[Correlation between accumulation of flavoneand expression of functional genes in Carthamus tinctorius].

The average yield of safflower blooming from 1 to 7 day was recorded and calculated, HPLC was used to detect the percentage composition of HYSA,quercetin,naringenin and kaempferol, and the real-time PCR was used to analyze the expression of chs and chi. The average yield,percentage composition of HYSA and naringenin as well as functional genes' expression presented similar trends. The average yield reached the highest peak at the third day, showing highpositive correlation with the contents of HYSA (r=0.756,P<0.05), and significant correlation with the expression of chi (r=0.892,P<0.01). The contents of naringenin showed a high positive correlation with the expression of chs(r=0.766,P<0.05). The study provides a theory basis for the composition and regulation mechanism of the flavonoid constituents and lays foundation for molecular mechanisms which lead to the difference of quality in C. tinctorius.

[Cloning,Bioinformatic Analysis of Chalcone-Flavonone Isomerase Gene( CHI) and Relationship Between Expression of CHI and Accumulation of HSYA in Carthamus tinctorius].

Objective: To clone chalcone-flavonone isomerase( CHI) gene in Carthamus tinctorius,to analyze the bioinformation of CHI,to compare the expression of CHI,and to analyze the percentage composition of hydroxysafflor yellow A( HSYA) during the florescence,in order to provide the foundation for functional verification of CHI and the composition and regulation mechanism of the flavonoid constituents in Carthamus tinctorius. Methods: CHI was cloned,bioinformatics was used to analyze the protein characteristics, real timePCR was used to analyze the expression of CHI,and HPLC was used to analyze the percentage composition of HSYA Results: A 696 bp CHI sequence in Carthamus tinctorius,the expression of CHI and the percentage composition of HSYA during the florescence were obtained. The variation tendency was similar between the expression of CHI and the accumulation of HSYA, which was increased gradually during 1 ~ 4 d and peaked at the fourth day,then decreased sharply during the fifth to the seventh day of florescence. Conclusion: This research provides a foundation for functional verification of CHI and the composition and regulation mechanism of the flavonoid constituents in Carthamus tinctorius.

Cyclic adenosine monophosphate differentiated beta-endorphin neurons promote immune function and prevent prostate cancer growth.

Pituitary adenylate cyclase-activating peptide (PACAP), a cAMP-activating agent, is highly expressed in the hypothalamus during the period when many neuroendocrine cells become differentiated from the neural stem cells (NSCs). Activation of the cAMP system in rat hypothalamic NSCs differentiated these cells into beta-endorphin (BEP)-producing neurons in culture. When these in vitro differentiated neurons were transplanted into the paraventricular nucleus (PVN) of the hypothalamus of an adult rat, they integrated well with the surrounding cells and produced BEP and its precursor gene product, proopiomelanocortin (POMC). Animals with BEP cell transplants demonstrated remarkable protection against carcinogen induction of prostate cancer. Unlike carcinogen-treated animals with control cell transplants, rats with BEP cell transplants showed rare development of glandular hyperplasia, prostatic intraepithelial neoplasia (PIN), or well differentiated adenocarcinoma with invasion after N-methyl-N-nitrosourea (MNU) and testosterone treatments. Rats with the BEP neuron transplants showed increased natural killer (NK) cell cytolytic function in the spleens and peripheral blood mononuclear cells (PBMCs), elevated levels of antiinflammatory cytokine IFN-gamma, and decreased levels of inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) in plasma. These results identified a critical role for cAMP in the differentiation of BEP neurons and revealed a previously undescribed role of these neurons in combating the growth and progression of neoplastic conditions like prostate cancer, possibly by increasing the innate immune function and reducing the inflammatory milieu.

Changes in thrombospondin-1 levels in the endothelial cells of the anterior pituitary during estrogen-induced prolactin-secreting pituitary tumors.

