可注射原位交联海藻酸钙骨修复材料小鼠体内异位成骨研究

目的对复合了人重组骨形态发生蛋白2(rhBMP-2)的可注射原位交联海藻酸钙骨修复材料在小鼠体内异位成骨进行评估。方法实验组为含0．1mg rhBMP-2的材料0．1ml,空白对照组为不含rhBMP-2 的材料0．1ml,分别注射到小鼠左后肢大腿肌陷窝中,对照组植入含0．1mg rh BMP-2的骨优导,21天后, 影像学检查成骨情况,解剖称骨湿重,同时在注射后7天,14天,21天解剖实验组动物各三只,取注射部位进行HE染色组织学检查。结果 21天影像学检查显示实验组小鼠左后肢肌陷窝处有骨痂生成,解剖取异位成骨称湿重实验组为239．2±59．7mg,对照组为225．5±56．9mg,无显著性差异,2周后组织学检查有大量骨髓细胞及骨小梁生成。结论该材料有良好的诱导新骨生成的能力。     

自发性甲状腺皱缩结节与甲状腺乳头状癌的超声鉴别诊断研究

目的探究自发性甲状腺皱缩结节常规超声和超声造影检查下(contrast-enhanced ultrasound,CEUS)的表现特点,并与甲状腺乳头状癌(papillary thyroid carcinomas,PTC)进行鉴别诊断。方法回顾性分析2019年2月至2020年1月于南昌大学第一附属医院就诊且经细针穿刺(fine-needle aspiration,FNA)细胞学或术后病理证实的39例自发性甲状腺皱缩结节患者(46个结节,即皱缩结节组),其中男12例,女27例,年龄25~76 岁,以及同时期经术后病理确诊的PTC患者中随机选取32例(36个癌灶,即PTC组),其中男8例,女24例,年龄23~68 岁。比较2组病变的大小、形状、边界、回声、钙化、血流信号等常规超声表现,其中28个皱缩结节及30个PTC在FNA或术前行CEUS检查,分析比较2组在增强模式下的表现。采用两独立样本t检验、χ2检验或Fisher精确检验比较2组的超声表现,差异有统计学意义者行二元Logistic回归检验是否为独立预测风险因素。结果单变量分析示皱缩结节组与PTC组相比,更常表现纵横比<1、边界清晰、极低回声以及无或点线状增强方式,差异有统计学意义(纵横比<1者∶36比17,χ2 =8.511;边界清晰:30比15,χ2 =4.523;极低回声:27比9,χ2=9.310;无或点线状增强方式:24比3,χ2 =33.369;P值均<0.05),多变量Logistic回归分析示结节纵横比<1、边界清晰及极低回声为皱缩结节的独立风险预测因子(OR值分别为5.204、3.134和5.042,P值分别为0.003、0.031和0.003)。其中15个皱缩结节从良性表现至首次可疑恶性的时间跨度为(18.6±10.5)个月,初始最大长径缩小(24.3±11.4)mm。结论皱缩结节相较于PTC超声检查中更常表现为纵横比<1、边界清晰、极低回声和无或点线状增强方式。结合随访过程中患者历史超声检查有助于诊断。     

不完全分隔内耳畸形患者人工耳蜗植入研究进展

不完全分隔内耳畸形作为内耳畸形的一种,是导致重度、极重度感音神经性聋的病因之一.其曾被认为是人工耳蜗植入手术的禁忌症.但是随着相关研究的进展,人工耳蜗植入已成为其主要治疗手段.本文就不完全分隔内耳畸形的概念、分类、及其所致的重度、极重度感音神经性聋患者人工耳蜗植入手术的相关研究进展作一综述.     

T cell receptor β‐chain repertoire analysis reveals intratumour heterogeneity of tumour‐infiltrating lymphocytes in oesophageal squamous cell carcinoma

