Immunogenicity and protective efficacy of adenoviral and subunit RSV vaccines based on stabilized prefusion F protein in pre-clinical models

Respiratory Syncytial Virus (RSV) remains a leading cause of severe respiratory disease for which no licensed vaccine is available. We have previously described the derivation of an RSV Fusion protein (F) stabilized in its prefusion conformation (preF) as vaccine immunogen and demonstrated superior immunogenicity in naive mice of preF versus wild type RSV F protein, both as protein and when expressed from an Ad26 vaccine vector. Here we address the question if there are qualitative differences between the two vaccine platforms for induction of protective immunity. In naïve mice, both Ad26.RSV.preF and preF protein induced humoral responses, whereas cellular responses were only elicited by Ad26.RSV.preF. In RSV pre-exposed mice, a single dose of either vaccine induced cellular responses and strong humoral responses. Ad26-induced RSV-specific cellular immune responses were detected systemically and locally in the lungs. Both vaccines showed protective efficacy in the cotton rat model, but Ad26.RSV.preF conferred protection at lower virus neutralizing titers in comparison to RSV preF protein. Factors that may contribute to the protective capacity of Ad26.RSV.preF elicited immunity are the induced IgG2a antibodies that are able to engage Fcγ receptors mediating Antibody Dependent Cellular Cytotoxicity (ADCC), and the induction of systemic and lung resident RSV specific CD8 + T cells. These data demonstrate qualitative improvement of immune responses elicited by an adenoviral vector based vaccine encoding the RSV preF antigen compared to the subunit vaccine in small animal models which may inform RSV vaccine development.     

程序性死亡受体1和程序性死亡受体配体1在非小细胞肺癌组织中的表达情况及临床意义

目的探讨程序性死亡受体1(PD-1)和程序性死亡受体配体1(PD-L1)在非小细胞肺癌(NSCLC)组织中的表达情况及临床意义。方法选择150例NSCLC患者的NSCLC组织及其癌旁组织,采用实时荧光定量聚合酶链反应(PCR)检测两种组织中PD-1 mRNA和PD-L1 mRNA的相对表达量。采用免疫组织化学染色法检测NSCLC组织中PD-1和PD-L1的表达情况,分析PD-1和PD-L1表达情况与患者临床特征的关系。采用流式细胞术检测NSCLC组织和癌旁组织中CD4^+-PD-1、CD8^+-PD-1、CD14^+-PD-L1、CD68^+-PD-L1的表达水平。结果NSCLC组织中PD-1 mRNA和PD-L1 mRNA的相对表达量分别为(5.03±1.92)和(4.95±1.09),分别高于癌旁组织的(1.72±0.81)和(1.25±0.24),差异均有统计学意义(P﹤0.05)。TNM分期为Ⅲ~Ⅳ期、低分化、有淋巴结转移、有远处转移的NSCLC患者NSCLC组织中PD-1和PD-L1的高表达率均明显高于TNM分期为Ⅰ~Ⅱ期、高+中分化、无淋巴结转移、无远处转移的患者,差异均有统计学意义(P﹤0.01)。NSCLC组织中CD4^+-PD-1、CD8^+-PD-1、CD14^+-PD-L1、CD68^+-PD-L1的表达水平均明显高于癌旁组织,差异均有统计学意义(P﹤0.01)。结论PD-1和PD-L1在NSCLC组织中高表达,可能成为一种新的生物标志物,PD-1/PD-L1信号通路可能参与了NSCLC的免疫逃逸过程,对其逃逸机制进行研究可以为NSCLC患者的临床治疗提供新靶点。     

Therapeutic efficacy of intense pulsed light in patients with refractory meibomian gland dysfunction

Purpose

To evaluate the efficacy and safety of intense pulsed light (IPL) combined with meibomian gland expression (MGX) for treatment of refractory meibomian gland dysfunction (MGD).

Aberrant expression of Wnt/β-catenin signaling pathway genes in aggressive malignant gastric gastrointestinal stromal tumors

IntroductionRecent reports on gene expression profiling (GEP) show several genes associated with malignant progression of GIST. However, genes associated with malignant transformation have not been clarified. Here, we aimed to reveal distinct genes in aggressive malignant GIST, using comprehensive gene expression analysis.Materials and methodsWe investigated GEP obtained by microarrays for 43 gastric GISTs, which mostly harbored KIT and PDGFRA mutations and integrated clinicopathological risk information. RT-PCR and immunohistochemistry were performed for FZD7, a receptor of Wnt ligands.ResultsGEP divided 43 gastric GISTs into two clusters. A cluster included seven of eight high-risk GISTs (88%) in modified NIH classification and was defined as high-risk cluster; the other cluster was defined as low-risk cluster. The number of probes with over 3-fold changes between the two clusters was 1,177, in which probes corresponding to 16 oncogenes were included. Genes involved in the Wnt signaling pathway were the most abundant among the 16 oncogenes. Focusing on 73 Wnt signaling pathway genes of the 21,578 probes, 12 upregulated and 5 downregulated genes were found in the high-risk cluster. Major cascade genes promoting the Wnt/β-catenin signaling pathway, including WNT11, FZD family, and DVL2, were upregulated in the high-risk cluster. SNAI1, SNAI2, and BIRC5, which are activated by this pathway and increase cell proliferation, were also upregulated. These gene expression alterations were consistent in the positive direction of this pathway. GISTs in high-risk cluster strongly expressed FZD7.ConclusionWnt/β-catenin signaling pathway may play an important role in malignant transformation of indolent GIST.     

