Objective: To establish analysis method of vascular endothelial growth Factor(VEGF)for clinic practice.Methods: the expression of VEGF in breast carcinoma tissue is examined with immunohistochemistry, RT-PCR, Western blot and dot blot. Results: the VEGF is revealed mainly in tumor cells and extracellular matrix, molecular weight of VEGF is 46 KD, including three isoforms. In 53% of the cases the tumor tissue reveals stronger expression of VEGF than paratumor tissue. Conclusion :Dot blot may be used as a preliminary method for detecting VEGF in breast carcinoma tissue.

Objective To explore the aberrant expressions of mammalian target of rapamycin (mTOR) and its activation form,pmTOR,in stage IIIB human colon cancer,and analyze the association of mTOR and pmTOR with clinicopathologic characteristics and patients' prognosis. Methods A total of 104 patients with stage IIIB (T 3-4 N1M0) colon cancer with available follow-up data were enrolled. The specimens of colon cancer and clinical data were acquired from these subjects. The expressions of mTOR and pmTOR in tumor tissues and matched normal tissues were detected with immunohistochemistry. The relationship and its significance between the expressions of mTOR and pmTOR and some clinicopathological features,such as gender,age,location of primary tumor,pathological stage and TNM stage,were analyzed. Results The mTOR and pmTOR were found to be expressed in cell membrane and cytoplasm of tumor tissues. Out of the total 104 specimens,positive rate of mTOR and pmTOR accounted for 75.9% (79/104) and 76.9% (80/104),respectively. No significant correlation was noted between the expressions of mTOR and pmTOR and the clinicopathological features by partial correlation analysis. Both univariate analysis and multivariate analysis showed that the expressions of mTOR and pmTOR threw no significant influence on the 5-year overall survival of patients. Nevertheless,subgroup analysis suggested that a high expression of pmTOR along tumor border was likely to contribute to higher risk of death in patients. Conclusion Aberrant expressions of mTOR and pmTOR are noted in patients with locally advanced colon cancer. High expression of pmTOR along the border of tumor tissue may suggest the possibility of invasion and metastasis.

Background and purpose:Dendritic cells(DCs) possess specialized feature such as pathogen recognition,antigen capturing and processing machinery,and stimulating naive T lymphocyte to have antitumor ability that allow them to act like professional APCs.This paper is aimed to confirm the impacts on the proliferation and secretion of INF-? of tumor specific CTL and its cytotoxocity induced by DC loaded with different antigen.Methods:After the stimulation of DC loaded with different antigen,the proliferative rate of allolymphocytes was measured by MTT and the cytotoxocity of CTL was evaluated with LDH method.The INF-? secreted by activated T lymphocytes was detected by ELISPOT.Results:The DC loaded with CPP-Id(320%?15%) had significantly induce T lymphocyte proliferating when comparing with the induction by DC loaded with Id(57%?10%)(P

Objective:To examine the oncogenic mutations involved in melanoma in Southern China and to provide a theoretical basis for the development of melanoma molecular targeted therapy strategy. Methods:The Sequenom platform (OncoCarta Panel v1.0 and MassARRAY System) was used to determine the prevalence of oncogene mutations in 28 acral melanoma samples, 28 mucosal mel-anoma samples, and 30 non-chronic sun-induced-damage (no-CSD) melanoma samples from Southern China. Results:At least one mu-tation was detected in 33 of the 86 melanomas (38.4%) with mutations observed in BRAF (16.3%), NRAS (10.5%), KIT (5.8%), EGFR (4.7%), HRAS (2.3%), KRAS (2.3%), MET (2.3%), and PIK3CA (1.2%). In BRAF, the age of patients with mutations was significantly lower than those without BRAF mutation (45.7±15.3 vs. 55.9±12.7, P=0.01). Patients with mutations in NRAS were more likely to have ulceration compared with patients without NRAS mutations (88.9%vs. 48.1%, P=0.049). Conclusions:This study represents a compre-hensive and concurrent analysis of the major recurrent oncogenic mutations involved in melanoma cases from Southern China areas. The data have implications for both clinical trial designs and therapeutic strategies.

