Objective To explore the relationship between the expression of nestin protein and the expression of vascular endothelia growth factor(VEGF)in hypoxia-ischemia brain damage(HIBD)of neonatal SD rats.Methods Fifty-six 7-day postnatal SD rats were randomly divided into 2 groups,sham operation(n=28)and hypoxic ischemic(HI)group(n=28).Animals were killed in 0,3,12,24 h,3,7,14 d after HIBD rat models established.The brain tissue was obtained to make paraffin section.Expressions of nestin and VEGF protein were examined with immunohistochemical staining and image quantitative analysis.Results The number of VEGF positive cell in the hippocampus and cortex increased reactively after HI and reached peak at the 12th hour and persistent to the 24th hour,which returned the normal level on d3,however,the number of nestin positive cell reached peak on d7 and returned the normal level on d14 after HI.Conclusions VEGF and nestin may contribute to neurolproctetive effects against HIBD.Studying the way to facilitate the coexpression of the nestin and VEGF may have important rolls to enhance the repairment and regeneration.

Objective To investigate the correlation between the incidence and characteristics of anemia in chronic heart failure population through retrospective analysis of patients hospitalized at the Hunan Provincial People's Hospital.Methods A total of 259 patients who were diagnosed as chronic heart failure (CHF) and hospitalized at the Hunan Provincial People's Hospital from October to November in 2012 was collected,and divided into cardiac function Ⅰ ～ Ⅳ grade,then their clinical characteristics was analyzed.Results (1)Compared with CHF patients without anemia,the creatinine value was significantly higher,the percentage of patients with severe heart failure was significantly higher in the CHF patients with anemia (P＜ 0.05).(2) Logistic regression analysis demonstrated that age (95％ CI:1.123 ～ 3.580,P ＜ 0.05),renal insufficiency (95％ CI:1.320 ～ 4.845,P ＜ 0.05),and cardiac function (95 ％ CI:1.368 ～ 3.385,P ＜ 0.05) in patients with heart failure were important risk factors for anemia.Conclusions Anemia was common in CHF patients.The poorer cardiac function was and the lower hemoglobin was,the higher incidence of anemia was.The patients with anemia had the characteristics of worse renal function and poorer nutritional status compared patients without anemia.Age,renal insufficiency,and cardiac function were important risk factors for anemia.

DEK protein's carboxy-terminal DNA-binding region(CBD)is a newly found DNA-binding domain of DEK,which contains several phosphorylation sites and has a close correlation with DEK protein's function in vivo and in vitro.Using prokaryotic expression system,the peptide of DEK protein's carboxy-terminal DNA-binding region(CDB)was expressed and purified.In detail,the CDB DNA fragment was constructed into pET-30a(+)vector,and E.coli BL21(DE3)competent cells were used as host cells.The fusion protein His-CBD was expressed by induction of IPTG and purified by Ni-NTA agarose.The result of SDS-PAGE showed that the molecular weight of the purified protein was about 10.7kDa.Electrophoretic mobility shift assay(EMSA)indicated that DEK-CDB prefered to bind to supercoiled form of DNA in vitro,it had similar character to the binding of whole length DEK protein with DNA.This suggested that the carboxy-terminal DNA-binding region of DEK protein might function on the binding of DEK protein to DNA partly.

Objective:To investigate the effect of miR-154 on the apoptosis of hepatic stellate cells (HSC-T6). Methods:Hepat-ic stellate cells (HSC-T6) were transfected with miR-154 mimics and miR-154 inhibitor, and the cells were trandfected with mimics NC and inhibitor NC as the negative control. The effects of miR-154 on the proliferation of HSC-T6 were detected at different time points by CCK-8. A flow cytometry with double staining of Annexin and PI was used to detect the cell cycle and apoptosis rate of HSC-T6. Results:The proliferation ability of the cells was increased,the apoptosis rate was decreased significantly, and the proportion of cells in G0/G1 phase were decreased, and those in G2/M phase were increased significantly after transfected with miR-154 mimics. The proliferation ability of the cells was decreased,the apoptosis rate was increased significantly, and the proportion of cells in G0/G1 phase were increased, and those in G2/M phase were decreased significantly after transfected with miR-154 inhibitor. Conclusion:MiR-154 can promote the proliferation of HSC-T6 and inhibit the apoptosis of HSC-T6.

