Objective To study the therapy for xenograft human bile duct cancer in mice mediated by adenovirus containing Apoptin gene.Methods Subcutaneous human bile duct cancer was established in nude mice.Variations of tumor volume and histomorphology,side effects were observed after intratumoral injection of adenovirus containing Apoptin gene.Finally the mice were sacrificed for calculating the ratio of antitumor.Results Twelve days after treatment,the mean volume of the xenograft human bile duct cancer in the group of intratumoral injection of adenovirus containing Apoptin gene was(92.31?28.31)mm,which was reduced significantly compared with that of adenovirus infection without apoptin gene(288.86?113.13)mm and control group(344.86?113.87)mm.The ratio of antitumor was 72.10%,which was significantly higher than that in control group(11.9%).During the whole experimental course,no side effect was observed.The histological results demonstrated that the reduction of tumor growth was the result of apoptosis in bile duct cells,which was reduced by transfection of Apoptin gene.Conclusion The adenovirus vectors containing Apoptin gene may constitute a safe tool for the treatment of cholangiocarcinoma.

Objective To observe the effect of fitting rigid gas permeable lenses(RGP) correcting ametropia of keratoconus.Methods 25 patients(43 eyes) with keratoconus fitted RGP,and patinets' visual acuity(VA),cornea condition,degree of comfort and complications were observed.Results All of 43 eyes were successfully to fit RGP at the first time with mean diopter-8.99±-5.55 DS,corrected vision of RGP was significantly better than(or equal to) spherical-cylindrical lens,and get better corrected vision and fewer and more mild complications.Conclusion RGP can get a better correcting vision to keratoconus than spherical-cylindrical lens,and is the good nonoperative treatment to correct ametropia of keratoconus.

Objective To study the influence of cholangiocarcinoma cells on endothelial cells in a co-culture system. Methods A co-culture system of eholangioeareinoma cell line QBC939> and endothelial cells was established in vitro (co-culture group). Endothelial cells were cultured individually during the same time (control group). The mixed supematant of cholangiecareinoma cells and endothelial cells was in the mixed group. Light microscopy and transmission electron micrescopy were used to observe the morphology of the endothelial cells. Changes in expression of ppI25FAK, MMP-2, MMP-9 and uPA of the endothelial cells were detected by mmunofluorescence, and the activities of MMP-2 and MMP-9 were detected by gelatin zymography. All the data were analyzed by paired t test. Results The intercellular space between endothelial cells in co-cuhure group was wider than in the control group. The expression of pp125FAK, MMP-2, MMP-9 and uPA was 394 ±51,455±82, 377±48,422±55 in control group, and was 1096±128,931±72,815±76,801±56 in the eo-euhure group. The difference between the 2 groups had statistical significance (t = 6.53,4.32, 3.61,3.45, P < 0. 05). The values of gray-scale scanning of MMP-2 and MMP-9 in the mixed group were 240.2±15.2 and 2.4±0.8, respectively. The values of gray-scale scanning of MMP-2 and MMP-9 in the co-culture group were significantly increased, they were 687.4 ± 43.6 and 150.9 ± 13.2, respectively (t = 4.89, 5.43, P < 0.05). Conclusions The intercellular space between endothelial ceils and the expression of the proteolytie enzymes are increased after co-culturing endothelial cells with eholangiocarcinoma cells. Proteolytie enzymes may be involved in the process of degradation of subendothelial matrix, and promotes the metastasis of cholangiocarcinoma.

Objective To construct a lentiviral vector of RNA interference (RNAi) of murine triggering receptor expressed on myeloid cells-1 (TREM-1) gene and to explore the effect of TREM-1 on the inflammatory response caused by Bacteroides fragilis. Methods Four target sequences were selected according to murine TREM-1 mRNA sequence, and then 4 pairs of double-strand DNA oligo according to these target sequences and one pair of negative control double-strand DNA oligo were designed and synthesized. These fragments were subcloned into pGCSIL-GFP/Lenti plasmid. After being identified by PCR and sequencing, these plasmids were cotransfected into 293T cells to package lentiviral particles. The lentiviral vector particles were transfected into Raw 264. 7 cells and TREM-1 expression in the transfected cells was assayed by real-time PCR and ELISA. Different concentrations of Bacteroides fragilis lipopolysaccaride (LPS) were administered in the Raw264. 7 cells, and the cells were stimulated with LPS for 12 h. TREM-1 expression was determined by real-time PCR and ELISA at the time points. Results PCR and sequencing confirmed that lentiviral vectors had the correct structure and could express high titer of virus. After being transfected into Raw264. 7 cells, TREM-1 expression was knocked down significantly by all of these lentiviral vectors at both protein and mRNA levels, and the pGCSIL-GFP/Lenti-1 had the most efficient interference. TREM-1 was upregulated in the presence of Bacteroides fragilis LPS, and this increase was partly abrogated in the TREM-1 siRNA-treated cell models of endotoxemia, depending on the sequence. Conclusion The lentivirus RNAi vector of TREM-1 was constructed successfully. The lentivirus RNAi vector of TREM-1 can inhibit the expression of TREM-1 in the murine endotoxemia model caused by Bacteroides fragilis LPS.

