Objective To investigate the effects of staphylococcal enterotoxin A SEA gene on target cells mediated by replicative-deficient recombinant adenovirus vector. Methods Lymphocytes of C57BL/6 mice were infected with various titers of recombinant adenoviruses. Supernatants were collected after 12 h, 24 h, 48 h, 72 h, 96 h, 120 h and 144 h of incubation and analyzed for proliferation of lymphocytes by MTT assay. IL-2 level in the culture supernatants was measured with ELISA. The killing effect of lymphocytes was also observed by MTT assay. Results Proliferation response and elevated levels of IL-2 were observed in experimental group. The killing effect on B16 cells was stronger in experimental group, which seemed to be dose-dependent with the increase of ratio of lymphocytes/target cells. Conclusions SEA gene can be expressed in lymphocytes of C57BL/6 mice mediated by replicative-deficient recombinant adenovirus vector. The expressing products can activate lymphocytes of C57BL/6 mice, which kill B16 cells in vitro.

Objective To probe into the application of left atrial volume tracking technique(LAVT)on the evaluation of right atrial function in patients with pulmonary hypertension.Methods Forty-one patients with pulmonary hypertension (pulmonary hypertension group) and 37 control subjects (control group) were involved.Right atrial maximal volume (RAVmax),right atrial presystolic volume(RAVpre),right atrial minimal volume (RAVmax),systolic right atrial filling rate (dv/dtS),early diastolic right atrial emptying rate(dv/dtE) and late diastolic right atrial emptying rate(dv/dtA) was derived by LAVT.Right atrial passive emptying volume (RAVp),right atrial passive emptying fraction (RAVpEF),right atrial active emptying volume (RAVa),right atrial active emptying fraction (RAVaEF),right atrial total emptying volume (RAVt)and right atrial total emptying fraction (RAVtEF) was calculated.All the right atrial volume parameter was corrected by body surface area to obtain right atrial volume index (RAVI).Results RAVImax,RAVImin,RAVIpre,RAVIt,RAVIa,dv/dtS and dv/dtA in pulmonary hypertension group was higher than that in control group [(78.39 ± 49.35) ml/m2 vs.(24.80 ± 11.91) ml/m2,(62.59 ± 46.56) ml/m2vs.(17.46 ± 8.40)ml/m2,(70.12 ± 48.03) ml/m2 vs.(20.02 ± 9.46) ml/m2,(18.77 ± 11.47) ml/m2 vs.(9.35 ± 6.74) ml/m2,(8.53 ± 9.81) ml/m2 vs.(3.25 ± 3.00) ml/m2,(145.85 ± 80.56) ml/s vs.(86.44 ± 48.46) ml/s,(155.63 ±126.47) ml/s vs.(67.74 ± 33.27) ml/s],and RAVIp in pulmonary hypertension group was lower than that in control group [(6.09 ± 5.16) ml/m2 vs.(10.23 ± 11.12) ml/m2],and there were significant differences (P ＜0.05).But there were no significant differences in RAVItEF,RAVIpEF,RAVIaEF and dv/dtE between two groups (P＞0.05).Conclusions In patients with pulmonary hypertension,right atrial booster pump function and reservoir function increases,while right atrial conduit function decreases.LAVT has a potential ability to evaluate right atrial function.

Recepteur d′origine Nantais(RON),a tyrosine kinase receptor ,is a growth factor receptor belonging to the proto-oncogene met family and has been proved to display abnormal expres?sion in many types of tumors. The RON receptor is activated by binding to the ligand macrophage stim?ulating protein,overexpression of the receptor,variants of the proto-oncogene and by point mutations of the kinase region. The downstream transduction of RON by mitogen-activated protein kinases and phosphoinositide3-kinase signaling pathways can help regulate the proliferation,apoptosis,migration and invasion of tumor cells. A better understanding of the mechanisms and related signaling pathways of RON activation in tumor progress and development will provide more information for the RON-based target therapy.

T mutation was detected in the 4th propositus at the 9th intron,but any COL1A1 or COL1A2 gene mutation was detected in the third propositus and the other members in the former families.Conclusions The genetic mutation of COL1A1 may result in OI in China,but other mutations may also exist.Moreover,the phenotype was influenced not only by OI genotype,but also by the genetic background,environment and other factors.

