变应性鼻炎模型小鼠鼻黏膜的形态学观察

目的:建立小鼠变应性鼻炎模型,观察小鼠鼻腔黏膜组织的重塑情况。方法:20只BALB/c小鼠被随机分为致敏组和对照组,使用卵清蛋白(OVA)诱导建立小鼠变应性鼻炎模型。通过HE染色观察小鼠鼻黏膜的大体重塑情况,吉姆萨染色观察嗜酸性粒细胞,阿辛蓝-过碘酸-希夫染色观察杯状细胞;酶联免疫吸附(ELISA)法检测小鼠血清中白细胞介素-4(IL-4)的水平。结果:小鼠变应性鼻炎模型的生物学行为评分为6.5±1.3,提示造模成功。与对照组相比,致敏组鼻腔黏膜出现上皮细胞脱落、坏死,杯状细胞增生,鳞状上皮化生,固有层和黏膜下层腺体增生、血管扩张,组织水肿,固有层内可见特征性的嗜酸性粒细胞浸润,造模后鼻腔黏膜结构存在重塑。致敏组小鼠鼻黏膜嗜酸性粒细胞计数及杯状细胞计数分别为(26.4±5.72)和(24.14±3.12),而对照组分别是(8.31±2.42)和(9.41±1.22),两组比较均具有统计学差异(P0.05);致敏组血清中白细胞介素4(IL-4)水平为(18.9±3.1)pg/ml,对照组为(8.3±1.4)pg/ml,致敏组显著高于对照组,差异有统计学意义(P0.05)。结论:通过卵清蛋白诱导建立的小鼠变应性鼻炎模型鼻腔黏膜存在组织重塑。     

Toll样受体2及Toll样受体4在大鼠实验性变应性鼻炎中的表达

目的研究Toll样受体2（Toll-like receptor2,TLR2）及Toll样受体4（TLR4）在实验性变应性鼻炎大鼠鼻黏膜中的表达及脂多糖（Lipopolysaccharide,LPS）对其表达的影响。方法SD大鼠48只随机分为正常对照组（A组）;变应性鼻炎组（B组）：经腹腔注射及鼻腔滴入卵清白蛋白（Ovalbumin,OVA）建立变应性鼻炎（Allergic rhinitis,AR）模型;变应性鼻炎＋LPS刺激组（C组）：大鼠激发成变应性鼻炎模型后再以LPS滴鼻。用逆转录聚合酶链反应（RT-PCR）方法检测鼻黏膜中TLR2 mRNA、TLR4 mRNA的表达。结果B、C组大鼠均成功激发为AR动物模型;各组鼻黏膜中均有TLR2 mRNA、TLR4 mRNA表达;各组间TLR2 mRNA的表达差异无统计学意义（P〉0．05）。B、C组TLR4 mRNA的表达较A组高（P〈0．01）;C组TLR4 mRNA表达较B组增高（P〈0．01）。结论AR大鼠有TLLR4的表达增高;LPS刺激后TLR4表达进一步增高,说明TLR4可能参与AR的发病。TLR2在AR大鼠中的表达未见增高;LPS刺激后。TLR2表达未见进一步增高,TLR2与变应性鼻炎的关系有待进一步研究。     

