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OBJECTIVE Glioblastoma multiforme (GBM) is the most malignant primary tumor of the central nervous system and is associated with a very poor prognosis. No further improvements in outcomes have been reported since radiotherapy-temozolomide therapy was introduced.Therefore,de-veloping new agents to treat GBM is important. This study aimed to evaluate the anti-tumor effect of evodiamine (Evo) on GBM cells, and to determine the underlying mechanisms involved. METHODS U251,LN229,HEB and PC12 cells were treated with various concentrations of evodiamine for 24 and 48 hours,cell viability was measured by MTT assay.The U251 and LN229 cells were treated with evo-diamine(0-10 μmol·L-1)for 24 h,and then stained with Hoechst 33258.An Annexin V-FITC Apoptosis Detection Kit was used to detect apoptosis in the cells.Reactive oxygen species(ROS)production was detected using dichlorofluorescein diacetate (DCFH-DA) staining. The changes in mitochondrial mem-brane potential (MMP) were assessed by JC-1 after cells were treated with evodiamine. The expres-sion levels of p-PI3K,PI3K,p-Akt,Akt,Bax,Bcl-2,p-p38,p38,p-JNK,JNK,p-ERK,ERK,Cytochrome c, Caspase-3, cleaved Caspase-3, PRAP, and cleaved PARP were measured by Western blot analy-ses. RESULTS According to MTT assay results, Evo significantly inhibited the cell proliferation in a time- and dose-dependent manner. Fluorescence microscopy and flow cytometry analyses revealed that Evo induced cell apoptosis in a concentration-dependent manner.Moreover,Evo induced reactive oxygen species (ROS) production and mitochondrial membrane potential (MMP) disruption. Finally, Evo induced apoptosis in cancer cells by suppressing PI3K/AKT signaling and inducing MAPK phos-phorylation(p38 and JNK,but not ERK)to regulate apoptotic proteins(Bax,Bcl-2,Cytochrome c,Cas-pase-3, and PARP). CONCLUSION In summary, Evo inhibits cell proliferation by inducing cellular apoptosis via suppressing PI3K/AKT and activating MAPK in GBM;these results indicate that Evo may be regarded as a new approach for GBM treatment.
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AIM:To explore the characteristics of hepatitis B virus S gene mutation in the vertical transmission after active and passive vaccination .METHODS:Fifteen cases of immunoprophylaxis failure were enrolled in the study . HBV S gene (including pres-S and S) from the mothers, newborns before active and passive vaccination and 7-month-old infants with immunoprophylaxis failure were detected by PCR amplification .The characteristics of HBV S gene mutation were compared among the 3 groups.RESULTS: The genotype of HBV in the newborns and the infants was the same as that in the mothers .The frequencies of mutation in the 2 fragments of the HBV S gene had no significant difference between the 3 groups.The homology tree model based on HBV S gene was analyzed in the 3 groups, in which every group had their own cluster.There were 15 different mutation sites between 7 pairs of mothers and newborns .There were 3 different muta-tion sites between 3 pairs of newborns and infants (nt273A→A/G, nt512C→C/T and nt1139C→A), among which the first 2 were located in the S gene region but not in the “a” determinant , and the latter was located in the overlap region of S and X genes .There were 25 different mutation sites between 9 pairs of mothers and infants , but only 1 case had a differ-ent mutation site between the mother , newborn and infant .CONCLUSION: The HBV species in newborns and infants with immunoprophylaxis failure were transmitted from the mothers .The mutations in the HBV S gene with immunoprophy-laxis failure happened before and after active and passive vaccination , mainly before vaccination .The relationship between HBV S gene mutations and immunoprophylaxis failure should be further explored .
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The training of clinical thinking ability is the key point of obstetrical teaching to medical students in eight-year program.Obstetrical department of the third affiliated hospital of Sun Yat-sen university has put forward specific methods of SP inquiring and PBL case analysis teaching.The well-designed teaching methods have an active role in the training of case history collecting ability,diagnosing and treating ability to medical students in eight-year program.
