AIM: To evaluate the effects of all-trans retinoic acid (atRA) on the proliferation in cultured mouse cerebral microvascular endothelial cells (bEnd.3) . METHODS: Cultured cells were divided into five groups randomly, one as control group, the other four groups were 10-9, 10-8, 10-7 and 10-6 mol/L group. Effects of atRA on proliferation in bEnd.3 cells were detected by flow cytometry and immunocytochemitry of PCNA and MTT at 24 h, 48 h and 72 h. The effects of atRA (10-6 mol/L group) on the expressions of angiogenic genes in bEnd.3 cells were studied using microarray. RESULTS: The results of MTT and flow cytometry showed that all- trans retinoic acid at concentration of 10-6 mol/L significantly inhibited the proliferation of bEnd.3 cells. Immunocytochemical staining showed the expression of PCNA was markedly decreased in bEnd.3 cells at 24 h after treatment with atRA. Microarray results demonstrated that there were 11 down - regulated angiogenic genes and 2 up - regulated angiogenic genes in 10-6mol/L atRA group. CONCLUSION: All - trans retinoic acid at concentration of 10-6mol/L may significantly inhibit the proliferation of bEnd.3 cells treated for 24 h ire vitro via down-regulation of angiogenic genes and PCNA expression.

Objective:To do research on prediction method of medical equipment sudden fault for the requirement of medical security at sea which is away from base.Methods: Sudden fault data of the medical equipment is a random variable which often manifest as the time of sudden fault happening. The possible distribution type of fault data is hypothesized according to the engineering experience. In order to ensure the sudden fault density function of medical equipment, parameter estimation and distribution fit test are carried out for the fault data.Results: The sudden fault prediction model of medical equipment is established to get the future sudden fault probability of medical equipment based on the distribution function of fault data.Conclusion: The results of case analysis validate the rationality of sudden fault prediction model.

Objective To explore the value of echocardiography in the diagnosis of anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) in children.Methods The echocardiographic images of 14 patients with ALCAPA confirmed by operation and 18 patients with endocardial fibroelastosis (EFE) diagnosed by clinical were compared and analyzed.Results Both ALCAPA and EFE exhibited obvious dilated left ventricle,decreased left ventricular systolic function,thick endocardium and mitral regurgitation of different degree.The former additionally showed dilated right coronary artery(RCA) with normal origin,left coronary artery(LCA) emerging from the root or wall of the pulmonary artery(PA),the retrograde flow into PA in LCA and abundant collateral vessels in myocardium.However the later exhibited normal diameter of LCA and RCA and no collateral vessels.Conclusions Color Doppler echocardiography not only demonstrates left ventricular systolic function,endocardium,mitrial regurgitation and collateral vessels in myocardium,but also shows the origination and courses of LCA clearly,which provide exact informations to diagnose ALCAPA.The echocardiography can be used as a powerful tool of ALCAPA diagnosis and preoperative assessment.

