Objective To introduce the current studies of the role of vascular endothelial growth factor-C(VEGF-C) and VEGF-D in lymphangiogenesis and lymph node metastasis of gastrointestinal neoplasma.Methods The related literatures in recent 5 years were reviewed.Results The growth factors VEGF-C and VEGF-D enhance lymphangiogenic metastasis of gastrointestinal neoplasma with the property of angiogenesis and lymphangiogenesis.In gastric adenocarcinoma,VEGF-C mRNA and tissue protein expression correlate with lymphatic invasion,lymph node metastasis,venous invasion and reduced 5-year survival rates.The role of VEGF-C in esophageal squamous cancer and colorectal cancer and VEGF-D in colorectal cancer is not certain,with conflicting reports in the published literatures.Conclusion The VEGF-C,VEGF-D/VEGFR-3 signal pathway may become the ideal target for inhibition of tumor proliferation and metastases,antilymphangiogenesis therapy may be a novel potential strategy in tumor biological therapy.

BACKGROUND: Surface roughness of implants can directly influence cellular proliferation, differentiation, and gene expression. OBJECTIVE: To observe the early attachment of periodontal ligament cells (PDLCs) to pure titanium with different surface roughness levels, and to study the effect of surface performance on cell differentiation. DESIGN, TIME AND SETTING: Randomized controlled observation/multi-sample comparison study, which was performed at Center of Science and Technology, Gannan Medical College between January 2005 and July 2006. MATERIALS: Pure titanium stick was cut into pieces, of 10 mm diameter and 2 mm thickness, using cutting-off machine, and there were 24 sections in total. Then, the titanium sections were randomly divided into four groups: simple mechanical processing, nitric acid processing, sand blasting processing, and combination group, with 6 sections per group. METHODS: TR240 portable-type surface roughness meter was used in this study. In the simple mechanical processing group, sections were scoured by sand paper alone; in the nitric acid processing group, sections were etched with 65% HNO3 for 1 hour at 100 ℃ after scoured by sand paper; in the sand blasting processing group, sections were sandblasted by 100 μ m AI203 after scoured by sand paper; in the combination group, sections were etched with 65% HNO3 for I hour at 100～C after scoured by sand paper and sandblasted by 100 μ m Al2O3. MAIN OUTCOME MEASURES: Samples were maintained in DMEM for 30 minutes, and the third-passage cells were inoculated. Then, titanium sections were taken out at different time points of 30, 60, 120, 240 minutes, 1, 3, and 7 days. Surface roughness and early attachment of PDLCs were detected under fluorescent microscope. RESULTS: O Quantitative analysis: Surface roughness was (599.5±8.3) nm in the simple mechanical processing group, (406.5 +4.6) nm in the nitric acid processing group, (358.8±11.8) nm in the sand blasting processing group, and (8.7±2.0) nm in the combination group. On the other hand, surface roughness in the simple mechanical processing group was significantly higher Fluorescence observation exhibited that number of PDLCs attaching to pure titanium surface was increased, and the proliferation was greater with the time passing by. in addition, surface roughness of pure titanium was positively associated with number of PDLCs. CONCLUSION: The lighter the surface roughness is, the more the early attachment of PDLCs is, benefiting for cell adhesion and proliferation.

