Objective To study the inhibiting effect of genistein and cisplatin on tumor recurrence and metastasis after hepatectomy in mice. Methods A posthepatectomy high-metastatic-and-recurrent athymic mouse model simulating human HCC was established.Genistein,cisplatin,and combination genistein and cisplafin respectively were given intraperitoneally.Mice were sacrificed after 4 weeks,the volume and pulmonary metastasis of the recurrent tumor was observed.Caculate q value using Jin Zhengjun formula was used to evaluate the synergistic effect of combination genistein and cisplatin.Immunohistochemistry and real time fluorescent quantitation PCR were used to detect the expression of MMP2 and MMP-2 mRNA in liver recurrent tumor. Results Compared with single drug group,mice in genistein combined with cisplatin group had smaller liver recurrent focus volume,less pulmonary metastasis.Genistein and cisplatin displayed additional inhibiting effect on tumor recurrence and metastasis after hepatectomy in vivo,and displayed synergistic inhibiting effect on pulmonary metastasis.The MMP-2 expression of the recurrent tumor in single cisplatin group increased compared with control group(t=26.17、P＜0.05),while in single genistein group and genistein and cisplatin combined group it decreased(t=5.58,13.90,P＜0.05).The expression of MMP-2 mRNA in genistein group was 90%the level of the control group,in cisplatin group it was 2.06 times that of the control group,in combination genistein and cisplatin group it was 44%the level of the control group. Conclusions In vivo,genistein reinforces the effect of cisplatin in inhibiting tumor recurrence and metastasis after hepatectomy,possibly by a mechanism in which genistein inhibits the up-regulation of MMP-2 induced by cisplatin.

Objective To investigate the risk factors of pancreatic fistula after pancreaticoduodenectomy (PD).Methods The clinical data of 186 patients who received PD at the Second Affiliated Hospital of Kunming Medical College from May 2000 to May 2010 were retrospectively analyzed.All patients were divided into pancreatic fistula group (39 patients) and non-pancreatic fistula group ( 147 patients).Risk factors of pancreatic fistula after PD were screened out from 20 factors by univariate and multivariate analysis.The univariate analysis was carried out by chi-square test or Fisher exact test,and the muhivariate analysis was done by Logistic regression.Results Thirty-nine patients were complicated with pancreatic fistula,including 26 in grade A,10 in grade B and 3 in grade C.The results of univariate analysis showed that duration of preoperative jaundicc,loss of body weight at 6 months before operation,preoperative total bilirubin level,preoperative albumin level,postoperative albumin level,length of pancreas dissected,pancreatic tube diameter,pancreatic texture and time of abdominal drainage tube pull out were high risk factors of pancreatic fistula ( x2 =34.990,20.480,8.212,10.890,13.561,11.505,13.820,4.539,36.590,P ＜ 0.05).The results of multivariate analysis showed that duration of preoperative jaundice ＞ 8 weeks,loss of body weight at 6 months before operation ≥ 10％,pancreatic tube diameter ＜ 3 mm,soft pancreatic texture and time of abdominal drainage tube pull out ＞ 5 days were the independent risk factors of pancreatic fistula ( OR =2.229,3.383,1.437,1.273,11.939,P ＜ 0.05).Conclusion Duration of preoperative jaundice ＞ 8 weeks,unconscious loss of body weight ≥ 10％ within 6 months before operation,pancreatic tube diameter ＜ 3 mm,soft pancreatic texture,time of abdominal drainage tube pull out ＞ 5 days would increase the risk of pancreatic fistula after PD.

Objective To explore the rationality of pancreaticojejunostomy and double purse embedded pancreaticogastrostomy in pancreaticoduodenectomy through comparing the short-term effects between pancreaticojejunostomy and double purse embedded pancreaticogastrostomy in pancreaticoduodenectomy.Methods A retrospective review was performed for 38 patients who were underwent pancreaticoduodenectomy at the second affiliated hospital of Kunming Medical University from January 2010 to December 2012.Results All 38 pancreaticoduodenectomy were performed successfully.The pancreatic fistula (0％),operationtime [(327.50±32.56) min],postoperative hospital days [(10.31 ±2.44) d] were found to be significantly lower in double purse embedded pancreaticogastrostomy group than those of pancreaticojejunostomy group [3/12,(425.83 ± 33.77) min,(14.33 ± 4.14) d],which has statistical significance(P＜0.05).Although the incidence of bile leakage,postoperative abdominal bleeding,delayed gastric emptying,infection of incision,infection of lung,intraoperative amount of bleeding are also lower than that of pancreaticojejunostomy group,which has no statistical significance.Conclusions Double purse embedded pancreaticogastrostomy is a digestive tract reconstruction which is safer and more reasonable than pancreaticojejunostomy.The former one is operated simply and the operation time is shorter.There are great advantages in the prevention of complications such as pancreatic fistula after pancreaticoduodenectomy.

