Objective To compare the hepatic differentiation potential of human umbilical cord mesenchymal stem cells (UC-MSCs) with bone marrow mesenchymal stem cells (BM-MSCs).Methods UC-MSCs and BM-MSCs derived from passage 3 were induced by IMDM supplemented with 20 μg/L HGF and 20 mg/L α-FGF.The medium was changed twice a week.The concentrations of albumin and urea nitrogen from cultural medium were measured to compare the differentiation ability of the two cells.We also examined the expression of hepatic related genes by real-time RT-PCR.Results UC-MSCs manifested significandy stronger proliferation potential than BM-MSCs.Both UC-MSCs and BM-MSCs could be induced into hepatocyte-like cells.The morphology of UC-MSCs tended to be more mature than BM-MSCs and they had more cytoplasmic granules.After 4 weeks,the levels of albumin and urea nitrogen from the cultural medium of the UC-MSCs group were significantly higher than the BM-MSCs group (P ＜ 0.05).Real-time PCR showed the expressions of four liver related genes CK18,G6P,HGF and ALB in the UC-MSCs group were significantly higher than the BM-MSCs group (P ＜ 0.05).Conclusion UC-MSCs had higher hepatic differentiation potential than BM-MSCs.Thus,UC-MSCs are more suitable than BM-MSCs for tissue engineering in the treatment of end-stage liver diseases.

Objective To observe the effects of hepatocyte-like cells differentiated from human umbilical cord mesenchymal stem cells (HuMSCs) on the liver function of the rats with liver cirrhosis.Methods Carbon tetrachloride was used to prepare rat model of liver cirrhosis.Then the rats in the experimental group received portal vein injection of 1 ml differentiated hepatocyte-like cells (1 × 107) ; the mesenchymal stem cell (MSC) group was injected with the same volume and number of MSCs; the model group was injected with the same volume of saline (NS) ; the normal rats were treated as control group.After transplantation,the rat angular vein blood and liver tissue were obtained for testing.Results One week after transplantation,compared with the model group,levels of serum alanine aminotransferase (ALT),aspartate aminotransferase (AST) and total bilirubin (TBil) in the experimental group significantly decreased (P ＜0.05),while the albumin (Alb) level increased significantly (P ＜0.05).Compared with the MSC group,the level of Alb in the experimental group also significantly increased (P ＜ 0.05),but there were no differences between the two groups of ALT,AST and TBil.4 weeks after transplantation,compared with the model group,levels of serum ALT,AST and TBil in the experimental group also significantly decreased (P ＜ 0.05),while Alb level increased significantly (P ＜ 0.05).Compared with the MSC group,the differences of the levels of Alb,ALT,AST and TBil were all statistically significant (P ＜ 0.05).Real-time PCR test results showed that the expressions of four liver-related genes of the MSC group and experimental group significantly increased comparing with the model group (P ＜ 0.05).And the experimental group showed higher expression level comparing with the MSC group (P ＜ 0.05).Conclusion The differentiated hepatocyte-like cells could improve hepatic function of patients with liver cirrhosis to a certain degree and showed greater advantage than MSC.

Objective To evaluate the relationship between the expression of MMP-2,MMP-9 and invasion of gallbladder carcinoma.Method Seventeen specimens of gallbladder carcinoma,9 cases of gallbladder adenoma and 9 cases of chronic cholecystis were assessed by using immunohistochemical method(S-P method).Results There were no significant difference in MMP-2 and MMP-9 expressions between gallbladder carcinoma and adenoma,but they were significantly higher than that of chronic cholecystis.The expressions of MMP-2 and MMP-9 had no correlation with the histological differentiation or Nevin staging.Conclusion MMP-2 and MMP-9 may play an important role in tumor invasion and metastasis in gallbladder carcinoma.

AIM：The mechanism of tumor necrosis factor-alpha (TNF-α) induced pulmonary artery hypertension(PAH) in endotoxic shock (ES) is not clear. Cholecystokinin-octapeptide (CCK-8) had anti-ES and anti-PAH effects. The impact of CCK-8 on changes in vascular reactivity and endothelial ultrastructure induced by TNF-α was studied. The role of nitric oxide (NO) was preliminarily studied. METHODS：Rabbit pulmonary artery rings were divided into four groups: TNF-α, TNF-α+CCK-8, CCK-8 and Vehicle. The rings were incubated for 2 h, 7 h or 14 h. Relaxative responses to ACh(10-8-10-5 mol/L)， SNP (10-9-10-6 mol/L) and contractile responses to PE(10-8-10-5 mol/L) were generated seperately. The NOS activity of rings was detected and the ultrastructure of endothelium was observed in the groups that incubated for 7 h.RESULTS:The relaxative responses to ACh were not affected by TNF-α and CCK-8 after incubation for 2 h. TNF-α(7 h,14 h) significantly reduced ACh-induced endothelium-dependent relaxation response of pulmonary artery. CCK-8 reversed the effect. CCK-8 itself had no effect on responses of normal pulmonary artery. Contraction to PE and relaxation to SNP were unaffected by TNF-α, CCK-8. The NOS activity increased in the TNF-α and the TNF-α+CCK-8 groups. While no significant difference was obseved between the Vehicle and the CCK-8 groups. Endothelial injury in TNF-α group and alleviated changes in TNF-α+CCK-8 group were observed. CONCLUSION:TNF-α significantly inhibits endothelium-dependent relaxation, which be one of the mechanisms of PAH induced by TNF-α during ES. It was found for the first time that CCK-8 reversed TNF-α induced impairment of endothelium-dependent relaxation and alleviated structural injury of endothelium, which might be one of the mechanisms of anti-PAH effect by CCK-8 during ES. The effects of TNF-α and CCK-8 might be related to NO.

