Forensic Medicine ; Humans ; Kidney Transplantation ; Polycystic Kidney Diseases

Adolescent ; Adult ; Cell Transplantation ; Child ; Epidermis ; cytology ; Female ; Humans ; Male ; Suspensions ; Transplantation, Autologous ; Vitiligo ; therapy

Small ubiquitin-related modifiers (SUMOs) belong to an important class of ubiquitin like proteins. SUMOylation is a post-translational modification process that regulates the functional properties of many proteins, among which are several proteins implicated in neurodegenerative diseases. This study was aimed to investigate the changes of SUMO-1 expression and modification, and the relationship between SUMO-1 and Alzheimer's disease (AD) pathology in APP/PS1 transgenic AD mice. Using Western blot, co-immunoprecipitation and immunofluorescent staining methods, the SUMO-1 expression and modification and its relation to tau, amyloid precursor protein (APP) and β-amyloid protein (Aβ) in the 12-month-old APP/PS1 transgenic AD mice were analyzed. The results showed that: (1) Compared with the normal wild-type mice, the expression and modification of SUMO-1 increased in brain of AD mice, which was accompanied by an increase of ubiquitination; (2) In RIPA soluble protein fraction of cerebral cortex, co-immunoprecipitation analysis showed tau SUMOylated by SUMO-1 increased in AD mice, however, AT8 antibody labeled phosphorylated tau was less SUMOylated whereas PS422 antibody labeled phosphorylated tau was similar to control mice; (3) Double immunofluorescent staining showed that SUMO-1 could distributed in amyloid plaques, appearing that some of SUMO-1 diffused in centre of some plaques and some of SUMO-1 co-localized with AT8 labeled phosphorylated tau forming punctate aggregates around amyloid plaques which was concerned as dystrophic neurites, however, less Aβ, APP and PS422 labeled phosphorylated tau were found co-localized with SUMO-1. These results suggest that SUMO-1 expression and modification increase abnormally in transgenic AD mice, which may participate in modulation of the formation of senile plaques and dystrophic neurites.
Alzheimer Disease ; physiopathology ; Amyloid beta-Peptides ; metabolism ; Amyloid beta-Protein Precursor ; metabolism ; Animals ; Brain ; pathology ; Mice ; Mice, Transgenic ; Neurites ; pathology ; Phosphorylation ; Plaque, Amyloid ; physiopathology ; SUMO-1 Protein ; metabolism ; Sumoylation ; tau Proteins ; metabolism

Objective To affirm and calssify the arsas with high water iodine and the endemic areas of indline excess goiter in Jiangsu Privince accirding to mational standard.Methods A cross section survey was condueted in 2005 at township level in Fenxian,Tongshan,Suining,Pizhou counties in Xuzhou municipal and Chuzhou district in Huai'an municipal in Jiangsu Province One sample of dringing well water from five directions in the five villages located dircetions of every township namely east west south north and central.was tested for its water iodine concentration.If the sample number sas less than 5 in one village,then all the well water would be tested Endemic stutas of iodine excess goiter was investigated in those townships whose menian water iodine comcentration was between 150 to 300 μg/L Then status was affirmed and reclassifed according.to National Criteria GB/T 19280-2003 Results In all 158 tawnships from the 6 counties the median of water iodine concentration in 79 townships were over 150 μg/L with 32 townships in the range of 150 to 300μg/L In those 32 townships 9 met the criteria of area of high water iodine 23 accorded with that of encemic areas of iosine excess goiter 16 had stopped the supply of non-iodzed salt in advace,but 16 newly detected areas were still served with iodized salt Four were 300 μg/L Conctusions Iodized salt intervention should be stopped in all townships with the problems of high water iodine and the endmic areas of iodine excess goter in order to prevent the possible hazards due to double intade.

OBJECTIVEElectron microscopical study of infected cells to identify the pathogenic agent of SARS.

METHODSVero E6 cells infected with lung autopsy samples or nasopharyngeal swabs from SARS patients of Beijing and Guangzhou were inoculated. The supernatant and cultured cells exhibiting identifiable cytopathic effect (CPE) were prepared for electron microscopic study.

