BACKGROUND: Studies have demonstrated that exogenous basic fibroblast growth factor (bFGF) has intensive effects to promote proliferation of gingival fibroblasts (GFs) cultured in vitro and the heeling of gingival wounds. OBJECTIVE: To investigate the effects of bFGF gene transfection on the biological performance of Beagle canine GFs. DESIGN, TIME AND SETTING: An observation and comparison in vitro experiment regarding cells was accomplished in Centre of Cell Biology and Development of Fujian Medical University and Department of Comparative Medicine in Fuzhou General Hospital of Nanjing Military Area Command of Chinese PLA from April to September of 2008. MATERIALS: Four beagle dogs, male, 12 months old, weighing 10-13 kg were used in this experiment, plRES2-EGFP-bFGF plasmid containing full-length human bFGF gene cDNA was constructed and conserved by our institution. METHODS: Free gingiva of the 2nd, 3rd and 4th premolars were excised from left upper jaw of Beagle dogs, dnsed with aseptic phosphate buffer four times, then cut into pieces and digested with 2.5 g/L pancreetin for 2 hours at 37 ℃. After the cantrifugation and supernatant removal, DMEM containing 10% fetal bovine serum was added to incubate on 6-well plate with coverlips in 5% CO2 incubator at 37 ℃. Logadthrnically growing cells were digested and passaged. GFs were transfected with pIRES2-EGFP-bFGF plasmid using liposome mediated method, while vacant plasmid transfection and un-transfection group served as controls.MAIN OUTCOME MEASURES: Proliferation and apoptosis feature of the GFs were evaluated by M'rE and AOEB, respectively. The activity of alkaline phosphatase was assayed by chemical coledmetry. RESULTS: All of three groups cells entered log phrase on three days after transfection. MTT results showed that the proliferation of GFs transfected with bFGF was greater than cells transfected with vacant vector and untransfected cells (P < 0.05). AO/EB dyeing showed the apoptosis rate of GFs transfected with bFGF was reduced compared with other two groups (P < 0.05). After bGFG gene transfection, the ALP activity remained unchanged and there was no significant difference compared with untransfected cells.CONCLUSION: The transfection of bFGF gene to GFs can promote the proliferation of GFs and depress the apoptosis. No promotion is present with regard to the GFs differentiation.

Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bleomycin ; administration & dosage ; analogs & derivatives ; Carcinoma, Squamous Cell ; drug therapy ; Cisplatin ; administration & dosage ; Combined Modality Therapy ; Drugs, Chinese Herbal ; therapeutic use ; Esophageal Neoplasms ; drug therapy ; Female ; Humans ; Male ; Middle Aged ; Mitomycin ; administration & dosage ; Phytotherapy

Background The development of retinopathy of prematurity(ROP) is associated with many regulatory cytokines related to neovascularization;however,the retinal expression and regulated mechanism of stromal cell-derived factor-1 (SDF-1) in mouse model of oxygen-induced retinopathy (OIR) remain uncertain.Objective This study was to investigate the expression of SDF-1 in retina of mouse model of OIR.Methods Forty 7-day-old C57BL/6J mice were divided into OIR group and control group.In OIR group,20 mice were exposed to 75% oxygen for 5 days and then to room air for 5 days.In control group,20 mice were raised in room air.The expression of SDF-1 in retina of mice was studied by immunochemistry and quantified by real time reverse transcriptase polymerase chain reaction (RT-PCR).Results The positive immunohistochemical staining for SDF-1 was found mainly locating at the ganglion cell layer in 12-day-old mice of OIR group;the stronger positive immunohistochemical staining for SDF-1 was noted mainly locating at the ganglion cell layer,vascular endothelial cells of inner retina,neovascular endothelial cells in 17-day-old mice of OIR group;the delicate positive immunohistochemical staining for SDF-1 was both found mainly locating at the inner retina and being around the retinal vascular in 12-day-old mice of control group and 17-day-old mice of control group.The expression of SDF-1 mRNA in 17-day-old mice of OIR group was higher than that of 12-day-old mice of OIR group (t=8.072,P＜0.05)and 17-day-old mice of control group(t=10.026,P＜0.05),respectively.The expression of SDF-1 mRNA in 12-day-old mice of OIR group was lower than that of 12-day-old mice of control group (t=4.336,P＜0.05).Conclusion SDF-1 might improve the onset of retinal neovascularization of OIR.

