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Objective:To compare the efficacies of arthroscopic modified Brostr?m procedure combined with or without peroneal tendon debridement in the treatment of chronic lateral ankle instability (CLAI) concomitant with fibular tendinitis.Methods:A retrospective cohort analysis was conducted on the clinical data of 31 patients with CLAI concomitant with fibular tendinitis, who were treated in Beijing Tongren Hospital, Capital Medical University between March 2019 and December 2021. The patients included 17 males and 14 females, aged 16-57 years [(32.8±9.6)years]. The anterior drawer test and talar tilt test were positive in all patients preoperatively. Diagnosis was confirmed by physical examination and MRI, and calcaneofibular ligament rupture was excluded. Eleven patients received arthroscopic modified Brostr?m procedure combined with peroneal tendon debridement (modified Brostr?m procedure+tendon debridement group), and 20 underwent pure arthroscopic modified Brostr?m procedure (modified Brostr?m procedure group). The operation time, intraoperative blood loss and length of hospital stay were documented. The visual analogue score (VAS) in peroneal tendon area was assessed before operation and at postoperative 2, 6 and 12 weeks. The American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot score and foot and ankle outcome score (FAOS) were assessed before operation and at postoperative 6 and 12 weeks. The anterior drawer test was performed at the last follow-up. The foot and ankle ability measure (FAAM) score was assessed before operation and at the last follow-up. Postoperative wound healing and complications were also observed.Results:All the patients were followed up for 4-19 months [(11.3±3.5)months]. The operation time was (66.0±4.2)minutes in the modified Brostr?m procedure+tendon debridement group, which was significantly longer than (61.5±3.4)minutes in the modified Brostr?m procedure group ( P<0.05). There was no significant difference in intraoperative blood loss or length of hospital stay between the two groups (all P>0.05). Compared with the preoperation, the value of VAS was significantly lowered, and the values of AOFAS ankle-hindfoot score, FAOS and FAAM score were significantly increased at different postoperative timepoints (all P<0.01). No significant differences in the values of VAS, AOFAS ankle-hindfoot score, FAOS or FAAM score were seen between the two groups before operation (all P>0.05). The value of VAS was 3.0(3.0, 4.0) points in the modified Brostr?m procedure+tendon debridement group, being markedly different from 4.0(4.0, 4.0)points in the modified Brostr?m procedure group at 2 weeks postoperatively ( P<0.05). The value of VAS was 2.0(1.0, 3.0)points in the modified Brostr?m procedure+tendon debridement group, being markedly different from 3.0(2.3, 3.0)points in the modified Brostr?m procedure group at 6 weeks postoperatively ( P<0.05). At 12 weeks postoperatively, there was no significant difference in the value of VAS between the two groups ( P>0.05). There were no significant differences in the values of AOFAS ankle-hindfoot score and FAOS between the two groups at 6 or 12 weeks postoperatively (all P>0.05). The anterior drawer test was negative in all patients at the last follow-up. No significant difference was seen in the value of FAAM score between the two groups at the last follow-up ( P>0.05). All incisions were healed well in the first stage after operation, without the occurrence of joint infection, impaired joint motion, nerve injury or deep vein thrombosis. Conclusions:Arthroscopic modified Brostr?m procedure combined with or without peroneal tendon debridement can both improve the foot function in CLAI patients concomitant with fibular tendinitis. However, the combined treatment allows for early pain relief, without increasing the risk of complications, and can therefore contribute to a faster postoperative recovery.
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Objective To investigate the expression of microRNA-1290 in pancreatic cancer and its role in invasion and metastasis of pancreatic cancer.Methods The expression of microRNA-1290 in pancreatic cancer tissue microarray and pancreatic cancer cell lines (AsPC-1,BxPC-3,Capan-2,Panc-1,and MIA PaCa-2) were detected by immunohistochemistry and QT-PCR.The pancreatic cancer cell lines Panc-1 and MIA PaCa-2 in logarithmic growth phase were treated with microRNA-1290 inhibitor,and the invasion and metastasis ability of pancreatic cancer cells were detected by Transwell and wound healing asssay.Western Blot was used to detect the expression of invasion and metastasis-associated proteins cyclooxygenase 2 (COX-2) and matrix metalloproteinase 2(MMP-2) in pancreatic cancer cell lines.Results (1) The expression of microRNA-1290 in pancreatic cancer tissues was significantly higher than that in normal pancreatic tissues and adjacent tissues (P ＜ 0.05).(2) Compared with pancreatic normal epithelial cells (HPDE),the expression of microRNA-1290 was significantly higher in different pancreatic cancer cell lines (P ＜ 0.05).The expression level of MicroRNA-1290 in Panc-1 and MIAPaCa-2 pancreatic cancer cells was significantly higher than that in other pancreatic cancer cell lines (P ＜ 0.05).(3) The number of invasive and metastatic cells was significantly decreased after treatment with microRNA-1290 inhibitor (P ＜0.05).(4) The expression of MMP-2 and COX-2 were decreased in Panc-1 and MIAPaCa-2 pancreatic cancer cells treated with MicroRNA-1290 inhibitor.Conclusion The expression of MMP-2 and COX-2 may be involved in the invasion and metastasis of pancreatic cancer cell by regulating the expression of microRNA-1290 in pancreatic cancer.