乙酰肝素酶、E-cadherin、N-cadherin和vimentin蛋白在胃癌中的表达及其意义

目的： 探讨人胃癌组织中乙酰肝素酶（HPA）、上皮标志物E-cadherin、间质标志物N-cadherin和vimentin蛋白的表达及其与人胃癌临床病理指标的关系，以及HPA与E-cadherin、N-cadherin、vimentin蛋白之间的关系。方法： 应用免疫组织化学方法检测91例人胃癌组织中HPA、E-cadherin、N-cadherin和vimentin蛋白表达情况；采用卡方检验，检测这几种蛋白阳性表达率与人胃癌不同病理指标的关系；同时采用Spearman等级相关分析HPA与E-cadherin、N-cadherin、vimentin蛋白之间的关系。结果： 人胃癌组织中HPA、E-cadherin、N-cadherin和vimentin蛋白阳性表达率为75.82%、51.65%、54.95%和23.08%。91例人胃癌组织中HPA、E-cadherin、N-cadherin和vimentin蛋白的阳性表达率与胃癌患者年龄、性别、胃癌大小均无明显相关（P > 0.05），而与人胃癌发生部位、分化程度和有无淋巴结转移、侵袭程度有关（P < 0.05）。HPA、N-cadherin和vimentin蛋白在贲门部阳性表达率明显高于胃体、幽门及胃窦胃癌；低分化者、有淋巴结转移者和胃癌侵袭深处这3种蛋白阳性表达率明显高于高分化者、无淋巴结转移者和胃癌起始部位；E-cadherin蛋白在幽门及胃窦部胃癌阳性表达率明显高于贲门和胃体胃癌；高中分化者、无淋巴结转移者明显高于低分化者和有淋巴结转移者。E-cadherin蛋白阳性表达率在侵袭深处明显低于起始部位。经Spearman等级相关分析显示，人胃癌组织中HPA与E-cadherin蛋白的阳性表达率呈负相关（r=-0.341，P < 0.05）；而与N-cadherin（r=0.366，P < 0.05）和vimentin（r=0.284，P < 0.05）蛋白阳性表达率呈正相关。结论： 人胃癌组织中HPA、N-cadherin和vimentin蛋白在低分化者、有淋巴结转移者和胃癌侵袭深处高表达；E-cadherin蛋白在高中分化胃癌，无淋巴结转移者和起始部位高表达。HPA与N-cadherin、vimentin蛋白阳性表达率呈正相关，HPA与E-cadherin蛋白的阳性表达率呈负相关，可见HPA蛋白与间质标志物N-cadherin和vimentin蛋白表达增强有关，而与上皮标志物E-cadherin蛋白缺失或减少有关，因此HPA可能诱导人胃癌组织发生上皮间质转化。     

Is the pepsin immunohistochemical staining of laryngeal lesions an available way for diagnosing laryngopharyngeal reflux

Abstract

Backgound: Pepsin immunohistochemical (IHC) staining is a promising diagnostic approach of laryngopharyngeal reflux (LPR). The interarytenoid mucosa has been proved to be an effective biopsy area.

Objectives: To investigate whether positive result of pepsin IHC staining in laryngeal lesions can predict LPR.

Methods: The study included 136 patients with laryngeal cancer or vocal cord leukoplakia. 24?h multichannel intraluminal impedance-pH (MII-pH) was performed before operation, and pepsin IHC staining was performed on pathological sections after operation. The results of the two methods were compared.

Results: Among the 136 patients, 101 with at least one LPR event were regarded as MII-pH positive group, and another 35 were negative. The positive rate of pepsin IHC staining was 93.1% in MII-pH positive group and 54.1% in MII-pH negative group (p?<?.05). If the MII-pH results were used as a reference, the sensitivity and specificity of pepsin IHC staining in the diagnosis of LPR were 93.1 and 45.7%, respectively. The consistency of pepsin IHC and MII-pH was moderate (Kappa value = 0.452).

Conclusions: The sensitivity of pepsin IHC staining in laryngeal lesions for diagnosing LPR is satisfactory. The existence of false negative of MII-pH may be the main reason for the low specificity.     

天冬氨酸及赖氨酸2～10肽单链和双链的合成与毒性

目的探讨同种氨基酸形成多肽的毒性和应用前景。方法用固相合成法合成天冬氨酸和赖氨酸的2~10肽单链与双链化合物,通过小鼠尾静脉将这些化合物注射到小鼠肝脏内,观察它们对小鼠的致死作用(LD50)。结果当天冬氨酸单链2~10肽的浓度依次达到0.15、0.1、0.09、0.03、0.0065、0.03、0.034、0.035和0.04 mol/L时,小鼠的死亡率达到50%。当赖氨酸单链2~10肽的浓度依次达到0.28、0.1、0.047、0.0225、0.0028、0.00166、0.0015、0.0011和0.00075 mol/L时,小鼠的死亡率达到50%。当天冬氨酸-赖氨酸双链2~10肽的浓度依次得到0.5、0.05、0.017、0.014、0.009、0.006、0.004、0.004、0.004、0.004 mol/L时,小鼠的死亡率达到50%。结论单链多肽的毒性大于双链多肽。在2~6肽范围内,天冬氨酸单链多肽随着氨基酸数目增加毒性逐渐增加;相反,在7~10肽范围内,随着氨基酸数目的增加毒性逐渐降低。在2~10肽范围内,赖氨酸单链多肽随着氨基酸数目增加毒性逐渐增加。在2~6肽范围内,天冬氨酸-赖氨酸双链多肽随着氨基酸数目增加毒性逐渐增加;但在7~10肽范围内,其毒性相似。