OBJECTIVE:To establish an HPLC method for determining the contents of Quercetin and Isorhamnetin in Bupleurum.smithii var.parvifolium Shan et Y.Li.METHODS:The HPLC was performed on Kromasil column C18(250mm? 4.6mm,5? m),using methanol-0.4% phosphoric acid(55∶ 45)as mobile phase,with flow rate at 1.0mL? min-1 and detection wavelength at 256nm.RESULTS:The linear range of Quercetin was 0.08～ 0.40? g(r=0.999 6),with an average recovery rate of 101.02%(RSD=1.53%);that of Isorhamnetin was 0.06～ 0.30? g(r=0.999 2),with an average recovery rate of 101.26%(RSD=2.95%).CONCLUSION:This method is simple and accurate with good reproducibility.It is suitable for the quality control of Bupleurum.smithii var.parvifolium Shan et Y.Li.

Objective:To investigate the role of LncRNA-TUG1 in the injury of intestinal epithelial cells induced by lipopolysaccharide (LPS).Methods:LPS was used to treat HIEC-6 human intestinal epithelial cells for 24 h to construct a sepsis injury model. Whole transcriptome RNA sequencing was used to analyze the expression changes of mRNA, microRNA and lncRNA in HIEC-6 cells after LPS treatment. Real-time fluorescence quantitative (qRT-PCR) and Western blot was performed to detect the expression changes of lncRNA-TUG1, microRNA-132-3p (miR-132-3p), SIRT1 mRNA and SIRT1 protein in HIEC-6 cells after LPS treatment. The expression levels of LncRNA-TUG1, miR-132-3p and SIRT1 were artificially changed by in vitro transfection. qRT-PCR and Western blot were used to confirm the regulatory effect of lncRNA-TUG1 on microRNA-132-3p and SIRT1. CCK-8 and flow cytometry were used to analyze the effects of LncRNA-TUG1, miR-132-3p and SIRT1 on the proliferation and apoptosis of HIEC-6 cells. The dual luciferase report analysis was used to verify the targeting relationship between LncRNA-TUG1, miR-132-3p and SIRT1. Statistical analysis was performed using SPSS 17.0, and differences between the two groups were compared using independent sample t test. Results:RNA sequencing results showed that the expressions of lncRNA-TUG1 and SIRT1 were decreased in HIEC-6 cells after LPS treatment ( t=3.26, P<0.05 and t=2.55, P<0.05), but the expression of miR-132-3p was increased ( t=4.12, P<0.05). In vitro cell experiments, the expression of lncRNA-TUG1 and SIRT1 were decreased in HIEC-6 cells treated with LPS ( t=5.69, P<0.05 and t=5.712, P<0.05), while the expression of miR-132-3p was increased ( t=3.88, P<0.05). Overexpression of lncRNA-TUG1 increased the proliferation rate ( t=6.55, P<0.05) and decreased the apoptosis rate ( t=3.94, P<0.05) of LPS-treated cells. Upregulation of lncRNA-TUG1 decreased the expression of miR-132-3p ( t=4.66, P<0.05), and increased the mRNA and protein levels of SIRT1 ( t=3.91, P<0.05). Transfection of miR-132-3P mimic could inhibit the mRNA ( t=4.08, P<0.05) and protein levels of SIRT1. In LPS-treated cells, the cells co-transfected with miR-132-3pmimic and siRNA-SIRT1 had a lower proliferation rate ( t=4.55, P<0.05 and t=5.67, P<0.05) and a higher apoptosis rate ( t=3.90, P<0.05 and t=4.22, P<0.05) than those transfected with only pcDNA3.1-lncRNA-TUG. Conclusions:lncRNA-TUG1 may act as a ceRNA to regulate miR-132-3p/SIRT1, therefore alleviating HIEC-6 cell injury caused by LPS. Intervention of lncRNA-TUG1/miR-132-3p/SIRT1 regulatory pathway may become a potential strategy to prevent sepsis-induced intestinal mucosal damage.