Objective:To summarize the single nucleotide polymorphisms ( SNPs) in ABCC2 and the effect on clinical drug appli-cation. Methods:According to the related articles published in domestic and abroad, the correlation between the single nucleotide pol-ymorphisms in ABCC2 and drug responses was classified and summarized. Results:ABCC2 translocator played an important role in the transmembrane transport of many physiological compounds and exogenous compounds. Numerous studies have demonstrated that the single nucleotide polymorphisms in ABCC2 possibly affected the expression or activity of ABCC2, which leading to the variation in the absorption, distribution and excretion of certain drugs and toxicants. However, the limitation and controversy were still emerged in the results. Conclusion:The influence of ABCC2 single nucleotide polymorphisms on clinical drug application shows significantly referen-tial value for the guidance of medication and the evaluation of efficacy, however, it can not be used as the only indicator yet.

Objective To study the relationship between tyrosine phosphorylation(TP)and protein expression of insulin receptor substrate-1(IRS-1)and insulin resistance in patients with gestational diabetes mellitus(GDM).Methods IRS-1 expression and TP in skeleton muscle tissue were determined by Western blot and immunoprecipitation in women with GDM(GDM group,n=22),normal pregnant women(normal pregnancy group,n=22)and normal nonpregnant women(normal nonpregnant group,n=13).Fasting plasma glucose(FPG)and fasting insulin(FINS)were measured by oxidase assay and immunoradioassay. Results(1)The levels of FPG,FINS,and insulin resistance index were calculated according to homeostasis model assessment [ HOMA-IR;(5.6?0.8)mmol/L,(15.4?5.1)mU/L,and 1.2?0.5 ] in GDM group were significantly higher than those in normal pregnancy group [(4.4+0.5)mmol/L,(10.6 ?3.1)mU/L,and 0.8?0.3;P

A new caffeate compound, (E)-erythro-syringylglyceryl caffeate (1), was isolated from the roots and rhizomes of Nardostachys chinensis Batal., together with nine known phenolic compounds, including (+)-licarin A (2), naringenin 4', 7-dimethyl ether (3), pinoresinol-4-O-β-D-glucoside (4), caraphenol A (5), Z-miyabenol C (6), protocatechuic acid (7), caffeic acid (8), gallic acid (9) and vanillic acid (10). Their chemical structures were elucidated on the basis of spectroscopic data and physicochemical properties. Furthermore, this is the first report of compounds 2, 5 and 6 from Nardostachys genus.
Caffeic Acids ; chemistry ; isolation & purification ; Flavanones ; chemistry ; isolation & purification ; Furans ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Hydroxybenzoates ; chemistry ; isolation & purification ; Lignans ; chemistry ; isolation & purification ; Nardostachys ; chemistry ; Plant Roots ; chemistry ; Rhizome ; chemistry ; Vanillic Acid ; chemistry ; isolation & purification

Objective To study the content of volatile oil from old leaves, tender leaves, fallen leaves and seeds of cinnamomum camphora tree in Hunan, China. To prepare and identify the volatile oil of its β-cycoldextrin inclusion compound. Methods The volatile oil was extracted by water-steam distillation. With inclusion rate as the index, trituration method, saturated water solution method and ultrasound method were compared. The optimum conditions were investigated by the orthogonal test. The inclusion compound was identified by thin-layer chromatography (TLC), ultraviolet visible spectrum (UV-Vis) and differential scanning calorimetry (DSC). Results From June to July, the average content of volatile oil extracted from old leaves, tender leaves, fallen leaves and seeds were 1.58%, 1.52%, 0.84% and 1.39%, respectively. The optimum preparation conditions were as follows:the ratio of β-cyclodextrin and volatile oil was 10∶1, the adding water was 4 times and inclusing time was 2 h. Before and after inclusion, the spectrum of TLC and UV-Vis of volatile oil showed no obvious change. The DSC of inclusion compound,β-cycoldextrin, volatile oil and the mixture had significant differences. Conclusion The amount of the volatile oil from old leaves is higher. The optimized condition of inclusion is stable and reasonable.