Thrombospondin-1 (TSP-1), a multifunctional matrix glyco-protein, has been shown to control tumor growth by inhibiting angiogenesis in various tissues. However, the role of this glycoprotein in pituitary angiogenesis is not well studied. In this report, we determined the changes in the production and action of TSP-1 on endothelial cells in anterior pituitary following estradiol treatment, which is known to increase prolactin-secreting tumor growth and vascularization in this tissue. We showed that TSP-1 immunoreactive protein is distributed in the anterior pituitary, particularly in the endothelial cells. Estradiol treatment for 2 and 4 weeks decreased the total tissue immunoreactive level of TSP-1 as well as the endothelial cell-specific immunoreactive level of this protein in the anterior pituitary. The steroid treatment also decreased the protein levels of TSP-1 in anterior pituitary tissues and in purified pituitary endothelial cells in primary cultures. Determination of the effects of TSP-1 on proliferation and migration of pituitary-derived endothelial cells in primary cultures elucidated an inhibitory action of TSP-1 on these vascular cell functions. These results suggest that locally produced TSP-1 may regulate estrogen angiogenic action on the pituitary.

Beta-endorphin modulation of interferon-gamma, perforin and granzyme B levels in splenic NK cells: effects of ethanol.

The effects of ethanol and beta-endorphin (beta-EP) on productions of cytolytic factors granzyme B, perforin and IFN-gamma in splenic rat NK cells were determined. Intracranial administration of beta-EP increased protein and mRNA levels of cytolytic factors in NK cells. Chronic ethanol feeding reduced the basal and beta-EP-induced levels of cytolytic factors in NK cells. In vitro treatment of beta-EP on NK cells increased the levels of perforin, granzyme B and IFN-gamma and their mRNA transcripts, whereas ethanol pre-treatment prevented beta-EP effects on cytolytic factors in these cells. These results suggest that beta-EP and ethanol interact to regulate NK cell functions.

Effects of sanguiin H-6, a component of Sanguisorbae Radix, on lipopolysaccharide-stimulated nitric oxide production.

The present study was conducted to evaluate the effect of sanguiin H-6, a component of Sanguisorbae Radix, on the production of nitric oxide (NO), using macrophages activated by lipopolysaccharide (LPS). Sanguiin H-6 inhibited nitrite production, taken as an index for NO, in a concentration-dependent fashion. This compound decreased inducible NO synthase (iNOS) activity, with the inhibitory effect at a concentration of 25 microM being equal to that of the known iNOS inhibitor aminoguanidine at 50 microM. However, unlike aminoguanidine, sanguiin H-6 was associated with improved cell viability. Reverse transcription-polymerase chain reaction analysis revealed that the expression of iNOS mRNA in activated macrophages was suppressed by sanguiin H-6 in a concentration-dependent manner. In addition, sanguiin H-6 even at a low concentration showed a clear scavenging effect on the NO generated from sodium nitroprusside (an NO donor). These findings indicate that not only does sanguiin H-6 act directly as an NO scavenger, but it also inhibits NO production in LPS-activated macrophages by the concomitant inhibition of iNOS mRNA induction and enzyme activity.

The RTEL1 rs6010620 polymorphism and glioma risk: a meta-analysis based on 12 case-control studies.

BACKGROUND: The association between the RTEL1 rs6010620 single nucleotide polymorphism (SNP) and glioma risk has been extensively studied. However, the results remain inconclusive. To further examine this association, we performed a meta-analysis. MATERIALS AND METHODS: A computerized search of the PubMed and Embase databases for publications regarding the RTEL1 rs6010620 polymorphism and glioma cancer risk was performed. Genotype data were analyzed in a meta-analysis. Odds ratios (ORs) with 95% confidence intervals (CIs) were estimated to assess the association. Sensitivity analyses, tests of heterogeneity, cumulative meta-analyses, and assessments of bias were performed in our meta-analysis. RESULTS: Our meta-analysis confirmed that risk with allele A is lower than with allele G for glioma. The A allele of rs6010620 in RTEL1 decreased the risk of developing glioma in the 12 case-control studies for all genetic models: the allele model (OR=0.752, 95%CI: 0.715-0.792), the dominant model (OR=0.729, 95%CI: 0.685-0.776), the recessive model (OR=0.647, 95%CI: 0.569-0.734), the homozygote comparison (OR=0.528, 95%CI: 0.456-0.612), and the heterozygote comparison (OR=0.761, 95%CI: 0.713-0.812). CONCLUSIONS: In all genetic models, the association between the RTEL1 rs6010620 polymorphism and glioma risk was significant. This meta-analysis suggests that the RTEL1 rs6010620 polymorphism may be a risk factor for glioma. Further functional studies evaluating this polymorphism and glioma risk are warranted.