Oesophageal squamous cell carcinoma (ESCC) has a generally poor prognosis, due to the lack of effective treatment methods. Immunotherapeutic approaches based on tumour‐infiltrating lymphocytes (TILs) have demonstrated that durable responses are produced in some patients with solid tumours, which suggests the potential feasibility of clinical application of immunotherapy for ESCC. However, many of the basic characteristics of TILs in ESCC are poorly understood, including clonality, specificity and spatial heterogeneity of the response of TILs, which depends on the interaction between antigens and T cell receptors (TCRs). We used ultra‐deep sequencing of rearranged genes in TCR β‐chain (TCRβ) to profile the basic characteristics of T cells in tumour tissues (four to six regions from each tumour) as well as matched adjacent normal tissue and peripheral blood from seven patients diagnosed with primary ESCC. We found that T cell clones within ESCCs were quite different from those of the peripheral blood and even the adjacent normal tissues in general. Although there was a relatively higher degree of overlap of intratumoural TCRβ repertoires than those between the tumour and other tissues, intratumoural TCRβ repertoires were spatially heterogeneous. Due to the restricted sampling, high‐throughput TCRβ sequencing could characterize the diversity and composition of a limited (compartment‐dependent) fraction of the respective T cell clones in any individual ESCC, expanding our understanding of immune behaviour and immune response and shedding more light on ESCC immunotherapy. Copyright © 2016 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

Plasmacytoid dendritic cells and type 1 interferon promote peripheral expansion of forkhead box protein 3+ regulatory T cells specific for the ubiquitous RNA‐binding nuclear antigen La/Sjögren's syndrome (SS)‐B

RNA‐binding nuclear antigens are a major class of self‐antigen to which immune tolerance is lost in rheumatic diseases. Serological tolerance to one such antigen, La/Sjögren's syndrome (SS)‐B (La), is controlled by CD4⁺ T cells. This study investigated peripheral tolerance to human La (hLa) by tracking the fate of hLa‐specific CD4⁺ T cells expressing the transgenic (Tg) 3B5.8 T cell receptor (TCR) after adoptive transfer into lymphocyte‐replete recipient mice expressing hLa as a neo‐self‐antigen. After initial antigen‐specific cell division, hLa‐specific donor CD4⁺ T cells expressed forkhead box protein 3 (FoxP3). Donor cells retrieved from hLa Tg recipients displayed impaired proliferation and secreted interleukin (IL)?10 in vitro in response to antigenic stimulation. Transfer of highly purified FoxP3‐negative donor cells demonstrated that accumulation of hLa‐specific regulatory T cells (T_reg) was due primarily to expansion of small numbers of donor T_reg. Depletion of recipient plasmacytoid dendritic cells (pDC), but not B cells, severely hampered the accumulation of FoxP3⁺ donor T_reg in hLa Tg recipients. Recipient pDC expressed tolerogenic markers and higher levels of co‐stimulatory and co‐inhibitory molecules than B cells. Adoptive transfer of hLa peptide‐loaded pDC into mice lacking expression of hLa recapitulated the accumulation of hLa‐specific T_reg. Blockade of the type 1 interferon (IFN) receptor in hLa Tg recipients of hLa‐specific T cells impaired FoxP3⁺ donor T cell accumulation. Therefore, peripheral expansion of T_reg specific for an RNA‐binding nuclear antigen is mediated by antigen‐presenting pDC in a type 1 IFN‐dependent manner. These results reveal a regulatory function of pDC in controlling autoreactivity to RNA‐binding nuclear antigens.     

TGF-beta(1) gene modified immature dendritic cells exhibit enhanced tolerogenicity but induce allograft fibrosis in vivo

Administration of donor-derived immature dendritic cells (DC) can prolong the survival of MHC-mismatched cardiac allografts. Genetic modification of DC by immunosuppressive molecules can enhance their potential tolerogenicity. In this study bone marrow derived immature DC were genetically modified by transforming growth factor (TGF) beta1 by recombinant Ad. TGF-beta(1) gene modified immature DC (TGF-beta-DC) displayed a characteristic phenotype of immature DC, decreased ability to secrete interleukin 12, and reduced allostimulatory ability. TGF-beta-DC induced alloantigen-specific T cell hyporesponsiveness in vitro and in vivo, and Th2 cytokine polarization. mRNA expression of donor MHC class II (Ia(b)) and human TGF-beta(1) was detected in spleen and lymph nodes of the allogeneic recipients for 3 weeks after TGF-beta-DC infusion, indicating that microchimerism of TGF-beta-DC is exhibited in allogeneic recipients. In this murine cervical heterotopic heart transplantation model, the survival of the allograft in recipients intravenously infused with TGF-beta-DC 7 days before transplantation was greatly prolonged, and about 67% of cardiac grafts survived more than 40 days. Histological analysis of the allografts showed that the normal myocardial architecture was well preserved, accompanied by very little necrotic cells, but interstitial fibrosis replaced myocytes, and moderate collagen suffused the whole cardiac allograft in the recipients infused with TGF-beta-DC. mRNA expression of type III procollagen was markedly increased in the allografts of the recipients infused with TGF-beta-DC. Our results suggest that infusion of TGF-beta(1) gene modified immature DC prolongs the survival of the allograft through the effective induction of donor-specific T cell hyporesponsiveness. However, TGF-beta(1) expressed by gene modified immature DC can cause the fibrosis of the allografts, which may limit the application of this approach in the allograft transplantation.     

