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M13-IN5重组噬菌体的构建与鉴定及其对沙眼衣原体的作用研究 被引量:4
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作者 练婷婷 魏世娟 +6 位作者 刘原君 任杰 王生 郭媛丽 郭睿 刘全忠 邵丽丽 《中华皮肤科杂志》 CAS CSCD 北大核心 2018年第12期859-864,共6页
目的 构建针对沙眼衣原体的活性噬菌体,并评估其对沙眼衣原体的作用.方法 将衣原体噬菌体phiCPG1衣壳蛋白VP1的IN5序列与M13噬菌体重组,获得重组M13-IN5噬菌体.利用PCR扩增、酶切、测序重组噬菌体基因,验证目的片段是否成功插入;通过噬... 目的 构建针对沙眼衣原体的活性噬菌体,并评估其对沙眼衣原体的作用.方法 将衣原体噬菌体phiCPG1衣壳蛋白VP1的IN5序列与M13噬菌体重组,获得重组M13-IN5噬菌体.利用PCR扩增、酶切、测序重组噬菌体基因,验证目的片段是否成功插入;通过噬菌斑形成实验检测重组噬菌体的活性.利用CCK8法检测效价为1011噬斑形成单位(PFU)/ml的M13噬菌体和M13-IN5重组噬菌体对Hela细胞增殖的影响,同时设置空白对照组(未感染衣原体的Hela细胞).Western印迹检测重组噬菌体、M13噬菌体、对数期大肠杆菌ER2738中IN5环蛋白的表达.在沙眼衣原体E型标准株的培养过程中,分别加入效价为1011PFU/ml的M13噬菌体、M13-IN5重组噬菌体进行干预,并设置衣原体对照组.感染后36 h,共聚焦显微镜下观察M13噬菌体和M13-IN5重组噬菌体定位情况;在感染后36、48、60、72 h碘染色观察计数包涵体.两样本均数间比较用t检验;多组样本均数间比较采用方差分析,两两比较采用Bonferroni检验.结果 成功构建含有IN5环基因的具有生物活性的重组M13噬菌体,Western印迹证实重组噬菌体表达IN5环/pⅢ融合蛋白,重组噬菌体效价达3.05×1011 PFU/ml.CCK8法检测显示,空白对照组、M13噬菌体组、M13-IN5噬菌体组A450值分别为3.63±0.01、3.55±0.02、3.70±0.01,组间差异无统计学意义(F=12.0,P>0.05).共聚焦显微镜定位显示,噬菌体荧光与衣原体包涵体荧光存在重叠;衣原体感染后36、72h,M13-IN5噬菌体组和M13噬菌体组包涵体数目较对照组减少(均P< 0.05),且M13-IN5噬菌体组包涵体数目较M13噬菌体组有明显降低(P<0.05);衣原体感染后48、60h,M13噬菌体组、M13-IN5噬菌体组与对照组包涵体数目差异无统计学意义(P>0.05).结论 构建的M13-IN5重组噬菌体具有生物学活性,且能够成功表达IN5环蛋白;体外实验中,该噬菌体能进入衣原体包涵体内,且能明显抑制沙眼衣原体感染. 展开更多
关键词 沙眼衣原体 细菌噬菌体M13 DNA 重组 包涵体 in5
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M13-IN5重组噬菌体对沙眼衣原体的抑制机制
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作者 王江懿 尤聪 +5 位作者 赵乐然 王梅 魏世娟 练婷婷 邵丽丽 刘全忠 《中国皮肤性病学杂志》 CAS CSCD 北大核心 2022年第8期877-883,共7页
目的探索重组M13-IN5噬菌体抑制沙眼衣原体(Chlamydia trachomatis,Ct)生长作用的机制。方法通过噬菌斑法测定M13-IN5噬菌体的滴度;CCK-8法检测M13-IN5噬菌体和青霉素对HeLa细胞的毒性作用;将不同滴度的M13-IN5噬菌体与50 U/mL的青霉素... 目的探索重组M13-IN5噬菌体抑制沙眼衣原体(Chlamydia trachomatis,Ct)生长作用的机制。方法通过噬菌斑法测定M13-IN5噬菌体的滴度;CCK-8法检测M13-IN5噬菌体和青霉素对HeLa细胞的毒性作用;将不同滴度的M13-IN5噬菌体与50 U/mL的青霉素作用于沙眼衣原体后,通过免疫荧光计数包涵体数量;将滴度为10^(7)pfu/mL M13-IN5噬菌体与50 U/mL的青霉素作用于沙眼衣原体后进行子代感染力实验;采用定量逆转录PCR检测两组干预后CT_046、CT_111、CT_395、CT_443、CT_666、CT_823基因的转录变化,未干预沙眼衣原体感染组为阳性对照。两样本均数间比较采用t检验,多组样本均数间比较采用单因素ANOVA方法,多个样本间两两比较采用Bonferroni校正。结果滴度≤10^(9)pfu/mL的M13-IN5噬菌体以及50 U/mL的青霉素对HeLa细胞无毒性作用,10^(7)pfu/mL M13-IN5噬菌体与50 U/mL青霉素干预后,48 h形成的包涵体数量无明显区别,抑制率分别为19.62%和21.09%(P>0.05);M13-IN5噬菌体组干预形成的子代包涵体数量较青霉素干预组减少20.26%(P<0.05)。CT_046、CT_443在两个干预组转录水平均下降,M13-IN5噬菌体组较青霉素组下降明显(P<0.05);M13-IN5噬菌体干预组CT_666转录水平下降(P<0.05),青霉素干预组无变化(P>0.05);青霉素干预组压力应答基因CT_111、CT_395、CT_823转录水平增高(P<0.05),M13-IN5噬菌体干预组以上基因转录水平无变化(P>0.05)。结论重组M13-IN5噬菌体对沙眼衣原体的抑制作用强于青霉素,CT_046、CT_443、CT_666基因是重组噬菌体抑制沙眼衣原体生长的作用靶点。 展开更多
关键词 沙眼衣原体 M13噬菌体 in5 青霉素
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Targeting c-Myc on cell growth and vascular endothelial growth factor expression in IN500 glioblastoma cells 被引量:2
