Objective To investigate the predictive value of serum microRNA-506 (miR-506) and microRNA-934 (miR-934) expression levels in postoperative survival outcomes of patients with pancreatic cancer. Methods A total of 115 patients with pancreatic cancer who received surgical treatment were selected as the study group, and were divided into survival group (n=18) and death group (n=97) according to the follow-up results. Fifty patients with benign pancreatic lesions in the same period were selected as control group; fifty healthy volunteers were selected as the healthy group. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the relative expression levels of miR-506 and miR-934 in serum of each group. Multivariate Logistic regression analysis was used to analyze the influencing factors of postoperative survival outcomes in patients with pancreatic cancer. The predictive value of serum miR-506 and miR-934 in postoperative survival outcome of pancreatic cancer patients was analyzed by receiver operating characteristic (ROC) curve. Results The expression level of serum miR-506 in the study group was significantly lower than that in the control group (P < 0.05). The expression level of serum miR-934 in the study group was significantly higher than that in the control group and healthy group (P < 0.05). The expression level of serum miR-506 in the survival group was significantly higher than that in the death group, and the expression level of serum miR-934 was significantly lower than that in the death group (P < 0.05). The miR-506, miR-934, TNM stage and lymph node metastasis were independent factors influencing postoperative survival of pancreatic cancer patients (P < 0.05). Conclusion The expression of miR-506 is low in the serum of patients who died after pancreatic cancer surgery, and the expression of miR-934 is high in the serum of patients who died after pancreatic cancer surgery. The value of miR-506 and miR-934 in predicting postoperative survival outcome of pancreatic cancer patients is high.

Objective:To investigate the effect of autophagy on liver injury with obstructive jaundice in Sprague-Dawley (SD) rats and its underlying mechanism.Methods:Thirty-five healthy male SD rats, SPF grade, aged 6-8 weeks, weighting 200-300 g, were divided into 5 groups with 7 rats in each group, including sham group (simple free common bile duct, without ligation, intraperitoneal injection of normal saline), obstructive jaundice (OJ) group (established by common bile duct ligation, intraperitoneal injection of normal saline), OJ group with 3-MA, OJ group with Rapamycin, and OJ group with 3-MA and VX-765. Morphological changes in liver tissues were analyzed with HE staining. Expression of autophagy-related protein Atg5 was detected by immunohistochemistry staining. Liver function was analyzed by automatic biochemical instrument and the level of serum interleukin (IL)-18 was detected using ELISA assay. Protein levels of autophagy related-proteins and endoplasmic reticulum stressed (ERs)-related apoptosis proteins were detected by Western Blot.Results:The relative expression of autophagy related protein Atg5 in OJ group was significantly higher than that in sham group [(5.0±1.0) vs. (2.8±1.3), t=-3.00, P<0.05]. Compared with sham group, the activity of autophagy was enhanced and the protein levels of Caspase-1/p-65 and IL-18 were significantly increased in OJ group. At the same time, apoptosis was induced by activating ERs. In OJ group, the autophagy inducer 3-MA improved the expression levels of Caspase-1/p-65 and IL-18, and aggravate liver injury. While after applying the autophagy agonist Rapamycin in OJ rat models, the expression of Caspase-1/p-65 and IL-18 was repressed and liver damage was also reduced. In addition, in rat OJ groups with 3-MA, inhibition of Caspase-1 by VX-765 could down regulate the expression of Caspase-1/p-65 and IL-18, and protect against liver injury. Conclusions:Both ERs related apoptosis and autophagy were activated after ligation of common bile duct. Besides, activation of autophagy could reduce OJ-induced liver injury in SD rats by inhibiting the Caspase-1/p-65 inflammatory pathway.