] Objective To explore the role and mechanisms of FGF2 in chemo?resistance in breast cancer. Methods The inhibitors for different signal pathway were used to treat two drug?resistant breast cancer cell lines MCF?7/5?Fu and T47D/5?Fu established in our lab. MTS assay was used to determine chemo?sensitivity and Hoechst stain was used to measure apoptosis. Protein activation and FGF2 protein level in cell culture medium were detected by western blot and ELISA respectively. Results Akt inhibitor MK?2206 (20 nM) and mTOR inhibitor AZD8055 (2 nM) significantly reversed the chemo?resistance of MCF?7/5?Fu and T47D/5?Fu cell lines to 5?Fu and paclitaxel, but ERK1/2 inhibitor SCH772984 showed no significant effect. Compared to parent cell lines MCF?7 and T47D, p?Akt and p?S6K (represented as mTORactivity) levels were obviously up?regulated in MCF?7/5?Fu and T47D/5?Fu cell lines, and so do the FGF2 mRNA level and FGF2 protein level from culture medium. Moreover, FGFR inhibitor AZD4547 (4 nM) markedly reversed the chemo?resistance of MCF?7/5?Fu and T47D/5?Fu cell lines to 5?Fu and paclitaxel and down?regulated activation of FGFR?Akt?mTOR signal pathway. In agreement, FGF2 protein (10ng/ml) enhanced the chemo?resistance of MCF?7 and T47D cell lines to 5?Fu and paclitaxel and up?regulated activation of FGFR?Akt?mTOR signal pathway. Conclusion Activation of FGF2?FGFR?Akt?mTOR signal pathway promoted chemo?resistance of breast cancer cells.

Objective To explore the role of insulin-like growth factor-2/insulin-like growth factor1 receptor/insulin receptor substrate-1 (IGF2/IGF1R/IRS1) signal pathway inducing the chemoresistance of epidermal growth factor receptor 2 (ErbB2) positive breast cancer cells to Herceptin.Methods Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot assay were used to determine the expression levels of IGF2,IGF1 R,and IRS1.The direct targets of miR-126 were validated by dual-luciferase reporter gene assay.In SKBR3/pool2 cells,IGF1 R activity was reduced by an inhibitor of IGF1 R,and IRS1 was knocked-down by shRNAs.Furthermore,3-(4,5-dimenthylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was performed to evaluate the sensitivity of these treated cells to Herceptin.Results IGF2,IGF1 R,and IRS1 were significantly higher expressed in SKBR3/pool2 cell compared to that in SKBR3 cell.Western blot assay showed that IGF2/IGF1R/IRS1 was activated in SKBR3/pool2 cells.Bioinformatics analysis combined with luciferase activity suggested that miR-126 directly targeted IRS1.MTS results demonstrated that the chemosensitivity to Herceptin of SKBR3/ pool2 cells with inhibitor of IGF1R or shRNAs targeting IRS1 or overexpressing miR-126 was significantly reduced.Conclusions IGF2/IGF1R/IRS1 signal pathway confers to the chemoresistance of ErbB2 positive breast cancer cells to Herceptin.

Objective To investigate the expression of PCNA in gastric cancer and its relationship with telomerase activity of peritoneal washings and peritoneal dissemination, and to compare the efficacy of telomerase activity and cytology of peritoneal washings for prediction of peritoneal metastasis of gastric cancer. Methods Telomeric repeated amplification protocol (TRAP)-enzyme-linked immunosorbent assay (ELISA) was performed to measure the telomerase activity of peritoneal washings collected from 60 patients with gastric cancer. Exfoliate cytologic analysis of the corresponding samples was used for comparison.Expression of PCNA was measured with immunohistochemical staining.Their relationship with clinicopathologic features were evaluated. Results The positive rate of telomerase activity in peritoneal washing collected from patients with gastric cancer was 41.7%,which well related to serosal invasion, histology types, depth of infiltration and peritoneal metastasis of gastric cancer. The positive rate of telomerase activity increased with the increased depth of infiltration and serosal involvement areas (P

Objective: To evaluate the clinical significance of serum fibronectin and prealbumin as nutritional parameters in nutrition support after abdominal surgery.Methods: Serum albumin,transferrin,prealbumin and fibronectin were determined respectively before and 7 days after nutrition support in abdominal surgery patients,and nitrogen balance was calculated daily during nutrition support.Results: Compared with before nutrition support,the levels of serum transferrin,prealbumin and fibronectin were significantly increased after nutrition support.Furthermore,the change of prealbumin was correlated with that of fibronectin.Conclusion: Measurement of serum transferrin and prealbumin can be recommended as valuable indicators in assessing the nutritional status of surgery patients receiving nutritional support.

