HCT116 and SW480 were moderately sensitive to gefitinib (IC50=(11.88±0.97) and (16.63±0.45) μmol/L),and the expressions of miRNA-200c and Ecadherin were moderate.SW620 cell line was most insensitive to gefitinib (IC50 =(26.43 ± 3.68)μmol/L),the expressions of miRNA-200c and E-cadherin were the lowest.The expressions of vimentin and ZEB 1 in SW620 were higher than that of the other three types of cell lines.After upregulated the expression of miRNA 200c,the expression of E-cadherin in SW620 cells increased,the expression of ZEB 1 and vimentin decreased,and the sensitivity to gefitinib increased.After downregulated the expression of miRNA-200c,the expression of E-cadherin in HT29 cells decreased,the expression of ZEB 1 and vimentin increased,and the sensitivity to gefitinib decreased.Conclusion miRNA-200c may up-regulate the expression of E-cadherin through EMT regulation,and then influence the sensitivity to gefitinib in colon cancer cells.

Objective To investigate the effects of interferon-α (IFN-α) and gefitinib on the proliferation and apoptosis of human colon cancer cell line HCT116. Methods Colon cancer cell line HCT116 was selected as research objective. The biological effects of IFN-α and gefitinib alone or combined on the cells were observed at different time point (after worked for 24, 48 and 72 hours). The proliferation inhibition of the medicine on the HCT116 cells was measured by methyl thiazolyl tetrazolium (MTT) assay. Morphologic changes were observed under optical microscope. Apoptosis was measured by flow cytometry (FCM). The results were analyzed with SPSS 13.0 software, two groups compare was tested by t test, and single factor variance analysis was for multiple group data compare. Results IFN-α and gefitinib alone or combined could significantly inhibit the proliferation of HCT116 cells (P<0.05), and there was a time and dose-dependent manner between the degree of inhibition and the working time and concentration of the medicine. With the work of the medicine, apoptosis morphologic changes were observed in the cells. And FCM result indicated that the apoptosis rate significantly increased. After treated with IFN-α and gefitinib alone or combined for 72h, the cell apoptosis rate were 15.6%±0.6%, 13.6%±0.4% and 31.2%±0.3% respectively, which was obviously higher than control group (6.8%±0.3%, P<0.05). Conclusion Both IFN-α and gefitinib were able to inhibit the proliferation and induce apoptosis of HCT116 cells moreover, and a synergistic effect was observed while combine used there two medicines.

Objective To investigate the effect of antioxidants including PDTC on pancreatic fibrosis of rats with chronic pancreatitis.Methods The rats were randomly divided into 5 groups including control group, CP group, PDTC treatment group, vitamin E treatment group and vitamin C treatment group.The CP model was in ducad by using intraperitoneal injection of DETC (750 mg/kg), twice a week.The control group received no treatment.After DETC injection, the treatment groups received an intraperitoneal injection of PDTC (100 mg/kg), vitamin E (15 mg/kg), vitamin C (15 mg/kg), respectively.Rats were sacrificed at 90 min, 24 h, 48 h, 72 h, 2 w, 3 w, 4 w, 6 w after first injection of DETC.Pancreatic tissue was taken for routine pathological examination.The activity of SOD, GSH-PX and MDA content were detected by spectrophotometric ratio method.α-SMA, desmin collagen Ⅰ, Ⅲ, TGF-β1, FN were detected by immunohistochemical assay.The expression of TGF-β1, FN mRNA was measured by RT-PCR.Results At 6w, the fibrosis and the parameters for damage of the pancreas in the three treatment groups were significantly better than that in CP group (P ＜0.01), the vacuolar degeneration index in vitamin E group and vitamin C group was also better than that in CP group (P ＜0.01).From the 2nd week, the activity of SOD, GSH PX in PDTC group, Vit C group and Vit E group was higher than that in CP group, while the MDA activity was lower than that in CP group, and the difference was statistically significant (P ＜ 0.01 or P ＜ 0.05).No significant difference was found among the three treatment groups.The mRNA levels of TGF-β1 and FN of the treatment groups were lower than those of CP group (P ＜0.05 or P ＜0.01), but higher than those of the control group (P ＜ 0.05).There was no significant difference among the three treatment groups (P ＞ 0.05).Conclusions PDTC and the other antioxidants can reduce oxygen free radicals by increasing the activity of SOD,suppressing the activation of PSCs, reducing the secretion of TGF-β1, Collagen Ⅰ , Ⅲ, FN and eventually inhibit the progress of pancreatic fibrosis.

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.

The paper analyzed legally the following predicaments of legitimization of rural doctors in China:no legitimacy protection for their practice,vague criteria for practicing medicine,conflicts between legal regulations for medical practice and rural realities,and lack of a liability insurance system.The authors,based on an exploration of the legislative framework for rural doctors,made four corresponding legislative proposals as follows:to clearly define the legal scope and identity of rural doctors,to establish a qualification and employment system for rural doctors,to develop practicing norms applicable for rural doctors,and to set standards for their income.