Different degrees of HCV re-infection exist in patients with hepatitis C after liver transplantation. Its pathogenesis is different according to different phases of the disease. Factors affecting its recurrence include HCV gene type, viral load, HLA matching between donor and recipient, time of recurrence, donor's age and so on, in which the application of immunosuppressants is the most important influencing factor. The virological response can be used to evaluate the effects of treatment. Now, it is widely accepted that the best choice and therapeutic plan is Pegy interferon alfa-2a/2b combined with Ribavirin.

Objective To explore the use of CA199,CEA,CA50 and ALP combined with magnetic resonance imaging (MRI) in diagnosis of intrahepatic bile duct stones with early bile duct carcinoma.Methods The clinical diagnosis of 36 cases of patients with hepatolithasis-associated intrahepatic cholangiocarcinoma (HICC) and 118 cases with intrahepatic duct stone (IHDS) were analyzed retrospectively.Serum CA199,CEA,CA50,ALP and magnetic resonance (MRI,MRCP) were performed and the results were analyzed.Results Abdominal pain discomfort in Hepatolithasis-associated intrahepatic cholangiocarcinoma showed multiple symptoms.Three typical Charcot syndromes were rare.On the gender distribution,intrahepatic bile duct stones was frequently found in women,on the contrary,Hepatolithasis-associated intrahepatic cholangiocarcinoma was frequently more found in men (P ＜0.05) Abdominal pain and fever in patients of Hepatolithasis-associated intrahepatic cholangiocarcinoma was more than that of patients with Intrahepatic bile duct stone (P＜0.05) When alkaline phosphatase (ALP) was more than 169 U/L,significant difference was seen between two groups (P＜ 0.05) According to the diagnosis of HICC,the accuracy of CA199,CEA,CA50 combined with ALP was 88.6％,the accuracy of magnetic resonance examination alone was 90.2％,and the accuracy of multiple serological markers and magnetic resonance was 95.5％.Conclusion MRI combined with serum CA199,CEA,CA50 and ALP can effectively improve the HICC preoperative diagnosis rate.

Objective To evaluate the therapeutic effects of gemcitabine regional artery perfusion combined with systemic chemotherapy for late-stage cancer of pancreas.Methods Thirteen cases of late-stage cancer of pancreas proven by operation and pathology were treated with 5-FU+MMC as a combined system chemotherapy, and use of gemcitabine for regional artery perfusion chemotherapy.Results Among the 13 patients who could be evaluated for therapeutic effect, four cases had partial response (PR), six cases had (SD), three cases were PD, and the effective rate of the clinical benifical-reflected evaluation was 76.9%, pain releive rate 75.0% the median survival time was 6.3 months. None of the patients have had serious toxious side-effects.Conclusions The gemcitabine regional artery perfusion combined with systemic chemotherapy can relieve the cancer pain of patients with late-stage cancer of pancreas, improve their general condition, increase the survival quality of life, prolong the survival time. The drug tolerance of the patients is good.

Objective To summaize the experience of application of direct digital radiography and electrohydraulic shockwave lithotripsy in treatment of residual bile duct stones.Methods A retrospective analysis of clinical records of 60 patients with residual bile duct stones treated in our hospital from 2003 to 2006 was made.Results In 59(95.9%) of 60 cases the stones were removed completely,including successful removal in one time of application of this method in 57 Patients(95.0 %),and 2 applications in 2 Patients ﹙3.3 %).One case with bile duct stone complicated with dile duct cancer was unsuccessful.The biggest stone was 3.5 cm?2.5 cm?2.0 cm.No serious complications occurred and changed to operation.The 59 cases were followed up for 6 mo to l yr,and all had a successful outcome.Conclusions The method of combination of direct digital radiography and electrohydraulic shockwave lithotripsy in treatment of postoperative residual bile duct stones after operation can facilitate removal of residual bile duct stones.The procedure is a safe way of treating residual stones.