RESULTSExamination of CPE cells on thin-section revealed characteristic coronavirus particles within the cisternae of endoplasmic reticulum, Golgi apparatus, vesicles and extracellular space. They were mainly spherical or oval in shape, annular or dense, about 80 nm in diameter. Negative-stain electron microscopy identified coronavirus particles in culture supernatant, 80 - 120 nm in diameter, with club-shaped surface projections. Elongated, rod-, kidney- or other irregular shaped virons with the size of 100 - 200 nm by 60 - 90 nm were also found in the cultured cells infected with the lung samples from the Guangdong patients. Infectious virons entered cells by endocytosis or membrane fusion and released through a budding process.

CONCLUSIONThese data indicate a novel coronavirus as the causative agent of SARS. Most viral particles showed typical characteristics of coronavirus. The potential role of special shape viruses is expected to be further investigated.

Animals ; Cercopithecus aethiops ; Humans ; Microscopy, Electron ; SARS Virus ; ultrastructure ; Severe Acute Respiratory Syndrome ; virology ; Vero Cells

OBJECTIVETo establish a method for gene delivery in murine renal tissue using lentivirus vector encoding miR-483-5p.

METHODSThirty-five C57BL/6J mice were randomly divided into control group, low-dose treatment group (5 µL each kidney) , and high?dose treatment group (20 µL each kidney), and in the latter two groups, the lentivirus vector encoding miR-483-5p were injected in the renal cortex. The tissue samples were collected at 7 and 21 days after the injection. A transgenic mouse model with inducible systemic overexpression of miR-483-5p was established in TG483 mice. The Cre-loxp system was used to create a mouse model with renal tubule-specific expression of miR-483-5p. The levels of BUN in the mice were detected and HE staining and fluorometric TUNEL assay were used to observe the morphological changes of the kidneys; real-time qPCR was used to detect miR-483-5p expression in the renal cortex.

RESULTSThe mice with overexpression of miR-483-5p had normal renal function without obvious pathological changes or apoptosis in the renal tissue. Renal cortex injection of 20 µL lentivirus resulted in obviously increased level of miR-483-5p at 21 days (1.2∓0.43 vs 8.6∓1.09, P<0.001). miR-483-5p showed a low expression (0.9∓0.09 vs 1.7∓0.19, P<0.05) in TG483 mice and a high expression in the kidney of the transgenic mice established using the Cre-loxp system (1.6∓1.13 vs 12.36∓3.89, P<0.05).

CONCLUSIONThe transgenic mice with renal tubule-specific expression of miR-483-5p show normal renal function, and this model facilitates further study of the role of miR-483-5p in the kidney.

OBJECTIVETo investigate the effect of estradiol on the expression of antioxidant enzymes in osteoblasts and its role in postmenopausal osteoporosis.

METHODSRat models of osteoporosis established by ovariectomy were treated with estradiol for 3 months, and the changes in serum levels of reactive oxygen species (HO) and antioxidant enzymes (γ -GCS, GSH-ST and GSH-px) were detected. The effects of estradiol on the expression of γ -GCS mRNA and protein in osteoblast-like cells MC3T3-E1, MG63 and OB were examined with PCR and Western blotting. Using a mRNA microarray, we analyzed the changes in the expressions of 84 antioxidant enzymes in the osteoblast cell line MC3T3-E1 following estradiol treatment, and the enzymes with significant changes were verified by PCR. CCK-8 kit was used to evaluate the effect of estradiol and antioxidant NAC on the proliferation of MC3T3-E1 cells.

RESULTSRat models of osteoporosis were successfully established with ovariectomy. The osteoporotic rats showed significantly increased serum level of reactive oxygen species (H2O2) and decreased levels of antioxidant enzymes. Estrogen treatment of the osteoporotic rats obviously reversed the phenotype of osteoporosis, lowered serum level of reactive oxygen species, and increased the level of γ -GCS. In MC3T3-E1, MG63 and OB cells, estradiol treatment significantly upregulated the expression levels of γ -GCS mRNA and protein. In MC3T3-E1 cells treated with estrogen, the mRNA chip identified 6 upregulated antioxidant enzymes (Gpx6, Gstk1, Nos2, Prdx2, Ngb and Ccs), and the results of PCR verified that estradiol upregulated Ccs and Ngb mRNAs in MC3T3-E1, MG63 and OB cells. Estradiol and antioxidant NAC obviously promoted the proliferation of MC3T3-E1 cells.