Surgical resection is considered to be the most effective therapy for hilar cholangiocarcinoma.Inadequate excision range is the main reason for recurrence after surgery.Extended radical resection provides better long-term survival,however,it may also increase the risk of liver failure because of the extensive hepatic resection.In present study,we showed a new operation which could excise enough length of bile ducts and avoid large volume hepatic tissue resection.The excision extension includes:segment Ⅰ,Ⅳb and partial Ⅳ,left,right and furcation of hepatic duct,extrahepatic ducts,skeletonization of hilar vessels,and dissection of at least second station lymph nodes.As the tissue resected resembles a dumbbell,this surgical technique is named dumbbell type radical resection.The operative indications include:(1) hilar cholangiocarcinoma,Bithmuth Ⅱ and Bithmuth Ⅲ with tumor limited in left or right hepatic ducts ; (2) Without portal invasion; (3) Without third station lymph node metastasis; (4) Without liver or distant organ metastasis.Twenty-three patients had undergone this operation sucessfully.Most patients have high total bilirubin levels (more than 300 μmol/L) and have not received percutaneous transhepatic cholangial drainage or biliary drainage.The average operation time was 355 minutes,and average volume of blood loss during operation was 350 ml.The total survival rate was 65.2％.One-year tumour free survival rate was 95.7％ (22/23),and three-year tumor free survival rate was 7/15.The results indicated that dumbbell type radical resection was feasible for hilar cholangiocarcinoam of Bismuth Ⅱ and Bismuth Ⅲ with tumor limited in left or right hepatic ducts.

Objective To explore the feasibility and clinic outcome of the modified latissimus dorsi flap when it is used to repair huge soft tissue defects in lower limbs.Methods The latissimus dorsi muscle is rich blood supply and available for harvest huge areas.According to these characteristics the modified latissimus dorsi flap was designed:a limited latissimus dorsi skin flap with a large area of latissimus dorsi muscle flap,skin graft area was transferred from donor-site to recipient-site.From April 2009 to August 2011,ten patients with large soft tissue defects in lower limbs were treated with modified latissimus dorsi flap.The size of skin and soft tissue defects range from 19 cm × 10 cm to 32 cm × 16 cm.Ten modified latissimus dorsi flaps had been used for coverage of these wounds,which were amplified 1-2 cm compared with the corresponding wounds.The size of skin flap was 20 cm × 6 cm-33 cm × 10 cm which was allowable to get direct closure of donor-site wounds.Results All the flaps and skin graft survived completely with no complication.Donor-site and recipient-site were primary closed and healing in all patients.All of the patients had got follow-up from 6 months to 36 months.All flaps survived with excellent color and quality,and no extremely fat contour.The results in donor site were satisfying and only left liner scar.It has no effect in the shoulder function.Conclusion The modified latissimus doris flap can provide with a huge amount of skin and soft tissue and be designed in various ways.It is a safe and reliable way to reconstruct huge skin and soft tissue defect in lower limbs.

