Blast Injuries ; Deafness ; etiology ; Humans

Adult ; Aged ; Biomarkers ; blood ; Female ; Humans ; Laminin ; blood ; Male ; Middle Aged ; Occupational Exposure ; Risk Assessment ; Silicon Dioxide ; adverse effects ; Silicosis ; blood ; diagnosis ; Superoxide Dismutase ; blood ; Transforming Growth Factor beta ; blood

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Occupational Health ; Pneumoconiosis ; diagnostic imaging ; Radiographic Image Enhancement ; methods ; Young Adult

Animals ; Disease Progression ; Kidney Diseases ; physiopathology ; Kidney Glomerulus ; drug effects ; pathology ; Rats ; Renal Agents ; pharmacology ; Renin-Angiotensin System ; drug effects ; Sclerosis

Objective To rational design HCV DNA vaccine candidates and evaluate their specific immunity to HCV in mice. Methods We design to construct two DNA vaccine candidates, one consists of E_2 (the envelope glycoprotein 2 of HCV) gene only, the second consists of E_2-gAD (Globular Domain of Human Adiponectin) fusion gene via overlapping PCR. Confirm the expression of the DNA vaccines by Western blotting, and then vaccinated by injection of DNA vaccines with gene electrotransfer (GET) in BALB/c mice. The immune response was measured by IFN-gamma ELISPOT. Results The DNA vaccine candidate consists of E_2-gAD could effectively express in vitro , and it could induced a higher anti-HCV T cell response in mice than the one consists of E_2 only. Conclusion The HCV DNA vaccine consists of E_2-gAD fusion can increase the immunity of the E_2 to some extend, and the research paved a way to develop and optimize the novel HCV DNA vaccine.

OBJECTIVETo investigate the use value of picrosirius red staining plus polarized microscopy to observe the dynamic changes of collagen fiber in lung fibrosis in silicotic mice model.

METHODSThe experimental mice were divided into control and quartz groups. 0.2 g/kg weight of quartz was injected intratracheally in quartz group. Lung tissues were collected at the 1st, 3rd, 5th, 7th, 14th and 28th day after injection respectively. Lung tissue slides were stained with picrosirius red. With the aid of polarized microscope, image analysis software, the distribution and change of type I and type III collagen could be qualitatively and quantitatively analyzed. Lung tissue hydroxyproline was determined by chloramines T method.

RESULTSIn early stage the predominant increment was type III collagen, but in late stage type I was predominant. The contents of both type collagen tended to increase as postexposure time prolonged. The time course of the ratio of type I to type III showed increasing trend, and there was a statistical significance on day 28 (1.49 +/- 0.39 vs 0.59 +/- 0.24, P < 0.05). The total area of collagen was positively correlated with hydroxyproline concentration of lung tissue (r(2) = 0.928 5, P < 0.01).

CONCLUSIONPicrosirius red staining combined with polarized microscopy and digital image processing is a useful method to elucidate collagen accumulation, distribution and subtype ratio in silicosis.

Animals ; Azo Compounds ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Coloring Agents ; Hydroxyproline ; metabolism ; Lung ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred Strains ; Microscopy, Polarization ; Pulmonary Fibrosis ; chemically induced ; metabolism ; pathology ; Quartz ; toxicity ; Staining and Labeling

Chickenpox ; complications ; Child ; Hemiplegia ; diagnosis ; virology ; Humans

Objective To investigate the difference of amplitude of low-frequency fluctuation (ALFF) and fraction of amplitude of low-frequency fluctuation(fALFF) between Alzheimer's disease (AD)patients and normal aging (NA) controls by voxel-based analysis.Methods Thirty-one AD patients and 44 NA controls were enrolled in the study.Blood oxygen level dependent functional (BOLD) EPI data were obtained during resting-state by using 32-channel head coil.Data were realigned,normalized and then smoothed with 8 mm FWHM kernel.Resting-state fMRI toolkit(version 1.6) was used to generate ALFF and fALFF images.Independent two sample t-test was performed with SPM5 to compare ALFF and fALFF of AD and NA controls.Pearson correlation analysis was performed to examine the relationship between MMSE score and ALFF,fALFF parameters.The significance level was set to be uncorrected O.001 on the voxel level and 0.05 on the cluster level.Results AD patients showed increased ALFF in left temporal lobe (0.492 ±0.119) and right cingulated cortex (0.434 ± 0.093) of AD patients,which were 0.443 ± 0.068 and 0.380 ±0.081 in NA controls (t =2.658,2.227,P ＜ 0.05).Decreased fALFF was found in bilateral posterior cingulate cortices (1.167 ± 0.203) and increased fALFF was found in bilateral temporal lobes (left 1.226 ±0.127,right 1.146 ±0.214) with left side dominance,which were 1.453 ±0.269,1.134 ±0.088,1.014 ± O.132 in NA controls (t =5.001,3.695,3.285,P ＜ 0.05).Bilateral temporal ALFF and fALFF correlated with MMSE positively (r =0.768—0.909,P ＜ 0.05) with left dominance.Conclusion AD patients showed increased resting-state functional MRI changes correlated with MMSE score in the temporal lobes with left dominance,which indicated left temporal lobe may be the best location for the observation of disease progression in AD patients.