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Objective To investigate the therapeutic effects of endoscopic retrograde cholangiopancreatography (ERCP) and laparoscopic bile duct exploration lithotomy (LBDEL) in treatment of intra/extra-hepatic duct stones. Methods There were 110 patients whose intrahepatic stones located in Ⅰ , Ⅱ hepatic duct and 378 patients whose stones only located in the common bile duct. These patients respectively underwent LBDE combined with choledochoscope laser lithotripsy or ERCP combined with endoscopic sphincterotomy (EST) and endoscopic nasobiliary drainage (ENBD) to remove the stones. Common bile ducts were performed primary suture or T tube placement in the LBDEL cases. The evaluation was carried out for perioperative complications and postoperative recovery of the surgical methods. Results The residual stone rate was 31.82% in 110 cases. The rate was higher in ERCP group (51.06%) than that in LBDEL group (17.46%) (P < 0.05). Postoperative recovery was better in LBDEL group than that in ERCP group. The residual stone rate was 8.20% in 378 cases. The rate was lower in ERCP group (3.68%) than that in LBDEL group(11.63%) (P < 0.05). Between the two groups, there had no statistical significance in postoperative recovery. The incidences of bile leakage and pulmonary infection were higher in LBDEL group than in ERCP group. The incidences of abdominal cavity infection, acute pancreatitis, digestive tract perforation and gastrointestinal bleeding were higher in ERCP group than that in LBDEL group. 2 of the 378 patients occurred death were happened digestive tract perforation which were induced during ERCP procedure. Conclusion LBDEL and ERCP demonstrated the same therapeutic effects in the treatment of common bile duct stones. However, ERCP has no large advantages in the treatment of hepatolithiasis, and shows higher complication rates. LBDEL has a significant curative effect for intra-and extrahepatic bile duct calculi and can maintain the integrity of Oddi sphincter. This technology is easy to spread to the basic-level hospital to benefit the majority of patients.
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Objective To investigate the effect and mechanism of high concentration glucose on cholesterol absorption of human colon cancer epithelial Caco-2 cells.Methods The experimental study was used.(1) CCK-8 detected cell proliferation:the proliferation rate changes of Caco-2 cells were detected by CCK-8 when different concentrations (12.5,100.0,300.0,700.0,1 000.0,1 388.0 mmol/L) of glucose solution effects on Caco-2 cells in order to ensure the half hindering concentration of glucose concentration on Caco-2 cells.(2)Cholesterol absorption of Caco-2 cells was detected:Caco-2 cells were divided into the cholesterol group,cholesterol plus ezetimibe (cholesterol inhibitor) group and blank control group.Cholesterol group:100 μmol/L cholesterol solution and different concentrations (5.0,25.0,50.0 mmol/L) of glucose solution were added.Cholesterol plus ezetimibe group:100 μmol/L ezetimibe,100 μmol/L cholesterol solution and different concentrations (5.0,25.0,50.0 mmol/L) of glucose solution were added.Blank control group:DMEM culture medium and corresponding concentrations of DMSO were added.The cholesterol absorption amounts of Caco-2 cells were measured.(3) The relative expressions of ATP binding cassette G8 (ABCG8),ATP binding cassette G5 (ABCG5),Nickman-Pick CI Like 1 (NPC1L1) and scavenger receptor class B type Ⅰ (SR-B Ⅰ) were examined by Western blot in the different groups.Caco cells were divided into the glucose group,glucose plus ezetimibe group and control group.The different concentrations (5.0,25.0,50.0 mmol/L) of glucose solution were added into the glucose group,different concentrations (5.0,25.0,50.0 mmol/L) of glucose solution and 100 μmol/L ezetimibe were added into the glucose plus ezetimibe group,and 100 μmol/L ezetimibe were added into the control group.The relative expressions of ABCG8,ABCG5,NPC1L1 and SR-B Ⅰ were detected by Western blot.Measurement data were presented as (x) ±s,repeated measure variance analysis was used to perform variation trend test,and t test was utilized to conduct comparisons among groups.Results (1) CCK-8 results showed:proliferation rates of Caco-2 cells with the glucose solution concentration of 12.5,100.0,300.0,700.0,1 000.0 and 1 388.0 mmol/L were 1.380 ±0.043,1.238 ±0.072,0.736 ±0.035,0.336 ±0.021,0.316 ±0.020 and 0.288 ±0.010,respectively,with a statistically significant difference in the proliferation rates (F =11.019,P ＜ 0.05).The half hindering concentration of glucose solution on Caco-2 cells was 283.54 mmol/L.(2)Cholesterol absorption of Caco-2 cells:① the cholesterol absorption amounts of Caco-2 cells with the glucose solution concentration of 5.0,25.0 and 50.0 mmol/L were 0.282 ± 0.042,0.380 ± 0.063,0.390 ± 0.060 in the cholesterol group and 0.042 ± 0.012,0.197 ± 0.015,0.277 ± 0.029 in the cholesterol plus ezetimibe group,respectively,showing a statistically significant difference between the 2 groups (F =55.566,P ＜ 0.05).②There was a statistically significant difference in cholesterol absorption amounts of Caco-2 cells with different glucose solution concentration in the cholesterol group (F =79.117,P ＜ 0.05).The cholesterol absorption amounts of Caco-2 cells with the glucose solution concentrations of 5.0 mmol/L was lower than that with the glucose solution concentrations of 25.0 mmol/L and 50.0 mmol/L,respectively (t =11.207,11.532,P ＜0.05).There was no statistically significant difference in the cholesterol absorption amounts of Caco-2 cells between the glucose solution concentrations of 25.0 mmol/L and 50.0 mmol/L (t =12.389,P ＞ 0.05).