转录因子Nkx2.5、ISL-1表达与小鼠胚胎心的早期发育

目的:探讨转录因子Nkx2.5和ISL-1的表达与小鼠胚胎早期心发育的关系.方法:胚龄7.5～13d小鼠胚胎连续石蜡切片,用抗α-平滑肌肌动蛋白(α-SMA)、抗心肌肌球蛋白重链(MHC)、抗转录因子ISL-1、抗Nkx2.5抗体,进行免疫组织化学显色.结果:转录因子Nkx2.5在胚龄7.5d生心板表达早于心肌特异性蛋白MHC及SMA.随心外形建立,Nkx2.5在心各部表达逐渐增强.胚龄9d, Nkx2.5阳性表达从原始心管动脉端延伸至心包腔背侧脏壁中胚层及咽周围轴旁中胚层.Isl-1开始表达于胚龄8d原始心管心背系膜及前肠腹侧脏壁中胚层间充质,阳性细胞数量在11～12d达顶峰,形成前肠腹侧连接流出道远端和心房背侧壁的ISL-1阳性细胞带.11～12d,流出道远端出现Nkx2.5和MHC阴性,ISL-1、SMA阳性表达区.胚龄13d后,Nkx2.5和ISL-1在心和前肠腹侧间充质的表达强度下降.结论:Nkx2.5是原始生心区心肌前体细胞早期分化的标记蛋白,在第2生心区表达限于胚龄9d、咽周脏壁和轴旁中胚层心肌前体细胞亚群.第2生心区标记蛋白ISL-1主要表达在胚龄8～12d前肠腹侧脏壁中胚层心肌前体细胞,并向心管添加心肌细胞,参与流出道发育及心管成襻.11d后,第2生心区间充质细胞开始分化为升主动脉和肺动脉干平滑肌.     

Toll-like receptor agonists shape the immune responses to a mannose receptor-targeted cancer vaccine

Previous studies have documented that selective delivery of protein antigens to cells expressing mannose receptor (MR) can lead to enhanced immune responses. We postulated that agents that influenced the MR expression level, and the activation and migration status of MR-expressing antigen presenting cells, would modulate immune responses to MR-targeted vaccines. To address this question, we investigated the effect of clinically used adjuvants in human MR transgenic (hMR-Tg) mice immunized with an MR-targeting cancer vaccine composed of the human anti-MR monoclonal antibody B11 fused with the oncofetal protein, human chorionic gonadotropin beta chain (hCGβ), and referred to as B11-hCGβ. We found that humoral responses to low doses of B11-hCGβ could be enhanced by prior administration of GM-CSF, which upregulated MR expression in vivo. However, co-administration of the Toll-like receptor (TLR) agonists, poly-ICLC and/or CpG with B11-hCGβ was required to elicit Th1 immunity, as measured by antigen-specific T-cell production of IFN-γ. The TLR agonists were shown to increase the number of vaccine-containing cells in the draining lymph nodes of immunized hMR-Tg mice. In particular, with B11-hCGβ and poly-ICLC, a dramatic increase in vaccine-positive cells was observed in the T-cell areas of the lymph nodes, compared to the vaccine alone or combined with GM-CSF. Importantly, the absence of the TLR agonists during the priming immunization led to antigen-specific tolerance. Therefore, this study provides insight into the mechanisms by which adjuvants can augment immune responses to B11-hCGβ and have implications for the rationale design of clinical studies combining MR-targeted vaccination with TLR agonists.     

WTARLING心脏定律质疑(英文)

本文分析了游离心肌和心肺制备试验结果，并从Starling心脏定律的主要作用、显微结构基础和心脏做功等几个方面进行了讨论。作者认为：①游离心肌条所具有的Frank-Starling效应不能用以表征完整心脏的生理特性，如同一台机器的零件离开整机时所具有的性质不能视为是那台机器的主要性质一样;②在心肺制备试验中增加心脏负荷所引起的心脏收缩活动增强，是心脏具有丰富储备力的表现，它需要通过心肌细胞内的生化机制方能得以实现，因此，根据试验中增加心室充盈压引起心脏收缩活动增强的事实，在逻辑上不应得出心室充盈压力（心肌纤维长度）决定心脏收缩强度的推论;③公式“每搏功＝P×（EDV-ESV）”的数学关系表明，心脏每搏做功和心室舒张末期容积（EDV，心肌纤维初长度）不成正比例或正相关;④Starling心脏定律是一个错误的概念，它不是心脏的一种生理特性。     

Prevention of crescentic glomerulonephritis by immunoneutralization of the fractalkine receptor CX3CR1 rapid communication.

BACKGROUND: Fractalkine is a newly identified T-cell and monocyte/macrophage (Mphi) chemokine with a transmembrane domain and is a cell-surface protein on activated endothelium. It can mediate adhesion of cells expressing the fractalkine receptor CX3CR1. These unique features make fractalkine well suited for leukocyte recruitment in tissues with high blood flow as in the renal glomerulus. METHODS: Fractalkine expression in glomeruli and response of isolated glomerular inflammatory cells to fractalkine were studied in the Wistar-Kyoto (WKY) crescentic glomerulonephritis model. Antibody was used to confirm the proinflammatory role of fractalkine. RESULTS: Fractalkine was markedly induced in the endothelium of nephritic rat glomeruli, and inflammatory leukocytes infiltrating the glomeruli expressed increased levels of CX3CR1. Anti-CX3CR1 antibody treatment dramatically blocked leukocyte infiltration in the glomeruli, prevented crescent formation, and improved renal function. CONCLUSIONS: Fractalkine plays a central role in leukocyte trafficking at the endothelium in the high-flow glomerular circuit and, in turn, implicates CX3CR1 as a prime drug target for therapeutic intervention of endothelium-related inflammatory diseases.     

Applying chemical bile duct embolization to achieve chemical hepatectomy in hepatolithiasis: a further experimental study

Background

Hepatolithiasis is the presence of calculi within the bile ducts of the liver. It represents a significant problem for hepatobiliary surgery because of its high recurrence rate and the associated risk for partial hepatectomy. This study was designed to explore the long-term efficacy of chemical biliary duct embolization (CBDE) to treat recurrent hepatolithiasis.

Materials and methods

A rabbit model of hepatolithiasis was established, and CBDE was achieved using oxybenzene and N-butyl-cyanoacrylate. The short-term (6 wk) and long-term (12 wk) efficacy of CBDE treatment was compared by observing the degree of atrophy, fibrosis, proliferation of collagen fibers, and apoptosis of hepatocytes and hepatic stellate cells in the embolized hepatic lobe. Biochemical measurement of β-glucuronidase was also evaluated to determine the effect of CBDE on stone formation.

Results

Six weeks after CBDE, there was liver cell destruction, collagen accumulation, and bile duct proliferation only in the peripheral part of the target lobe. Twelve weeks after CBDE, “self-cut” chemical hepatectomy was achieved, as manifested by the destruction of almost all the hepatocytes in the target lobe, bile duct proliferation, and collagen fiber accumulation. The β-glucuronidase activity was markedly lower in the embolized lobe than in the nonembolized lobe. In contrast, bax, caspase-3, caspase-9, and α-smooth muscle actin expression was substantially higher in the embolized lobe than in the sham-operation group at 6 wk, but was lower at 12 wk.

Conclusions

CBDE is a potentially effective therapeutic approach for treating and preventing the recurrence of hepatolithiasis.     

脑内移植时星状胶质细胞的变化

本文用胶质纤维酸性蛋白免疫组织化学方法研究了帕金森氏病模型鼠、老年性痴呆模型鼠及皮质损伤鼠等在脑移植２～３个月后移植区及其与宿主交界面的胶质细胞变化。结果发现不同动物模型的胚脑移植区及其与宿主交界面的胶质变化基本相同．Ｎｉｓｓｌ染色切片上可见移植区和宿主之间有直径小于８μｍ的小细胞集聚。胶质纤维酸性蛋白免疫组织化学法发现移植区和宿主脑交界面有深染的阳性纤维及细胞或交界面形成一包膜状结构；还有的在交界面无明显的胶质纤维酸性蛋白阳性纤维分隔，呈现两者相互融合的趋势．移植区内可见两种胶质细胞，一种与正常胶质细胞近似，另一种为胞体大、突起粗且深染的反应性胶质细胞；移植区内的血管周围也见有反应性胶质细胞包绕。本研究表明移植也可引起胶质反应，并且移植区内反应性胶质细胞可能对植入的神经元提供其生存所需要的营养物质。     

Role of mitochondrial inner membrane permeabilization in necrotic cell death,apoptosis, and autophagy

