Regulatory effect of miR-210 in hypoxia-induced epithelial-mesenchymal transition of pancreatic cancer PANC1 cells / 中华胰腺病杂志

Objective:To explore the regulatory role of miR-210 in hypoxia-induced epithelial-mesenchymal transition (EMT) of pancreatic cancer PANC1 cells.Methods:PANC1 cells cultured in normoxia and hypoxia were established in normoxia group and hypoxia group. Recombinant plasmid carrrying miR-210 mimics and miR-210 antagomirs were constructed. The recombinant plasmids were transfected with PANC1 cells cultured in normoxia and hypoxia by liposome method to establish cell lines of miR-210 overexpression (miR-210 mimics normoxia group) and miR-210 expression inhibition (miR-210 antagomirs hypoxia group). The blank plasmids were transfected to establish blank plasmid normoxia group and blank plasmid hypoxia group. Relative expression levels of miR-210 for PANC1 cells were determined by qRT-PCR in each group. Western blot was used to measure the expressions of HIF-1α, NF-κB and EMT related protein such as E-cadherin, β-catenin, vimentin and N-cadherin. Cell relative viability under gemcitabine and in vitro cell invasion ability were detected by CCK8 and Transwell chamber experiments, respectively. Results:The relative expressions of miR-210 in hypoxia group and miR-210 mimics normoxia group were significantly higher than those in normoxia group and blank plasmid normoxia group. However, there were significantly lower in miR-210 antagomirs hypoxia group than those in blank plasmid hypoxia group. The expression levels of HIF-1α, NF-κB and mesenchymal cell markers such as vimentin and N-cadherin in hypoxia group were significantly higher than those in normoxia group (0.74±0.06 vs 0.40±0.05, 1.58±0.16 vs 1.09±0.13, 0.46±0.04 vs 0.17±0.02, 1.27±0.07 vs 0.40±0.03) and the epithelial cell markers such as E-cadherin and β-catenin were significantly lower (0.40±0.07 vs 0.77±0.10, 0.35±0.02 vs 0.94±0.08). The expression levels of HIF-1α, NF-κB, vimentin and N-cadherin in miR-210 mimics normoxia group were significantly higher than those in blank plasmid normoxia group (0.91±0.08 vs 0.40±0.06, 1.52±0.17 vs 1.05±0.14, 0.82±0.06 vs 0.66±0.07, 0.76±0.04 vs 0.46±0.03) and E-cadherin and β-catenin were significantly lower (0.38±0.07 vs 0.65±0.09, 0.50±0.03 vs 0.94±0.08). The expression levels of HIF-1α, NF-κB, vimentin and N-cadherin in miR-210 antagomirs hypoxia group were significantly lower than those in blank plasmid hypoxia group (0.31±0.05 vs 0.55±0.06, 0.68±0.05 vs 1.11±0.13, 0.41±0.03 vs 0.74±0.07, 0.69±0.06 vs 0.78±0.05), while E-cadherin and β-catenin were significantly higher (0.72±0.13 vs 0.50±0.07, 0.71±0.04 vs 0.54±0.05). All the differences among the groups were statistically significant (all P value <0.05). Under gemcitabine, the relative viability of PANC1 cells in hypoxia group and miR-210 mimics normoxia group were significantly higher than those in normoxia group and blank plasmid normoxia group at 48 h (1.10±0.10 vs 0.76±0.05, 1.46±0.11 vs 1.12±0.09) and 72 h (1.12±0.13 vs 0.76±0.05, 1.54±0.13 vs 1.12±0.09) accordingly. However, there were significantly lower in miR-210 antagomirs hypoxia group than those in blank plasmid hypoxia group at 48 and 72 h (0.75±0.09 vs 1.10±0.10, 1.19±0.12 vs 1.46±0.11). All the differences among the groups were statistically significant (all P value <0.05). The number of transmembrane cells in hypoxia group and miR-210 mimics normoxia group was significantly higher than those in normoxia group and blank plasmid normoxia group, respectively (417.50±81.22 vs 228.30±47.71, 371.30±72.81 vs 245.00±33.62 per high field), while those in miR-210 antagomirs hypoxia group was significantly lower than those in blank plasmid hypoxia group (228.30±54.01 vs 433.30±65.63 per high field). All the differences among the groups were statistically significant (all P value <0.05). Conclusions:miR-210 could weaken the sensitivity to gemcitabine and promote the invasion of PANC1 cells by regulating the occurrence of the hypoxia-induced epithelial-mesenchymal transition.