Objective To investigate the efficacy of different radical surgical procedures for the treatment of hilar cholangiocarcinoma.Methods The clinical data of 207 patients with hilar cholangiocarcinoma who were treated at the Southwest Hospital from June 2007 to June 2012 were retrospectively analyzed.Local resection or hemihepatectomy combined with caudate lobectomy was applied to patients with Bismuth type Ⅰ hilar cholangiocarcinoma; dumbbell type radical resection was applied to patients with Bismuth type Ⅱ hilar cholangiocarcinoma or some patients with type Ⅲ a,Ⅲ b and Ⅳ hilar cholangiocarcinoma; hemihepatectomy or extended hemihepatectomy combined with caudate lobectomy was applied to patients with Bismuth type Ⅲ a,Ⅲ b and Ⅳ hilar cholangiocarcinoma.The patients were followed up every 3 months postoperatively till December 2012.All data were analyzed using the chi-square test or Fisher exact probability test,the survival curve was drawn by Kaplan-Meier method,and the survival was analyzed using the Log-rank test.Results Of the 207 patients,124 received radical resection,including 14 received local resection,23 received dumbbell type resection,87 received lobectomy + caudate lobectomy,49 received palliative resection; 34 received biliary drainage.Four patients died perioperatively.The incidences of complications of dumbbell type radical resection,left hemihepatectomy + caudate lobectomy,right hemihepatectomy + caudate lobectomy were 21.7％ (5/23),46.6％ (27/58) and 48.3％ (14/29),respectively.The incidence of complications after dumbbell type radical resection was significantly lower than left hemihepatectomy + caudate lobectomy and right hemihepatectomy + caudate lobectomy (x2 =4.42,3.90,P ＜ 0.05).One hundred and seventy patients were followed up.The median survival time of the 112 patients who received radical radical resection was 26.5 months,and the 1-,3-,5-year survival rates were 75.9％ (85/112),42.9％ (24/56) and 28.9％ (11/38),respectively.The median survival time of the 38 patients who received palliative resection was 8.5 months,and the 1-,3-year survival rates were 31.6％ (12/38) and 0.The survival time of 20 patients who received biliary drainage was 4.0 months,and the l-year survival rate was 0.The survival rate of patients who received radical resection was significantly higher than those who received palliative resection (x2=65.32,P ＜ 0.05).There was a significant difference in the survival rate between patients who received surgical treatment and those who received biliary drainage (x2=99.97,P ＜ 0.05).Of the 112 patients who received radical resection,the median survival time of 10 patients who received local resection of tumor was 47.0 months,the 1-year survival rate was 10/10,and 4 patients survived at the end of the follow-up; the median survival time of 23 patients who received dumbbell type radical resection was 32.0 months,and the 1-,3-year survival rates were 95.7％ (22/23) and 7/15,and the survival time of 6 patients was longer than 5 years; the median survival time of 54 patients who received left hemihepatectomy or extended left hemihepatectomy + caudate lobectomy was 27.6 months,and the 1-,3-year survival rates were 42.1％ (24/57) and 38.7％ (12/32),and the survival time of 9 patients was longer than 5 years,3 patients survived at the end of the follow-up ; the median survival time of 25 patients who received right hemihepatectomy or extended right hemihepatectomy + caudate lobectomy was 28.3 months,and the 1-,3-year survival rates were 45.8％ (11/24) and 6/15,and the survival time of 6 patients was longer than 5 years,2 patients survived at the end of follow-up.The median survival time of 35 patients (patients with Bismuth type Ⅰ,Ⅱ hilar cholangiocarcinoma and Bismuth Ⅲ a and Ⅲ b hilar cholangiocarcinoma which did not invade the secondary bile duct) who received hemihepatectomy + caudate lobectomy was 32.0 months,and the 1-,3-,5-year survival rates were 91.4％ (32/35),45.8％ (11/24) and 5/16,which were not different from the survival rate of patients who received dumbbell type radical resection (x2 =0.17,P ＞ 0.05).The 5-year survival rate of patients with lymph node metastasis was 4/19,which was significantly lower than 30.4％ (7/23) of patients without lymph node metastasis (x2 =23.40,P ＜ 0.05).Conclusion Joint lobectomy and standardized lymph node dissection could help to improve the efficacy of surgical treatment for patients with hilar cholangiocarcinoma.

Objective To investigate the clinical significance of multiple tumor markers in the early diagnosis of cholangiocarcinoma.Methods 165 patients with histopathologically confirmed cholangiocarcinoma and 25 patients with gallstone canceration were enrolled in this study.36 patients with gallstones and 46 patients with bile duct stenosis were enrolled as controls.The determinations of multiple tumor markers,including carcinoembryonic antigen (CEA),CA125,CA153,CA242,CA19-9,alpha-fIetoprotein (AFP) and Ferritin,of all subjects and the imageological determinations of all the patients with cholangiocarcinoma and with gallstone canceration were analysed retrospectively.Comparisons between the groups were performed using the likelihood ratio test of the Chi-square test.Results CA19-9 had the highest sensitivity and specificity (83.6％ and 96.3％ respectively) for cholangiocarcinoma,and CA242 had a high sensitivity for gallstone canceration though it was not so sensitive in cholangiocarcinoma.Multiple tumor markers increased the sensitivity and specificity in the early diagnosis of cholangiocarcinoma.Compared with imageological determinations,multiple tumor markers had a higher sensitivity for the early diagnosis of cholangiocarcinoma with a tumor mass ＜1 cm.Conclusion Multiple tumor markers have important clinical values in the early diagnosis of cholangiocarcinoma.

