Objective:To construct a recombinant adenovirus containing cytosine deaminase( CD ) gene and thymidine kinase( TK ) fusion gene for the gene therapy research of malignant tumors. Methods: A recombinant cosmid containing CD and TK fusion gene was constructed, and then mixed with DNA TPC and co transfected to the 293 cells by calcium phosphate coprecipitation. Results: The results of restriction and PCR showed the insertion was right and the recombinant adenovirus generated contained the CD and TK fusion gene without replication competent adenovirus. Conclusion: The recombinant adenovirus generated is E1 and E3 deleted and contains double suicide gene needed, which can be further studied for gene therapy.

Objective Intrahepatic biliary papillomatosis(IBP)is a rare disease that is characterized by multiple numerous papillary adenomas in the intrahepatic biliary duct(IBD).The clinical features and outcome,however,are not well known.The clinicopathologic features,treatments and follow-up results were retrospectively analyzed in order to improve the efficiency of diagnosis and treatment for the disease.Methods Between August 2006 and October 2008,9 patients were diagnosed with IBP by histological findings at a tertiary referral center,Ningbo Medical Center(University of Ningbo,College of Medicine,Ningbo,China).The authors retrospectively reviewed the medical records to obtain clinical,radiological and pathologic data.The therapeutic results and follow-up data were also reviewed.Results The ratio of male to female was 2:7 and the middle age was 59 years.Repeated episodes of fever and right upper quadrant abdominal pain with or without jaundice were the common clinical manifestations.There were intrahepatic choledocholithiasis and/or history of previous biliary operation in 8 cases.The level of CA19-9 and CEA were almost normal.Imaging workup demonstrated cyst-like dilatation of intrahepatic biliary tree with or without liver atrophy,which were more obvious in the mucin-hypersecreting IBP.All of the 9 cases underwent curative resection with an adequate resection margin.Macroscopic findings demonstrated that the tumors of 9 cases were all located in the left lobe with mucin-hypersecreting type in 7 cases.The diameter of numerous papillary granular was usually less than 10 mm(from 2 to 5 mm.frequently)and friable,that filled the dilated IHD dispersive.Fine fibrovascular cores lined by dysplastic epithelial cells were frequently found under microscope.Few foci of stromalinvasion were noticed in the two cases with malignant transformation.All of the 9 cases survived and there was no recurrence.Conclusion IBP occurs more often in middle and old women who have history of biliary disease. Repeated episodes of cholangitis are the common clinical manifestations. Extra- and intrahepatic biliary tract dilatation is the common imaging finding.MRCP/ERCP is more valuable than others in diagnosis. Clinical and histological finding shows that IBP is a premalignant disease with high malignant potential. Curative resection should be done as earlyas possible for the long-term survival rate. The use of cholangioscopy in operation and rapid biopsy of resection margin will benefit the curative resection.

OBJECTIVETo compare the difference in depression relief in the treatment of depressive disorder at the acute stage between the combined therapy of acupuncture and 5-HT (5-hydroxytryptamine) selective serotonini reuptake inhibitors (SSRIs) and the single application of SSRIs and explore the impact on the imbalance of 5-HT and TH1/TH2.

METHODSNinety cases of depressive disorder at the acute stage were randomized into a combined therapy group and a medication group, 45 cases in each one. In the medication group, SSRIs were prescribed forl oral administration, once or twice a day, continuously for 4 weeks. In the combined therapy group, on the basis of treatment as the medication group, acupuncture was combined. The main acupoints were Baihui (GV 20), Yintang (GV 29), Shenting (GV 24), Fengchi (GB 20), Dazhui (GV 14) and Sishencong (EX-HN 1), once every two. days, continuously for 4 weeks. Before treatment, and after the 1st, 2nd and 4th weeks of treatment, the Hamilton depression scale (HAMD) was used to evaluate the depression severity. Separately, before and after the 4 weeks of treatment, the levels of serum 5-HT, interleukin-1 beta (IL-1β), interleukin-6 (IL-6), interleukin-4 (IL-4) and interleukin-10 (IL-10) were determined and compared with those in 45 cases of the healthy group.

