A simple and effective method for the duplex PCR detection was developed by using sequences of exogenous gene(RDV MP-)and endogenous gene(Zein)as templates for PCR amplification.The results of routine PCR amplification for RDV MP-gene in transgenic maize suggested that RDV MP-gene can stably inheritate in transgenic plants and their progenies;The duplex PCR detection of all negative and part positive samples that obtained by routine PCR amplification confirmed that above negative results were exact,also showed that the quality of extracted DNA can meet the need of PCR amplification.The error ratio of negative samples was 1.4%.The method used in this study was simple and credible and can be used to detect transgenic plants and their products.

Background and purpose:The lymph node metastatic model of rectal tumor is a useful tool for the research on tumor occurrence, development, metastasis and antineoplastic therapy. There are few reports about establishment of larger animal model. This study aimed to establish feasible and reproducible lymph node metastatic models of VX2 tumor in rabbits.Methods:The VX2 tumor tissue was put into the puncture needle. The VX2 tumor tissue in the needle was orthotopically transplanted into the rectal wall of the New Zealand white rabbits successfully. Twenty New Zealand white rabbits were transplanted. Two experimental rabbits were scanned by MR weekly. Tumor growth curve and lymph node numbers were observed on MR. Experimental rabbit tumor volumes were measured by MR post-processing software. The rectal tumor and surrounding lymph nodes were resected, and the specimens were ifxed. The sections were stained with HE. We explored the relationship between tumor volume and growth time, the number of metastatic lymph nodes and tumor volume, respectively.Results:Thirteen models were successfully established with a rate of 65%. Tumors limited in the rectal wall were observed on the fourth week. Tumor size increased over time. There was significant difference in the tumor volume between different periods (growth cycle number) (F=52.865,P<0.05). There was a signiifcantly positive correlation between tumor volume and the growth cycle number (r=0.910,P<0.05). The metastatic lymph nodes could be observed when VT>9 cm3. The number of metastatic lymph node increased obviously from the ninth week. The more tumor volume, the greater the number of metastatic lymph nodes was observed (F=92.531,P<0.05). There was a signiifcantly positive correlation between the number of metastatic lymph nodes and the tumor volume (r=0.945,P<0.05).Conclusion:Metastatic lymph node models of VX2 tumor in New Zealand white rabbits were established successfully. This model has some value in the research on local growth, invasion mechanism, lymph node metastasis and biological characteristics of rectal cancer.

AIM: To evaluate the therapeutic efficacy of intratracheal instillation of porcine pulmonary surfactant(PPS) in rats with lipopolysaccharides(LPS)-induced early-stage ALI in this study.METHODS: SD rats weighing 200 g-300 g were randomly divided into 4 groups: LPS(1.5 mg?kg-1)+saline,LPS+PPS 100 mg?kg-1,LPS+PPS 150 mg?kg-1,LPS+PPS 200 mg?kg-1.The PaO2 and PaCO2,as well as survival rate of rats were examined for 6 h after the start of PPS-instillation.Then,rats were killed and lungs were immediately removed for lung index(LI) and histological analysis.The bronchoalveolar lavage fluid(BALF) was collected for measurement of total protein(TP) contents,TNF-? level and white blood cell(WBC) numbers.RESULTS: Significantly increased PaO2,reduced mortality rate,decreased total protein and TNF-? contents in BAL,as well as lung index and meliorated histological appearance were observed in three PPS-treated groups compared with group given saline after LPS(P