Objective　To analyze the effects of flexible ureteroscopic Holmium laser lithotripsy on renal function in the treatmen of renal calculi.　Methods　From October 2010 to March 2012,41 cases of solitary renal calculi (24 males and 17 females) were treated with flexible ureteroscopic Holmium laser lithotripsy ( FURL).Patient's mean age was 51.9 ± 15.9 years ( range from 31 to 88 years).Locations of renal calculi detected by image study were 22 cases in middle and upper calyx,9 cases in lower calyx,10 case in renal pelvis.The mean size of calculi was 16.9 ±6.0 mm (range from 10 to 28 mm).Blood samples (2 h pre-operatively,and then 2 h,12 h,24 h,48 h,72 h post-operatively) were collected and serum　NGAL,serum Cys-C were tested　Results　The measured levels of pre-operative NGAL and Cys-C were 3.5 ± 0.6μg/L,501.7±121.3 μg/L,and levels of post-operative NGAL at 2 h,12 h,24 h,48 h and 72 h were 4.2±0.8 μg/L,5.0±1.0 μg/L,4.9±1.4 μg/L,4.3± 1.1 μg/L and 3.8 ±0.1 μg/L,while the according levels of Cys-C were ( 516.4 ± 126.2 ) μg/L,( 723.8 ± 134.8 ) μg/L,( 770.4 ± 162.8 ) μg/L,(671.7 ± 138.3 ) μg/L and 574.0 ± 116.7 μg/L.Serum NGAL began to increase 2 hours after operation (P ＜0.05,and reached the peak in 12 hours,it began to decline 12 hours after surgery,but it was still higher at 72 hours than pre-operative level (P ＜ 0.05.Serum Cys-C showed no obvious ascent 2 hours after operation in FURL group ( P ＞ 0.05 ),but increased obviously 12 hours after operation and lowered down after the peak that occurred 24 hours after surgery.Serum Cys-C still remain above the baseline 72 hours after operation (P ＜0.05).　Conclusions　Flexible ureteroscope lithotripsy can cause reversible damage to renal function after surgery.

Objective:To study the inhibitory effect of siRNA on Heparanase (HPA) expression in SKOV3 cells. Methods:Two pairs of 21 bp reverse repeated sequence targeting HPA RNA (spaced by 9 bp nueleotide) were synthe- sized and were cloned into plasmid PGenesil-1 to construct recombinant plasmid PGenesil-1(+)-HPA expressing 2 hair- pin siRNAs.The inhibition of HPA gene was detected by RT-PCR,real-time PCR and immunohistochemical staining after PGenesil-1(+)-HPA was stably transfected into SKOV3 cells.Results:The recombinant plasmid PGenesil-1(+)-HPA (expressing 2 hairpin siRNAs) was successfully constructed.RT-PCR,real-time PCR and immunohistochemical staining showed that there was a significant decrease in HPA mRNA and protein level in experimental group compared with those in control group.Conclusion:siRNA targeting HPA mRNA can specically suppress the expression of HPA gene in SKOV3 cells;RNA interference method provides a new way for studying the role of HPA and gene therapy of cancer.

Objective:To study the cytocompatibility of Zr-Cu-Al-Ag alloy coated by micro-arc oxidation.Methods:Components of Zr-Cu-Al-Ag alloy coated by micro-arc oxidation in three different voltages of 300 V,350 V and 400 V,Zr-Cu-Al-Ag alloy as cast condition and TI6Al4V alloy were made for the test.The water extracted from the components were obtained according to national standard.The L929 cells were cultivated in vitro in the extracts of these components separately.The L929 cells,cultured in Dulbecco's modified Eagle medium supplemented with 10 ％ fetal calf serum,served as the negative control group.And cells,cultured in Dulbecco's modified Eagle medium supplemented with 10 ％ fetal calf serum and 64 g/L phenol,served as the positive control group.The cytocompatibility of these components were evaluated by MTT colorimetric.Results:The cytotoxicity of Zr-Cu-Al-Ag alloy coated by micro-arc oxidation is 0 grade.Microscopy showed that the morphology of L929 cells,cultured in the extracts of Zr-Cu-Al-Ag alloy coated by micro-arc oxidation were normal.There were no significant differences between micro-arc oxidationt and negative control groups.The cell multiplication curves of micro-arc oxidation and negative control groups were nearly overlapping and in the linearity increasing trend.The OD in micro-arc oxidation groups had no significant differences with negative control group (P＞0.05),but were higher than that of Zr-Cu-Al-Ag alloy as cast condition,TI6Al4V alloy and positive control groups (P＜0.05).Conclusions:The cytocompatibility of Zr-Cu-Al-Ag alloy has been improved by micro-arc oxidation technique.