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作者 HU Yu-hua KONG Shi-qi +2 位作者 KONG Hai-bo WU Jian-liang CHEN Ze 《Chinese Medical Journal》 SCIE CAS CSCD 2012年第11期2025-2031,共7页
Background The level of c-Myc is closely associated with high pathological grade and the poor prognosis of gliomas. Vascular endothelial growth factor (VEGF) is the most important angiogenic factor that potently sti... Background The level of c-Myc is closely associated with high pathological grade and the poor prognosis of gliomas. Vascular endothelial growth factor (VEGF) is the most important angiogenic factor that potently stimulates the proliferation and migration of vascular endothelial cells. This study aimed to address the biological importance of c-Myc in the development of gliomas, we downregulated the expression of c-Myc in the human glioblastoma cell line IN500 and studied the in vitro effect on cellular growth, proliferation, and apoptosis and the expression of VEGF and the in vivo effect on tumor formation in a xenograft mouse model. Methods IN500A cells were stably transfected with shRNA-expressing plasmids for either c-Myc (pCMYC-shRNA) or as a control (pCtrl-shRNA). Following establishment of stable cells, the mRNA expressions of c-Myc and VEGF were examined by reverse transcription (RT)-PCR, and c-Myc and VEGF proteins by Western blotting and immunohistochemistry. Cell-cycle progression and apoptosis were determined by flow cytometry. The in vivo effect of targeting c-Myc was determined by subcutaneous injection of stable cells into immunodeficient nude mice. Results The stable transfection of pCMYC-shRNA successfully knocked down the steady-state mRNA and protein levels of c-Myc in IN500, which positively correlated with the downregulation of VEGF. Downregulating c-Myc in vitro also led to G1-S arrest and enhanced apoptosis. In vivo, targeting c-Myc reduced xenograft tumor formation and resulted in significantly smaller tumors. Conclusions c-Myc has multiple functions in glioblastoma development that include regulating cell-cycle, apoptosis, and VEGF expression. Targeting c-Myc expression may be a promising therapy for malignant glioma. 展开更多
关键词 C-MYC RNA interference in5OOA cells cell proliferation apoptosis vascular endothelial growth factor GLIOMA
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金属/三元乙丙橡胶粘接工艺研究 被引量:10
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作者 邹德荣 王维明 《粘接》 CAS 2002年第1期43-44,52,共3页
研究了IN5 0 1胶粘剂的配比 ,烘干、抽真空工艺和M— 2 5表面处理剂对金属 /EPDM粘接性能的影响 ,对确定的最佳技术方法进行了重现性和加速老化试验 ,结果表明 ,IN5 0 1胶粘剂A/B的质量配比为 5∶3时 ,对金属表面用M— 2 5处理 ,涂刷的... 研究了IN5 0 1胶粘剂的配比 ,烘干、抽真空工艺和M— 2 5表面处理剂对金属 /EPDM粘接性能的影响 ,对确定的最佳技术方法进行了重现性和加速老化试验 ,结果表明 ,IN5 0 1胶粘剂A/B的质量配比为 5∶3时 ,对金属表面用M— 2 5处理 ,涂刷的胶粘剂的试件抽真空 15min 。 展开更多