Objective To compare two methods 3D flash and diffusion-weighted images(DWI)in reconstructing the brain surface anatomy,and to evaluate their displaying ability,advantages,limitations and clinical application.Methods Thrity normal cases were prospectively examined with 3 D flash sequence and echo-planar DWI.Three-dimensional images were acquired with volume-rendering on workstation.Brain surface structures were evaluated and scored by a group of doctors.Results Main structures of brain surface were clearly displayed on three-dimensional images based on 3D flash sequence.Average scores were all above 2.50.For images based on DWI,precentral gyrus,postcentral gyrus,superior parietal lobule, superior frontal gyrus,precentral sulcus,central sulcus,postcentral sulcus,intraparietal sulcus and superior frontal sulcus were best shown with average scores between 2.60-2.75,However,supramarginal gyrus, angular gyurs,middle frontal gyrus,inferior frontal gyrus,superior temporal gyrus,lateral sulcus,inferior frontal sulcus could not be well shown,with average scores between 1.67-2.48.Middle temporal gyrus, inferior temporal gyrus,superior temporal sulcus and inferior temporal sulcus can only get scores from 0.88 to 1.27.Scores of images based on 3D flash were much higher than that based on DWI with distinct differentiations,P values were all below 0.01.Conclusion Three-dimensional images based on 3D flash can really display brain surface structures.It is very useful for anatomic researches.Three-dimensional reconstruction of brain surface based on DWI is a worthy technique to display brain surface anatomy, especially for frontal and parietal structures.

Objective To compare the different expressions of transforming growth factor beta1 (TGFβ1) and its type one receptor(TGFβR1) between primary pancreatic cancer cell line AsPC-1 and metastatic pancreatic cancer cell line BxPC-3. Methods The mRNA expressions of TGFβ1 and TGFβR1 in AsPC-1 and BxPC-3 pancreatic carcer cell lines were quantatitived by real-time RT-PCR. The protein levels of TGFβ1 and TGFβR1 in these two cell lines were measured by Western blot assay. Results Compared with the primary pancreatic cancer cell AsPC-1, the mRNA and protein expressions of TGFβ1 and TGFβR1 were much higher in BxPC-3 pancreatic cancer cells(P<0.05). Conclusions Upregulations of TGFβ1 and TGFβR1 might be a pivotal incidence in the procedure of malignant progressing and metastasis in pancreatic cancer cells.

High blood pressure (hypertension) is implicated in the development of atherosclerosis. Blood vessels are constantly subjected to stretch due to blood pressure and changes in stretch usually instigate adaptive vascular remodeling, including abnormal growth and proliferation of vascular smooth muscle cells (VSMCs) as well as extracellular matrix (ECM). In this experiment, we used bovine aortic endothelial cells and smooth muscle cells (EC-SMC) co-cultured ePTFE vascular grafts subjected to normal atmospheric pressure (as a control), and 100 mmHg hydrostatic pressure for 7 d. The increase of cell layer thickness was observed. When measured, the cell layer thickness increased by 116.2%. The increase of collagen (Type IV) synthesis was also observed in the immunohistochemistry assay. When stained with toluidine blue, the cells showed metachromatic phenomenon.
Animals ; Blood Vessel Prosthesis ; Cattle ; Cell Culture Techniques ; methods ; Cell Proliferation ; Cells, Cultured ; Collagen Type IV ; metabolism ; Hydrostatic Pressure ; Mechanotransduction, Cellular ; physiology ; Muscle, Smooth, Vascular ; cytology ; physiology ; Polytetrafluoroethylene ; chemistry ; Tissue Engineering ; methods ; Transplants

ObjectiveTo investigate the effects of infrared radiation and magnetic fields on the synovitis of rabbit with experimental knee osteoarthritis (OA).MethodsTwenty-four New Zealand grown rabbits were duplicated for OA model of the knee by extended fixation technique, and after 6 weeks, all rabbits were unchained from fixation. The rabbits were randomly divided into four groups: control group (no treatment), infrared group (treated with infrared radiation ), magnet-therapy group (treated with magnetic fields ) and the combined therapy group (treated with infrared radiation and magnetic fields). At the ends of the first, second and third week after treatment, 2 rabbits in each group were taken to measure the general change and histopathology of their synovium.ResultsCompared with other groups, the severity of synovitis (include the synovium line cells proliferation, the invading of granulation and blood vessels in the underlayer of synovium, the infiltration of inflammatory cells) declined significantly in combined therapy group. Scores of Ayral's index of combined group were significantly lower than other groups ( P<0.05), and there was no significant difference between infrared group and magnet-therapy group ( P>0.05).ConclusionThe combination of infrared radiation and magnetic fields can efficiently suppress the synovitis of OA and alleviate the tissue's destructions.

