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The expression of miR-101 in pancreatic cancer and its effect on proliferation of pancreatic cancer cell line ASPC1

Xianpeng LI ; Shiwei GUO ; Zhendong JIN ; Le QU

Chinese Journal of Pancreatology.2010;10(4):276-278. doi:10.3760/cma.j.issn.1674-1935.2010.04.016

Objective To investigate the expression of miR-101 in pancreatic cancer and the effect of down-regulation miR-101 on proliferation of pancreatic cancer cell line ASPC1. Methods Real-time PCR was used to determine the expression of miR-101 in pancreatic cancer, adjacent tissues and pancreatic cancer cell line ASPC-1. The miR-101 over-expression vector (peGFPc1-miR-101) was constructed and was transfected into ASPC-1 cell. Transfection efficiency was measured by fluorescence microscope. The expression of miR101in the transfected cells was detected by real-time PCR. Cell viability analysis was performed by MTT. The targeted genes of miR-101 in pancreatic cancer were scanned by the online targeted gene prediction software (target Scan). Results The expression of miR-101 was in pancreatic cancer tissues, adjacent tissues and ASPC-1 cell line, respectively. The expressions in pancreatic cancer tissues and ASPC-1 cells were significantly lower than that in adjacent tissues ( P ＜ 0.01 ). The expression of miR 101 in transfected cells increased to 19.8 folds as much as that in the control group (P ＜0.01 ). Proliferation rate of transfected cells was significantly decreased, which was only 26% of primary cells ( P ＜ 0.01 ). EZH2 was the potential targeted gene of miR-101 in pancreatic cancer. Conclusions miR-101 was weakly expressed and it may affect the proliferation of pancreatic cancer cell by inhibiting the EZH2 expression.

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In vitro anti-tumor effect of autologous mixed lymphocyte primed by BCG activated dendritic cells based PANC1 lysate

Dehong YANG ; Wenjia LIU ; Min CHEN ; Yumei WU ; Xiaoping ZOU

Chinese Journal of Pancreatology.2010;10(4):267-271. doi:10.3760/cma.j.issn.1674-1935.2010.04.014

Objective To evaluate in vitro anti tumor effect of host lymphocyte primed by CalmetteGuerin bacillus (CGB) activated dendritic cells (DC) based PANC1 lysate. Methods DCs were obtained from peripheral blood mononuclear cells of healthy volunteer and cuitured by rhGM CSF and rhIL 4. DC vaccines for pancreatic cancer were loaded with PANC1 tumor lysate (TL) and were further maturated by CGB.CD1a, CD83, CD86 and HLA-DR phenotype was characterized by flow cytometer, and IL-12p70 and TNF-α concentration in DC culture supernatant were measured by ELISA. Autologous mixed lymphocyte proliferation and the cytotoxicity of cytotoxic T lymphocytes primed by activated DCs to PANC1, PaTu8988 and SCG7901 tumor cells was measured by CCK 8 test. Results When DCs based PANC1 lysate were activated by CGB,the expression rates of CD83 and CD86 were increased from (3.7±0.3)% and (38.6±5.0)% to (16.5±0.6)% and (76.6±2.5)% (P ＜0.05 ). The concentrations of cytokines ILpl2p70 and TNF-α were increased from (20.18±2.06 ) pg/mland (61.38±1.19) pg/mlto (62.48±3.80) pg/mland (592.53±17.96)pg/ml (P＜0. 01 ). When co-cultured with CGB activated DCs based PANC1 lysate in proportion of 0.40)% , (3.39±1.05)% , (2.82±0.39)% significantly increased to (55.38±3.58)% , (75.0±2.54) % , (77.07±3.4)% , (99.07±2.4)% (P＜0.01) , respectively. The killing effects of lymphocytes 2.77)%, (19.03±3.04) %; but the killing effects on PaTu8988 and SCG7901 were significantly decreased.Conclusions DC vaccines for pancreatic cancer could be more maturated when activated by CGB, and could show a high capability of anti-tumor in vitro.

