鼻咽癌组织中miR-10b、LMP1和Twist1的表达变化及临床意义

目的 观察鼻咽癌组织中微小RNA(miR)-10b、潜伏期膜蛋白1(LMP1)及Twist同系物1(Twist1)的表达变化,并探讨其临床意义.方法 采用实时定量PCR法检测92例鼻咽癌患者癌组织及癌旁组织中的miR-10b、LMP1和Twist1 mRNA.采用免疫印迹法检测癌组织及癌旁组织中的LMP1、Twist1...     

EB病毒潜伏膜蛋白1致癌机制的研究进展

EB病毒（Epstein-Barrvirus，EBV）是一种人类r-疱疹病毒，也是最早被识别的人类肿瘤病毒，它与许多淋巴增生性疾病如传染性单核细胞增多症以及恶性肿瘤如伯基特淋巴瘤、何杰金氏淋巴瘤、鼻咽瘤等的病病有关。它是在全球广泛分布的双链DNA病毒，EBV感染人群后通常在人体内终身潜伏，亦可导致感染的宿主细胞不断发生恶性转化，形成肿瘤。已证实EBV与Burking淋巴瘤（BL）、肺癌、胃癌和鼻咽癌（nasopharyngeal carcinoma，NPC）有关。随着研究的不断深入，人们发现EBV不仅与上述肿瘤的发生相关，同时也与潜伏感染基因有关，其中潜伏膜蛋白1（1atent membrane protein1，LMP1）并诱导B细胞转化，如纤维母细胞系癌基因转化。LMP1在肿瘤的发生、发展中起主要的作用，目前被证实有促进细胞转化甚至恶变的性质。但是LMP1是唯一被发现的EBV相关的B细胞恶性肿瘤产生的EBV编码蛋白，     

鼻咽癌组织中潜伏膜蛋白的表达变化及意义

EB病毒编码的潜伏膜蛋白（LMP1）是目前惟一被证实具有转化上皮细胞功能的基因产物。2006年10月-2008年9月,我们检测了鼻咽癌组织中的LMP1,并探讨其与鼻咽癌发生发展的关系。现报告如下。     

鼻咽癌组织中FHIT和LMP-1表达及意义

目的探讨脆性组氨酸三联体基因（FHTT）和潜伏膜蛋白1（LMP-1）在鼻咽癌发生、发展中的作用。方法采用免疫组化两步法检测63份鼻咽癌组织（鼻咽癌组）和30份正常鼻咽黏膜组织（对照组）FHIT和LMP—1表达,分析其与鼻咽癌临床病理特征的相关性。结果鼻咽癌组FHIT的阳性率显著低于对照组（p〈0．01）,LNP-1阳性率显著高于对照组（P〈0．01）;FHIT和LMP-1阳性率与鼻咽癌病理分级、临床分期和淋巴结转移密切相关（P均〈0．05）;鼻咽癌中FHIT与LMP-1表达呈负相关（r=-0．421,P〈0．05）。结论FHIT低表达、IMP-1高表达可促进鼻咽癌的发生、发展。     

EB病毒潜伏膜蛋白LMP1疫苗的预测及验证

目的 采用生物信息学方法分析EB病毒潜伏膜蛋白LMP1,为疫苗的研发提供理论依据。方法 在NCBI数据库中获取LMP1蛋白的基因组序列和氨基酸序列,采用生物信息学分析工具ProtParam、SOPMA、SWISS MODEL、SignalP、TMHMM、Cell-PLoc 2.0、NetNGly、NetPhos-3.1、Conserved domains、IEDB、BLAST、Immunomedicine Group、UniProt预测LMP1蛋白的理化性质,二、三级结构,信号肽和跨膜区域,亚细胞定位,糖基化位点和磷酸化位点,保守结构域,B、T细胞表位,同源性,抗原决定簇以及相互作用蛋白。构建重组质粒LMP1-pMV261以及重组卡介苗,并通过检测LMP1分子及蛋白表达水平探究构建疫苗的基本条件。结果 LMP1蛋白分子式为C₁₉₀₁H₂₈₇₇N₄₉₃O₅₆₂S₁₁,氨基酸数为386,相对分子质量为41.98238×10³,原子总数为5 844。LMP1蛋白...     

