Primary cultures of pancreatic stellate cells(PSCs) remain an important basis for in vitro study.However,effective methods for isolating abundant PSCs are currently lacking.We report on a novel approach to isolating P...Primary cultures of pancreatic stellate cells(PSCs) remain an important basis for in vitro study.However,effective methods for isolating abundant PSCs are currently lacking.We report on a novel approach to isolating PSCs from normal rat pancreases and human pancreatic ductal adenocarcinoma(PDAC) tissue.After anaesthesia and laparotomy of the rat,a blunt cannula was inserted into the pancreatic duct through the anti-mesentery side of the duodenum,and the pancreas was slowly infused with an enzyme solution until all lobules were fully dispersed.The pancreas was then pre-incubated,finely minced and incubated to procure a cell suspension.PSCs were obtained after the cell suspension was filtered,washed and subject to gradient centrifugation with Nycodenz solution.Fresh human PDAC tissue was finely minced into 1×1×l mm^3 cubes with sharp blades.Tissue blocks were placed at the bottom of a culture plate with fresh plasma(EDTA-anti-coagulated plasma from the same patient,mixed with CaCL) sprinkled around the sample.After culture for 5-10 days under appropriate conditions,activated PSCs were harvested.An intraductal perfusion of an enzyme solution simplified the procedure of isolation of rat PSCs,as compared with the multiple injections technique,and a modified outgrowth method significantly shortened the outgrowth time of the activated cells.Our modification in PSC isolation methods significantly increased the isolation efficiency and shortened the culture period,thus facilitating future PSC-related research.展开更多
Acute pancreatitis(AP)can vary widely in its severity,from being clinically self-limiting to a rapidly fatal course.[1]Necrotizing pancreatitis(NP)is the most serious form and is associated with a poor prognosis;the m...Acute pancreatitis(AP)can vary widely in its severity,from being clinically self-limiting to a rapidly fatal course.[1]Necrotizing pancreatitis(NP)is the most serious form and is associated with a poor prognosis;the mortality rate is approximately 15%,or up to 30%for cases of infected necrotizing pancreatitis(INP),which often progresses to sepsis and multiple organ failure,the major cause of death and severe complications.[2]The approach to the management of INP has significantly changed during the last 20 years and continues to evolve with the accumulation of experience,new techniques,and research data.Major surgical intervention and debridement were once the mainstay of therapy for patients with symptomatic necrotic foci,but a minimally invasive approach that focuses on percutaneous and/or endoscopic drainage or debridement is now favored.[2]The"step-up"approach,which involves minimally invasive techniques,represents a new paradigm for the treatment of patients with INP,and open pancreatic debridement is now considered to be the final step in the treatment of NP.[2]展开更多
Background:Pancreaticoduodenectomy(PD)has been widely applied in general hospitals in China;however,there is still a lack of unified standards for each surgical technique and procedure.This survey is intended to inves...Background:Pancreaticoduodenectomy(PD)has been widely applied in general hospitals in China;however,there is still a lack of unified standards for each surgical technique and procedure.This survey is intended to investigate the current status of digestive tract reconstruction after PD in university hospitals in China.Method:A cross-sectional survey was conducted among the members of the Young Elite Pancreatic Surgery Club of China by using the Questionnaire for Digestive Tract Reconstruction after Pancreaticoduodenectomy.The questionnaire was disseminated and collected by point-to-point communication via WeChat public platforms.Results:A total of 73 valid questionnaires were returned from 65 university hospitals in 28 provincial divisions of China's Mainland.The respondents who performed PD surgery with an annual volume of over 100 cases accounted for 63%.Generally,laparoscopic PD was performed less often than open PD.Child and Whipple reconstructions accounted for 70%and 26%,respectively.The sequence of pancreatoenteric,biliary-enteric,and gastrointestinal reconstruction accounted for 84%of cases.In pancreatoenteric anastomosis,double-layer anastomosis is the most commonly employed type,accounting for approximately 67%,while single-layer anastomosis accounts for 30%.Of the double-layer anastomoses,duct-to-mucosa/dunking(94%/4%)pancreatojejunostomy was performed