Objective To study the effect of gastrin on the proliferation and angiogenesis of human umbilical vascular endothelial (HUVE) cell in vitro.Methods The immunocytochemistry assay,realtime-PCR,and Western blot were used to detect the gastrin receptor (CCK-BR) expression in HUVE cells.After HUVE cells were treated with 10 and 100 nmol/L gastrin for 72 h,MTT and soft agar colony formation assay were used to test the cell proliferation rate and colony formation rate,and the vascular-like structures were observed by three-dimensional culture of HUVE cells.The half-ring and ring vascular numbers were counted with five random visions.The vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1a (HIF-1a) mRNA and protein levels in HUVE cells were detected by realtime PCR and ELISA assay respectively.Results HUVE cells expressed CCK-BR.After the treatment with 10 and 100 nmol/L gastrin,the cell proliferation rate was increased by 48.48％ and 82.82 ％ compared to the control group,and colony formation rate was increased to 31.33％ and 45.67 ％ as compared with 15.33％ in the control group(P＜0.01).Relative expression quantities of VEGF and HIF-1a genes were 2.3 and 4.6 folds (VEGF) and 20.76 and 26.77 folds (HIF-1 a) than those in the control group.The concentration of VEGF protein in culture medium was 221 and 392 μg/mg protein higher than that in control group.The numbers of half-ring and ring vascular structures were (14.00 ± 3.00,39.33 ± 7.57 and 34.33 ± 4.50)/vision and (8.33 ± 2.51,41.33 ± 5.85 and 37.67 ± 3.51)/vision in control,10 and 100 nmol/L gastrin-treated groups,respectively (P＜0.01).Conclusion Gastrin up-regulates the expression of VEGF and HIF-1a genes in HUVE cells and promotes cell proliferation and vascular-like structure formation of HUVE cells in vitro by being combined to CCK-BR,which may be involved in the development and metastasis of gastric cancer.

Objective To study whether Helicobacter pylori CagA protein can control gastrin gene expression and the detailed mechanism. Methods First, pcDNA3. 1ZEO (-)/cagA7 was transfected into gastric cancer cell lines AGS and SGC-7901 cells. At the same time, culturing the Helicobacter pylori NCTC11637 and infecting AGS and SGC-7901 cells with it. Next, in the infected and transfecled AGS and SGC-7901 cells, respectively adding the JAK2 signaling pathway inhibitor AG490 and the ERK signaling pathway inhibitor U0126 to inhibit the two signaling pathway. Untreated gastric cancer cells and empty vector transfected cells as the control. Using real-time fluorescence quantitative PCR to detect the levels of gastrin mRNA in transfected and infected cells. Results After AGS and SGC-7901 cells were transfected with pcDNA3. lZE0(-)/cagA7 and infected with NCTC11637, the results showed that the expression of gastrin mRNA increased significantly (P ＜ 0. 05) in transfected and infected cells as compared with the control group, but after adding the inhibitor AG490 and U0126 respectively, the expression of gastrin mRNA decreased significantly(P＜0.05). Conclution These results suggest that CagA may up-regulate the expression of the gastrin gene, and CagA is one of the important proteins in regulating gastrin gene expression. The ERK/MAPK and JAK/STAT signaling pathways may be involved in controlling of gastrin gene expression by CagA.

Objective To investigate the correlation between food intolerance and Henoch-Sch(o)nlein purpura (HSP) in children and the efficacy of food forbidden or alternative therapy. Methods The levels of IgG against several common food in serum obtained from 40 children with HSP were measured by ELISA. The efficacy of food forbidden or alternative food therapy was assessed after 3 months. Results Total positive rate of serum food-intolerant IgG antibodies in HSP children was 92.5%. Among these 14 intolerant foods, the positive percentage of egg was the highest (33.8%), followed by tomato (14.9%), milk (13.5%) and morrhua (12.2%). Significant differences of the sort and degree of food intolerance existed among different age groups (t=2.257, P=0.045), but not between boys and girls (t=1.053, P=0.315), city and countryside (t= 1.388, P=0.193). There was no linear correlation between total food intolerance and serum IgG level (t=0.793, P=0.445). Food intolerance had no direct relation to immune complex deposition in kidneys of HSP nephritis (r =-0.262, P =0.387). The efficacy of adjusting diet was 95% . Conclusions HSP is closely related to food intolerance. Egg is the most common intolerant food. Food forbidden or alternative therapy shows acceptable efficacy in the treatment of most Henoch-Sch(o)nlein purpura children.