抗IL-6抗体治疗小鼠变应性鼻炎的作用的研究

目的：探讨IL-6 抗体治疗小鼠变应性鼻炎的作用。方法：实验动物分三组：PBS 组（正常对照组）、抗IL-6 抗体组（以抗IL-6 单 克隆抗体处理）、IgG 抗体对照组（以IgG对照抗体处理）。应用卵清蛋白（OVA）致敏建立小鼠变应性鼻炎模型,给予抗IL-6 单克 隆抗体干预,计量抗原激发后小鼠搔鼻数量。应用HE染色方法检测对小鼠鼻黏膜炎症影响,应用ELISA 法检测小鼠灌洗液中 IL-4、IFN-r含量。结果：治疗后抗IL-6 抗体组小鼠搔鼻症状相对于抗体对照组明显减轻（P<0.01)。与PBS 组比较,IgG 抗体对照组 小鼠鼻腔灌洗液中总炎性细胞数,淋巴细胞数及嗜酸性粒细胞数明显升高（P<0.05);IL-6 抗体治疗后,小鼠鼻腔灌洗液中总炎性 细胞数,淋巴细胞数及嗜酸性粒细胞数明显降低（P<0.05);IgG抗体对照组小鼠鼻腔灌洗液IL-4 含量明显升高（P<0.01),而IFN-r 含量不变（P>0.05);IL-6 抗体治疗后,IL-4 含量较IgG 抗体对照组降低（P<0.05),IFN-r含量不变（P>0.05)。结论：IL-6 抗体可有效 治疗小鼠变应性鼻炎。     

甲苯-2,4-二异氰酸酯经鼻致敏建立羊变应性鼻炎模型

目的旨在建立鼻腔宽敞的大型动物变应性鼻炎模型,初步探讨其实用性。方法①南江黄羊4只行鼻部解剖,记录鼻腔解剖学参数。②南江黄羊12只,8只为模型组,15%甲苯-2,4-二异氰酸酯（TDI）橄榄油溶液滴鼻致敏,4只为对照组,使用橄榄油液。记录建模过程中黄羊症状体征评分,建模完成后测定黄羊鼻腔灌洗液组胺含量并行鼻黏膜组织病理学检查。③将建模成功的黄羊随机分为A组（布地奈德治疗组）和B组（生理盐水对照组）,记录治疗前后症状体征评分变化,评价该模型对药物治疗的反应。结果①黄羊鼻腔宽敞,鼻腔解剖结构与人类极其相似。②TDI致敏后,与对照组相比,模型组8只黄羊均出现典型变应性鼻炎症状体征,鼻腔灌洗液中组胺含量明显增高,差异均有统计学意义（P〈0.01）;组织病理学检查见黄羊鼻黏膜下组织水肿,血管扩张,固有层内散在或灶性以嗜酸性粒细胞为主的炎症细胞浸润。③布地奈德治疗组症状体征评分下降,与对照组相比差异有统计学意义（P〈0.05）。结论成功建立大型动物变应性鼻炎模型,不但可用于研究药物疗效,还可用于判定新的物理和手术治疗安全性及有效性。     

Calpeptin通过抑制GATA3减轻变应性鼻炎大鼠炎症

摘要 目的：本研究旨在评估钙蛋白酶抑制剂calpeptin减轻变应性鼻炎大鼠炎症的作用并探讨其机制。方法：将20只雄性SD大鼠采用数字表法随机分为4组：正常组（Normal）、变应性鼻炎组（AR）、地塞米松干预AR组（DXMS+AR）、calpeptin干预AR组（Calpeptin+AR）。造模成功后,对大鼠AR症状进行行为学评分,对大鼠鼻黏膜组织切片以HE和PAS染色法观察鼻黏膜病理改变;对大鼠外周血以ELISA法检测总IgE、IL-4、IL-13水平;对大鼠鼻黏膜组织以免疫蛋白印迹法检测GATA3蛋白表达水平。单因素方差分析进行多组间比较,LSD- t检验进行组间两两比较。结果：与Normal组相比,AR组大鼠的鼻部过敏症状、鼻黏膜嗜酸粒细胞计数及外周血总IgE水平均升高,Calpeptin与地塞米松均能减轻气道炎症,减少嗜酸性粒细胞浸润,降低血清中OVA诱导的IgE的生成。探讨机制发现,酶联免疫吸附试验检测Th2细胞因子,与Normal组比较,AR组血清IL-4、IL-13水平均升高（P＜0.05）,而Calpeptin与地塞米松均能降低血清IL-4、IL-13水平（P＜0.05）。免疫蛋白印迹法检测大鼠鼻黏膜GATA3蛋白表达水平显示,与Normal组比较,AR组鼻黏膜 GATA3表达升高（P＜0.05）,而Calpeptin与地塞米松组鼻黏膜 GATA3表达均下降（P＜0.05）。结论：腹腔注射calpeptin能够减轻变应性鼻炎大鼠局部和全身过敏反应,其机制可能下调GATA3表达,影响Th2细胞的分化及细胞因子的分泌有关。     

