• 【右精索静脉曲张和缺氧,是男性不育症的关键因素:睾丸引流系统受损的流体力学分析。】 复制标题 收藏 收藏
    DOI:10.1016/s1472-6483(10)60638-4 复制DOI
    作者列表:Gat Y,Gornish M,Navon U,Chakraborty J,Bachar GN,Ben-Shlomo I
    BACKGROUND & AIMS: :Varicocele is considered a predominantly unilateral left-sided disease. However, since male fertility is preserved with only one healthy testis, infertility perforce represents bilateral testicular dysfunction. It was hypothesized that: (i) right varicocele cannot be diagnosed by palpation and therefore has not been treated in the past by the traditional treatment, and (ii) right varicocele causes impaired oxygen supply in the right testicular microcirculation, leading to germ cell degeneration. This study performed venographies of both right and left internal spermatic veins during the treatment of 840 infertile men with varicocele and analysed the results using tools of fluid mechanics. Histopathology of the right testis revealed stagnation of blood flow and degenerative changes attributed to lack of adequate oxygenation in all testicular cell types. Right varicocele was found in the vast majority of the patients. We found that due to the destruction of one-way valves, pathologic hydrostatic pressure is produced in the testicular venous microcirculatory system about five times higher than normal, exceeding arteriolar pressure. The pressure gradient between the arterioles and venules in the testicular tissue is therefore reversed, leading to persistent hypoxia. Right varicocele, although undetected, is prevalent in infertile men with varicocele, hence only bilateral occlusion of the internal spermatic veins, including the associated bypasses, eliminating the pathologic hydrostatic pressure will lead to resumption of arterial blood flow in the testicular microcirculation.
    背景与目标: :精索静脉曲张被认为是主要的单侧左侧疾病。但是,由于只有一个健康的睾丸才能维持男性的生育能力,因此不育症的强迫表现为双侧睾丸功能障碍。假设:(i)右精索静脉曲张不能通过触诊诊断,因此过去没有用传统疗法治疗;(ii)右精索静脉曲张导致右睾丸微循环中的氧气供应受损,导致生殖细胞变性。这项研究在治疗840名不育男性精索静脉曲张的过程中对左右两侧的精索静脉进行了静脉造影,并使用流体力学工具对结果进行了分析。右睾丸的组织病理学显示血流停滞和变性变化归因于所有睾丸细胞类型缺乏足够的氧合。在绝大多数患者中发现了右精索静脉曲张。我们发现由于单向阀的破坏,睾丸静脉微循环系统中产生的病理性静水压力比正常高约五倍,超过了小动脉压力。因此,睾丸组织中小动脉和小静脉之间的压力梯度会反向,从而导致持续的缺氧。右精索静脉曲张虽然未被发现,但在精索静脉曲张的不育男性中很普遍,因此,只有双侧内精子静脉闭塞,包括相关的旁路,消除病理性静水压会导致睾丸微循环中动脉血流的恢复。
  • 【大鼠角膜缘和中央角膜上皮中基因表达(SAGE)的系列分析。】 复制标题 收藏 收藏
    DOI:10.1167/iovs.06-0216 复制DOI
    作者列表:Adachi W,Ulanovsky H,Li Y,Norman B,Davis J,Piatigorsky J
    BACKGROUND & AIMS: PURPOSE:To identify genes preferentially expressed in the stem-cell-rich limbal epithelium of the rat cornea. METHODS:The limbal and central corneal epithelial cells of 6-week-old rats were isolated by microdissection. Serial analysis of gene expression (SAGE) libraries were constructed and analyzed, and in situ hybridization, reverse transcription-polymerase chain reaction (RT-PCR) and cDNA cloning were conducted by conventional procedures. RESULTS:The rat limbal and central corneal epithelial SAGE libraries consisted of 41,894 and 40,691 tags, respectively. After annotation, this was reduced to 759 transcripts specific for the limbal library and 844 transcripts specific for the central corneal library; 2292 transcripts overlapped. Transcripts encoding proteins with metabolic functions comprised the major functional category in both libraries. In situ hybridization and/or RT-PCR results of 12 of the most abundant, highly enriched transcripts in the limbal epithelium were in general agreement with the SAGE data and showed that these proteins are also expressed in the conjunctival epithelium. Interesting limbal-enriched transcripts encode WDNM1-like protein (similar to WDNM1/Expi, a putative secreted proteinase and inhibitor of metastasis), mesothelin (a cancer marker), marapsin (a trypsin-like serine protease that may control cell growth and migration), K4 and K15 (both cytokeratins), and membrane-spanning four-domain subfamily A member 8B. WDNM1-like protein was cloned and confirmed as a member of the four-disulfide core family. CONCLUSIONS:The SAGE results extend the database of genes expressed in the rodent cornea and suggest an association between several genes preferentially expressed in the limbal epithelium with cellular proliferation and migration.