Objective To summarize the experience in application of amputation of secondary structures of splenic pedicle and self-made spleen-removing bag in laparoscopic splenectomy the using 3-hole method. Methods The clinical data of 11 patients receiving the procedure from June 2007 to April 2009 in our hospital were retrospectively analyzed. Results Advantages of the procedure were less bleeding, small wound, quick recovery and no occurrence of postoperative complications. Six cases had slight postoperative pain. All 11 patients were cured. Conclusion The technique of amputation of secondary structures of splenic pedicle is safe and feasible in laparoscopic splenectomy using the 3-hole method.

Objective To investigate the clinical application of Glissonean pedicle transection method in hepatocellular carcinoma. Methods We analyzed the clinical data of 232 cases of hepatocellular carcinoma underwent hepatic resection in our department from April 2009 to April 2013. Eighty-nine cases underwent hepatic resection by Glissonean pedicle transection method, and 143 cases underwent Pringle maneuver. Results There was no significant differences in the operation time and blood loss between the two groups (P > 0.05). However, the postoperative hospital stay time and the postoperative complications in group A were better than those in group B(P<0.05). Levels of ALT and AST in group A increased significantly slowly than those in group B. The 1, 3-year disease-free survival rates in the two groups were 87.4%, 71.1% and 85.1%, 60.0%, respectively. The 1, 3-year survival rates were 93.3%, 73.5% and 93.4%, 65.0%, respectively, with no significant differences (P>0.05). Conclusion Glissonean pedicle transection method has many advantages including effectively preventing the intraoperative bleeding, maximally reserving the remaining of liver function, decreasing the rate of tumor spread and metastasis. Meanwhile, it is a safe and effective technique for hepatectomy without dissection of the hepatic artery, portal vein and bile duct in the hepatoduodenal ligament.

AIM: To investigate the effect of cholecystokinin octapeptide(CCK-8) on the L-arginine-nitric oxide(NO) pathway in rabbit thoracic aortae treated with lipopolysaccharide(LPS). METHODS: The isolated thoracic aortic rings(TARs) from rabbits were incubated with LPS, LPS+CCK or vehicle for 14 h. Then the contractility to phenylephrine(PE) by TARs and the response to L-arginine(L-Arg) by pre-contractile TARs were measured. In addition, we added NO synthase(NOS) inhibitors aminoguanidine(AG)and N ?-nitro-L-arginine(L-NNA) into organ baths to observe the changes of vascular contractility to PE. NOS activity in isolated TARs were also detected. RESULTS: Incubation of TARs with LPS for 14 h resulted in an increase of NOS activity and a reduction of contractility to PE. Treatment with CCK-8 significantly inhibited the increased NOS activity in thoracic aortae and improved the hypocontractility of TARs to the same degree as AG. CONCLUSION: CCK-8 may improve the hypocontractility of TARs induced by LPS by inhibiting the activity of NOS.

AIM and METHODS: To elucidate the mechanism of anti-endotoxic shock of cholecystokinin octapeptide(CCK-8), the effects of CCK-8 on changes in rabbit thoracic aortic reactivities induced by lipopolysaccharides(LPS) in vitro were studied, and the ultrastructure of the endothelial cells was observed under scanning electron microscope. RESULTS: Incubation of thoracic aortic rings(TARs) with LPS(100 mg/L) resulted in an time-dependent impairment of the endothelium-dependent relaxations to acetylcholine(incubation for 3, 7, 14 h), a reduction of contractive response to phenylphrine(incubation for 14 h) and ultrastructural injury in endothelial cells(incubation for 7 h), all of which were alleviated by concomitant incubation with CCK-8(1 mg/L). In contrast, neither the vascular contractions nor the relaxations were affected by CCK-8 (1 mg/L) alone. CONCLUSION: CCK-8 improved the vascular reactivities in the presence of LPS, which may be one of the anti-endotoxic shock mechanisms of CCK.

Objective To compare the long-term efficacy between laparoscopic liver resection and open liver resection to treat small hepatocellular carcinoma.Methods The clinical data of 52 patients with small hepatocellular carcinoma treated from August 2011 to November 2012 were reviewed.Twenty patients underwent laparoscopic liver resection (the laparoscopic group),while the remaining 32 patients underwent open liver resection (the laparotomy group).The preoperative,postoperative and overall survival data between the two groups were compared.Results The data between the two groups before surgery were comparable (all P ＞ 0.05).The differences in tumor size and pathologic type between the two groups did not reach statistical significance (t =1.087,x2 =0.738,all P ＞ 0.05).However,the length of hospital stay in the laparoscopic group was significantly shorter than in the laparotomy group (t =3.363,P ＜ 0.05).Post-procedural complications occurred in no patients in the laparoscopic group,but in 8 patients in the laparotomy group (x2 =5.909,P ＜ 0.05).The cumulative survival rates in the two groups were not statistically signifi cant (P ＞ 0.05),but the recurrence-free survival of the laparoscopic group was significantly longer than the laparotomy group (P ＜ 0.05).The postoperative 1-year disease-free survival was not significantly different (P ＞ 0.05),though the 3-and 5-year recurrence-free survival rates were significantly different (all P ＜ 0.05).Conclusion The long-term overall survival rate of laparoscopic treatment for small liver cancer was similar to open operation,but the recurrence free survival rate was greatly improved.