Microvascular endothelial cell (MVEC) is one of the target cells of TNFα (TNF effect). The dysfunction of MVEC induced by TNF plays an important role in some cardio-cerebral vascular diseases. ① Cell proliferation kinetic: Using flow cytometry, we found cell count ［(4.30±0.34)×107／L)］ in TNF group (4×105 U/L) was obviously less than that in control ［(5.23±0.50)×107／L, P＜0.01］. The cells of G1 phase were more than those of the control, while the cells of G2, S and M phase became less (P＜0.05). ② Coagulant and anticoagulant: 72 h after MVEC cultued in the media, the content of 6-keto-PGF1α (RIA) and activity of PAI decreased significantly in TNF (4×105 U/L) group (P＜0.01, vs control). The difference between TXB2 content and t-PA activity in groups was not significant (P＞0.05). ③ Adhesive molecule: The effect of low concentration TNF (＜4×105 U/L) on adhesion between cultured MVEC and leukocytes was not signficant, but when the concentration of TNF reached 8×105 U/L or more, 12 h after culture the adhesion rate between MVEC and neutrophil increased 30.8%±4.5%. If adding monoclonal antibody of ICAM-1/CD11 into media, the adhesion rate of leukocytes decreased significantly (from 31.2% to 63.4%). ④ NO: The level of nitrite in culture media (Griess reaction) was higher than that of control (P＜0.05) after pretreatment of TNF (2×106 U/L) for 6 h. Adding L-NMMA, Dexamethasone or Cycloheximide in media could block the increase of nitrite induced by TNF, while L-Arg could enhance it. The expression of iNOS mRNA of PMVEC increased significantly after treated with TNF (2×106 U/L) for 24 h (quantitative RT/PCR). Pretreatment with Dexamethasone or Cycloheximide could block the increase (P＜0.05). Meanwhile, the expression of eNOS mRNA decreased significantly compared with control, the decrease can be blocked by Cycloheximide but not by Dexamethasone. So that TNF can induce the expression of iNOS mRNA in PMVEC, but inhibited the expression of eNOS mRNA. NO production in PMVEC can be time-dependently induced by TNF. ⑤ Apoptosis: Adding high concentration TNF(＞1.2×106 U/L) in culture media for 12 h can induce apoptosis of PMVEC with electron microscopy, flow cytometry (PI/AnnexinV stain), TUNEL and DNA ladder eletrophoresis. Meanwhile, expression of apoptosis-associated gene Bcl-2 mRNA decreased and that of Fas mRNA increased (Northern blot). The expression of FADD, caspase 3 and caspase 8 enhanced too. So that signal of apoptosis induced by TNF may be transmitted following the cascade of Fas→FADD→caspase8→caspase3. In conclusion, it should be paid attention to metabolism inability and dysfunction induced by TNF which can not found easyily using light microscope. High concentration TNF can induce apoptosis of endothelial cells regulated by apoptosis-associated genes. The changes mentioned above are common pathway in pathogenesis of some diseases related to TNF.

Objective To study the echocardiographic characteristics and its diagnosis value on congenital supravalvular aortic stenosis (SVAS) in children. Methods Thirty-one patients with SVAS diagnosed by multiplane echocardiography were enrolled in the study. Their echocardiographic characteristics were compared with cardiac catheterization, operation, and gene detection results. Echocardiographic changes were mainly observed in aortic valve, supravalve, descending aortic arch, pulmonary artery valve, main pulmonary artery and its branches,and coronary artery. Results Of the 31 patients,26 had hourglass type SVAS,4 hypoplastic type,and 1 membranous type; 2 patients had extremely mild stenosis (defined as a Doppler gradient <25 mm Hg) ,20 mild (25～49 mm Hg) ,5 moderate (50～75 mm Hg) ,and 4 severe C>75 mm Hg) ones. Nineteen patients were diagnosed with Williams syndrome by gene detection. Three patients were associated with aortic valve stenosis including one missed at the initial diagnosis; 10(32.26%) patients with pulmonary stenosis, including pulmonary valve stenosis in 6, left and right pulmonary artery stenosis in 3 ,and branch stenosis in 1:6 patients with coronary stenosis. Conclusions The sternal border and five chamber apical views are the best to detect SVAS. Williams syndrome patients are prone to SVAS.Pulmonary stenosis echocardiography forms a great proportion of the SVAS patients. Routine examination is necessary for coronary stenosis in cases of SVAS.

AIM: To establish a quantitative measurement method of ECV-304 cell migration in a scratch wound model in vitro. METHODS: The method of ECV-304 cell scratch wound model was improved. The ECV-304 cell migration was measured quantitatively using a computer-assisted video microscopic method with image-analysis system. The effects of heparin and thrombospondin-1 on ECV-304 cell migration were observed in the scratch wound model in vitro. RESULTS: The average width and length of cell scratch wound was (0.95?0.08) mm and (51.20?3.40) mm respectively in 120 experiments. The peak of cell migration reached about 24 h after culture. The stimulated effect of heparin and inhibited effect of thrombospondin-1(TSP-1) on cell migration were observed. CONCLUSION: A quantitative measurement method of ECV-304 cell migration in vitro is established. The stimulated and inhibited effects on ECV-304 cell migration were proven respectively using this method.