Pancreatic duct stone is a rare pancreatic disease in clinic, which is often associated with chronic pancreatitis, and could seriously damage the quality of life of patients, and even induce pancreatic cancer. The diagnosis is mainly based on imaging examination, and the treatment methods are diverse. It is necessary to follow the principle of individualized treatment and treat it as soon as possible. This article reviewed the etiology, mechanism, diagnosis, classification and treatment of the disease.

Objective:To study the causes of hemorrhage after laparoscopic pancreaticoduodenectomy (LPD) and to develop countermeasures in its prevention.Methods:The clinical data of 215 patients who underwent LPD at the Department of Hepatobiliary and Pancreatic Surgery of Zhejiang Provincial People's Hospital from December 2013 to May 2020 were reviewed. The patients’ clinical data including gender, age, comorbidities and postoperative complications such as bleeding, pancreatic fistula, biliary fistula and intraperitoneal infection were studied, with the aims to analyze the causes, clinical manifestations and treatment results of post-pancreaticoduodenectomy hemorrhage (PPH) after LPD.Results:Of 215 patients, there were 132 males and 83 females, aged (60.7±10.3) years. PPH occurred in 20 patients, incidence rate was 9.30%(20/215). Early hemorrhage was mainly caused by inadequate hemostasis or loosening of vascular clips, while delayed hemorrhage was mainly caused by gastrointestinal fistula with vascular erosion, arterial injury by intraoperative energy instruments or pseudoaneurysms. Among the 20 patients, 6 patients had early hemorrhage and 14 delayed hemorrhage. There was 1 patient with grade A, 10 with grade B and 9 with grade C hemorrhage. Thirteen patients developed pancreatic fistula, 1 biliary fistula, and 2 intraperitoneal infection. One patient responded well to conservative treatment. Hemostasis was successfully achieved by gastroscopy ( n=1) and interventional therapy ( n=7). Eleven patients required laparotomy for hemostasis. In this study, 14 of 20 patients survivied PPH and 6 patients died. The mortality rate was 30% (6 of 20 patients with PPH). Conclusions:Early hemorrhage was caused by inadequate hemostasis or loosening vascular clips, while delayed hemorrhage was related to gastrointestinal fistula with vascular erosion, arterial injury by intraoperative energy instrument or pseudoaneurysm. Careful hemostasis, adequate protection of blood vessels, and accurate anastomosis should be performed in LPD. DSA angiography should be used for arterial hemorrhage which progressed very rapidly. Interventional therapy including embolism and stenting were means to control arterial bleeding in PPH. Decisive surgical exploration when interventional therapy failed was important in reducing the mortality rate of these patients.

Objective To study the surgical treatment of xanthogranulomatous cholecystitis (XGC).Methods We retrospectively analyzed the clinical data of 56 patients with XGC who underwent surgical treatment at the Zhejiang Provincial People's Hospital from May 2010 to May 2017.Results The diagnosis of XGC was confirmed by histopathology.On preoperative examination of the 56 patients,42 patients had various degrees of increase in the CA19.9 levels,41 patients (73.2％) had thickened gallbladder walls with continuous mucosal linings on ultrasonography,CT,or MRI,and 18 patients (32.1％) had thickening of gallbladder walls with low density nodules.Gallbladder stones were present in 51 patients (91.1％) and 4 patients (7.2％) presented with Mirizzi syndrome.The 41 patients (73.2％) who were diagnosed as XGC before operation under laparoscopic surgery and 7 patients (17.1％) were converted to open surgery.The remaining 15 patients (26.8％) underwent open operation directly because of uncertainty in the diagnosis.All the patients had frozen section during operation.The postoperative pathological results included 21 localizedtype (37.5％) and 35 diffuse type (62.5％) of XGC.All 56 patients had no long-term complications on followed-up for 0.5～ 6 years.Conclusions XGC is a special kind of chronic cholecystitis.There is difficulty in differentiating from gallbladder cancer before surgery.The diagnosis of XGC mainly depends on ultrasonography,CT or MRI.Cholecystectomy is the treatment for XGC.Laparoscopic surgery is the first line treatment for XGC.