③There were statistically significant differences in cholesterol absorption amounts of Caco-2 cells with the glucose concentration of 5.0 mmol/L and 25.0 mmol/L between cholesterol group and cholesterol plus ezetimibe group (t =10.908,10.644,P ＜ 0.05).(3) The results of Western blot showed:① the relative expression of NPC1L1 protein in Caco-2 cells with the glucose solution concentrations of 5.0,25.0 and 50.0 mmol/L were respectively 0.277 ±0.019,0.558 ±0.015,0.576 ±0.003 in the glucose group and 0.057 ±0.002,0.054 ±0.005,0.077 ±0.005 in the glucose plus ezetimibe group,showing a statistically significant difference (F =482.207,P ＜0.05).② The relative expression of NPC1L1 protein of Caco-2 cells with the different concentration of glucose solution in the glucose group were compared,with a statistically significant difference (F =8.112,P ＜ 0.05).There was a statistically significant difference in the relative expression of NPC1L1 protein in Caco-2 cells with the different concentration of glucose solution in the glucose plus ezetimibe group (F =11.708,P ＜ 0.05).③ The relative expression of NPC1L1 protein in Caco-2 cells with the glucose solution concentrations of 5.0,25.0 and 50.0 mmol/L in the glucose group was statistically different from that in the glucose plus ezetimibe group (t =8.112,11.708,13.920,P ＜ 0.05).Conclusion High concentration glucose solution could promote the reabsorption of cholesterol through increasing NPC1L1 protein expression in Caco-2 cells,and increase the risk of suffering from cholelithiasis in diabetes patients.
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Objective To observe the effect of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on cholesterol gallstones formation in C57BL/6 mice with diet-induced cholesterol gallstone,and then explore the potential mechanism.Methods Fifty C57BL/6 mice were randomly divided into 5 groups (10 mice in each group),referring to control group,experimental group,experimental plus DHA group,experimental plus EPA group,as well as experimental plus DHA and EPA group.The mice in control group were fed with regular diet,and the rest of the mice with lithogenic diet (LD).Subsequent to feeding the mice with separate diets for two weeks,EPA and/or DHA (70 mg · kg-1 · d-1) were orally administered for eight weeks,while the LD feeding was continued during this period.After a total of 10 weeks,the mice were dissected to observe the gallstone formation.The levels of serum lipids,total cholesterol (TC) and phospholipids (PL) in bile,and TC in the liver were tested,and the protein expression of HMGCR,SRBI,ABCG5/ABCG8,CYP7A1 and ABCB11genes in the liver of mice was measured.Results Compared with the experimental group,the experimental plus EPA group had significantly lower TC in liver (0.033 ±0.008 mmolo/g) and bile (1.807 ±0.381 mmolo/L),and lower relative protein expression levels of HMGCR (0.545±0.098),ABCG5 (0.418±0.089) and ABCG8 (0.501 ±0.151)in liver (P＜ 0.05).The contents of TC in liver and bile,and the protein expression of HMGCR,ABCG5andABCG8 in liver were 0.048 ± 0.006 mmol/g and 2.662 ± 0.339 mmolo/L,and 1.011 ± 0.213,1.037 ± 0.276 and 1.266 ±0.312,respectively.No significant differences were observed between experimental plus DHA group and experimental group (P ＞ 0.05).Conclusions EPA could prevent the cholesterol gallstone formation in mice by decreasing the expression of HMGCR and ABCG5/8 genes in liver,therefore reducing cholesterol synthesis and blocking cholesterol transport from liver to bile as well as diminishing cholesterol content in the bile.However,the inhibition effect of DHA on cholesterol gallstone formation was not obvious.
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Objective To investigate the effects of miR-100 on the proliferation of MIA PaCa-2 and CFPAC-1 cells through targeting fibroblast growth factor receptor 3 (FGFR3).Methods miR-100 expression levels in 17 cancer tissues and 17 nonmalignant tissues were examined by Real-time PCR.The effect of miR-100 overexpression on cell proliferation was examined by CCK-8 assay in vitro.Luciferase assay was used to confirm that miR-100 could directly target FGFR3.Real-time PCR and Western blot were used to examine the expression of FGFR3 in miR-100 overexpressing pancreatic cancer cells.The predicted target gene of miR-100,FGFR3,was downregulated by siRNA,and its effect on cell proliferation was also examined.Cell proliferation was analyzed using CCK-8 and Edu assay.Results miR-100 was lowly expressed in pancreatic cancer tissues (P ＜ 0.05).In pancreatic cancer cells,the transfection of lv-miR-100 was able to upregulate the endogenous expression of miR-100 and inhibit the cell proliferation (P ＜0.05).Luciferase assay showed FGFR3 was the direct target of miR-1O0.FGFR3 was significantly downregulated by overexpressing miR-100 in pancreatic cancer cells (P ＜0.05),and FGFR3 knockdown by specific siRNA exerted the similar effect as miR-100 overexpression (P ＜ 0.05).Conclusions Our study identified a new miRNA regulator,miR-100,and clarified a novel mechanism of how miR-100 regulates cell proliferation in pancreatic cancer.The strategy of overexpressing the tumor suppressor miR-100 may provide a new therapeutic approach for treating patients with pancreatic cancer.