CCDC26 gene polymorphism and glioblastoma risk in the Han Chinese population.

BACKGROUND: Glioblastoma (GBM) is an immunosuppressive tumor whose median survival time is only 12- 15 months, and patients with GBM have a uniformly poor prognosis. It is known that heredity contributes to formation of glioma, but there are few genetic studies concerning GBM. MATERIALS AND METHODS: We genotyped six tagging SNPs (tSNP) in Han Chinese GBM and control patients. We used Microsoft Excel and SPSS 16.0 statistical package for statistical analysis and SNP Stats to test for associations between certain tSNPs and risk of GBM in five different models. ORs and 95%CIs were calculated for unconditional logistic-regression analysis with adjustment for age and gender. The SHEsis software platform was applied for analysis of linkage disequilibrium, haplotype construction, and genetic associations at polymorphism loci. RESULTS: We found rs891835 in CCDC26 to be associated with GBM susceptibility at a level of p=0.009. The following genotypes of rs891835 were found to be associated with GBM risk in four different models of gene action: i) genotype GT (OR=2.26; 95%CI, 1.29-3.97; p=0.019) or GG (OR=1.33; 95%CI, 0.23-7.81; p=0.019) in the codominant model; ii) genotypes GT and GG (OR=2.18; 95%CI, 1.26-3.78; p=0.0061) in the dominant model; iii) GT (OR=2.24; 95%CI, 1.28-3.92; p=0.0053) in the overdominant model; iv) the allele G of rs891835 (OR=1.85; 95%CI, 1.14-3.00; p=0.015) in the additive model. In addition, "CG" and "CGGAG" were found by haplotype analysis to be associated with increased GBM risk. In contrast, genotype GG of CCDC26 rs6470745 was associated with decreased GBM risk (OR=0.34; 95%CI, 0.12-1.01; p=0.029) in the recessive model. CONCLUSIONS: Our results, combined with those from previous studies, suggest a potential genetic contribution of CCDC26 to GBM progression among Han Chinese.

In vitro and in vivo anti-diabetic activity of Swertia kouitchensis extract.

ETHNOPHARMACOLOGICAL RELEVANCE: Swertia kouitchensis has long been used as a folk medicine to treat hepatitis and diabetes in central-western China. Therefore, this study was aimed to evaluate the anti-diabetic activity of the plant ethanol extract. MATERIALS AND METHODS: Firstly, the extract was tested for its inhibitory activity on α-amylase and α-glucosidase in vitro. Following that, insulin secretion test in NIT-1 cell was performed. Then, oral sucrose or starch tolerance test of the extract were carried out in normal mice. After that, acute effect of the extract was executed in normal and streptozotocin-induced (60 mg/kg) diabetic mice. Eventually, long term effect of the extract was performed in diabetic mice for 4 weeks. Oral glucose tolerance test and biochemical parameters were estimated at the end of the study. RESULTS: Swertia kouitchensis extract could remarkably inhibit the activity of α-amylase and α-glucosidase and stimulate insulin secretion in vitro. And also the extract displayed anti-hyperglycemic activity, improved antioxidant capacity, ameliorated the hyperlipidemia and carbohydrate metabolism in diabetic mice. CONCLUSIONS: Swertia kouitchensis exhibits considerable anti-diabetic activity and metabolic alterations in diabetic mice. These results provide a rationale for the use of Swertia kouitchensis to treat diabetes mellitus.

Preventive effects of polysaccharides from Liriope spicata var. prolifera on diabetic nephropathy in rats.