Objective To investigate the expression and significance of tumor necorisis factor related apoptosis induled ligand receptor(TRAILR) in human craniopharyngioma.Methods The expression of TRAILR was determined by immunohistochemistry and in situ hybridization in 24 samples of craniopharyngioma and 16 samples of normal brain tissue.Results With low decoy receptor(DcR) expression in partial craniopharyngioma cells and low death receptor(DR) expression in partial normal brain cells,DR was expressed highly in all craniopharyngioma samples while DcR in most normal brain tissue. High DR expression and low DcR expression in craniopharyngioma tissue differed from low DR expression and high DcR expression in normal brain tissue(P

Objective To study the expression level and clinical significance of bcl-XL gene in childhood medulloblastoma.Methods The expression of Bcl-XL protein and bcl-XL mRNA were determined by immunohistochemical staining and in situ hybridization in 41 samples of medulloblastoma tissues,as well as 20 normal brain tissues.Results The positive rate of Bcl-XL protein(90.2%) and bclXL mRNA(95.1%) in medulloblastoma group were significantly higher than those in normal human brain tissues(all P

OBJECTIVETo investigate the association of SOX9 expression and clinicopathologic factors and prognosis of gastric cancer.

METHODSA retrospective cohort study including 112 gastric cancer patients admitted to the Zhejiang Provincial People's Hospital from 2004 to 2006 was performed. Immunohistochemical analysis was used to evaluate the expression of SOX9 in the 112 specimens of gastric cancer tissues and 70 non-cancerous tissues adjacent to the tumor.

RESULTSLow expression of SOX9 was seen in 5(7.1%) tissues out of 70 non-cancerous tissues adjacent to the tumor. A total of 94(83.9%) patients had varying expression of SOX9, of whom 51(45.4%) had overexpression. Univariate analysis demonstrated that the expression of SOX9 was significantly associated with Lauren classification (P<0.05), tumor invasion(P<0.01), lymph node metastasis(P<0.05), distant metastasis(P<0.05) and tumor stage(P<0.05), however there was no significant association between SOX9 expression and sex, age, histological type, histology differentiation or tumor size. Kaplan-Meier analysis showed that the 5-year survival rate of patients with SOX9 over-expression was significantly lower than that of patients with low expression(29.4% vs. 49.2%, P=0.031). Multivariate Cox regression analysis showed that histology differentiation(P=0.046), tumor invasion(P=0.001), and distant metastasis(P<0.01) were independent prognostic factors for gastric cancer, however the over-expression of SOX9 was not significant(P=0.948).

CONCLUSIONSThe expression SOX9 is associated with the growth, invasion, and metastasis of gastric cancer, as well as the prognosis. However, SOX9 expression is not an independent factor for the prognosis in patients with gastric cancer.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; SOX9 Transcription Factor ; metabolism ; Stomach Neoplasms ; metabolism ; pathology

Objective To investigate the clinical characteristics and analyze prognostic risk factors of combined hepatocellular-cholangiocarcinoma. Methods The clinical data of 19 cases of combined hepatocellular-cholangiocarcinoma admitted in our hospital from January 1999 to December 2009 were analyzed retrospectively. The survival function was analyzed by Kaplan-Meier. The possible prognostic risk factors were tested by χ2-test. Results Hepatocellular-cholangiocarcinoma was diagnosed by pathology in the 19 patients, among which hepatic tunic was infiltrated in 13 cases, peritoneum involved in 1 case, intravascular cancer embolus in 1 case. At that time lymphocyte nodes metastasis in 2 cases were found by regional lymphadenectomy in 7cases. The 1-year and 3-year survival rates were 61% and 42%,respectively. Prognosis of patients with tumor size ＞ 5 cm ( χ2 = 4. 392, P = 0. 036 ), history of heavy drinking ( χ2 = 11.010, P = 0.001 ) or intraoperative blood transfusion ( χ2 = 4. 645,P = 0. 031 ) were worse than others. Conclusion It was difficult to get correct preoperative diagnosis of combined hepatocellularcholangiocarcinoma. Tumor size, history of heavy drinking and blood transfusion were all prognostic related risk factors.

