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This study explored the effect and mechanism of Maiwei Yangfei Decoction(MWYF) on pulmonary fibrosis(PF) mice. MWYF was prepared, and its main components were detected by ultra-high-performance liquid chromatography-triple quadrupole tandem mass spectrometry(UPLC-MS/MS). Male C57BL/6J mice were randomly divided into a control group, a model group, a pirfenidone(PFD) group, and low-, medium-, and high-dose MWYF groups, with 10 mice in each group. The PF model was induced in mice except for those in the control group by intratracheal instillation of bleomycin(BLM), and model mice were treated with saline or MWYF or PFD by gavage the next day. The water consumption, food intake, hair, and activity of mice were observed daily. The pathological changes in lung tissues were observed by hematoxylin-eosin(HE) staining, Masson staining, and CT scanning. The level of hydroxyproline(HYP) in lung tissues was detected by alkaline hydrolysis. Immunohistochemistry was used to observe the expression of collagen type Ⅲ(COL3) and fibronectin. The mRNA expression levels of α-smooth muscle actin(α-SMA), type Ⅰ collagen α1(COL1α1), COL3, and vimentin were detected by reverse transcription real-time fluorescence quantitative polymerase chain reaction(RT-qPCR). Superoxide dismutase(SOD) and malondialdehyde(MDA) kits were used to detect oxidative stress indicators in lung tissues and serum. The nuclear translocation of nuclear factor E2-related factor 2(Nrf2) protein was detected by immunofluorescence. The protein and mRNA expression levels of Nrf2, catalase(CAT), and heme oxygenase 1(HO-1) in lung tissues were detected by Western blot and RT-qPCR. Twelve chemical components were detected by UPLC-MS/MS. Animal experiments showed that MWYF could improve alveolar inflammation, collagen deposition, and fibrosis in PF mice, increase body weight of mice, and down-regulate the expression of fibrosis indexes such as HYP, α-SMA, COL1α1, COL3, fibronectin, and vimentin in lung tissues. In addition, MWYF could potentiate the activity of SOD in lung tissues and serum of PF mice, up-regulate the expression level of Nrf2, and promote its transfer to the nucleus, up-regulate the levels of downstream antioxidant target genes CAT and HO-1, and then reduce the accumulation of lipid metabolite MDA. In summary, MWYF can significantly improve the pathological damage and fibrosis of lung tissues in PF mice, and its mechanism may be related to the activation of the Nrf2 pathway to regulate oxidative stress.
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Camundongos , Masculino , Animais , Fibrose Pulmonar/induzido quimicamente , Fator 2 Relacionado a NF-E2/metabolismo , Fibronectinas/metabolismo , Vimentina/metabolismo , Cromatografia Líquida , Camundongos Endogâmicos C57BL , Espectrometria de Massas em Tandem , Estresse Oxidativo , Superóxido Dismutase/metabolismo , RNA Mensageiro/metabolismoRESUMO
Objective To explore the smoking and smoking cessation status in patients with acute myocardial infarction.Methods 456 hospitalized patients with acute myocardial infarction in Xicheng district were recorded in CCU ward between October 2003 and October 2008.Personal data and smoking status were collected.The smoking cessation status after discharge was investigated by telephone.Results (1) Patients who smoked were still male-dominated (96.3%).The average smoking rate in male patients was 55.9%,and even as high as 87.5% in patients at 29-50 years of age.(2) The average age in patients who smoked and with acute myocardial infarction was 58.0±12.3 years old,16 years advanced the age compared to the groups who never smoked or after stopped smoking.(3) The successful smoking cessation rate in patients with acute myocardial infarction after discharge was 42.5%,and 29-50 years old group having the highest rate of successful cessation,while the lowest rate seen in 51-65 years old group.(4) The failure rate of smoking cessation was 40.9% with the main reasons as:radical habit on smoking,withdrawal symptoms,stress in work and peer influence etc.The 51-65 year-old group was mainly suffered from habitual factors and withdrawal symptoms.Conclusion The smoking rate and smoking cessation failure rate in adult patients with acute myocardial infarction in Xicheng district in Beijing remained high.The onset age of acute myocardial infarction was significantly in advance among patients who smoked.To actively advocate on smoking cessation was still vital for reducing the occurrence of acute myocardial infarction and to improve the prognosis in patients with myocardial infarction.
