Objective To investigate the relationship between T?cell death?associated gene 8 ( TD?AG8) and endogenous neuron?protective mechanism against oxygen?glucose deprivation and restoration ( OGD∕R)?induced apoptosis in rat neurons. Methods The primary cortical neurons obtained from fetal rats were seeded in 6?well plates at a density of 1×105 cells∕ml and divided into 5 groups using a random number table: control group ( group C, n=24 ) , group OGD∕R ( n=48 ) , TDAG8 agonist BTB09089 group (group BTB, n=24), TDAG8?siRNA group ( group siRNA, n=24), and blank vehicle group ( group V, n=24) . The medium was replaced with glucose?and serum?free Locke′s buffer, and the neu?rons were exposed to 95% N2?5% CO2 in an air?tight incubator at 37℃ for 60 min followed by routine cul?ture to establish the model of OGD∕R. In BTB, siRNA and V groups, 20 μmol∕L TDAG8 agonist
BTB09089, 200 pmol∕L TDAG8?siRNA, and 6 μl∕200 μl transfection reagent were added, respectively, at 24 h before oxygen?glucose restoration. At 6 h of oxygen?glucose restoration, the neuronal viability and a?mount of lactic dehydrogenase ( LDH) released were measured, and the expression of TDAG8 and caspase?3 mRNA in neurons was detected by fluorescent quantitative real?time polymerase chain reaction. In group OGD∕R, the expression of TDAG8 and caspase?3 was measured by Western blot at 0, 3, 6, 12 and 24 h of oxygen?glucose restoration. In C, OGD∕R, BTB, siRNA and V groups, the expression of TDAG8, caspase?3 and p?Akt was detected at 6 h of oxygen?glucose restoration. Results In group OGD∕R, the ex?pression of TDAG8 was gradually up?regulated after oxygen?glucose restoration, and the expression of caspase?3 peaked at 6 h of oxygen?glucose restoration. Compared with group C, the neuronal viability was significantly decreased, the amount of LDH released was significantly increased, and the expression of TD?AG8 and caspase?3 protein and mRNA and p?Akt was significantly up?regulated in OGD∕R, V and siRNA groups ( P<0?05) . Compared with group OGD∕R, the expression of TDAG8 protein and mRNA and p?Akt was significantly up?regulated, the expression of caspase?3 protein and mRNA was significantly down?regu?lated, the neuronal viability was significantly increased, and the amount of LDH released was significantly decreased in group BTB, the expression of TDAG8 protein and mRNA and p?Akt was significantly down?regulated, the expression of caspase?3 protein and mRNA was significantly up?regulated, the neuronal via?bility was significantly decreased, and the amount of LDH released was significantly increased in group siR?NA ( P<0?05) , and no significant change was found in the parameters mentioned above in group V ( P>0?05) . Conclusion TDAG8 is partially involved in the endogenous neuron?protective mechanism against OGD∕R?induced apoptosis in rat neurons, which may be related to activation of Akt signaling pathway.

Objective To investigate the value of two-dimensional and three-dimensional transesophageal echocardiography (3DTEE) in transcatheter aortic valve implantation (TAVI). Methods Eleven patients with severe aortic stenosis and one patient with moderate to severe prosthetic valve regurgitation underwent TAVI in Zhongshan Hospital of Fudan University from May 2010 to December 2015. All patients received two-dimensional and three-dimensional transthoracic and transesophageal echocardiographic examination before surgery, during and after surgery.Results Procedural success was achieved in eleven patients, but one patient died in pericardial tamponade and aortic dissection three days after TAVI. The maximum and minimum diameter of aortic annule, the area of aortic annule and aortic valve were measured by multidetector computed tomography (MDCT) and 3DTEE before surgery. All parameters had strong correlations between MDCT and 3DTEE (r=0.98,P<0.01 for maximum diameter;r=0.97,P<0.01 for minimum diameter;r=0.97,P<0.01 for the area of aortic annule;r=0.99, P<0.01 for the area of aortic valve). There were good correlations for the area of aortic valve among MDCT, 3DTEE and equation of continuity (allr=0.99,P<0.01).ConclusionTwo-dimensional and three dimensional transesophageal echocardiography can quantify the size of aortic annular and comprehensively evaluate the anatomical structure of aorta rapidly and accurately, which can be used in guiding TAVI and monitoring its complications in real time.