Objective: To observe the tolerability of Chinese melanoma patients to four-week high-dose interferon alfa-2b(INTRON A(R)，Schering-Plough)therapy. Methods:A total of 29 patients with high risk melanoma[American Joint Committee on Cancer Staging(AJCC)ⅡB-ⅢC]who received adjuvant interferon therapy in our hospital between September 2007 and May 2009 were retrospectively reviewed.Patients received 4 hours of intravenous infusion of interferon alfa-2b fdose range，22.00 million international unit(MIU)to 33.75 MIU]Ⅳ 5 days/week for 4 weeks.The adverse events were evaluated with National Cancer Institute Common Toxicity Criteria(NCI 2.0 version). Results: The average daily dose was 17.63 MIU/(m~2·d).The therapy was ended in two patients because of poor wound healing or intolerability to severe fatigue.The most common adverse events were myelosuppression.Grade 3/4 neutropenia was observed in 69% (20/29)patients and was rapidly reversed after conventional support interventions.Grade 1/2 abnormal hepatic function occurred in 18 cases(62%).Twenty-six patients were followed up for 3 to 22 months.Five patients developed early progression:one with local recurrence，two with regional lymph node metastasis one with in-transit metastasis in the affected limb,and one with distant metastasis. Conclusion: High-dose interferon alfa-2b regimen can be well tolerated by Chinese patients but cannot effectively inhibit subclinical lesions.

Objective:To investigate the labial(buccal)bone thickness at alveolar crest zone and alveolar crest height of the maxil-lary anterior teeth and premolars of young adults with normal occlusion.Methods:The alveolar bone of the anterior teeth and premo-lars of 67 eligible Han national young volunteers was scanned by CBCT.Then the facial bone thickness and the distance between the facial alveolar crest and Cemento-enamel Junction(CEJ)of the anterior teeth and premolars were measured and analyzed after recon-struction.Results:The distance between labial (buccal)crest and labial (buccal)CEJ of the maxillary first premolars was the lar-gest(P <0.05);there was a negative correlation between the labial(buccal)crest height and the facial alveolar bone thickness at 2 mm from CEJ toward root derection[(P <0.05),0.6 <|r|<0.8].Conclusion:The labial (buccal)crest of the first premolars was higher than that of other teeth in maxillary aesthetic zone.At alveolar crest zone,when the labial (buccal)bone was thinner,the dis-tance between labial (buccal)crest and labial (buccal)CEJ was larger,and the implant aesthetic risk is higher.

[Objective]To analyze the prognostic factors of resectable acral melanoma patients ,then develop a novel prognostic model and examined its prognostic value.[Methods]The study retrospectively analyzed clinicopathological characteristics and inflam?matory markers of 232 acral melanoma patients who underwent radical surgical resection between 2000 and 2011 at the Sun Yat-sen University Cancer Center. Kaplan-Meier curves were plotted to estimate overall survival. Significantly predictive factors were identified by multivariate Cox regression analyses and a prognostic model based on these variables was constructed to predict survival.[Results]Cox regression analysis revealed that age,lactic dehydrogenase(LDH),stage,globulin(GLB)and C-reactive protein (CRP)were independently related to survival. After computing these scores ,patients were classified into three risk groups. The new prognostic model identified three categories of patients with different prognoses(P<0.001)and significantly stratify patient prognosis in different tumor stages. The 5-year survival rate was 42.9%,25.7%,and 3.7%in groups 1,2,and 3,respectively. The AUC of new prognostic model is 0.664(95%CI:0.599-0.724).[Conclusion]Age,LDH,stage,GLB and CRP were independently related to survival in our study population,and the prognostic model is useful to stratify patients into different risk groups and it is a useful complement to AJCC staging for Asian patients with acral melanoma.

