الملخص
Objective To evaluate the effect of tea polyphenol epigallocatechin gallate (EGCG)on ultraviolet B (UVB)-induced skin pigmentation,transfer and degradation of melanosomes in mice,and to explore the role of autophagy in the mechanism of melanosome degradation.Methods A total of 32 ears from 16 female C57/BL6 mice were randomly and equally divided into 4 groups:acetone control group topically treated with acetone solution daily,EGCG group topically treated with 10 g/L EGCG acetone solution daily,UVB irradiation group irradiated with 500 mJ/cm2 UVB once a day and 2 hours later topically treated with acetone solution,UVB + EGCG group irradiated with 500 mJ/cm2 UVB once a day and 2 hours later topically treated with EGCG acetone solution.Ten days later,all the mice were sacrificed,and skin tissue samples were collected from the ears.Transmission electron microscopy was performed to observe ultrastructural changes of melanosomes and autophagosomes,immunohistochemical study to measure expression of protease-activated receptor 2 (PAR2) and microtubule-associated protein light chain 3 (LC3) in the epidermis,and Western blot analysis to determine the protein expression of PAR2,Rasrelated protein Rab27a and LC3 in the epidermis.Results There was a significant difference in the number of melanosomes and autophagosomes among the acetone control group,EGCG group,UVB irradiation group and UVB + EGCG group (H =12.249,13.888,respectively,both P ＜ 0.05).Compared with the acetone control group,the UVB irradiation group showed significantly increased number of melanosomes (1.85 ± 0.32 vs.1.00 ± 0.41,P ＜ 0.05)and autophagosomes (1.94 ± 0.64 vs.1.00 ± 0.46,P ＜ 0.05) in epidermal keratinocytes in mouse skin.Compared with the UVB irradiation group,the UVB + EGCG group showed significantly decreased number of melanosomes (1.30 ± 0.44,P ＜ 0.05),but significantly increased number of autophagosomes (3.03 ± 0.75,P ＜ 0.05).Immunohistochemical study showed a significant difference in the level of PAR2 in the epidermis among the 4 groups (H =18.700,P ＜ 0.05),and the expression of PAR2 was significantly lower in the UVB + EGCG group than in the UVB irradiation group (7.94 ± 4.57 vs.12.54 ± 3.07,Z =2.143,P ＜ 0.05).However,the 4 groups all showed a low level of LC3,and there was no significant difference among the 4 groups (H =5.051,P ＞ 0.05).Western blot analysis revealed significant differences in the protein expression of PAR2 and Rab27a,as well as in the LC3-Ⅱ/LC3-Ⅰ ratio,among the 4 groups (F =18.739,25.967,24.022,respectively,all P ＜ 0.05).Compared with the UVB irradiation group,the UVB + EGCG group showed significantly decreased expression of PAR2 (0.91 ± 0.54 vs.3.12 ± 0.61,P ＜ 0.05) and Rab27a (0.99 ± 0.16 vs.1.42 ± 0.07,P ＜ 0.05),but significantly increased LC3-Ⅱ/LC3-Ⅰ ratio (1.67 ± 0.08 vs.1.24 ± 0.07,P ＜ 0.05).Conclusion Topical EGCG treatment can effectively suppress UVB-induced skin pigmentation,which may be related to the inhibition of melanosome transfer and promotion of melanosome autophagy.
الملخص
AIM:To explore the effect of Delta-like ligand 4 (Dll4)-Notch signaling pathway blockade on the development of Thelper 17(Th17) cells in the asthmatic mice.METHODS:Male BALB/c mice were randomly divided into 5 groups:control group, asthma group, normal saline group, anti-Dll4 antibody group, and immunoglobulin G group.The protein expression of Dll4 was detected by immunohistochemical staining.The proportion of Th17 cells in mouse spleen isolated CD4+ T cells was measured by flow cytometry.The protein expression of Th17 transcription factor retinoid-related orphan receptor γt (RORγt) was determined by Western blot.The serum level of interleukin (IL)-17 was measured by enzyme-linked immunosorbent assay (ELISA).RESULTS:The expression of Dll4 in the lung tissues from asthma group significantly increased as compared with anti-Dll4 antibody group.The proportion of Th17 cells in CD4+ T cells was significantly down-regulated, and the protein expression of RORγt in the lung tissues was significantly reduced in anti-Dll4 antibody group compared with asthma group (P<0.05).Moreover, the serum level of IL-17 in anti-Dll4 antibody group was significantly reduced compared with asthma group (P<0.01).CONCLUSION:The blockade of Dll4-Notch signaling pathway inhibits the differentiation of Th17 cells in asthmatic mice.