BackgroundThe three-dimensional configuration of the nasolacrimal canal is highly variable with age,gender,and race.But enlargement of the nasolacrimal canal has sparsely been reported in the literature.Objective Computed tomography dacryocystography was performed in patients with unilateral congenital nasolacrimal duct obstruction and normal children to analyze the difference of bilateral nasolacrimal canal.MethodsThis is a retrospective study.Axial scanwith sagittalandcoronalreconstructionwas appliedin computedtomography dacryocystography.Diameters of bilateral nasolacrimal canal of 20 unilateral congenital nasolacrimal duct obstruction patients and 20 normal children were measured.Written informed consent was obtained from each child ' s parents before examination.ResultsThe lacrimal sac,nasolacrimal duct and the peripheral tissue were clearly exhibited by computed tomography dacryocystography.The diameters of the origination,the middle part and the distal end of affected nasolacrimal duct were(5.5±1.4),(5.3±1.2),(5.3±1.6) mm,and normal ones were(3.9±0.8 ),(3.5± 0.8 ),( 3.9± 1.3 ) mm,respectively.These results were statistically significant ( t =5.200,6.967,2.932,P＜ 0.05 ).There was no statistically significant difference in bilateral nasolacrimal canal of normal children (t =0.346,0.281,0.312,P＞0.05 ).Conclusions Computed tomography dacryocystography can image lacrimal passage and their peripheral tissues clearly.The affected nasolacrimal canal diameters of unilateral congenital nasolacrimal duct obstruction were much larger than the fellow sides.The pathogenesis of this phenomenon need much research.

Objective In order to study the biological characteristics of macrophages and provide the materials to study the survival mechanism of intracellular parasites, we conducted this study to establish a high-purity alveolar macrophage isolation and culture method.Methods Goat lungs were lavaged with normal saline in sterile environment several times, and cells were collected and then goat alveolar macrophages were purified by density gradient centrifugation using peripheral blood mononuclear cells (PBMC) solution.The isolated goat alveolar macrophages were cultured in cell culture medium containing 10% fetal bovine serum and cell morphology was observed under an inverted microscope every day,and the phagocytic activity of the cells was detected by chicken red blood cell phagocytosis test.Flow cytometry was used to detect CD14, a characteristic monocyte-macrophage surface marker.Results The adherent cells were characterized by typical macrophage morphology, pseudopodia and protrusions, showing round and irregular shape, rich cytoplasm, and large cell body.Of the cultured macrophages, 54.5% could phagocytize chicken erythrocytes and showed good phagocytic activity.After one month of in vitro culture, 93.7% of the cells were able to express CD14 antigen, which had a macrophage-specific immunophenotype.Conclusions The alveolar macrophages obtained in this study have high purity and good bioactivity, thus provide a cell model for studying the immune mechanism of intracellular parasites.

ObjectiveTo explore the effects of heat treatment and ultraviolet B (UVB) radiation alone or in combination on the expression of heat shock protein (HSP) 72 in human epidermal melanocytes.Methods Melanocytes were obtained from human foreskin,and subjected to primary culture.After 3 to 5 passages,the melanocytes were classified into 4 groups:control group (receiving no treatment),heat treatment group (treated with heat at 42 ℃ for 1 hour every day for 3 days),UVB group(irradiated with UVB at 50 mJ/cm2 daily for 3days),combination group(treated with heat at 42 ℃ for 1 hour followed by irradiation with UVB at 50 mJ/cm2daily for 3 days).After another 2- to 6-hour culture following the last treatment,melanocytes were collected and subjected to real time PCR and Western blot for the detection of HSP72 mRNA and protein expression,respectively.ResultsThe mRNA and protein expressions of HSP72 were significantly higher in the heat treatment group and combination group than in the control group (mRNA:6.584 ± 0.871 and 7.269 ± 0.454 vs.0.975 ± 0.089,both P ＜ 0.001; protein:2.022 ± 0.058 and 2.080 ± 0.045 vs.0.532 ± 0.033,both P ＜ 0.001 ),but was similar between the UVB group and control group (mRNA:0.832 ± 0.084 vs.0.975 ± 0.089,P ＞ 0.05;protein:0.546±0.021 vs.0.532 ± 0.033,P ＞ 0.05).The ANOVA of factorial design showed that neither heat treatment nor UVB irradiation had interaction effect on the mRNA or protein expression of HSP72 (F =2.106,1.399 respectively,both P ＜ 0.05).ConclusionsHeat treatment can cause an increase in the expression of HSP72,which may enhance the function of melanocytes and protect melanocytes from UVB induced damage.