Objective:To observe the hypoglycemic effects of Cortex Lycii Radicis (CLR) on alloxan induced diabetic mice.Methods:The alloxan induced diabetic mice were given drug decoction of CLR[2.5g/(kg?d) -1 , or 5.0g/(kg?d) -1 )] for successive 2 weeks, The blood sugar level was determined with GOD method.Results:There were remarkedly differences in the levels of blood sugar between before and after treatment of various groups. ( P

Objective To study the protective effects of taurine on liver and kidney injury induced by intestinal ischemia-reperfusion(I/R) in rats.Methods Male Wistar rats were randomly assigned into Sham,I/R,and taurine groups.Thirty min before operation,2% taurine(200 mg/kg) was injected via dorsal vein of the rat′s penis.Intestinal ischemia-reperfusion was produced by occlusion of superior mesenteric artery for one hour later,then the blood flow was restored by removing the clamps.Blood samples were taken from rats in I/R and taurine groups at 1.5,3,6 and 12h after reperfusion,and the serum levels of ALT,AST,BUN and Cr were measured to evaluate the functions of liver and kidney.Tissues from livers and kidneys were cryostated and stained with hematoxylin and eosin to observe changes in histological pathology.TUNEL was also performed to examine apoptotic cells and the average light density levels were measured.Results The serum levels of ALT,AST,BUN and Cr in I/R group were significantly higher than those in Sham group(P

Objective To construct the recombinant eukaryotic expression vector pRNAT-U6,1- siEdg4 which curries small interfering RNA(siRNA)of Edg4 and observe the silencing effect of Edg4 gene targeted siRNA in ovarian cancer cell line SKOV3.Methods The Edg4 gene-targeted hairpin siRNA sequence was designed according to the Edg4 sequence in Genbank,and the two complementary oligo nucleotide strands were synthesized and annealed and inserted into the pRNAT-U6.1 plasmid to build a recombinant Edg4 siRNA eukaryotic expression vector,which was sequenced and identified to contain the correct Edg4 siRNA sequence.The human ovarian carcinoma cell lines SKOV3 were transfeeted with the vector using lipofeetamine method.The efficiency of transfecting cells was observed with fluorescent microscope and the mRNA expression level of Edg4 gene was detected by real time quantitative PCR.The LPA levels in cell supernatants were detected using a biochemical method.And the apoptosis of SKOV3 cells induced by the vector was evaluated by flow cytometry.Results The recombinant eukaryotic expression vector was confirmed to contain correct Edg4 siRNA sequence by PCR and sequencing.After transfection large amounts of green fluorescence were seen in plasma and nuclei of SKOV3 cells and the positive cell rates were 64%.The expression level of Edg4 mRNA in transfeeted SKOV3 cell line was significantly decreased (0.05?0.01 vs 0.29?0.04,P

Objective It is rarely reported whether myricetin inhibits the activation and function of cardiac fibroblasts and thereby prevents myocardial fibrosis. This study was to investigate the effects of myricetin on the activation,proliferation and secretion of cardiac fibroblasts and its possible molecular mechanisms. Methods Fibroblasts isolated from 1-3 days old rats were cultured and their activation,proliferation and secretion were induced with the transforming growth factor (TGF). The fibroblasts were incubated with myricetin at different concentrations of 1,3,10,30 and 100 μmol/L for 24 hours followed by detection of their proliferation with the CKK8 kit,the transcription levels of fibrotic factors by RT-PCR and the expression levels of α-SMA and signal proteins by immunoflu-orescence staining and Western blot,respectively. Results The expression of α-SMA was significantly up-regulated in the cardiac fi- broblasts of the rats in the TGF-β,30 μmol/L myricetin+TGF-β and 100 μmol/L myricetin+TGF-β groups as compared with that in the control group (P<0.05) but down-regulated in the 30 μmol/L myricetin+TGF-β and 100 μmol/L myricetin+TGF-β groups in com-parison with that in the TGF-β group (P<0.05). At 48 hours,the transcription levels of collagenⅠ,collagenⅢ,fibronectin and con-nective tissue growth factor were markedly higher in the TGF-β,30 μmol/L myricetin+TGF-β and 100 μmol/L myricetin+TGF-β groups than in the control group (P<0.05) but lower in the 30 μmol/L myricetin+TGF-β and 100 μmol/L myricetin+TGF-β groups than in the TGF-β group (P<0.05). The phosphorylation levels of smad2 and smad3 were remarkably elevated in the TGF-β,30 μmol/L myricetin+TGF-β and 100 μmol/L myricetin+TGF-β groups and the expression of smad4 reduced in the TGF-β group as com-pared with the control group (P<0.05). The levels of smad2,smad3 and smad4 were all significantly decreased in the 30 μmol/L myr-icetin+TGF-β and the 100 μmol/L myricetin+TGF-β groups in comparison with the TGF-β group (P<0.05). Conclusion Myricetin suppresses the activation,proliferation and secretion of cardiac fibroblasts induced by TGF-β via inhibiting the smad signaling pathway.

BACKGROUNDSepticemia and inflammation-mediated septic shock caused by Vibrio vulnificus (VV) is strongly associated with chronic liver disease. This study examined the effects of antimicrobial therapy on expression of hepatic toll-like receptors and inflammatory cytokines in rats with alcohol-induced liver disease complicated by VV sepsis.

METHODSMale Sprague-Dawley rats were assigned to the following treatment groups: normal control (N), alcoholic liver disease control (A), antimicrobial-treated alcoholic liver disease control (AA), alcoholic liver disease with VV sepsis (AV), and antimicrobial-treated alcoholic liver disease with VV sepsis (AVA). Alcohol-induced liver disease was observed in all groups except N. Expression of mRNAs encoding hepatic toll-like receptors 2 and 4, myeloid differentiation protein-2, tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, IL-6 and IL-10 was determined by RT-PCR.

RESULTSmRNAs encoding toll-like receptors 2 and 4 and myeloid differentiation protein-2 were significantly up-regulated in group AV as compared to control groups at 2 - 24 hours of sepsis; peak expression occurred at 12 hours. These mRNAs were also up-regulated in group AVA but to lesser degrees than in group AV at comparable time post-infection. mRNAs encoding TNF-alpha, IL-1beta and IL-6 were significantly elevated in group AV as a function of infection. In group AVA as compared to AV, expression of TNF-alpha and IL-1beta mRNAs was lower at 12 - 24 hours post-infection and expression of IL-6 mRNA was lower at 24 hours post-infection. Compared with control groups, IL-10 mRNA expression in group AV was markedly higher at 12 - 24 hours of sepsis. Expression of IL-10 mRNA was lower in group AVA as compared to AV at 24 hours of sepsis.

CONCLUSIONSAntimicrobial therapy reduces expression of toll-like receptors and cytokines in rats with alcohol-induced liver disease complicated by VV sepsis. Monitoring hepatic toll-like receptor and cytokine expression during antibiotic therapy may be valuable for determining the course of VV sepsis in subjects with liver disease.

Adaptor Proteins, Signal Transducing ; genetics ; Animals ; Anti-Infective Agents ; therapeutic use ; Cytokines ; genetics ; Interleukin-10 ; genetics ; Interleukin-1beta ; genetics ; Interleukin-6 ; genetics ; Liver ; drug effects ; metabolism ; Liver Diseases, Alcoholic ; drug therapy ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Sepsis ; drug therapy ; genetics ; microbiology ; Toll-Like Receptor 2 ; genetics ; Toll-Like Receptor 4 ; genetics ; Toll-Like Receptors ; genetics ; Tumor Necrosis Factor-alpha ; genetics ; Vibrio Infections ; drug therapy ; Vibrio vulnificus ; physiology

OBJECTIVETo investigate the dynamic changes of oxidative stress and nuclear factor-E2 related factor 2 (Nrf2) expression in the lung tissues of acute hydrogen sulfide (H2S) intoxicated rats and intervention effects of ulinastatin (UTI).

METHODSA total of 96 SD rats of clean grade were divided randomly into four groups: normal control group (n = 8), UTI control group (n = 8), H2S -intoxicated model group (n = 40), and UTI treatment group (n = 40). The H2S-intoxicated model group and UTI treatment group were exposed to H2S (283.515 mg/m3) by inhalation for 1h, then UTI treatment group was intraperitoneally exposed to UTI at the dose of 10(5) U/kg for 2 h. H2S-intoxicated model group and UTI treatment group were sacrificed at 2, 6, 12, 24 and 48 h after exposure, respectively. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and glutathione (GSH) in the rat lung tissues were measured. The expression levels of Nrf2 mRNA in the rat lung tissues were detected. Pathological changes of rat lung tissues were observed under a light microscope and the lung injury scores were evaluated.