    背景与目标: 目的:鉴定在大鼠角膜的富含干细胞的角膜缘上皮细胞中优先表达的基因。
    方法:采用显微解剖技术分离6周龄大鼠角膜缘和角膜上皮细胞。构建并分析基因表达(SAGE)库的序列分析,并通过常规方法进行原位杂交,逆转录-聚合酶链反应(RT-PCR)和cDNA克隆。
    结果:大鼠角膜缘和角膜上皮SAGE文库分别由41,894和40,691个标签组成。注释后,该数目减少为角膜缘文库特异的759个转录物和中央角膜文库特异的844个转录物。 2292个成绩单重叠。编码具有代谢功能的蛋白质的转录本在两个文库中均属于主要功能类别。角膜缘上皮细胞中12种最丰富,高度富集的转录本的原位杂交和/或RT-PCR结果与SAGE数据基本一致,表明这些蛋白也在结膜上皮细胞中表达。有趣的角膜缘丰富的转录本编码WDNM1样蛋白(类似于WDNM1 / Expi,一种推测的分泌蛋白酶和转移抑制剂),间皮素(一种癌症标志物),marapsin(一种胰蛋白酶样丝氨酸蛋白酶,可以控制细胞的生长和迁移)。 ,K4和K15(均为细胞角蛋白)和跨膜四结构域亚家族A成员8B。克隆了类似WDNM1的蛋白质,并确认其为四-二硫键核心家族的成员。
    结论:SAGE结果扩展了在啮齿动物角膜中表达的基因数据库,并暗示了在角膜缘上皮中优先表达的几个基因与细胞增殖和迁移之间的关联。
  • 【艾滋病患者的更昔洛韦耐药性巨细胞病毒(CMV)视网膜炎一例:CMV病毒载量和血室中病毒突变的纵向分子分析。】 复制标题 收藏 收藏
    影响因子 :
    发表时间:1997-06-01
    来源期刊:AIDS
    DOI:10.1097/00002030-199707000-00005 复制DOI
    作者列表:Boivin G,Gilbert C,Morissette M,Handfield J,Goyette N,Bergeron MG
    BACKGROUND & AIMS: OBJECTIVE:To study the temporal relationships between cytomegalovirus (CMV) viral load and specific UL97 mutations in polymorphonuclear leukocytes (PMNL) and plasma samples from a patient with AIDS who developed ganciclovir-resistant CMV retinitis.

    METHODS:Sequential PMNL and plasma samples were analysed for determination of the CMV viral load using non-molecular methods and a quantitative polymerase chain reaction (PCR) assay. Screening of the same samples for the most common mutations conferring ganciclovir resistance was performed using nested PCR and restriction enzyme analysis.

    RESULTS:At the time of progression of CMV retinitis (after 6 months of ganciclovir), a rapid increase in the CMV DNA load was found in both PMNL and plasma samples. This increase paralleled the emergence of a specific mutation (V594) in the same samples and recovery of ganciclovir-resistant blood isolates. In this patient, however, the only tests that substantially predicted the progression of CMV disease were the quantitative PCR assay using PMNL and to a lesser extent the pp65 antigenemia assay.

    CONCLUSIONS:Quantitative evaluation of the CMV viral load in PMNL using sensitive assays such as PCR appears to be a promising approach for monitoring antiviral therapy in subjects with AIDS. In addition, common mutations conferring ganciclovir resistance can be detected directly in PMNL and plasma samples.

    背景与目标: 目标:研究巨细胞病毒耐药的巨细胞病毒性视网膜炎的艾滋病患者的巨细胞病毒(CMV)病毒载量与多形核白细胞(PMNL)和血浆样品中特定UL97突变之间的时间关系。

    方法:使用非分子方法和定量聚合酶链反应(PCR)分析了连续的PMNL和血浆样品,以测定CMV病毒载量。使用巢式PCR和限制性内切酶分析对相同样品中最引起更昔洛韦耐药的突变进行筛查。

    结果:在CMV视网膜炎进展时(6个月后) (更昔洛韦)的检测,在PMNL和血浆样品中均发现CMV DNA载量迅速增加。这种增加与在相同样品中出现特定突变(V594)以及对更昔洛韦耐药的血液分离株的回收率平行。然而,在该患者中,唯一可以预测CMV疾病进展的检测方法是使用PMNL进行定量PCR检测,而在较小程度上使用pp65抗原血症检测。

    结论:定量使用诸如PCR的敏感测定法评估PMNL中CMV病毒载量似乎是监测艾滋病患者抗病毒治疗的一种有前途的方法。此外,可以在PMNL和血浆样品中直接检测到更昔洛韦耐药的常见突变。

  • 【结肠扩张激活蓝藻:促肾上腺皮质激素释放因子和兴奋性氨基酸的作用。】 复制标题 收藏 收藏
    DOI:10.1016/s0006-8993(97)00116-9 复制DOI
    作者列表:Lechner SM,Curtis AL,Brons R,Valentino RJ
    BACKGROUND & AIMS: The present study was designed to elucidate the neurotransmitters involved in activation of the noradrenergic nucleus, locus coeruleus, by distention of the distal colon. Locus coeruleus spontaneous discharge rate was recorded from halothane-anesthetized rats before, during and after distention of the colon produced by inflation of a balloon catheter with varying volumes of water. Locus coeruleus activation by colon distention was volume-dependent and reversible. Activation of cortical electroencephalographic activity was temporally correlated with locus coeruleus activation during colon distention and prolonged distention (greater than 2 min) resulted in tachyphalaxis to both locus coeruleus and cortical electroencephalographic activation. The corticotropin-releasing factor antagonist, DPheCRF(12-41), administered intracerebroventricularly (3 microg) or microinfused into the locus coeruleus (10 ng) significantly attenuated locus coeruleus activation produced by lower, but not higher magnitudes of colon distention, implicating corticotropin-releasing factor afferents to the locus coeruleus in this response. Consistent with this, prior exposure to 30 min of footshock stress, which desensitizes locus coeruleus neurons to corticotropin-releasing factor, produced a similar attenuation of locus coeruleus activation by low, but not high magnitudes of distention. Kynurenic acid, administered intracerebroventricularly (5 micromol), significantly antagonized locus coeruleus activation by all magnitudes of colon distention. However, this excitatory amino acid antagonist was ineffective when administered directly into the locus coeruleus (0.3 nmol). Together, these findings suggest that low magnitudes of colon distention activate the locus coeruleus-noradrenergic system via corticotropin-releasing factor release within the locus coeruleus and that excitatory amino acid neurotransmission at a site distal to the locus coeruleus is necessary for this response. Activation of the locus coeruleus-noradrenergic system during colon distention may serve as a cognitive limb of the peripheral parasympathetic response. This activation may also play a role in disorders characterized by comorbidity of colonic and psychiatric symptoms, such as irritable bowel syndrome.

