5 male adults were studied in continuous bed rest for 16 days. The total body fat was measured by modified water displacement method (density method). The loss of body protein was measured by calculating nitrogen balance. The results have shown that during the experimental peroid their body protein decreased by 167 to 376 g and the body fat increased by 1.28?0.38kg. The average caloric requirement was 1958 ? 242 kcal/man/day, 32.3?1.6 kcal/kgBW/day or 36.1?1.4 kcal/kg LBW/day. These results are similar to those provided by other methods such as dietary survey or gas metabolism.

Objective To analyze the role of a key intracellular signaling molecule GSK3β in hepatocyte apoptosis induced by endoplasmic reticulum stress (ERS).Methods Using mouse hepatoma cell lines(Hepa 1) as cell apoptosis model triggered by tunicamycin,an endoplasmic reticulum stress inducer.One hour before Hepa 1 apoptosis induced by tunicamycin,SB216763 specifically inhibited the activity of GSK3β.Living cells/apoptotic cells were detected using acetoxymethyl (AM)/propidium iodide (PI) staining; Furthermore,the measurement of lactate dehydrogenase(LDH) of cell culture supernatant to evaluate the apoptosis.We detect p-GSK3β,GSK3β,the ERS-related protein(GRP78,CHOP and caspase-12) and caspase-3,cleaved caspase-3 protein expression using Western blot.Results Endoplasmic reticulum stress induced by tunicamycin promotes GSK3β activity; Inhibition of GSK3β activity alleviates endoplasmic reticulum stress:the expression of GRP78,CHOP and caspase-12 expression are inhibited.At the same time,GSK3β activity inhibition significantly reduced the endoplasmic reticulum stress-induced apoptosis:compared to cell apoptosis model group,the intervention group of SB216763 showed that the level of LDH decreased significantly,and PI staining of apoptotic cells was also significant reduction.Western blot results showed that the inhibition of GSK 3 β activity reduced reactive cleaved caspase-3 protein.Conclusion GSK3β is an important signaling molecule in the apoptosis pathway induced by endoplasmic reticulum stress ;Endoplasmic reticulum stress promotes hepatocyte apoptosis by mediating GSK3β.

OBJECTIVETo study the role of endoplasmic reticulum stress (ERS) in acute liver failure (ALF) using a mouse model of D-Galactosamine/lipopolysaccharide (D-GalN/LPS)-induced ALF.

METHODSThe ALF model was established by administering intraperitoneal (i.p.) injections of D-Ga1N (700 mg/kg) and LPS (10 mug/kg) to six C57BL/6 mice. Three of the modeled mice were also administered 4-phenylbutyrate (4-PBA; 100 mg/kg i.p.) at 6 hours before the onset of ALF and served as the intervention group. Non-modeled mice served as controls. All mice were analyzed by western blotting and qRT-PCR to determine the expression levels of ERS-related proteins in liver tissue. Liver function was assessed by measuring levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum. Extent of injury to the liver tissue was assessed by hematoxylin-eosin staining and histological analysis. qRT-PCR was also used to detect differences in expression of inflammation-related genes, and western blotting was also used to detect differences in expression of the apoptosis related protein Caspase-3.The extent of apoptosis in liver tissue was assessed by TUNEL assay.

RESULTSThe ERS markers GRP78 and GRP94 showed increased expression at both the gene and protein levels which followed progression of ALF. The ERS effector proteins XBP-1, ATF-6 and IRE 1 a involved in the unfolded protein response were activated in the early stages of ALF, and the ERS-induced apoptosis regulators Caspase-12 and CHOP were activated in the late stage of ALF. Inhibition of ERS by 4-PBA intervention protected against injury to liver tissue and function, as evidenced by significantly lower levels of serum ALT and AST and a remarkably decreased extent of histological alterations. Furthermore, the inhibition of ERS suppressed expression of the proinflammatory cytokines TNFa, IL-6 and IL-1 β, and reduced the extent of hepatocyte apoptosis.

CONCLUSIONERS is activated in the mouse model of D-GalN/LPS-induced ALF. Inhibition of ERS may be protective against liver injury and the mechanism of action may involve reductions in inflammatory and apoptotic factors and/or signaling. Therefore, inhibiting ERS may represent a novel therapeutic approach for treating ALF.