This study was undertaken to investigate preventive effects of polysaccharides (LSP) from Liriope spicata var. prolifera on diabetic nephropathy in rats, which were induced by high fat-fed and low-dose streptozotocin (STZ). The levels of fasting blood glucose (FBG) and glycosylated hemoglobin (HbA1c) in diabetic rats were significantly decreased after treated with LSP for 28 days. Additional, the glucose tolerance of diabetes rats showed improvement after administration of LSP. The results also indicated that LSP were able to normalize hyperlipidemia, ameliorate oxidative stress, improve renal function parameters, inhibit the structural damages of kidney tissue and down-regulate the system of advanced glycation end products - receptor for advanced glycation end products (AGE-RAGE). In conclusion, LSP had potential preventive effects on diabetic nephropathy in diabetic rats.

Phylogenetic relationships among the colobine monkeys revisited: new insights from analyses of complete mt genomes and 44 nuclear non-coding markers.

BACKGROUND: Phylogenetic relationships among Asian and African colobine genera have been disputed and are not yet well established. In the present study, we revisit the contentious relationships within the Asian and African Colobinae by analyzing 44 nuclear non-coding genes (>23 kb) and mitochondrial (mt) genome sequences from 14 colobine and 4 non-colobine primates. PRINCIPAL FINDINGS: The combined nuclear gene and the mt genome as well as the combined nuclear and mt gene analyses yielded different phylogenetic relationships among colobine genera with the exception of a monophyletic 'odd-nosed' group consisting of Rhinopithecus, Pygathrix and Nasalis, and a monophyletic African group consisting of Colobus and Piliocolobus. The combined nuclear data analyses supported a sister-grouping between Semnopithecus and Trachypithecus, and between Presbytis and the odd-nosed monkey group, as well as a sister-taxon association of Pygathrix and Rhinopithecus within the odd-nosed monkey group. In contrast, mt genome data analyses revealed that Semnopithecus diverged earliest among the Asian colobines and that the odd-nosed monkey group is sister to a Presbytis and Trachypithecus clade, as well as a close association of Pygathrix with Nasalis. The relationships among these genera inferred from the analyses of combined nuclear and mt genes, however, varied with the tree-building methods used. Another remarkable finding of the present study is that all of our analyses rejected the recently proposed African colobine paraphyly and hybridization hypothesis and supported reciprocal monophyly of the African and Asian groups. SIGNIFICANCE: The phylogenetic utility of large-scale new non-coding genes was assessed using the Colobinae as a model, We found that these markers were useful for distinguishing nodes resulting from rapid radiation episodes such as the Asian colobine radiation. None of these markers here have previously been used for colobine phylogenetic reconstruction, increasing the spectrum of molecular markers available to mammalian systematics.

Coexistence of the antiferromagnetic and superconducting order and its effect on spin dynamics in electron-doped high-T(c) cuprates.

In the framework of the slave-boson approach to the t-t'-t''-J model, it is found that for electron-doped high- T(c) cuprates, the staggered antiferromagnetic (AF) order coexists with the superconducting (SC) order in a wide doping level ranged from underdoped to nearly optimally doped at the mean-field level. In the coexisting phase, it is revealed that the spin response is commensurate in a substantial frequency range below a crossover frequency ω(c) for all dopings considered, and it switches to the incommensurate structure when the frequency is higher than ω(c). This result is in agreement with the experimental measurements. Comparison of the spin response between the coexisting phase and the pure SC phase with a d(x(2)-y(2))-wave pairing plus a higher harmonics term (DP+HH) suggests that the inclusion of the two-band effect is important to consistently account for both the dispersion of the spin response and the non-monotonic gap behavior in the electron-doped cuprates.

Teratogenic potential in cultures optimized for oligodendrocyte development from mouse embryonic stem cells.