    背景与目标: 本研究旨在阐明远端结肠扩张引起的去甲肾上腺素能核激活的神经递质。记录了氟烷麻醉大鼠在用不同体积的水充胀球囊导管充盈结肠之前,期间和之后自发排出蓝藻的频率。结肠扩张引起的蓝斑轨迹激活是体积依赖性的和可逆的。皮质脑电图活动的激活在时间上与结肠扩张期间脑蓝素的激活相关,而长期扩张(大于2分钟)会导致脑脊髓蓝素和皮质脑电图的速动性。皮质激素释放因子拮抗剂DPheCRF(12-41),经脑室内(3微克)或微输注到蓝斑(10 ng)时,可显着减弱由较低但不是较高程度的结肠扩张产生的蓝斑激活,这暗示促肾上腺皮质激素-在这种反应中,释放因子传入蓝斑。与此相一致,事先暴露于30分钟的足底震荡中,使蓝斑蓝皮病的神经元对促肾上腺皮质激素释放因子不敏感,通过低但不高的扩张程度,蓝斑蓝皮病的激活也有类似的减弱。脑室内(5微摩尔)施用的犬尿酸通过各种程度的结肠扩张显着拮抗蓝斑轨迹激活。但是,这种兴奋性氨基酸拮抗剂当直接施用于蓝斑(0.3 nmol)时无效。总之,这些发现表明低水平的结肠扩张通过蓝藻中的促肾上腺皮质激素释放因子释放来激活蓝藻-去甲肾上腺素能系统,并且在蓝藻远侧的部位,兴奋性氨基酸神经传递对于该反应是必需的。结肠扩张期间蓝斑-去甲肾上腺素能系统的激活可能充当外周副交感反应的认知肢体。这种活化作用还可能在以结肠和精神症状合并症为特征的疾病中起作用,例如肠易激综合症。

  • 【缺氧条件下的肿瘤基质细胞相互作用通过肝细胞生长因子/ c-Met途径增加了胰腺癌细胞的侵袭性。】 复制标题 收藏 收藏
    DOI:10.1002/ijc.22178 复制DOI
    作者列表:Ide T,Kitajima Y,Miyoshi A,Ohtsuka T,Mitsuno M,Ohtaka K,Koga Y,Miyazaki K
    BACKGROUND & AIMS: :The hypoxic environment in tumor is reported to play an important role in pancreatic cancer progression. The interaction between stromal and cancer cells also contributes to the malignant behavior of pancreatic cancer. In the present study, we investigated whether hypoxic stimulation affects stromal as well as pancreatic cancer cells. Our findings demonstrated that hypoxia remarkably elevated the HIF-1alpha expression in both pancreatic cancer (PK8) and fibroblast cells (MRC5). Hypoxic stimulation accelerated the invasive activity of PK8 cells, and invasiveness was thus further accelerated when the hypoxic PK8 cells were cultured with conditioned medium prepared from hypoxic MRC5 cells (hypoxic conditioned medium). MMP-2, MMP-7, MT1-MMP and c-Met expressions were increased in PK8 cells under hypoxia. Hypoxic stimulation also increased the hepatocyte growth factor (HGF) secretion from MRC5 cells, which led to an elevation of c-Met phosphorylation in PK8 cells. Conversely, the elevated cancer invasion, MMP activity and c-Met phosphorylation of PK8 cells were reduced by the removal of HGF from hypoxic conditioned medium. In immunohistochemical study, the HIF-1alpha expression was observed in surrounding stromal as well as pancreatic cancer cells, thus indicating hypoxia exists in both of cancer and stromal cells. Moreover, the stromal HGF expression was found to significantly correlate with not only the stromal HIF-1alpha expression but also the c-Met expression in cancer cells. These results indicate that the hypoxic environment within stromal as well as cancer cells activates the HGF/c-Met system, thereby contributing to the aggressive invasive features of pancreatic cancer.
