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AIM:To explore the relationship and molecular mechanism between microRNA-21(miR-21) and Schwann cells ( SC) following peripheral nerve injury.METHODS: The mRNA expression of miR-21 and phosphatase and tensin homologue deleted on chromosome ten ( PTEN) in animal model were detected by real-time PCR.The over-ex-pression of miR-21 and inhibition of miR-21 expression in the Schwann cells according to transfection of lentiviral vectors were performed, the nonspecific miRNA was used as a negative control ( NC) .The cell apoptosis was measured by flow cy-tometry.The mRNA expression of miR-21 and PTEN in the cells was detected by real-time PCR.The protein expression of PTEN and cleaved caspase-3 was determined by Western blot.RESULTS: The level of miR-21 was significantly higher and the mRNA level of PTEN was significantly lower in the model of nerve injury than those in control group.miR-21 over-expression decreased the number of apoptotic Schwann cells compared with NC-SC.The mRNA expression of PTEN was down-regulated by over-expression of miR-21.The protein expression of PTEN and cleaved caspase-3 was down-regulated by over-expression of miR-21 (P<0.05).CONCLUSION: miR-21 may play an important role in the peripheral nerve injury through inhibiting apoptosis of Schwann cells by down-regulating the expression of PTEN.
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AIM: To investigate the molecular mechanism of microRNA-21 (miR-21) in the regulation of Schwann cell proliferation following nerve injury.METHODS:The expression of miR-2l was detected by real-time PCR. Synthetic miR-21 mimic and its control were transfected into rat Schwann cells.CCK-8 assay was performed to evaluate the influence of miR-21 on the proliferation of Schwann cells.The cell cycle distribution was determined by flow cytometry. The expression of transforming growth factorβ-induced protein ( TGFBI) and cyclin D1 were detected by Western blotting. RESULTS:The expression of miR-21 in model group was 7.87 ±0.75 and 7.75 ±0.80 times higher than that in sham operation group and blank group respectively.After transfected with miR-21 mimic, the expression of miR-21 in experimen-tal group was 2.21 ±0.14 and 2.29 ±0.21 times higher than that in negative control group and blank group respectively. Moreover, the A450 value of CCK-8 assay in experimental group at 48 h was higher than that in negative control group and blank group.The proliferation index in experimental group was higher than that in negative control group and blank group. At the same time, the expression of TGFBI obviously decreased and the cyclin D1 increased in the Schwann cells 48 h after transfection with miR-21.CONCLUSION:miR-21 promotes the proliferation activity of Schwann cells by down-regulating TGFBI expression.
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Aim To study the inhibitory effects of ex-tractive pericarpium trichosanthes ( EPT) on high glu-cose-induced apoptosis in human umbilical vein endo-thelial cells ( HUVECs ) and its underlining mecha-nisms. Methods HUVECs were cultured. Effects of EPT at different concentrations on the high-glucose-in-duced apoptosis in HUVECs were observed. The cell viability of HUVECs was determined by MTT colori-metric method. Cell apoptosis was identified by Ho-echst staining. The intracellular activity of Caspase-3 was detected with colorimetry. Protein expression and p65 nuclear translocation of NF-κB were detected by Western blot and immunofluorescence staining. Re-sults Treated with 30 mmol · L-1 glucose for 48 hours had a significantly decrease on cell viability com-pared with control, and the apoptotic rate and Caspase-3 activity were increased markedly, the protein expres-sion of NF-κB was upregulated and p65 nuclear trans-location in HUVECs increased; Pre-incubation with EPT(12. 5,25,50 mg·L-1 ) for 1 hour enhanced the cell viability, and decreased the apoptotic rate and Caspase-3 activity and downregulated the expression of NF-κB in a dose-dependent manner. Moreover, EPT could inhibit p65 translocation. Conclusion EPT can protect HUVECs against the apoptosis induced by high glucose in vitro,and its mechanism may be related with downregulation of NF-κB expression and inhibition of the intracellular activity of Caspase-3 .
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ObjectiveTo observe the therapeutic effect and recurrence of treating Benign Paroxysmal Positional Vertigo(BPPV) by the treatment that combined Canalith Repositioning Maneuver(CRM) with Chinese and Western medicine.MethodsCollecting 40 cases suffered from BPPV,and randomized controlled method was used.Control group:20 cases( using Western and Chinese medicine),treatment group:20cases( using Western and Chinese medicine wrbined with CRM),judge the effect after 7days and telephone followed up 3 months,inquiring about recurrence situation.ResultsCure rate of treatment group was 90％,total effective rate was 100％ which was higher than the control group which was 50％,the total effective rate was 95％ (P ＜0.001 ) ;20 cases in treatment group followed up by telephone didn't recur.1 case in control group recurred,but got better after the treatment of CRM.ConclusionCRM which was effective,safe,simple and convenient to treat BPPV could be used as the preferred treatment for BPPV,if combined with Chinese and western medicine,it could significantly alleviate the symptoms and reduce the relapse.
