Digestive tract reconstruction is the main part of gastrointestinal surgery. With the rapid development of technology and widely application in stapling device, more and more surgeons are using stapled anastomosis. Stapled anastomosis is associated with shorter operating time and hospital stay than hand-sewn anastomosis. However, it is not easy to select suitable ones from various staplers and use them correctly. Choice and reasonable application of staplers for anastomosis in gastrointestinal surgery are summarized and evaluated in this article.
Anastomosis, Surgical ; instrumentation ; Digestive System Surgical Procedures ; instrumentation ; Humans ; Surgical Staplers

Objective: To construct the recombinant lentiviral vector expressing specific shRNA of p120 catenin gene, and to identify the RNA interference efficiency by infecting PANC-1 cells. Methods: Three pairs of shRNA targeting p120ctn mRNA were designed, synthesized and ligated into enzyme-digested and linearized pGCSIL-GFP lentiviral vectors. The recombinant lentiviral vectors were co-transfected with the pGC-FU-p120ctn-3FLAG containing p120ctn gene into 293T cells after identification by PCR and sequencing analysis. Western blotting analysis was applied to select the most effective shRNA. Recombined lentivirus was packaged and the concentration of the virus titer was measured. Real-time PCR and Western blotting analysis were used to identify the interference efficiency after infection of the PANC-1 cells by recombined lentivirus. Results: PCR and DNA sequencing analysis confirmed that p120ctn-shRNA-LV was successfully constructed and the concentration of the virus titer was 3×109TU/ml. Real-time PCR showed that expression of p120ctn mRNA was decreased by 82.6% in PANC-1 cells infected with the recombined lentivirus (P<0.05). Western blotting analysis showed that the expression of p120ctn protein was also greatly deceased after infection. Conclusion: We have successfully constructed lentiviral vector expressing shRNA of p120 catenin gene, and this lays a foundation for studying the role of p120ctn in invasion and metastasis of pancreatic carcinoma.

Hepatic artery stricture (HAS) after liver transplantation can lead directly to transplanted liver function exhaustion and complications of biliary system. The early diagnosis and treatment are crucial for better prognosis. Doppler ultrasound is the first method of choice, and angiography can give further clear dignosis. The balloon dilatation is still effective for hepatic arterial stenosis. With the more adaptable usage of oronary stent, if possible, would reveal more promising result especially for tortuous stenotic hepatic artery. The vascular reconstruction or repeated liver transplantation is still the effective therapeutic methods.

China ; Ethnic Groups ; genetics ; Female ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; Receptors, Complement ; genetics

Objective To explore the change characteristics of physiological indexes between infants and children during living-donor liver transplantation and discuss methods of regulation and control.Method In this study,42 patients were selected and assigned into two groups according to age:infants group (＜1 year,n =25),and children group (1-16 years,n =17).The preoperative and peri-operative characteristics,intra-operative operation conditions,internal environment changes before and after re-perfusion,postoperative mechanical ventilation time,ICU time,hospital time,infection rate,additional surgery,complications and survival were analyzed.Result PELD (MELD) score,historical surgery rate and hematokrit were lower in children group than in infants group (P＜ 0.05).Serum creatinine and lactate concentrations increased significantly in children group as compared with infants group (P＜0.05).Intra-operative an-hepatic phase and cold ischemia time were shortened significantly (P ＜ 0.05),and incidence rate of re-perfusion syndrome was reduced in children group as compared with infants group (P＜0.05).As compared with pre-re-perfusion,blood lactate concentrations were significantly raised only in infants group and glucose concentrations significantly raised only in children group (P＜0.05).The blood levels of K + were decreased after reperfusion in both two groups,and those in infants group were lower than in children group (P＜ 0.05).Postoperative intensive care unit time was longer in children group than in infants group (P＜ 0.05),and there was no significant difference in survival rate between two groups.Conclusion There are many differences and change characteristics to physiological indexes between infants and children during the operation of living-donor liver transplantation.Timely management and regulation are critical for the success of surgery according to the differences.