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Separate purification and directed differentiation potency of pancreatic stem cells of adult rats in vitro

Guangjun SHI ; Lei ZHANG ; Ping XU ; Jiang YU

Chinese Journal of Pancreatology.2010;10(4):260-262. doi:10.3760/cma.j.issn.1674-1935.2010.04.012

Objective To investigate the experimental condition of separate purification and directed differentiation potency of pancreatic stem cells of adult rats in vitro. Methods The pancreas of adult rats were digested by collagenase V in situ perfusion. Filtration was performed by 100 mesh filter. Ficoll 400 density gradient centrifugation was used to separate pancreatic stem cells and stem cells were for cultivation. Epidermal growth factor (EGF), fibroblast growth factor (bFGF) was added to medium for in vitro cultivation.Immunofluorescence staining method was used to detect the expression of CK19, Pdx-1, nestin, insulin and glucagon, and the positive cell rate was calculated. The differentiated cell was evaluated by dithizone (DTZ)staining; insulin excretion function was determined by optical enzyme labeled ELISA staining. Results The pancreatic stem cell obtained from the study could be cultivated in vitro for 8 generations. The expression of CK19, Pdx-1 and nestin of the tem cells were all positive, and the rate of positive cells were (88.6 ± 6.2)% ,(84.6 ±8.6)% and (81.3 ±7.5)%. The differentiated cell was in brownish red color by DTZ dye. After high concentration sugar stimulation, the expression of insulin secretion was increased in the supernatant.Conclusions This method can harvest highly purified, and large amount of pancreatic duct stem cell.Artificial induction may result directed differentiate for islet-like clusters and have insulin secrete function.

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Chemotherapy of pancreatic cancers by drug sensitivity test in vitro

Qinghua LIU ; Li ZHANG ; Minghua ZHU

Chinese Journal of Pancreatology.2010;10(4):257-259. doi:10.3760/cma.j.issn.1674-1935.2010.04.011

Objective To investigate the effect of cancer drug sensitivity on the selection of chemotherapy for pancreatic cancer. Methods The cells from the cancer tissues of 156 pancreatic cancer patients were cultured with 6 kinds of chemotherapy drugs in vitro including gemcitabine (GEM), 5 fluorouracil (5-FU), Mitomycin C (MMC), Oxaliplatin (L-OHP) and irrinotecan (CPT-11), according to the in vitro standard of solitary tumor. More than 70% of inhibitory rate was highly sensitive, 50% ～ 70% was moderately sensitive, ＜ 50% was insensitive. The inhibitory rate of chemotherapy drugs were determined by MTT colorimetric assay. Results The pathological findings of the 156 cases of pancreatic cancer were pancreatic duct cancer in 135 cases, adenosquamous carcinoma in 13 cases, mucinous carcinoma in 8 cases.Pancreatic duct cancer was sensitive to all the 6 drugs; adenosquamous carcinoma and mucinous carcinoma was sensitive to all the drugs except for L-OHP and CPT-11, respectively. The inhibitory rate of GEM was higher than that of MMC, L-OHP and CPT-11 (P ＜ 0.05 ) , but there was no difference with 5-FU and DDP ( P ＞0.05 ). There was no difference among the other 5 drugs (P ＞ 0.05 ). However, the cells from different types of pancreatic cancers and the cells from different patients of the same type of pancreatic cancer have different sequence of sensitivity to the 6 kinds of chemotherapy drugs. For pancreatic duct cancer, the sequence of sensitivity was GEM ＞ DDP ＞ 5-FU ＞ CPT-11 ＞ MMC ＞ L-OHP; adenosquamous carcinoma was GEM, CPT-11 ＞ DDP, 5-FU, MMC ＞ L-OHP; mucinous carcinoma was L-OHP ＞ GEM ＞5-FU, MMC ＞ DDP ＞ CPT-11.Conclusions Cancer drug sensitivity test may help to select the fight chemotherapy and be of clinical value.