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<p><b>OBJECTIVE</b>The purpose of this study was to clone and analyze mutation in the eda-A1 gene for hypohidrotic ectodermal dysplasia (HED), and to construct a new recombined eukaryotic expression vector (mutant M, wild W) as a basis for further study on the genetic function.</p><p><b>METHODS</b>After total mRNA was extracted from peripheral blood lymphocytes from the HED affect patient and control, eda-A1 gene was amplified by reverse transcription polymerase chain reaction (RT-PCR) with a pair of specific primers containing the constriction enzyme sites of BamH I and Hind III. When the vector pcDNA3.1(-) and eda-A1 (M/W) were digested by BamH I and Hind III respectively, eda-A1 (M/W) fragment was then ligated to vector pcDNA3.1 (-) and the new vector was named as pcDNA3.1 (-)-eda-A1-M/W.</p><p><b>RESULTS</b>eda-A1 gene was successfully cloned and a novel missence mutation was identified, which changes the codon 306 from glutamine to proline. PCR, restrictive endonuclease analysis and DNA sequencing were then performed to identify the recombinant eukaryotic expression vector pcDNA3.1 (-)-eda-A1-M/W, and the results were surely confirmed.</p><p><b>CONCLUSION</b>Our result indicates that the novel missense mutation in eda is associated with the isolated tooth agenesis and provide preliminary explanation for the abnormal clinical phenotype at a molecular structural level. And also, the recombinant eukaryotic expression vector pcDNA3.1 (-)-eda-A1-M/W was successfully constructed, which will be thereafter taken use of further study on eda gene in odontogenesis.</p>
Subject(s)
Humans , Ectodermal Dysplasia 1, Anhidrotic , Genetic Vectors , Mutation , Odontogenesis , RNA, Messenger , Sequence Analysis, DNA摘要
Objective: To investigate the candidal infection status in puerperas in Lanzhou, and the candidal transmission from mothers to their newborn infants. Methods: Vaginal fluid and saliva samples from 104 puerperas, as well as 104 saliva samples from their newborn infants were collected. The Candida species were cultured, isolated and identified using CHROMagar media. Further identification was done using molecular biological method. Results; In 81 of 312 specimens (104 x2 from mothers and 104 from infants), Candida species were found. 39.42% (41 cases) was observed in the vaginal fluid and 33.65% (35 cases) was in saliva of puerperas respectively, and 21. 15% (22 cases) in both vagina and oral cavity. 4.81% (5 cases) was found in oral cavities of newborn infants. The distribution of Candida species were 53 Candida albicans, 33 Candida glabrata, 2 Candida krusei and 1 Candida tropical. In 2 pairs of mother-infant, the same genotype of Candida ablicans was identified using PCR method. Conclusion; The Candida detection rate of newborn infants and transmission rate from mothers to their neonates in Lanzhou are higher than that reported in other areas. The colonization of Candida in newborn infants is relevant to both horizontal and vertical transmission. It can decrease the possibility of Candidal infection in newborn infants by controlling the Candidal transmission in hospital and preventing the infection in pregnant women.
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Astragalus produced in Gansu were chosen as the raw material to leachate. Studied the antibiotic effects of the leaching solution on the cariogenic bacteria and compared with the imported bacteriostatic product MI. Streptococcus mutans and Lactobacilli were cultured in the medium for 24 h. The PH and A600 values were measured. Statistical analysis was conducted by using SPSS 13.0. The leaching solution of astragalus has the same inhibitory effects on the growth and acid production of streptococcus mutans and lactobacilli as MI.
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<p><b>OBJECTIVE</b>To evaluate the effect and related mechanisms of aldosterone (ALD) on inducible nitric oxide synthase (iNOS) activity and nitric oxide (NO) production in aortic adventitia.</p><p><b>METHODS</b>Aortic adventitias from SD rats were incubated for 6 hours with various protocols: buffer alone (control), ALD (10(-8) mol/L - 10(-6) mol/L), ALD + spironolactone (10(-5) mol/L, ALD + SP), ALD + RU486 (10(-5) mol/L), LPS 10 ng/ml (LPS), ALD + LPS (10 ng/ml), ALD + LPS + SP (10(-5) mol/L), and ALD + LPS + RU486. Nitrate/nitrite (NOx), an index of NO production, was measured by Greiss Reaction. iNOS activity was determined by isotope-labeled L-arginine convertion rate.</p><p><b>RESULTS</b>(1) NOx production and iNOS activity were similar between ALD and control groups (P > 0.05). NOx production was significantly reduced while iNOS activity remained unchanged in the ALD (10(-6) mol/L) + SP group compared to ALD (10(-6) mol/L) group. NOx production by 10(-7) mol/L and 10(-6) mol/L ALD increased by 50.0% and 58.7% respectively (P < 0.01) and iNOS activity was also significantly increased (P < 0.01) in ALD + RU486 group than that in ALD group. (2) LPS significantly increased the NOx production and iNOS activity (P < 0.01) and these effects were not augmented by adding ALD to LPS (P > 0.05) and SP significantly blocked and RU486 significantly enhanced the effects by LSP and ALD on NOx production and iNOS activity (P < 0.05).</p><p><b>CONCLUSION</b>Aldosterone has a dual effect on iNOS/NO through mineralocorticoid receptor and glucocorticoid receptor pathway.</p>
Subject(s)
Animals , Male , Rats , Aldosterone , Pharmacology , Aorta, Thoracic , Metabolism , Cells, Cultured , Connective Tissue , Metabolism , Nitric Oxide , Metabolism , Nitric Oxide Synthase Type II , Metabolism , Rats, Sprague-Dawley摘要
We treated 40 cases of stubborn constipation by puncturing Zhongwan (CV 12),Qihai (CV 6), Tianshu (ST 25, bilateral), Daheng (SP 15, bilateral). After 20 times of treatment, 16 cases were cured(accounting for 45%) ,18 cases were significantly improved(accounting for 45%),4 case were improved(accounting for 10%),2 cases were ineffective(accounting for 5%).The total effective rate was 95%.