Aim Toinvestigatetheeffectofrecombi-nant snake venom metalloproteinase inhibitor (rSVM-PI ) on neovascularization and its molecular mecha-nism.Methods Chickenchorioallantoicmembrane (CAM)assay was used to examine the antiangiogenic effect of rSVMPI.Alamar blue analysis was used to de-tect cell proliferation.Annexin V-FITC double labeling flow cytometry was used to assay cell apoptosis. Scratch marker was used to assay cell migration.Boy-den chamber analysis method was used to detect cells chemotaxis in vitro.Tube like structure(TLS)of HU-VECs was used to detect the ability of neovasculariza-tion in vitro.Real-time PCR and Western blot were used to assay the expressions of KDR and FGFR-1 inHUVECs. Results Thevasculardensityindex (VDI)of CAM was drastically decreased after rSVMPI treatment, chemotaxis of HUVECs in response of VEGF was inhibited in the presence of rSVMPI,TLS of HUVECs was less than control group.The expres-sions of KDR and FGFR-1 were down-regulated by re-al-timePCRandWesternblotassay.Conclusion rS-VMPI may inhibit neovascularization by blocking the VEGF-KDR or bFGF-FGFR signal transduction path-way.

Background Retinal development continues during the early postnatal period in mammals.Correct arrangement of layers and precise location of various cells in the retina are vital for forming normal visual function during critical period plasticity.Spectral-domain optical coherence tomography(SD-OCT)provides highquality in vivo retinal imaging and the possibility to measure retinal thickness longitudinally. Objective The present study was to investigate the changes of retinal thickness during critical period plasticity in rats. Methods In vivo consecutive scanning of retinal image was performed in 10 SPF Sprague-Dawley rats at postnatal day 14(P14),P18,P21,P24 and P42 with SD-OCT,and retinal histopathological examination was used to detect retinal morphologic changes at the same postnatal ages in 20 matched rats.The whole retinal thickness,the thickness from inner limiting membrane(ILM)to inner plexiform layer(IPL),the thickness of inner nuclear layer(INL)and the thickness from outer nuclear layer(ONL)to retinal pigment epithelium(RPE)were measured using Cirrus HD-OCT system and HMIAS-2000 Imaging System in retinal sections.The measurement parameters by Cirrus HD-OCT and those by hematoxylin-eosin staining were compared.The use of animals followed the Statement of National Institute of Health (USA). Results In vivo high-resolution images of rat retinas with SD-OCT compared well with histology,which enabled quantitative comparison of the SD-OCT and histological data during critical period plasticity in rats.From P14 to P42,the retinal thickness gradually decreased with the increase of rat ages(F=15.425,P=0.000),and so were the thickness from ILM to IPL,the thickness of INL and the thickness from ONL to RPE(F=3.973,P=0.007;F=17.529,P=0.000;F=7.038,P=0.000).The retinal thickness,thickness of INL.thickness from ONL to RPE measured by Cirrus HD-OCT were significantly correlated with those measured by retinal sections among P14,P18,P21,P24 and P42 rats(r=0.794,P=0.000;r=0.784,P=0.000;r=0.681,P=0.000). Conclusion SD-OCT is a demonstratably valuable technology to study the structure of retinas in rats.The retinal thickness is shown to reduce in thickness throughout the development of the retina during critical period plasticity due to the decrease in thickness of INL and the distance from the ONL to RPE,as illustrated by OCT scanning.

Background To optimize the culture method of human retinal microvascular endothelial cells is very important for the study of retinal angiogenesis disease.Human retinal microvascular endothelial cells have been successfully cultured in previous studies,but further improvement of the culture method to harvest higher yields and purity cells is still needed.Objective This study was to design a modified method to isolate and purify human retinal microvascular endothelial cells much easily and quickly,and to compare the expression of specific markers of vascular endothelial cells,factor Ⅷ and CD31/CD34 in the cells.Methods The use of human donor eyeballs was approved by the Ethic Commission of Zhongshan Ophthalmic Center of Sun Yat-sen University.The retina tissue from healthy donor was isolated and digested by the two-step digestion method with 2％ trypsin and 0.133％ collagenase Ⅳ.Human retinal microvascular endothelial cells were collected and plated in 60 mm dishes coated by 0.1％ fibronectin and cultured in endothelial cell-specialized medium supplemented with 10％ fetal bovine serum,0.3 mg/L β-endothelial cell growth factor (ECGF) and 100 ng/L sodium heparin.During the culturing,the growth situation of the cells was monitored by morphological observation,and immunohistochemical staining was performed to probe vascular endothelial cell-specific membrane protein CD31,CD34 and factor Ⅷ for identification of the cell purity.Results Human retinal microvascular endothelial cells were isolated successfully from the retina by the twostep digestion method.The primary cultured cells adhered to well 72 hours later and achieved confluence with the typical cobblestone appearance 9 to 10 days after cultured.The cells exhibited the blue nuclei and reddish cytoplasm by regular haematoxylin and eosin stain and showed a strong positive response for CD31,CD34 and factor Ⅷ by immunohistochemistry.The positive dye of CD31 and CD34 was lower than Ⅷ factor in both endothelial cells.Conclusions Modified culture method of human retinal microvascular endothelial cells can improve cell culture result and purify target cells.