    背景与目标: :据报道肿瘤中的低氧环境在胰腺癌的进展中起重要作用。基质细胞与癌细胞之间的相互作用也有助于胰腺癌的恶性行为。在本研究中,我们调查了低氧刺激是否影响基质以及胰腺癌细胞。我们的发现表明,低氧显着提高了胰腺癌(PK8)和成纤维细胞(MRC5)中HIF-1alpha的表达。低氧刺激加速了PK8细胞的侵袭活性,因此,当用由低氧MRC5细胞制备的条件培养基(低氧条件培养基)培养低氧PK8细胞时,侵袭性进一步加快。在缺氧条件下,PK8细胞中的MMP-2,MMP-7,MT1-MMP和c-Met表达增加。缺氧刺激还增加了MRC5细胞的肝细胞生长因子(HGF)分泌,从而导致PK8细胞中c-Met磷酸化的升高。相反,通过从低氧条件培养基中去除HGF,可以降低PK8细胞的癌浸润,MMP活性和c-Met磷酸化水平的升高。在免疫组织化学研究中,在周围的基质细胞和胰腺癌细胞中均观察到了HIF-1alpha的表达,因此表明在癌细胞和基质细胞中均存在缺氧。此外,发现基质HGF表达不仅与癌细胞中的基质HIF-1α表达而且与c-Met表达显着相关。这些结果表明基质以及癌细胞内的低氧环境激活了HGF / c-Met系统,从而促进了胰腺癌的侵袭性侵袭性。
  • 【胃癌中pRb2 / p130,VEGF,EZH2,p53,p16(INK4A),p27(KIP1),p21(WAF1),Ki-67表达模式的免疫组织化学分析。】 复制标题 收藏 收藏
    DOI:10.1002/jcp.20833 复制DOI
    作者列表:Mattioli E,Vogiatzi P,Sun A,Abbadessa G,Angeloni G,D'Ugo D,Trani D,Gaughan JP,Vecchio FM,Cevenini G,Persiani R,Giordano A,Claudio PP
    BACKGROUND & AIMS: :Although the considerable progress against gastric cancer, it remains a complex lethal disease defined by peculiar histological and molecular features. The purpose of the present study was to investigate pRb2/p130, VEGF, EZH2, p53, p16(INK4A), p27(KIP1), p21(WAF1), Ki-67 expressions, and analyze their possible correlations with clinicopathological factors. The expression patterns were examined by immunohistochemistry in 47 patients, 27 evaluated of intestinal-type, and 20 of diffuse-type, with a mean follow up of 56 months and by Western blot in AGS, N87, KATO-III, and YCC-2, -3, -16 gastric cell lines. Overall, stomach cancer showed EZH2 correlated with high levels of p53, Ki-67, and cytoplasmic pRb2/p130 (P < 0.05, and P < 0.01, respectively). Increased expression of EZH2 was found in the intestinal-type and correlated with the risk of distant metastasis (P < 0.05 and P < 0.01, respectively), demonstrating that this protein may have a prognostic value in this type of cancer. Interestingly, a strong inverse correlation was observed between p27(KIP1) expression levels and the risk of advanced disease and metastasis (P < 0.05), and a positive correlation between the expression levels of p21(WAF1) and low-grade (G1) gastric tumors (P < 0.05), confirming the traditionally accepted role for these tumor-suppressor genes in gastric cancer. Finally, a direct correlation was found between the expression levels of nuclear pRb2/p130 and low-grade (G1) gastric tumors that was statistically significant (P < 0.05). Altogether, these data may help shed some additional light on the pathogenetic mechanisms related to the two main gastric cancer histotypes and their invasive potentials.
    背景与目标: :尽管在对抗胃癌方面取得了长足的进步,但它仍然是由特殊的组织学和分子特征定义的复杂致死性疾病。本研究的目的是研究pRb2 / p130,VEGF,EZH2,p53,p16(INK4A),p27(KIP1),p21(WAF1),Ki-67的表达,并分析其与临床病理因素的可能关系。通过免疫组织化学方法对47例患者的表达模式进行了检查,其中27例为肠型,20例为弥漫型,平均随访56个月,并通过Western blot检测AGS,N87,KATO-III和YCC-2 ,-3,-16个胃细胞系。总体而言,胃癌显示EZH2与高水平的p53,Ki-67和细胞质pRb2 / p130相关(分别为P <0.05和P <0.01)。在肠型中发现EZH2表达增加,并且与远处转移的风险相关(分别为P <0.05和P <0.01),表明该蛋白可能在这种类型的癌症中具有预后价值。有趣的是,观察到p27(KIP1)表达水平与晚期疾病和转移的风险之间存在强烈的负相关(P <0.05),而p21(WAF1)和低度胃癌(G1)的表达水平之间呈正相关。肿瘤(P <0.05),证实了这些肿瘤抑制基因在胃癌中的传统接受作用。最后,在核pRb2 / p130的表达水平与低度(G1)胃肿瘤的表达水平之间发现了直接相关性,具有统计学意义(P <0.05)。总之,这些数据可能有助于进一步阐明与两种主要胃癌组织学类型及其侵袭潜能有关的致病机制。
  • 【葡萄膜炎患者肝素表面改性人工晶状体与常规聚甲基丙烯酸甲酯人工晶状体的回顾性分析。】 复制标题 收藏 收藏
    DOI:10.1007/BF02583275 复制DOI
    作者列表:Lardenoye CW,van der Lelij A,Berendschot TT,Rothova A
    BACKGROUND & AIMS: BACKGROUND:Several studies described the benefits of the heparin-surface-modified intraocular tens (HSM IOL) with regard to the reduced inflammation in routine extracapsular cataract extractions. However, limited information is available about the advantages of the HSM IOL in patients with an intraocular inflammation. AIM:To assess the eventual benefits of the HSM IOL compared to the regular polymethylmethacrylate intraocular lens (PMMA IOL) in patients with uveitis. METHODS:A retrospective study of 43 patients with uveitis of various origins who underwent an extracapsular cataract extraction (24 with HSM, 19 with PMMA IOL). The activity of intraocular inflammation, visual acuity, eventual complications, and medications were examined. Standardized follow-up dates were used (before surgery, one and fourteen days, five and eleven months after surgery.) RESULTS:No difference in the inflammatory, activity was noted between HSM and PMMA groups; neither at short term clinical evaluation, nor at five months after surgery. Despite a slightly better visual acuity in the HSM group before surgery, no long term differences were observed. After surgery the increase in visual acuity was similar for both groups, as well as the frequency of cystoid macular oedema (CMO) and synechiae. Fewer patients in HSM group required Nd:YAG posterior capsulotomy, but the difference was not significant. CONCLUSION:No clinical advantage was found when the HSM IOL was compared with the regular PMMA IOL in 43 patients with uveitis.