Objective HLA-G widely participates in immune tolerance by its combination with immunoglobulin-like tran-scripts IL-2 and IL-4 on the surface of dendritic cells (DCs).The aim of the article was to explore the effects of recombinant adnovirus-mediated HLA-G transfection in macaca mulatta immature dendritic cells on T cell proliferation . Methods Marrow blood was collected from macaca mulattas by the puncture needle after anesthesia .Density gradient centrifugation method was applied in separating mononuclear from the extracted blood on which CD 34+cells were collected and pu-rified by means of immunomagnetic separation .Small doses of cyto-kines were added to get the immature dendritic cells after induced dif-ferentiation of CD34+cells.After the recombinant adnovirus-mediated HLA-G transfection in macaca mulatta immature dendritic cells , observation was done on the viral infection efficiency and western blot was used in detecting the expression of HLA -G in immature den-dritic cells.Taking T cells in macaca mulatta as responders and DCs transfected by recombinant adnovirus -mediated HLA-G as stimu-lators, mixed lymphocyte test was conducted .T cells were divided into 5 groups: mDC group ( mature DCs ) , imDC group ( immature DCs), imDC(L) group(addition of 100 ng/mL lipopolysaccharide after getting imDC at 7th day) , imDC(V) group (imDCs infected by recombinant adnovirus-mediated HLA-G) , imDC( L+V) group ( imDCs infected by recombinant adnovirus-mediated HLA-G along with the addition of 100 ng/mL lipopolysaccharide in culture process ) . Results We obtained the immature dendritic cells and recom-binant adenovirus of HLA-G expressed in these cells .Flow cytometry showed DC purity was up to 92.3 %, imDC purity was up to 72.39%and positive percentage of CD 4+T was greater than 80%.In comparison with imDC group ,the proliferation of stimulated T cells in mDC and imDC(L) groups was obviously intensified (P<0.01).In comparison with imDC(V) group, the proliferation of stim-ulated T cells in imDC, mDC, imDC(L), and imDC(L+V) groups was obviously intensified (P <0.01).In comparison with imDC(L+V) group, the proliferation of stimulated T cells in mDC and imDC(L) groups was obviously intensified(P<0.01). Conclu sion Im-mature DCs infected by recombinant adnovirus can inhibit the proliferation of T cells effectively .

Objective To observe expression of resistin gene in adipose tissue of SD rats fed with high-fat diet and its correlation to insulin resistance, diabetes mellitus and leptin. Methods Using RT-PCR to mRNA from adipose tissue, the products were obtained, purified, constructed and squenced. Once its squence was verified, semi-quantitative RT-PCR and Northern Blot were employed to study its expressions in adipose tissues of rats with different diets. Oral glucose tolerance test and insulin release test were performed in 2 groups of rats. Results (1) The resistin gene was successfully constructed and sequenced, which was in accordance with the data of genbank. (2) Compared with the control, the expression of resistin in adipose tissue of high-fat diet rats was significantly enhanced, and blood glucose, serum insulin and leptin levels were also elevated in high-fat diet rats (P

To investigate the clinical features, genetic diagnosis, and treatment of a patient with Prader-Willi syndrome(PWS). For a case with clinically suspected PWS, methylation specific PCR(MSPCR)amplification was applied to CpG islands of SNRPN(exon α)gene locus in the 15q11-q13. Furthermore, the diagnosis was comfirmed by the method of bisulfite sequencing PCR(BSPCR). Metabolic status before and after the operation of sleeve gastrectomy were compared. Absence of amplification of paternal allele on chromosome 15q11-q13 was detected in the case by MSPCR, different from the normal control. Results of BSPCR further proved a full methylation of CpG islands in the SNRPN gene locus. Four months after sleeve gastrectomy, systemic metabolic status and ventricular function were improved. MSPCR and BSPCR were both consistent with genetic diagnosis of PWS. Weight loss surgery is expected to be a major therapy of this disease.