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Clinical value of serum CA19-9 levels in preoperative evaluation of resectability of pancreatic carcinoma

Guang LI ; Shun ZHANG

Chinese Journal of Pancreatology.2010;10(4):251-253. doi:10.3760/cma.j.issn.1674-1935.2010.04.009

Objective To evaluate the value of serum CA 19-9 levels in predicting resectability of pancreatic cancer. Methods Serum CA 19-9 levels of 52 patients with pancreatic cancer who were possible to be curatively resected according to the imaging were determined; the gold standard was respectability. An receiver operating characteristic (ROC) curve was constructed for the CA 19-9 levels; the point that lies closest to the upper left-hand comer of the graph is chosen as the cut-off point, the sensitivity, specificity,positive and negative predictive values were calculated. Results There were 29 (55.8%) resectable and 23 (44.2%) inoperable cases of pancreatic cancers. The CA19-9 level in resectable patients was (159.6±170.9)U/ml, and it was (944.4 ±773.4) U/ml in inoperable cases. The area under the ROC curve was 0. 918 ( ＞ 0.9, P ＜ 0.01 ), 95% confidence interval were 0.843 ～ 0.992. The CA19-9 level was (353.2) U/ml according to the cut-off point, the sensitivity and specificity was 93.1% and 78.3%, respectively; positive and negative predictive values was 84. 4% and 90. 0% , respectively. Conclusions Preoperative serum CA19-9 level may be a useful marker for further evaluating the respectability of pancreatic cancer who are possible to be resected according to the imaging studies.

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Diagnosis and treatment of the periampullary carcinoma in the pre-jaundice stage

Shaowu HE ; Gang JIN ; Xiangui HU ; Junsheng KANG

Chinese Journal of Pancreatology.2010;10(6):398-400. doi:10.3760/cma.j.issn.1674-1935.2010.06.006

Objective To explore the clinical characteristics of periampullary carcinoma in the prejaundice stage and improve early diagnosis rate and operative effects.Methods Clinical data of 27 cases with periampullary carcinoma in the prejaundice stage in our institute during the period of Jan 1998 to Dec 2005were analyzed retrospectively.Results The clinical symptom was mostly nonspecific, mainly included abdominal discomfort ( 92.6% ), abdominal pain ( 55.6% ), and irregular fever ( 29.6% ).The positive diagnosis rate with US, CT, MRCP, ERCP and EUS was 75.6%, 85.2%, 83.3%, 84.6%, and 88.9%,respectively, and they were helpful for early diagnosis.Among these 27 patients, 19 cases received regional pancreaticoduodenectomy, and 5 cases received pancreaticoduodenectomy in combination with vessel resection,3 cases received bile duct or gallbladder jejunal Roux-en-Y anastomosis, the overall resection rate was 88.9% with no operative mortality, and the post-operation complication rate was 7.4%.The 1,3, and 5 year survival rates were 100%, 70.8% and 41.7%, respectively.Conclusions The periampullary carcinoma in the prejaundice stage has its own clinical characteristics and abnormal image changes.If the diagnosis can be confirmed in the prejaundice stage, it is still an important method to improve the resection rate and prognosis.

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Diagnosis and treatment of 21 blunt pancreatic trauma patients

Zhiyuan JIAN ; Mingyin LAN ; Meng ZHOU ; Yanmei ZHAO

Chinese Journal of Pancreatology.2010;10(6):395-397. doi:10.3760/cma.j.issn.1674-1935.2010.06.005