Objective:To investigate the effect of cell migration-inducing hyaluronidase 1 (CEMIP) on biological function of gallbladder cancer GBC-SD cells and its possible mechanism.Methods:The expression of CEMIP in biliary epithelial cell line HIBEC and gallbladder cancer cell line NOZ and GBC-SD was detected by Western blot. GBC-SD cells in logarithmic growth phase were divided into blank control group, negative control group (transfection with nonsense siRNA), siCEMIP-1 group (transfection with siCEMIP-1) and siCEMIP-2 group (transfection with siCEMIP-2). The expression of CEMIP and binding immunoglobulin protein (Bip) and calreticulin (CRT) in GBC-SD cells was detected by Western blot after culturing for 48h in blank control group, negative control group, siCEMIP-1 and siCEMIP-2 group. The relative survival rate was determined by CCK-8 assay. The wound healing rate and apoptotic rate was detected by wound healing assay and flow cytometry. The migration and invasion abilities were evaluated by Transwell chamber assay. Twelve 5-week-old BALB/c-nude mice were selected and divided into control group and experimental group (6 mice in each group). GBC-SD cells and GBC-SD cells with silenced CEMIP were subcutaneously injected into the right armpit (forelimb) of the two groups of mice, respectively. The volume and weight of transplanted tumor were compared 33 days later.Results:Compared with HIBEC cells, the relative protein level of CEMIP in gallbladder cancer GBC-SD cells [(0.750±0.034) vs. (0.120±0.002)] and NOZ cells [(0.690±0.013) vs. (0.120±0.002)] was significantly higher ( P<0.05). Compared with blank control group and negative control group, the relative protein level of CEMIP, Bip and CRT in siCEMIP-1 group and siCEMIP-2 group was significantly lower ( P<0.05). Compared with blank control group and negative control group, the relative survival rate and wound healing rate and number of migration cells and invading cells in siCEMIP-1 group and siCEMIP-2 group were also significantly lower ( P<0.05). While the apoptotic rate in siCEMIP-1 group and siCEMIP-2 group were higher than that in blank control group and negative control group ( P<0.05). Compared with control group, the average tumor volume [(543.6±114.7) vs. (801.5±256.3) mm 3] and tumor weight [(0.453±0.093) vs. (0.728±0.278) g ] of the experimental group was significantly decreased ( P<0.05). Conclusions:CEMIP was up-regulated in gallbladder cancer cell line GBC-SD and NOZ. Silencing CEMIP inhibited cell proliferation, wound healing rate, migration and invasion, and promoted apoptosis in gallbladder cancer GBC-SD cells, which may be related to the inhibition of endoplasmic reticulum chaperone Bip and CRT expression.

Hepatocellular carcinoma (HCC) with bile duct tumor thrombus (BDTT) is rare and enhanced CT or MRI can be used for its diagnosis. Surgical procedure is the main treatment for HCC with BDTT. The authors introduce the experiences of recurrent patient with HCC and BDTT who was treated with targeted therapy plus immunotherapy, in order to provide reference for its clinical diagnosis and treatment.

Objective:To determine the clinical efficacy of selective decongestive devascularization of gastrosplenic (SDD-GSR) and splenectomy combined with pericardial vascularization in the treatment of portal hypertension in cirrhosis.Methods:A total of 134 patients with cirrhosis portal hypertension admitted to the First Affiliated Hospital of Wenzhou Medical University were enrolled in the study, including 102 males and 32 females, with an average age of 51 years. Of 61 cases of SDD-GSR were included in the SDD-GSR group, and 73 cases of splenectomy combined with pericardial vascularization were included in the control group. Preoperative and postoperative white blood cell count, platelet count, Child-Pugh grade of liver function, free portal pressure (FPP) and postoperation tomplication were analyzed in the two groups. Operation time, intraoperative blood loss, free portal pressure (FPP), Child-Pugh grade of liver function, preoperative and postoperative white blood cell count, platelet count, and postoperative complications were analyzedin the two groups.Results:The operation time and intraoperative blood loss of SDD-GSR group were 165 (110, 198) min and 280 (100, 650) ml, which were lower than those of control group [190 (135, 605) min and 895 (300, 3 500) ml], the differences were statistically significant ( P<0.05). Postoperative FPP of SDD-GSR group and control group was 39 (35, 44) cmH 2O (1 cmH 2O=0.098 kPa) and 38 (34, 44) cmH 2O, respectively, which were lower than those before operation, with statistical significance (both P<0.05). Postoperative platelet count and white blood cell count in SDD-GSR group were lower than those in control group, and the differences were statistically significant (all P<0.05). The Child-Pugh grading of recent postoperative liver function in SDD-GSR group was better than that in control group, with statistical significance ( P<0.05). The complication rate (abdominal infection and portal vein thrombosis) of control group was higher than SDD-GSR group. Conclusion:SDD-GSR is better than splenectomy combined with pericardial vascularization since it has less intraoperative bleeding, obvious improvement of liver function and fewer complications, and it may be an effective surgical option for the treatment of portal hypertension of cirrhosis.

