Since its first application for radical gastrectomy, laparoscopic surgical technique has been used for almost 20 years. Howev-er, given the complexity of the anatomic plane and vessels of stomach and the particular operation of laparoscopic instruments, lapa-roscopic lymph node dissection is still a difficult technique that requires experience. An appropriate approach is the key point to the success of laparoscopic lymph node dissection. On the basis of lymphatic drainage routes and anatomy of perigastric lymphatic sys-tem, different approaches are selected to dissect periodic gastric lymph nodes safely and radically. Surgery is performed according to the patient's position, surgeons' stance, and sequence of lymphadenectomy for laparoscopic gastrectomy and lymph node areas, such as suprapancreatic, cardial, and splenic hilar areas. In this article, we introduce different surgical approaches of laparoscopic radical gastrectomy with extended lymphadenectomy to provide information for surgeons who want to perform this surgery.

82 spastic cerebral palsy and normal children were selected, including 27 younger and 27 older spastic cerebral palsy( observation group) ,and 14 younger and older normal children( control group) . The tumor necrosis factor α( TNF-α) levels were measured in the serum with ELISA. The TNF-α levels in the serum of the younger and older spastic CP groups were higher than those of the control groups ( P<0.01 ) . In observation groups, the TNF-α levels of younger spastic cerebral palsy were higher than those of the older (P<0.01). On the contrary, the normal children in the control group of different ages had no difference of TNF-α. TNF-αexpression was signifi-cantly higher in the serum of the spastic CP group than that in the control group ( P<0.01 ) . TNF-α expressed continuously in vivo of children with spastic cerebral palsy involve subsequent brain injury.

BACKGROUND: It is important to study the methods of culturing bone marrow-derived mesenchymal stem cells (MSCs) to obtain a great amount of high purity MSCs for applying ocular tissues constructed by tissue engineering technique to treat eye diseases.OBJECTTVE: To separate and culture in vitro MSCs from bone marrow of the adult rats by density gradient centrifugation combined with adherence culture, and observe the growing characteristics and the possibility of mass multiplication.DESIGN: A completely randomized grouping design/repetitive measuring experiment.SETTING: Pathological laboratory, Zhongshan Ophthalmic Center, Sun Yat-sen University.MATERIALS: Four six-week-old SD rats about 250 g, grade Ⅱ of cleaning, were provided by the Animal Center of Sun Yat-sen University [certificate number: SCSK(Yue)2004/0011], about 250 g each rat and there was no limit to the sex. The main reagents and instruments included low sugar Dulbecco modified Eagle culture medium (DMEM/F12, American Gibco Corporation), trypsin (fetal bovine serum (FBS, Hangzhou Sijiqing Bio-Engineering Material Research Institute), American Gibco Corporation), disodium edetate, lymphocyte separating medium, fibronectin, CD44, CD34, CD31 monoclonal antibodies, two-step-method kit for immunohistochemistry (Beijing Zhong shan Biotechnology Corporation).METHODS: This experiment was conducted at the Key laboratory of Ophthalmology (Sun Yat-sen University), Ministry of ethanol (750 g/L) for 10 minutes. Under aseptic condition, the medullaris cavitas was exposed, the syringe containing application m edium was directly punctured into the femoral cavity, the cells in the medullaris cavitas were washed out with the culture medium containing heparin and taken as the cell suspension. The bone marrow-derived MSCs were separated and purified by density gradient centrifugation combined with adherence culture, and the growing conditions of the wells. When the cells had generally connected with each other, they were fixed with methanol or dimethy ketone in situ for 10 minutes, and then hematoxylin eosin (HE) staining or immunohistochemical staining. Antibodies against fiinoculated to 96-well culture plate by a cell density of 4.25×107/L, with 200 μL every well; then put the culture plate into culture box. Then from the next day to the sixth day, 5 g/L MTT solution was added into two rows (20 μL every well) every day, continuously cultured for 4 hours, then the supernatant was removed, and 200 μL DMSO was added to each well, agitated for 5 minutes, then detected the absorbance (A) values at 570 nm wave length, and a growth curve was drawn.branes were clear and the cell bodies were lucent. Being cultured for 2 days, there appeared adherent cells. 3 days later,most of the adherent cells extended and appeared to be in polygon, star or long-shuttle shapes. 4 days later, the cells showed to be in division growth stage; and about 12 days later, the cell clones were connected to each other, appearing curve of subcultured MSCs was in S shape. Cells began growing fast on the 2nd day after being passaged, and they entered the growth period for 3-4 days followed by saturation period, and then cells stopped growing.CONCLUSION: It is a simple and practical method to separate MSCs from the bone marrow of adult rats by means of density gradient centrifugation and adherence. MSCs can greatly proliferate in vitro and offer seed cells for the application of tissue engineering technique to treat eye diseases.