Objective To investigate the diagnosis and treatment of the blunt pancreatic injury.Methods The clinical data of 21 blunt pancreatic trauma patients admitted to our hospital in the past 5 years were analyzed retrospectively.Results 3 of 21 patients were simple blunt pancreatic injury, while the other 18 patients were complicated with other abdominal organs injuries.According to the grading criteria, the number of Ⅰ , Ⅱ, Ⅲ, Ⅳ, Ⅴ grade injury were 4, 7, 5, 3 and 2 cases, respectively, and only 9 patients were diagnosed by CT before operation, and the diagnosis was confirmed during the operation in 11 patients, and 1 was miediagnosed in the first operation.Single drainage or debridement and drainage were performed in 8 cases and distal pancreatectomy with and without spleen were performed in 5 and 3 cases, respectively, and 3patients received pancreaticojejunostomy, 1 received Whipple operation, the other 1 case underwent duodenorrhahy and diverticulation.The mortality rate was 9.5% (2/21), and the complications of pancreatic pseudocyst and fistula developed in 2 (9.5%, 2/21 ) and 5 (23.8%, 5/21 ) cases respectively, and they were cured by drainage, while the other 12 were cured completely.Conclusions The preoperative misdiagnosis rate of pancreatic trauma is high;CT scan should be used to confirm the diagnosis.Careful pancreatic exploration is mandatory and simple and effective surgical procedures with effective drainage is important for treatment success.

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Kansui root therapy for severe acute pancreatitis with high intra-abdominal pressure

Junming HE ; Shixia CAI ; Xiaosheng ZHONG ; Chengjiang QIU ; Youxing HUANG ; Song WANG ; Xianfeng LIU ; Zhijian TAN ; Bingqin CAI

Chinese Journal of Pancreatology.2010;10(6):392-394. doi:10.3760/cma.j.issn.1674-1935.2010.06.004

Objective To investigate the treatment effects of Kansui root on severe acute pancreatitis (SAP) with abdominal compartment syndrome (ACS).Methods 16 cases of SAP were randomly divided into kansui root treatment group and control group according to random number table.Patients in control group received routine treatment including fasting, anti-shock, antibiotics and nutritional support.And the patients in kansui root group received routine treatment plus kansui root therapy.The clinical and laboratory parameters were determined and compared between the two groups.Results The relieving time of abdominal pain, bowel sound, the recovery time of hyperamylasemia, body temperature and leukocyte count in treatment group was (7.6±2.3)d, (6.1 ±3.1)d, (5.9±3.3)d, (5.2 ±3.2) d, (6.3 ±2.1)d, which were significantly shorter than those in control group [ ( 11.7 ± 2.1 ) d, ( 11.2 ± 2.3d, ( 10.2 ± 2.7) d, (9.2 ± 3.5 ) d, ( 11.1 ±3.3)d, P＜0.01 ) ].At the 3rd, 4th and 5th day, the intra-abdominal pressure in treatment group were also significantly lower than those in control group[ ( 19.8 ±3.1 )cmH2O vs(23.7 ±2.9) cmH2O, ( 12.3 ±2.7) cmH2O vs (21.3±1.5)cmH2O,(8.2±3.1)cmH2O vs (17.3 ±2.3)cmH2O,P＜0.05].Conclusions Severe acute pancreatitis has close relationship with Jiexiong syndrome in traditional chinese medicine.Kansui root is an effective therapy for alleviating high intra-abdominal pressure.

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Dynamic changes and significance of serum IL-1β and sE-selectin in patients with acute pancreatitis

Duanmin HU ; Chunhua ZHOU ; Shaofeng WANG

Chinese Journal of Pancreatology.2010;10(6):386-388. doi:10.3760/cma.j.issn.1674-1935.2010.06.002