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Objective To investigate the effects and mechanism of expression of microRNA-100 in the tissues of pancreatic cancer and its relationship with clinicopathological factor.Methods The clinical data of 17 patients with pancreatic cancer who were admitted to the Affiliated Hospital of Guizhou Medical University between January 2013 and March 2014 were retrospectively analyzed.The surgical specimens were collected for study.The expression of microRNA-100 in the pancreatic tissues were detected by real-time fluorescent quantitative polymerase chain reaction (RT-PCR) and its relationship with clinicopathological factors was analyzed.The MIA PaCa-2 cells and CFPAC-1 cells were infected by lv-microRNA-100.MIA PaCa-2 cells were divided into the M group (microRNA-100 were infected) and N group (empty vectors were infected),and CFPAC-1 cells were divided into the C group (microRNA-100 were infected) and D group (empty vectors were infected).The expressions of microRNA-100 in the CFPAC-1 cells and MIA PaCa-2 cells were detected by RT-PCR and cell cycles were detected by flow cytometry.The expressions of Cyclin D1 protein in the CFPAC-1 cells and MIA PaCa-2 cells were detected by Western blot.Measurement data with normal distribution were presented as (x) ± s and comparison among groups were done by t test.Results The results of RT-PCR showed that the relative quantitative expression of microRNA-100 in the pancreatic cancer tissues was 0.046 ± 0.020,which was significantly different from 0.097 ± 0.017 in the adjacent tissues (t =2.789,P ＜ 0.05).There were significant differences between the tumor differentiation degree and tumor staging in patients with pancreatic cancer (t =2.563,2.135,P ＜0.05).The results of RT-PCR showed that the relative quantitative expression of microRNA-100 in the M group was 0.097 ±0.012,which was significantly higher than 0.018 ± 0.006 in the N group (t =4.410,P ＜ 0.05).The relative quantitative expression of microRNA-100 in the C group was 0.084 ± 0.021,which was significantly higher than 0.023 ± 0.010 in the D group (t =5.351,P ＜ 0.05).The results of flow cytometry showed that the percentage of cells between G0-G1 were 45.3％ ±0.7％ in the M group and 30.6％ ±0.7％ in the N group,with a significant difference (t =5.564,P ＜ 0.05).The percentage of cells between G0-G1 were 58.8％ ± 1.0％ in the C group and 42.6％ ± 0.7％ in the D group,with a significant difference (t =7.771,P ＜ 0.05).The results of Western blot showed that the relative quantitative expression of Cyclin D1 protein were 0.352 ± 0.081 in the M group and 0.872 ± 0.134 in the N group,with a significant difference (t =7.651,P ＜ 0.05).The relative quantitative expression of Cyclin D1 protein was 0.410 ± 0.121 in the C group,which was significantly different from 0.979 ± 0.232 in the D group (t =8.712,P ＜ 0.05).Conclusion Low expression of microRNA-100 in pancreatic cancer tissues may be correlated with tumor differentiation degree and tumor staging,it can inhibit tumor cells proliferation by reducing expression of Cyclin D1 protein.
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Objective:To investigate the expression of Plk1 ( Polo-like kinase 1 ) and Chk1/2 ( Checkpoint kinase 1/2 ) in primary hepatic carcinoma tissue and HepG2 cell. Methods: Using immunohistochemistry chemical method detected expression of Plk1,Chk1/2 protein in 40 cases of primary hepatic carcinoma tissue and 16 cases of non-tumor tissue of liver. Western blot was applied to detect the expression of Plk1 and Chk1/2 protein in HepG2 cells, and gray value was measured by using the quantitative analysis. Results:The positive rate of Plk1,Chk1/2 protein expression in primary hepatic carcinoma was 57. 5%,75. 0% and 22. 5%respectively,compared with positive rate in the liver of non-tumor tissue were 0%,25. 0% and 56. 3%. The expression of Plk1 and Chk1 protein in primary hepatic carcinoma tissue is higher than that in non-tumor tissue of liver,and the difference was statistically sig-nificant( PChk1>Chk2. Conclusion: Plk1,Chk1 protein in primary hepatic carcinoma was up-regulated,while Chk2 protein was down-regulated in these tissues. The expression degree was Plk1> Chk1>Chk2. There were relatively selective expression in primary hepatic carcinoma tissue of Plk1,Chk1 protein,then Plk1 and Chk1 might be ideal targets for therapy of primary hepatic carcinoma.