Objective To study the regulation of microRNA 199a (miR-199a) on adhesion,migration and invasion ability of human eutopic endometrial stromal cells (ESC) from patients with endometriosis.Methods ESC were transfected with miR-199a mimics or negative control (NC) RNA by lipofectamine 2000.The adhesion,migration and invasion ability of ESC were detected by cell adhesion assay,scratch assay,cell migration assay and matrigel invasion assay,respectively.Luciferase reporter assay was used to evaluate whether IKKβ was the target gene of miR-199a.The expression of ikappa B kinase beta (IKKβ),inhibitory kappa B alpha (IκB-α),phospho-IκB-α (p-IκB-α) and nuclear factor-kappa B (NF κB) protein were measured by western blot.Results ( 1 ) Adhesion potential:the adhesion inhibitory rates were ( 14 ± 4 )％ in miR-199a group and 0 in control group,which showed significant difference (P＜0.01 ).(2) Migration and invasion:in the scratch assay,ESC transfected with miR-199a exhibited a lower scratch closure rate than that of controls.In migration and invasion assays,the migration and invasion ability of miR-199a group were significantly decreased compared with those of NC group [ 130 ± 31 vs.247±36 (P＜0.01); 63 ± 15 vs.133 ± 17 (P＜0.01),respectively].(3) The luciferase activity of miR-199a group was significantly lowered than that of control group [ 0.160 ± 0.006 vs.0.383 ± 0.083 ( P ＜0.01 ) ].The protein levels of IKKβ,p-IκB-α,IκB-α and NF-κB of 0.350 ±0.195,0.443 ±0.076,1.970 ±0.486 and 0.454 ± 0.147 in miR-199a group were significantly different compared with the NC group in which the protein levels were set at 1.000 ( P ＜ 0.01 ).Conclusions miR-199a can inhibit the adhesion,migration and invasion of the ESC.IKKβ is the target gene of miR-199a in ESC.One of the mechanisms of the inhibition effect is probably that miR-199a inhibits the activation of NF-κB signaling pathway by targeting IKKβ gene.

Objective To study the effects of fluoride on minichromosone maintenance(MCM)3 mRNA and the bone formation-related gene:bone sialoprotein(BSP),osteocalcin(OC),osteopontin(OP)mRNA expression on human osteoblast cells.The expression of MCM3 was tested for diagnosis and surveillance value on osteoblast treated with excess fluoride.Methods Human osteoblast cell(Saos-2)was cultured in McCoy5A medium and treated with fluoride(sodium fluoride,NaF).There were eight groups including:0(control),0.625,1.250,2.500,5.000,10.000,20.000,40.000 mg/L groups.Expression of MCM3,BSP,OC,OP mRNA were detected by real-time PCR.Dual-standard curve method was used for analysis.ALPase was determined by measuring the absorbance using a micro titer plate reader. Results Expression of MCM3 mRNA was lower in the 0.625,1.250,2.500,5.000,20.000, 40.000 mg/L groups(0.059 ± 0.003,0.027 ± 0.001,0.272 ± 0.004,0.115 ± 0.002,0.137 ± 0.004,0.754 ±0.002, all P > 0.05) and was higher in10.000 mg/L group(21.300 ± 1.200, P < 0.01 ) than control group( 1.000 ±0.020), especially 10.000 mg/L group was higher than groups treated with fluoride(all P < 0.01 ), the differences among groups were significant(F = 305.842, P < 0.01 ). Expression of BSP mRNA was significantly higher in 0.625,1.250,2.500,5.000,10.000 mg/L groups(71.80 ± 3.60,133.00 ± 7.20,85.50 ± 0.60,80.90 ± 1.20,304.00 ± 21.00)than the control group( 1.00 ± 0.04), especially 10.000 mg/L group was higher than others groups treated with fluoride(all P < 0.01 ), the differences among groups were signifieant(F = 159.531, P < 0.01 ). Expressions of OC mRNA were higher in 0.625,1.250,2.500,5.000 mg/L groups(110.00 ± 12.00,143.00 ± 2.10,90.60 ± 4.10,23.70±1.20) than control group(1.00 ± 0.01, all P < 0.01), and the differences among groups were significant (F = 158.734, P < 0.01 ). Expression of OP mRNA were higher in 0.625,1.250,2.500,5.000,10.000,20.000 mg/L groups(167.00 ± 11.20, 111.00 ± 12.10,72.50 ± 3.50,134.00 ± 14.00,42.30 ± 2.40,45.20 ± 3.30) than the control group(1.00 ± 0.04, all P < 0.05 or < 0.01 ), the differences among groups were significant(F = 60.226, P < 0.01 ).Compared with control group(4.2 ± 1.2), the ALPase activity was increased in all groups treated with fluoride (6.0 ± 0.4,5.8 ± 0.1,5.7 ± 0.4,7.7 ± 1.1,19.2 ± 2.4,8.5 ± 3.0,18.1 ± 4.2), but only 10.000 mg/L and 40.000 mg/L groups were higher than control group and other groups treated with fluoride(all P < 0.01 ), the differences among groups were signifieant(F = 7.806, P < 0.01 ). Conclusions Irregular expression of MCM3 mRNA is not suitable as a diagnostic and monitoring biomarker of osteoblasts exposed to excessive fluoride. Fluoride may affect the osteoblast-related gene expression and to promote osteogenic differentiation.