Liver cirrhosis can lead to continuous pathological elevation of portal venous pressure, resulting in various complications such as esophageal and gastric varices, ascites, hepatic encephalopathy, and secondary circulatory dysfunction. Multimodality therapy is the main treatment method for portal hypertension, among which surgical treatment is mainly used to control and prevent massive esophagogastric variceal bleeding. Currently, interventional therapy and liver transplantation have gradually become a routine method for surgical intervention and can significantly improve patients’ quality of life, but devascularization and shunt are still the mainstream surgical procedures. Each surgical strategy for cirrhotic portal hypertension has its own characteristics. It will be a trend in the future to evaluate patients’ general conditions and clinical features and select individualized treatment regimens to achieve the best treatment outcome.

OBJECTIVE: To study the protective effect of isoflurane preconditioning on hepatic ischemia-reperfusion (I/R) injury mediated by the noncanonical pyroptosis pathway. METHODS: Thirty C57BL/6 mice were randomly divided into sham-operated group, isoflurane group and I/R group, and in the latter two groups, hepatic I/R injury was induced by clamping the portal vein for 30 min. In isoflurane group, the mice were pretreated with 1.4% isoflurane 30 min before the surgery. The protective effect of isoflurane preconditioning against hepatic I/R injury was evaluated by assessing the pathological score of HE staining of the liver tissue and serum ALT and AST levels. Serum IL-1β and IL-18 levels and the protein expression of GSDMS were detected by ELISA and Western blotting to evaluate the inhibitory effect of isoflurane preconditioning on pyroptosis. Western blotting and immunofluroescence were used to detect the protein expression of caspase-11 in the liver tissues to evaluate the inhibitory effect of isoflurane preconditioning on noncanonical pyroptosis pathway. RESULTS: The Suzuki's score of the liver tissue was significantly higher in I/R group than in the sham group ( < 0.05), while the score in the isoflurane group was significantly lower than that in the I/R group ( < 0.05). Serum ALT and AST levels significantly increased in the sham group ( < 0.05), and were significantly lower in isoflurane group than in I/R group ( < 0.05). The serum levels of IL-1β and IL-18 were significantly higher in I/R group than in sham group ( < 0.05), and were significantly lower in isoflurane group than in I/R group ( < 0.05). The expression of GSDMD in the I/R group was significantly higher than that in sham group, and was significantly lower in isoflurane group than in I/R group ( < 0.05). The hepatic expression of caspase-11 was significantly higher in I/R group than in sham group ( < 0.05), and was significantly lower in isoflurane group than in I/R group ( < 0.05). CONCLUSIONS Isoflurane preconditioning has protective effect against hepatic I/R injury, which is related to the inhibition of the noncanonical pyroptosis pathway.
Animals ; Caspases, Initiator ; Ischemic Preconditioning ; Isoflurane ; Liver ; Mice ; Mice, Inbred C57BL ; Pyroptosis ; Reperfusion Injury