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PurposeTraumatic pancreatitis which has a high mortality rate is likely to be misdiagnosed. This study aims to analyze the clinical manifestations and CT findings of traumatic pancreatitis, so as to improve its early diagnosis and treatment.Materials and Methods The clinical manifestations and CT images of 25 patients with traumatic pancreatitis confirmed by operation or post-treatment review were analyzed retrospectively. Pancreatic injuries were classified as superficial lesions (with the depth of trauma less than 50% of the thickness of pancreas) and deep lesions (with the depth of trauma more than 50% of the thickness of pancreas). The clinical manifestations, CT findings and the complicated organ injuries in these two types of pancreatic trauma were analyzed.Results Eight patients had superficial lesions, and 17 patients were with deep lesions. Nine patients had complicated organ injuries. Patients with deep lesions showed a more severe abdominal pain, nausea, vomiting, rebound tenderness and muscular tension than those patients with superficial lesions. The serum amylases increased in all the patients. Pancreatic-relevant complications including pancreas pseudocyst, pancreatic fluid leakage and peritonitis occurred in 7 patients who accepted a delayed operation. Three out of 8 patients with superficial pancreatic injuries were missed on plain CT scan in the first time. Among 17 patients with deep pancreatic trauma, 12 had incomplete laceration, 5 had complete laceration, and 1 was missed in the first time. The direct CT features of pancreatic trauma were focal abnormal attenuation and/or discontinuity in pancreatic parenchyma.Conclusion The clinical manifestations of patients with traumatic pancreatitis are complicated. The direct CT features of pancreatic trauma include heterogeneous density of pancreatic parenchyma and/or interruption. Trauma's depth is closely related to the main injury of pancreatic duct. It is worth to be aware of the indirect signs such as peripancreatic oozy and other viscera damages.
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Objective To investigate transcription intermediary factor 1β (TIF1β) expression in paracancerous pancreatic tissue and pancreatic tumor tissue by using tissue microarray.The relationship between TIF1β expression and clinicopathological factors in patients with pancreatic cancer was discussed.Method Tissuc microarray and immunohistochemical assay were utilized to detect expression of TIF1β protein in pancreatic cancer tissues and the corresponding non-tumor tissues from 91 cases.Results TIF1β protein were present in pancreatic cancer tissues as well as corresponding non-tumor tissues with varying degrees of expression,and was located in the nucleus.TIF1β expression in pancreatic cancer tissue was significantly higher than that of adjacent tissues (P ＜ 0.05).And it was noted that there was close correlations between TIF1β expression and clinical pathological staging,lymph node metastasis and TNM grading (P ＜ 0.05).Conclusions TIF1β is highly expressed in pancreatic cancer,clinically correlated with pathological staging,lymph node metastasis and TNM grading.TIF1β may play an important role in development of pancreatic cancer.
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Objective To investigate the clinical efficacy of two types of duodenum-preserving pancreatic head resection (Beger procedure and Berne procedure) for chronic pancreatitis with mass in the head of the pancreas.Methods The clinical data of 46 patients with chronic pancreatitis and mass in the head of the pancreas who were admitted to the Affiliated Hospital of Guiyang Medical College from September 2008 to April 2012 were retrospectively analyzed.There were 24 patients received Beger procedure (Beger group),and 22 received Beme procedure (Berne group).The complications,life quality and pain after the operation were evaluated.Patients were followed up via phone call and out-patient examination till April 2013.The measurement data were analyzed using the Mann-Whitney U test,and the constituent ratios were compared using the chi-square test.Results The operation time and volume of blood loss were (377 ± 21) minutes and (746 ± 129) mL in the Beger group,and (323 ± 17) minutes and (577 ± 111)mL in the Berne group,with significant difference between the 2 groups (U=14.0,88.0,P ＜0.05).Four patients in the Beger group and 1 in the Berne group were complicated with pancreatic leakage,with no significant difference between the 2 groups (x2=0.714,P ＞ 0.05).The scores of life quality evaluation (physical condition,work capacity,cognitive ability,emotion,social competence and overall life quality) were 82 ± 14,74±24,90 ± 18,78±20,83 ± 18,73 ± 18 in the Beger group,and 79 ± 16,71 ±20,92 ±21,76 ± 18,80 ±21,70 ± 16 in the Berne group,with no significant difference between the 2 groups (U =177.5,183.5,187.5,178.0,189.5,192.0,P ＞ 0.05).The scores of symptom evaluation (fatigue,nausea and vomitting,pain,anorexia,dyspnea,sleep disorders,obstipation,diarrhea,financial worries) were 28 ± 16,24 ± 10,20±12,23 ± 14,4 ± 1,32 ± 12,6 ±2,18 ± 14,36± 18 in the Beger group,and 26 ± 18,26 ±20,22 ± 16,26 ± 16,3 ± 1,30 ± 10,5 ± 1,16 ± 12,38 ± 20 in the Berne group,with no significant difference between the 2 groups (U=194.5,215.5,182.5,180.5,213.0,199.0,195.0,184.5,181.5,P＞0.05).In the Beget group,19 patients did not have acute onset of pain,and 5 patients had acute onset of pain once a year; 6 patients were administered antalgesic occasionally.In the Berne group,20 patients did not have acute onset of pain,and 2 patients had acute onset of pain once a year; 4 patients were administered antalgesic occasionally,with no significant difference between the 2 groups (x2=0.485,0.041,P ＞ 0.05).All the patients were followed up,and the median time of follow-up was 36.3 months.No perforation of duodenum and steatorrhea was observed.No patient died perioperatively.Conclusion The clinical efficacy of the Berne procedure is similar to that of the Beger procedure,while the Berne procedure has advantages of easy manipulation and less operation time.
