OBJECTIVEThe aim of the present study is to explore the effects of exhaustive exercise-induced oxidative stress on the antioxidant capacity and diformability of rat red blood cells.

METHODSRats were divided into three group (n = 10): sedentary control (C), exhaustive running exercise (ERE) and moderate running exercise (MRE) groups. Animals in the ERE group started treadmill running at a speed of 20 m/min speed with a 5% gradient, and reached a speed of 25 m/min with gradient 15% in 20 min. Running was continued until exhaustion. MRE group rats running at a speed of 20 m/min with a 5% gradient for 40 min. The levels of free thiol in erythrocyte membrane protein, lipidperoxidation levels and membrane protein components were analyzed. The red blood cell deformability of different groups was also observed.

RESULTSThe results showed that red blood cells were damaged by severe oxidative stress and the anti-oxidative capacity decreased significantly under exhaustive exercise conditions. Besides, lipid peroxidation and protein sulfhydryl cross-link based clustering of membrane were found after exhaustive exercise, and polymers high molecular weight (HMW) was formed. The elongation index (EI) was found to decline significantly in the ERE group compared with the C and MRE groups under shear stress (control group, 0.41 +/- 0.01 at 3 Pa and 0.571 +/- 0.008 at 30 Pa; ERE group, 0.314 +/- 0.013 at 3 Pa and 0.534 +/- 0.009 at 30 Pa; P < 0.05 and P < 0.01, respectively).

CONCLUSIONThese exercise-induced oxidative injure result in a significant decrease in deformability of rat erythrocytes, which in turn leads to dysfunction in the microcirculatory.

Animals ; Disease Models, Animal ; Erythrocyte Deformability ; Fatigue ; metabolism ; physiopathology ; Male ; Oxidative Stress ; Physical Conditioning, Animal ; Rats ; Rats, Sprague-Dawley

In order to affirm the cardioactive components in Fuzi, we identified a group of aminoalcohol- diterpenoid alkaloids in Fuzi using ultra high-performance liquid chromatography coupled with electrospray ionization mass spectrometer (UPLC-ESI-MS) method. Among a total of forty-one isolated ingredients, thirteen major aminoalcohol-diterpenoid alkaloids were identified by comparing their retention times and MS spectra with those of the reference substances. Moreover, Fuzi samples from different places of origin and with different processing methods were examined and their components displayed a pattern of high similarity, though the relative abundance varies probably due to their different processing methods. Furthermore, the cardiac effect of each identified alkaloid was individually evaluated using the isolated bullfrog heart perfusion experiment. Among the thirteen aminoalcohol diterpenoid alkaloids tested, six of them significantly enhanced the amplitude rates. Taken together, we affirm that the cardioactive components in Fuzi are aminoalcohol-diterpenoid alkaloids, shedding light on future studies of the mechanisms and development of these cardioactive compounds.
Aconitum ; chemistry ; Alkaloids ; chemistry ; Amino Alcohols ; chemistry ; Animals ; Cardiotonic Agents ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Heart ; drug effects ; In Vitro Techniques ; Plant Extracts ; chemistry ; Rana catesbeiana ; Spectrometry, Mass, Electrospray Ionization

BACKGROUNDIslet beta-cells are almost completely destroyed when patients with type 1 diabete are diagnosed. To date, insulin substitute therapy is still one of the main treatments. The cure of type 1 diabetes requires beta-cell regeneration from islet cell precursors and prevention of recurring autoimmunity. Therefore, beta-cell regeneration and proliferation emerge as a new research focus on therapy for type 1 diabetes. Islet beta-cell regeneration and development are controlled by many growth factors, especially insulin-like growth factor-1 (IGF-1).

METHODSRecombinant adenovirus encoding rat IGF-1 (rIGF-1) was constructed and transduced into rat beta-cells, RINm5F cells. Western blotting analysis and ELISA were used to detect rIGF-1 protein. Streptozotocin (STZ) was used to induce RINm5F cell destruction. The level of nitric oxide (NO) was detected in cell culture supernatants by the Griess reaction. Islet cell function was evaluated by glucose-stimulated insulin production. Flow cytometry analysis was further used to investigate the apoptosis of RINm5F cells. Thiaoollyl blue viability assay was applied to determine cell viability.

