Adolescent ; Bacteria ; isolation & purification ; Child ; Child, Preschool ; Humans ; Infant ; Infant, Newborn ; Respiratory Tract Infections ; microbiology

OBJECTIVETo investigate the correlation of the changes in CD8(+)CD28(-) T cell percentage with platelet (PLT) and D-dimer (D-D) levels in patients with multiple injuries (MI).

METHODSTwenty-six patients with MI, 31 with a single injury (SI group) and 26 healthy individuals were examined for peripheral blood CD8(+)CD28(-) T cells and intracellular transformation growth factor-β1 (TGF-β1) and interleukin 10 (IL-10) contents using flow cytometry at 24, 48, and 72 h after the injuries. PLT and D-dimer levels were compared among the 3 groups.

RESULTSCD8(+)CD28(-) T cells, TGF-β1 and IL-10 were significantly higher in MI group than in SI group and healthy control group (P<0.05) without significant differences between the latter 2 groups. The levels of PLT and D-D differed significantly among the 3 groups, the highest in MI group and the lowest in the control group. In MI group, CD8(+)CD28(-) T cells, TGF-β1 and IL-10 significantly increased at 48 h after the injury (P<0.05) but decreased significantly at 72 h (P<0.05) compared with the measurements at 24 h. The levels of PLT and D-D trended to decrease with time after the injuries and showed significant differences among the 3 groups at any of the 3 time points (P<0.05). CD8(+)CD28(-) T cells, TGF-β1 and IL-10 were all positively correlated with the levels of PLT and D-D in MI patients (r>0.70, P<0.05 for all comparisons).

CONCLUSIONIn MI patients, CD8(+)CD28(-) T cell percentage and their cytokines tend to increase early after the injury but decrease significantly at 72 h in close relation with the changes of the coagulation function following the injuries.

CD28 Antigens ; metabolism ; CD8 Antigens ; metabolism ; Case-Control Studies ; Fibrin Fibrinogen Degradation Products ; metabolism ; Flow Cytometry ; Humans ; Interleukin-10 ; metabolism ; Multiple Trauma ; immunology ; T-Lymphocyte Subsets ; cytology ; Transforming Growth Factor beta1 ; metabolism

AIMTo discuss the effect of Pitavastatin on angiogenesis in vivo and its mechanism in Klotho heterozygous deficient mice.

METHODSThe heterozygous deficient Klotho mice (kl +/-) and wild mice (kl +/+) from the same litter were used to establish the animal model of hind-limb ischemia and grouped into control and Pitavastatin group, respectively. Hind-limb blood flow was evaluated using Laser Doppler perfusion imager (LDPI) before treatment and after operation of hind-limbs. The capillaries in muscle of limbs were counted by means of CD-31 labeled immuno-fluorescence. The phosphorylation of Akt (Protein kinase B) in cells was measured by direct immunohistochemical technique. The expression of vascular endothelial growth factors (VEGFs) in muscle of limbs was assessed using Western blotting.

RESULTSAfter treatment of Pitavastatin, the blood flow in ischemic limbs of the Kl +/- and wild mice improved obviously, the ratio of blood flow area in ischemic limb to that in non-ischemic limb increased and the density of capillaries increased in ischemic limbs of the Kl +/- and wild mice. Pitavastatin enhanced the phosphorylation of Akt and the expression of VEGF in ischemic limbs of the Kl +/- and wild mice.

CONCLUSIONPitavastatin has the pro-angiogenesis effect in vivo and the VEGF-p-Akt-NO pathway may be involved in the mechanism of the effect of Pitavastatin.

Angiogenesis Inducing Agents ; pharmacology ; Animals ; Heterozygote ; Ischemia ; Male ; Mice ; Mice, Knockout ; Quinolines ; pharmacology ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVEWe investigated the effects of pitavastatin on angiogenesis and perfusion in C3H/He mice with unilateral hind limb ischemia.

METHODSC3H/He mice treated with saline (n = 15) or pitavastatin (1 mg.kg(-1).d(-1), n = 15) per gavage for 1 week underwent unilateral hind limb ischemia surgery and were treated for another 5 weeks. Hind-limb blood flow was measured by Laser Doppler perfusion imager (LDPI, ischemic/nonischemic limb, %) at baseline, immediately after ischemia and weekly thereafter for 5 weeks. Endpoints included local vessel counts by immunofluorescence, phospho-Akt positive cell counts by immunoenzyme histochemical technique, vascular endothelial growth factors (VEGFs) expression in ischemic limbs by Western blot and serum nitric oxide metabolite (NOx) by chrome dioxide Griess method.

RESULTSLower extremity perfusion was significantly improved in pitavastatin treated mice vs. controls as measured by LDPI% at 1 week post ischemia and thereafter (P < 0.05). Pitavastatin treatment was associated with significantly increased capillary count [(47 +/- 11) vs. (26 +/- 14)/per high-power field (x 200), P < 0.05] and greater percentage of phospho-Akt positive cells [(6 +/- 1) vs. (2 +/- 0)/per high-power field (x 200), P < 0.05] in ischemic limbs. Serum NOx [(77.3 +/- 21.8) vs. (52.1 +/- 11.2) mol/L, P < 0.05) and VEGF protein expression in ischemic limbs were also significantly increased in pitavastatin group than those in control group.

CONCLUSIONSPitavastatin enhances angiogenesis and perfusion in CsH/He mice with limb ischemia.

Animals ; Disease Models, Animal ; Ischemia ; physiopathology ; Lower Extremity ; blood supply ; Male ; Mice ; Mice, Inbred C3H ; Neovascularization, Physiologic ; drug effects ; Nitric Oxide ; blood ; Quinolines ; pharmacology ; Vascular Endothelial Growth Factors ; metabolism

Objective To explore the relationship between plasticity ofhippocampus neuronal morphology and pathogenesis of epilepsy by observing the changes of mossy fiber sprouting (MFS) and P-glycoprotein (P-gp) expression in the hippoeampus of rat models of amygdala-kindling epilepsy.Methods Ninety male Wistar rats were randomly divided into epilepsy model group (n=40),drug treatment group (n=40) and sham-operated group (n=10).Models of chronic epilepsy were established by stimulating the amygdale; rats in the drug treatment group were perfused antiepileptic drug levetiracetam into stomach [100 mg/(kg·d),twice daily].At different observation time points (1,2,4 and 8 weeks after the treatment),Timm staining was employed to observe the changes of MFS; immunohistochemical method was used to detect the dynamic changes of P-gp.Results (1) After the success of model making,abnormal MFS levels in the hippocampal CA3 transparent layer were noted; lowest MFS scores were noted in S1 subgroup,which showed no significant difference as compared with those in sham-operated group (P＞0.05); the MFS scores in S2,S4 and S8 subgroups increased gradually with a peak level at 8 weeks after inducement,which showed significant difference as compared with those in sham-operated group (P＜0.05).No significant differences on MFS grading scores were noted in the drug treatment group between each two time points (P＞0.05).(2) The P-gp expression in the epilepsy model group showed significant difference at different time points (P＜0.05); highest expression level was noted at one week after the surgery,and returned to normal at four weeks.However,the P-gp expression showed no significant difference in the drug treatment group between each two time points (P＞0.05),except for 1 week after treatment.Conclusion MFS is the important mechanism of chronic epilepsy,and P-gp is the product of epilepsy,which is the main reason of epilepsy drug resistance.

OBJECTIVEThrough maternal inheritance, to explore the genetic structures and relationships of Dong, Gelao, Tujia and Yi ethnic population in Guizhou of China.

METHODSThe mtDNA D-loop hypervariable segment I (HVS I ) in 108 samples of four ethnic populations were sequenced. Then, the nucleotide diversity was estimated and a phylogenetic tree was constructed by Neighbor-Joining method.

RESULTSIn the detected 497 bp fragments, 86 polymorphic sites were found, and 82 different haplotypes were identified. The phylogenetic tree of four ethnic populations showed: Yi, Tujia and Gelao clustered more closely than Dong did.

CONCLUSIONYi and Tujia population are very closely related, the reason may be that they either originate from a common ancestry or frequently undergo the gene exchanges and admixtures. The genetic relationship between Tujia and Gelao population is nearer, perhaps because they have settled in the adjacent regions. Dong and Yi population show the farthest genetic relationship, this is probably due to their different historical origins and geographic segregation.

Base Sequence ; China ; DNA, Mitochondrial ; chemistry ; classification ; genetics ; Ethnic Groups ; genetics ; Genetic Variation ; Humans ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid

OBJECTIVETo investigate the mechanism by which heat shock protein 90 (HSP90) regulates 26S proteasome in hyperthermia.

METHODSHyperthermic HepG2 cell models established by exposure of the cells to 42 degrees celsius; for 3, 6, 12, and 24 h were examined for production of reactive oxygen species (ROS) and cell proliferation, and the changes in Hsp90α and 26S proteasome were analyzed.

RESULTSROS production in the cells increased significantly after hyperthermia (F=28.958, P<0.001), and the cell proliferation was suppressed progressively as the heat exposure time extended (F=621.704, P<0.001). Hyperthermia up-regulated Hsp90α but decreased the expression level (F=164.174, P<0.001) and activity (F=133.043, P<0.001) of 26S proteasome. The cells transfected with a small interfering RNA targeting Hsp90α also showed significantly decreased expression of 26S proteasome (F=180.231, P<0.001).

CONCLUSIONThe intracellular ROS production increases as the hyperthermia time extends. Heat stress and ROS together cause protein denature, leading to increased HSP90 consumption and further to HSP90 deficiency for maintaining 26S proteasome assembly and stability. The accumulation of denatured protein causes unfolded protein reaction in the cells to eventually result in cell death.

HSP90 Heat-Shock Proteins ; metabolism ; Hep G2 Cells ; Hot Temperature ; Humans ; Proteasome Endopeptidase Complex ; metabolism ; RNA, Small Interfering ; genetics ; Reactive Oxygen Species ; metabolism ; Up-Regulation

OBJECTIVETo study how hepatitis B virus(HBV) 'a' determinant hotpoint mutations were influecing the hepatitis B vaccine efficacy.

METHODSPrimers were designed in HBV conservative region, and the degenerate probes for detecting 16 'a' determinant hotpoint mutations were developed for gene chips. Sensitivity and specificity of the gene chips were evaluated by clone sequencing. Sera of 47 pairs of mothers and infants with immune failure and 323 mothers of children with immune protection of HB vaccine were detected by the gene chips.

RESULTSResult from clone sequencing demonstrated that the gene chips were specific for the detection of 'a' determinant hotpoint mutations. The wild type of HBV was still dominant, with the prevalence of 78.66%, and the mutation frequencies of 126A, 145R, 126S-1, 126S-2, 129H, 144A, and 129R were 11.27%, 5.76%, 5.28%, 4.56%, 1.20%, 0.72% and 0.24%, respectively. The prevalence of 126A mutation was significantly higher than that of other mutations(P < 0.01). No significant differences were found in mother-infant transmission rates of 126A, 126S-1, 126S-2 and 145R variants.

CONCLUSIONThe currently available hepatitis B vaccine could block mother-infant transmission of 126A, 126S and 145R variants. It appears that there is no need to develop a new hepatitis B vaccine against 126 and 145 variants at present, but the consistent epidemiological surveillance on HBV mutants should be carried out.

Adult ; Female ; Genotype ; Hepatitis B ; prevention & control ; transmission ; Hepatitis B Vaccines ; immunology ; Hepatitis B virus ; genetics ; immunology ; Humans ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; prevention & control ; Mutation ; Oligonucleotide Array Sequence Analysis ; Pregnancy ; Pregnancy Complications, Infectious ; prevention & control ; virology

AIM To study the chemical constituents from the heartwood of Dalbergia cochinchinensis Pierre ex Laness.METHODS The 70%ethanol extract from the heartwood of D.cochinchinensis was isolated and purified by silica gel,Sephadex LH-20 and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULT Twenty-three compounds were isolated and identified as 3-O-acetylbetulin aldehyde(1),2,2'-oxybis(1,4-di-tert-butylbenzene)(2),ethyl 4-hydroxybenzoate(3),1-acetyl-β-carboline(4),7-hydroxydihydroflavone(5),palmic acid(6),hexadeca-4,7-diene(7),linoleic acid(8),methyl 4-hydroxybenzoate(9),2-(2-hydroxy-1-methyl-2-phenythyl)-4,5-dimethoxyphenol(10),2-methoxy-3-hydroxyxanthone(11),dibutyl terephthalate(12),6,4'-dihydroxy-7-methoxyflavan(13),pteroyanin G(14),benzoic acid,4-ethoxy-2-methoxy-,methyl ester(15),liquiritigenin(16),4,2',5'-trihydroxy-4'-methoxychalcone(17),7-hydroxy-6-methoxyflavone(18),6,4'-dihydroxy-7-methoxyflavone(19),2'-hydroxyformonetin(20),3'-methoxyformonetin(21),3'-hydroxyformonetin(22),6,7,4'-trihydroxyflavanone(23).CONCLUSION Compounds 2,4 are isolated from genus Dalbergia for the first time.Compounds 6-8,19,21 are isolated from this plant for the first time.

This study was purposed to explore the mechanisms of cladribine (2-CdA)-inducing apoptosis of multiple mycloma RPMI 8226 cells. The MTT method was used to determine cell proliferation after being treated with 2-CdA. Apoptosis and cell cycle progression were examined by flow cytometry. Transmission electron microscopy was used to observe ultrastructural changes of RPMI 8226 cells. RT-PCR and Western blot were used to analyze the mRNA and protein expression levels of BCL-2, MCL-2 and caspase-3 respectively. The results showed that the 2-CdA inhibited proliferation of RPMI 8226 cells in time and dose-dependent manner. Typical apoptotic morphological and ultrastructure changes could be observed by electron microscopy. Flow cytometry showed that 2-CdA induced myeloma cell apoptosis and arrested myeloma cells in the G2/M phase. The mRNA expression of BCL-2 and MCL-1 decreased but that of caspase-3 not apparently changed. Western blot results suggested that the change trend of BCL-2 MCL-1 and caspase-3 was the same as result of RT-PCR. It is concluded that 2-CdA exhibits inhibitory effects on RPMI 8226 cells in vitro. Activating the mitochondrial and death receptor pathways of apoptosia may be the potential mechanism, meanwhile, the cell cycle arrest may also play a critical role in apoptosis.
Apoptosis ; drug effects ; Caspase 3 ; Cell Division ; Cell Line, Tumor ; Cell Proliferation ; Cladribine ; pharmacology ; Humans ; Multiple Myeloma ; pathology ; Proto-Oncogene Proteins c-bcl-2