RESULTSCompared with control group, the pulmonary SOD, CAT and GSH levels at 2,6 and 12 h after exposure and the pulmonary GSH-Px levels at 2, 6, 12 and 24 h after exposure in H2S-intoxicated model group significantly decreased (P < 0.05 or P < 0.01). The levels of pulmonary MDA at 2, 6, 12 and 24 h after exposure in H2S-intoxicated model group were significantly higher than those in normal control group (P < 0.01). As compared with H2S -intoxicated model group, the pulmonary GSH-Px activities at 6 and 12 h after exposure, the pulmonary CAT activities at 2, 6 and 12 h after exposure, the pulmonary GSH levels at 2, 6, 12 and 24 h after exposure and the pulmonary SOD activities at 2, 6, 12, 24 and 48 h after exposure in UTI treatment group significantly increased (P < 0.05 or P < 0.01), the pulmonary MDA levels at 2, 6 and 12 h after exposure in UTI treatment group significantly decreased (P < 0.01). The expression levels of Nrf2 mRNA at 2, 6, 12, 24 h after exposure in H2S-intoxicated model group were 0.314 +/- 0.011, 0.269 +/- 0.010, 0.246 +/- 0.011 and 0.221 +/- 0.018, respectively, which were significantly higher than those (0.149 +/- 0.012) in control group (P < 0.01). As compared with H2S-intoxicated model group, the expression levels (0.383 +/- 0.017, 0.377 +/- 0.014, 0.425 +/- 0.017, 0.407 +/- 0.011 and 0.381 +/- 0.010) of Nrf2 mRNA at 2, 6, 12, 24 and 48 h after exposure in UTI treatment group significantly increased (P < 0.01). The lung injury at 24 h after exposure in H2S-intoxicated model group was higher than that in UTI treatment group. Histopathological examination showed that the scores of lung injury at 12, 24 and 48 h after exposure in UTI treatment group was significantly lower than those in H2S-intoxicated model group (P < 0.01).

CONCLUSIONOxidative stress and Nrf2 activation may be the important factors in rat lung injury induced by H2S-intoxicated, UTI may reduce the rat lung injury and protect the rat lung from damage induced by H2S by inhibiting ROS, improving the imbalance in redox and up-regulating Nrf2 mRNA expression.

Acute Lung Injury ; chemically induced ; metabolism ; Animals ; Glycoproteins ; pharmacology ; Hydrogen Sulfide ; poisoning ; Lung ; metabolism ; Male ; NF-E2-Related Factor 2 ; metabolism ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo detect the effects of antimicrobial agents on the toll-like receptor (TLR) and so on in liver tissue of rats after intragastric infusion with alcohol with vibrio vulnificus (VV) sepsis.

METHODSSprague-Dawley rats were randomly divided into normal control group (N group, n = 6), rats after intragastric infusion with alcohol control group (group A, n = 6), drug intervention on rats after intragastric infusion with alcohol control group (group AA, n = 6), rats after intragastric infusion with alcohol with VV sepsis group (group AV, n = 24, killed at 2, 6, 12, 24 hours after injecting VV respectively, six rats per group), as well as drug intervention on rats after intragastric infusion with alcohol with vibrio vulnificus sepsis group (group AVA, n = 30, killed at 6, 12, 24 hours and one week after injecting VV respectively, six rats per group). The expressions and dynamic changes of TLR4 mRNA and so on by RT-PCR in liver tissue of each group were measured.

RESULTSThe expressions of TLR4 mRNA in AV-6 hours group was 0.775 +/- 0.101, the expressions of TLR4 mRNA in AVA-6 hours group was 0.600 +/- 0.064; the expressions of TLR4 mRNA in AV-12 hours group was 0.918 +/- 0.133, the expressions of TLR4 mRNA in AVA-12 hours group was 0.583 +/- 0.112; the expressions of TLR4 mRNA in AV-24 hours group was 0.732 +/- 0.110, the expressions of TLR4 mRNA in AVA-24 hours group was 0.512 +/- 0.118. Compared with AV group, the expressions of TLR4 mRNA in liver diminished greatly in AVA group at 6, 12 and 24 hours after being injected with VV (AVA-6 hours group compare with AV-6 hours group, t = -3.573, P < 0.01; AVA-12 hours group compared with AV-12 hours group, t = - 4.722, P < 0.01; AVA-24 hours group compare with AV-24 hours group, t = - 3.340, P < 0.01).

CONCLUSIONThe treatment with antibacterial agents may reduced the expression of TLR and so on in liver of rats after intragastric infusion with alcohol with VV sepsis. The treatment with antibacterial agents may regulate the balance of the inflammatory response in VV sepsis and generate the visible therapeutical effect for VV sepsis.

Animals ; Disease Models, Animal ; Ethanol ; Interleukin-10 ; metabolism ; Liver ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Sepsis ; immunology ; metabolism ; Toll-Like Receptors ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Vibrio vulnificus

OBJECTIVETo explore the expression diversification of CD4(+)CD25(+)CD127(low) regulatory T (Treg) cells and Foxp3 mRNA in the peripheral blood of children with aplastic anemia after the treatment with cyclosporine.