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To study the chemical constituents of Bauhinia glauca subsp. pernervosa, eleven phenolic acids were isolated from a 95% ethanol extract by using a combination of various chromatographic techniques including column chromatography over silica gel, ODS, MCI, Sephadex LH-20, and semi-preparative HPLC. By spectroscopic techniques including 1H NMR, 13C NMR, 2D NMR, and HR-ESI-MS, these compounds were identified as isopropyl O-beta-(6'-O-galloyl)-glucopyranoside (1), ethyl O-beta-(6'-O-galloyl)-glucopyranoside (2), 3, 4, 5-trimethoxyphenyl-(6'-O-galloyl)-O-beta-D-glucopyranoside (3), 3, 4, 5-trimethoxyphenyl-beta-D-glucopyranoside (4), gallic acid (5), methyl gallate (6), ethyl gallate (7), protocatechuic acid (8), 3, 5-dimethoxy-4-hydroxybenzoic acid (9), erigeside C (10) and glucosyringic acid (11). Among them, compound 1 is a new polyhydroxyl compound; compounds 2, 10, and 11 were isolated from the genus Bauhinia for the first time, and the other compounds were isolated from the plant for the first time. Compounds 6 and 8 showed significant protein tyrosine phosphatase1B (PTP1B) inhibitory activity in vitro with the IC50 values of 72.3 and 54.1 micromol x L(-1), respectively.
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Estrogen Receptor (ERalpha) is a member of superfamily of ligand-activated transcription factors which play critical roles in many biological processes. To screen novel modulators of ERalpha for drug development and biological function research, we developed a mammalian one-hybrid-based high-throughput screening model for ERalpha modulator. We cloned the ERalpha LBD gene from the total mRNA of fat tissue by RT-PCR and fused it with the GAL4 DNA binding domain of pBIND-GAL4 plasmid to construct a chimara expression plasmid pBIND-GAL4-Eralpha(LBD). The L02 cells was cotransfected with pBIND-GAL4-ERalpha(LBD) and a GAL4-responsive luciferase reporter plasmid pGL3-GAL4, and following treatment with test compounds for 24 h, the activities of luciferase were detected to evaluate the transactivities of ERalpha modulators. After manner optimizations of transfection conditions, Estradiol, an agonist control, induced the expression of luciferase in a dose-dependent with EC50 of 0.17 micromol/L, the maximum folds of induction was about 28.1. Tamoxifen, an antagonist control, efficiently suppressed the estradiol-mediated luciferase induction with EC50 of 0.10 micromol/L. Using this screening model, we discovered four ERalpha agonists from 2000 natural and synthetic compounds.
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Animals , Humans , Mice , 3T3-L1 Cells , Chimera , Metabolism , DNA-Binding Proteins , Genetics , Estrogen Receptor Modulators , Chemistry , Estrogen Receptor alpha , Genes, Reporter , Genetics , Genistein , Chemistry , HeLa Cells , Luciferases , Genetics , Metabolism , Models, Chemical , Saccharomyces cerevisiae Proteins , Genetics , Transcription Factors , Genetics , TransfectionABSTRACT
Objective To assess the relationship between position of incisor and soft tissues changes in class Ⅲ malocclusion cases after orthodontic treatment.Methods All 26 patients in this study were diagnosed as skeletal class Ⅲ malocclusion.The lateral X-ray cephalometric films taken at the beginning and the end of the treatment were analyzed for position of incisor and soft tissues changes.Statistical analysis Waft performed using SAS v8.0 software.Resuits There was correlation between the labial inelination of upper and lower incisors and the soft tissue profile.Conclusion Soft tissue will be harmonious by the labial inclination increase of the upper incisor teeth and reduction of the lower incisor teeth.
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Objective： To clone rat cardiac myosin α he avy chain cDNA fragment encoding aa736-960 and construct its recombinant retrov irus vector(RV). Methods: The 681 bp target gene was amplified f rom heart tissue of young rats with RT-PCR, fusion gene of huIL-2/myosin was c onstructed by splicing with huIL-2 cDNA using ligation methods and its RV was constructed. RT-PCR and immunohistochemical assay were used to iden tify the expression of myosin protein in transfected cell. Results: The determination of nucleotide sequence showed that the nucleotide and ami no acid sequence of gene clone was the same as reported, its openin g reading frame was correct, the digesting result of pLNC-huIL-2-myosin was i dentical with the predicted. NIH3T3 cell was transfected with recombinant RV, and G418-resistant NIH3T3 cell was established.RT-PCR analysis indicated tha t mRNA of pLNC-huIL-2-myosin was present in cell transfected with RV. The im munohistochemical assay also showed that the myosin protein expression was highe r in the cell transfected with constructed RV. Conclusion: Rat cardiac myosin α heavy chain cDNA has been cloned and its RV has also been cons tructed and expressed in NIH3T3 successfully, it will contribute to research of prevention and treatment of heart immune injury by cardiac myosin gene transfer to induce specific tolerance.
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Objective To study the effect of volatile chemicals in the air of newly decorated rooms on O6-methylguanine-DNA methyltransferase (MGMT) expression in the lung, liver, kidney and testicle tissues of mice. Methods Forty Kunming mice were randomly divided into four groups and the imitate pollutants mixture of volatile chemicals in the air of newly decorated rooms was used as the exposed toxicants: low-dose exposure group (7.5 mg/m3), moderate-dose exposure group (15 mg/m3), high-dose exposure group (30 mg/m3), the control group (no exposure ). The mice were exposed 4 hours each day, for nine weeks. Streptavidin and biotechnology-peroxidase (SABC) immunohistochemical method was used to determine the MGMT expression in the lung, liver, testicle and kidney tissues. Results The expression of MGMT in the liver, lung, testicle tissues in the high, moderate and low-dose groups reduced significantly compared with the control group (P0.05). Conclusion The higher exposure of volatile chemicals from the air of newly decorated rooms may inhibit the MGMT expression in the liver, lung and testicle tissues of mice.