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@#To investigate the effects of intratracheal instillation of PM2.5 suspension on bleomycin (BLM)-induced pulmonary fibrosis in mice and the intervention of neotuberostemonine (NTS), the BLM dose (1.5 or 3.0 U/kg) and PM2.5 frequency (1 or 2 times per week) were studied by factorial experiment design. After intratracheal instillation of BLM (1.5 or 3.0 U/kg) on day 0, PM2.5 (5 mg/kg) was intratracheally injected to mice once or twice a week from day 1 to day 21, and the mice in the treatment group were given 30 mg/kg NTS by gavage once a day from day 8 to day 21. The degree of pulmonary fibrosis was evaluated by lung coefficient, hydroxyproline (HYP) content, HE staining and Masson staining lung sections as well as their semi-quantitative index (HE inflammatory score and collagen volume fraction, CVF). The results showed that the HE scores increased significantly in mice singly given PM2.5 once a week, the HYP content and HE score increased in mice singly given PM2.5 twice a week, but their CVF values did not significantly increase. However, the CVF values increased significantly in mice treated with PM2.5 and BLM co-infusion. These results suggested that PM2.5 (administered singly) could significantly increase BLM-induced collagen deposition and greatly aggravate pulmonary fibrosis although it mainly caused pulmonary inflammation rather than pulmonary fibrosis. NTS could significantly reduce the CVF value and α-SMA protein level of the model mice. It can be concluded that PM2.5 has great influence on patients with respiratory diseases, while NTS can improve pulmonary fibrosis induced by the combination of PM2.5 and BLM.
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@#This study aimed to investigate the protective effects of Baibu Tang on bleomycin-induced pulmonary fibrosis in mice. After intratracheally giving bleomycin(3. 5 mg/kg), mice were orally administered Baibu Tang once a day for 14 consecutive days, takingnintedanib as a positive control. The anti-fibrotic effects were assessed by the hydroxyproline level and the histopathological changes in H&E or Masson stained lung tissues. The results revealed that the lung coefficient, hydroxyproline content, inflammation and collagen deposition were increased significantly in the lung tissue of the model mice. Both ethanol and water extracts of Baibu Tang significantly improved all the pathological indexes in mice, but the effect of the ethanol extract was better than that of the water extract. Baibu Tang with Baibu(root of Stemona tuberosa)containing different components(neotuberostemonine, tuberostemonine and stemoninine, respectively)could significantly reduce hydroxyproline level and collagen deposition in the lung tissue of bleomycin-induced mice, and there was no significant difference in their activity. This result showed that the changes in the chemical composition of Stemona tuberosa(Baibu, monarch drug for Baibu Tang)have little effect on the anti-fibrosis activity of Baibu Tang, and its mechanism and material basis need further investigation.
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@#This study aimed to investigate the protective effect of total alkaloids(TA)and one of the active components, neotuberostemonine, of Stemona tuberosa on bleomycin-induced pulmonary fibrosis in mice and to explore the mechanism by fibroblasts model. The bleomycin-induced mice were orally administered with TA(60 mg/kg)and neotuberostemonine(10, 20 mg/kg), with prednisone(6. 67 mg/kg)as a positive control. The anti-fibrotic effects were assessed by hydroxyproline content, TGF-β1 level, inflammatory score, collagen deposition and the expression of α-SMA in the lung tissues. The results revealed that TA and neotuberostemonine could significantly ameliorate the inflammation and injury, and attenuate the hydroxyproline content and collagen deposition. Moreover, neotuberostemonine treatment markedly down-regulated the α-SMA level and TGF-β1 content in bleomycin-injured mice lungs. The in vitro experiments showed that neotuberostemonine inhibited the expression of α-SMA induced by TGF-β1 in a dose-dependent manner, indicating that suppression on differentiation of fibroblasts to myofibroblasts may be one of the mechanisms for neotuberostemonine against pulmonary fibrosis.