AIM: To evaluate the changes of serum total prostate specific antigen (T-PSA), free prostate specific antigen (F-PSA) and the ratio of F-PSA to T-PSA (F/T) in patients with prostate cancer (PCa) and its clinical significance. METHODS: The concentrations of T-PSA and F-PSA in serum were measured by micropartical enzyme immunoassay (MEIA) using AxSYM System, and the F/T ratio was calculated. RESULTS: Before operation, the concentrations of T-PSA and F-PSA in patients with PCa were much higher and F/T ratio was significantly lower than that in patients with benign prostate hyperplasia (BPH). T-PSA and F-PSA levels decreased, but F/T ratio increased after operation in PCa and BPH. F/T ratio in 83.5% PCa and 6.5% BPH was less than 0.16. To diagnosis PCa, the sensitivity of F/T ratio was 83.5%, and the specificity was 86.7%. CONCLUSION: Serum T-PSA, F-PSA and F/T ratio are important parameters for the early diagnosis of prostate cancer. [

AIM: To study the cytological characteristics and gene expression of normal cultured bEnd.3, a mouse brain microvascular endothelial cell strain. METHODS: The morphology of bEnd.3 was studied by light and electronic microscopy, its molecular markers were observed by immunocytochemistry. Cell proliferation kinetics and apoptosis were analyzed by flow cytometry and MTT assay, PGE_2 level was measured by ELISA, and expression of the genes that closely related with vascular endothelial functions was studied by gene micro-array. RESULTS: bEnd.3 had morphological characteristics of microvascular endothelial cells (MVEC) growing in a cobblestone pattern, forming tube-like structure or capillary network and having microvilli. Furthermore, bEnd.3 showed positive staining for vW factor and CD34 and secreted high level of PGE_2 (644.55?30.24 ng/L). Gene micro-array analysis showed CD31, CD36, CD105 expression, and other genes closely related to microvascular endothelial functions also expressed at relatively high level. In addition, bEnd.3 responsed sensitively to mitogen such as basic fibroblast growth factor. CONCLUSION: bEnd.3 is a kind of MVEC, and it can be utilized to study the mechanisms of some diseases such as cancers and cardio- cerebral vascular diseases.

Objective To report the clinical and imaging characteristics of 9 cases of gastric duplication in children,as well as to probe into all kinds of imaging examination methods,with an aim to improve preoperative diagnostic accuracy.Methods Nine cases of children with gastric duplication that received surgical resection and were pathologically confirmed in our hospital from January 2009 to November 2016 were retrospectively analyzed.The clinical manifestations,complications,complicated malformation and imaging findings of the cases were summarized.Three asymptomatic cases were carried out regular ultrasound examination follows-up preoperatively.Results Gastric duplication in children varied in terms of cliuical manifestations.All the 9 cases were performed ultrasonography preoperatively,and 5 of them were accurately diagnosed.Meanwhile,7 cases received preoperative CT examination,with 2 cases being confirmed.Three cases received magnetic resonance imaging (MRI) examination,with 2 cases being confirmed.And 2 cases had upper abdominal gastrointestinal imaging (GI) examination,but no definite diagnosis was made.Three cases of asymptomatic children with enlarged lesion can be seen in follow-up.Conclusions Gastric duplication is a rare congenital malformation,ultrasound allows to clearly display the capsule wall structure and the relation between gastric duplication with gastric wall,which is associated with high utilization rate and diagnosis rate,and it can be served as the preferred imaging examination method for gastric duplication in children.