In the criminal cases of driving under the influence (DUI), DNA evidence can be collected from the deployed airbag of the motor vehicle and submitted to the crime lab for touch DNA analysis. The evidence can be acquired when the skin cells are observed on the surface of the airbag in a traffic accident. However, the low quantity or quality of the evidence collected from a crime scene prevents further identification analysis in many cases. In the current study, we reported a case of identifying touch DNA extraction from the shed skin cells from the deployed airbag of a motor vehicle. We managed to collect DNA evidence from the shed skin cells in an airbag using a proper approach of collection and extraction. The 5.87 ng of extracted DNA was sufficient for genotyping and forensic identification, which helped to identify the driver of the car in collision with a pier in the street. In DUI cases and other traffic accidents, therefore, the amount of touch DNA extracted from the deployed airbag can be sufficient for DNA marker genotyping and further analysis.
Accidents, Traffic ; Air Bags ; Alcoholic Intoxication ; Crime ; DNA/analysis* ; Genotype ; Humans ; Motor Vehicles ; Skin/cytology* ; Touch

Objective To investigate the difference between three-dimensional transesophageal echocardiography ( 3DTEE) and CT in measuring the size of aortic ring and the height of coronary ostium . Methods Fifteen patients were recruited and were treated with the transcatheter aortic valve implantation ( TAVI) . Routine transthoracic echocardiography ,two-dimensional echocardiography ( 2DTEE) ,3DTEE and CT examinations were taken preoperatively . Results The minimal diameter ,maximum diameter , perimeter ,area of the aortic ring measured by 3DTTE showed a close correlation and a strong consistency with those measured by CT [ r = 0 .88 , P < 0 .0001 ,ICC = 0 .928 (0 .788 - 0 .976) ; r = 0 .81 , P = 0 .0003 , ICC = 0 .890 ( 0 .673 - 0 .963) , r = 0 .85 , P = 0 .0001 ,ICC = 0 .914 ( 0 .744 - 0 .971) ; r = 0 .88 , P <0 .0001 ,ICC = 0 .932 (0 .799 - 0 .977) ] .The ostium height of the left and right coronary arteries measured by 3DTEE also showed a close correlation and a strong consistency with those measured by MDCT [ r =0 .87 , P < 0 .0001 ,ICC = 0 .923 ( 0 .777 - 0 .975) ; r = 0 .82 , P < 0 .0002 ,ICC = 0 .897 ( 0 .693 - 0 .965) ] . Besides ,inter-observer and intra-observer reproducibility for 3DTEE measurement data were very good . Conclusions 3DTEE has a high repeatability in evaluating minimal diameter , maximum diameter , perimeter ,area of the aortic ring and the height of coronary ostium ,which also shows good correlation with those measured by CT .

OBJECTIVETo measure mobility of dendritic filopodia, complexity of dendritic arborization using method of live imaging in cultured rat hippocampal neurons and to analyze their morphological characters quantitatively.

METHODSVectors expressing Green Fluorescent Protein- Fibrous Actin (GFP-F-Actin) and F-GFP were co-transfected into cultured rat hippocampal neurons at 5 d in vitro (DIV 5). Neurons expressing GFP were photographed and analyzed with Metamorph software.

RESULTDendritic filopodia was observed to move actively from DIV 7 to DIV 9. The mean density of filopodia was (10.78 +/-3.78)/100 microm, (10.68 +/-2.96)/100 microm and (9.99 +/-3.67)/100 microm (P >0.05), and there were (30.18 +/-14.03)% to (87.36 +/-20.88)% filopodia were mobile (P <0.001). During DIV 7-DIV 14, the total length of dendritic branches grew from (410.74 +/-185.98) microm to (1238.21 +/-418.32)microm (P <0.001) and the number of dendritic branches increased from 18.93 +/-7.23 to 33.60 +/-10.46 (P<0.001). The density of spine was (37.17 +/-6.46)/100 microm at DIV 14.