Objective To identify risk factors associated with the severity of acute kidney injury (AKI) in-duced by crush syndrome and whether the patient required hemodialysis (HD). Method A retrospective study was designed. Within 19 days after the Chinese Wenchuan earthquake (May 12, 2008), 63 victims (33 men, 30women) of 2139 cases were hospitalized at Mianyang Central Hospital (Siehuau, China) because of crush syn-drome caused by crush injuries. The patients with renal dysfimcfion before the earthquake were excluded. Totally 63 patients with AKI associated with crush syndrome were included in this study and were divided into two groups: group 1, 25 patients, requiring HD (when urine volume <250 mL/d;serum potassium> 6 mmol/L) ; and group 2, 38 patients, without HD. The following data were collected retrospectively for all patients: (1) epidemiological parameters: age, gender, race, time under the rubble, liquid treatmem before being rescued; (2) clinical param-eters: blood pressure, body area crushed, amputation, fasciotomy, blood transfusion, quantity of fluid infusion, urine output in the first 24 hours; (3) initial laboratory data: complete blood count, urine analysis, serum chem-istry, arterial blood gas analysis. Comparisons between the two groups were made using SPSS 10.0. The quantita-tive data and categorical data were analyzed using t tests and χ2 tests, respectively. P -values < 0.05 were consid-ered to indicate significant differences. The significant variables were entered into logistic regression models to de-termine the risk factors for the severity of AK1 in patients with crush syndrome and whether the patient required HD. Results Four significant risk factors with P -values < 0.05 were identified: fasciotomy, cystatin C (Cys C)level, myoglobin (MB) level and lactic acidosis. The odds ratios (95% confidence intervals) were 8.641 (3.027～76.479), 6.956 (3.027～76.479), 5.379 (3.027～76.479) and 4.833 (2.569～32.764), respectively. Conclusions In addition to urine output and potassium levels, we found that four risk factors, namely faseiotomy, Cys C and MB levels, and lactic acidosis, were significanfly associated with the severity of AKI and whether the patient required hemodialysis.

Objective To investigate the effects of CO_2 pneumoperitoneum on the expression of focal adhesion kinase (FAK) of gastric cancer MKN-45 cells. Methods CO_2 pneumoperitoneum with different pressures was simulated in vitro, and the gastric cancer MKN-45 cells were divided into test and control groups. In the test group, gastric cancer MKN-45 cells were cultured in CO_2 pneumoperitoneum with different pressures [5, 10 or 15 mm Hg (1 mm Hg =0.133 kPa)] for 4 hours. The condition of the cells exposed to CO_2 pneumoperitoneum with a pressure of 15 mm Hg was observed at 0.5, 2 and 4 hours. Gastric cancer MKN-45 cells in control group were cultured at normal atmospheric pressure. The expression of FAK and phosphorylated FAK (FAK Tyr397) of each group was detected by Western blot. Multiple-group analysis was done by one-way ANOVA, and intergroup comparison was done by LSD test. Results In CO_2 pneumoperitoneum with pressures of 5, 10, 15 mm Hg, the expression of FAK was 2.14±0.17, 2.07±0.21 and 2.52±0.26, respectively, and the expression of FAK Tyr397 was 1.82±0.28, 1.93±0.52 and 3.71±0.37, respectively. The expression of FAK and FAK Tyr397 in the control group was 2.43±0.46 and 1.71±0.23, respectively. We found significant differences between the 2 groups (F = 2.171, 26.951, P < 0.01). After gastric cancer MKN-45 cells being treated for 0.5, 2 and 4 hours in CO_2 pneumoperitoneum with a pressure of 15 mm Hg, the expression of FAK Tyr397 was 3.41±0.44, 4.12±0.56 and 5.24±0.41 respectively, which is also significantly different (F =116.119, P < 0.01). The expression of FAK Tyr397 was back to 0.72±0.16 1 hour after the release of CO_2. Conclusions CO_2 pneumoperitoneum with different pressures can not promote the expression of FAK in gastric cancer MKN-45 cells which had been cultured for 4 hours, but can activate FAK through promoting its phosphorylation. The degree of FAK phosphorylation increases with pressure and time, and the activity of FAK decreases to pretreatment level rapidly once pressure is released.