The role of methyl-CpG binding domain protein 2 (MBD2) in an ApoE-deficient mouse model of age-related macular degeneration (AMD) was investigated. Eight-week-old Mbd2/ApoE double deficient (Mbd2(-/-) ApoE(-/-)) mice (n=12, 24 eyes, experimental group) and MBD2 (wt) ApoE(-/-) mice (n=12, 24 eyes, control group) were fed on Western-type diet for 4 months. The mice were sacrificed, and total serum cholesterol levels were analyzed and Bruch's membrane (BM) of the eyes was removed for ultrastructural observation by transmission electron microscopy. Moreover, intercellular adhesion molecule 1 (ICAM-1) immunoreactivities were evaluated by fluorescence microscopy in sections of the eyes in both groups for further understanding the function mechanism of MBD2. There was no significant difference in the total serum cholesterol levels between control group and experimental group (P>0.05). Transmission electron microscopy revealed that AMD-like lesions, various vacuoles accumulated on BM, notable outer collagenous layer deposits and dilated basal infoldings of retinal pigment epithelium (RPE) were seen in both groups, and the BM in control group was significantly thickened as compared with experimental group (P<0.05). Fluorescence micrographs exhibited the expression of ICAM-1 in choroid was higher in control group than in experimental group. We are led to conclude that MBD2 gene knockout may lead to accumulation of more deposits on the BM and influence the pathogenesis of AMD via triggering endothelial activation and inflammatory response in choroid, improving microcirculation, and reducing lipid deposition so as to inhibit the development of AMD-like lesions. Our study helps to provide a new therapeutic approach for the clinical treatment of AMD.

Dilated cardiomyopathy (DCM) is the most frequent pattern of non-ischemic cardiomyopathy with poor prognosis and high mortality.Studies in recent years found that non-coding small RNA molecules (miRNA) were closely related to the clinical course of DCM.The present article made a review on the expression pattern of miRNA and development of DCM study.

Adult ; Benzene ; poisoning ; Chronic Disease ; Cytochrome P-450 CYP2E1 ; genetics ; Female ; Humans ; Leukocyte Count ; Leukopenia ; chemically induced ; Male ; Middle Aged ; Polymorphism, Genetic

The role of methyl-CpG binding domain protein 2 (MBD2) in an ApoE-deficient mouse model of age-related macular degeneration (AMD) was investigated. Eight-week-old Mbd2/ApoE double deficient (Mbd2(-/-) ApoE(-/-)) mice (n=12, 24 eyes, experimental group) and MBD2 (wt) ApoE(-/-) mice (n=12, 24 eyes, control group) were fed on Western-type diet for 4 months. The mice were sacrificed, and total serum cholesterol levels were analyzed and Bruch's membrane (BM) of the eyes was removed for ultrastructural observation by transmission electron microscopy. Moreover, intercellular adhesion molecule 1 (ICAM-1) immunoreactivities were evaluated by fluorescence microscopy in sections of the eyes in both groups for further understanding the function mechanism of MBD2. There was no significant difference in the total serum cholesterol levels between control group and experimental group (P>0.05). Transmission electron microscopy revealed that AMD-like lesions, various vacuoles accumulated on BM, notable outer collagenous layer deposits and dilated basal infoldings of retinal pigment epithelium (RPE) were seen in both groups, and the BM in control group was significantly thickened as compared with experimental group (P<0.05). Fluorescence micrographs exhibited the expression of ICAM-1 in choroid was higher in control group than in experimental group. We are led to conclude that MBD2 gene knockout may lead to accumulation of more deposits on the BM and influence the pathogenesis of AMD via triggering endothelial activation and inflammatory response in choroid, improving microcirculation, and reducing lipid deposition so as to inhibit the development of AMD-like lesions. Our study helps to provide a new therapeutic approach for the clinical treatment of AMD.
Animals ; Apolipoproteins E ; genetics ; metabolism ; Bruch Membrane ; metabolism ; ultrastructure ; Cholesterol ; blood ; Choroid ; metabolism ; ultrastructure ; DNA-Binding Proteins ; genetics ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Macular Degeneration ; blood ; genetics ; metabolism ; Male ; Mice ; Mice, Knockout ; Microscopy, Electron, Transmission ; Microscopy, Fluorescence ; Retinal Pigment Epithelium ; metabolism ; ultrastructure

OBJECTIVETo evaluate the effect of alpha-galactosidase A (Gla) deficiency on FV Leiden (FVL) associated thrombosis in vivo.

METHODSTo generate the mice carrying mutations in Gla and FVL and analyze the tissue fibrin deposition in organs and thrombosis.

RESULTSIn the presence of FVL, Gla deficiency greatly increased tissue fibrin deposition compared with that in wild-type [Gla(-/0) FV(Q/Q) vs. Gla(+/0) FV(Q/Q) = (0.24 +/- 0.07)% vs. (0.086 +/- 0.049)%, P < 0.0001; Gla(-/-) FV(Q/Q) vs. Gla(+/+) FV(Q/Q) = (0.32 +/- 0.03)% vs. (0.06 +/- 0.005)%, P < 0.05]. With Gla deficiency, the number of thrombi on organ sections in FVL mice was significantly increased [(Gla(-/-) FV(Q/Q) and Gla(-/0) FV(Q/Q)) vs. (Gla(+/+) FV(Q/Q) and Gla(+/0) FV(Q/Q)) = 1.9 +/- 0.7 vs. 0.3 +/- 0.1, P < 0.05].