RESULTSHAMD score was reduced in the 1st, 2nd and 4th weeks of treatment as compared with that before treatment in the combined therapy group (all P<0 01). HAMD score was reduced in the 2nd and 4th weeks of treatment as compared with that before treatment in the medication group (all P<0. 01). HAMD scores in the combined therapy group were lower than those in the medication group in the 1st, 2nd and 4th weeks of treatment (all P< 0. 01). Before treatment, in the combined therapy group and the medication group, the levels of serum 5-HT, IL-4 and IL-10 were all lower than those in the healthy group (all P<0. 01); the levels of IL-1β and IL-6 were higher than those in the healthy group (all P<0. 01). In the combined therapy group and the medication group, the levels of 5-HT, IL-4 and IL-10 in 4 weeks of treatment were all increased as compared with those before treatment (all P<0. 01), and the levels of IL-1β and IL-6 were lower than those before treatment (all P<0. 01). In the combined therapy group, the levels of IL-1β and IL-6 in 4 weeks of treatment were lower than those in the medication group, and the levels of 5-HT, IL-4 and IL-10 were higher than those in the medication group (P<0. 01, P< 0. 05).

CONCLUSIONThe combined therapy of acupuncture and SSRIs achieves much quicker and more effective re-' sult for relieving depression in the patients of depressive disorder as compared with simple oral administration of' SSRIs, and much more contributes to adjust the imbalance of serum 5-HT and TH1/TH2.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Antidepressive Agents ; administration & dosage ; Combined Modality Therapy ; Depressive Disorder ; blood ; drug therapy ; therapy ; Female ; Humans ; Interleukin-10 ; blood ; Interleukin-1beta ; blood ; Interleukin-4 ; blood ; Interleukin-6 ; blood ; Male ; Middle Aged ; Serotonin ; blood ; Th1 Cells ; metabolism ; Th2 Cells ; metabolism ; Young Adult

ObjectiveTo study the efficacy of transcatheter arterial chemoembolization (TACE) after liver resection for hepatocellular carcinoma (HCC) with tumor thrombus in the main trunk and/or first branch of portal vein,and to clarify prognostic factors affecting survival.Methods From 2005 to 2009,there were 358 consecutive patients with HCC who underwent surgical resection in our Department.In 55 patients (15 ％),portal vein tumor thrombus (PVTT) was found intraoperatively or postoperatively during histopathological examinations to involve the first portal branch,main portal trunk,or contralateral portal branch.In this retrospective study,these 55 patients were divided into two groups:Group A,29 patients received postoperative TACE,and Group B,26 patients who did not receive TACE.The clinical data and survivals were compared between the two groups.Prognostic factors were indentified using univariate analysis,followed by multivariate regression analysis using the Cox proportional hazards model.ResultsThere were no significant differences in the demographic clinical data between Group A and Group B.The overall 1-,2- and 3-year survivals for the 55 patients were 63.3 ％,51.4 ％ and 43.5 ％,respectively.The accumulative 1-,2- and 3-year survivals for group A were 71.4 ％,60.1 ％ and 50.1 ％,respectively.The corresponding figures for group B were 56.7％,21.7％ and 10.4％,respectively.Multiple tumors,intrahepatic metastases,hepatic vein thrombus,and invasive type of tumor thrombus were found to be risk factors for short-term survival on univariate analysis,while the latter 3 factors were further found to be significant prognostic factors in the Cox proportional hazards model.Postoperative TACE was shown to be a significant factor in both univariate and multivariate analyses.ConclusionLiver resection was beneficial for some patients with portal vein tumor thrombus.Postoperative TACE further improved the prognosis and prolonged survivals in these patients.

OBJECTIVETo investigate the value and safety of the surgery with vascular resection and reconstruction during pancreatectomy for pancreatic cancer.

METHODSThe clinical data of 206 patients with pancreatic cancer who underwent radical resection were retrospectively analyzed from January 2009 to March 2014 in Lihuili Hospital, Medical center of Ningbo.All cases were divided into non-vascular resection group(132 cases), the combined vein resection group(66 cases) and the combined arterial resection group(8 cases). The peri-operation data, the incidence of postoperative complications and the survival were compared in pairs among three groups.All patients were followed up till September 2014.

RESULTSThere were no statistical differences for the preoperative data among three groups.The operation time and the blood loss (M(QR)) were (347±96)minutes and (500(400)) ml in non-vascular resection group, (425±91)minutes and (800(500))ml in combined vein resection group, (508±120)minutes and (1 750(2 075))ml in combined arterial resection group, with significant differences among three groups(all P<0.01). The incidence of postoperative complication was 16.7%(22/132) in non-vascular resection group, 28.8%(19/66) in combined vein resection group, and 6 cases in combined arterial resection group, respectively.There were significant differences between non-vascular resection group and combined vein resection group(P<0.05), non-vascular resection group and combined arterial resection group(P<0.05), as well as between combined vein resection group and combined arterial resection group(P<0.05). The median survival time was 15 months for non-vascular resection group, 15 months for combined vein resection group, and 12 months for combined arterial resection group.No significant difference was found among three groups(all P>0.05). The postoperative mortality was nil for all of groups.