RESULTSThe recombined adenovirus-rIGF-1 was successfully constructed and the titer was 4.0 x 10(8) pfu/ml. The rIGF-1 protein was effectively expressed in the RINm5F cells and cell culture supernatants. rIGF-1 expression remarkably inhibited STZ-induced islet cell apoptosis and significantly decreased the level of NO. Furthermore, IGF-1 expression also significantly protected insulin secretion and cell proliferation in a time-dependent manner.

CONCLUSIONSOur study suggests that locally produced rIGF-I from RINm5F cells may be beneficial in maintaining beta-cell function, protecting beta-cells from the destruction of apoptosis factors and promoting beta-cell survival and proliferation. IGF-I might be considered as a candidate gene in gene therapy for type 1 diabetes. In addition, it appears that the apoptosis induced by STZ may be NO-dependent.

Adenoviridae ; genetics ; Animals ; Antibiotics, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Cell Line ; Cell Proliferation ; Cell Survival ; Flow Cytometry ; Humans ; Insulin-Like Growth Factor I ; genetics ; physiology ; Insulin-Secreting Cells ; cytology ; drug effects ; metabolism ; Rats ; Reverse Transcriptase Polymerase Chain Reaction ; Streptozocin ; pharmacology

OBJECTIVETo construct the recombinant adenovirus containing rat insulin-like growth factor 1 (rIGF-1), and then infect to rat islet beta cells-RINm5F cells with the virus to investigate the role of rIGF-1 to streptozotocin-induced cell impairment in vitro.

METHODSRecombinant adenovirus encoding rIGF-1 was constructed, and then infect to RINm5F cells. rIGF-1 protein was detected by Western blot analysis and ELISA method. Then streptozotocin was used to induce RINm5F cells impairment. The levels of nitric oxide were detected in cells culture supernatants. The cells function was evaluated by glucose-stimulated insulin production. The apoptosis was analyzed by flow cytometry. Thiaoollyl blue viability assay was applied to exam the number of viable cells.

RESULTSThe recombined adenovirus-rIGF-1 was constructed successfully and its titer was about 4.0x10(8) pfu/ml. The rIGF-1 was expressed in the RINm5F cells and cell culture supernatants. rIGF-1 expression could inhibit islet cells apoptosis, significantly decrease the level of NO induced by streptozotocin and significantly increase insulin secretion and cells viability.

CONCLUSIONThese results suggested that the high concentration of rIGF-1 in cultured islets may be beneficial in maintaining islet beta cells function and protecting islet beta cells from apoptosis-mediated factors. The apoptosis induced by STZ may be NO-dependent.

Adenoviridae ; genetics ; metabolism ; Animals ; Apoptosis ; drug effects ; Cell Line ; Cell Survival ; drug effects ; Diabetes Mellitus ; genetics ; therapy ; Gene Expression ; Gene Transfer Techniques ; Genetic Therapy ; Genetic Vectors ; genetics ; metabolism ; Humans ; Insulin ; metabolism ; Insulin-Like Growth Factor I ; genetics ; metabolism ; pharmacology ; Insulin-Secreting Cells ; cytology ; drug effects ; metabolism ; Rats ; Streptozocin ; pharmacology

The aim of this study is to investigate the effect of monoammonium glycyrrhizinate (MAG) on lipopolysaccharide (LPS) -induced acute lung injury (ALI) and its anti-inflammatory mechanism in mice. All male ICR mice were randomly divided into six groups: LPS group; control group; MAG 3, 10, and 30 mg x kg(-1) groups; and dexamethasone (DXM) 5 mg x kg(-1) group. Lung dry weight and wet weight percentage and permeability were detected. Neutrophil infiltration in bronchoalveolar lavage fluid (BALF) and lung tissues was detected by cell count and morphological analysis. The levels of TNF-alpha and IL-10 in lung were detected by ELISA. MPO activity was determined followed the specification. MAG induced a decrease in lung wet weight/dry weight ratio, and significantly decreased in total leucocyte number and neutrophil percentage in the BALF, and MPO activity of lung in a dose-dependent manner. Importantly, It could up-regulate the IL-10 level and down-regulate the TNF-alpha level in the lung tissue of ALI mice. These results suggested that the protective effect of MAG in mice on LPS induced ALI was associated with the regulation of TNF-alpha/IL-10 balance, and MAG maybe a potentially treatment for ALI/ARDS.
Acute Lung Injury ; chemically induced ; metabolism ; pathology ; Animals ; Anti-Inflammatory Agents ; pharmacology ; Bronchoalveolar Lavage Fluid ; cytology ; Glycyrrhizic Acid ; pharmacology ; Interleukin-10 ; metabolism ; Leukocyte Count ; Lipopolysaccharides ; Lung ; pathology ; Male ; Mice ; Mice, Inbred ICR ; Neutrophils ; pathology ; Organ Size ; drug effects ; Peroxidase ; metabolism ; Protective Agents ; pharmacology ; Tumor Necrosis Factor-alpha ; metabolism