Animals ; Apoptosis ; Disease Models, Animal ; Endoplasmic Reticulum Stress ; Galactosamine ; adverse effects ; Lipopolysaccharides ; adverse effects ; Liver Failure, Acute ; chemically induced ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred C57BL

Objective To investigate the clinical effects of laparoscopic pancreaticoduodenectomy (LPD) for distal cholangiocarcinoma.Methods The retrospective cross-sectional study was conducted.The clinicopathological data of 37 patients who underwent LPD for distal cholangiocarcinoma in the Hunan Provincial People's Hospital between January 2013 and November 2016 were collected.LPD for distal cholangiocarcinoma was performed using the "en-block" procedure.According to the principle of "one axis,two planes and four zones",anatomy used posterior approach,anterior approach and medial approach,lymph node dissection was performed from carotid sheath outside,and en bloc specimens were resected.Observation indicators:(1) surgical and postoperative recovery situations;(2) postoperative pathological examination;(3) follow-up and survival situations.Follow-up using outpatient examination was performed to detect the patients' recurrence-free survival up to November 2017.Measurement data with normal distribution were represented as (x)±s.The survival curve and rate were drawn and calculated by the Kaplan-Meier method.Results (1) Surgical and postoperative recovery situations:all the 37 patients underwent successful LPD,without conversion to open surgery and perioperative death.The operation time,volume of blood loss and case with intraoperative blood transfusion were (326 ± 55) minutes,(176± 39)mL and 1,respectively.The time to initial exsufflation and time for diet intake were respectively (4.5± 1.6)days and (5.3±2.7)days.Of 37 patients,14 had postoperative complications,including 8 with pancreatic fistula (6 with biochemical fistula and 2 with grade B pancreatic fistula),1 with biliary fistula,3 with postoperative bleeding (2 with intra-abdominal bleeding and 1 with gastrointestinal anastomosis bleeding),2 with delayed gastric emptying (grage A),2 with intra-abdominal infection and 2 with pulmonary infection;the same patients can merge multiple complications.Three patients were in Clavien-Dindo classification ≥ Ⅲ.One patient received reoperation and other patients were improved by symptomatic treatment.Duration of hospital stay of 37 patients was 13.5 days (range,8.0-33.0 days).Eight patients underwent adjuvant chemotherapy of 4-6 cycles by taking orally tegafur or gemcitabine with cisplatin.(2) Postoperative pathological examination:tumor diameter and pancreatic duct diameter of 37 patients were (2.1±1.1)cm and (2.5±1.2) mm,respectively.Of 37 patients,9,13 and 15 were respectively detected in high-differentiated,moderate-differentiated and lowdifferentiated adenocarcinoma.Surgical margins:35 patients received R0 resection and 2 received R1 resection.Number of lymph node dissected,cases with lymph node metastasis and number of positive lymph nodes were respectively 18.5±4.9,16 and 1.7± 1.4.Analysis of lymph node metastasis location showed that the positive rates in 8a,12,13,14 and 17 groups lymph nodes were respectively 5.4％ (2/37),18.9％ (7/37),21.6％ (8/37),8.1％ (3/37) and 10.8％ (4/37);Perineural invasion (PNI),lymphovascular invasion (LVI),pancreatic invasion and duodenal invasion were identified in 14,9,16 and 6 patients,respectively.TNM stage:stage 0,Ⅰ A,Ⅰ B,ⅡA and Ⅱ B were respectively detected in 1,3,5,12 and 16 patients.(3) Follow-up situation:of 37 patients,36 were followed up for 6-45 months,with a median time of 26 months.The median recurrence-free survival time,1-and 3-year recurrence-free survival rates were respectively 28 months,80.6％ and 42.2％.Conclusion LPD is safe and effective for distal cholangiocarcinoma,and "en-block" resection not only helps to optimize the process of LPD for distal cholangiocarcinoma,but also has a significant effect on R0 resection and lymph node dissection.

Objective To investigate the causes and countermeasures of reoperation following laparoscopic pancreatoduodenectomy.Methods The causes,approaches and outcome of reoperation were retrospectively analyzed in 10(4.0％,10/250) patients undergoing reoperations following pancreaticoduodenectomy with various complications in Hunan Provincial People's Hospital from April 2014 to April 2018.Results The causes of the 10 patients including intra-abdominal bleeding of seven cases (2 cases combined with pancreatic fistula,1 case with pancreatic and biliary fistula),1 patient with gastrointestinal anastomosis output perforation,1 patient with intra-abdominal abscess,and 1 case with postoperative pancreatitis.The time of reoperation was one day to 82 day after the first operation.The main methods of reoperation including suture and hemostasis,rebuilding the digestive tract,gastrostomy and enterostomy combined with abdominal cavity drainage.The mortality of reoperation following laparoscopic pancreatoduodenectomy was 20.0％ (2/10).Conclusions Intra-abdominal hemorrhage,pancreatic fistula and intra-abdominal abscess are the major causes of reoperation after laparoscopic pancreatoduodenectomy.Timely and decisive reoperation is an effective means to reduce postoperative morbidity and mortality after LPD.