SB239063通过抑制TNF-α和p38MAPK，减轻香烟烟雾暴露变应性鼻炎大鼠MKP-1的表达

摘要 目的：探讨p38MAPK抑制剂SB239063对香烟烟雾暴露变应性鼻炎大鼠p38MAPK信号通路、TNF-α、MKP-1表达的影响。方法：先构建变应性鼻炎（AR）大鼠模型,然后给予AR大鼠被动吸入香烟烟雾（CS）,再行腹腔注射AR大鼠SB239063（100 mg/kg）,分为AR组、AR+CS组、AR+CS+SB239063组。造模后对各组大鼠进行症状学评分;苏木精-伊红（HE）染色法观察大鼠鼻黏膜的形态学变化;实时荧光定量PCR（RT-PCR）检测鼻黏膜p38MAPK、MKP-1 mRNA的表达水平;酶联免疫吸附试验(ELISA)分析外周血、脾脏TNF-α的含量;Western blots检测鼻黏膜p38MAPK、p-p38MAPK、MKP-1蛋白的表达水平。结果：与AR组比较,AR+CS组大鼠的过敏症状（P<0.05）加重;鼻黏膜嗜酸性粒细胞（P<0.05）、中性粒细胞浸润（P<0.01）增多; MKP-1 mRNA及其蛋白的表达水平均升高（P<0.001）;外周血、脾脏TNF-α的表达水平升高（P<0.001）;p-p38MAPK蛋白的表达水平升高（P<0.001）。与AR+CS组比较,AR+CS+SB239063组大鼠过敏症状评分下降（P<0.01）;鼻黏膜中嗜酸性粒细胞、中性粒细胞计数下降（P值均< 0.05）;p38MAPK mRNA的表达水平降低（P<0.05）,MKP-1 mRNA及其蛋白的表达水平下降（P<0.001）;外周血、脾脏TNF-α的表达水平降低（P值均< 0.001）;p38MAPK、p-p38MAPK蛋白的表达水平降低,差异均具有显著统计学意义（P值均 < 0.001）。结论：SB239063通过抑制TNF-α、p38MAPK及其自磷酸化,减轻香烟烟雾暴露变应性鼻炎大鼠MKP-1的表达。     

儿童过敏性结膜炎与变应性鼻炎的相关性及鼻眼联合防治效果分析

目的：探讨儿童过敏性结膜炎与变应性鼻炎的相关性研究及鼻眼联合防治的临床效果。方法：回顾性分析300 例儿童过敏性 结膜炎与310 例儿童变应性鼻炎患者的临床资料,对儿童过敏性结膜炎与变应性鼻炎的相关性进行分析后将所有患儿随机均分 为对照组与观察组,对照组采用常规点眼的方法进行治疗,观察组则采用鼻朗喷鼻联合人工泪液点眼进行治疗。比较两组临床疗 效及不良反应情况。结果：（1）300 例过敏性结膜炎患儿中,50 例（16.67%）并发变应性鼻炎;310 例变应性鼻炎患儿中,59 例 （19.03%）并发过敏性结膜炎（P＞0.05）;（2）109 例同时并发两种疾病患儿中,均进行眼结膜与鼻粘膜的刮片检查嗜酸性粒细胞, 其中60 例（55.05%）结膜刮片与67 例（61.47%）鼻粘膜刮片检测到嗜酸性粒细胞（P＞0.05）;（3）两组治疗前后BUT 及角膜荧光素 染色评分、症状评分、临床总有效率比较差异明显（P＜0.05）。结论：儿童过敏性结膜炎与变应性鼻炎具有一定的相关性;鼻朗喷鼻 联合人工泪液点眼治疗儿童合并变应性鼻炎的临床疗效显著。     