Objective To investigate the dynamic changes and relationship between the serum levels of IL-1 β and sE-selectin in acute pancreatitis (AP) patients, and to study the predicative value for severity of AP.Methods Forty one patients with AP were divided into severe acute pancreatitis (SAP, 19 cases) group and mild acute pancreatitis (MAP, 22 cases) group.Their serum were collected at the 1st, 3rd, 7th, and 14th day after admission and the serum concentrations of IL-1 β and sE-selectin were detected by ELISA.Another 20healthy volunteers were selected as controls.Results The serum concentration of IL-1 β in control group was ( 18.71 ±2.43)ng/L, while they were (61.18 ±7.47)ng/L, (33.03 ±5.85)ng/L, (20.73 ±4.07)ng/L in MAP group at the 1st, 3rd, 7th day;and they were (86.91 ± 13.32) rng/L, (81.35 ± 12.71) ng/L,(64.93 ±5.99)ng/L, (21.40±49.13) ng/L at the 1st, 3rd, 7th and 14th day in SAP group.The serum concentration of IL - 1β of patients with SAP were markedly higher than those with MAP and normal controls on the 1st, 3rd, 7th day (P ＜0.05) and they decreased almost to normal on the 14th day.The serum concentration of sE-selectin was ( 10.69 ± 2.51 ) ng/ml, while they were ( 41.60 ± 6.85 ) ng/ml, ( 14.90 ±3.51)ng/ml, (9.85 ±2.88)ng/ml in MAP group at the 1st, 3rd, 7th day;and they were (84.73 ±15.37)ng/ml, ( 95.65 ± 13.06 ) ng/ml, ( 39.41 ± 3.73 ) ng/ml, ( 12.25 ± 2.29 ) ng/mlon the 1st, 3rd, 7 th and 14th day.The serum concentration of sE-selectin of patients with SAP were significantly higher than those with MAP and normal controls on the 1st, 3rd, 7th day (P ＜0.05 ) and there was no significant difference between SAP and control group on the 14th day.There was a positive correlation between the serum level of IL-1 β and sE-selectin in AP on the 1 st day after admission ( r = 0.851, P ＜ 0.01 ).Conclusions The serum concentrations of IL-1 β and sE-selectin are useful for AP severity predication.

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Expression of MCP-1 in the pancreas of piglet with chronic obstructive pancreatitis and its significance

Yang BIAN ; Na TA ; Lin LI ; Qiao ZHENG ; Shunmin ZHANG ; Jianming ZHENG ; Changjing ZUO

Chinese Journal of Pancreatology.2010;10(6):412-414. doi:10.3760/cma.j.issn.1674-1935.2010.06.011

Objective To determine the serum concentration of MCP-1 and the expression of MCP-1 protein in the pancreas in the piglet with chronic obstructive pancreatitis and to explore the role of MCP-1 protein in pancreatic fibrosisits.Methods The piglet model of chronic obstructive pancreatitis was established by incomplete ligation of the pancreatic duct.The piglets were sacrificed at 4, 6, 8 weeks after induction.Pathological changes of pancreas were examined.Pancreatic fibrosis was assessed by VG staining.Serum MCP-1 concentrations were detected by ELISA method.MCP-1 and α-SMA, PDGF, TGF-β1 and NF-κB protein expression were detected by immunohistochemistry.Results The induction was successful in 14 piglets ( 58.3% ).Mild atrophic changes, interstitial fibrosis, chronic inflammatory cell infiltration could be observed in the body and tail of pancreas from the 4th week in the experimental group.The most obvious changes occurred in the 8th week.Stage Ⅰ pancreatic fibrosis occurred in 5 piglets (35.7%), stage Ⅱ in 4 piglets (28.6%), stage Ⅲ in 5 rats ( 35.7% ).Seurm MCP-1 at 4, 6, 8 weeks was ( 102.44 ± 36.25 ) pg/ml,(97.84 ± 28.67) pg/ml, ( 94.32 ± 28.42 ) pg/ml, respectively, and was significantly higher than that in control group [ ( 10.42 ±5.86) pg/ml, (8.58 ±4.86) pg/ml, (8.22 ±4.58) pg/ml, P ＜0.01 ].There was no MCP-1 protein expression in the control group;MCP-1 protein was detected in the successful induction group, and MCP-1 expression was positively correlated with expressions of the PDGF, TGF-β1, α-SMA and NF-κB.Conclusions MCP-1 may play an important role in the course of pancreatic fibrosis in chronic obstructive pancreatitis.

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