CONCLUSIONSGla deficiency could be an important genetic modifier for the enhanced thrombosis associated with FVL.

Animals ; Fabry Disease ; genetics ; Factor V ; genetics ; Genotype ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Mutation ; Thrombosis ; genetics ; pathology

Cell Culture Techniques ; methods ; Fixatives ; Humans ; Immunohistochemistry ; methods ; Tissue Fixation ; methods

OBJECTIVETo rapidly and sensitively detect the four virulence-associated factors of Streptococcus suis, a multiplex PCR was developed.

METHODSIn the process of this reaction, four distinct DNA targets were amplified. One target was based on the serotype 2 (and 1/2) specific cps gene and the others were based on Streptococcus suis mrp, epf (epf*) and sly gene, encoding the MRP, EF(EF*) and Sly proteins of Streptococcus suis. 72 isolates, which including 48 strains of Streptococcus suis and 24 strains of negative control, and 49 clinical specimens were detected by the multiplex PCR assay.

RESULTSAll PCR products were detected by electrophoresis on 1.2% agarose gels. With the 48 Streptococcus suis strains, the positive detection rates of cps2+, mrp+, epf+, epf*+ and sly+ were 16/48, 14/48, 12/48, 3/48 and 26/48,respectively. The results were confirmed by bacteriological examination. There were no specific amplification products including 49 clinical specimens and 24 negative control strains.

CONCLUSIONThe results demonstrated that multiplex PCR was a highly specific and sensitive diagnostic tool for the detection of virulence-associated factors of streptococcus suis.

Bacterial Proteins ; genetics ; Polymerase Chain Reaction ; methods ; Streptococcus suis ; genetics ; pathogenicity ; Virulence Factors ; genetics

Objective To evaluate the effectiveness of prevention of dental caries in preschool children by family-based oral health management. Methods A total of 144 families with 3 years old children in Huangpu District were randomly divided equally into intervention group and control group.The intervention period was 2 years.Semi-annual oral health guidance was given to families in the intervention group(including oral examination, children′s deciduous teeth coating with fluoride, dental caries filling, oral health lectures for families, distribution of family oral health brochures, etc.)Families in the control group received annual oral examination.After the intervention, the parents completed their child′s oral health questionnaire. Results In the 2-year oral monitoring period, the incidence of dental caries in the intervention group and the control group was 9.72% and 22.22%, respectively within one year, and the difference between the two groups was statistically significant(χ²=4.19, P < 0.05).The incidence of dental caries was 12.50% and 36.11% within two years, and the difference between the two groups was also statistically significant(χ²=10.91, P < 0.01).Result of the questionnaire survey showed that children in the intervention group had better eating and oral health habits than the control group, and the differences between the two groups were statistically significant(P < 0.05). Conclusion Regular and professional family oral health management can effectively reduce the incidence of dental caries and improve the oral health in preschool children.

In view of the present situation that edible marine shellfishes are combinedly contaminated by different kinds of lipophilic toxins, common lipophilic shellfish toxins in marine shellfishes were simultaneously detected by liquid chromatography-tandem mass spectrometry, and the safety risk of commercial marine shellfish was evaluated using the risk assessment method based on combined contamination of various toxins. Under the optimum conditions, satisfactory recoveries (63. 3％ - 88. 8％ ), precision ( relative standard deviations RSD≤14. 5％ ) and sensitivity (limit of detection in the range of 0. 5-2. 7 ng / g) of the method were achieved for all the analytes. Among the 105 commercially available shellfish samples, 42. 86％ of the samples had at least a kind of toxin. The highest average content was 47. 6 μg / kg of DTX1, which was the most serious contaminant for marine shellfishes. The total Expose Risk Index (∑ERI) was calculated based on Tolerable Daily Intake (TDI) and Acute Reference Dose (ARfD) of each toxin to evaluate the safety risk of commercial marine shellfish. The results showed that the ratio of commercially available marine shellfish with safety risk was 19. 05％ and the food safety risk of scallop was the highest. In summary, a new method based on the combined contamination of lipophilic shellfish toxins was successfully developed for risk assessment of the commercial marine shellfish. The proposed method is more harsh compared with the European Food Safety Authority (EFSA) regulation and can make shellfish consumers better to avoid the risk of poisoning.

Objective To observe the expression of stromal cell-derived factor-1α(SDF-1α)and glial cell line-derived neurotrophic factor(GDNF)genes in bone marrow stromal cells(BMSCs)of rhesus monkey. Methods With gene recombination technique,SDF-1α and GDNF genes obtained from cDNA were subcloned into pBudce4.1 vector to get pBudCE4.1-SDF-1-GDNF, evaluated by restriction enzyme analysis and sequencing analysis.The pBudCE4.1-SDF-1α-GDNF was transfected into BMSCs with lipofectamine2000.After 48 h,the expression of SDF-1α and GDNF was measured by RT-PCIL,Western blotting and immunohistochemistry. Results Correct construction of pBudCE4.1-SDF-1α-GDNF was identified by enzyme restriction analysis and sequencing analysis.Western blotting and immunohistochemistry confirmed the expressions Of SDF-1α and GDNF genes in the transfected cells.The protein expressions of GDNF and SDF-1α in the GDNF and SDF-1α transfected BMSCs were 6 times higher than those in the negative control cells. Conclusion The pBudCE4.1-SDF-1α-GDNF can successfully express SDF-1α and GDNF in BMSCs of rhesus monkey,which may be used in gene therapy for Parkinson's disease.