Background The pathological basis of human primary open angle glaucoma mainly is the degeneration of trabecular meshwork and Schlemm canal.As the outflow pathway of aqueous humor, the tissue origin and characteristics of trabecular meshwork remain less clear.Studying the characteristics of trabecular meshwork may contribute a new thought to the treatment of primary open angle glaucoma.Objective This study was to explore the histological property of trabecular meshwork by detecting the expression of D2-40, a lymphatic biomarker,and CD31,CD34 and smooth muscle actin (SMA), some vascular endothelial biomarkers.Methods Twenty specimens of adult eyeballs were collected from Zhongshan Ophthalmic Center of Sun Yat-sen University,including l0 eyeballs from accident death donors,3 enucleated eyeballs due to posterior segment of choroidal malignant melanoma and 7 orbital exenterated eyeballs.The series ocular meridian sections with the thickness of 4 μm were prepared for the haematoxylin-eosin staining.Immunohistochemistry was employed to detect the expression of D2-40,CD31 ,CD34 and SMA in trabecular meshwork and Schlemm canal.Results Intact structure of chamber angle was observed under the optical microscope by haematoxylin-eosin staining.Human trabecular meshwork tissue was presented with meshlike structure,while Schlemm canal showed the lumen structure.No abnormal substance and periphery synechia were found.D2-40 was strongly expressed in the trabecular meshwork endothelial cells with staining score for 3 points, but CD31, CD34 and SMA were negatively expressed.In contrast, CD31, CD34 and SMA rather than D2-40 were positively expressed in the Schlemm canal.In addition, CD31 and CD34 were also positively expressed in the vascular endothelial cells around the trabecular meshwork.Conclusions The trabecular meshwork expresses lymphatic biomarker rather than vascular biomarkers.This result indicates that trabecular meshwork of human eyes is lymphatic vessel-like tissue.

ObjectiveTo investigate the efficacy of laparoscopic spleen-preserving hilar lymph nodes dissection based on splenic hilar vascular anatomy.MethodsFrom July 2010 to March 2011,the clinical data of 39 patients with advanced proximal gastric cancer who underwent laparoscopic spleen-preserving hilar lymph nodes dissection at the Union Hospital of Fujian Medical University were retrospectively analyzed.Different types of vascular anatomy were analyzed,and different methods of lymph node dissection in the splenic hilus were adopted accordingly.ResultsThe operation was successfully performed on all the patients,with no conversion to open surgery or splenectomy due to splenic vascular or parenchyma injury.There were 4 types of splenic artery running,including type Ⅰ (25 patients),type Ⅱ (8 patients),type Ⅲ (4 patients) and type Ⅳ (2 patients).There were 2 types of the end branches of splenic artery,including concentrated type (28 patients) and dispersion type (11 patients).The splenic lobial vessels of all the patients were anatomically classified and divided into 4 types,including a single branch of splenic lobial vessels in 3 patients,2 branches in 24 patients,3 branches in 11 patients and multibranches in 1 patient.The mean number of short gastric vessels was 3.2 ± 1.4 (range,2-6).The time for dissection of the lymph nodes in the splenic hilum,number of lymph nodes dissected in the splenic hilum,volume of operative blood loss,duration of hospital stay and incidence of complications were ( 30 ±7)minutes,2.8 ±2.1,(20 ±7)ml (range,0-55 ml),(10 ± 1) days and 10％ (4/39).All patients were followed up until March,2012. One patient had hepatic metastasis,and no patient died postoperatively.ConclusionFamiliar with the variation of splenic hllar vascular anatomy is helpful in mastering and promoting laparoscopic spleen-preserving hilar lymph nodes dissection.