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Objective To evaluate the MUC1 promoter's role in driving gene expression in pancreatic cancer and its therapeutic significance.Methods Two plasmids were made.The plasmid pEGFP-MUC1N1 contained MUC1 promoter fragment connected to the pEGFP-N1 vector with the EGFG reporter gene.The pShuttle-MUC1-EGFP plasmid contained MUC1 promoter fragment and EGFP reporter gene connected to pShuttle plasmid.Lipofectamine 2000 was used to transfect the two plasmids into cells of MUC1-positive human pancreatic cell line Panc-1 and MUC1-negative human cervical carcinoma Hela.Fluorescence microscopy and flow cytometry compared the specificity and activity of the MUC1 promoter and CMV promoter.Results Reporter gene EGFP-positive cells 48 hours after transfection with pEGFP-MUC1-N1 and pShuttleMUC1-EGFP plasmid were 69.6％ and 63.6％ respectively,in Panc-1 cells,and 4.2％ and 3.7％ respectively,in Hela cells.Conclusions MUC1 promoter can drive reporter gene activity in MUC1-positive tumor cells targeting functional expression.There is potentially a use of targeted therapy in pancreatic cancer at the genetic level.
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Objective To investigate whether KAP-1 could induce CSC-self renewal in pancreatic cancer cell lines.Methods KAP-1 expression was examined in 14 cases of pancreatic cancer with immunohistochemistry.KAP-1 shRNA amplified by PCR was inserted into pGC-LV in vector,and then identified by restriction endonuclease digestion and nucleotide sequencing.The lentiviral vector pGC-shRNA-KAP-1 was co-transfected with pHelper 1.0 and pHelper 2.0 packaging plasmids into HEK 293T cells,and the lentivirus was collected and virus titer was measured.The expressions of KAP-1 and vimentin were detected by Western blot and RT-qPCR when human pancreatic cancer cell line Panc-1 was infected by the lentivirus.Sphere forming assay was conducted to assess the capacity of CSC or CSC-like cell self-renewal in this study.Results The KAP-1 expression level in cancerous tissues on immunohistochemistry was significantly higher than in the corresponding normal tissues (P=0.002).After infected by lentivirus,the expressions of KAP-1 and vimentin were knocked down,which could be detected by Wesren blot and RT-qPCR.Compared to Panc-1-GFP (NC) cells,the outcomes suggested that knocking down the expression of KAP-1 could decrease the formation of pancreatospheres,which further suggested the capacity of CSC-self-renewal in primary and secondary pancreatospheres of Panc-1-shRNA-KAP-1 and NC cells.Conclusions KAP-1 expression in pancreatic cancer tissues has been identified.The lentiviral vector for shRNAs targeting KAP-1 was constructed successfully.The formation of pancreatospheres decreased by knocking down the KAP 1 gene.KAP-1 is involved in the regulation of CSC phenotype.The mechanism is probably related to the upregulated expression of the EMT marker vimentin.
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Objective To assess the clinical significance of QT interval dispersion (QTd) in the diagnosis and prognosis of early heart damage in patients with severe acute pancreatitis.Methods All patients received complete ECG,There were 58 patients with SAP (the SAP group) and 189 patients with mild acute pancreatitis (the control group).These patients were analyzed retrospectively and 60 normal people were used as the healthy control group.The QT interval were measured respectively in serial 12-lead electrocardiogram and QTd,QTcd were calculated.Result QTd and QTcd were significantly longer in the SAP group than in the MAP group and in the healthy control group (P＜0.01).QTd,QTcd were not remarkably extended in the MAP group than in the healthy control group (P＞0.05).Conclusions QTd and QTcd have clinical values to diagnose and to predict early heart damage in patients with SAP.They might be useful in evaluating the condition of cardiac function in patients with SAP.
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Objective To explore the overexpression of the 14-3-3 sigma (14-3-3σ) gene effects proliferation in pancreatic cancer cells.Methods The 14-3-3σ protein level in 46 pancreatic cancers and 9 normal pancreases specimens were analyzed by immunohistochemistry.The mRNA and protein expression of 14-3-3σ in pancreatic carcinoma cell lines (BxPC3,CAPAN-1,PANC-1,AsPC-1,SW1990,MiaPaCa-2 and CFPAC-1) were detected by RT-qPCR and Western blot respectively.A recombinant plasmid of pEGFP-14-3-3σ was constructed and transfected into the pancreatic cancer cell PANC-1 by liposome,and the expression of 14-3-3σ was detected by Western blot and real time fluorescence quantitative PCR.Cell proliferation activity was determined by MTS assay.Results The 14-3-3σ protein level was higher in pancreatic cancer tissue than in normal pancreatic tissue.mRNA and protein expression of 14-3-3σ was highest in BxPC3,high in AsPC-1,SW1990 and CFPAC-1,low in PANC-1 and Capan-1,and lowest in MiaPaCa-2.The successfully constructed pEGFP-14-3-3σ was confirmed by RT-qPCR and Western blot.MTS assay showed cell proliferation activity was significantly enhanced by overexpression of the 14-3-3σ gene compared to negative and blank control cells.Conclusion The expression of 14-3-3σ was higher in pancreatic cancer compared with normal pancre atic tissue,and the 14-3-3σ gene enhanced the cell proliferation activity of PANC-1.Therefore,14-3-3σ may play an important role in pancreatic cancer development.