CONCLUSIONEstradiol significantly increases the expression of antioxidase γ -Gcs, Ccs and Ngb in osteoblasts in vitro. Postmenopausal osteoporosis is closely related with the increase of reactive oxygen species and the decrease of antioxidant levels. In osteoblasts, estrogen deficiency may increase the level of reactive oxygen species, decrease the level of antioxidant enzymes, activate the oxidative stress cascade, and consequently inhibit the proliferation of osteoblasts to aggravate the condition of osteoporosis.

OBJECTIVETo investigate the effect of birth weight and early growth on body fat composition and insulin sensitivity.

METHODSThe birth and growth data of 258 children of 6 to 7 years old in Guangzhou were collected from Jun.2009 to Feb. 2010. Physical and laboratory examination were preformed, which included body weight, body height and body fat composition index (body mass index (BMI), percentage of body fat (PBF), waist circumference to height ratio (WtHR), etc). Fasting blood glucose and insulin were measured. The homeostasis model assessment model for insulin resistance index (HOMA-IR) was calculated. According to birth weight, the children were divided into three groups from light to heavy: BW-I, BW-II, BW-III group. Then according to change in weight SDS between 0 and 36 months, the children were divided into three groups: changers up (CU), non-changers (NC), changers down (CD) group. The effect of birth weight and early growth on body fat composition and insulin sensitivity were analyzed.

RESULTSChange in weight SDS between 0 and 36 months was higher in BW-I group (1.06 ± 1.29) than in the BW-II group (-0.19 ± 0.94) and BW-III group (-0.10 ± 1.20) (all P values < 0.01). Birth weight of the CU group ((2.90 ± 0.47) kg) was lower than that of the NC group ((3.22 ± 0.34) kg) and the CD group ((3.57 ± 0.37) kg) (all P values < 0.01). The body fat composition index of BMI, PBF and WtHR were higher in the BW-III group ((16.35 ± 2.13) kg/m(2), (17.03 ± 5.88)%, (0.479 ± 0.033)) than in the BW-I group ((15.46 ± 2.06) kg/m(2), (14.06 ± 5.25)%, (0.459 ± 0.032)) and BW-II group ((15.47 ± 1.58) kg/m(2), (14.09 ± 5.01)%, (0.460 ± 0.025)) (P < 0.01), while there was no significant difference between the BW-I group and the BW-II group (P > 0.05). The body fat composition index of BMI, PBF and WtHR were higher in the CU group ((16.44 ± 2.20) kg/m(2), (16.51 ± 5.78)%, (0.473 ± 0.034)) than in the NC group ((15.62 ± 1.74) kg/m(2), (14.49 ± 5.30)%, (0.463 ± 0.030)) and the CD group ((15.26 ± 1.85) kg/m(2), (14.24 ± 5.54)%, (0.462 ± 0.031)) (all P values < 0.05). In the CU group, BMI, PBF and WtHR were higher in the BW-III-CU group ((18.76 ± 2.56) kg/m(2), (22.19 ± 8.28)%, (0.512 ± 0.029)) than in the BW-I-CU group ((16.04 ± 2.14) kg/m(2), (15.54 ± 5.28)%, (0.467 ± 0.034)) and BW-II-CU group ((16.70 ± 1.36) kg/m(2), (17.12 ± 4.44)%, (0.474 ± 0.017)) (all P values < 0.05), while there was no significant difference between the BW-I-CU group and the BW-II-CU group (P > 0.05). HOMA-IR was higher in the CU group (1.27 ± 0.44) than in the NC group (1.08 ± 0.31) and the CD group (1.00 ± 0.36) (all P values < 0.01). In the CU group, HOMA-IR was higher in the BW-III-CU group (1.69 ± 0.48) than in the BW-I-CU group (1.21 ± 0.41) and the BW-II-CU group (1.27 ± 0.44) (all P values < 0.01), while there was no significant difference between the BW-I-CU and BW-II-CU group (P > 0.05).