Objective Laparoscopic splenectomy has become an effective method to treat the hypersplenism and splenomegaly caused by various diseases.Although laparoscopic technology is very mature at present,laparoscopic splenectomy is still associated with many complications.This study was based on the comparison of different processes and operation techniques of laparoscopic splenectomy,to summarize a more effective operation process and operation technique which can reduce the related complications of laparoscopic splenectomy.Methods The study reviewed the clinical data of 130 patients'who underwent laparoscopic splenectomy in our department from September 2015 to December 2017.According to different operation mode and operation technique these patients were divided into 6 groups.Operation process groups:group Ⅰ for arterial priority group (open the gastrocolic ligament and expose splenic artery,then dissociate ligament around the spleen,and finally divide from the spleen pedicel) had 73 cases;group 2 with the way of building a tunnel beneath spleen pedicel and dissecting spleen in situ had 57 cases.With regard to the way of breaking spleen pedicel:group 3 using stapler to break spleen's pedicel included 101 cases;group 4 using dissecting the secondary spleen pedicel included 29 cases.Whether blocking the splenic vein:group 5 whose splenic vein was not ligated had 95 cases;group 6 whose splenic vein was ligated coverd 35 cases.Analysis and comparison between groups about intraoperative and postoperative complications related to different groups were made.Measurement date were expressed as (-x ± s),t test was used for comparison between groups.Count data was expressed as rate(％),and chi-square test was used for comparison between groups.Results Group 2 has less blood loss [group 1:(356.0 ± 96.2) ml versus group 2:(220.9 ± 57.3) ml,P =0.016],shorter operation time [group 1:(156.0 ± 36) min versus group 2:(120.9 ± 55.4) min;P =0.038] and fewer complications [group 1:9.6％ (7/73) versus group 2:5.3％ (3/57);P =0.035].When compared with group 4,group 3 have more advantages in the intraoperative blood loss [group 3:(256.0 ± 85.1) ml versus group 4:(380.9 ± 107.3) ml;P=0.019],the operation time [group 3:(131.0±30.9) min versus group 4:(171.9 ±61.2) min;P=0.024],and the postoperative complications [group 3:6.95 ％ (7/101) versus group 4:10.3 ％ (3/29);P =0.048].When compared with group 5,group 6 has more advantages in the intraoperative blood loss [group 5:(250.0 ± 56.2) ml versus group 6:(320.9 ± 87.3) ml;P =0.012],the operation time [group 5:(136.0 ± 36.2) min versus group 6:(180.9 ± 57.3) min;P =0.013],and the postoperative complications [group 5:6.3％ (6/95) versus group 6:11.4％ (4/35);P=0.013].Conclusion Both the procedure of tunnel procedure in situ cutting spleen and the application of stapler can reduce the amount of bleeding,shorten the operation time,and reduce the occurrence of postoperative complications.The ligation of the splenic vein is not necessary because it increases the operation time,the risk of bleeding,and the occurrence of postoperative complications.

Objective To discuss the surgical skills and clinical value of laparoscopic splenectomy with a safe approach to the splenic hilum through first mobilizing the lower pole of the spleen.Methods A total of 88 patients with hypersplenism secondary to liver cirrhosis who underwent laparoscopic splenectomy in our department from September 2015 to September 2016 were selected into this study.And these patients were divided into two groups based on whether to take the safe approach to the splenic hilum through first mobilizing the lower pole of the spleen in laparoscopic splenectomy.The control group included 40 cases who underwent the traditonal laparoscopic splenectomy,while the observation group included 48 cases who took the safe approach to the splenic hilum through first mobilizing the lower pole of the spleen in laparoscopic splenectomy.The clinical data were collected retrospectively by medical clinical records review.Results There was no mortality occurred in this study.The blood loss of the observation group was (247.50±135.89)mL,which was obviously lower than (361.75±144.43)mL of the control group,and the difference was statistically significant (P<0.05).The operation time of the two groups were (194.69±47.99)min and (232.75±45.26)min respectively,and the difference was statistically significant (P<0.05).No significant difference was found in terms of rate of conversion to laparotomy,rate of intraoperative blood transfusion,postoperative hospital stay and complications (P>0.05).Conclusion It is very crucial for a safe approach to the splenic hilum through first mobilizing the lower pole of the spleen to establish a tunnel behind the splenic hilum in laparoscopic splenectomy.It is safe and feasible to perform a safe approach to the splenic hilum through first mobilizing the lower pole of the spleen in laparoscopic splenectomy.The technique is generalized in clinics,especially for freshmen.

OBJECTIVETo compare the efficiency and cytotoxicity of different transfection reagents used in transfection of RIP140-siRNA into Kupffer cells to optimize the transfection conditions.

METHODSKupffer cells were transfected with RIP140-siRNA labeled with GFP as the reporter gene using lipofectamine 2000, Roche reagent (X-treme GENE siRNA Transfection Reagent) and puro screening lentivirus (1.0×10(8) TU/mL) as the transfection reagents. The transfection effect was observed under a fluorescent inverted microscope, and laser scanning confocal microscopy was used to analyze RIP140 expression in trasnfected Kupffer cells. Flow cytometry was performed to detect cell apoptosis, and CCK-8 test was used to evaluate the cell proliferation inhibition. RT-RCR and Western blotting were performed to detect the expressions of RIP140 mRNA and protein in the trasnfected cells.