Objective　To evaluate the expression of Fas/FasL antigen on peripheral blood T lymphocytes and its possible role in Behcet disease. Methods　The expression of Fas and FasL antigen on peripheral blood T lymphocytes was determined by flow cytometry and three-colour double marked immunofluorescence methods in 26 patients with Behcet disease and 43 healthy individuals. Results　The difference was significant between Behcet disease (25.70%±7.32%) and controls (14.02%±6.30%) concerning the Fas expression on CD4+ T lymphocytes(P＜0.01) . But no difference was found concerning the expression of FasL antigen between Behcet disease and controls (P＞0.05).The expression of Fas antigen on CD8+ T lymphocytes from Behcet disease (9.47%±6.97%)was significantly higher than that in control group(3.47%±2.75%), but no difference was found concerning FasL antigen expression on CD8+ T lymphocytes between Behcet disease and controls(P＞0.05).Conclusion　These results indicate the imbalance of the expression of Fas and FasL on T lymphocytes in Behcet disease is responsible for the perpetuation and recurrence of Behcet disease for the activated lymphocytes would not be eliminated through apoptosis mediated by Fas/FasL system.

Splenic hilar lymph node (LN) is the difficulty and focus of perigastric LN dissection during radical gastrectomy for advanced proximal gastric cancer (APGC). Although the new edition of Japanese Gastric Cancer Treatment Guidelines excluded the splenic hilar LN from the D₂ lymphadenectomy in patients with APGC, there are still few high-level evidences. Meanwhile, the guideline recommends that splenectomy is still necessary for patients with APGC involving the greater curvature. With the support of increasing evidences, the spleen-preserving splenic hilar LN dissection has been recognized by more and more scholars. Additionally, the technique of laparoscopy provides a new method for splenic hilar LN dissection. Our center proposed a new technique named Huang′s three-step maneuver for laparoscopic spleen-preserving splenic hilar lymphadenectomy, which not only reduces the difficulty of operation, but also makes the technique easier to popularize and with a significance of minimally invasive effect. During the clinical practice, the dissection of LNs behind the splenic portal vessels is also one of the controversies. Therefore, the difficult technique of splenic hilar LN dissection not only requires us to make a technical exploration, but also requires us to explore precise indications, in order to avoid unnecessary operation in patients who do not require dissection of the No.10 LNs and to avoid missing the opportunity for radical surgery in high-risk patients because of the difficult techniques.