关键词 in5d胶粘剂 粘接工艺 三元乙丙橡胶 金属
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纳米针状In(OH)_3中残留氯元素的存在状态研究
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作者 陈霞 李晨辉 《理化检验(物理分册)》 CAS 2007年第10期514-516,共3页
采用高分辨透射电子显微术(HRTEM)分析纳米针状In(OH)3粉末中氯元素的存在状态。分析结果显示,氯元素主要以In5Cl9的形式存在,并且未探测到其他种类的InxCly化合物。In5Cl9中最容易观察到的晶面为(110)面(d=0.617 nm)和(113)面(d=0.428 ... 采用高分辨透射电子显微术(HRTEM)分析纳米针状In(OH)3粉末中氯元素的存在状态。分析结果显示,氯元素主要以In5Cl9的形式存在,并且未探测到其他种类的InxCly化合物。In5Cl9中最容易观察到的晶面为(110)面(d=0.617 nm)和(113)面(d=0.428 nm)。 展开更多
关键词 in5Cl9 In(OH)3 高分辨透射电子显微术
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一种新的用于删除选择标记基因的Cre/lox系统 被引量:2
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作者 袁媛 刘允军 王涛 《Acta Botanica Sinica》 CSCD 2004年第7期862-866,共5页
设计了一种新的诱导型Cre/lox似系统,并在转基因烟草(Mcofiana tabacum L.)中进行了验证。在诱导剂的作用下,位于同向lox位点之间的选择标记基因(hpt)和重组酶基因(Cre)在烟草愈伤组织中被删除。在该系统中,Cre基因在玉米乙酰苯胺类... 设计了一种新的诱导型Cre/lox似系统,并在转基因烟草(Mcofiana tabacum L.)中进行了验证。在诱导剂的作用下,位于同向lox位点之间的选择标记基因(hpt)和重组酶基因(Cre)在烟草愈伤组织中被删除。在该系统中,Cre基因在玉米乙酰苯胺类化合物诱导启动:(In5-2)的控制下表达。对转基因后代的分子检测结果表明,不论是否加入了诱导剂,目的基因(gus)均被整合到烟草基因组中;在诱导剂处理的48株转基因烟草T0代中,45株的hpt基因被删除了。该系统只使用一个载体,克服了二次转化系统带来的问题。 展开更多
关键词 乙酰苯胺类化合物诱导启动子(in5-2) CRE/LOX 无标记 转基因烟草
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5E康复模式在血液透析初期沟通中的应用 被引量:9
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作者 温玉 谢翠芳 +5 位作者 罗梅 张蓉 李文秀 王燕 谢祝君 曾春兰 《求医问药(下半月刊)》 2012年第6期464-466,共3页
目的:为让终末期肾病血液透析初期患者从未经血液净化的明显尿毒症状态能平稳的过渡到日常透析阶段,为了满足这一特殊人群的需求,使这一转折点能平稳过渡,我科在血液透析初期患者的医患沟通中运用5E康复模式,保证治疗、护理工作正常进... 目的:为让终末期肾病血液透析初期患者从未经血液净化的明显尿毒症状态能平稳的过渡到日常透析阶段,为了满足这一特殊人群的需求,使这一转折点能平稳过渡,我科在血液透析初期患者的医患沟通中运用5E康复模式,保证治疗、护理工作正常进行。方法:选择自2001年以来在我院进行血液透析的终末期肾病患者460例,随机分为观察组及对照组,观察组按5E康复模式为患者制定透析初期教育路径表,并对患者及家属进行宣教指导。对照组则是用传统方法,即在透析前、透析中、透析后随机进行教育指导。结果:观察组以减少康复的延迟及资源的浪费,提高患者的透析质量、自理能力及医从性,取得较满意的效果。结论:5E康复模式教育对提高血液透析患者生存质量和延长存活时间具有重要的意义。 展开更多
关键词 终末期肾病 血液透析 尿毒症 医患沟通 5E康复模式
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五年制师范体育教学中培养女生创新能力的研究
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作者 王超 钱晓年 《湖北体育科技》 2013年第9期830-831,共2页
体育教师应从分析女生对体育课认识单一、固化的原因着手,从女生生理、心理特点出发,在体育教学内容、方法、实践中渗透,探讨提高师范女生体育创新能力的途径与方法,对女生积极参加体育锻炼和养成健身习惯起到正面强化作用,从而为师范... 体育教师应从分析女生对体育课认识单一、固化的原因着手,从女生生理、心理特点出发,在体育教学内容、方法、实践中渗透,探讨提高师范女生体育创新能力的途径与方法,对女生积极参加体育锻炼和养成健身习惯起到正面强化作用,从而为师范女生走上讲台打下良好的身体素质基础。 展开更多
关键词 五年制师范女生 体育教学 创新能力 途径与方法
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Relationship between the Brittlement Susceptibility and H Permeation Current for UNS G11180 Steel in 5% NaCl Solution with H_2S 被引量:1
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作者 Xueyuan ZHANG and Yuanlong DU(State Key Lab. for Corrosion and Protection, Institute of Corrosion and Protection of Metals,Chinese Academy of Sciences, Shenyang 110015, China) 《Journal of Materials Science & Technology》 SCIE EI CAS CSCD 1998年第6期555-558,共4页