Child, Preschool ; Diagnosis, Differential ; Female ; Humans ; Rickets ; diagnosis ; etiology ; therapy ; Treatment Outcome ; Vitamin D Deficiency ; complications

OBJECTIVETo investigate the association between Fc gamma receptor IIA gene polymorphism and susceptibility to chronic periodontitis in Chinese Han nationality.

METHODSDNA samples were collected with buccal swabs from 63 patients with severe chronic periodontitis(CP), 103 patients with mild to moderate CP and 80 healthy individuals as control. Polymorphism in Fc gamma receptor IIA gene cluster was analyzed with PCR-SSP. The genotype distribution and allele frequency among different groups were compared.

RESULTSIt was found that the frequency of Fc gamma RIIA-R/R131 genotype was significantly higher in patients with severe CP (19.05%) compared to that of the healthy controls (P < 0.0125).

CONCLUSIONThe Fc gamma RIIA-R/R131 genotype may be one of the contributors for the increased susceptibility to severe CP in Chinese Han nationality.

Adult ; Antigens, CD ; genetics ; China ; ethnology ; Chronic Disease ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Male ; Middle Aged ; Periodontitis ; genetics ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic ; Receptors, IgG ; genetics

Brain Neoplasms ; Germinoma ; Humans ; Pineal Gland

OBJECTIVETo develop a tight tetracycline-controlled HCV-C double transgenic mouse model.

METHODSBy crossbreeding of ApoE-rtTA-tTS transgenic mice with TRE-HCV-C transgenic mice, the double transgenic mice were produced in the F1 generation. The presence of HCV-C and tTS gene in the F1 generation was confirmed by PCR, followed by further identification and quantification of the transgene using Southern blot hybridization. The expression of HCV-C in the liver of the mouse model was detected immunohistochemically.

RESULTS AND CONCLUSIONTwo transgenic mice were obtained, which contained ApoE-rtTA-tTS and TRE-HCV-C genes in the genome. Five founders contained HCV-C gene as confirmed by PCR and Southern blot hybridization. The tight tetracycline-controlled system may facilitate further study of HCV-C gene expression and gene therapy of hepatic cellular carcinoma.

Animals ; Apolipoproteins E ; genetics ; Blotting, Southern ; Breeding ; Crosses, Genetic ; Female ; Gene Expression Regulation, Viral ; drug effects ; Hepacivirus ; genetics ; immunology ; Hepatitis C Antigens ; genetics ; immunology ; Male ; Mice ; Mice, Transgenic ; Polymerase Chain Reaction ; Tetracycline ; pharmacology ; Trans-Activators ; genetics ; Viral Core Proteins ; genetics

A detection method for furazolidone and florfenicol in soil with various environmental matrices was established using ultra performance liquid chromatography-tandem spectrometry (UPLC / MS / MS) technique. Extracting solution of a mixture of phosphate buffer (pH = 3) and acetonitrile (3 : 7, V/ V) was used in this experiment. The extracted water samples were enriched by SAX-HLB solid phase extraction column before the process of nitrogen blowing ( high purity nitrogen). The enriched antibiotics were desalted with 8 mL of methanol. Waters BEH C18(2. 1 × 100 mm) column was used for the sample separation. UPLC / MS / MS was carried out for qualitative and quantitative analysis under multi-reaction monitoring mode. The detection limit of the method was determined by 3 times of signal-to-noise ratio, and the limit of determination of the method was determined by 10 times of signal-to-noise ratio. The results showed that the detection limits of furazolidone and florfenicol were 1. 19 and 0. 41 μg / kg, respectively, and the limits of quantitation of furazolidone and florfenicol were 3. 40 and 1. 37 μg / kg, respectively. Besides, recovery experiment showed that, for the soil samples spiked with 50 μg / L furazolidone and florfenicol, the recoveries were 79% for florfenicol and 92% for furazolidone. Similarly, for the soil samples spiked with 200 μg / L furazolidone and florfenicol, the recoveries were 96% for furazolidone and 86% for florfenicol.