Objective To investigate the impact of preoperative comorbidities on the abdominal complications after laparoscope-assisted total gastrectomy (LATG) for gastric cancer.Methods The retrospective casecontrol study was conducted.The clinical data of 1 657 gastric cancer patients who underwent LATG at the Fujian Medical University Union Hospital between January 2008 and December 2015 were collected.There were 175 patients with postoperative abdominal complications,including 78 without preoperative comorbidities and 97 with preoperative comorbidities (52 with 1 comorbidity,30 with 2 comorbidities and 15 with more than 3 comorbidities).Analysis method and observation indicators:(1) risk factors analysis of abdominal complications after LATG;(2) risk assessment of abdominal complications after LATG:independent influencing factors of risk factors analysis were expressed as dependent variables,alignment diagram was built and then consistency index was calculated;(3) comparisons of abdominal complications among the patients with different kinds of comorbidities after LATG;(4) multivariate analysis of abdominal complications in patients with comorbidities after LATG;(5)follow-up situations.Follow-up using outpatient examination and telephone interview was performed to detect postoperative survival of patients up to May 2016.The univariate analysis and multivariate analysis were respectively done using the chi-square test and Logistic regression model.The survival rate was calculated by the Kaplan-Meier method.Results (1) Risk factors analysis of abdominal complications after LATG:results of univariate analysis showed that age,body mass index (BMI),number of preoperative comorbidities,operation time and estimated volume of intraoperative blood loss were related factors affecting abdominal complications of patients after LATG (X2 =4.487,16.602,10.361,4.567,7.482,P＜0.05).Results of multivariate analysis showed that BMI,number of preoperative comorbidities and estimated volume of intraoperative blood loss were independent factors affecting abdominal complications of patients after LATG [OR =1.966,1.204,1.423,95％confidence interval (CI):1.355-2.851,1.014-1.431,1.013-1.999,P＜0.05].(2) Risk assessment of abdominal complications after LATG:BMI,number of preoperative comorbidities and estimated volume of intraoperative blood loss were expressed as dependent variables,and the alignment diagram on risk prediction of abdominal complications after LATG was built,with a consistency index of 0.703.(3) Comparisons of abdominal complications among the patients with different kinds of comorbidities after LATG:numbers of patients without comorbidity,with 1 comorbidity,2 comorbidities and ≥3 comorbidities were detected in 21,8,13,3 patients with intra-abdominal infection and 13,10,9,5 patients with anastomotic leakage and 6,3,6,2 patients with intra-abdominal bleeding,respectively,with statistically significant differences (X2 =10.677,10.436,9.245,P＜0.05).(4) Multivariate analysis of abdominal complications in patients with comorbidities after LATG:BMI ≥25 kg/m2 and estimated volume of intraoperative blood loss ＞ 82 mL were independent risk factors affecting abdominal complications of patients with preoperative comorbidities after LATG (OR =2.104,1.771,95％ CI:1.307-3.387,1.146-2.738,P＜0.05).(5) Follow-up situations:of 1 657 patients,1 568 were followed up for 4-99 months,with a median time of 47 months.Ninety-seven patients with preoperative comorbidities undergoing LATG had postoperative abdominal complications and were followed up.During follow-up,5-year survival rate of patients was 58.1％,and 5-year survival rate of 97 patients with preoperative comorbidities undergoing LATG and with postoperative abdominal complications was 57.4％.Conclusion Preoperative comorbidities are independent factors affecting abdominal complications of patients after LATG.

AIM:To isolate and culture tumor stem cells in human retinoblastomas (RTSC). METHODS: Retinoblastoma (RB) single cells acquired from fresh tumors of RB patients by enzyme digestion were seeded in serum-free medium at a density of 1?10~8 cells/L. Clonal cultures were plated at a density of 1?10~6 cells/L. Secondary tumor spheres were triturated again and passaged in fresh medium. The sphere-forming, proliferation and differentiation assays were performed. RT-PCR and immunocytochemistry were performed to identify the RTSC and differentiated cells. RESULTS: All RB tumors studied produced proliferating neurosphere-like tumor spheres, which were also passaged multiple times. These tumor spheres had the capability to self-renew, proliferate in SFM medium, expressed retinal progenitor cell related genes, and differentiated into neurons and glia when they were transferred to differentiation conditions.CONCLUSION:Our findings demonstrated that there were subsets of tumor stem cells resembling retinal progenitor cells in human RB, which can be isolated, cultured in SFM. The RTSC may be original cells of RB tumor, and also become the new target of tumor therapy.

Recently, the technological advancements have led to the introduction of new three-dimension (3D) laparoscopic system. 3D technique in laparoscopic radical gastrectomy provides an abdominal strong stereoscopic depth and accurate operative field, and can be helpful for the beginner to shorten the learning curve. Moreover, spatial orientation becomes more accurate with 3D technique. 3D technique also has the advantages of clearer anatomical level, better vessel exposure, lymph node dissection and digestive tract reconstruction. However, there are still some defects of 3D laparoscopic system. For example, the surgeon may not adapt to 3D vision at the beginning, and special equipment is required to display the data.
Gastrectomy ; methods ; Humans ; Imaging, Three-Dimensional ; methods ; Laparoscopy ; Learning Curve ; Lymph Node Excision ; Reconstructive Surgical Procedures ; Stomach Neoplasms ; surgery