Transplanted neuronal precursors migrate and differentiate in the developing mouse brain

The subventricular zone (SVZ), lining the lateral ventricle in forebrain, retains a population of neuronalprecursors with the ability of proliferation in adult mammals. To test the potential of neuronal precursorsin adult mice, we transplanted adult SVZ cells labeled with fluorescent dye PKH26 into the lateral ventricleof the mouse brain in different development stages. The preliminary results indicated that the graftedcells were able to survive and migrate into multiple regions of the recipient brain, including SVZ, the thirdventricle, thalamus, superior colliculus, inferior colliculus, cerebellum and olfactory bulb etc; and the amountof survival cells in different brain regions was correlated with the development stage of the recipient brain.Immunohistochemical studies showed that most of the grafted cells migrating into the specific target couldexpress neuronal or astrocytic marker. Our results revealed that the neuronal precursors in adult SVZstill retained immortality and ability of proliferation, which is likely to be induced by some environmentalfactors.

Influence of Moxibustion Serum on the Expression of Fas bcl-2 mRNA and Protein of EL-4 Lymphoma Cells

Objective： To observe the influence of moxibustion serum of mice on the expression of Fas, bcl-2 mRNA and protein of EL-4 lymphoma cells in vitro. Method： The EL-4 lymphoma cells were cultivated for 24 h in the moxibustion serum of mice. The expression of Fas and bcl-2 mRNA of EL-4 lymphoma cells were detected by in-situ hybridization method, and the protein expression of Fas and bcl-2 were observed by the immuocytochemistry method. Results： The expression of bcl-2 mRNA and protein decreased , and the expression of Fas mRNA and protein increased significantly in EL-4 cells, which were cultivated in the moxibustion serum compared those cultivated in normal mice serum （P〈0.05）. Conclusion： Moxibustion serum could down-regulate the bcl-2 mRNA and protein and up-regulate the Fas mRNA and protein of EL-4 cells.

Effect of quercetin on the expression of Bcl-2/Bax apoptotic proteins in endometrial cells of lipopolysaccharide-induced-abortion mice

OBJECTIVE: To explore the effect of quercetin on the expressions of Bcl-2/Bax apoptotic proteins in endometrial cells in mice with abortion induced by lipopolysaccharide.METHODS: For in vivo experiment, twenty five Kunming mice were randomly divided into five groups at day 4 of pregnancy, with 5 mice per group. The mice were treated with lipopolysaccharide(LPS)through tail vein intravenous injection at day 4 of pregnancy, followed by different concentrations of quercetin by oral gavage consecutively at days 5 to6 of pregnancy. On day 7 of gestation, the mice were sacrificed and the histopathological changes of the uterus tissues were observed. Immunohistochemical staining was applied to the detection of Bcl-2/Bax apoptotic proteins in the endometrial cells. For in vitro experiment, the primary endometrial cells werecultured using a uterus tissue mass culturing method sampled at day 4.5 of pregnancy. The cells were treated with LPS with or without different dosages of quercetin, respectively, for 12 h after 80% confluence. The expression of Bcl-2/Bax apoptotic proteins were detected by western blotting.RESULTS: Both the in vivo and in vitro experiments showed decreased expression of Bcl-2 and enhanced expression of Bax after LPS treatment, leading to a decreased Bcl-2/Bax ratio. The expression of Bcl-2 significantly increased while the expression of Bax was significantly elevated, in the LPS plus quercetin group compared to the LPS only group.CONCLUSION: These results suggest that quercetin has protective effect by partially regulating the expression of Bcl-2/Bax proteins, which in turn inhibits endometrial cell apoptosis and benefits the embryo implantation.