with duct-mucosa using the Blumgart method(39%)and Cattel-Warren(29%),with continuous/interrupted sutures in the inner layer(69%/31%)and continuous/interrupted sutures in the outer layer(53%/23%).In single-layer anastomosis,continuous/interrupted sutures accounted for 41%/45%.In hepatojejunostomy,single-layer/double-layer suture accounted for 79%/4%,and continuous/interrupted suture accounted for 75%/9%.Forty-six percent of the responding units had not applied double-layer biliary-intestinal anastomosis in the last 3 years,75%of the responding surgeons chose the anastomosis method according to bile duct diameter,with absorbable/non-absorbable suture accounting for 86%/12%.PD/pylorus-preserving PD accounted for 79%/11%of gastrojejunostomy(GJ)cases,the distance between GJ and hepaticojejunostomy<30,30-50,and>50 cm were 11%,75%,and 14%,respectively.Antecolic/retrocolic GJ accounted for 71%/23%of cases.Twenty-two percent of GJ cases employed Braun anastomosis,while 55%and 19%of GJ cases used linear cutting staplers/tube-type staplers,respectively;60%/14%were reinforced/not reinforced via manual suturing after stapler anastomosis.Manual anastomosis in GJ surgery employed absorbable/non-absorbable sutures(91%/9%).Significant differences in reconstruction techniques were detected between different volumes of PD procedures(<100/year and>100/year),regions with different economic development levels,and between north and south China.Conclusion:Digestive tract reconstruction following PD exists heterogeneity in Chinese university hospitals.Corresponding prospective clinical studies are needed to determine the consensus on pancreatic surgery that meets the clinical reality in China.展开更多
Background Pancreaticoduodenectomy(PD)has been widely applied in general hospitals in China;however,there is still a lack of unified standards for each surgical technique and procedure.This survey is intended to inves...Background Pancreaticoduodenectomy(PD)has been widely applied in general hospitals in China;however,there is still a lack of unified standards for each surgical technique and procedure.This survey is intended to investigate the current status of digestive tract reconstruction after PD in university hospitals in China.Method A cross-sectional survey was conducted among the members of the Young Elite Pancreatic Surgery Club of China by using the Questionnaire for Digestive Tract Reconstruction after Pancreaticoduodenectomy.The questionnaire was disseminated and collected by point-to-point communication via WeChat public platforms.Results A total of 73 valid questionnaires were returned from 65 university hospitals in 28 provincial divisions of China's Mainland.The respondents who performed PD surgery with an annual volume of over 100 cases accounted for 63%.Generally,laparoscopic PD was performed less often than open PD.Child and Whipple reconstructions accounted for 70%and 26%,respectively.The sequence of pancreatoenteric,biliary-enteric,and gastrointestinal reconstruction accounted for 84%of cases.In pancreatoenteric anastomosis,double-layer anastomosis is the most commonly employed type,accounting for approximately 67%,while single-layer anastomosis accounts for 30%.Of the double-layer anastomoses,duct-to-mucosa/dunking(94%/4%)PJ was performed with duct-mucosa using the Blumgart method(39%)and Cattel-Warren(29%),with continuous/interrupted sutures in the inner layer(69%/31%)and continuous/interrupted sutures in the outer layer(53%/23%).In single-layer anastomosis,continuous/interrupted sutures accounted for 41%/45%.In hepatojejunostomy,single-layer/double-layer suture accounted for 79%/4%,and continuous/interrupted suture accounted for 75%/9%.Forty-six percent of the responding units had not applied double-layer biliary-intestinal anastomosis in the last 3 years,75%of the responding surgeons chose the anastomosis method according to bile duct diameter,with absorbable/non-absorbable suture accounting for 86%/12%.PD/pylorus-preserving PD accounted for 79%/11%of GJ cases,the distance between GJ and HJ<30 cm,30-50 cm and>50 cm were 11%,75%,and 14%,respectively.Antecolic/retrocolic GJ accounted for 71%/23%of cases.Twenty-two percent of GJ cases employed Braun anastomosis,while 55%and 19%of GJ cases used linear cutting staplers/tube-type staplers,respectively;60%/14%were reinforced/not reinforced via manual suturing after stapler anastomosis.Manual anastomosis in GJ surgery employed absorbable/non-absorbable sutures(91%/9%).Significant differences in reconstruction techniques were detected between different volumes of PD procedures(<100/year and>100/year),regions with different economic development levels,and between north and south China.Conclusion Digestive tract reconstruction following PD exists heterogeneity in Chinese university hospitals.Corresponding prospective clinical studies are needed to determine the consensus on pancreatic surgery that meets the clinical reality in China.展开更多
基金partially supported by the National Natural Science Foundation of China(81300351, 81272239,81170336)the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD,JX10231801)
文摘Primary cultures of pancreatic stellate cells(PSCs) remain an important basis for in vitro study.However,effective methods for isolating abundant PSCs are currently lacking.We report on a novel approach to isolating PSCs from normal rat pancreases and human pancreatic ductal adenocarcinoma(PDAC) tissue.After anaesthesia and laparotomy of the rat,a blunt cannula was inserted into the pancreatic duct through the anti-mesentery side of the duodenum,and the pancreas was slowly infused with an enzyme solution until all lobules were fully dispersed.The pancreas was then pre-incubated,finely minced and incubated to procure a cell suspension.PSCs were obtained after the cell suspension was filtered,washed and subject to gradient centrifugation with Nycodenz solution.Fresh human PDAC tissue was finely minced into 1×1×l mm^3 cubes with sharp blades.Tissue blocks were placed at the bottom of a culture plate with fresh plasma(EDTA-anti-coagulated plasma from the same patient,mixed with CaCL) sprinkled around the sample.After culture for 5-10 days under appropriate conditions,activated PSCs were harvested.An intraductal perfusion of an enzyme solution simplified the procedure of isolation of rat PSCs,as compared with the multiple injections technique,and a modified outgrowth method significantly shortened the outgrowth time of the activated cells.Our modification in PSC isolation methods significantly increased the isolation efficiency and shortened the culture period,thus facilitating future PSC-related research.
基金This work was supported by a grant from the National Natural Sciences Foundation of China(No.81672449)。
文摘Acute pancreatitis(AP)can vary widely in its severity,from being clinically self-limiting to a rapidly fatal course.[1]Necrotizing pancreatitis(NP)is the most serious form and is associated with a poor prognosis;the mortality rate is approximately 15%,or up to 30%for cases of infected necrotizing pancreatitis(INP),which often progresses to sepsis and multiple organ failure,the major cause of death and severe complications.[2]The approach to the management of INP has significantly changed during the last 20 years and continues to evolve with the accumulation of experience,new techniques,and research data.Major surgical intervention and debridement were once the mainstay of therapy for patients with symptomatic necrotic foci,but a minimally invasive approach that focuses on percutaneous and/or endoscopic drainage or debridement is now favored.[2]The"step-up"approach,which involves minimally invasive techniques,represents a new paradigm for the treatment of patients with INP,and open pancreatic debridement is now considered to be the final step in the treatment of NP.[2]
文摘Background:Pancreaticoduodenectomy(PD)has been widely applied in general hospitals in China;however,there is still a lack of unified standards for each surgical technique and procedure.This survey is intended to investigate the current status of digestive tract reconstruction after PD in university hospitals in China.Method:A cross-sectional survey was conducted among the members of the Young Elite Pancreatic Surgery Club of China by using the Questionnaire for Digestive Tract Reconstruction after Pancreaticoduodenectomy.The questionnaire was disseminated and collected by point-to-point communication via WeChat public platforms.Results:A total of 73 valid questionnaires were returned from 65 university hospitals in 28 provincial divisions of China's Mainland.The respondents who performed PD surgery with an annual volume of over 100 cases accounted for 63%.Generally,laparoscopic PD was performed less often than open PD.Child and Whipple reconstructions accounted for 70%and 26%,respectively.The sequence of pancreatoenteric,biliary-enteric,and gastrointestinal reconstruction accounted for 84%of cases.In pancreatoenteric anastomosis,double-layer anastomosis is the most commonly employed type,accounting for approximately 67%,while single-layer anastomosis accounts for 30%.Of the double-layer anastomoses,duct-to-mucosa/dunking(94%/4%)pancreatojejunostomy was performed with duct-mucosa using the Blumgart method(39%)and Cattel-Warren(29%),with