Objective To explore the surgical management of chronic pancreatitis complicated with pancreatolithiasis (CPPL). Methods The clinical data of 66 patients with CPPL were retrospectively analyzed. Pancreatolithiasis was classified into 4 types according to the location of stones: stones located in the head of the pancreas (type Ⅰ, n=28), stones located in the body of the pancreas (type Ⅱ, n=30), stones located in the tail of the pancreas (type Ⅲ, n=1) and stones located from the head to tail of the main duct of pancreas (type Ⅳ, n=7). Ten patients (including 4 with type Ⅰpancreatolithiasis, 5 with type Ⅱ and 1 with type Ⅳ) received conservative treatment; 10 patients with type Ⅰ pancreatolithiasis underwent lithotomy under endoscope; pancreaticoduodenectomy and Beger procedure were carried out on 14 patients with type Ⅰ pancreatolithiasis, pancreatolithotomy+pancreaticojejunostomy on 25 patients with type Ⅱ pancreatolithiasis, resection of pancreatic tail and spleen on 1 patient with type Ⅲ pancreatolithiasis, and Puestow-Gillesby procedure, dividing of the neck of pancreas+removing stones from both ends of pancreatic duct+Roux-en-Y pancreatojejunostomy on 6 patients with type Ⅳ pancreatolithiasis. Results Sixty-two patients were followed up for 2 months to 15 years, and the number of patients with recurrence for type Ⅰ, Ⅱ, Ⅲ and Ⅳ pancreatolithiasis was 4, 2, 0 and 3, respectively. Conclusions Early surgical management according to the location of stones should be carried out after confirmed diagnosis of CPPL. Individualized management based on correct diagnosis and classification plays an important role in the prevention of pancreatolithiasis recurrence.

Objective To analyze the impact of electronic portal imagingdevice (EPID) position error on three-dimensional dose verification of volumetric modulated arc therapy (VMAT).Metbods Five Suremark SL-20 lead points were fixed on Elekta tray,and EPID images were collected in 0-360° rotation,one image per 5°.The position error relative to the accelerator was analyzed via Matlab.Then the images position error was corrected according to the analysis,and the 3D dose was reconstructed with the corrected images.The dose distributions of double arcs,clockwise arc(arc 1),and counterclockwise arc (arc 2) of 16 nasopharyngeal carcinoma patients' VMAT plan were evaluated by γ analysis,and the results of before and after position error correction were compared.Results Compared to 0° gantry angle,the error of source to the image distance (SID) was maximum (1.20 cm) when the gantry angle was 180°.On account of the SID change,the maximum error along the up-down (y) direction in the iso-center planar was 2.28 mm and the left-right (x) direction error was within ± O.5 mm.The 3D γ analyses of 16 nasopharyngeal carcinoma in VMAT plans were obviously increased after the position error along y was corrected.The double arcs,arc1 and arc 2 were increased by (4.12 ±1.67) ％ (t =-9.86,P＜ 0.05),(3.47±1.64) ％ (t=-8.46,P＜ 0.05) and (5.08±1.30) ％ (t=-15.63,P＜ 0.05) in 5％/3 mm standard,respectively.However,in 3％/3 mm standard,γ value of the double arcs,arc 1 and arc2 were increased by (7.63 ±2.24) ％ (t =-13.63,P＜ 0.05),(6.03 ±2.07) ％ (t =-11.66,P＜ 0.05),(9.17 ±2.23) ％ (t =-16.41,P＜ 0.05),respectively.Since the EPID position error along x was corrected after y,the 3D γ analysis of reconstruction dose indicated that the average of the 5％/3 mm and 3％/3 mm γ value were increased by 0.23％ and 0.24％,respectively.Conclusions EPID motion error along the gantry to table direction of the accelerator can't be ignored.When reconstruct dose based on EPID,a modification should be made for rebuilding more accurate patients' 3D dose distribution.