Objective:To discuss the application of antithrombotic drugs in myocardial infarction patients with thrombocytopenia. Methods:The therapy for one myocardial infarction patient with primary thrombocytopenia was discussed and summarized. Based on the related literatures, the characteristics of case, application status and adverse reactions of antithrombotic drugs and anticoagulant drugs were summarized. Results: The adjustment of antithrombotic drug treatment regimen for the patient was reasonable. Conclu-sion:Antithrombotic drugs applied in acute myocardial infarction patients with thrombocytopenia are safe. If patients complicate with other diseases, individualized antithrombotic therapy should be carried out according to particular case.

Objective The inhibitory effects of avastin on new blood vessels in nonproliferation diabetic retinopathy, age-related macular degeneration and neovascular glaucoma have been demonstrated. But only seldom report of avastin on corneal neovascularization(CNV) was seen. Present study was to evaluate the effect of topical bevacizumab (avastin) on experimental corneal neovascularization in mice. Methods Thirty eyes of 30 Balb/c mice were chemically cauterized by applying a 2 mm-diameter filter paper soaked 1 mol/L NaOH solution at the central cornea for 40 s. All animals were randomly assigned to five groups, including 1 mg/mL, 3 mg/mL and 5 mg/mL bevacizumab eye drops group respectively, 1 mg/mL dexamethasone sodium phosphate eye drops group (positive control) and normal saline solution group (negative control) . The drug was topically utilized twice per day. CNV was examined under the slim lamp on the 3rd, 7th and 14th day after alkali burn. Animals were killed on the 14th day after alkali burn. Area of CNV was calculated in terms of pixels on digital photographs. The use of animals followed the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission. Results No significant difference was found in the grade of corneal injury among five groups (F = 0. 201, P = 0. 935). The area of neovascularization at the cornea surface was (37.11 ±3.17)% in 1 mg/mL bevacizumab group, (29.75 ±3.56)% in 3 mg/mL bevacizumab group, (18. 76 ± 2. 55) % in 5 mg/mL bevacizumab group, (20. 91 ± 2. 75) % in dexamethasone group and (41. 65 ±2. 11)% in normal saline group, showing a significant difference among groups(F = 71. 687, P =0. 000) with the further comparative decline in 5 mg/mL bevacizumab group compared with other groups (P ＜ 0. 01) . Conclusion The topical use of bevacizumab (avastin) inhibits alkali burn-induced CNV in mice.

Objective To analyze the clinical results and early complications of corpectomy and reconstruction with titanium mesh cage implantation and pedicle screw fixation via a single posterior approach for severe thoracic and lumbar fractures.Methods Forty-four patients treated by reconstruction with titanium mesh cage implantation and pedicle screw fixation via a single posterior were studied retrospectively.There were 35 males and 9 females,with an average age of 37.3 years(range,19-66 years).The injury segments include 1 case at T11,5 cases at T12,20 cases at L1,11 cases at L2,5 cases at L3 and 2 cases at L4.According to AO classification,there were 24 cases of A3,17 cases of B1 and B2,and 3 cases of C1.According to ASIA,there were 10 cases of grade A,17 cases of grade B,10 cases of grade C and 7 cases of grade D.The neurologic function and effectiveness of correction of preoperative,immediate postoperative and 2years follow-up were compared,and the clinical outcome and early complications were analyzed.Results The follow-up time was 24 to 58 months,mean 38.9 months.At the time of 2 years postoperation,43 cases of incomplete neurologic deficit had improved 1 or 2 ASIA grades except 1 case of grade A.The results of decompression and reduction were satisfactory from the postoperative radiographic examinations.The correction maintained well and the implant loosening was not seen in 43 cases(97.7%)at the last follow-up.The com plications include:excessive blood loss(＞1500 ml)in 9 cases,transient nerve root injury in 4,cerebrospinal fluid leakage in 3,instrumentation failure in 1,mesh cage malposition in 3,iatrogenic leaving of free bone granula into the canal in 2,and superficial infection in 1.Conclusion This technique is effective for decompression and fusion,less invasive than combined anteroposterior procedure,and may be another good alternative for the treatment of severe thoracic and lumbar fractures.The early complications are not rare,but most of them are not serious and are relative to techniques.