continuous/interrupted sutures in the inner layer(69%/31%)and continuous/interrupted sutures in the outer layer(53%/23%).In single-layer anastomosis,continuous/interrupted sutures accounted for 41%/45%.In hepatojejunostomy,single-layer/double-layer suture accounted for 79%/4%,and continuous/interrupted suture accounted for 75%/9%.Forty-six percent of the responding units had not applied double-layer biliary-intestinal anastomosis in the last 3 years,75%of the responding surgeons chose the anastomosis method according to bile duct diameter,with absorbable/non-absorbable suture accounting for 86%/12%.PD/pylorus-preserving PD accounted for 79%/11%of gastrojejunostomy(GJ)cases,the distance between GJ and hepaticojejunostomy<30,30-50,and>50 cm were 11%,75%,and 14%,respectively.Antecolic/retrocolic GJ accounted for 71%/23%of cases.Twenty-two percent of GJ cases employed Braun anastomosis,while 55%and 19%of GJ cases used linear cutting staplers/tube-type staplers,respectively;60%/14%were reinforced/not reinforced via manual suturing after stapler anastomosis.Manual anastomosis in GJ surgery employed absorbable/non-absorbable sutures(91%/9%).Significant differences in reconstruction techniques were detected between different volumes of PD procedures(<100/year and>100/year),regions with different economic development levels,and between north and south China.Conclusion:Digestive tract reconstruction following PD exists heterogeneity in Chinese university hospitals.Corresponding prospective clinical studies are needed to determine the consensus on pancreatic surgery that meets the clinical reality in China.
文摘Background Pancreaticoduodenectomy(PD)has been widely applied in general hospitals in China;however,there is still a lack of unified standards for each surgical technique and procedure.This survey is intended to investigate the current status of digestive tract reconstruction after PD in university hospitals in China.Method A cross-sectional survey was conducted among the members of the Young Elite Pancreatic Surgery Club of China by using the Questionnaire for Digestive Tract Reconstruction after Pancreaticoduodenectomy.The questionnaire was disseminated and collected by point-to-point communication via WeChat public platforms.Results A total of 73 valid questionnaires were returned from 65 university hospitals in 28 provincial divisions of China's Mainland.The respondents who performed PD surgery with an annual volume of over 100 cases accounted for 63%.Generally,laparoscopic PD was performed less often than open PD.Child and Whipple reconstructions accounted for 70%and 26%,respectively.The sequence of pancreatoenteric,biliary-enteric,and gastrointestinal reconstruction accounted for 84%of cases.In pancreatoenteric anastomosis,double-layer anastomosis is the most commonly employed type,accounting for approximately 67%,while single-layer anastomosis accounts for 30%.Of the double-layer anastomoses,duct-to-mucosa/dunking(94%/4%)PJ was performed with duct-mucosa using the Blumgart method(39%)and Cattel-Warren(29%),with continuous/interrupted sutures in the inner layer(69%/31%)and continuous/interrupted sutures in the outer layer(53%/23%).In single-layer anastomosis,continuous/interrupted sutures accounted for 41%/45%.In hepatojejunostomy,single-layer/double-layer suture accounted for 79%/4%,and continuous/interrupted suture accounted for 75%/9%.Forty-six percent of the responding units had not applied double-layer biliary-intestinal anastomosis in the last 3 years,75%of the responding surgeons chose the anastomosis method according to bile duct diameter,with absorbable/non-absorbable suture accounting for 86%/12%.PD/pylorus-preserving PD accounted for 79%/11%of GJ cases,the distance between GJ and HJ<30 cm,30-50 cm and>50 cm were 11%,75%,and 14%,respectively.Antecolic/retrocolic GJ accounted for 71%/23%of cases.Twenty-two percent of GJ cases employed Braun anastomosis,while 55%and 19%of GJ cases used linear cutting staplers/tube-type staplers,respectively;60%/14%were reinforced/not reinforced via manual suturing after stapler anastomosis.Manual anastomosis in GJ surgery employed absorbable/non-absorbable sutures(91%/9%).Significant differences in reconstruction techniques were detected between different volumes of PD procedures(<100/year and>100/year),regions with different economic development levels,and between north and south China.Conclusion Digestive tract reconstruction following PD exists heterogeneity in Chinese university hospitals.Corresponding prospective clinical studies are needed to determine the consensus on pancreatic surgery that meets the clinical reality in China.