Sulfide stress corrosion cracking (SSCC) behaviour of UNS G11180 steel in 5% NaCl solution with H2S was studied by slow strain rate tensile test (SSRT), SEM and electrochemical hydro gen permeation technique. The resu... Sulfide stress corrosion cracking (SSCC) behaviour of UNS G11180 steel in 5% NaCl solution with H2S was studied by slow strain rate tensile test (SSRT), SEM and electrochemical hydro gen permeation technique. The results reveal different cracking mechanism and H permeation current (IH) through UNS G11180 steel plate in different concentration of H2S solution. The susceptibility to SSCC of UNS G11180 Steel in 5% NaCl solution with H2S was evaluated by the permeation current(IH, μA), which depends on the concentration (c×10-6) of H2S by the equation:IH = 8.525 ×c0.7249. lt is proved that the electrochemical H permeation method is a practical way to assess the susceptibility to SSCC. 展开更多
关键词 UNS NaCl Solution with H2S Relationship between the Brittlement Susceptibility and H Permeation Current for UNS G11180 Steel in 5
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Changes in 5α-reductase (type 2) gene expression in epididymis of puberty diabetic rats
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作者 Zhong-Shan WANG Hong-Jun WANG Su-Huan LIU Dept.of Cell Biology and Dept.of Molecular Biology,Jilin University,Changchun 130021,China 《Asian Journal of Andrology》 SCIE CAS CSCD 2001年第2期157-157,共1页
The changes in 5α-reductase (type 2 ) gene expression in the epi-didymis of puberty diabetic rats were studied by the Northern blot and Dotblot method. Rats were divided into 3 groups: the control group (C), thediabe... The changes in 5α-reductase (type 2 ) gene expression in the epi-didymis of puberty diabetic rats were studied by the Northern blot and Dotblot method. Rats were divided into 3 groups: the control group (C), thediabetic group (D), and the diabetic group with insulin treatment (DI).Results: The Northern blot intensity of the caput epididymis in Group D is 展开更多
关键词 gene expression in epididymis of puberty diabetic rats type 2 Changes in 5 REDUCTASE
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MULTI-HOP RELAYING IN 5G: FROM RESEARCH TO SYSTEMS, STANDARDS, AND APPLICATIONS
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作者 lei guo keping long +1 位作者 kumudu munasinghe xuetao wei 《China Communications》 SCIE CSCD 2016年第10期I0001-I0002,共2页
In recent years,the demands of high traffic transmission motivate the rapid development of wireless access techniques,and it becomes promising to design the fifth generation(5G)wireless networks.Essential requirements... In recent years,the demands of high traffic transmission motivate the rapid development of wireless access techniques,and it becomes promising to design the fifth generation(5G)wireless networks.Essential requirements for 5G involve higher traffic volume,indoor or hotspot traffic,and spectrum,energy,and cost efficien- 展开更多
关键词 MULTI-HOP RELAYING IN 5G STANDARDS AND APPLICATIONS FROM RESEARCH TO SYSTEMS
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Phase Transition and Quasinormal Modes for Spherical Black Holes in 5D Gauss–Bonnet Gravity
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作者 Ming Zhang Rui-Hong Yue 《Chinese Physics Letters》 SCIE CAS CSCD 2018年第4期7-11,共5页
We study the quasinormal modes(QNMs) of massless scalar perturbations to probe the van der Waals like SBH/LBH phase transition of anti-de Sitter black holes in five-dimensional(5D) Gauss–Bonnet gravity. It is fou... We study the quasinormal modes(QNMs) of massless scalar perturbations to probe the van der Waals like SBH/LBH phase transition of anti-de Sitter black holes in five-dimensional(5D) Gauss–Bonnet gravity. It is found that the signature of this SBH/LBH phase transition is detected when the slopes of the QNMs frequency change drastically and differently in small and large black holes near the critical point. The obtained results further support that the QNMs can be a dynamic probe to investigate the thermodynamic properties in black holes. 展开更多
关键词 der Phase Transition and Quasinormal Modes for Spherical Black Holes in 5D Gauss Bonnet Gravity
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Developing high-Q × f value MgNb_(2–x)Ta_(x)O_(6)(0≤x≤0.8) columbite ceramics and clarifying the impact mechanism of dielectric loss: Crystal structure, Raman vibrations, microstructure, lattice defects, chemical bond characteristics, structural parameters, and microwave dielectric properties in-depth studies 被引量:1
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作者 Lingxia Li Yuting Li +1 位作者 Jianli Qiao Mingkun Du 《Journal of Materials Science & Technology》 SCIE EI CAS CSCD 2023年第15期186-199,共14页
With the increasing frequency of wireless mobile communication,dielectric materials with higher Q×f are required to enhance the signal strength of microwave passive devices and thus compensate for the rapid atten... With the increasing frequency of wireless mobile communication,dielectric materials with higher Q×f are required to enhance the signal strength of microwave passive devices and thus compensate for the rapid attenuation of high-frequency signals during propagation.In order to improve the Q×f of the MgNb2O6 system and study the internal mechanism between the structure and microwave dielectric properties,Ta5+ion is introduced to design and fabricate MgNb_(2-x)Ta_(x)O_(6)(0≤x≤0.8)ceramics by the solid-state reaction method.The single columbite phase and dense structure are observed throughout the entire composition range.All elements(Mg,Nb,Ta,and O)can be detected by X-ray photoelectron spectroscopy.Insights from the Raman spectroscopy and group theoretical analysis reveal that the Ag(2)and Ag(3)modes located at∼905 cm−1 dominate the Raman vibration.In addition,theɛr depends on the polarizability reflected by the C–M equation and Raman shift.Theτf closely relates to the bond energy of the Nb/Ta–O bond and restoring force characterized by average octahedral distortion of[Nb/TaO6].Notably,the crucial factors for the huge improvement of Q×f are clarified deeply,in terms of the lower internal strain,the reduced ordering-induced domain size,and the chemical bond valence closer to the ionic valence state,which promotes the stability of crystal structure.Moreover,the introduction of lower electronegative ions enhances the ability of electrons to compensate for oxygen vacancies,thereby reducing the dielectric loss caused by the conductance.Excellent microwave properties(ɛr=19.2,Q×f=170,000 GHz,andτf=−55.6 ppm℃–1)are achieved in MgNb1.4Ta0.6O6 ceramic,specifically a nearly 40%increase in the Q×f.This work has enriched the theory associated with low dielectric loss and provided the MgNb_(1.4)Ta_(0.6)O_(6) ceramic with a pretty high-Q×f for the application in the design of the 5G/6G wireless communication devices. 展开更多
关键词 Advances in 5G/6G materials High-Q×f value ceramic Group theoretical analysis Oxygen vacancy Chemical bond characteristics Microwave dielectric properties
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Nearly 1 in 5 U. S. Counties Have Unhealthy Air 被引量:1
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作者 Chris Baltimore 岳俊 姜淑萱 《当代外语研究》 2004年第6期13-15,共3页
[选译者言]臭氧(ozone),化学名O3,具有特殊鱼腥味的蓝色气体,具强氧化性,在一定浓度下即可转化成氧气,没有任何残留,不会形成二次污染,被誉为最清洁的氧化剂和消毒剂。位于地球大气层的臭氧层,可吸收各种有害的宇宙射线,保护人类不受伤... [选译者言]臭氧(ozone),化学名O3,具有特殊鱼腥味的蓝色气体,具强氧化性,在一定浓度下即可转化成氧气,没有任何残留,不会形成二次污染,被誉为最清洁的氧化剂和消毒剂。位于地球大气层的臭氧层,可吸收各种有害的宇宙射线,保护人类不受伤害。 自然界中的活氧含量约为0.04PPM,可防止病菌的异常增长,保护生态的自然平衡。臭氧对空气中大肠杆菌、金黄色葡萄球菌等的杀灭率达99%以上,臭氧的灭菌速度要比氯快300到600倍,比紫外线快3000倍。 但是,凡事有利即有弊,据医学专家介绍,如果臭氧在空气中的含量超标,会对人体健康有极大影响。科学研究表明,人体暴露在高浓度臭氧环境中,会出现皮肤刺痒、眼睛刺痛、呼吸不畅、咳嗽和头痛等症状。如果暴露时间较长,就会导致短暂性肺功能异常,引起肺部组织受伤。有过敏体质的人,如果长时间暴露在臭氧含量超过每立方米0.18毫克的环境里,可能会导致慢性肺病,甚至产生肺纤维化等永久伤害。本文介绍了美国在对臭氧含量立法工作上的最新进展。 展开更多
关键词 环保署 Counties Have Unhealthy Air Nearly 1 in 5 U
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