Objective To understand the information of food hypersensitivity of children with Henoch-Schonlein purpura (HSP) by food intolerance test.Methods Seventy-four children (40 boys and 34 girls;aged from 3 to 14) with HSP who were hospitalized in our hospital from Dec.2006 to May 2007 were chosen.Two mils venous blood was collected from each object,separated serum.Enzymelinked immunosorbent assay method was applied to detect the serum concentration of 14 kinds food allergen IgG in 74 children with HSP.The concentration of food allergen IgG was divided into 4 levels: 0(IgG0.20 U/L,servious allergy).Results The positive rate was 77.0% in 74 cases with HSP.One type of food allergen specific IgG increased in 24 cases(42.1%),2 types of antibodies increased in 16 cases(28.1%),3 types of antibodies increased in 12 cases(21.1%).Four or more types of antibodies increased in 5 cases(8.7%).The positive rates of 14 food allergen IgG were as follow:egg 93.0%,milk 26.3%,soybean 15.8%,tomato 14.0%,wheat 12.3%,ling 12.3%,shrimp 8.8%,crab 8.8%,beef 3.5%,corn 1.8%,rice 1.8%.Conclusion Food intolerance test is a useful method for children with HSP to find food allergen and guideline for diet of these children.

Child ; Humans ; Male ; Muscles ; pathology ; physiopathology ; Myopathies, Structural, Congenital ; diagnosis ; Prognosis

ObjectiveTo explore the clinical features and factors inlfuencing the prognosis of childhood non-Hodgkin's lymphoma (NHL).MethodsPathologically diagnosed 78 pediatric patients with NHL and treated in the Afifliated Hospital of Qingdao University from January 2004 to August 2013 were collected and analyzed. Patients were grouped according to age, sex, tumor size, immunologic classiifcation, B-symptoms, LDH, hemoglobin and clinical staging. The 5-years event-free survival rate (EFS) were calculated and analyzed by Kaplan-Meier method, and the difference of the survival rate between groups were com-pared. Using Cox proportional hazards model, we analyzed the possible factors that might inlfuence 5-years event-free survival rate EFS , such as age and clinical staging. TheOR value and the 95%CI were calculated.ResultsAmong the 78 cases, median age of onset is 7 years old, male to female ratio is 2.90:1, there are 25 cases of T-cell type and 53 cases of B-cell type. According to pathological types,Burkitt lymphoma is the most common (34.6%), followed by T-lymphoblastic lymphoma (20.5%), diffuse large B-cell lymphoma (11.5%). According to the St. Jude malignant lymphoma staging system, there are 2 cases in stage I, 9 in stageⅡ, 35 in stageⅢ and 32 in stageⅣ. Swelling of periphery lymph node (80.7%) was observed as initial symptom in 26 cases of lymphoblastic lymphoma. Among 45 cases of mature B-cell tumor, the main clinical feature including abdominal cavity and gingival were observed in 27 cases of Burkitt lymphoma. Among the 73 cases received treatments, 66 cases (90.5%) attained CR (complete remission) and 4 cases (5.5%) attained PR (partial remission) by cytology and radiographic assessment after two course of combined chemotherapy, 2 cases (2.7%) rapidly relapsed after the remisson of one course treatment, 1 case (1.3%) appeared the central nervous system inifltration in the chemotherapy. With median follow-up time of 42 months, the 5-year EFS of the 73 cases was (67.0+5.5)%. Single factor analysis showed that B-symptom, LDH, and clinical staging were signiifcantly correlated with prognosis (P<0.05), while age, sex, tumor size, hemoglobin and immune classiifcation was independent of prog-nosis (P>0.05). Multiple factor analysis showed that LDH and clinical staging inlfuenced the prognosis (OR=3.34,95%CI 2.275?10.683,P<0.01;OR=4.354,95%CI 1.519?12.475,P<0.01) .Conclusionclinical features of childhood NHL are variable. LDH and clinical staging at primary diagnosis are important factors affecting the prognosis.