Objective:To investigate the clinical significance of dynamic analysis of plasma brain natriuretic peptide in patients with dilated cardiomyopathy with chronic heart failure.Methods:Ninety patients with dilated cardiomyopathy with chronic heart failure admitted into our hospital firom March 2012 to March 2016 were divided into group A (20 cases),group B (38 cases),and group C (32 cases) according to the NYHA grading.The plasma BNP levels and LVEF,LA,LVEDD,and LVESD in the three groups were detected and compared.The correlation of plasma BNP and cardiac function and ultrasonic cardiogram indexes were analyzed.And the capability of plasma BNP and LVEF in diagnosis of patients were analyzed and compared.Results:The plasma BNP level in group C was markedly higher than that of group A and group B (P＜0.05),and that in group B was much higher than that of group A (P＜0.05).And LA in group C was significantly higher than that of group A (P＜0.05),while differences in LVEF,LVEDD,and LVESD were not obvious (P＞0.05).The plasma BNP was positively correlated to NYHA grading,but had no significant correlation with the LVEF,LVEDD,LVESD,and LA (P＞0.05).Based on results of receiver operating characteristic curve analysis,plasma BNP =523.5 pg/mL was the threshold value for identification of patients with NYHA Ⅲ and Ⅳ (AUC=0.901,P＜0.001),while LVEF had not the capability (AUC=0.392,P=0.276).Conclusion:Detection of plasma BNP level had important clinical significance on diagnosis,screening and cardiac functional grading of patients with dilated cardiomyopathy with chronic heart failure.

BACKGROUND: The investigation of culturing, passaging and establishing human limbal stem cells can strengthen the recognition of the stem cells and provide the enough cellular reserve for the basic and clinical research of limbal stem cell transplantation.OBJECTIVE: To explore a method of pessaging and establishing cell line of human limbal stem cells cultured in vitro.DESIGN: Randomized controlled observation.SETTING: Gannan Medical College.MATERIALS: The experiment was performed at Scientific Center of Gannan Medical College and the National Key Laboratory of Ophthalmology Hospital Affiliated to Sun Yat-Sen University from June 2003 to April 2004. Fresh human limbus corneae were isolated from two healthy donors. Procedures were performed according to the informed consent of the donors. Main reagents contained RPMI-1640 (Sigma R8755, containing L-glutamine) and 200 g/L fetal calf serum (FCS) (Gibco 16140-071). DMEM medium, chondroitin sulfatase and human epidermal growth factor (hEGF) were purchased from Sigma Co. USA; HEPES and DMSO were bought from Gibco, USA; 100% glycerinum was purchased from Yunjia Huangpu Pharmaceutical Product Limited Company, PRC; glutaraldehyde was bought from E.Merk, Germany; Alcohol, chlorhydric acid, acetone and methyl aldehyde were purchased from Beijing Chemical Agent Company, PRC; 0.25% parenzyme was bought from Shanghai Xinhua Pharmaceutical Factory, PRC.Above-mentioned reagents were analytical pure grade.METHODS: After digestion, human limbal tissues in limbal basilar part with an abundant pigment were cultured in the culture flask containing RPMI-1640 and 200 g/L FCS and in culture dish containing amniotic extracellular matrix (AECM) as the cultural supporter. Primary and passage cells were observed under light microscope and scanning electron microscope (SEM). The revival ratio of stem cell refrigeration of every generation was calculated by the trypanblau exclusion experiment.MAIN OUTCOME MEASURES: ① Observational results of limbal stem cells during the primary culture and serial subcultivation in vitro, and ② revival ratio of stem cell refrigeration.RESULTS: ①Findings of primary culture: Most limbal stem cells in the culture flask had the adherence and were arrayed uniformly sparsely to form monolayer and adhered to the bottom of culture flask under the inverted phase contrast microscope after 1-day culture. ② Findings of serial subcultivation: After human epidermal growth factor (hEGF) was added into the second passage, cells were scattered into the monolayer and adhered to grow quickly.Morphological variability of all the cells increased obviously when passage the 30th generation. The cellular volume was obviously increasing, and the round or irregular round cells gathered together. The 33rd generation human limbal stem cells still could vigorously differentiate, proliferate and grow in ACEM. ③ The revival ratio of stem cell refrigeration was 82.2%.CONCLUSION: The human limbal stem cell lines were preliminarily established by culturing and freezing the cells of 33 generations in vitro. The human limbal stem cell lines preferred to grow in the culture dish containing AECM as the cultural supporter.