Splenic vascular flow control in the use of laparoscopic spleen-preserving distal pancreatectomy (Kimura) / 中华普通外科杂志

Objective:To investigate the techniques used in blood flow control of Kimura laparoscopic spleen-preserving pancreatectomy (LSPDP).Methods:Forty·five patients with benign or low-grade malignant pancreatic diseases undergoing LSPDP at Huzhou Central Hospital from May 2014 to Oct 2021 were analyzed retrospectively. Patients were divided into splenic vascular flow control group ( n=22) and routine management group ( n=23). Results:There was no significant difference in gender, age, BMI, accompanying symptoms, hypertension, diabetes, lesion size and pathological diagnosis between the two groups (all P>0.05). A higher overall spleen preservation rate (90.9% vs. 52.2%, χ2=8.213, P=0.004), lower incidence of morbidity with Clavien grade ≥ Ⅱ (22.7% vs. 73.9%, χ2=9.911, P=0.002) and shorter postoperative hospital stay [(9.6±4.5) d vs. (14.3±6.6) d, t=2.447, P=0.008] were achieved in the vascular flow control group compared with those in the routine group. Conclusion:Splenic vascular flow control techniques improve the success rate of spleen preservation in laparoscopic distal pancreatectomy, reduce the postoperative complications and shorten the postoperative hospital stay.

Laparoscopic cholecystectomy in patients with a history of gastrectomy / 中华普通外科杂志

Objective To investigate the risk factors of laparoscopic cholecystectomy (LC) in patients with previous gastrectomy.Methods This retrospective study reviewed a database of patients who underwent LC at our hospital during the period of Jan 2010 and May 2015.Results Comparesd with those of no history of gastrectomy the average operation time in patients with previous gastrectomy was longer (t =15.608,P ＜ 0.05) the intraoperative blood loss was higher (t =3.061,P ＜ 0.05),and the operation time of patients with a long interval (＞ 5 years) between gastrectomy and LC was shorter (t =6.405,P ＜ 0.01).The conversion rate did not differ between the two groups (P ＞ 0.05),but the conversion rate significantly reduced after a comprehensive preoperative evaluation procedure (x2 =15.282,P ＜ 0.01).Conclusion LC for benign gallbladder diseases is safe,effective and feasible in patients with a history of gastrectomy,if a comprehensive preoperative evaluation is adopted and in experinced hands.

Laparoscopic partial splenectomy for benign splenic neoplasms / 中华普通外科杂志

Objective To evaluate laparoscopic partial splenectomy (LPS) for benign splenic neoplasms.Methods From Sep 2016 to Aug 2017,7 patents with splenic benign lesions underwent laparoscopic partial splenectomy,results were compared with 15 cases receiving total splenectomy.Results All the patients underwent laparoscopic total or partial splenectomy successfully without conversion.There was no statistic difference in operative time,intra-operative blood loss and transfusion between the two groups.However,the morbidity of LPS group was significant lower (x2 =17.679,P =0.000),and no thrombocytosis occurred in LPS patients.Additionally,the patients received LPS recovered quicker and the postoperative hospital stay was shorter (5.7 ± 2.3) d vs.(15.3 ± 3.4) d (t =6.741,P =0.000 1) with a lower medical expenditure (19 288 ± 2 760) Yuan vs.(27 737 ± 4 626) Yuan (t =4.442,P =0.000 3).No thrombocytosis and recurrence were observed during follow up.Conclusions LPS is safe and effective to cure spleen benign lesions with quicker recovery and lower expenditure.

