Objective To develop a new culture system in which hepatocytes can restore in-vivo like function. Methods Sandwich configuration was used for the culture of hepatocytes, and protein synthesis as well as P 450-b ,albumin mRNA expression by in situ hybridization was studied. ResultsStable and gradually increasing protein synthesis and high level of P 450-b , albumin mRNA expression were observed. In contrast, protein secretion of hepatocytes cultured in conventional single collagen culture system was low and decreased until approximately zero seven days after, and mRNA expression of P 450-b ,albumin was low. Conclusions Sandwich configuration in which cell polarity of hepatocytes is reestablished like in vivo situation.

Objective To reestablish cell polarity of hepatocytes cultured in vitro.Methods Sandwich configuration was used to culture hepatocytes. The morphology as well as specific plasma proteins were compared with a single collagen configuration.Results In sandwich configuration hepatocytes were arranged in single cell plate, and the gradual development of bile canaliculi like structures and an anastomotic network were observed over the course. Histochemical immunolocation revealed that membrane proteins were distributed specifically. Hepatocytes in single collagen configuration showed hardly any hepatic plate like structure,and the distribution of specific plasma protein was not formed. Conclusions Sandwich cultured hepatocytes reestablished cell polarity structurally and regained the characteristics of hepatocytes in situ.

For a matured digestive surgeon,pancreaticoduodenectomy (PD) is regarded as one of the most complicated and technically challenging surgical procedure.Based on the accurate interpretation of patient's preoperative imageologic data,we advocate a novel procedure which is called as G-path pylorus-preserving pancreatoduodenectomy (G-path PPPD).We deen G-path PPPD as a standardized procedure for resectable pancreatic head cancer or periampullary carcinoma,which definitely simplify the procedure,save the operative time,achieve R0 resection through en-bloc resection without interruptedly intraoperative exploration and reduce the risk of iatrogenic tumor metastasis.This article introduced the program of G-path PPPD in detail by taking a patient as an example who suffered from pancreatic head cancer accompanied with obstructive jaundice,and discussed the relevant points.

AIM　To investigate the effect of hydroxyl camptothecinum(HCPT) on pharmocokinetics of cyclosprine A(CsA) and the membrane lipid fluidity (LFU)of red cell in rabbits. METHODS　The whole blood concentration of CsA in rabbits was monitored using method of FPIA. The LFU of red cell was measured using method of DPH fluorescence polarization technique. RESULTS　The pharmocokinetic parameters of CsA in group CsA+HCPT such as a,V(c),K12 and CL(s) were significantly lower and T1/2(a) was significantly higher (P<0.05) than in group CsA. The concentration of CsA of group CsA+HCPT was higher after injection 0.25, 0.5 and 1.0 h, but have significantly high at 1.0 h point (P<0.05). After injection 0.5,1.0, 1.5 h the LFU of red cell in group CsA+HCPT was higher than in group CsA (P<0.05). The LFU in group CsA was lower that in control at 0.5,1.0 and 1.5 h point (P<0.05). CONCLUSION　HCPT can enhance the whole blood concentration of CsA in rabbits. CsA administrated with HCPT is feasible.

AIM To investigate the effect of hydroxyl camptothecinum(HCPT) on pharmocokinetics of cyclosprine A(CsA) and the membrane lipid fluidity (LFU)of red cell in rabbits. METHODS The whole blood concentration of CsA in rabbits was monitored using method of FPIA. The LFU of red cell was measured using method of DPH fluorescence polarization technique. RESULTS The pharmocokinetic parameters of CsA in group CsA+HCPT such as a, V (c), K 12 and CL (s) were significantly lower and T 1/2(a) was significantly higher ( P

