Objective To investigate the effects of lymphangiogenesis on cancer growth and metastasis.Methods Human lymphatic endothelial cells(HLyECs)were isolated and purified from human lymph node by magnetic beads coated with antibody against human VEGFR3.Human breast cancer cell line(MDA-MB-435)or human osteosarcoma cell line(MG-63)was inoculated alone or co-inoculated with HLyECs subcutaneously into nude mice.The weight of tumor and lung surface metastasis were detected;peri-tumor lymphatic vessels were shown by Evans blue,intra-tumor lymphatic vessels were shown by immunohistochemistry for human and mouse PDPN.Conditioned medium of tumor on proliferation of HLyECs was evaluated by MTT assay.Results Compared with inoculating tumor cells alone,the growth and metastasis of MDA-MB-435 were promoted by co-inoculating HLyECs.The peritumoral and intra-tumoral lymphatic vessel density of MDA-MB-435 were increased by co-inoculating HLyECs.Both hPDPN-and mPDPN-positive lymphatic vessels were found.But co-inoculating HLyECs had no effect on lymphatic vessels density of MG-63.Neither intra-nor peri-tumor lymphatic vessels were found.The proliferation of HLyECs was increased by conditioned medium of MDA-MB-435 but not by MG-63.Conclusion Growth and lymphangiogenesis of breast cancer can be enhanced by co-inoculating lymphatic endothelial cells.

Objective To investigate the effect of IH764-3 on the leukocyte-mediated hypoxia-reoxygention injury of human brain microvascular endothelial cell (HBM VEC). Methods MTT assay was used to detect the survival of HBMVEC; gelatin zymography was used to check the activity of MMPs. The level of reactive oxygen species (ROS) in leukocyte was determined via commercially available kit, and the indirect enzyme-linked immunosorbent assay (ELISA) was used to quantify the contents of TNF-α, IL-1α, IL-2 and INF-γ in leukocyte culture medium. Results Survival of HBMVEC was impaired by hypoxia-reoxygenation, which was aggravated by supernatant of activated leukocytes but was attenuated by IH764-3. Leukocytes produced high level of MMP-9, ROS and cytokines (TNF-α, IL-1α, IL-2, IFN-γ) after hypoxia-reoxygenation, the process was inhibited by IH 764-3. Furthermore, IH764-3 could effectively reverse hypoxia-reoxygenation injury of HBMVEC with supernatant of activated leukocytes. Conclusion IH764-3 can protect HBMVEC from leukocyte-involved hypoxia-reoxygenation injury by attenuating the activation of leukocytes and inhibiting the pathogenic effects of leukocytes products.

In INS-1 cells, the insulin secretion was investigated by radioimmunoassay (RIA) after 4 h incubation in medium containing different concentrations of glucose and recombinant human glucagon-like peptide-1 (rhGLP-1) (7-36). Insulin mRNA level in INS-1 cells was assessed by a semi-quantitative RT-PCR method. rhGLP-1 (7-36) is not only a powerful insulin secretagogue, but also can increase insulin gene expression in INS-1 cells.

Objective To investigate the effect of IH764-3 on the leukocyte-mediated hypoxia-reoxygention injury of human brain microvascular endothelial cell (HBMVEC). Methods MTT assay was used to detect the survival of HBMVEC; gelatin zymography was used to check the activity of MMPs. The level of reactive oxygen species (ROS) in leukocyte was determined via commercially available kit,and the indirect enzyme-linked immunosorbent assay (ELISA) was used to quantify the contents of TNF-?,IL-1?,IL-2 and INF-? in leukocyte culture medium.Results Survival of HBMVEC was impaired by hypoxia-reoxygenation,which was aggravated by supernatant of activated leukocytes but was attenuated by IH764-3. Leukocytes produced high level of MMP-9,ROS and cytokines (TNF-?,IL-1?,IL-2,IFN-?) after hypoxia-reoxygenation,the process was inhibited by IH 764-3. Furthermore,IH764-3 could effectively reverse hypoxia-reoxygenation injury of HBMVEC with supernatant of activated leukocytes. Conclusion IH764-3 can protect HBMVEC from leukocyte-involved hypoxia-reoxygenation injury by attenuating the activation of leukocytes and inhibiting the pathogenic effects of leukocytes products.