The impact of antiviral therapy on postoperative intrahepatic recurrence of hepatitis B related hepatocellular carcinoma / 中华肝胆外科杂志

Objective To study the impact of antiviral therapy on patients with postoperative intrahepatic recurrence of hepatitis B related hepatocellular carcinoma (HCC).Methods According to the inclusion and exclusion criteria,65 patients with HCC who were managed in our hospital from January 2004 to December 2011 for intrahepatic recurrence and had indications for antiviral treatment were included into this study.The patients were divided into two groups ; the antiviral and the control groups.Relevant data between these two groups such as cumulative survival after recurrence,Child-Pugh grade,HBV-DNA,HBeAg,AFP at the time of recurrence and 6 months later were studied.An analysis on multiple-factors was carried for survival after recurrence at 2 years.Results When compared with the control group,the antiviral group had better cumulative survival in all the cases and in the TACE cases (P ＜ 0.05 respectively).The cumulative survival in the RFA cases was not significantly different between the two groups (P ＞ 0.05).A comparison was carried out in patients who received antiviral therapy which was combined with one or more other therapies (TACE,RFA,reoperation):(a) antiviral therapy only (8 patients) ; (b) combined with one therapy (22 patients) ; (c) combined with two therapies (10 patients) ; (d) combined with three therapies (2 patients).All P values of a:b,a:c,a:d,b:c,b:d were less than 0.05.The blood HBV-DNA of the two groups was significantly different at the time 6 months after recurrence (P ＜ 0.05).The results of multivariate analysis showed the 2-year survival was significantly correlated with recurrent tumor size,primary tumor class,antiviral therapy or not after recurrence,presence of absence of cirrhosis.All P values were less than 0.05.Conclusions Antiviral therapy had remarkable clinical impact on HCC patients with postoperative intrahepatic recurrence and with indication for antiviral treatment.Patients had better prognosis if antiviral therapy was combined with one or more other therapies.

Effects of timing of enteral nutrition in the severe acute pancreatitis / 中华消化外科杂志

Objective To explore the effects of timing of enteral nutrition (EN) in the severe acute pancreatitis (SAP).Methods The rabbit model of SAP was constructed,and 40 experimental rabbits were divided randomly into the experimental group (20 rabbits) and the control group(20 rabbits).(1) The parenteral nutrition (PN) was administered to rabbits in the experimental group,and then EN was administered after bowel sound restoration.(2) EN was administered to the rabbits in the control group at hour 12 after PN.The values of TNF-α,IL-6 and IL-8 on post-modeling day 1,3 and 7 and intra-abdominal pressures at hour 1 before EN and on post-modeling day 1 and 3 were respectively examined.The experimental animals were sacrificed on post-modeling day 7 and 10.The expression of nuclear factor-kappa B (NF-κB) was detected by immunohistochemistry and bacteria from mesenteric lymph nodes were cultured.The measurement data with normal distribution were presented as (x) ± s.The trend comparison between groups and pairwise comparison were analyzed using the repeated measures ANOVA and t test.Results The 40 rabbit models of SAP were constructed successfully.(1) The results of serum test showed that the values of TNF-α,IL-6 and IL-8 in the experimental group on post-modeling day 1,3and 7 were (6.9 ±2.1)ng/L,(5.9 ± 1.8) ng/L,(5.3 ± 1.8) ng/L and (109 ± 17) ng/L,(96 ± 16) ng/L,(83 ± 17)ng/L and (89 ±20)ng/L,(78 ±21)ng/L,(70 ± 19)ng/L,respectively.The values of TNF-α,IL-6 and IL-8 in the control group on post-modeling day 1,3 and 7 were (7.4 ± 1.5) ng/L,(7.1 ± 1.6) ng/L (6.8 ±1.7) ng/L and (101 ± 19) ng/L,(98 ± 18) ng/L,(93 ±20) ng/L and (91 ± 14) ng/L,(91 ± 16) ng/L,(83 ±19)ng/L,respectively.There were significant differences in the changing trends of TNF-α and IL-6 between the 2 groups (F =7.947,19.386,P ＜ 0.05),with no significant difference in the changing trend of IL-8 between the 2 groups (F =2.756,P ＞ 0.05).(2)The intra-abdominal pressures in the experimental group at hour 1 before EEN and on post-modeling day 1 and 3 were (3.5 ± 1.6) cmH2O (1 cmH2O =0.098 kPa),(4.3 ± 1.7) cmH2Oand (3.6 ± 1.8) cmH2O,which were significantly different from (4.9 ± 1.9) cmH2O,(5.7 ± 2.2) cmH2O and (4.5 ± 1.6)cmH2O in the control group (F =7.042,P ＜0.05).(3) The expression of NF-κB in the tissues of pancreas was localized mainly in the cell nucleus.The expression of NF-κB in the experimental group on postmodeling day 7 was 5.0 ± 2.7,which was significantly different from 7.0 ± 2.9 in the control group (t =2.236,P ＜ 0.05).(4) The amount of bacteria culture from mesenteric lymph nodes in the experimental group on postmodeling day 7 and 10 were (4.7 ±0.9) × 103 cfu/g and (4.1 ±0.7) × 103 cfu/g,which were significantly different from (5.5 ± 1.0) × 103 cfu/g and (4.6 ± 0.7) × 103 cfu/g in the control group (t =2.382,2.126,P ＜0.05).Conclusion The selective timing of EN can effectively improve the recovery of rabbits with SAP,and the efficacy of EN administering based on the recovery of bowel function may be better than immediate EN administering.