EBV潜伏感染膜蛋白1在肺癌组织中的表达及意义

目的探讨潜伏感染膜蛋白1（LMP1）在肺组织中的表达意义。方法采用免疫组化法对手术切除的肺癌组织（108例）及癌旁正常肺组织（22例）中的EB病毒（EBV）潜伏感染膜蛋白1（LMP1）进行检测，并用图像分析法进行形态学定量。结果肺癌及癌旁组织中的LMP1阳性率分别为6．5％和0，两者比较无统计学差异（P〉0．05），但癌组织中LMP1的平均面积明显高于癌旁组织（P〈0．05），表明癌组织中的LMP1表达量高于癌旁组织。结论LMP1在肺组织细胞的恶性转化中可能起一定作用。     

EB病毒血清学指标筛查鼻咽癌的研究进展

鼻咽癌在我国华南地区高发,早诊早治可提高鼻咽癌患者的生存率,因此提高该地区人群早期鼻咽癌检出率十分关键。EB病毒(Epstein-Barr virus,EBV)感染鼻咽上皮细胞是导致鼻咽癌发生的重要因素,EBV多种基因分别表达R反式激活因子(R transactivator,Rta)、Z反式激活因子(Z transactivator,Zta)、早期抗原(early antigen,EA)和病毒衣壳抗原(viral capsid antigen,VCA)以及EB病毒核抗原1(EBV nuclear antigen 1,EBNA1)等多种蛋白,由此在人体血清中产生以免疫球蛋白G(immunoglobulin G,IgG)和免疫球蛋白A(immunoglobulin A,IgA)为主的抗体,其中Rta-IgG、Zta-IgG/IgA、EA-IgA、VCA-IgA、EBNA1-IgA/IgG为鼻咽癌的特异标志物。该文综述相关基因及其表达蛋白的生物学特性,以及应用这些血清学标志物筛查鼻咽癌高发区人群的诊断价值,为开展早期筛查鼻咽癌研究提供依据。     

壮族鼻咽癌患者EB病毒抗体与年龄的关系

目的探讨广西地区壮族患者EB病毒各年龄段EBNA1/IgA、EA/IgG及EA/IgA抗体的rA值和阳性率与年龄的关系。方法收集140例未经治疗的壮族鼻咽癌患者和280例健康人的血清,用ELISA检测EBNA1/IgA、EA/IgG及EA/IgA抗体,分别计算各年龄段的抗体水平及阳性率并进行统计学分析。结果鼻咽癌患者各年龄段EBNA1/IgA、EA/IgA及EA/IgG抗体rA值均有统计学差异(P<0.05)。在50岁以后的年龄段病人与健康人EA/IgG、EA/IgA、EBNA1/IgA抗体阳性率无显著差异。结论壮族鼻咽癌患者EB病毒EBNA1/IgA、EA/IgA及EA/IgG抗体水平存在年龄上的差异。在鼻咽癌的临床诊断和高危人群筛查中有必要根据不同年龄段的人群界定不同的阳性临界值。     

鼻型结外NK/T细胞淋巴瘤患者血清潜伏膜蛋白1/EB病毒核抗原1检测的临床意义

目的 检测鼻型结外NK/T细胞淋巴瘤(ENKL)患者血清EB病毒核抗原1(EBNA1)、潜伏膜蛋白1(LMP1),探讨其与ENKL的关系.方法 选取首都医科大学北京同仁医院2010年8月到2013年8月新诊断ENKL患者36例、健康对照者20例,以EBNA1、LMP1为目的基因,采用实时定量PCR方法分别检测ENKL患者治疗前后和健康对照者外周血EBNA1、LMP1.结果 治疗前,ENKL患者血清EBNA1中位数为1.9×104(0 ～ 11.0×104)拷贝/μl,健康对照组为8.0(0 ～43.8)拷贝/μl;ENKL患者LMP1中位数为3.9 ×103(118.3 ～24.0×103)拷贝/μl,健康对照组为3.3(0～33.3)拷贝/μl.治疗前ENKL患者EBNA1、LMP1均显著高于健康对照组(P均＜0.01).治疗后,ENKL患者血清EBNA1中位数为1.0×103(0 ～2.0×103)拷贝/μl,LMP1为300.8(0 ～825.7)拷贝/μl,两者较治疗前均显著降低(P＜0.05).治疗有效患者的治疗后EBNA1、LMP1中位数均低于治疗无效的患者(P＜0.05).治疗前ENKL患者EBNA1、LMP1与血清乳酸脱氢酶水平呈正相关(r=0.364、0.546,P=0.040、0.012).EBNA1、LMP1低于临界值(EBNA1:1.3×104拷贝/μl; LMP1:3.0×103拷贝/μl)患者的2年总生存率和无进展生存率优于EBNA1、LMP1高于临界值者,但差异无统计学意义(P均＞0.05).结论 (1)检测ENKL患者血清EBNA1和LMP1对评判治疗效果具有一定价值;(2)ENKL患者血清EBNA1、LMP1可反映肿瘤负荷.     