We describe a rapid and efficient 5-step program of defined factors for the genesis of brain myelin-forming oligodendrocytes (OLs) from embryonic stem cells (ESCs). The OLs emerge on the same time frame in vitro as seen in vivo. Factors promoting neural induction (retinoids, noggin) are required, while exogenous Sonic hedgehog is not. In contrast we were unable to generate OLs by trans-differentiation of ethically neutral mesenchymal stem cells, indicating a requirement for cis-differentiation via neural ectoderm for OL genesis. In the ESC-derived cultures, our optimized protocol generated a mixed population with 49% O4(+), Olig2(+) OL lineage cells. These cultures also retained pluripotential markers including Oct4, and an analysis of embryoid body formation in vitro, and allogeneic grafts in vivo, revealed that the ESC-derived cultures also retained teratogenic cells. The frequency of embryoid body formation from terminal differentiated OL cultures was 0.001%, 100-fold lower than that from ESCs. Our results provide the first quantitative measurement of teratogenicity in ESC-derived, exhaustively differentiated allogeneic grafts, and demonstrate the unequivocal need to purify ESC-derived cells in order to generate a safe population for regenerative therapy.

Spectral and optical properties in the antiphase stripe phase of the cuprate superconductors.

We investigate the superconducting order parameter, the spectral and optical properties in a stripe model with spin-(charge-) domain-derived scattering potential V(s) (V(c)). We show that the charge-domain-derived scattering is less effective than the spin scattering on the suppression of superconductivity. For [Formula: see text], the spectral weight concentrates on the (π,0) antinodal region and a finite energy peak appears in the optical conductivity with the disappearance of the Drude peak. But for V(s)≈V(c), the spectral weight concentrates on the (π/2,π/2) nodal region and a residual Drude peak exists in the optical conductivity without the finite energy peak. These results consistently account for the divergent observations in the ARPES and optical conductivity experiments in several high- T(c) cuprates and suggest that the 'insulating' and 'metallic' properties are intrinsic to the stripe state, depending on the relative strength of the spin- and charge-domain-derived scattering potentials.

[Cloning of ureI gene from Helicobacter pylori and its expression in E. coli].

OBJECTIVE: To clone the urea membrane channel gene (ureI) from Helicobacter pylori (Hp) for its expression in E. coli, and evaluate the expression conditions and immunological features of the fusion protein. METHODS: ureI gene cloned by PCR from Hp was inserted into the plasmid pET32a (+) to construct the recombinant plasmid pET32a/ureI, followed by identification by BglII and HindIII digestion and sequencing. E. coli BL-21+(DE3) was transformed with pET32a/ureI to obtain the engineered bacterium BL21+/UreI, which was cultured at different temperatures and induced with 1.0 mmol/L IPTG for expression of the recombinant protein. The expressed proteins were identified by SDS-PAGE and analyzed by Pro-gel analyzer 4.0. Western blotting was performed to evaluate the immunogenicity of the expressed protein. RESULTS: The cloned gene fragment was about 650 bp in length, and BglII and HindIII digestion of pET32a/ureI yielded a 650-bp band. Sequence analysis revealed that the cloned ureI gene contained 646 bp without reading frame alterations. Comparison against GenBank indicated a homology of 100% of the cloned gene with ureI gene of the corresponding Hp strains, and also one no less than 98.5% with ureI gene from other strains. The engineered E. coli BL21+/UreI could express recombinant UreI (rUreI) with His tag, and the target protein accounted for 20.2% of the total bacterial proteins after 1.0 mmol/L IPTG induction of the bacterium at 37 degrees C for 14 h. SDS-PAGE and Western blotting showed that the recombinant UreI protein was produced mainly in the inclusion bodies and fused with his-tag (rUreI/his), which could react with human anti-Hp and mAb to his tag but not with mAb to Hp UreB. CONCLUSIONS: We have successfully cloned ureI gene and constructed the prokaryotic expression plasmid for efficient rUreI expression, and the fusion protein rUreI/his expressed in the inclusion bodies can react specifically with both Hp antibody and his-tag antibody.

Ethanol suppression of the hypothalamic proopiomelanocortin level and the splenic NK cell cytolytic activity is associated with a reduction in the expression of proinflammatory cytokines but not anti-inflammatory cytokines in neuroendocrine and immune cells.