Clinical application value of laparoscopic intraoperative cholangiography in laparoscopic cholecystectomy / 中国基层医药

Objective To investigate the clinical application value of the laparoscopic intraoperative cholan-giography in laparoscopic cholecystectomy,and summarize the experience.Methods The clinical data of 169 cases of laparoscopic cholecystectomy intraoperative cholangiography were analyzed retrospectively.Results 169 patients were successfully completed,131 cases recovered well and no complications occurred after operation.29 patients showed hyperamylasemia,of which 3 patients had intractable hyperamylasemia,8 patients complicated with secondary acute pancreatitis,with fasting,gastrointestinal decompression,enzyme inhibition(plus somatostatin)and acid,analgesic, anti infection,rehydration treatment after remission.Conclusion Laparoscopic cholecystectomy intraoperative cholan-giography is a safe and reliable diagnostic technique,on the occurrence of biliary residual stones in prevention of post-operative prevention and timely detection of bile duct injury during operation and improves the success rate of repair of bile duct injury has important clinical value;control adaptation of intraoperative cholangiography has important clinical significance of reasonable application license.

Therapeutic effect of Shenmai injection on acute pancreatitis and mechanism / 中华胰腺病杂志

Objective To observe the therapeutic effect of Shenmai injection on rats with acute edematous pancreatitis (AEP) and its mechanism.Methods The model of AEP in rats was induced by intraperitoneal injection of caerulein every one hour for 7 times.One hour after last injection,intravenous Shenmai injection (5 ml · kg-1 · d-1) was given to rats in Shenmai injection treatment (SI) group,while same amount of normal saline was given to rats in control (AEP) group.One hour and 1,3,5,7 d after AEP induction,the blood was taken,and the method of double antibody sandwich enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of amylase,platelet activating factor (PAF),vascular endothelial growth factor (VEGF).The rats were sacrificed at 7 and 14 d,and pancreatic tissue samples were harvested.RT-PCR and Western Blot were used to determine the expression of NF-κB mRNA and protein,and normal pancreatic tissue samples were used as control.The positive expression of CD31 in pancreatic tissue was determined by immunohistochemical method to calculate microvascular density (MVD).Results The serum levels of amylase and PAF in both groups were increased and gradually decreased 3 days later,while the decrement in SI group was greater than that of AEP group.At the 5th day after AEP induction,the serum levels of amylase in SI group and AEP group were (4569 ± 158),(5056 ± 198) U/L,and the serum levels of PAF were (25.30 ± 4.76),(36.06 ± 5.57)μg/L,and the difference between the two groups was statistically significant (P ＜ 0.05).The serum levels of VEGF were increased 1 d after AEP induction,and the increase in SI group was greater than that of AEP group 3 d later,and the serum levels of VEGF were (139.78 ±24.30),(118.51 ±21.70)pg/ml at 5th day,and the difference between the two groups was statistically significant (P ＜ 0.05).The expression levels of NF-κB mRNA and protein of pancreatic tissue in SI group were O.834 ±0.031 and 0.49±0.24,and MVD was 6.41 ± 1.14,while the corresponding values in AEP group were 1.000 ± 0.059,0.93 ± 0.45,3.62 ± 0.89,and the difference between the two groups was statistically significant (P ＜ 0.05 or P ＜ 0.01).Conclusions In the course of acute pancreatitis,Shenmai injection has the therapeutic effects of promoting new angiogenesis,improving the microcirculation,reducing the inflammatory cascade.