    背景与目标: 背景:多项研究描述了肝素表面修饰眼内张力(HSM IOL)在常规囊外白内障摘除术中减少炎症方面的益处。但是,关于眼内炎症患者中HSM IOL的优势的信息很少。
    目的:评估与常规聚甲基丙烯酸甲酯人工晶状体(PMMA IOL)相比,HSM IOL在葡萄膜炎患者中的最终益处。
    方法:回顾性研究43例不同来源葡萄膜炎患者的囊外白内障摘除术(HSM 24例,PMMA IOL 19例)。检查眼内炎症,视力,最终并发症和药物的活动。使用标准化的随访日期(手术前,手术后的第1天,第14天,手术后的第5个月和第11个月)。
    结果:HSM和PMMA组之间在炎症,活性方面没有差异;既不进行短期临床评估,也不在术后五个月进行评估。尽管术前HSM组的视敏度稍好,但未观察到长期差异。手术后,两组的视敏度增加,囊状黄斑水肿(CMO)和粘连的发生率相似。 HSM组需要Nd:YAG后囊切开术的患者较少,但差异无统计学意义。
    结论:将HSM IOL与常规PMMA IOL相比较在43例葡萄膜炎患者中未发现临床优势。
  • 【骨髓嵌合体小鼠的肿瘤浸润基质细胞的制备和功能分析。】 复制标题 收藏 收藏
    DOI:10.1111/j.1348-0421.2006.tb03830.x 复制DOI
    作者列表:Ishigaki H,Yamamoto Y,Ishida H,Kajino K,Itoh Y,Fujiyama Y,Ogasawara K
    BACKGROUND & AIMS: :Tumor-infiltrating stroma cells (TISC) as well as tumors themselves are thought to be involved in tumor-related immunosuppression, which is one of the critical mechanisms of tumor escape from immune surveillance. However, preparation of TISC is difficult because of the small proportion of TISC in established tumors. Thus, the cells thought to be involved in tumor-related immunosuppression are generally prepared from spleens or draining lymph nodes in tumor-bearing mice. In this study, we developed a method for directly preparing TISC from established tumors in order to analyze their function. Using green fluorescent protein (GFP) transgenic (Tg) mice and C57BL/6 mice transplanted with bone marrow (BM) cells of GFPTg mice, we detected three subpopulations of TISC: one is compatible with immature myeloid cells (ImC) derived from BM and the two other subpopulations, CD11b(+) cells and CD11b(-) cells, do not originate from BM. The TISC including these subpopulations but not each subpopulation independently after culturing with tumors in the presence of GM-CSF could suppress T cell proliferation induced by anti-CD3. In our system, tumors did not inhibit T cell responses directly, but unknown factors from tumors affected immunosuppression by TISC.
    背景与目标: :肿瘤浸润基质细胞(TISC)以及肿瘤本身都被认为与肿瘤相关的免疫抑制有关,这是肿瘤逃避免疫监视的关键机制之一。然而,由于在已建立的肿瘤中TISC的比例很小,因此TISC的制备是困难的。因此,通常被认为与肿瘤相关的免疫抑制有关的细胞是从荷瘤小鼠的脾脏或引流淋巴结中制备的。在这项研究中,我们开发了一种从已建立的肿瘤中直接制备TISC的方法,以分析其功能。使用绿色荧光蛋白(GFP)转基因(Tg)小鼠和移植有GFPTg小鼠骨髓(BM)细胞的C57BL / 6小鼠,我们检测到TISC的三个亚群:一个与源自BM的未成熟髓样细胞(ImC)相容。其他两个亚群CD11b()细胞和CD11b(-)细胞并非源自BM。在存在GM-CSF的情况下与肿瘤培养后,TISC包括这些亚群,但并非每个亚群独立地抑制由抗CD3诱导的T细胞增殖。在我们的系统中,肿瘤并未直接抑制T细胞反应,但来自肿瘤的未知因素影响了TISC的免疫抑制。
  • 【产气荚膜梭菌的α-毒素不是鸡坏死性肠炎的必需毒力因子。】 复制标题 收藏 收藏
    DOI:10.1128/IAI.00806-06 复制DOI
    作者列表:Keyburn AL,Sheedy SA,Ford ME,Williamson MM,Awad MM,Rood JI,Moore RJ
    BACKGROUND & AIMS: :The Clostridium perfringens alpha-toxin has previously been implicated as the major virulence factor in necrotic enteritis in chickens, although definitive proof has not been reported. In this study an alpha-toxin mutant was constructed in a virulent chicken isolate and shown to retain full virulence in a chicken disease model. These results demonstrated that alpha-toxin is not an essential virulence factor in the pathogenesis of necrotic enteritis in chickens.
    背景与目标: 产气荚膜梭菌α毒素以前被认为是鸡坏死性肠炎的主要毒力因子,尽管尚未有确切的证据报道。在这项研究中,在有毒的鸡隔离株中构建了α-毒素突变体,并显示在鸡疾病模型中保留了全部毒力。这些结果表明,α-毒素不是鸡坏死性肠炎发病机理中必不可少的毒力因子。
  • 【改良的摆锤装置的生物力学-力系统的理论考虑和体外分析。】 复制标题 收藏 收藏
    DOI:10.1093/ejo/cjl028 复制DOI
    作者列表:Kinzinger GS,Diedrich PR
    BACKGROUND & AIMS: :The aim of this study was to analyse the acting forces and moments induced by a special orthodontic appliance, the Pendulum K, for molar distalization in the transverse and sagittal planes. The purpose-designed test set-up (artificial maxilla with anchorage unit and two electrothermodynamic molars, an electronic measuring unit, a unit with force-moment sensor, an analogue/digital converter, and a data read-out unit) allowed simulation of in vivo conditions on the one hand and precise determination of the force systems on the other. The appliances investigated were three specimens of the Pendulum K. In vitro measurement of the resulting force systems revealed that the forces and moments in the transverse and sagittal planes remained almost constant over a 3 mm measuring increment when the distal screw was continuously activated (10 activations/mm). Without reactivation of the incorporated distal screw, however, a marked decline in the force systems was recorded. The Pendulum K allows translatory distalization of the upper molars and thus dental arch expansion, dispensing with the need for permanent teeth to be extracted, subject to a corresponding indication. This is achieved by continuous adjustment of an incorporated distal screw and by specific pre-activations of the Pendulum springs.