Objective To explore the effect of ulinastatin on coagulation function of elderly patients after hip replacement.MethodsTotally 120 patients undergoing elective hip replacement in our hospital were chosen.According to the stratified randomization method, all patients were divided into observation group and control group with 60 in each.The control group received oral Xarelto combined with other conventional treatment, while the observation group received ulinastatin on the basis of the treatment regimen of the control group.The coagulation parameters and clinical manifestations were compared between two groups.ResultsTwo days after operation, the TT, APTT, PT, DD and FIB between the two groups showed statistically significant difference (t=-5.300,-2.319,-2.409,-2.325,-3.567;P<0.05).The hospital stay,CRP,TXB2 and total drainage volume in observation group 5d after operation were lower than those in control group, between the two groups showed statistically significant difference (t=-2.529,-2.082,-3.388,-2.887;P<0.05).The patients with elevated blood urea and serum creatinine in observation group were obviously less than that of control group, but the patients with nausea in observation group were obviously higher than those in control group,between the two groups showed statistically significant difference (χ2=5.217,5.926,8.571;P<0.05).The patients with deep vein thrombosis in observation group were obviously higher than those in control group,between the two groups showed statistically significant difference (χ2=8.571, P<0.05).ConclusionUlinastatin can significantly improve the hypercoagulable state of patients after hip replacement and reduce the incidence of deep vein thrombosis, and plays an important role in promoting wound healing and immunity recovery.

Objective To investigate the effect of dexmedetomidine combined with ulinastatin on postoperative delirium in elderly patients undergoing resection of gastrointestinal tumor. Methods A total of 180 elderly patients (97 males,83 females,aged 65-80 years,ASA grade Ⅱ orⅢ)who underwent laparoscopic surgery for gastrointestinal tumor,were randomized into four groups (n =45 each):dexmedetomidine group (group D),ulinastatin group (group U),dexmedetomidine+ulinastatin group (group DU)and control group (group C).Patients in group D were given a loading dosage of dexmedetomidine 0.5 μg/kg intravenously 1 5 min before the induction of general anesthesia,followed by a continuous infusion of 0.3 μg·kg-1 ·h-1 ,and dexmedetomidine was ad-ministered till 40 min before the end of surgery.Patients in the group U were given a loading dosage of ulinastatin 10 000 U/kg intravenously in 20 min.In group DU,dexmedetomidine and ulinastatin were administered in accordance in group D and group U respectively.Patients in group C were given 0.9% saline solution.The volume of blood loss,the time of operation and recovery,the adverse reac-tions after surgery were recorded.The concentration of dopamine (DA),adrenaline (AD),norepi-nephrine (NE)were measured within the preoperative 1 d (T0 ),within the first hour of surgery (T1 ),within the postoperative 1 d (T2 ),2 d (T3 ),3 d (T4 ).The confusion assessment method Chi-nese reversion (CAM-CR)was used to screen POD on T0 ,T2-T4 .Results The levels of DA,AD and NE in the group C and group U at T1-T4 significantly elevated than those at T0 (P <0.05);the levels of DA at T1 and the levels of AD at T1 ,T2 in group D and group DU significantly elevated than those at T0 (P <0.05).The levels of DA,AD at T3 ,T4 and the levels of NE at T1-T4 in group D and group DU were significantly reduced compared with those in the group C and group D (P <0.05 ). Compared with the group C,the incidence of POD was significantly reduced in the group D,group U and group DU (P <0.05).Among the three groups (D,U and DU),the difference were not statisti-cally significant in the incidence of POD.Conclusion Dexmedetomidine or ulinastatin may reduce the rate of POD in elderly patients undergoing laparoscopic surgery for gastrointestinal tumor.Compared with the administration of ulinastatin or dexmedetomidine alone,combined application of dexmedeto-midine and ulinastatin does not reduce the incidence of POD.