Anti-tumor effect of the extract from Qingyihuaji formula on pancreatic cancer by down-regulating Notch-4 and Jagged-1

OBJECTIVE： To investigate, in terms of Notch signaling pathway, the effect on pancreatic cancer of the extract of an anti-tumor prescription -- Qingyihuaji formula （QYHJ） -- from Traditional Chinese Medicine （TCM）.METHODS： Nude mice were implanted subcutaneously with human pancreatic cancer cell line SW1990 and then randomly divided into four groups： Control, QYHJ extract, Gemcitabine, and Combination of QYHJ extract and gemcitabine. Treatments were given for 21 days and tumor growth was evaluated simultaneously. Then, expression of Notch receptors （Notch-I, Notch-2, Notch-3, and Notch-4） and their Jagged ligands （Jagged-1 and Jagged-2） in dissected tumor tissue were detected by real-time quantitative reverse transcription-polymerase chain reaction and Western blot. Finally, immunohistochemistry was performed to detect CD133, a marker of pancreatic cancer stem cells （CSCs）, to evaluate the impact of QYHJ extract on pancreatic CSCs.RESULTS： QYHJ extract treatment effectively inhib- ited the tumor growth in nude mice. The expression of both Notch-4 and Jagged-1 were decreased significantly in QYHJ treatment groups （P 〈 0.05）, while gemcitabine alone had no significant effect in down-regulating Jagged-1 （P 〉 0.05）. No significant difference was observed in the ex- pression of Notch-1, Notch-2, Notch-3, and Jagged-2 between three treatment groups and control group （P 〉 0.05）. Moreover, immunohistochemical analysis showed that the number of CD133 positive cells was significantly reduced by QYHJ treatment （P 〈 0.05）, and the combined treatment was more effective than gemcitabine alone （P 〈 0.05）.CONCLUSION： The role of the extract in pancreatic cancer treatment was associated with down-regulation of Notch-4 and Jagged-1 in Notch signaling pathway. The extract could enhance the antitumor activity of gemcitabine and was more effective than gemcitabine in regulating Notch signaling pathway to some extent.

Daifan San intervenes in forkhead box P3 and the interleukin(IL)-23/IL-17A signaling pathway to help prevent and treat primary biliary cirrhosis

OBJECTIVE:To investigate the mechanism by which Daifan San(DFS)prevents and treats primary biliary cirrhosis(PBC)via the forkhead box P3(FoxP3)and interleukin(IL)-23/IL-17A signaling pathways.METHODS:Ninety C57BL/6 mice were randomly divided into the control,model,DFS low-dose,DFS middle-dose,DFS high-dose and ursodeoxycholic acid(UDCA)groups(n=15 per group).A mouse model of PBC was induced using polyinosinic polycytidylic acids(poly I:C).Lymphocyte subset expression in the peripheral blood was analyzed via flow cytometry.The inflammatory cytokines and antimitochondrial autoantibody(AMA)levels were detected via enzyme-linked immunosorbent assays.The expressions and location of typeⅠcollagen,typeⅢcollagen,cytokeratin 19 and FoxP3 in the liver tissue were evaluated via immunohistochemistry.FoxP3,IL-23 and IL-17 expressions in the peripheral blood and liver tissue were evaluated via real-time polymerase chain reaction and western blotting.RESULTS:IL-17,IL-23,IL-8,IL-33,TNF-α,and AMA expressions were significantly increased in the model group and decreased in the DFS and UDCA groups.Conversely,Treg cell and FoxP3 expressions were significantly decreased in the model group and increased in the DFS and UDCA groups.The IL-23/IL-17A signaling pathway was closely correlated with chronic inflammation of the bile duct in PBC and functional deletion of Treg cells,leading to reduced FoxP3 levels and mediating the loss of tolerance in PBC.CONCLUSION:DFS may delay the occurrence and relieve the symptoms of PBC by downregulating IL-23/IL-17A signaling pathway expression and upregulating FoxP3 expression.