BACKGROUND: Immune signals activate a network of cytokines in the central nervous system (CNS) that in turn causes release of neurotransmitters and hormones to modulate immune cell functions. We have recently shown that hypothalamic beta-endorphin neurons, via inhibition of the sympathetic neuronal activity, activate natural killer (NK) cell function in the spleen, and this communication is disrupted following chronic ethanol administration. Beta-endorphin neuronal function is known to be regulated by various proinflammatory and anti-inflammatory cytokines. The effects of ethanol on the proinflammatory and anti-inflammatory cytokines known to control beta-endorphin neuronal and NK cell functions during immune challenges have not been determined. METHODS: In the present study, we evaluated the effects of chronic ethanol consumption on the basal and lipopolysaccharide (LPS)-activated NK cells' functions in the spleen, the beta-endorphin peptide precursor proopiomelanocortin (POMC) gene expression in the arcuate nucleus (ARC) of the hypothalamus, and mRNA levels of proinflammatory cytokines interleukin-1beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), and anti-inflammatory cytokines IL-6 and IL-10 in the spleen and in the ARC. Male rats were ad libitum fed rat chow (ad lib-fed), pair-fed an isocaloric liquid diet, or fed an ethanol-containing liquid diet, and each was treated with LPS (100 microg/kg body weight). After 2 hours, splenocytes and ARC tissues were isolated and used for this study. Splenocytes were used to determine mRNA levels of IL-1beta, TNF-alpha, IL-6, IL-10, granzyme B, and perforin using the real-time RT-PCR assays. Splenocytes were also used to determine the cytolytic activity using a standard 4-hour (51)Cr release assay against YAC-1 lymphoma target cells. Arcuate nuclei were used to determine IL-1beta, TNF-alpha, IL-6, IL-10, and POMC mRNA levels using real-time RT-PCR assays. RESULTS: The results demonstrate that ethanol feeding via a liquid diet for 2 weeks suppressed both basal and LPS-stimulated NK cell cytolytic functions and the levels of cytotoxicity-regulatory perforin and granzyme B mRNAs in the spleen. Ethanol feeding reduced the basal and LPS-stimulated levels of POMC mRNA in the ARC. Ethanol also impaired LPS-induced levels of IL-1beta and TNF-alpha mRNAs both in the spleen and in the ARC. In contrast, ethanol feeding did not cause any significant changes in basal and the LPS-stimulated expression of IL-6 and IL-10 mRNAs in the spleen and of IL-6 mRNA levels in the ARC. These results indicate that ethanol suppression of hypothalamic POMC levels and splenic NK cell functions is associated with a reduced expression of proinflammatory cytokines in neuroendocrine and immune cells.

Prenatal ethanol exposure alters the expression of period genes governing the circadian function of beta-endorphin neurons in the hypothalamus.

Sleep-wake disturbances and stress hyper-responsiveness have been observed in human neonates, children and adolescents who were exposed to alcohol during the prenatal period. Using the laboratory rat as an animal model, we investigated whether fetal ethanol exposure during gestational days 10-21 affects the circadian function of the stress-axis regulatory beta-endorphin neurons in the hypothalamus. Fetal ethanol-exposed rats showed abnormality in the circadian expression of proopiomelanocortin (POMC) mRNA encoding the peptide beta-endorphin in the arcuate nucleus of the hypothalamus during the adult period. These rats also showed altered circadian expression of the clock governing Period genes rPer1, rPer2 and rPer3, in the arcuate nucleus, and rPer1 and rPer 2 mRNA levels in the suprachiasmatic nucleus. Laser captured microdissection analysis identified constitutive expression of rPer1, rPer2 and rPer3 genes in beta-endorphin-containing neurons. These data suggest for the first time that fetal exposure to ethanol significantly alters the clock mechanisms governing the circadian function of beta-endorphin neurons.

Ethanol induces apoptotic death of developing beta-endorphin neurons via suppression of cyclic adenosine monophosphate production and activation of transforming growth factor-beta1-linked apoptotic signaling.