Therapeutic LMP1 polyepitope vaccine for EBV-associated Hodgkin disease and nasopharyngeal carcinoma

Development of an epitope-based vaccination strategy designed to enhance Epstein-Barr virus (EBV)-specific CD8(+) cytotoxic T lymphocytes (CTLs) is increasingly being considered as a preferred approach for the treatment of EBV-associated relapsed Hodgkin disease (HD) and nasopharyngeal carcinoma (NPC). EBV-encoded latent membrane proteins, LMP1 and LMP2, are the only target antigens available for therapeutic augmentation of CTL responses in patients with HD and NPC. Here, we describe preclinical studies using a recombinant poxvirus vaccine that encodes a polyepitope protein comprising 6 HLA A2-restricted epitopes derived from LMP1. Human cells infected with this recombinant polyepitope construct were efficiently recognized by LMP1-specific CTL lines from HLA A2 healthy individuals. Furthermore, immunization of HLA A2/K(b) mice with this polyepitope vaccine consistently generated strong LMP1-specific CTL responses to 5 of the 6 epitopes, which were readily detected by both ex vivo and in vitro assays. More important, this polyepitope vaccine successfully reversed the outgrowth of LMP1-expressing tumors in HLA A2/K(b) mice. These studies provide an important platform for the development of an LMP-based polyepitope vaccine as an immunotherapeutic tool for the treatment of EBV-associated HD and NPC.     

细胞角蛋白-19基因在甲状腺癌中的表达及意义

目的探讨细胞角蛋白-19（CK-19）mRNA在甲状腺癌中的表达及其意义。方法用RT—PCR方法检测甲状腺癌（27例）、良性甲状腺疾病（18例）及正常甲状腺（9例）组织中CK-19mRNA的表达。结果CK-19mRNA在甲状腺癌组中表达为2．63±2．14,与非癌组相比．P〈0．05;在有淋巴转移的甲状腺癌组的表达为3．41±2．63,明显高于无转移者的1．79±0．97,P〈0．05。结论CK-19mRNA在甲状腺癌组织中表达明显增强,且与淋巴结转移有关,提示CK-19可能参与了甲状腺癌的发生发展,并对肿瘤转移有促进作用。     

MicroRNA-19b promotes macrophage cholesterol accumulation and aortic atherosclerosis by targeting ATP-binding cassette transporter A1

Rationale

Macrophage accumulation of cholesterol leads to foam cell formation which is a major pathological event of atherosclerosis. Recent studies have shown that microRNA (miR)-19b might play an important role in cholesterol metabolism and atherosclerotic diseases. Here, we have identified miR-19b binding to the 3′UTR of ATP-binding cassette transporter A1 (ABCA1) transporters, and further determined the potential roles of this novel interaction in atherogenesis.

Objective

To investigate the molecular mechanisms involved in a miR-19b promotion of macrophage cholesterol accumulation and the development of aortic atherosclerosis.

Methods and results

We performed bioinformatics analysis using online websites, and found that miR-19b was highly conserved during evolution and directly bound to ABCA1 mRNA with very low binding free energy. Luciferase reporter assay confirmed that miR-19b bound to 3110-3116 sites within ABCA1 3′UTR. MiR-19b directly regulated the expression levels of endogenous ABCA1 in foam cells derived from human THP-1 macrophages and mouse peritoneal macrophages (MPMs) as determined by qRT-PCR and western blot. Cholesterol transport assays revealed that miR-19b dramatically suppressed apolipoprotein AI-mediated ABCA1-dependent cholesterol efflux, resulting in the increased levels of total cholesterol (TC), free cholesterol (FC) and cholesterol ester (CE) as revealed by HPLC. The excretion of ³H-cholesterol originating from cholesterol-laden MPMs into feces was decreased in mice overexpressing miR-19b. Finally, we evaluated the proatherosclerotic role of miR-19b in apolipoprotein E deficient (apoE^−/−) mice. Treatment with miR-19b precursor reduced plasma high-density lipoprotein (HDL) levels, but increased plasma low-density lipoprotein (LDL) levels. Consistently, miR-19b precursor treatment increased aortic plaque size and lipid content, but reduced collagen content and ABCA1 expression. In contrast, treatment with the inhibitory miR-19b antisense oligonucleotides (ASO) prevented or reversed these effects.