Objective To analyze the initial experience of total laparoscopic radical resection for patients with Bismuth type Ⅲa hilar cholangiocarcinoma.Methods A retrospective study was conducted to analyze the clinical data of three patients with Bismuth type Ⅲa hilar cholangiocarcinomatotal who underwent laparoscopic radical resection in Zhejiang Provincial People's Hospital from February to May in 2017.Results The three patients all underwent the operations successfully.The operation time ranged from 490.0 to 580.0 min.The intraoperative blood loss ranged from 300.0 ml to 1 200.0 ml.There was no severe perioperatire complication or death.One patient developed biliary leakage which responded to drainage without reoperation.Another patient developed pleural effusion treated with minimal invasive drainage.The length of postoperative hospital stay ranged from 10.0 to 18.0 days.Histopathology showed two patients with well-differentiated adenocarcinomas and one patient with poorly differentiated adenocarcinoma.The number of lymph nodes harvested ranged from 8 ～ 13.Two patients had no regional lymph node metastasis and one patient had regional lymph node metastasis (1/13).The hilar bile duct resection margins of the three patients were all negative.There was no evidence of tumor recurrence on following up for 7 ～ 10 months.Conclusions It was safe and feasible to carry out total laparoscopic radical resection in selected patients with Bismuth type Ⅲa hilar cholangiocarcinoma.More patients and longer follow-up are required to study the long term oncological results.

Objective To explore the safety and feasibility of laparoscopic hepatectomy combined with radiofrequency ablation for primary liver cancer difficult to manage.Methods A retrospective study was conducted to analyze the clinical data of 16 patients who underwent laparoscopic hepatectomy combined with radiofrequency ablation in the Zhejiang Provincial People's Hospital from Apr 2015 to Dec 2017.Results 2 more tumors were found by intraoperative laparoscopic ultrasound.All patients underwent laparoscopic hepatectomy combined with radiofrequency ablation successfully.There were 2 laparoscopic left hepatectomy combined with radiofrequency ablation,2 laparoscopic right hepatectomy combined with radiofrequency ablation,6 laparoscopic left lateral lobectomy combined with radiofrequency ablation,4 laparoscopic right postrior lobectomy combined with radiofrequency ablation and 2 irregular laparoscopic hepatectomy combined with radiofrequency ablation.The intraoperative blood loss ranged from 100-800 ml.The average operative time was (283 ± 112) min.The length of postoperative hospital stay ranged from 5 to 12 days.The tumor-free survival rate after operation is 100％ (16/16) on 6 to 38 months follow up.Conclusion It is safe and feasible to carry out laparoscopic hepatectomy combined with radiofrequency ablation in difficult to manage primary liver cancer.

Objective To study the safety and feasibility of laparoscopy combined with holmium laser in the treatment of chronic pancreatitis complicated with pancreatic ductal stones.Methods To compare the clinical data in patients who underwent laparoscopy combined with holmium laser (10 patients,group A) with those who underwent laparoscopy only (21 patients,group B) at Zhejiang Provincial People' s Hospital from January 2012 to August 2018.The operation time,intraoperative blood loss,intraoperative conversion rate,pancreatic ductal incision length,postoperative pancreatic fistula rate,length of postoperative hospital stay,residual stone rate and relief of postoperative abdominal pain rate of the two groups were documented and analyzed.Results Three of 31 patients were converted to open surgery.The remaining patients in the two groups were discharged home without any perioperative death.Group A and B were significant differences in the pancreatic ductal incision length (5.0±0.8 vs.6.5±1.0) cm,operation time (289.3±51.6 vs.349.5± 34.7) min,and postoperative hospital stay (8.0± 1.2 vs.10.2± 1.6) d between the two groups (P＜0.05).There were no significant differences in the intraoperative conversion to open rate,intraoperative blood loss,postoperative pancreatic fistula rate,residual stone rate and relief of postoperative abdominal pain rate between the two groups (P ＞ 0.05).Conclusions It was safe and feasible to treat chronic pancreatitis complicated with pancreatolithiasis by laparoscopy.Laparoscopy combined with holmium laser had the added advantages of easy access through the pancreaticojejunostomy,shorter operation time,and less intraoperative blood loss.