CONCLUSIONAccording to birth weight tertile, both lower birth weight individuals with more weight change-up growth postnatal early and higher birth weight individuals had greater body fat composition in childhood. They were high-risk people of insulin resistance.

Birth Weight ; Body Composition ; Body Mass Index ; Child ; China ; Female ; Humans ; Insulin ; metabolism ; Insulin Resistance ; Male ; Sensitivity and Specificity

OBJECTIVETo explore the role of epithelial sodium channel (ENaC) in regulating the functional activity of osteoclasts.

METHODSMultinucleated osteoclasts were obtained by inducing the differentiation of rat bone marrow cells with macrophage colony-stimulating factor (M-CSF) and RANKL. The osteoclasts were exposed to different concentrations of the ENaC inhibitor amiloride, and the expression of ENaC on osteoclasts was examined using immunofluorescence technique. The osteoclasts were identified with tartrate-resistant acid phosphatase (TRAP) staining, and the positive cells were incubated with fresh bovine femoral bone slices and the number of bone absorption pits was counted by computer-aided image processing. RT-PCR was performed to analyze the expression of cathepsin K in the osteoclasts.

RESULTSs Exposure to different concentrations of amiloride significantly inhibited the expression of ENaC and reduced the number of TRAP-positive osteoclasts. Exposure of the osteoclasts to amiloride also reduced the number of bone resorption pits on bone slices and the expression of osteoclast-specific gene cathepsin K.

CONCLUSIONs ENaC may participate in the regulation of osteoclast differentiation and bone resorption, suggesting its role in functional regulation of the osteoclasts and a possibly new signaling pathway related with ENaC regulation for modulating bone metabolism.

Animals ; Bone Marrow Cells ; cytology ; Bone Resorption ; Cathepsin K ; metabolism ; Cattle ; Cell Differentiation ; Epithelial Sodium Channels ; metabolism ; Macrophage Colony-Stimulating Factor ; metabolism ; Osteoclasts ; cytology ; RANK Ligand ; metabolism ; Rats ; Signal Transduction

This article reports that nano-silica solid dispersion technology was used to raise genistein efficiency through increasing the enzymatic hydrolysis rate. Firstly, genistin-nano-silica solid dispersion was prepared by solvent method. And differential scanning calorimetry (DSC) and transmission electron microscopy (TEM) were used to verify the formation of solid dispersion, then enzymatic hydrolysis of solid dispersion was done by snailase to get genistein. With the conversion of genistein as criteria, single factor experiments were used to study the different factors affecting enzymatic hydrolysis of genistin and its solid dispersion. And then, response surface method was used to optimize of nano-silica solid dispersion technology assistant enzymatic hydrolysis. The optimum condition to get genistein through enzymatic hydrolysis of genistin-nano-silica solid dispersion was pH 7.1, temperature 52.2 degrees C, enzyme concentration 5.0 mg x mL(-1) and reaction time 7 h. Under this condition, the conversion of genistein was (93.47 +/- 2.40)%. Comparing with that without forming the genistin-nano-silica solid dispersion, the conversion increased 2.62 fold. At the same time, the product of hydrolysis was purified to get pure genistein. The method of enzymatic hydrolysis of genistin-nano-silica solid dispersion by snailase to obtain genistein is simple, efficiency and suitable for the modern scale production.
Animals ; Calorimetry, Differential Scanning ; Genistein ; chemistry ; Hydrogen-Ion Concentration ; Hydrolysis ; Isoflavones ; chemistry ; Microscopy, Electron, Transmission ; Nanoparticles ; Phytoestrogens ; chemistry ; Silicon Dioxide ; chemistry ; Snails ; enzymology ; Solubility ; Technology, Pharmaceutical ; methods