Conclusion

MiR-19b promotes macrophage cholesterol accumulation, foam cell formation and aortic atherosclerotic development by targeting ABCA1.     

Interferon-α2b Treatment for COVID-19 Is Associated with Improvements in Lung Abnormalities

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection causes coronavirus disease 2019 (COVID-19), a lung disease that may progress to systemic organ involvement and in some cases, death. The identification of the earliest predictors of progressive lung disease would allow for therapeutic intervention in those cases. In an earlier clinical study, individuals with moderate COVID-19 were treated with either arbidol (ARB) or inhaled interferon (IFN)-α2b +/−ARB. IFN treatment resulted in accelerated viral clearance from the upper airways and in a reduction in the circulating levels of the inflammatory biomarkers IL-6 and C-reactive protein (CRP). We have extended the analysis of this study cohort to determine whether IFN treatment had a direct effect on virus-induced lung abnormalities and also to ascertain whether any clinical or immune parameters are associated with worsening of lung abnormalities. Evidence is provided that IFN-α2b treatment limits the development of lung abnormalities associated with COVID-19, as assessed by CT images. Clinical predictors associated with worsening of lung abnormalities include low CD8+ T cell numbers, low levels of circulating albumin, high numbers of platelets, and higher levels of circulating interleukin (IL)-10, IL-6, and C-reactive protein (CRP). Notably, in this study cohort, IFN treatment resulted in a higher percentage of CD8+ T cells, lower tumor necrosis factor (TNF)-α levels and, as reported earlier, lower IL-6 levels. Independent of treatment, age and circulating levels of albumin and CRP emerged as the strongest predictors of the severity of lung abnormalities.     

Lung Transplant Recipients Immunogenicity after Heterologous ChAdOx1 nCoV-19—BNT162b2 mRNA Vaccination

(1) Background: High immunosuppressive regimen in lung transplant recipients (LTRs) hampers the immune response to vaccination. We prospectively investigated the immunogenicity of heterologous ChAdOx1 nCoV-19-BNT162b2 mRNA vaccination in an LTR cohort. (2) Methods: Forty-nine COVID-19 naïve LTRs received a two-dose regimen ChAdOx1 nCoV-19 vaccine. A subset of 32 patients received a booster dose of BNT162b2 mRNA vaccine 18 weeks after the second dose. (3) Results: Two-doses of ChAdOx1 nCoV-19 induced poor immunogenicity with 7.2% seropositivity at day 180 and low neutralizing capacities. The BNT162b2 mRNA vaccine induced significant increases in IgG titers with means of 197.8 binding antibody units per milliliter (BAU/mL) (95% CI 0–491.4) and neutralizing antibodies, with means of 76.6 AU/mL (95% CI 0–159.6). At day 238, 32.2% of LTRs seroconverted after the booster dose. Seroneutralization capacities against Delta and Omicron variants were found in only 13 and 9 LTRs, respectively. Mycophenolate mofetil and high-dose corticosteroids were associated with a weak serological response. (4) Conclusions: The immunogenicity of a two-dose ChAdOx1 nCoV-19 vaccine regimen was very poor in LTRs, but was significantly enhanced after the booster dose in one-third of LTRs. In immunocompromised individuals, the administration of a fourth dose may be considered to increase the immune response against SARS-CoV-2.     

鼻咽癌组织中AKT2的表达及意义

目的 探讨AKT2在鼻咽癌组织中的表达变化及意义. 方法 采用免疫组化EliVision二步法检测50例鼻咽癌组织和25例慢性鼻咽炎组织中的AKT2,并分析其与鼻咽癌临床病理参数的关系. 结果 AKT2在鼻咽癌组织中的阳性表达率为48.0%(24/50),与慢性鼻咽炎组织的8.0%(2/25)相比,P<0.01;AKT2的表达与鼻咽癌淋巴结转移及临床分期相关(P均<0.05). 结论 AKT2在鼻咽癌的发生、发展中可能发挥重要作用,可作为预测鼻咽癌发生及侵袭转移的参考指标之一.