Objective To explore the relationship between bone mineral density(BMD) and body tat disrribution in the aged in Ningbo area of China. Methods The BMD of lumbar vertebra and total body bones as well as body composition were measured by dual-energy X-ray absorptiometry (DXA) in 61 aged males, and also in 63 aged females as control group. Results In the aged males, a negative correlation was found between BMD and age, while BMI, muscle, trunk fat were positively correlated with BMD. As for the aged female, age, BMI, muscle and fat were all positively correlated with BMD. Conclusions The correlation between body fat and BMD is closer in females than in males in the aged. There is a positive correlation between trunk fat and lumbar spine BMD, and it indicates that central obesity may have a protective effect on lumbar spine BMD.

Objective To investigate the effects of blocking gastrin receptor on the proliferation,apoptosis and expression of key proteins in the related pathway in gastric cancer cell lines.Methods In the experimental group,the gastric cancer cell lines SGC-7901 and AGS cells were treated with 5 mmol/L proglumide,a kind of a gastrin receptor antagonist.And the normal cultured gastric cancer cells SGC-7901 and AGS were used in control group.The growth of each group was detected by MTT assay;the cell growth curve was drawn by flow cytometry;the cell cycle of each group was detected by flow cytometry.Annexin V-FITC/PI double staining was used to detect the cell growth of apoptosis.The relative mRNA expression of β-catenin,nuclear factor-P65,mammalian target of rapamycin and glycogen synthase kinase 3 beta in Wnt,NF-κB and PI3K-AKT-MTOR pathways were detected by RT-qPCR.The expression of β-catenin protein was detected by Western blotting.Results After treatment with proglumide,the growth of the cells in the experimental group was lower than that in the control group;and the proportion of S phase cells in the cell cycle was also lower than that in the control group,but the proportion of cells in G0/G1 phase was higher than that in the control group(P<0.05).The percentage of apoptotic cells was also increased after treatment with proglumide(P<0.05).Furthermore,proglumide treatment significantly reduced the expression of β-catenin at both mRNA and protein levels(P<0.05).Conclusion Blocking gastrin receptor can down-regulate the expression of β-catenin,inhibit the cell proliferation and promote the cell apoptosis in gastric cancer cells.

Objective To observe the influence of inactive FRMD4A gene′s expression on the biological behavior of tongue cancer CAL-27cell. Methods FRMD4A-siRNA was transfered into CAL-27 cell by lipidosome, to the expression of FRMD4A-siRNA in CAL-27 cell after transfection was detect by qRT-PCR cell proliferation , was checked by CCK-8,the influence of inactive FRMD4A gene on cell cycle distribution of CAL-27 cell was assayed by flow cytometry. Results Compared with the control group, FRMD4A mRNA expression significantly reduced in FRMD4A-siRNA interfering group (94%) and the cell proliferation index decreased(P<0.05). The cell cycle arrested in G1 period (P<0.05). Conclusion FRMD4A-siRNA could effectively inhibit FRMD4A mRNA expression in tongue cancer CAL-27cell, impact the distribution of cell cycle, and reduce cell proliferation.