Objective: To establish an in vitro model of brain blood barrier (BBB) using cultured mouse brain microvascular endothelial cells (BMVEC). Methods: Mouse BMVEC were seeded on micro pore membrane of gelatin coated cell culture insert and cultured to confluence. The establishment of BBB was preliminary judged by a 4 h water leaking test. The tight junctions between BMVEC were demonstrated by scanning and transmission electron microscope. The transendothelial electrical resistance(TEER) over BMVEC was measured. The permeability of Horseradish peroxidase (HRP) through the BBB was analyzed and the effect of RMP 7 on permeability of the BBB was investigated. Results: The 4 h water leaking test became positive when BMVEC were cultured to confluence. By scanning and transmission electron microscope, the tight junctions were demonstrated on confluent BMVEC. The TEER over BMVEC monolayer increased 3.2 and 7.68 times and the permeability rates for HRP were 13.4% and 6.7% respectively, as compared with sub confluent BMVEC and human umbilical vein endothelial cell monolayer(HUVEC). The HRP permeability rate in the model of BBB increased 2.7 times after treatment with RMP 7. Conclusion: The established in vitro model of BBB has basic characteristics of BBB in vivo , and is suitable for central nervous system (CNS) drug research over BBB.

OBJECTIVETo review the clinical features of duodenal gastrointestinal stromal tumors(GISTs), and to compare the clinical efficacy among different surgical treatments for duodenal GISTs.

METHODSClinicalpathological data of 36 cases of duodenal GISTs undergoing operation in The First Affiliated Hospital of Sun Yat-sen University from January 2000 to July 2013 were retrospectively analyzed. All the patients received surgical treatments, including 15 cases with regional resection, 8 cases with segmental resection, 12 cases with pancreaticoduodenectomy (PD), and 1 case with liver biopsy, respectively. Clinical efficacy between pancreaticoduodenectomy (PD) and non-PD (NPD) was compared.

RESULTSNine of 36 cases (25%) developed postoperative complications who were all in the PD group. Eight patients recovered and healed finally after active treatment, and 1 case was complicated with acute pancreatitis, pancreatic fistula and intra-abdominal infection. The median follow-up time was 54 months and the 5-year overall survival (OS) rate and 5-year recurrence-free survival (RFS) rate were 78.1% and 72.1%, respectively. The 5-year OS rate in the PD group and the NPD group was 61.1% and 61.1% respectively. The 5-year RFS rate in the PD group and the NPD group was 85.8% and 78.8% respectively. Statistical analysis showed no significant difference between the both groups (P=0.71 and P=0.89).

CONCLUSIONSFor duodenal GISTs patients, regional resection and segmental resection have similar clinical outcomes to pancreaticoduodenectomy while the former two can obviously decrease the incidence of postoperative complications. Based on the premise of R0 resection guaranteed, regional sectional and segmental resection with less injury should be the surgical treatment of choice.

Duodenal Neoplasms ; Gastrointestinal Stromal Tumors ; Humans ; Intraabdominal Infections ; Pancreatic Fistula ; Pancreaticoduodenectomy ; Postoperative Complications ; Retrospective Studies ; Survival Rate

OBJECTIVETo investigate the clinicopathological features of gastric cancer in Southern China, and provide a base of research and therapy for gastric cancer.

METHODSA total of 1 879 cases of gastric cancer with radical gastrectomy from Southern China were collected from August 1994 to July 2012. Analyze and summarize the characters of gender, age, tumor location, WHO histopathologic type and grade, pTNM stage and family history, retrospectively.

RESULTSAmong all cases, male to female ratio was 2.08: 1, while female was more than male before 40 years (χ(2) = 77.831, P = 0.000). Cases aged over 60 years had a highest incidence of gastric cancer (46.0%), with predilection of sinus (45.7%), body (26.3%) and cardia (20.1%). The common WHO histopathologic types were tubular or papillary adenocarcinoma (81.5%) and signet ring cell carcinoma (11.0%). Most patients were at III or IV stage on pTNM staging (40.5% and 26.5%).

CONCLUSIONSGastric cancer in Southern China has a predilection on male, while female is more than male before 40 years. Gastric sinus and adenocarcinoma is most common. Most patients are diagnosed at advanced stages.

Adult ; Aged ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Retrospective Studies ; Stomach Neoplasms ; epidemiology ; pathology ; Young Adult

OBJECTIVETo establish subcutaneous xenograft models of gastric cancer in nude mice and to screen the predictive biomarkers of bevacizumab effectiveness.