Three-way laryngeal mask airway (tLMA) was used in 31 patients aged 4-68 yr, weighing 10- 79 kg undergoing tracheal foreign body removal under general anesthesia. Anesthesia was induced with propofol 3 mg/kg, vecuronium 0.12 mg/kg and remifentanil 0.4 μg/kg. tLMA was inserted. The patients were mechanically ventilated. Anesthesia was maintained with iv infusion of propofol 2 mg . Kg-1 ? H-1, vecuronium 0.08 mg·kg-1·h-1 and remifentanil 0.15 μg·kg-1 ·min-1 . Radial artery was cannulated for BP monitoring and blood sampling. The operation time was 6-34 min and mechanical ventilation time 19-45 min. There was no significant change in SP, DP, HR, VT, Ppeak and Ppeak CO, during operation as compared with the baseline values before anesthesia. SpO2 was significantly increased at T2-6. PCO2, PO2 and O2sat were obviously improved after tLMA was used. All the patients emerged bom anesthesia within 30 min after operation. No aspiration, obvious gastrointestinal inflation, and pharyngeal and laryngeal edema and injury occurred. Mild agitation occurred in a short time during the recovery period in one patient. No complication occurred.

OBJECTIVETo investigate the expression of matrix metalloproteinase-2 (MMP-2) and insulin-like growth factor-1 (IGF-1) in gastric carcinoma and their clinicopathological significance.

METHODSExpressions of MMP-2 and IGF-1 were examined by using immunohistochemical SP staining and cross-compared with clinicopathological features of gastric carcinoma.

RESULTSHigh expression of MMP-2 and IGF-1 were observed in 70.4% (307/436) and 49.5% (216/436) of gastric carcinoma tissues respectively, significantly higher than those in non-tumor gastric mucosa (3.3% and 5.4%, respectively; all P < 0.05). The high expression rate of MMP-2 and IGF-1 were significantly associated with the patient age, tumor size, tumor location, Lauren classification, TNM staging, depth of tumor infiltration, presence of vessel invasion, lymph node and distant metastasis (all P < 0.05). In addition, the expression of MMP-2 was positively linked with the expression level of IGF-1 (P < 0.05). Univariate analysis showed that high expression of MMP-2, was significantly associated with poor prognosis of tumor of TNM stage I and II (all P < 0.05), high expression of IGF-1 was significantly correlated with poor prognosis of patients with TNM stage I, II and III tumor (all P < 0.05). Cox multivariate analysis indicated that the high expressions of MMP-2 and IGF-1 could be independent prognostic indices for gastric carcinoma.

CONCLUSIONSHigh expression of MMP-2 and IGF-1 proteins are significantly correlated with the invasion and metastasis of gastric carcinoma, it is helpful to simultaneously detect the expressions of MMP-2 and IGF-1 proteins in predicting prognosis of patients with gastric carcinoma.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Insulin-Like Growth Factor I ; metabolism ; Lymphatic Metastasis ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Middle Aged ; Neoplasm Invasiveness ; Prognosis ; Stomach Neoplasms ; metabolism ; pathology ; Young Adult

OBJECTIVETo investigate the relationship between the expression of vascular endothelial growth factor (VEGF) and fms-like tyrosine kinase (Flt-1) mRNA and tumor progression, microvessel density and survival time in gastric carcinoma.

METHODSIn situ hybridization and immunohistochemical techniques were used to detect the gene expression of VEGF, Flt-1 and CD34 in 118 gastric carcinoma specimens.

RESULTSIn situ hybridization revealed that positive expression rates of VEGF and Flt-1 mRNA in gastric carcinoma were 54.24% and 55.9% respectively. There was a significant correlation between the expression of VEGF and Flt-1 mRNA and growth pattern, the depth of tumor invasion, vessel invasion, lymph node and distant metastasis (P < 0.01). The mean tumor microvessel densities (MVD) in patients of stage T3-T4 or those with vessel invasion, lymph node and distant metastases were significantly higher than those of stage T1-T2 and without metastases (P < 0.01). MVD value was correlated with the expression levels of VEGF and Flt-1 mRNA (P < 0.01). The mean survival time and survival rate of patients with positive mRNA expression and mean MVD value >or=54.9/mm2 were significantly lower than those of patients with negative mRNA expression and mean MVD value < 54.9/mm2.

CONCLUSIONSThe expression of VEGF and Flt-1 can promote tumor angiogenesis and contribute to tumor invasion and metastasis in gastric carcinoma. VEGF and Flt-1 may serve as valuable indicators of biological behaviour, prognosis and target of gene therapy in gastric carcinoma.

Adult ; Aged ; Female ; Humans ; In Situ Hybridization ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging ; Prognosis ; RNA, Messenger ; genetics ; Stomach Neoplasms ; metabolism ; pathology ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism