Functional Capacity in Adults With Cerebral Palsy: Lower Limb Muscle Strength Matters

Objective

To investigate the relation between lower limb muscle strength, passive muscle properties, and functional capacity outcomes in adults with cerebral palsy (CP).

Dissociation of E-cadherin/β-catenin complex by MG132 and bortezomib enhances CDDP induced cell death in oral cancer SCC-25 cells

E-cadherin/β-catenin complex plays an important role in maintaining the homeostasis of tissues and regulating cell proliferation, survival and apoptosis. To address the relationships between the change of E-cadherin/β-catenin complex and cell apoptosis, human oral squamous carcinoma SCC-25 cells were used to investigate whether the dissociation of the E-cadherin/β-catenin complex was the main reason of MG132- or bortezomib-induced apoptosis. We found that MG132 or bortezomib alone induced remarkable loss of cell integrity and contact, inhibited cell growth, survival, migration and caused cell cycle arrest, intracellular ROS production. Further experiments showed that colony formations were significantly decreased by MG132 and bortezomib alone or plus cis-diaminedichloroplatinum (CDDP). Immunofluorescence staining showed that SCC-25 cells exhibited remarkable accumulations of β-catenin in cytoplasm and few E-cadherin in cell membranes after MG132 or bortezomib treatment. Western blot results showed that MG132 or bortezomib induced high accumulation of ubiquitinated proteins and activation of apoptosis related protein caspase-3. Meanwhile, the combinational use of MG132 or bortezomib with CDDP led to synergistic effects on SCC-25 cells. However, knockdown of β-catenin could decrease MG132 or bortezomib induced cell death. Taken together, our data suggest that the regulation of E-cadherin/β-catenin complex could be a promising therapeutic target to overcome the multidrug resistance of oral cancer.     

骨肉瘤常用化疗药物作用的分子机制及用药方法选择

目的 研究常用骨肉瘤化疗药物作用的分子机制,对临床用药方式提供指导.方法 将骨肉瘤细胞混悬液与不同浓度吡柔比星、顺铂、卡铂、甲氨蝶呤、异环磷酰胺分别作用24、48、72 h后,用倒置显微镜观察细胞生长情况.用MTT法测定每一孔的吸光度值OD,制作生长曲线,并求出药物的IC50.用IC50浓度的药物分别作用于骨肉瘤细胞24、48和72 h后用流式细胞仪进行细胞周期分析,了解细胞的周期分布及细胞的增殖活性.免疫印迹技术观察细胞的PCNA、Bcl-2、Bax、cyclin D1、cyclin E的表达.结果 在倒置显微镜下观察不同药物浓度作用于的骨肉瘤细胞,比较细胞死亡数量于时间关系.MTT实验:不同时间组及不同浓度组与对照组相比,对MG-63细胞的生长抑制作用差异均具有显著性(P＜0.05),呈现不同剂量-时间效应关系.细胞周期分析:在细胞各期可见明显凋亡峰,细胞周期各期所占比例无明显变化,抑制作用呈剂量依赖性.蛋白免疫印迹:PCNA在不同药物组骨肉瘤表达与对照组相比明显不同,不同药物组Bax表达与对照组明显增加,Bcl-2表达减少.cyclin D1、cyclin E的表达明显减少.结论 吡柔比星对MG-63细胞的抑制率和时间关系密切,和浓度有一定的关系.同样剂量吡柔比星5d的效果比3d好.卡铂具有与顺铂类似的效应,但卡铂如果替代顺铂在化疗中的作用,卡铂的量必须是顺铂的25倍以上.甲氨蝶呤作用于骨肉瘤细胞MG-63后,随着药物浓度的增加和时间的延长,作用逐渐增强,尤其是用量和效果有明显关系.异环磷酰胺在体外对骨肉瘤细胞无明显抑制作用.     

Monoclonal gammopathy (IgA:λ) after autologous T-cell-depleted bone marrow transplantation in a patient with non-Hodgkin's lymphoma

Summary We report a case study of a patient suffering from T-lymphoblastic lymphoma, for whom autologous T-cell-depleted bone marrow transplantation was carried out. Low-dose cyclosporin A (CsA) was administered just after autologous bone marrow transplantation (ABMT) for induction of autologous graft-versus-host disease. Three months later, the lymphoma relapsed with marked monoclonal gammopathy of the immunoglobulin A type, which had not been seen before ABMT. It is suggested that the regrowth of the lymphoma cells was related to the maturation of B cells or the secretion of immunoglobulins from plasma cells, associated with some cytokines produced under the condition of an abnormal immunological circumstance after ABMT plus CsA.     

Basophil activation test: Mechanisms and considerations for use in clinical trials and clinical practice

The basophil activation test (BAT) is a functional assay that measures the degree of degranulation following stimulation with allergen or controls by flow cytometry. It correlates directly with histamine release. From the dose-response curve resulting from BAT in allergic patients, basophil reactivity (%CD63⁺ basophils) and basophil sensitivity (EC₅₀ or similar) are the main outcomes of the test. BAT takes into account all characteristics of IgE and allergen and thus can be more specific than sensitization tests in the diagnosis of allergic disease. BAT reduces the need for in vivo procedures, such as intradermal tests and allergen challenges, which can cause allergic reactions of unpredictable severity. As it closely reflects the patients' phenotype in most cases, it may be used to support the diagnosis of food, venom and drug allergies and chronic urticaria, to monitor the natural resolution of food allergies and to predict and monitor clinical the response to immunomodulatory treatments, such as allergen-specific immunotherapy and biologicals. Clinical application of BAT requires analytical validation, clinical validation, standardization of procedures and quality assurance to ensure reproducibility and reliability of results. Currently, efforts are ongoing to establish a platform that could be used by laboratories in Europe and in the USA for quality assurance and certification.     

MG132对肿瘤恶病质小鼠骨骼肌消耗及TRAF6表达的影响

 目的 探讨蛋白酶体抑制剂MGl32对肿瘤恶病质骨骼肌消耗和TRAF6及其所调节的自噬溶酶体途径和泛素蛋白酶体途径相关因子Beclin-1、MuRF1和MAFbx基因表达的影响。方法 小鼠结肠腺癌C26细胞接种BALB/c小鼠诱导肿瘤恶病质模型。分为对照组（HC）、恶病质组（CC）和MG132治疗组（MG）。监测体质量和自发性活动,于接种后第12天,每天腹腔注射给予MG组小鼠0.1mg/kg的MG132,HC和CC组小鼠给予等量生理盐水,第19天处死小鼠。称量肿瘤、左侧腓肠肌重量,检测腓肠肌横切面积,RT-PCR和Western blot 检测TRAF6、Beclin1、MuRF1和MAFbx的mRNA和蛋白水平。结果 CC组小鼠去瘤体质量、自发性活动、腓肠肌重量和横切面积均显著低于HC组(P<0.05),MG组小鼠这次指标均显著回升(P<0.05),但仍低于HC组(P<0.05)。CC组小鼠腓肠肌组织TRAF6、Beclin1、MAFbx和MuRF1的mRNA和蛋白水平均显著高于HC组(P<0.05),MG组小鼠这次指标均显著低于CC组(P<0.05)。结论 MG132改善肿瘤恶病质骨骼肌消耗可能与抑制腓肠肌TRAF6的表达,阻断自噬溶酶体途径和泛素蛋白酶体途径有关。     

Biofilm and cell adhesion strength on dental implant surfaces via the laser spallation technique

ObjectiveThe aims of this study are to quantify the adhesion strength differential between an oral bacterial biofilm and an osteoblast-like cell monolayer to a dental implant-simulant surface and develop a metric that quantifies the biocompatible effect of implant surfaces on bacterial and cell adhesion.MethodsHigh-amplitude short-duration stress waves generated by laser pulse absorption are used to spall bacteria and cells from titanium substrates. By carefully controlling laser fluence and calibration of laser fluence with applied stress, the adhesion difference between Streptococcus mutans biofilms and MG 63 osteoblast-like cell monolayers on smooth and rough titanium substrates is obtained. The ratio of cell adhesion strength to biofilm adhesion strength (i.e., Adhesion Index) is determined as a nondimensionalized parameter for biocompatibility assessment.ResultsAdhesion strength of 143 MPa, with a 95% C.I. (114, 176), is measured for MG 63 cells on smooth titanium and 292 MPa, with a 95% C.I. (267, 306), on roughened titanium. Adhesion strength for S. mutans on smooth titanium is 320 MPa, with a 95% C.I. (304, 333), and remained relatively constant at 332 MPa, with a 95% C.I. (324, 343), on roughened titanium. The calculated Adhesion Index for smooth titanium is 0.451, with a 95% C.I. (0.267, 0.622), which increased to 0.876, with a 95% C.I. (0.780, 0.932), on roughened titanium.SignificanceThe laser spallation technique provides a platform to examine the tradeoffs of adhesion modulators on both biofilm and cell adhesion. This tradeoff is characterized by the Adhesion Index, which is proposed to aid biocompatibility screening and could help improve implantation outcomes. The Adhesion Index is implemented to determine surface factors that promote favorable adhesion of cells greater than biofilms. Here, an Adhesion Index ? 1 suggests favorable biocompatibility.     

EEC‐ and ADULT‐Associated TP63 Mutations Exhibit Functional Heterogeneity Toward P63 Responsive Sequences

TP63 germ‐line mutations are responsible for a group of human ectodermal dysplasia syndromes, underlining the key role of P63 in the development of ectoderm‐derived tissues. Here, we report the identification of two TP63 alleles, G134V (p.Gly173Val) and insR155 (p.Thr193_Tyr194insArg), associated to ADULT and EEC syndromes, respectively. These alleles, along with previously identified G134D (p.Gly173Asp) and R204W (p.Arg243Trp), were functionally characterized in yeast, studied in a mammalian cell line and modeled based on the crystal structure of the P63 DNA‐binding domain. Although the p.Arg243Trp mutant showed both complete loss of transactivation function and ability to interfere over wild‐type P63, the impact of p.Gly173Asp, p.Gly173Val, and p.Thr193_Tyr194insArg varied depending on the response element (RE) tested. Interestingly, p.Gly173Asp and p.Gly173Val mutants were characterized by a severe defect in transactivation along with interfering ability on two DN‐P63α‐specific REs derived from genes closely related to the clinical manifestations of the TP63‐associated syndromes, namely PERP and COL18A1. The modeling of the mutations supported the distinct functional effect of each mutant. The present results highlight the importance of integrating different functional endpoints that take in account the features of P63 proteins' target sequences to examine the impact of TP63 mutations and the associated clinical variability.     

宫颈鳞状细胞癌组织中p63和p40的表达及其临床意义

目的探讨宫颈鳞状细胞癌组织中p63和p40的表达及其临床意义。方法选择宫颈鳞状细胞癌组织标本83例,采用免疫组织化学染色法检测p63和p40的表达,分析p63和p40在宫颈鳞状细胞癌中的表达状况,并分析宫颈鳞状细胞癌组织中p63和p40表达与患者临床病理特征的关系以及二者之间的相关性。结果在83例宫颈鳞状细胞癌组织中,呈p63弱阳性或阳性表达者65例,阳性率为78.31%,呈p40弱阳性或阳性表达者72例,阳性率为86.75%,二者均呈弱阳性或阳性表达者53例,阳性率为63.86%,二者均呈阴性表达者10例,阴性率为12.05%。不同年龄、部位的宫颈鳞状细胞癌患者组织中p63和p40的阳性表达无明显差异性(χ^2分别=1.17、1.75、0.52、0.71,P均>0.05),但不同肿瘤大小、临床分期、分化程度、肌层浸润深度及有无淋巴结转移、脉管浸润、宫旁浸润的患者组织中p63和p40的阳性表达存在显著性差异(χ^2分别=5.72、23.14、13.10、43.40、53.99、61.36、62.43、6.57、33.73、7.36、35.85、35.87、22.13、50.02,P均<0.05)。Pearson相关性分析显示,宫颈鳞状细胞癌组织中p63与p40的表达水平呈正相关(r=0.96,P<0.05)。结论p63和p40在宫颈鳞状细胞癌组织中具有较高的阳性表达率,与患者的疾病恶性程度有关,且二者表达呈正相关。