Objective To extract,isolate and identify the polysaccharide from ginseng,and to investigate its anti-tumor activity in vitro. Methods The ginseng polysaccharide was obtained through water extraction and ethyl alcohol deposition method. Use the Sevage method to remove the protein in the crude polysaccharide. The structural characteristics of the polysaccharide were determined by FT-IR spectra.The RM-1 and HeLa cells were divided into control group and different concentrations (0,50,100,200,300,400 and 500 mg·L-1 )of ginseng polysaccharide groups. The survival rates of the cells in various groups were detected by MTT method. The indirect killing effects of ginseng polysaccharide with different concentrations (0,50,100,200,300,400 and 500 mg·L-1 )on the cancer cells were determined by CTL test. Results The extraction rate of ginseng polysaccharide was 8.7 6% and the structural characteristics demonstrated that the main component of the extract was polysaccharide.The result of MTT showed that there were no significant differences of the survival rates of RM-1 and HeLa cells between different concentrations of ginseng polysaccharide groups after treated for 24 h compared with control group (P>0.05).The result of CTL test showed that the cytototic LHD release rates in different concentrations of ginseng polysaccharide groups were increased significantly (P<0.05)compared with control group;with the increase of ginseng polysaccharide concentration, the cytotoxic LDH release rates were increased firstly and then were decreased.When the concentration of ginseng polysaccharide was 100 mg·L-1 ,the cytotoxic LDH release rate was the biggest.Conclusion Ginseng polysaccharide can indirectly inhibit the growth of the tumor cells by activating T cells and play an anti-tumor effect.

Objective To investigate the expression of platelet derived growth factor(PDGF) in small bowel transplantation of rats.Methods Isogeneic and allogeneic small bowel transplantation were performed in rats by microsurgical technology.All rats were divided into two groups:isogeneic control group and allogeneic test group.Transplanted tissues were test on 7th,28th and 90th after surgery.Positioning using immunohistochemical method the expression of PDGF.Real time PCR and immunohistochemical staining were also performed to detect the expression of PDGF.Results The unique feature including intestinal graft fibrosis was showed in tissues.Immunohistochemistry results showed PDGF expression was higher in intestinal glands.PDGF mRNA levels in transplanted tissues showed a gradual upward trend,and the top levels is in POD90.Conclusion PDGF expression was significantly higher in the late of intestinal transplantation,which showed an guideline for chronic rejection of intestinal transplantation.

AIM:To evaluate the correlation between microRNA-1284 (miR-1284) and gastric cancer, and to investigate the underlying mechanism.METHODS: The expression of miR-1284 was examined by real-time PCR in 63 gastric cancer ( GC) tissue samples and 63 non-malignant adjacent tissue samples.The correlation between miR-1284 and the clinicopathological feature of GC was analyzed.Lentiviral vector containing miR-1284 was constructed and transfected into GC SGC-7901 cells.After transfection, the expression of miR-1284 was examined by real-time PCR.The cell activity was evaluated by CCK-8 assay.The cell cycle and apoptosis were determined by flow cytometry.The ability of cell migra-tion was measured by wound-healing assay.The potential target gene of miR-1284 was predicted by online bioinformatic softwares.The expression of JAG1 mRNA was examined by real-time PCR.The protein levels of JAG1, Notch1 and NF-κB were analyzed by Western blotting.RESULTS:Compared with non-malignant adjacent tissue samples, the results of real-time PCR showed significant downregulation of miR-1284 in 42 GC tissue samples ( P<0.05 ) .The expression level of miR-1284 was not significantly associated with age and gender of the patients, tumor size, TNM staging and lymph node metastases (P>0.05), but significantly associated with histologic grading (P<0.05).Compared with LV-NC-GFP group and control group, after transfection of miR-1284 in LV-miR-1284 group, the expression of miR-1284 was significantly in-creased (P<0.05), the percentages of apoptotic cells and the cells in G0/G1 phase were significantly increased (P<0.05), the cells activity and ability of migration were significantly decreased (P<0.05), and the expression of JAG1, Notch1 and NF-κB was significantly decreased (P<0.05).CONCLUSION:The inhibitory effect of miR-1284 on gastric cancer may be associated with the regulation of its targeting gene JAG1.