الملخص
Surface enhanced Raman spectroscopy technology (SERS), using gold nanoparticles as a base, was developed for rapid and sensitive detection of virus strains. SERS can be used as a rapid and reliable method to distinguish the titers of viral replication. In the present study, we characterized H1N1 subtypes of influenza A virus strains in different conditions of pH or temperatures, while we analyzed data from SERS technology using gold nanoparticles as a base and cell cultures were employed to further confirm the data from virus strains. Origin8.0 was used to collect Raman spectra, smooth and homogenize data, and to contrast spectra. Our results indicated that the peaks of different virus strains in optimal environmental conditions (T=37 ℃/pH=7.2) reached ≥3 000. This criterion was verified by subsequent virological method. The present data indicate that the established SERS protocol can be used as a rapid and reliable method to distinguish the replication rate of virus, which can be further used in clinical samples.
الموضوعات
Gold , Hydrogen-Ion Concentration , Influenza A Virus, H1N1 Subtype , Nanoparticles , Spectrum Analysis, Raman , Temperature , Virus Cultivation , Methodsالملخص
Objective To explore the clinical manifestations,imaging findings,pathological classification and treatment of congenital cystic adenomatoid malformation (CCAM)of the lung.Methods The clinical features,imaging findings,pathology information,diagnosis,treatment method and its prognosis of children with CCAMconfirmed by ope-ration and pathology were retrospectively analyzed in Yuying Children′s Hospital Affiliated to Wenzhou Medical Univer-sity from August 2006 to August 201 4.Results Eleven patients were boys and 4 patients were girls.One case had a-symptomatic clinical features,1 2 cases had pulmonary infection,1 case had recurrent chest pain,and 1 case had de-pressed deformity in sternum inferior segment.Chest CT scanning indicated that 9 cases had multiple gas cysts at unila-teral side of lung,among which 1 case was of funnel chest and pulmonary sequestration,1 case of huge cyst containing air and fluid at inferior lobe of left lung,and 4 cases of high density lung shadow;CT examination indicated that 1 case had recurrent chest pain and eventration of diaphragm of the right side combined with pulmonary sequestration.All ca-ses were treated by surgical resection,of whom 1 case was given cystectomy and sequestrectomy,diaphragmatic plication respectively,1 case complicated with funnel chest disease underwent lesion pulmonary lobectomy,sequestrectomy and minimally invasive corrective surgery in pectus excavatum (Nuss surgery),and the remaining 1 2 cases received lesion pulmonary lobectomy.All of 1 5 cases recovered well without complications.Pathological classification type of CCAMin-cluded 1 1 cases of type Ⅰ,3 cases of type Ⅱ and 1 case of type Ⅲ,among which 2 cases had pulmonary sequestration. Conclusions CCAMis a rare disease which can be discovered along with pulmonary infection.Multiple gas cysts are the most common imaging findings and the preoperative diagnosis of CCAM is mostly based on chest CT examination. Type Ⅰ and type Ⅱ are the most common pathological classification.The surgical resection should be given early surgi-cal resection and the prognosis is usually good.
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Asthma is the most common chronic respiratory disease in children. Currently,inhaled corti-costeroids is the most effective drug for asthma,however,there are some adverse effect by long-time using. Ac-cording to recent literature,long-time treatment with inhaled corticosteroids may inhibit height growth. But this suppression only occurs within early stage of asthma treatment, and have no effect or slight effect on adult height. The adverse effects of inhaled corticosteroids for height are also related to its iypes,dose and usage.
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The non-structural (NS1) protein is a multifunctional molecular protein encoded by influenza A virus genome. NS1 plays an important role in inhibition of host immune responses. In order to assess the cellular localization of NS1 in different influenza A virus subtypes, we performed the immunofluorescence assay to observe the cellular location of NS1 after infection with influenza A virus WSN (H1N1), PR8 (H1N1), CA04 (H1N1), SD (H9N2) and AH01 (H7N9) in A549 cells and MDCK cells respectively. According to the results, NS1-WSN and NS1-PR8 accumulated mainly in cytoplasm at 24 h post infection, while the NS1-CA04 and NS1-SD appeared major in the nucleus. We also observed localization of NS1 by transfected 293T cells with plasmids which encoding the full-length NS1 from WSN, SD and AH01. The key sites which might determine the different cellular localization of NS1 were chosen by sequence alignment, and seven residues which were different between WSN, PR8 and CA04, SD and AH01 were finally focused. However, we found that single mutation of these residues could not alter the localization of NS1. The data indicated that the difference of location might not be caused by substitution of a single site, which contributes to our understanding of the diverse regulation of host factors during different subtypes of influenza virus infection.
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Raman spectroscopy has generated many branches during the development for more than 90 years. Surface enhanced Raman spectroscopy (SERS) improves SNR by using the interaction between tested materials and the surface of rough metal, as to quickly get higher sensitivity and precision spectroscopy without sample pretreatment. This article describes the characteristic and classification of SERS, and updates the theory and clinical application of SERS. It also summarizes the present status and progress of SERS in various disciplines and illustrates the necessity and urgency of its research, which provides rationale for the application for SERS in microbiology.