Objective To investigate the effect of protein inhibitor of activated signal transducer and activator of transcription 1 ( PIAS1 ) gene silencing on the inflammatory response of rat pancreatic acinar cell lines AR42J with cerulein stimulation, to study its role in the pathogenesis of acute pancreatitis.Methods The siRNA targeting PIASI was designed, synthesized, transfected into AR42J cells by lipofectmine 2000.24 h later, cerulean was added and cultured for another 24 h.Subsequent AR42J cells with cerulein stimulation were divided into 4 groups: cerulein, liposome, negative-siRNA and PIAS1-siRNA groups.In addition, a group with PBS was as control group.The activity of p38 mitogen- activated protein kinase (p38MAPK) was detected by western blotting.TNF-α, IL-1β, IL-6, matrix metalloproteinase (MMP) 9 expression were analyzed by RT-PCR and western blotting, respectively.Results The expression of p38MAPK in PIAS1-siRNA, negative-siRNA, liposome, cerulein,and control group was 1.93 ±0.11, 1.22 ±0.10, 1.30 ±0.17,1.32 ± 0.21, 0.12 ± 0.02;while the expression of phosphorylated p38MAPK was 2.10 ± 0.25, 1.36 ± 0.20,1.26 ±0.15, 1.23 ±0.25, 0.58 ±0.48, the expression in PIAS1-siRNA group was significantly increased when compared with other groups (P＜0.05).The levels of TNF-α, IL-1β, IL-6, MMP-9 mRNA were 1.66 ±0.15,1.66 ± 0.15,1.90 ±0.01, 1.56 ±0.20 in PIAS1-siRNA group, while the expression of protein was 2.06 ±0.37,2.20 ±0.34, 1.80 ±0.10, 1.17 ±0.05, which was markedly higher than those in other group (P ＜0.05).Conclusions PIAS1 gene silencing could enhance p38MAPK activity, and improve inflammatory mediator expression in pancreatic acinar cells with cerulein stimulation.

Objective To study the value of EUS-FNA cytology and fluid carcinoembryonic antigen (CEA) for differential diagnosis of malignant and benign pancreatic cystic lesions.Methods Data of 27 patients who underwent EUS-FNA were reviewed.According to Youden exponent,the optimal cut-off points for cyst fluid CEA were determined by receiver operating characteristic (ROC) curve.Compared with surgical pathology,the accuracy,sensitivity,specificity,positive predictive value (PPV) and negative predictive value (NPV) of the EUS imaging,cytology as well as cyst fluid CEA were determined.Results Of the 27 cases,14 were diagnosed as benign lesions,13 were diagnosed as malignant or premalignant lesions.The accuracy,sensitivity,specificity,PPV and NPV of EUS imaging were 77.8％ (21/27),69.2％(9/13),85.7％ (12/14),81.8％ (9/11) and 75.0％ (12/16).The accuracy,sensitivity,specificity,PPV and NPV of EUS-FNA cytology were 85.2％ (23/27),76.9％ (10/13),92.9％ (13/14),90.9％ (10/11),and 81.3％ (13/16).The corresponding values of fluid carcinoembryonic antigen under the ROCderived ideal cut-off were 74.1％ (20/27),84.6％ (11/13),64.3％ (9/14),68.8％ (11/16) and 81.8％ (9/11) (CEA ＞ 22.24 ng/ml).Conclusion EUS-FNA cytology is highly accurate and specific for differential diagnosis of malignant and benign pancreatic cystic lesions.Cyst fluid CEA shows better sensitivity.EUS-FNA cytology and cyst fluid CEA analysis can basically meet the requirement of differentiating the benign and (pre)malignant pancreatic cystic lesions.

Objective To evaluate the diagnostic value of endoscopic ultrasound guided fine needle aspiration (EUS-FNA) in combination with flow cytometry (FCM) in lymphoma.Methods From January 2011 to December 2011,the cases of suspicious lymphoma with EUS-FNA examination at Shanghai Ruijin Hospital were retrospectively analyzed.The final diagnosis was according to pathological diagnosis of specimen from the surgery and follow up results.The sensitivity,specificity and accuracy of EUS-FNA combined with FCM in lymphoma diagnosis were initially analyzed.Results A total of 14 suspicious lymphoma patients were collected,eight cases were diagnosed as lymphoma by pathological examination of specimen from the surgery or.tissue from aspiration,four cases were non-lymphoma lesions and two cases still had no final diagnosis.The sensitivity and specificity of FCM alone in lymphoma diagnosis were 4/8 and 4/4 respectively.Six cases of lymphoma were detected by EUS-FNA with FCM.The sensitivity,specificity and accuracy of EUS FNA combined with FCM were 6/8,6/6 and 10/12 respectively.Conclusion EUS-FNA combined with FCM has better diagnostic value in lymphoma,especially for gastrointestinal lymphoma and those surrounding deep lesions.