CONCLUSIONThe combination of live imaging with quantitative analysis is a useful method to study dendritic morphological development in vitro, including indicators of dendritic filopodia, dendritic arborization and spines.

Actins ; genetics ; metabolism ; Animals ; Animals, Newborn ; Cells, Cultured ; Dendrites ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; Hippocampus ; cytology ; Microscopy, Fluorescence ; Neurons ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection

Objective To investigate the proportion and function of CD_4~+ CD_(25)~+ regulatory T cells (CD_4~+ CD_(25)~+ Tr)in unexplained recurrent spontaneous abortion(URSA).Methods(1)Proportion measurement:the proportion of CD_4~+ CD_(25)~+ Tr cells in peripheral blood was measured by double-label flow cytometric analysis.The samples were taken from 15 URSA women,15 normal non-pregnancy women and 13 normal pregnancy women.(2)Function measurement:CD_4~+ CD_(25)~+ Tr ceils and CD_4~+ CD_(25)~+ T ce]ls were extracted from peripheral blood lymphocytes by the microbeads separation.The purity of CD_4~+ CD_(25)~+ Tr cells and CD_4~+ CD_(25)~+ T cells was measured by flow cytometry.The growth inhibitory effect of CD_4~+ CD_(25)~+ Tr cells on CD_4~+ CD_(25)~+ T cells was assessed in vitro.Results The proportion of CD_4~+ CD_(25)~+ Tr cells was decreased significantly in URSA women(6.9?1.8)% than that in normal non-pregnancy women[(10.8?1.1)%] (P0.05).Conclusion The results suggest that decrease in proportion and function of CD_4~+ CD_(25)~+ Tr cells may be associated with URSA.

Objective To evaluate the left ventricle systolic function in patients with aortic stenosis (AS) underwent transcatheter aortic valve implantation(TAVI) by speckle tracking imaging and to observe the indicators in bicuspid aortic valves(BAV) and tricuspid aortic valves(TAV) groups.Methods Twenty nine patients with AS were enrolled,all of them underwent TAVI successfully.The regular echocardiography and 3D full-volume images were acquired on before and 3 days,1 month after TAVI.Longitudinal strain,circumferential strain,and three-dimensional left ventricle ejection fraction(3D-LVEF) were analyzed using Qlab software.Results Compared with the baseline,aortic valve blood flow velocity (AV),maximum aortic valve pressure gradient (AVPG-max),mean aortic valve pressure gradient (AVPG-mean),aortic valve area(AVA) after TAVI were improved significantly.Global longitudinal strain(GLS) had a improvement on 3 days after TAVI(all P ＜0.001),and further increased during 1 month after TAVI (all P ＜0.001).Global circumferential strain(GCS) were increased during 1 month after TAVI(all P ＜ 0.001).The 3D-LVEF after 1 month were improved significantly(all P ＜0.001).The BAV patients and TAV patients had similar changes in all of indicators observed.Conclusions The left ventricle systolic function has early improvement after TAVI,and further recovery during follow-up.The BAV patients can obtain a benefit from TAVI equally to the TAV patients.

Poxviruses, a type of ds-DNA viruses which mainly target at the epithelial cell, are the pathogens of human and animals. During the revolution of poxviruses, the viruses encode multiple proteins that regulate the immune system to monitor the viral reproductive cycle in host cells. The nuclear kappa B (NF-kappaB) pathway is essential to signal transcription in the innate immune system. Therefore, poxviruses have adopted different strategies to elude immune detection and destruction regulated by NF-kappaB. Further research in this field would help us develop preventive and therapeutic preparation for pox. Given the renewed interest in poxvirus, we review the current understanding of how the various classes of poxviralimmunomodulatory proteins target and manipulate the NF-kappaB pathway.
Animals ; Host Specificity ; Humans ; NF-kappa B ; metabolism ; Poxviridae ; physiology ; Signal Transduction