【Objective】 To investigate the expression of pyruvate kinase M2 (PKM2) in intrahepatic cholangiocarcinoma (ICC) and the effect of PKM2 on the proliferation and epithelial-to-mesenchymal transition (EMT) of ICC cells. 【Methods】 PKM2 expression was evaluated in ICC tissues and cell lines by Real-time quantitative reverse-transcription polymerase chain reaction (RT-PCR), and immunohistochemistry assays. In vitro, we knocked down PKM2 expression in ICC cell lines and investigated the biological function and the underlying mechanism of PKM2 in ICC. 【Results】 RT-PCR results showed that PKM2 was highly expressed in ICC (P<0.05). Moreover, PKM2 knockdown inhibited cell proliferation and the invasive capacities of ICC cells, and inhibited Wnt/ β-Catenin signaling, which subsequently regulated EMT signaling pathway in vitro. 【Conclusion】 PKM2 expression in cholangiocarcinoma is significantly higher than that in adjacent tissues. PKM2 can regulate EMT and invasion and metastasis of ICC via the Wnt/β-Catenin signal.

Objective:To report a rare case of paroxysmal nocturnal hemoglobinuria (PNH) complicated with chronic tubulointerstitial nephropathy, combined with literature review, and discuss the clinical, imaging and pathological characteristics of the disease and the diagnosis and treatment ideas.Methods:The patient's clinical data, magnetic resonance imaging (MRI) and kidney pathological examination results, treatment measures and effects were collected and reported. Through systematic review of relevant literature, the clinical manifestations and pathogenesis of chronic tubular interstitial nephropathy complicated by PNH were summarized and discussed.Results:In this case, PNH was diagnosed for more than 30 years, the peripheral blood PNH clone was positive, urine specific gravity was 1.012, urine pH 6.0-7.0, urine protein (+), urine sugar (3+), serum creatinine 259 μmol/L, serum lactic acid dehydrogenase 800 U/L. MRI showed bilateral renal cortical signal was low intensity on both T1- and T2- weighted images. Kidney biopsy revealed remarkable chronic tubulointerstitial nephropathy with massive hemosiderin deposition in proximal tubular cells demonstrated by Prussian blue staining and electron microscopy. By using low-dose prednisone to control hemolytic attack and other supportive treatments, the patient's renal function has been stabilized for a long time.Conclusions:PNH complicated with chronic tubulointerstitial nephritis is easy to be misdiagnosed due to insidious onset. MRI and kidney histopathological examination are helpful to clarify the diagnosis. Early diagnosis and treatment are helpful to improve the prognosis of such patients.