RESULTSPuro screening lentivirus yielded the highest cell transfection efficiency, which exceeded 90%, followed by Roche reagent and then by lipofectamine 2000. Flow cytometry and CCK-8 test showed that the cytotoxicity was the mildest with Roche reagent, moderate with lentivirus, and severe with lipofectamine 2000. The cells trasnfected with lentivirus showed a significantly lower RIP140 expression than cells trasnfected with lipofectamine 2000 and Roche reagent (P<0.05).

CONCLUSIONIn Kupffer cells, lentivirus-mediated transfection, as compared with the other two trasnfection reagents, can achieve good transfection efficiency with a relativelty low cytotoxicity, and allows for better controllability and stability of the trasnfectiion conditions.

Adaptor Proteins, Signal Transducing ; genetics ; Apoptosis ; Cell Proliferation ; Flow Cytometry ; Humans ; Indicators and Reagents ; chemistry ; Kupffer Cells ; drug effects ; Lentivirus ; Lipids ; chemistry ; Nuclear Proteins ; genetics ; RNA, Messenger ; RNA, Small Interfering ; genetics ; Transfection

Objective To systematically evaluate the comparison of postoperative complications of laparoscopic splenectomy (LS) versus open splenectomy (OS) for hypersplenism secondary to portal hypertension of liver cirrhosis .Methods Clinical trials ,which compared the postoperative complications of LS versus OS for portal hypertension of liver cirrhosis from the PubMed ,Cochrane Li‐brary ,EMbase ,CNKI and CBM data bases from 2000 to 2013 and the RevMan 5 .2 software was used for data analysis .Results six trials with 456 patients were included .Meta‐analysis showed that there was no significant difference in incidences of postoperative bleeding(OR=0 .52 ,95% CI 0 .25 -1 .06 ,P=0 .07)or pancreatic fistual(OR=1 .09 ,95% CI 0 .33 -3 .60 ,P=0 .89) between LS and OS groups .Incidence of postoperative infection was decreased in LS group compared with OS group (OR=0 .17 ,95% CI 0 .07-0 .38 ,P<0 .01) .However ,the incidence of postoperative portal venous thrombosis was increased in LS group compared with OS group(OR=2 .14 ,95% CI 1 .23 -3 .73 ,P=0 .00) .Conclusion LS and OS own the same incidence of postoperative bleeding and pancreatic fistual .The incidence of postoperative infection was decreased in LS group compared with OS group ,meanwhile ,LS group was increased compared with OS group on incidence of postoperative portal venous thrombosis .

Objective To compare the efficiency and cytotoxicity of different transfection reagents used in transfection of RIP140-siRNA into Kupffer cells to optimize the transfection conditions. Methods Kupffer cells were transfected with RIP140-siRNA labeled with GFP as the reporter gene using lipofectamine 2000, Roche reagent (X-treme GENE siRNA Transfection Reagent) and puro screening lentivirus (1.0 × 108 TU/mL) as the transfection reagents. The transfection effect was observed under a fluorescent inverted microscope, and laser scanning confocal microscopy was used to analyze RIP140 expression in trasnfected Kupffer cells. Flow cytometry was performed to detect cell apoptosis, and CCK-8 test was used to evaluate the cell proliferation inhibition. RT-RCR and Western blotting were performed to detect the expressions of RIP140 mRNA and protein in the trasnfected cells. Results Puro screening lentivirus yielded the highest cell transfection efficiency, which exceeded 90%, followed by Roche reagent and then by lipofectamine 2000. Flow cytometry and CCK-8 test showed that the cytotoxicity was the mildest with Roche reagent, moderate with lentivirus, and severe with lipofectamine 2000. The cells trasnfected with lentivirus showed a significantly lower RIP140 expression than cells trasnfected with lipofectamine 2000 and Roche reagent (P<0.05). Conclusion In Kupffer cells, lentivirus-mediated transfection, as compared with the other two trasnfection reagents, can achieve good transfection efficiency with a relativelty low cytotoxicity, and allows for better controllability and stability of the trasnfectiion conditions.