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Objective To investigate the possibility of transfection of adipose-derived mesenchymal stem cells (ADMSCs) into the insulin-secreting cells in vitro,and assay the secretion of insulin of ADMSCs in high and low glucose environment.Methods The ADMSCs that untransfected were in the control group,the ADMSCs that contained PcDNA3.1 were in the vacant vector group,and the ADMSCs that contained PcDNA3.1-hINS were in the recombinant vector group.After transfection,the recombinant vector group were sub-divided into the 1,6,12,18 days groups.According to the concentrations of glucose,the recombinant vector 18 days group were divided into the high glucose group and low glucose group.Human insulin gene was amplified by RT-PCR,and the eukaryotic expression recombinant vector PcDNA3.1-hINS that contained the human insulin gene was constructed.The ADMSCs were obtained by digestion and centrifugation,and then underwent flow cytometry for identification.The transcription of insulin DNA was assayed by RT-PCR,and the levels of insulin were assayed by ELISA.Glucose test was done in the recombinant vector 18 days group.The measurement data was shown in the format of (x) ± s,the measurement data in multiple groups were compared by randomized analysis of variance,and the comparison among groups was performed by the t test.ResuIts The expressions of CD44,CD90,CD106 were positive,and the expressions of CD34,CD45 and CD11b were negative.No insulin DNA transcription was detected in the control group and vacant vector group.The levels of insulin secreted were (4.7 ± 0.8) mIU/L,(8.8 ± 0.5) mIU/L,(8.9 ± 0.8)mIU/L,(8.6 ± 0.6)mIU/L in the recombinant vector 1,6,12,18 days group,which were significantly higher than (1.3 ± 0.6) mIU/L in the control group and (1.7 ± 0.8) mIU/L in the vacant vector group (t =10.09,32.64,22.20,55.53 ; 9.23,27.56,19.43,51.25,P ＜ 0.05).There were significant differences in the levels of insulin secreted between the recombinant vector 1 day group and the recombinant vector 6,12,18 days groups (t =12.77,12.26,13.93,P ＜0.05).There were no significant difference in the levels of insulin secreted between the recombinant vector 6,12,18 days groups (F =45.67,P ＞ 0.05).There was a significant difference in the level of insulin secreted between the high glucose group and the low glucose group (t =2.03,P ＜ 0.05).The result of the glucose stimulation test was negative.Conclusion The ADMSCs are transfected into insulinsecreting cells in vitro successfully,and the secretion of insulin is stable.Although the secretion of insulin can't change in line with the concentration of glucose,it is a new seed cell for the treatment of diabetes with stem cells.
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Objective To investigate the effects of KAP-1 in promoting the epithelial-mesenchymal transition (EMT) of pancreatic cancer cell line Capan-2.Methods The lentiviral vector of LV-plenti-GFP-KAP-1 was constructed.Capan-2 cells were divided into the experimental group (cells transfected by lentiviral vector of LVplenti-GFP-KAP-1),negative control group (cells transfected by empty vector) and blank control group 1 (cells cultured in 1640 medium plus 10％ fetal calf serum).Capan-2 cells in the experimental group were subdivided into the miR-100-5p inhibitor transfection group (cells transfected with miR-100-5p inhibitor),empty vector control group (cells transfected with microRNAs inhibitor),blank control group 2 (cells cultured in 1640 medium plus 10％ fetal calf serum).The lentivirus was identified by double endonuclease restriction and sequencing,and the virus titer was detected.The morphological changes of the cells were observed after transfecting lentiviral vector of LV-plenti-GFP-KAP-1 to the Capan-2 cells.The expressions of KAP-1,genes of EMT proteins and mRNA of miR-100-5p were detected by real-time quantitative polymerase chain reaction.The protein expressions of KAP-1,EMT proteins in all the groups were detected by Western blot.The measurement data were presented by mean ± standard deviation,and were analyzed using the analysis of variance.Results The lentiviral vector of LV-plentiGFP-KAP-1 was successfully constructed,and the virus titer was 2 × 108 TU/ml.Compared with the control group,the mesenchymal transition of the Capan-2 cells was detected in the experimental group after transfecting the Capan-2 cells with lentiviral vector of LV-plenti-GFP-KAP-1 for 48 hours.The relative mRNA expressions of KAP-1,N-cadherin,vimentin,E-cadherin,miR-100-5p were 1.77 ± 0.83,2.62 ± 0.71,2.50 ± 0.21,7.20 ± 1.17 and 1.81 ±0.40 in the experimental group,5.03 ±0.29,5.07 ±1.53,3.83 ±0.57,7.83 ±0.78,7.01 ± 0.96 in the negative control group,5.13 ± 1.14,5.81 ± 1.49,4.92 ± 0.90,3.07 ± 0.36,6.87 ± 0.35 in the blank control group 1,with significant