Objective To investigate the effect on adhesion and motion capbilities of adenoid cystic carcinoma(ACC) , by detecting the expression of nerve growth factor (NGF) , E-cadherin (E-cad) and S100A4 and the clinical significance in ACC tissues and analyzing the relationships between them with perineural invasion(PNI). Methods The expression of NGF, E-cad and S100A4 in ACC was detected with the way of immunohistochemistry SP, and then analyzing the expression level of them in different pathological types and histological regions in statistical ways on the basis of their relation with clinical and pathological parameters. Results The expression of NGF and S100A4 in PNI group [ 88% (23/26) and 77% (20/26) ] , was higher than that in NPNI group (8/16 and 7/16, P <0.05), and a positive correlation between them was identified in PNI group(r =0. 316,P <0.05). However the E-cad expression was lower in PNI group[31%(8/26) ,P<0. 05]. On the other hand it suggested a negative correlation with NGF (r = 0.385, P<0.05) as well as that with S100A4(r = -0.612,P =0.000). The expression level of NGF in fasciculus[ 83% (25/30)] has significant deviation compared with it in distant tumor tissues [ 47% (14/30), P<0. 05]. Conclusions In the PNI process of ACC, NGF plays important parts but not the only factor. It can increase the expression and activity of S100A4 but decrease E-cad expression through binding with its receptor. Thus, adhesion abilities of tumor cells was weakened and motional abilities was strengthen.

OBJECTIVETo study the effects of intestinal microflora alteration on specific and nonspecific immune function and hematopoietic function of mice.

METHODSSixty BALB/C mice were divided at random into two groups, experimental group and control group, with 30 mice in each. The mice in the experimental group were given kanamycin 50 mg while those in the control group were given distilled water intragastrically everyday for consecutive 10 days. After the 10 day treatment all the mice were sacrificed, and the cecal contents were collected for quantitative analysis of the intestinal bacterial flora. Certain indexes of immune function, including phagocytosis rate of macrophages, number of T lymphocytes positively stained by esterase and serum interleukin 2 (IL-2) content, and the weight of the spleen, granulocyte-macrophage colony stimulating factor etc. as indexes of hematopoietic function were determined.

RESULTSIn the group, the quantity of Enterobacteriaceae, Enterococcus, Bifidobacterium and Lactobacillus were significantly lower than that in the control group (P < 0.01). The number of PFC (plaque forming cells), the phagocytosis rate of macrophage, the number of T lymphocytes with positive NANE staining, the level of IL-2 significantly decreased when compared with that in the control group (P < 0.01). The weight of the spleen in the experimental group decreased when compared with that in the control group (P < 0.01). Levels of IL-3, GM-CSF, the total number of WBC and the proportion of neutrophil remarkably decreased as compared to that in the control group (P < 0.01). Analysis of the correlations between normal microflora, immunologic and hematopoietic indexes showed that marked positive correlations between the quantity of Bifidobacteria and each immune index including the levels of IL-3 and GM-CSF. There was a positive correlation between IL-2 and IL-3, IL-2 and GM-CSF as well.

CONCLUSIONThe application of antibiotics may cause changes in the structure and quantity of intestinal microflora. The dysbacteriosis may decrease the immune function of organism. The dysbacteriosis may decrease the hemopoietic function. The dysbacteriosis, the decrease in immune and hematopoietic function may affect one another. The balance in microecosystem should be emphasized and antibiotics should be applied rationally to reduce the side effects such as dysbacteriosis.

Animals ; Anti-Bacterial Agents ; pharmacology ; Esterases ; biosynthesis ; Feces ; microbiology ; Granulocyte-Macrophage Colony-Stimulating Factor ; analysis ; Interleukin-2 ; blood ; Intestines ; drug effects ; microbiology ; Kanamycin ; pharmacology ; Macrophages ; drug effects ; physiology ; Mice ; Mice, Inbred BALB C ; Models, Animal ; Organ Size ; Phagocytosis ; drug effects ; Spleen ; drug effects ; pathology ; T-Lymphocytes ; drug effects ; metabolism