Inhibition of mitochondrial oxidative phosphorylation progresses to uncoupling when opening of cyclosporin A-sensitive permeability transition pores increases permeability of the mitochondrial inner membrane to small solutes. Involvement of the mitochondrial permeability transition (MPT) in necrotic and apoptotic cell death is implicated by demonstrations of protection by cyclosporin A against oxidative stress, ischemia/reperfusion, tumor necrosis factor-alpha exposure, Fas ligation, calcium overload, and a variety of toxic chemicals. Confocal microscopy directly visualizes the MPT in single mitochondria within living cells from the translocation of impermeant fluorophores, such as calcein, across the inner membrane. Simultaneously, mitochondria release potential-indicating fluorophores. Subsequently, mitochondria swell, causing outer membrane rupture and release of cytochrome c and other proapoptotic proteins from the intermembrane space. In situ a sequence of decreased NAD(P)H, increased free calcium, and increased reactive oxygen species formation within mitochondria promotes the MPT and subsequent cell death. Necrotic and apoptotic cell death after the MPT depends, in part, on ATP levels. If ATP levels fall profoundly, glycine-sensitive plasma membrane permeabilization and rupture ensue. If ATP levels are partially maintained, apoptosis follows the MPT. The MPT also signals mitochondrial autophagy, a process that may be important in removing damaged mitochondria. Cellular features of necrosis, apoptosis, and autophagy frequently occur together after death signals and toxic stresses. A new term, necrapoptosis, describes such death processes that begin with a common stress or death signal, progress by shared pathways, but culminate in either cell lysis (necrosis) or programmed cellular resorption (apoptosis), depending on modifying factors such as ATP.     

单倍体相合异基因造血干细胞移植治疗白血病

背景：对于无HLA全相合同胞供者的患者,采用单倍体相合造血干细胞移植面临移植物抗宿主病重、移植相关死亡率高的风险,但通过不同的移植模式,将有可能获取相近的疗效。 目的：观察亲缘HLA单倍体相合异基因造血干细胞移植治疗白血病的疗效,并与亲缘HLA全相合异基因造血干细胞移植相比较。 方法：45例白血病患者分为2组。单倍体组移植方式为外周血或联合骨髓干细胞移植,预处理方案为改良白消安与环磷酰胺或加抗胸腺细胞球蛋白,移植物抗宿主病的预防采用环孢素A+甲氨蝶呤+霉酚酸脂;全相合组移植方式为外周血干细胞移植,预处理方案为BuCY,移植物抗宿主病的预防采用环孢素A+甲氨蝶呤。 结果与结论：两组均获得造血重建时间差异无显著性意义。单倍体及全相合组急性移植物抗宿主病的累积发病率分别为73%对52%(P > 0.05);慢性移植物抗宿主病的累积发病率分别为56%对45%(P > 0.05);移植相关死亡率分别为36%对17%(P > 0.05);单倍体组无复发,全相合组复发2例;两组的预计3年累积无病生存率分别为61%对60%(P > 0.05)。结果提示,亲缘单倍体异基因造血干细胞移植的总体疗效与亲缘全相合异基因造血干细胞移植相似,但中重度急性移植物抗宿主病的发生率较后者为高。     

电化学治疗昆明小鼠肉瘤及其机理分析

研究了电化学治疗昆明小鼠肉瘤的疗效,并分析其机理。在体外将S-180细胞用不同参数的电场处理,研究适合电化学治疗的电场条件。通过复制肉瘤模型,将肉瘤小鼠随机分成4组:对照组、电疗组、化疗组、电化疗组。研究了不同处理组的肿瘤抑瘤率、治愈率以及小鼠的自由基代谢水平。结果电化疗组的抑瘤率、治愈率都显著高于化疗组和电疗组(P<0.05),电化疗组小鼠受到氧自由基的攻击显著降低,免疫力提高。分析机理发现,电化学治疗肿瘤的机理可能至少涉及细胞膜通透性提高、细胞的耐药性降低、机体的免疫力提高三个方面。     

军人心理健康教育骨干培训模式探讨

通过对军人心理健康教育骨干培训的理论和实践探索，提出了军人心理健康教育骨干培训的基本模式——“一个目标、二个关键、三个阶段、四种途径、五项内容”的整合培训模式。①一个目标：培训部队实用型心理健康教育骨干；②两个关键：理论与实际结合，院校与部队结合；③三个阶段：军人心理健康教育知识技能培训阶段、心理健康评估和干预培训阶段、心理健康教育知识技能应用实践阶段；④四种途径：理论学习、实习操作、临床见习、部队实践；⑤五项内容：心理健康的评估和心理档案建立、心理障碍的识别与诊断、军人心理健康讲座、心理健康教育报刊、个别与团体咨询。