METHODSFifty children with chronic aplastic anemia were enrolled, among whom 30 received cyclosporine treatment (cyclosporine group) and 20 were treated with conventional methods (conventional group). Twenty healthy children were enrolled as the control group. The expression of CD4(+)CD25(+)CD127(low) Treg cells was detected by flow cytometry. The expression of Foxp3 mRNA was detected by real-time Q-PCR.

RESULTSThe expressions of Foxp3 mRNA and CD4(+)CD25(+)CD127(low)Treg cells showed no significant difference between the cyclosporine and the control groups 6 months after treatment. On the contrary, there were significantly lower expressions of both in the conventional group than in the control group (P<0.05). Meanwhile, the cyclosporine group had significantly higher expressions of Foxp3 mRNA and CD4(+)CD25(+)CD127(low) Treg cells than the conventional group (P<0.05).

CONCLUSIONSThe expressions of CD4(+)CD25(+)CD127(low) Treg cells and Foxp3 mRNA in children with aplastic anemia increase after cyclosporine treatment.

Adolescent ; Anemia, Aplastic ; drug therapy ; immunology ; Child ; Child, Preschool ; Chronic Disease ; Cyclosporine ; pharmacology ; therapeutic use ; Female ; Forkhead Transcription Factors ; blood ; genetics ; Humans ; Immunosuppressive Agents ; pharmacology ; Male ; RNA, Messenger ; blood ; T-Lymphocytes, Regulatory ; drug effects

OBJECTIVETo examine the time trends of perinatal mortality and the frequency of birth defects occurring in perinatal deaths, and to provide a national perspective on the impacts of congenital anomalies on perinatal mortality from 1990 through 2001.

METHODSData were from Chinese Birth Defects Monitoring network-a hospital-based congenital anomalies registry system. During 1990 - 2001, all live or still births with 28 weeks of gestation or more, born in monitoring units, were studied within 7 days after delivery. The proportion of perinatal deaths due to birth defects, which was defined as the number of perinatal deaths associated with congenital anomalies per 100 perinatal deaths, was calculated by birth area (urban versus rural), geographic-economic status (coast areas, inner land areas and remote areas), to evaluate the impacts of birth defects on perinatal mortality.

RESULTSPerinatal mortality declined from 22.85 per 1000 in 1990 to 13.26 per 1000 in 2001, which showed a significant downward trend. Similar trend was also observed in the rate of stillbirth and the ratio of early neonatal death. However, the proportion of perinatal deaths due to birth defects had an increasing trend although the perinatal birth defects-specific death rate was declining, especially during 1996 - 2001. This result was also seen in urban and rural area, in coast regions, in inner land regions and in remote regions of China. Higher rate of birth defects occurring in perinatal deaths was observed in urban area than in rural area. Significant difference of this rate was also found among different geographic-economic regions, with the highest one in inland regions.

CONCLUSIONBirth defects were accounted for an increasing proportion of perinatal deaths in China, and had become one of the major causes of perinatal deaths.

China ; epidemiology ; Congenital Abnormalities ; classification ; epidemiology ; Female ; Humans ; Infant Mortality ; trends ; Infant, Newborn ; Pregnancy ; Risk Factors ; Time Factors

OBJECTIVETo determine the mutations pattern of the genes of a collodion baby.

METHODSCollodion baby is a genetic heterogeneous disease caused by mutations of several genes. Since the most common mutations were observed in TGM1 gene, this gene was chosen for mutation screening. The screening was carried out by PCR and direct sequencing. The allele specific primers were designed for a missense mutation and allele-specific (AS) PCR was carried out in 50 normal individuals for population study.

RESULTSThree novel alterations were detected in TGM1 gene of the proband, a missense mutation c.463C > T (p.Arg155Trp) in exon 3, a nonsense mutation c.578G > A (p.Trp193X) in exon 4, and a single nucleotide deletion (c.694delG) also in exon 4 of TGM1 gene. This infant's father was heterozygote of c.694delG mutation, while his mother carried the two mutations (c.463C > T and c.578G > A) on the same chromosome. The missense mutation was not detected in his father and in any of the control individuals by AS-PCR.

CONCLUSIONThree novel mutations were identified in TGM1 gene in a Chinese collodion baby. A double mutation (c.463C > T and c.578G > A) located on the maternal allele while the c.694delG deletion on the paternal allele.

Alleles ; DNA Mutational Analysis ; Exons ; Genes, Recessive ; Genetic Testing ; Humans ; Ichthyosis, Lamellar ; genetics ; Male ; Point Mutation ; Polymerase Chain Reaction ; Sequence Analysis ; Sequence Deletion