Objective To study the clinical application of laparoscopic cholecystectomy (LC) combined with intraoperative endoscopic sphincterotomy (IOEST) with the antegrade guidewire technique in the treatment of cholecystocholedocholithiasis.Methods This is a prospective controlled study comparing LC combined with preoperative endoscopic sphincterotomy (POEST) versus LC combined with IOEST in the treatment of cholecystocholedocholithiasis.Patients who were diagnosed to have cholecystocholedocholithiasis from June 2012 to February 2013 in our hospital were divided into the POEST group and the IOEST group.There were 50 patients in each group.The operation time,successful stone-extraction rate,residual stones rate,complication rate,postoperative stay and hospitalization cost between the groups were compared.Results The sex,age,stone size,number of stones and diameter of the common bile duct showed no significant difference between the two groups (P ＞ 0.05).There were no differences between the two groups in surgical time of EST,surgical time of LC combined with EST,successful stone-extraction rate and hospitalization cost (P ＞ 0.05).There were significant differences between the two groups in the residual stones rate,postoperative hyperamylasemia rate,postoperative acute pancreatitis rate and postoperative stay (P ＜0.05).LC combined with IOEST using the antegrade insertion of guidewire technique significantly reduced the residual stones rate (0 vs 8％),postoperative hyperamylasemia rate (4％ vs 18％),postoperative acute pancreatitis rate (0 vs 8％) and postoperative stay.Conclusions LC combined with IOEST using the antegrade guidewire technique could be performed safely when compared with the traditional sequential technique,LC combined with IOEST using the antegrade guidewire technique significantly reduced the postoperative acute pancreatitis rate and the residual stones rate.LC combined with IOEST using the antegrade guidewire technique should be the recommended technique to treat patients with cholecystocholedocholithiasis.

Objective To evaluate the effects of sevoflurane on invasion and migration of mouse lung cancer cells induced by hypoxia.Methods Mouse Lewis lung cancer cells were inoculated in the culture plate.After being cultured for 24 h,the cells were randomly divided into 3 groups (n=18 each) using a random number table:control group (group C),hypoxia group (group H) and hypoxia+ 2％ sevoflurane group (group HS).Cells were exposed to 95％ air-5％CO2 (2 L/min) for 4 h in group C.Cells were exposed to 94％ N2-5％CO2-1％ O2 for 4 h in group H.In group HS,cells were exposed to 2％ sevoflurane and 94％ N2 (2 L/min) for 4 h.The invasion of cells was determined by Transwell assay,and the invaded cells were counted.The migration of cells was evaluated by wound healing assay,and cell migration rates were calculated.The expression of Beclin 1 and LC3 Ⅱ protein in cells was detected by Western blot.Results Compared with group C,the number of invaded cells and cell migration rates were significantly increased,and the expression of Beclin Ⅰ and LC3 Ⅱ was up-regulated in H and HS groups.Compared with group H,the number of invaded cells and cell migration rates were significantly decreased,and the expression of Beclin 1 and LC3 Ⅱ was down-regulated in group HS.Conclusion Sevoflurane can inhibit the invasion and migration of mouse lung cancer cells induced by hypoxia,and inhibition of autophagy is involved in the mechanism.

Objective To evaluate the effects of sevoflurane on the expression of heparanase ( HPA) and fascin in lung carcinoma cells of mice. Methods Mouse LLC cells were inoculated in the culture plate. After being cultured for 24 h, the cells were equally and randomly divided into 4 groups using a random number table: control group ( group CC) , 1% sevoflurane group ( group Sev1 ) , 2% sevoflurane group ( group Sev2 ) , and 3% sevoflurane group ( group Sev3 ) . Cells in Sev1-3 groups were exposed to 1%, 2% and 3% sevoflurane, respectively, for 4 h, while cells in group CC were not exposed to sevoflurane, and all the cells were then cultured for another 24 h in an incubator. The invasion of cells was determined by Transwell invasion assay, and the invaded cells were counted. The migration of cells was determined by wound healing assay, and cell migration rates were calculated. The expression of HPA and fascin in cells was detected by Western blot. Results Compared with group CC, the number of invaded cells and cell migration rates were gradually decreased, and the expression of HPA and fascin was gradually down?regulated with increasing concentrations of sevoflurane in Sev1-3 groups. Conclusion The mechanism through which sevoflurane inhibits the metastasis of mouse lung carcinoma cells is associated with down?regulated expression of HPA and fascin.