    背景与目标: :这项研究的目的是分析由特殊的正畸矫治器Pendulum K引起的作用力和力矩,以在横断面和矢状面上进行磨牙远侧。专门设计的测试装置(带有锚固单元和两个电热磨牙的人工上颌,一个电子测量单元,一个带有力矩传感器的单元,一个模拟/数字转换器和一个数据读出单元)允许模拟一方面是体内条件,另一方面是精确确定力系统。所研究的器具是Pendulum K的三个标本。对最终产生的力系统进行的体外测量显示,当连续激活远端螺钉(10次激活)时,横向和矢状面的力和力矩在3 mm的测量增量内几乎保持恒定。 /毫米)。然而,在没有重新激活所结合的远端螺钉的情况下,记录了力系统的显着下降。摆K允许上颌臼齿的平移远侧移动,从而使牙弓扩张,并在需要相应指示的情况下无需拔出恒牙。这是通过不断调整内置的远端螺钉并通过对摆弹簧进行特定的预激活来实现的。
  • 【多发性骨髓瘤中热休克蛋白90(HSP90)的表达和HSP90抑制剂(17-AAG)的作用分析。】 复制标题 收藏 收藏
    DOI:10.1080/10428190500472123 复制DOI
    作者列表:Duus J,Bahar HI,Venkataraman G,Ozpuyan F,Izban KF,Al-Masri H,Maududi T,Toor A,Alkan S
    BACKGROUND & AIMS: :Heat shock protein 90 (HSP90) is required for structural folding and maintenance of conformational integrity of various proteins, including several associated with cellular signaling. Recent studies utilizing 17-allylamino-17-demethoxygeldanamycin (17-AAG), an inhibitor of HSP90, demonstrated an antitumor effect in solid tumors. To test whether HSP90 could be targeted in multiple myeloma (MM) patients, we first investigated expression of HSP90 by immunofluorescence and flow cytometric analysis in a myeloma cell line (U266) and primary myeloma cells. Following demonstration of HSP90 expression in myeloma cells, archival samples of 32 MM patients were analysed by immunoperoxidase staining. Myeloma cells in all patients showed strong cytoplasmic expression of HSP90 in all samples and 55% also demonstrated concurrent nuclear immunopositivity. Treatment of U266 and primary MM cells with 17AAG resulted in significantly increased apoptosis compared to untreated control cells. Analysis of anti-apoptotic BCL2 family proteins and akt in MM cells incubated with 17-AAG revealed down-regulation of BCL-2, BCL-XL, MCL-1 and akt. Furthermore, although a low concentration of bortezomib resulted in no cell death, a combination of 17AAG and bortezomib treatment revealed a synergistic apoptotic effect on the U266 cell line. These data suggest that targeted inhibition of HSP90 may prove to be a valid and innovative strategy for the development of future therapeutic options for MM patients.
    背景与目标: :热休克蛋白90(HSP90)是结构折叠和维持各种蛋白质(包括与细胞信号相关的几种蛋白质)构象完整性的必需。利用HSP90抑制剂17-烯丙基氨基-17-去甲氧基格尔德霉素(17-AAG)的最新研究表明,在实体瘤中具有抗肿瘤作用。为了测试HSP90是否可以靶向于多发性骨髓瘤(MM)患者,我们首先通过免疫荧光和流式细胞术分析了骨髓瘤细胞系(U266)和原发性骨髓瘤细胞中HSP90的表达。在证明HSP90在骨髓瘤细胞中表达后,通过免疫过氧化物酶染色分析了32例MM患者的档案样本。在所有患者中,骨髓瘤细胞在所有样品中均表现出强烈的HSP90细胞质表达,并且55%的患者还表现出并发的核免疫阳性。与未处理的对照细胞相比,用17AAG处理U266细胞和原代MM细胞可导致凋亡明显增加。分析与17-AAG孵育的MM细胞中的抗凋亡BCL2家族蛋白和akt,表明BCL-2,BCL-XL,MCL-1和akt下调。此外,尽管低浓度的硼替佐米不会导致细胞死亡,但是17AAG和硼替佐米治疗的组合显示出对U266细胞系的协同凋亡作用。这些数据表明,针对HSP90的靶向抑制可能被证明是开发MM患者未来治疗选择的有效且创新的策略。
  • 【卫生干预措施的优先重点设定:需要进行多标准决策分析。】 复制标题 收藏 收藏
    DOI:10.1186/1478-7547-4-14 复制DOI
    作者列表:Baltussen R,Niessen L
    BACKGROUND & AIMS: :Priority setting of health interventions is often ad-hoc and resources are not used to an optimal extent. Underlying problem is that multiple criteria play a role and decisions are complex. Interventions may be chosen to maximize general population health, to reduce health inequalities of disadvantaged or vulnerable groups, ad/or to respond to life-threatening situations, all with respect to practical and budgetary constraints. This is the type of problem that policy makers are typically bad at solving rationally, unaided. They tend to use heuristic or intuitive approaches to simplify complexity, and in the process, important information is ignored. Next, policy makers may select interventions for only political motives. This indicates the need for rational and transparent approaches to priority setting. Over the past decades, a number of approaches have been developed, including evidence-based medicine, burden of disease analyses, cost-effectiveness analyses, and equity analyses. However, these approaches concentrate on single criteria only, whereas in reality, policy makers need to make choices taking into account multiple criteria simultaneously. Moreover, they do not cover all criteria that are relevant to policy makers. Therefore, the development of a multi-criteria approach to priority setting is necessary, and this has indeed recently been identified as one of the most important issues in health system research. In other scientific disciplines, multi-criteria decision analysis is well developed, has gained widespread acceptance and is routinely used. This paper presents the main principles of multi-criteria decision analysis. There are only a very few applications to guide resource allocation decisions in health. We call for a shift away from present priority setting tools in health--that tend to focus on single criteria--towards transparent and systematic approaches that take into account all relevant criteria simultaneously.