CONCLUSIONSCompared with non-vascular resection, combined vein resection can be performed safely with a similar prognosis. The surgery of combined arterial resection could only be justified when R0 resection for pancreatic cancer could be achieved for highly selected patients.

Arteries ; surgery ; Humans ; Pancreatectomy ; methods ; Pancreatic Neoplasms ; surgery ; Postoperative Complications ; Prognosis ; Retrospective Studies ; Veins ; surgery

Objective: To investigate the role of piperine on the transformation of endothelial cells into fibroblasts. Methods: Cultured human umbilical vein endothelial cells (HUVECs, 4-6 passage) were used for the main experiments. The transformation models of endothelial cells into fibroblasts were induced by transforming growth factor β (TGF-β) stimulation. HUVECs were divided into 6 groups: control group, TGF-β group and 4 groups treated with various concentrations of piperine (1, 5, 10, 20 μmol/L). CKK-8 was used to detect cell proliferation. The CD31/α-smooth muscle actin (α-SMA) expression level was detected by fluorescent staining. The vascular endothelial cadherin (VE-cadherin)/vimentin expression was detected by immunofluorescence staining. RT-PCR was used detect the mRNA expressions of transformation markers. Western blot was used to detect the protein expression of snail and twist. Results: TGF-β increased HUVECs proliferation (P<0.05), which could be significantly inhibited by 10 and 20 μmol/L of piperine, but not by 1 and 5 μmol/L of piperine. Immunofluorescence results demonstrated that TGF-β increased HUVECs transformation to fibroblasts as shown by downregulated expression of endothelial markers CD31, VE-cadherin, and upregulated expression of α-SMA and vimentin, again, these effects could be attenuated by 10 and 20 μmol/L piperine. The expression levels of collagen type Ⅰ and type Ⅲ were significantly higher in TGF-β group than in control group (P<0.05), significantly lower in TGF-β+10 μmol/L piperine group and TGF-β+20 μmol/L piperine group than in TGF-β group (P<0.05).In addition, RT-PCR results showed that TGF-β increased mRNA expression of transformation markers (snail1, snail2, twist1, twist2), while 10 and 20 μmol/L of piperine could significantly downregulated the mRNA expressions of these markers. The protein expression levels of snail and twist were significantly higher in TGF-β group than in control group (both P<0.05), which was significantly lower in TGF-β+20 μmol/L piperine group than in TGF-β group (both P<0.05). Conclusions Piperine can inhibit the transformation of endothelial cells into fibroblasts. This effect might be viewed as one of the potential mechanisms of reduced myocardial fibrosis post piperine treatment.

Objective:To analyze the prognostic factors affecting salvage liver transplantation (SLT).Methods:The clinical data of 97 patients undergoing liver transplantation in the Ningbo Medical Centre Lihuili Hospital from January 2012 to May 2022 were retrospectively analyzed, including 84 males and 13 females, aged (53.6±7.4) years. Among them, 33 patients underwent primary liver transplantation (PLT) and 64 underwent SLT. SLT patients were subdivided into the groups within the Milan criteria (SLT-A, n=35) and beyond the Milan criteria (SLT-B, n=29), according to whether the Milan criteria were met at first hepatectomy. Clinicopathological and prognostic data were subsequently analyzed. Results:The tumor number ( χ2=16.03, P<0.001), microvascular invasion (MVI) ( χ2=10.97, P=0.004), recurrence rate ( χ2=9.31, P=0.010), recurrence-free survival (RFS, F=14.05, P=0.001) and overall survival (OS, F=17.27, P<0.001) were significantly different among the three groups. RFS ( P=0.047) and OS ( P=0.012) in PLT group were better than those in SLT-B group. RFS ( P=0.007) and OS ( P=0.024) in SLT-A group were also better than those in SLT-B group. The multivariate analysis indicated that beyond the Milan criteria at first hepatectomy was an independent risk factor for RFS ( HR=4.378, 95% CI: 1.393-13.756, P=0.011) and OS ( HR=5.391, 95% CI: 1.428-20.352, P=0.013) in patients undergoing SLT, and MVI positive ( HR=4.042, 95% CI: 1.137-14.368, P=0.031) was an independent risk factor for RFS in patients undergoing SLT. Conclusion:Patients beyond the Milan criteria at first hepatectomy and MVI positive showed a poorer prognosis after SLT. Whether the Milan criteria should be the gold standard for SLT as well as for PLT needs further study.