difference among the 3 groups (F =5.99,7.62,7.88,6.62,4.64,P ＜0.05).The relative mRNA expression of KAP-1 in the miR-100-5p inhibitor transfection group,empty vector control group,blank control group 2 were 1.56 ± 0.42,4.89 ± 0.61,5.20 ± 0.38,with significant difference among the 3 groups (F =5.14,P ＜ 0.05).The relative mRNA expressions of vimentin in the miR-100-5p inhibitor transfection group,empty vector control group,blank control group 2 were 3.10 ± 1.37,3.44 ± 0.94,3.08 ±1.16,with no significant difference among the 3 groups (F =0.49,P ＞ 0.05).The results of western blot showed that the relative protein expressions of KAP-1,N-cadherin,vimentin,E-cadherin were 2.77 ± 1.99,1.31 ±0.38,4.25 ± 0.63,0.62 ± 0.06 in the experimental group,0.83 ± 0.46,0.41 ± 0.37,1.03 ± 0.33,1.17 ± 0.45 in the negative control group,0.71 ± 0.26,0.08 ± 0.04,1.37 ± 0.92,3.04 ± 0.65 in the blank control group 1,with significant difference among the 3 groups (F =5.54,4.68,3.19,8.18,P ＜ 0.05).The relative protein expression of KAP-1 in the miR-100-5p inhibitor transfection group,empty vector control group,blank control group 2 were 2.27 ±0.71,0.56 ±0.43,0.61 ±0.39,with significant difference among the 3 groups (F =4.81,P ＜0.05).The relative protein expressions of vimentin in the miR-100-5p inhibitor transfection group,empty vector control group,blank control group 2 were 3.19 ± 0.55,3.93 ± 0.06,3.61 ± 0.73,with no significant difference among the 3 groups (F =0.04,P ＞ 0.05).Conclusion KAP-1 promotes the EMT of Capan-2 cells by specifically down-regulating the miR-100-5p expression.
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Comprehensive treatment dominated by surgery is the mainstay in the treatment of hepatic cancer,and hepatectomy is still the most effective treatment method.Bile duct reconstruction after hepatectomy is still the difficult point for the treatment of hepatic cancer complicated by bile duct invasion.A 45-year-old patient with hepatic cancer and gallstone was admitted to the Affiliated Hospital of Guiyang Medical College,magnetic resonance cholangiopancreatography and enhanced computed tomography indicated that intrahepatic duct was dilated and tumor had invaded both left and right hepatic ducts.Cholecystectomy,mesohepatectomy,duct to duct anastomosis of left hepatic duct and common hepatic duct,duct to duct anastomosis of right hepatic duct and cystic duct were performed during the operation.The patient was cured 2 weeks after surgery.
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Objective To explore the impact of insulin secretory function of adult islet cells from barium-alginate microencapsulation in vitro. Methods After weighting the pancreas,human pancreatic islets were isolated with type V collagenase and purified by Ficoll's discontinuous density gradient centrifugation.The islet cell yield and purity were evaluated with microscope by DTZ staining.Human islets were coated by barium-alginate microencapsulation,and the viability was assessed by insulin release assay in vitro.Results After isolation,the average number of islet was about 3600 ±447 per gram pancreas.It was about 2140±207 after purification with more than 70％ purity.At day 2,4 and 6 after islet cell culture in vitro,basal insulin concentrations from the culture medium was measured,and the mean insulin concentration (mU/L) in media of the microencapsulated islet group at 2nd,4th and 6th day were 3.302±1.63、3.504±1.10 and 2.921±1.13 respectively,and those non-microencapsulated islet group were 3.814 ± 1.49、4.175 ±1.60、3.617± 1.34.There were no significant differences between these two groups (P＞0.05).Conclusions The bioartificial pancreas has effective insulin secretory function in vitro without being affected by barium-alginate microencapsulation.
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Objective To investigate the methods and feasibility of islets isolation and purification,and to get more islets with high purity and good function for clinical transplantation.Methods After being weighted,human pancreatic islets were isolated with type Ⅴ collagenase,and purified by Ficoll's discontinuous density gradient centrifugation.The yield and purity of islets were evaluated by dithizone(DTZ) staining under microscope,and the viability was assessed by insulin release assay in vitro.Results The average number of islets was about 3 600 ± 447 per gram pancreas after isolation and it was about 2140 ±207 after purification with more than 70％ purity.2,4,6 days after islet cell culture,the basal insulin concentration of the culture medium was measured,and it was 3.302 ± 1.63,3.504 ±1.10,and 2.921 ±1.13 (mIU/L/100 islets) respectively.Conclusion Collagenase digest and Ficoll's discontinuous density gradient centrifugation are effective methods for isolation and purification of human pancreatic islets.