Objective To compare the efficiency and cytotoxicity of different transfection reagents used in transfection of RIP140-siRNA into Kupffer cells to optimize the transfection conditions. Methods Kupffer cells were transfected with RIP140-siRNA labeled with GFP as the reporter gene using lipofectamine 2000, Roche reagent (X-treme GENE siRNA Transfection Reagent) and puro screening lentivirus (1.0 × 108 TU/mL) as the transfection reagents. The transfection effect was observed under a fluorescent inverted microscope, and laser scanning confocal microscopy was used to analyze RIP140 expression in trasnfected Kupffer cells. Flow cytometry was performed to detect cell apoptosis, and CCK-8 test was used to evaluate the cell proliferation inhibition. RT-RCR and Western blotting were performed to detect the expressions of RIP140 mRNA and protein in the trasnfected cells. Results Puro screening lentivirus yielded the highest cell transfection efficiency, which exceeded 90%, followed by Roche reagent and then by lipofectamine 2000. Flow cytometry and CCK-8 test showed that the cytotoxicity was the mildest with Roche reagent, moderate with lentivirus, and severe with lipofectamine 2000. The cells trasnfected with lentivirus showed a significantly lower RIP140 expression than cells trasnfected with lipofectamine 2000 and Roche reagent (P<0.05). Conclusion In Kupffer cells, lentivirus-mediated transfection, as compared with the other two trasnfection reagents, can achieve good transfection efficiency with a relativelty low cytotoxicity, and allows for better controllability and stability of the trasnfectiion conditions.

Hepatocellular carcinoma (HCC) is one of the most common cancers in the worldwide.In recent years,the development of imaging techniques has facilitated the detection of HCC at early stages.However,HCC has a high frequency of portal vein invasion,which is reportedly observed in 11％ to 42％ of patients with HCC.Portal vein tumor thrombus(PVTT) is a crucial factor that can worsen the prognosis of HCC because it can be wide dissemination of tumor throughout the liver and cause a marked deterioration of hepatic function.It was considered that is not appropriate with surgical treatment in the past,which is treatment conservative or even give up,most of the patients died within a few months.Even if the patients have a chance to surgical resection,and the most of patients combined with TACE,portal vein stent implantation or chemotherapy postoperative,some patients can be more prolonged survival,however,due to the small tumor thrombus and micro metastases have exist before the surgery and can not be checked out.it's difficult to clean up in the surgery,easily lead to HCC recurrence postoperative.However,with the constant awareness the PVTT formation mechanism and scientific technology progress,more and more new treatment techniques is applied to clinical.This view focuses on the portal vein tumor thrombus formation mechanisms and Classic Transcatheter Arterial Chemoembolization and new treatment methods such as:Helical tomotherapy,CIK cell therapy (cytokine-induced killer cell therapy),Chemoembolization and stent combined with iodine-125 seed.

At present, about one third organ transplantation recipients were hepatocellular carcinoma (HCC) patients even in the most advanced transplant center in the world. HCC especially the early primary HCC has been listed as one of the indications of liver transplantation(LT) in the most organ transplant center in the world. However, due to the tumor recurrence and metastasis following LT, many patients would be dead following transplantation in about 2 years later. Their five-year survival rate was only 50 percent in benign final phase liver disease patients. The current researches for this phenomenon were still shallow both in mechanisms and treatments. How to prevent tumor recurrence has become serious challenges and must be faced in liver transplantation surgery. Obviously, to discover the molecular mechanisms and predisposing factors of following LT recurrence, to explore appropriate standards for LT in HCC patients and to strengthen perioperative management are the only way for increasing LT efficacy.