Adenovirus-mediated intratumoral lymphotactin gene transfer potentiates the antibody-targeted superantigen therapy of cancer

Bacterial superantigens are extremely potent activators of murine and human T lymphocytes. To engineer superantigens for cancer immunotherapy, staphylococcal enterotoxin A (SEA) was genetically fused to the Fab region of the human colon carcinoma-reactive monoclonal antibody (mAb) C215. Fusion protein C215Fab-SEA can trigger cytotoxic T cells against C215 antigen positive tumor cells and induce tumor-suppressive cytokines. However, the antitumor effect of C215Fab-SEA is often not satisfactory because of T cell deletion after activation and failure to induce potent CTL activity after repeated administration. Lymphotactin (Lptn) is a potent chemoattractant for T cells and NK cells. To improve the therapeutic efficacy of fusion protein C215Fab-SEA we investigated in this study the antitumor responses elicited by combination of C215Fab-SEA and adenovirus-mediated intratumoral Lptn gene transfer in the preestablished C215 antigen expressing B16 melanoma murine model. More significant inhibition of tumor growth and prolonged survival time were observed in tumor-bearing mice that received combined therapy of C215Fab-SEA and Ad-Lptn than those of mice treated with C215Fab-SEA or Ad-Lptn alone. The highest CTL activity of tumor-bearing mice was induced after combined therapy. Intratumoral coadministration of C215Fab-SEA and Ad-Lptn augmented splenic NK activity of tumor-bearing mice most markedly. Our data demonstrate that the in vivo antitumor effect of C215Fab-SEA immunotherapy is potentiated significantly by combination with intratumoral Lptn gene transfer through more efficient induction of specific and nonspecific antitumor immune responses.     

Mitogen activated protein kinase pathway is involved in RhoC GTPase induced motility,invasion and angiogenesis in inflammatory breast cancer

Inflammatory breast cancer (IBC) is the most lethal form of locally advanced breast cancer known. IBC carries a guarded prognosis primarily due to rapid onset of disease, typically within six months, and the propensity of tumor emboli to invade the dermal lymphatics and spread systemically. Although the clinical manifestations of IBC have been well documented, until recently little was known about the genetic mechanisms underlying the disease. In a comprehensive study aimed at identifying the molecular mechanisms responsible for the unique IBC phenotype, our laboratory identified overexpression of RhoC GTPase in over 90% of IBC tumors in contrast to 36% of stage-matched non-IBC tumors. We also demonstrated that overexpression of RhoC GTPase in human mammary epithelial (HME) cells nearly recapitulated the IBC phenotype with regards to invasion, motility and angiogenesis. In the current study we sought to delineate which signaling pathways were responsible for each aspect of the IBC phenotype. Using well-established inhibitors to the mitogen activated protein kinase (MAPK) and phosphatidylinositol-3 kinase (PI3K) pathways. We found that activation of the MAPK pathway was responsible for motility, invasion and production of angiogenic factors. In contrast, growth under anchorage independent conditions was dependent on the PI3K pathway. This revised version was published online in July 2006 with corrections to the Cover Date.     

A novel locus for autosomal dominant nonsyndromic hearing loss identified at 5q31.1-32 in a Chinese pedigree

 Hearing impairment is an extremely heterogeneous disorder. A total of 35 loci and 17 related genes for autosomal dominant nonsyndromic hearing loss have been identified. In a Chinese pedigree characterized by autosomal dominant inheritance with bilateral, postlingual, progressive, and sensorineural nonsyndromic hearing impairment, the putative disease gene locus was localized to chromosome 5q31.1-32 by a genome-wide scan. Fine mapping indicated that the disease gene was located within an 8.8-cM region between markers D5S2056 and D5S638, with a maximum two-point logarithm of differences (LOD) score of 6.89 (θ = 0) at D5S2017. By the candidate gene approach, mutation screening of the DIAPH1 and POU4F3 genes at 5q31 was performed. No mutation was found, suggesting that this is a novel deafness locus, which has been named DFNA42. Received: May 8, 2002 / Accepted: October 1, 2002