The mechanism by which ethanol induces beta-endorphin (beta-EP) neuronal death during the developmental period was determined using fetal rat hypothalamic cells in primary cultures. The addition of ethanol to hypothalamic cell cultures stimulated apoptotic cell death of beta-EP neurons by increasing caspase-3 activity. Ethanol lowered the levels of adenylyl cyclase (AC)7 mRNA, AC8 mRNA, and/or cAMP in hypothalamic cells, whereas a cAMP analog blocked the apoptotic action of ethanol on beta-EP neurons. The AC inhibitor dideoxyadenosine (DDA) increased cell apoptosis and reduced the number of beta-EP neurons, and it potentiated the apoptotic action of ethanol on these neurons. beta-EP neurons in hypothalamic cultures showed immunoreactivity to transforming growth factor-beta1 (TGF-beta1) protein. Ethanol and DDA increased TGF-beta1 production and/or release from hypothalamic cells. A cAMP analog blocked the activation by ethanol of TGF-beta1 in these cells. TGF-beta1 increased apoptosis of beta-EP neurons, but it did not potentiate the action of ethanol or DDA actions on these neurons. TGF-beta1 neutralizing antibody blocked the apoptotic action of ethanol on beta-EP neurons. Determination of TGF-beta1-controlled cell apoptosis regulatory gene levels in hypothalamic cell cultures and in isolated beta-EP neurons indicated that ethanol, TGF-beta1, and DDA similarly alter the expression of these genes in these cells. These data suggest that ethanol increases beta-EP neuronal death during the developmental period by cellular mechanisms involving, at least partly, the suppression of cAMP production and activation of TGF-beta1-linked apoptotic signaling.

[Expression and immunogenicity analysis of a recombinant fusion protein of V. Cholera ctB and H. pylori ure I].

AIM: To express fusion protein of the cholera toxin B subunit (ctB) and the urea membrane channel gene (ure I) of H. pylori in E. coli, and analyze its immunogenicity. METHODS: The prokaryotic expression vector pET32a+/ctB/ure I was constructed by inserting ctB gene amplified by PCR into the 5' terminus of ure I gene of expression vector pET32a+/ure I. The fusion gene was verified by endonuclease digestion and sequence analysis. The fusion protein ctB/ure I was expressed in E. coli BL21(DE3), purified by His-HP affinity chromatography, and analyzed by SDS-PAGE, Western blot and Pro-gel analyzer 4.0. The mice were immunized with purified ctB/ure I, and the immunoreactivity with ctB and ure I of the murine sera was analyzed by indirect ELISA. RESULTS: The pET32a+/ctB/ure I expression vector was constructed successfully and confirmed by endonuclease digestion and sequence analysis. The expressed ctB/ure I protein with molecular weight about 58,000 was shown when induced with 1 mmol/L IPTG for 4 h at 22 degrees C, and the protein could react with horse anti-ctB and human anti-ure I sera when detected with Western blot, and the purity of the purified protein was about 94.3%. The sera from mice immunized with purified ctB/ure I protein could react with ctB, ure I, and ctB/ure I when detected with indirect ELISA. CONCLUSION: The fusion protein expression vector pET32a+/ctB/ure I was constructed successfully. The fusion protein ctB/ure I was shown to have immunoreactivity with both anti-ctB and anti-ure I anti-sera, and could evoke production of anti-ctB and anti-ure I antibody in mice. Our work established a good foundation for further study on the new and effective H. pylori vaccines.

Protective effects of Acanthopanax Radix extract against endotoxemia induced by lipopolysaccharide.

Endotoxemia causes an enhanced production of reactive oxygen radicals, which contribute to multiple organ dysfunction. When rats were given intravenous lipopolysaccharide and tested 6 h later we found that the activities of catalase and glutathione peroxidase (GSH-Px) in kidney, were acutely suppressed while in serum the levels of nitric oxide (NO), lipid peroxidation, urea nitrogen and creatinine were significantly increased, indicating the excessive production of reactive oxygen radicals and the presence of renal injury. Pretreatment of rats with Acanthopanax Radix extract administered orally for 30 days reduced the NO and lipid peroxidation levels, increased the activities of catalase and GSH-Px, and attenuated the renal dysfunction. These results suggested that scavenging of reactive oxygen radicals is part of the mechanism by which Acanthopanax Radix extract works as an effective agent in preventing multiple organ dysfunction.