BACKGROUND: Stromal cell-derived factor 1 (SDF1), a potential inhibitor of infection by lymphophilic HIV-1 strains, can help to block the pathway of HIV-1 invasion into the human body.OBJECTIVE: Genotype and polymorphism of SDF1-3 'A allele associated with HIV-1 infection were investigated in She Ethnic Group in the south of China so as explore the possible causes of uninfection by HIV-1 strains among this population.DESIGN: Single sample study.SETTING: Department of Biochemistry and Molecular Biology, Gannan Medical College.PARTICIPANTS: Totally 186 She Ethnic subjects without HIV-1 infection collected randomly from those whose three generations belonged to She Ethnic Group, and inhabited in Qianshan County of Jiangxi Province,Ningde area of Fujian Province and Jingning She County of Zhejiang Province, from January to December 1995.METHODS: The whole blood samples from 186 She Ethnic subjects were collected randomly, and then their genomic DNA samples were extracted respectively. Allelic polymorphism was examined by the polymerase chain reaction and restriction-fragment-length polymorphisms (PCR-RFLP).MAIN OUTCOME MEASURES: The distribution of SDF1-3'A allele in She Ethnic Group in the south of China.RESULTS: The data of 186 She Ethnic subjects entered the result analysis without any loss in the midway. The frequency of SDF1-3 'A allele in She Ethnic Group samples was 19.6%, and the allelic distribution of the gene was in accordance with Hardy-Weinberg equilibrium. No difference was found between male and female individuals.CONCLUSION: The frequency of SDF1-3 'A allele of She Ethnic Group in the south of China was similar to that of Dai Nationality in Yunnan.Based on its slow-down effect on clinical course of AIDS, the mutation of SDF1-3'A is significant in the prevention and treatment of AIDS in She Ethnic Group in the south of China.

ObjectiveTo investigate the characteristics of EGFR gene mutations among NSCLC patients in south of China and analyze the correlation between mutations and clinical features.Methods Specimens of lung cancer tissues were collected from 185 NSCIC patients in our hospital.DNA was extracted from specimens.Exon 18,19,20 and 21 of EGFR gene were amplified by FQ-PCR to be bi-directional sequenced.ResultsEGFR gene mutations in 62 (33.5％) of 185 NSCLC patients was identified in carcinoma tissues,of which,2cases,41cases,5 cases and 14 cases respectively located at exon 18,exon 19,exon 20 and exon 21.The mutation of Del L747 → P752 (P753S) ( proportion 8.1％ ),Del E746 → A750 ( proportion 45.1％ ) at exon 19 and L858R ( proportion 22.6％ ) at exon 21 were the predominant mutation in 16 kinds of mutations.Four cases of mutation at exon 19 got the different results in bi-directional sequencing.The silent mutation 2361G→A at exon 20 was observed (28.1％ ).The mutation rate in women was significantly higher than men (46.2％ vs 24.3％,x2 =9.670,P =0.002).Non-smokers had significantly higher mutation rate than smokers (41.4％ vs 17.1％,x2 =7.380,P =0.007) ; Adenocarcinoma patients had significantly higher mutation rate than squamous cell carcinoma (38.3％ vs 6.3％,x2 =6.426,P =0.011).Clinical stage Ⅲ patients had significantly lower mutation ratethan patients with stage Ⅱ orⅣ ( 10.8％ vs 53.8％,x2 =8.026,P =0.003 ;10.8％ vs 41.3％,x2 =9.518,P =0.002).No statistically significance correlation was found between the mutation ratio and age.ConclusionsEGFR gene mutation has a close relationship with females,non-smokers and adenocarcinoma.Most mutations occur in exon 19 and 20 among patients in south of China.