METHODSSubcutaneous xenograft models were established using BGC823 gastric cancer cell line in 20 male 4-week old BALB/C-nu/nu nude mice and were randomly divided into four groups, bevacizumab group(15 mg/kg), 5-FU group(15 mg/kg), combined group and control group, with 5 mice in each group. Bevacizumab and 5-FU were administered intraperitoneally every other day for three weeks. After treatment, tumor size and inhibition rate were calculated. Expression of CD31 was examined by immunohistochemistry for evaluation of microvascular density(MVD). Levels of human vascular endothelial growth factor(VEGF), basic fibroblast growth factor (bFGF), placental growth factor (PIGF) and interleukin 8(IL-8) were tested by enzyme linked immunosorbent assay(ELISA).

RESULTSCompared to the control group, bevacizumab group and combined group had a significantly lower MVD(5.2±1.0 and 4.3±1.2 vs. 13.8±1.6, P<0.05), a smaller tumor volume [(305.6±184.1) mm(3) and (242.2±71.4) mm(3) vs.(1535.2±625.1) mm(3), P<0.05], and lower levels of VEGF and IL-8 in tumor tissues [VEGF:(351.6±84.1) ng/L and (242.2±71.4) ng/L vs. (1256.7±702.1) ng/L, P<0.05); IL-8:(20 903±1485) ng/L and (27 489±6772) ng/L vs. (57 032±2437) ng/L, P<0.05]. The above parameters were not significantly different between 5-FU group and control group(all P>0.05). Levels of bFGF and IGF were not significantly different among four groups as well(all P>0.05).

CONCLUSIONVEGF and IL-8 may be used to be biomarkers candidates to predict bevacizumab effectiveness on human gastric cancer.

Animals ; Antibodies, Monoclonal, Humanized ; Bevacizumab ; Biomarkers ; Cell Line, Tumor ; Fluorouracil ; Heterografts ; Humans ; Immunohistochemistry ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Stomach Neoplasms ; Vascular Endothelial Growth Factor A ; Xenograft Model Antitumor Assays

BACKGROUNDAs a common form of gastric cancer migration, lymph node metastasis largely affects the surgical treatment and prognosis of gastric cancer. Surgery is the fundamental curative option for gastric cancer that varies depending on different stages. The study aimed to compare the clinicopathological characteristics and lymph node metastatic patterns in patients of proximal gastric cancer with different T stages and investigate a reasonable radical gastrectomy approach in terms of the range of lymphadenectomy for proximal gastric cancer.

METHODSIn our retrospective study, the data of 328 patients of proximal gastric cancer with different T stages were analyzed. By comparing the differences of lymph node metastatic rate and ratio, we investigated the clinicopathological characteristics and metastatic patterns of lymph nodes. Also, we were especially interested in the differences in survival rates between patients with and without No. 5 and 6 group metastasis with the same TNM stage.

RESULTSThe overall lymph node metastatic rate and ratio of advanced proximal gastric cancer were 73.4% and 23.3%, respectively. The tumors of different T stages were statistically significant in size and differentiation degree (P < 0.05), multivariate analysis showed that the depth of tumor invasion was an independent risk factor for lymph node metastasis in proximal gastric cancer (RR, 12.025; 95% CI, 2.326 to 62.157; P = 0.003). The overall survival rate of patients with No. 5, 6 group lymph node metastasis and those without was significantly different, but the differences in survival rates between patients with and without No. 5 and 6 group metastasis with the same TNM stage were not statistically significant.

CONCLUSIONSDifferent T stages in proximal gastric cancer showed different patterns and characteristics of lymph node metastasis. D2 lymphadenectomy in patients with early gastric cancer had little survival benefit because metastasis to level 2 nodes was rare. Therefore the range of the lymph node dissection in radical gastrectomy for early gastric cancer was considered reasonable. Moreover, to meet the requirements of the lymph node dissection, total gastrectomy plus D2 lymphadenectomy or more are supposed to be applied for the advanced proximal gastric cancer patients. Precise T staging largely determines the range of gastrectomy and lymphadenectomy.

Aged ; Female ; Gastrectomy ; Humans ; Lymph Node Excision ; Lymphatic Metastasis ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Stomach Neoplasms ; pathology ; surgery