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.

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

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.

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 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.

BACKGROUND:In vitro experiments have demonstrated that the biodegradable mesh-like microporous baloon made of macromolecular materials has obvious advantage of anti-leakage, which is capable of maintaining calcium homeostasis, has no inhibitory effects on cel growth and on microscopic interdigitation formation between new bone and bone cement. OBJECTIVE:To evaluate the therapeutic effects of biodegradable mesh-like microporous baloon with calcium bone cement on vertebral fractures based on animal experiments. METHODS:The fracture model was established in 48 New Zealand rabbits, in which a bone dril was introduced after successful puncture at sites near left low extremity of the femur. These rabbit models were randomized into two groups: experimental group with calcium phosphate bone cement and biodegradable mesh-like microporous baloon and control group only with calcium phosphate bone cement. Clinical parameters such as blood cel count, biochemistry, and CT/X ray were examined at 1, 3 and 6 months after implantation of the baloon and bone cement. After that, the specimens were fixed for pathological analysis. RESULTS AND CONCLUSION:The operation was performed under general anesthesia with no eventful infusion of bone cement. The expansion of baloon was satisfactory without definite extravasation of bone cement in the experimental group. In the control group, cement diffusion was found with pulmonary embolism occurring in three New Zealand rabbits. No statistical significance for blood cel counts and biochemistry was found between pre- and postoperation or between two groups. The materials in the two groups had favorable biocompatibility with injured bones without obvious immunological response. In the experimental group, the baloon wal was thinned and partial bone tissues grew into the cement at 1 month; at 3 months, a large amount of bone tissues grew into the cement and cement volume diminished; at 6 months, the baloon disappeared and only a smal amount of cement left in the bone tissues. In the control group, it was difficult to determine when the cement degraded. The biodegradable mesh-like microporous baloon combined with calcium bone cement is superior to bone cement alone in the management of vertebral fractures.

Objective To explore the immune effects of high intensity focused ultrasound (HIFU) in the treatment of pancreatic carcinoma and to investigate the imaging methods to evaluate HIFU's efficacy.Methods A total of 32 patients with pancreatic carcinoma treated by HIFU were enrolled.The freeze-thaw antigen was prepared by freezing and thawing the cancer cells.HIFU antigen was prepared by cancer cells sonicated by HIFU.The killing effects of no antigen activated dendritic cells (DC) induced T lymphocyte (DC-T),freeze-thaw antigen activated DC induced T lymphocyte (freeze-thaw antigen-DC-T) and HIFU activated DC induced T lymphocyte (HIFUantigen-DC-T) in autologous pancreatic cancer cells were detected by lactic dehydrogenase kit.The changes of immune indexes [heat shock protein 70 (HSP70),T helper lymphocyte Thl/Th2 and transforming growth factor-β (TGF-β)] before and after H IFU treatment were determined by enzymelinked immunosorbnent assay (ELISA) method.The changes of clinical efficacy indexes [visual analogue scale (VAS),performance status (PS) and carbohydrate antigen (CA) 19-9] before and after HIFU treatment were compared.The instant and recent (two months) efficacy of HIFU treatment were evaluated by contrast enhanced ultrasonograph (CEUS) and computed tomography (CT).The line q test was performed for comparision between groups.t-test was applied for comparision before and after treatment.Results Compared with freeze-thaw antigen,the killing effect of HIFU antigen-DC-T in autologous pancreatic cancer cells was higher (40.24％ ± 10.56％ vs 46.93％±13.26％,q=3.44,P＜0.05).HSP70 [(17.31±4.75) ng/mlvs (22.84±5.56) ng/ml],Th1/Th2 (1.24±0.36 vs 1.47±0.31),TGF-β [(1.39±0.41) ng/ml vs (1.04±0.38) ng/ml],VAS (3.97±1.32 vs 3.26±1.18),PS (2.76± 1.02 vs 2.21±0.86) and CA19-9 level[(135.39±37.45) U/ml vs (114.82±30.51) U/ml] improved after HIFU treatment compared with those before treatment (t=4.278,2.739,3.542,2.268,2.332 and 2.409,allP＜0.05).CEUS and CT showed that blood supply and the volume of the tumors reduced after HIFU treatment.Conclusions HIFU is effective in treating pancreatic carcinoma,improving immune status of patients and enhancing antitumor response.CEUS can real-time evaluate the efficacy of HIFU treatment.

BACKGROUND:Vertebroplasty and kyphoplasty for osteoporotic vertebral compression fractures can result in many complications, such as bone cement leakage and adjacent-level fractures.
OBJECTIVE:To verify the possibility of biodegradable mesh-like microporous polymer bal oon for the treatment of osteoporotic vertebral compression fractures.
METHODS:Biodegradable mesh-like microporous P(DLLA-CL) bal oons were fabricated by electrospinning technique. Coated bal oons with the same specification was fabricated by coating P(DLLA-CL) onto the same mould. Morphology of the bal oons was observed by scanning electron microscopy. The bal oon leakage was observed by eyes after the injection of water or cement. The initial strength and stiffness were measured by a universal testing machine. The proliferation of MC3T3-E1 cel s on the bal oons was determined by laser confocal microscope and cel counting kit-8 assays. The biodegradation of bal oons in simulated body fluid, porcine pancreatic lipase, and fresh human serum was studied by residual weighing and scanning electron microscopy observation. Burst pressure of bal oons was measured after the bal oon was placed into a hole in the vertebral bone. For the in vitro calcium release tests, the bal oons were fil ed with calcium cement, tied, placed into 6atm ultrapure water, and then the calcium concentration was regularly determined.
RESULTS AND CONCLUSION:Mesh-like microporous bal oons presented with good fiber morphology, thickness distribution, and the presence of pores;on the coated bal oon surface, there was absence of specific morphology and porosity. Compared with the coated bal oon, the mesh-like microporous bal oon showed better mechanical properties, liquid permeability and burst pressure, to prevent leakage of bone cement and promote osteoblast adhesion and proliferation. In addition, the degradation of the mesh-like microporous bal oons was more uniform and stable than the coated bal oons, which may increase the calcium concentration in the injured vertebrae and wil be beneficial to the new bone growth and fracture healing.