SUMMARY Objective:To investigate the influence of cryoprotectants and cooling rates in vitrificationmethod on the spindles of rabbit M Ⅱ oocytes.Methods:Rabbit oocytes were verified by using cryoloopwith ethylene glycol(EG)singly or EG combined with dimethyl sulphoxide(DMSO)as cryoprotectants,and cooled by taking oocytes directly into liquid nitrogen or by vitrification machine.After frozen rabbit o-ocytes thawed,the microtubulin and chromosome of the spindles were fixation and stained by immunofluo-rescent method.Confocal microscope was used to reveal spindle configuration.Results:In the two proto-cols of single EG used and EG combined with DMSO,the spindles were severely injured.But in protocolof EG combined with DMSO and at ultra-rapid cooling rate,the normal configuration of spindle rate ofthawed rabbit oobytes was similar to that of the control group.Conclusion:The protocol of EG combinedwith DMSO as cryoprotectants and with extremely high cooling rate by vitrification machine can producethe best effect on conservation of spindle configuration in vitrification of rabbit oocytes.

Objective To investigate the effects of CD4+T cell depletion in BALB/c mice on the immunogenicity and virulence of replication-competent recombinant vaccinia virus. Methods Twelve BALB/c mice were inoculated with recombinant Tiantan vaccinia ( rTV, n=8 ) or vaccinia virus Tiantan strain ( VTT, n=4) by tail scarification after the depletion of CD4+T cells with anti-CD4 monoclonal anti-body( McAb) injected intraperitoneally for three days before virus inoculation.A control group without anti-body treatment was set up accordingly.Several parameters including the body weight, pocks on tail, viral shedding and the percentage of CD4+T cells were monitored.In the fourth week after virus infection, ovaries were collected from mice and viral loads in the tissue were titrated by plaque forming assay on chick embryo fibroblast ( CEF) cells.The specific cellular immune responses against vaccinia virus and HIV induced by inoculation of VTT and rTV respectively were detected by intracellular cytokine staining ( ICS) assay.ELISA was used to detect antibodies against vaccinia virus and HIV-1 gp120.Results All mice with or without CD4 McAb treatment showed typical poxvirus pocks on tails after inoculation of vaccinia viruses, but none of them developed secondary or satellite lesions.It took a longer time for CD4+T cell-depleted mice to heal from lesions, to regain body weights and to release viruses than the mice in control group.No vaccinia virus was detected in the ovaries of CD4+T cell-depleted mice or mice in control group.The mean absorbance( A) values for the detection of HIV-specific and vaccinia virus-specific antibodies in CD4+T cell-depleted mice with ELISA were respectively 0.119 and 0.168, which were significantly lower than those in mice of control group.The titers of neutralizing antibodies against vaccinia virus in McAb/rTV treated mice (1 ∶321) were lower than those in rTV treated mice (1 ∶1286) (P<0.05).The percentages of CD4+T cells secreting IFN-γ(0.654%) in McAb/rTV treated mice were significantly lower than those in rTV treated mice in the fourth week after immunization (P <0.0004).No significant differences with the vaccinia virus-specificCD8+ T cell responses were observed among mice with or without CD4+T cells depletion.Conclusion Thereplication and dissemination of replication-competent recombinant vaccinia virus could be effectively controlledin the mice with CD4+ T cell-depletion.The depletion of CD4+ T cells significantly reduced the humoraland CD4+ T cell responses, but had no effect on CD8+ T cell responses.