    背景与目标: :卫生干预措施的优先级设置通常是临时的,资源没有得到最佳利用。潜在的问题是多个标准起着作用,并且决策很复杂。可以选择干预措施,以最大程度地提高总体人口健康,减少弱势或弱势群体的健康不平等,广告/或应对危及生命的情况,所有这些都涉及实际和预算方面的限制。这是决策者通常无助于理性解决的典型问题。他们倾向于使用启发式或直观的方法来简化复杂性,并且在此过程中,重要的信息将被忽略。接下来,政策制定者可能只出于政治动机选择干预措施。这表明需要采取合理和透明的方法来确定优先事项。在过去的几十年中,已经开发出许多方法,包括循证医学,疾病负担分析,成本效益分析和公平性分析。但是,这些方法仅集中于单个标准,而实际上,决策者需要在选择时同时考虑多个标准。此外,它们并未涵盖与决策者相关的所有标准。因此,有必要开发一种确定优先级的多标准方法,并且最近确实将其确定为卫生系统研究中最重要的问题之一。在其他科学学科中,多准则决策分析已经得到了很好的发展,已经得到了广泛的认可并被常规使用。本文介绍了多准则决策分析的主要原理。只有很少的应用程序可以指导健康状况中的资源分配决策。我们呼吁从目前卫生领域的优先重点设定工具(倾向于只关注单一标准)转变为同时考虑所有相关标准的透明,系统的方法。
  • 【FOXP3的分析揭示了其作为转录阻遏物所需的多个结构域。】 复制标题 收藏 收藏
    DOI:10.4049/jimmunol.177.5.3133 复制DOI
    作者列表:Lopes JE,Torgerson TR,Schubert LA,Anover SD,Ocheltree EL,Ochs HD,Ziegler SF
    BACKGROUND & AIMS: :Foxp3 has been shown to be both necessary and sufficient for the development and function of naturally arising CD4+ CD25+ regulatory T cells in mice. Mutation of Foxp3 in Scurfy mice and FOXP3 in humans with IPEX results in fatal, early onset autoimmune disease and demonstrates the critical role of FOXP3 in maintaining immune homeostasis. The FOXP3 protein encodes several functional domains, including a C2H2 zinc finger, a leucine zipper, and a winged-helix/forkhead (FKH) domain. We have shown previously that FOXP3 functions as a transcriptional repressor and inhibits activation-induced IL-2 gene transcription. To characterize the role of each predicted functional domain on the in vivo activity of FOXP3, we have evaluated the location of point mutations identified in a large cohort of patients with the immune dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome (IPEX) and found them to cluster primarily within the FKH domain and the leucine zipper, but also present within the poorly defined N-terminal portion of the protein. The molecular functions of each of the IPEX-targeted domains were investigated. We show that FOXP3 is constitutively localized to the nucleus and this localization requires sequences at both the amino and C-terminal ends of its FKH domain. Moreover, FOXP3 was found to homodimerize through its leucine zipper. We also identify a novel functional domain within the N-terminal half of FOXP3, which is required for FOXP3-mediated repression of transcription from both a constitutively active and a NF-AT-inducible promoter. Furthermore, we demonstrate that IPEX mutations in these domains correlate with deficiencies in FOXP3 repressor function, corroborating their in vivo relevance.
    背景与目标: 已经证明:Foxp3对于小鼠中天然产生的CD4 CD25调节性T细胞的发育和功能既必要又充分。用IPEX突变的Scurfy小鼠中的Foxp3和人类中的FOXP3突变会导致致命的早期发作的自身免疫性疾病,并证明FOXP3在维持免疫稳态方面的关键作用。 FOXP3蛋白编码几个功能域,包括C2H2锌指,亮氨酸拉链和带翼螺旋/叉头(FKH)域。先前我们已经证明FOXP3充当转录阻遏物,并抑制激活诱导的IL-2基因转录。为了表征每个预测功能结构域对FOXP3体内活性的作用,我们评估了在一大批患有免疫功能异常,多内分泌病,肠病,X连锁综合征(IPEX)的患者中鉴定出的点突变的位置,并发现它们主要聚集在FKH结构域和亮氨酸拉链中,但也存在于蛋白质的N末端定义不明确的区域。研究了每个IPEX靶向域的分子功能。我们显示FOXP3组成性地定位于原子核,并且此定位需要在其FKH域的氨基和C末端都具有序列。此外,发现FOXP3通过其亮氨酸拉链同源二聚体。我们还确定了FOXP3 N末端一半内的新型功能域,这是FOXP3介导的从组成型活性启动子和NF-AT诱导型启动子转录抑制所必需的。此外,我们证明这些域中的IPEX突变与FOXP3阻遏物功能的缺陷相关,从而证实了它们的体内相关性。
  • 【伴刀豆球蛋白A,但不是糖化白蛋白,可增加体外血管性假血友病因子的内皮下沉积。】 复制标题 收藏 收藏
    DOI:10.1080/10623320600903916 复制DOI
    作者列表:Nizheradze K
    BACKGROUND & AIMS: :Diabetes is associated with augmentation of prothrombogenic von Willebrand factor (vWF) content in plasma. Earlier, the author and colleagues have shown that high glucose and insulin do not appreciably influence deposition of vWF into the subendothelial extracellular matrix (SECM) produced by cultured human umbilical vein endothelial cells (HUVECs). In the present work, the author used this model to test the effects of nonenzymatically glycated albumin (Glyc-HSA) and two lectins, concanavalin A (ConA) and wheat germ agglutinin (WGA), on vWF deposition into the SECM. First-passage HUVECs were seeded into gelatin-coated 96-well plates and cultured for 6 to 7 days. HSA or Glyc-HSA (at concentrations 25, 50, and 100 microg/mL), and WGA or ConA (4, 8, and 16 microg/mL) were added 3 h after seeding. Cell viability was tested by the MTT method. To determine vWF contents in the SECM, HUVECs were detached by treatment with NH4OH and the residual material was used as a solid phase in an enzyme-linked immunosorbent assay (ELISA)-like assay with primary (anti-vWF) and secondary (peroxidase-conjugated) antibodies. Addition of Glyc-HSA did not essentially influence VWF contents in the SECM (A490 was 0.226 versus 0.268 at 0 and 100 microg/mL, respectively; p > .05, n = 16). Cultivation in the presence of WGA led to the deterioration of cell viability, which was accompanied by a significant decrease of vWF in the SECM (0.248 versus 0.128 at 0 and 16 microg/mL, respectively; p < .001, n = 16). ConA did not influence viability of HUVECs, but this lectin at all concentrations consistently increased the deposition of vWF (up to 164% relative to control, p <.001; n = 16). These data indicate that endothelial carbohydrate determinants and corresponding ligands (namely, mannose-specific lectins) may be involved in the regulation of production and deposition of vWF.