Objective: To explore the clinical efficacy of S-1 single agent adjuvant chemotherapy for the patients undergoing radical resection of extrahepatic biliary carcinoma. Methods: The clinical data of 108 patients with extrahepatic biliary carcinoma receiving radical resection who were admitted from January 2014 to June 2017 were retrospectively analyzed. There were 62 males(57.4%)and 46 females(42.6%),with a median age of 59 years (range:26 to 79 years),10 cases(9.3%) in stage Ⅱ,85 cases(78.7%) in stage Ⅲ, and 13 cases (12.0%) in stage Ⅳ, 40 cases(37.0%) of hilar cholangiocarcinoma, 8 cases(7.4%) of middle cholangiocarcinoma, 25 cases (23.2%) of distal cholangiocarcinoma, 35 cases(32.4%) of gallbladder carcinoma.After radical resection of extrahepatic biliary carcinoma, 49 patients receiving S-1 single agent chemotherapy and 59 patients receiving non-special treatment were divided into the chemotherapy group and the operation group,respectively. All the dates of the patients were followed up and collected with the overall survival time,tumor-free survival time,1,2 and 3-year survival rate after operation,and the rate of major toxic reaction during chemotherapy of the chemotherapy group. Survival curve was drawn by the Kaplan-Meier method, and survival analysis was done using the Log-rank test. Results: There were no significant differences in the general date of two groups(sex, age, tumor size, tumor site, TNM stages, degree of differentiation). The median overall survival time and the median tumor-free survival time in the chemotherapy group were 27 months and 21 months,respectively,and in the operation group were 21 months and 17 months,respectively. There were differences between the two groups in the overall survival rates(χ²=3.967,P<0.05) and the 2 and 3-year survival rate(63.3%,36.6%;41.6%,20.4%;χ²=4.510,P<0.05;χ²=6.143,P<0.05),but the 1-year overall survival rate (83.4%,79.7%)was not statistically significant(χ²=0.286,P>0.05). There were no significant differences in the tumor-free survival time,1,2 and 3-year tumor-free survival rate(77.6%,41.4%,33.1%;62.7%,30.9%,21.2%)between the two groups(χ²=0.876,P>0.05;χ²=0.252,P>0.05;χ²=1.571,P>0.05;χ²=3.323,P>0.05,respectively). The main toxic reaction during chemotherapy were dyspepsia(28.6%, 14/49), anemia(26.5%, 13/49), and leukopenia(22.5%, 11/49), all of which were mild. Conclusion S-1 single agent chemotherapy after radical reseetion of extrahepatic biliary carcinoma could effectly improve the survival of patients and all of the main toxic reaction during chemotherapy were mild.

Objective: To explore the effect of aspirin combined with metformin on the apoptosis of thyroid cancer TPC-1 cells and its mechanism. Methods: The proliferation and apoptosis of TPC-1 cells treated with different concentrations of aspirin and metformin were detected using cell count kit-8 (CCK-8) assay and flow cytometry, respectively. Western blot was used to detect the expressions of microtubule-associated protein light chain 3 (LC3), p62 and cysteinyl aspartate specific proteinase 3 (caspase-3) after treatment with aspirin, metformin and 3-Methyladenine (3-MA). Results: The relative cell viability of TPC-1 cells treated with 0.5, 1.0, 2.0, 4.0 mmol/L aspirin for 24 and 48 hours were (85.6±9.1)%, (79.9±8.6)%, (57.0±5.3)%, (55.7±5.4)%; (76.7±2.8)%, (75.4±6.1)%, (46.1±4.1)%, (36.3±3.2)%, respectively. The value of half maximal inhibitory concentration (IC₅₀) for 24 and 48 hours were 4.297 mmol/L, 2.133 mmol/L, respectively. The apoptotic rate in the 1 mmol/L aspirin treatment group and negative control group were (29.2±8.5)%, (4.2±2.9)%, respectively (P<0.05). Moreover, treatment with metformin increased the protein expression of LC3Ⅱ/Ⅰ ratio, and decreased the expression of p62, while treatment with aspirin decreased the expression of LC3Ⅱ/Ⅰ ratio and increased the expression of p62. The relative cell viability of TPC-1 cells treated with metformin, 3-MA, an autophagy inhibitor, and 3-MA combined with metformin were (73.2±9.2)%, (95.8±3.3)%, (59.9±9.2)%, respectively. The apoptotic rates in these groups were (35.5±1.5)%, (12.3±1.4)%, (49.9±5.4)%, respectively. Compared with the metformin group, the relative cell viability in metformin combined with 3-MA group was significantly lower while the apoptotic rate was higher (P<0.05), which indicated that treatment with 3-MA enhanced the metformin-induced apoptosis of TPC-1 cells. The relative cell viability of TPC-1 cells in metformin group, aspirin group, metformin combined with aspirin group were (87.3±11.8)%, (85.7±9.6)%, (72.4±8.8)%, respectively. The apoptotic rates in these groups were (29.7±4.0)%, (30.5±6.5)%, (52.5±4.6)%, respectively. Compared with the metformin or aspirin group, the relative cell viability in metformin combined with aspirin group was significantly lower, while the apoptotic rate was higher (P<0.05), which indicated that aspirin enhanced the metformin-induced apoptosis of TPC-1 cells. Conclusions Our findings indicate that metformin-mediated autophagy plays a protective role in metformin-induced apoptosis and proliferation inhibition. Aspirin enhances the metformin-induced apoptosis of thyroid cancer TPC-1 cells through inhibition of autophagy.