顺尔宁对变应性鼻炎患者血清肺表面活性蛋白A、D动态水平影响及近 期疗效观察

目的:探讨顺尔宁对变应性鼻炎患者体内肺表面活性蛋白A、D水平的影响及近期疗效观察。方法:纳入的变应性鼻炎患者分成单纯变应性鼻炎患者组不伴哮喘组106例,变应性鼻炎伴哮喘患者组75例,健康成人组20例作为实验对照组。连续应用顺尔宁8周后,统计患者鼻部总体症状评分,以及血清SP-A、SP-D水平的动态变化情况。结果:单纯变应性鼻炎组、变应性鼻炎伴哮喘组患者治疗4周后、8周后与治疗前比较,鼻部总体症状评分均明显降低,差异均有统计学意义(均P0.05)。治疗4周后,单纯变应性鼻炎组、变应性鼻炎伴哮喘组患者血清SP-A、D水平较治疗前,差异无统计学意义(均P0.05);治疗8周后,单纯变应性鼻炎组、变应性鼻炎伴哮喘组患者血清SP-A、D水平较治疗前明显下降(均P0.05)。结论:顺尔宁可影响变应性鼻炎患者体内SP-A、SP-D水平,改善鼻部不适的症状。     

趋化因子CCL-18 在慢性鼻- 鼻窦炎中的表达及意义

目的:分析趋化因子CCL-18在不同组织病理特征慢性鼻-鼻窦炎和正常鼻黏膜的表达差异,探讨CCL-18在慢性鼻-鼻窦炎中的表达及意义。方法:采用苏木精-伊红染色(HE),Masson染色及过碘酸-雪夫(PAS)染色对慢性鼻-鼻窦炎组织进行病理分析。采用Western blot检测CCL-18蛋白水平在不同组织病理特征慢性鼻-鼻窦炎和正常鼻黏膜组织中的表达差异。结果:CCL-18蛋白水平在伴鼻息肉和不伴有鼻息肉慢性鼻窦炎均较正常鼻黏膜组织中显著上调(P<0.05)。CCL-18蛋白水平在嗜酸性粒细胞慢性鼻窦炎的表达水平明显高于非嗜酸性粒细胞慢性鼻窦炎(P<0.05)。腺体型,纤维炎症型及水肿型慢性鼻-鼻窦炎中CCL-18表达水平均高于正常鼻黏膜,以水肿型表达最为显著(P<0.05)。结论:CCL-18在嗜酸性粒细胞和水肿型慢性鼻窦炎中高度表达,提示CCL-18可能参与慢性鼻-鼻窦炎中嗜酸性粒细胞的浸润这一基本病理过程。     

白细胞介素-33在慢性鼻-鼻窦炎伴鼻息肉患者黏膜组织中的表达及意义

目的观察慢性鼻-鼻窦炎伴鼻息肉(CRSwNP)患者鼻黏膜组织中白细胞介素-33(IL-33)的表达与嗜酸性粒细胞(Eos)的浸润情况,探讨IL-33与Eos在其发病中作用及相关性。方法采用免疫组织化学LSAB法分别检测CRSwNP患者(观察组)32例和单纯鼻中隔偏曲患者(对照组)24例的鼻黏膜组织中IL-33的表达情况;苏木精-伊红染色下观察炎性细胞浸润程度并对Eos计数;全血细胞分析法测定两组患者血液中嗜酸性粒细胞含量。结果 (1)CRSwNP组织中上皮层及间质内Eos、中心粒细胞为主的炎性细胞及部分腺体内有IL-33的表达;(2)免疫组织化学检测显示IL-33阳性细胞数、苏木精-伊红染色Eos计数均高于对照组,差异具有统计学意义(P0.05)。Eos计数与IL-33阳性细胞数在CRSwNP中存在正相关性(P0.05);(3)血液中Eos含量无统计学意义。结论 IL-33是一种新型的细胞因子,在CRSwNP患者体内高表达,可能与嗜酸性粒细胞浸润互相促进,在CRSwNP发生发展中具有重要的作用。     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.