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<p><b>OBJECTIVE</b>To investigate the effect of miR-122 on the expression of hepatitis B virus (HBV) proteins.</p><p><b>METHODS</b>Anti-sense oligodeoxynucleotide (ASODN) of two different sequences against miR-122, anti-miR-122 and LNA-antimiR-122 (Locked nucleic acid), human miR -122 (hsa-miR-122), or the negative control anti-GFP were designed and synthesized, then transfected into HepG2.2.15 cells. After 24 h and 48 h, the levels of HBsAg and HBeAg in the supernatant were determined with a time-resolved immunofluorometric assay (TRFIA). HBV DNA in supernatant and miR-122 in cells were measured by quantitative real-time PCR.</p><p><b>RESULTS</b>After 48 h expressions of miR-122 in the LNA-antimiR-122 and anti-miR-122 groups were significantly suppressed and lower than those in the negative control (P<0.001), while the level of miR-122 in the hsa-miR-122 group was higher than that in the negative control (P<0.001). The expression of HBeAg and HBsAg in hsa-miR-122 group was lower than that in the negative control (P<0.01) 24 h and 48 h after transfection. The expression of HBeAg and HBsAg in the anti-miR-122 group and LNA-antimiR-122 group was significantly lower than that in negative control (P>0.001). The levels of viral DNA at both time-points in the various test groups were not significantly different from those of negative control group (P>0.05).</p><p><b>CONCLUSION</b>miR-122 may regulate HBV antigens and potentially affect the progress of pathogenesis, which might be the new targets for treatment of HBV infection.</p>
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Humanos , DNA Viral , Genética , Células Hep G2 , Antígenos de Superfície da Hepatite B , Metabolismo , Antígenos E da Hepatite B , Metabolismo , Vírus da Hepatite B , Genética , MicroRNAs , Genética , Metabolismo , TransfecçãoRESUMO
OBJECTIVE@#To explore the effect on DNA quantification and STR typing from cigarette butts collected at different time points.@*METHODS@#Forty "Hongshuangxi" brand cigarette butts smoked by ten different individuals (4 cigarettes per individual) were collected. DNA was extracted from the outer layer and the sponge of the cigarette butts using chelex-100 extraction kit, as well as STR typing and DNA quantitation were simultaneously performed in 1, 4, 7 and 10 weeks, respectively.@*RESULTS@#The DNA quantities extracted from the outer layer at the 1st, 4th, 7th and 10th week were 0.104-2.52, 0.110-2.41, 0.0960-2.32 and 0.085 0-2.28 ng/microL, while the detection rates for 16 loci by STR typing were 100%, 90%, 75% and 62.5%, respectively. The DNA quantities extracted from the sponge were 0.0180-2.40, 0.0171-2.25, 0.0165-2.15 and 0.0160-2.15 ng/microL, while the detection rates for 16 loci by STR typing were 97.5%, 82.5%, 50% and 12.5%, respectively.@*CONCLUSION@#There is little difference in DNA quantity between the outer layer and the sponge of butts during 10 weeks, but there is an obvious effect on STR typing with prolonged extracting time. There is a much more effect on the sponge than on the outer layer, and the longer the standing time is, the lower the detection rate is.
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Humanos , DNA/análise , Impressões Digitais de DNA/métodos , Células Epiteliais/química , Genética Forense/métodos , Repetições de Microssatélites/genética , Mucosa Bucal/citologia , Fumar , Fatores de TempoRESUMO
<p><b>OBJECTIVE</b>To evaluate the inhibition effects of DNAzymes specific to Hepatitis B Virus(HBV) s gene and e gene on the expressions of Hepatitis B surface antigen(HBsAg) and Hepatitis B e antigen(HBeAg).</p><p><b>METHODS</b>DNAzymes DrzBS and DrzBC specific to HBV s gene ORF A157UG and e gene ORF A1816UG, respectively, were designed and synthesized. The inhibition effects of DrzBS or DrzBC on the expressions of HBV s and e genes were observed in 2.2.15 cells.</p><p><b>RESULTS</b>The expression of HBV s or e genes was dramatically depressed after 2.2.15 cells treated by DrzBS or DrzBC. The concentration for effective inhibition was within 0.1-2.5 micromol/L and the inhibition showed a dose dependence within that concentration range. The maximum inhibition was 94.2% and 91.8% for DrzBS and DrzBC, respectively. The inhibition was maintained for 72 hours. The efficiency of inhibiting HbsAg, HbeAg in 2.2.15 cells by DrzBS, DrzBC was higher than that by antisense oligonucleotides for the same target genes. The concentrations for effective inhibition of the DNAzymes were at least 10-fold lower compared with antisense oligonucleotides. Neither inhibition on the replication of HBV DNA nor toxicity to 2.2.15 cells was observed.</p><p><b>CONCLUSION</b>DrzBS and DrzBC can highly block the expressions of HBV s gene and e gene in 2.2.15 HBV cell model and are proved a specific and effective anti-HBV gene therapeutic means.</p>