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Objective To explore the application effect of grommet insertion and adenoidectomy application effect in the treatment of children with secretory otitis media.Methods 64 children with secretory otitis media were randomly divided into two groups,32 cases in the observation group,32 cases in the control group.The control group used a simple tympanic membrane in the treatment of catheter,the observation group was treated with grommet insertion and adenoidectomy,follow-up 6 months curative effect were observed.Results The control group,the observation group after treatment backbone transmission difference were (12.1 ± 1.9)dB,(4.5 ± 1.2)db,two groups of backbone transmission errors were significantly lower than that before treatment (t =4.432,5.357,all P ＜ 0.05),compared with the control group,the observation group was significantly lower (t =10.299,P ＜ 0.05).The control group,the observation group of eustachian tube function recovery rate were 25.0%,68.8%,there was significant difference between two groups (x2 =3.96,P ＜ 0.05).Total effective rate in the observation group was 93.8 %,which was higher than 71.9% in the control group(x2 =5.42,P ＜ 0.05).Conclusion Tympanic membrane and adenoidectomy in the treatment of children with secretoryotitis media has significant effect.
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Objective To obtain recombinant human granulysin using prokaryotic expression system. Methods Total RNA was extracted from cultured PBMC. Granulysin gene segments were obtained with granulysin-specific primers by RT-PCR and then inserted into pET32a(+) plasmid. After identification by DNA sequence, pET-GN-LY9K and pET-GNLY15K were transferred to E. Coli Rosetta (DE3). The fusion protein was identified by SDS-PAGE and Western blot. The bioactivity of granulysin fusion protein was measured by MTT assay. Results The prokaryotic expression vectors pET-GNLY9K and pET-GNLY15K were successfully constructed. The corresponding protein was highly expressed in E. Coli. Recombinant protein was specifically bound by anti-granulysin antibody. GNLY9K fusion protein significantly inhibited the growth of A549 cells in a dose-dependent manner, while GN-LY15K had little effect on the growth of A549. Conclusion Granulysins with different mw were successfully expressed using prokaryotic expression system, which might be helpful for the further study of granulysin.
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Objective The effectiveness on the enrichment of nuclear red blood cells(NRBC) in umbilical cord blood by polyvinylidene difluoride membrane was investigated with intervention on membrane ionic channels.Methods NRBC was isolated by the isolation solution with a density of 1.067 g/ml.The rates of the cell enrichment using verapamil and furosemide intervention by discontinuous density centrifugation were examined by flow cytometry.The NRBC that past through PVDF membrane with pore size of 5 ?m were checked and quantified by nested-PCR.Results The optimal centrifugation density was 1.067 g/ml.The mean purity of NRBC after centrifugation was 2.54%,yet the purity after intervention increased to 9.36%(3.7-fold increment).The NRBC passing through PVDF membrane was from 0.83% to 6.15%,with enrichment of 7.4 times.When NRBC was 120 cells/ml,the density of the nested PCR products increased by nearly 4 times after intervention(4.48 vs 17.78).Conclusions Verapamil and furosemide could affect the morphology of cells and promote the rate of NRBC enrichment.
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Objective To obtain recombinant human granulysin using prokaryotic expression system.MethodsTotal RNA was extracted from cultured PBMC. Granulysin gene segments were obtained with granulysin-specific primers by RT-PCR and then inserted into pET32a(+) plasmid. After identification by DNA sequence,pET-GNLY9K and pET-GNLY15K were transferred to E. coli Rosetta (DE3). The fusion protein was identified by SDS-PAGE and Western blot.The bioactivity of granulysin fusion protein was measured by MTT assay.Results The prokaryotic expression vectors pET-GNLY9K and pET-GNLY15K were successfully constructed.The corresponding protein was highly expressed in E.coli. Recombinant protein was specifically bound by anti-granulysin antibody. GNLY9K fusion protein significantly inhibited the growth of A549 cells in a dose-dependent manner,while GNLY15K had little effect on the growth of A549.Conclusion Granulysins with different mw were successfully expressed using prokaryotic expression system,which might be helpful for the further study of granulysin.
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OBJECTIVE To observe the clinical characteristics and risk factors of the hospital-acquired infection among uremia patients.METHODS The clinical data of 187 cases of uremia were retrospectivety analyzed during from Apr 2003 to Dec 2006.RESULTS There were 61 cases with hospital-acquired infection.The infection rate was 32.6%.The repiratory(54.1%) and urinary(21.3%) systems were the main infection sites.The main pathogens were Escherichia coli,Klebsiella pneumoniae,Staphylococcus epidermidis,S.aureus and fungi.The related factors were renal function,serum albumin,hemoglobin,duration in hospital,age and invasive medical procedures.CONCLUSIONS The hospital-acquired infection with uremia is with higher rate in respiratory system.The risk factors are renal failure,decrease in serum albumin and hemoglobin,abuse use of antibiotics and invasive medical procedures and deserve corresponding nursing strategy.