Objective To explore the effect of aspirin combined with metformin on the apoptosis of thyroid cancer TPC?1 cells and its mechanism. Methods The proliferation and apoptosis of TPC?1 cells treated with different concentrations of aspirin and metformin were detected using cell count kit?8 ( CCK?8) assay and flow cytometry, respectively. Western blot was used to detect the expressions of microtubule?associated protein light chain 3 (LC3), p62 and cysteinyl aspartate specific proteinase 3 ( caspase?3) after treatment with aspirin, metformin and 3?Methyladenine ( 3?MA). Results The relative cell viability of TPC?1 cells treated with 0.5, 1.0, 2.0, 4.0 mmol／L aspirin for 24 and 48 hours were ( 85.6 ± 9.1)%, (79.9±8.6)%, (57.0±5.3)%, (55.7±5.4)%; (76.7±2.8)%, (75.4±6.1)%, (46.1±4.1)%, (36.3±3.2)%, respectively. The value of half maximal inhibitory concentration ( IC50 ) for 24 and 48 hours were 4.297 mmol／L, 2.133 mmol／L, respectively. The apoptotic rate in the 1 mmol／L aspirin treatment group and negative control group were (29.2±8.5)%,(4.2±2.9)%, respectively (P＜0.05). Moreover, treatment with metformin increased the protein expression of LC3Ⅱ／Ⅰ ratio, and decreased the expression of p62, while treatment with aspirin decreased the expression of LC3Ⅱ／Ⅰ ratio and increased the expression of p62. The relative cell viability of TPC?1 cells treated with metformin, 3?MA, an autophagy inhibitor, and 3?MA combined with metformin were (73.2±9.2)%, (95.8±3.3)%, (59.9±9.2)%, respectively. The apoptotic rates in these groups were (35.5±1.5)%, (12.3±1.4)%, (49.9±5.4)%, respectively. Compared with the metformin group, the relative cell viability in metformin combined with 3?MA group was significantly lower while the apoptotic rate was higher ( P＜0.05), which indicated that treatment with 3?MA enhanced the metformin?induced apoptosis of TPC?1 cells. The relative cell viability of TPC?1 cells in metformin group, aspirin group, metformin combined with aspirin group were ( 87.3 ± 11.8)%, ( 85.7 ± 9.6)%, ( 72.4 ± 8.8)%, respectively. The apoptotic rates in these groups were ( 29.7± 4.0)%, (30.5 ± 6.5)%, ( 52.5 ± 4.6)%, respectively. Compared with the metformin or aspirin group, the relative cell viability in metformin combined with aspirin group was significantly lower, while the apoptotic rate was higher ( P＜0.05), which indicated that aspirin enhanced the metformin?induced apoptosis of TPC?1 cells. Conclusions Our findings indicate that metformin?mediated autophagy plays a protective role in metformin?induced apoptosis and proliferation inhibition. Aspirin enhances the metformin?induced apoptosis of thyroid cancer TPC?1 cells through inhibition of autophagy.