Effect of Gubenfangxiao decoction on respiratory syncytial virus-induced asthma and expression of asthma susceptibility gene orosomucoid 1-like protein 3 in mice

OBJECTIVE： To investigate the effect of Gubenfangxiao decoction （GBFXD） on respiratory-syncytial-virus （RSV） -induced asthma and the expression of asthma susceptibility gene, orosomucoid 1-1ike protein 3 （ORMDL3） in mice. METHODS： Seventy-two female BALB/c mice were randomly assigned to normal, model, GBFXD high dose, GBFXD moderate dose, GBFXD low dose and montelukast groups. An asthma model was induced via intraperitoneal injection and aerosol inhalation of ovalbumin （OVA） and repeated intranasal instillation of RSV in all mice, except those in the normal group. All treatments were administered at the first onset of asthma （within 8 weeks of model establishment） and the mice were euthanized after 28 days of treatment. The levels of transforming growth factor-β （TGF-β） and interleukin-6 （IL-6） in bronchoalveolar lavage fluid （BALF） of the mice were measured and the expression of asthma sus- ceptibility gene ORMDL3 in lung tissue was deter- mined using real-time polymerase chain reaction （RT-PCR） and western blotting. RESULTS： Expression of ORMDL3 and levels of TGF-β and IL-6 were significantly higher in the mod- el group （P 〈 0.05, P 〈 0.01） compared with the normal mice. Levels of ORMDL3, TGF-β and IL-6 were significantly lower in all three GBFXD treated groups （P 〈 0.05） compared with the model group. However, the levels in the GBFXD treatment groups did not differ significantly from the montelukast group. CONCLUSION： GBFXD had a therapeutic effect in this experimental model. The functional mechanism of GBFXD may involve multiple factors, including alleviation of airway inflammation, down-regulation of asthma susceptibility gene ORMDL3 and inhibition of airway remodeling.

Effects of resistin on insulin signaling in endothelial cells

The objective of this study was to investigate the effects of resistin on insulin signaling in human umbilical vein endothelial cells(HUVECs).HUVECs were incubated with recombinant human resistin(0–100 ng/mL)for 24 h.Akt and endothelial nitric oxide synthase(eNOS)phosphorylation levels of endothelial cells under basal or insulin stimulated conditions were measured by Western blot.Nitric oxide(NO)production of HUVECs was also detected.The results showed that resistin could significantly inhibit Akt and eNOS phos-phorylation and NO production in endothelial cells under insulin stimulated conditions(P<0.05 vs control).But under basal conditions,treatment with resistin could result in a decrease in eNOS phosphorylation(P<0.05 vs control)but had no effect on NO production and Akt phosphorylation levels.Thesefindings suggested that resistin exerted an inhibitory effect on NO production by inhibiting insulin signaling and eNOS phosphorylation in endothelial cells.

Developmental expression of CAPON and Dexras1 in spinal cord of rats

To study the expression of the carboxy-ter-minal PSD-95/DLG/ZO-1 ligand of nNOS(CAPON)and Dexras1 mRNA during development in the spinal cord of rats,real-time polymerase chain reaction(Real-time PCR),as a quantitative method,was used to study the developmental expression of CAPON and Dexras1 mRNA level in the spinal cord.The spatial expression of CAPON and Dexras1 mRNA was examined by a com-bination of in situ hybridization(ISH)and immunofluor-escence.During the development of the spinal cord,CAPON mRNA was expressed in low levels from embryo day 14 to day 18.At postnatal day 1,it reached the peak and was expressed in the part which will become the dor-sal horn when mature.It then decreased gradually until postnatal week 12,when it presented in the ventral horn.At embryo day 14,Dexras1 mRNA was expressed at low levels,increased during embryo day 16 to day 18 and peaked at postnatal day 1.Spatiotemporal expression of Dexras1 mRNA was similar to CAPON as confirmed by correlation analysis and colocalization.CAPON and neuronal nitric oxide synthase(nNOS)was expressed within the same cells of the dorsal horn at postnatal day 1 but had different subcellular localizations.Co-express-ion of CAPON and Dexras1 mRNA in myeloid tissue during development process of rat indicates that the adaptor protein,CAPON may play a probable role in differentiation of neurons,synaptic plasticity and synap-togenesis by regulating nNOS to activate Dexras1.