    背景与目标: 糖尿病与血浆中促血栓性血管性血友病因子(vWF)含量的增加有关。早前,作者和同事已经表明,高葡萄糖和胰岛素不会明显影响vWF在培养的人脐静脉内皮细胞(HUVEC)产生的内皮下细胞外基质(SECM)中的沉积。在目前的工作中,作者使用该模型测试了非酶促糖基化白蛋白(Glyc-HSA)和两种凝集素,伴刀豆球蛋白A(ConA)和小麦胚芽凝集素(WGA)对vWF沉积到SECM中的影响。将第一代HUVEC接种到明胶包被的96孔板中,并培养6至7天。播种后3小时,添加HSA或Glyc-HSA(浓度分别为25、50和100微克/毫升)和WGA或ConA(4、8和16微克/毫升)。细胞存活力通过MTT方法测试。为了确定SECM中的vWF含量,通过用NH4OH处理将HUVEC分离,并将残留的物质作为固相用于类似酶联免疫吸附测定(ELISA)的类似测定中,该测定具有一级(抗vWF)和二级(过氧化物酶-偶联的)抗体。添加Glyc-HSA基本上不影响SECM中的VWF含量(0和100 microg / mL时A490分别为0.226和0.268; p> .05,n = 16)。在存在WGA的情况下进行培养会导致细胞活力下降,并伴有SECM中的vWF显着下降(0和16 microg / mL分别为0.248和0.128; p <.001,n = 16)。 ConA不会影响HUVEC的生存力,但是该凝集素在所有浓度下均会持续增加vWF的沉积(相对于对照,高达164%,p <.001; n = 16)。这些数据表明内皮糖决定簇和相应的配体(即,甘露糖特异性凝集素)可能参与vWF的产生和沉积的调节。
  • 【注意缺陷多动障碍可能与中枢性脑源性神经营养因子活性降低有关:临床和治疗意义。】 复制标题 收藏 收藏
    DOI:10.1016/j.mehy.2006.06.025 复制DOI
    作者列表:Tsai SJ
    BACKGROUND & AIMS: :Attention-deficit hyperactivity disorder (ADHD) is a common childhood psychiatric disorder. Despite intensive research efforts, the aetiology of ADHD remains unknown. Current evidence suggests that the aetiology of ADHD is heterogeneous, comprising of multiple factors. Recently, it has been proposed that brain-derived neurotrophic factor (BDNF), a member of the neurotrophic factor family, may be implicated in the pathogenesis of ADHD. This hypothesis is supported by recent genetic studies in ADHD. Drawing on findings from studies into the drugs for ADHD relating to central BDNF expression, hyperactivity in BDNF knockout mice, BDNF effects in midbrain dopaminergic function and the close association between BDNF and the dopamine transporter (an important molecule for ADHD pathogenesis), it is proposed here that decreased central BDNF, particularly in the midbrain region, may play an important role in the pathogenesis ADHD. This hypothesis may have some implications for clinical findings in ADHD (for example, the co-morbidity between ADHD and major depression), and provide a new direction for the development of medication for ADHD treatment.
    背景与目标: 注意缺陷多动障碍(ADHD)是儿童期常见的精神病。尽管进行了深入的研究,但多动症的病因仍然未知。当前证据表明,ADHD的病因是异质的,由多种因素组成。最近,已经提出,脑源性神经营养因子(BDNF),神经营养因子家族的成员,可能与ADHD的发病有关。这一假说得到了多动症最近的遗传学研究的支持。根据对ADHD药物的研究发现,该药物与中枢BDNF表达,BDNF基因敲除小鼠的过度活跃,BDNF对中脑多巴胺能功能的影响以及BDNF与多巴胺转运蛋白(ADHD发病机理的重要分子)之间的紧密联系有关,因此提出了这一建议。在这里,中央BDNF的降低,特别是在中脑区域,可能在ADHD的发病中起重要作用。该假设可能对ADHD的临床发现有一定的影响(例如,ADHD与严重抑郁症的合并症),并为ADHD治疗药物的开发提供了新的方向。

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