BACKGROUND & AIMS: A 38-year-old man, blood group A+, was allotransplanted for multiple myeloma from his fully matched sister, blood group O+. Anti-A antibodies IgG and IgM titres of the donor were low. Allogeneic peripheral blood stem cells were harvested by leukapheresis after subcutaneous administration of G-CSF. Rapid engraftment occurred since 5.6 x 10(9)/l leukocytes were achieved on day +9 post-transplant. At this time a severe immune haemolytic syndrome occurred and direct antiglobulin test was positive (IgG and C3d). Elution showed an anti-A specificity. Evolution was rapidly unfavourable related to multiorgan failure. The patient died on day +20 post-transplant.

背景与目标： 一名38岁的A血型男子从其完全匹配的姐姐O血型中同种异体移植了多发性骨髓瘤。供体的抗A抗体IgG和IgM滴度较低。皮下给予g-csf后，通过白细胞分离术收集异基因外周血干细胞。由于在移植后第9天实现了5.6 × 10(9)/l白细胞，因此发生了快速植入。此时发生了严重的免疫溶血综合征，直接抗球蛋白试验呈阳性 (IgG和C3d)。洗脱显示出抗A特异性。进化与多器官衰竭迅速不利。患者在移植后第20天死亡。

BACKGROUND & AIMS: :How neutrophils (polymorphonuclear neutrophils, PMNs) damage vessels in leukocytoclastic vasculitis (LcV) mediated by immune complexes (ICs) is unclear. If degradative enzymes and oxygen radicals are released from PMNs while adhering to the inner side of the vessel wall, they could be washed away by the blood stream or neutralized by serum protease inhibitors. We investigated if in LcV PMNs could damage vessels from the tissue side after transmigration. We used CD18-deficient (CD18-/-) mice because the absence of CD18 excludes transmigration of PMNs. When eliciting the Arthus reaction in ears of CD18-/- mice, deposition of ICs was not sufficient to recruit PMNs or to induce IC-mediated LcV. Injection of PMNs intradermally in CD18-/- mice allowed us to investigate if bypassing diapedesis and placing PMNs exclusively on the abluminal side leads to vascular destruction. We found that injected PMNs gathered around perivascular ICs, but did not cause vessel damage. Only intravenous injection of wild-type PMNs could re-establish the Arthus reaction in CD18-/- mice. Thus, PMNs cause vessel damage during diapedesis from the luminal side, but not from the perivascular space. We suggest that in order to shield the cytotoxic products from the blood stream, ICs induce particularly tight interactions between them, PMNs and endothelial cells.

背景与目标： : 中性粒细胞 (多形核中性粒细胞，PMNs) 如何损害由免疫复合物 (ICs) 介导的白细胞碎裂血管炎 (LcV) 中的血管尚不清楚。如果降解酶和氧自由基从pmn中释放出来，同时粘附在血管壁的内侧，则它们可能会被血流冲走或被血清蛋白酶抑制剂中和。我们调查了在LcV中，pmn是否会在迁移后从组织侧损坏血管。我们使用CD18-deficient (CD18-/-) 小鼠，因为缺少CD18排除了pmn的迁移。当在CD18-/-小鼠的耳朵中引起Arthus反应时，ICs的沉积不足以募集pmn或诱导IC介导的LcV。在CD18-/-小鼠中皮内注射pmn使我们能够研究是否绕过血管造影并仅将pmn放置在管腔侧会导致血管破坏。我们发现注射的pmn聚集在血管周围ICs周围，但不会引起血管损伤。只有静脉注射野生型PMNs才能在CD18-/-小鼠中重新建立Arthus反应。因此，pmn在从管腔侧 (而不是从血管周围空间) 进行的排尿过程中会引起血管损伤。我们建议，为了从血流中屏蔽细胞毒性产物，ICs在它们，pmn和内皮细胞之间诱导特别紧密的相互作用。

BACKGROUND & AIMS: AIM:To investigate the dynamic alteration of telomerase expression during development of hepatocellular carcinoma (HCC) and its diagnostic implications in liver tissues or peripheral blood mononuclear cells for HCC. METHODS:Dynamic expressions of liver telomerase during malignant transformation of hepatocytes were observed in Sprague-Dawly (SD) rats fed with 0.05% of 2-fluoenyacetamide (2-FAA). Total RNA and telomerase were extracted from rat or human liver tissues. The telomerase activities in livers and in circulating blood were detected by a telomeric repeat amplification protocol-enzyme-linked immunosorbent assay (TRAP-ELISA), and its diagnostic value was investigated in patients with benign or malignant liver diseases. RESULTS:The hepatoma model displayed the dynamic expression of hepatic telomerase during HCC development. The telomerase activities were consistent with liver total RNA levels (r = 0.83, P<0.01) at the stages of degeneration, precancerosis, and cancerization of hepatocytes. In HCC patients, the telomerase levels in HCC tissues were significantly higher than in their adjacent non-cancerous tissues, but liver total RNA levels were lower in the former than in the latter. Although the circulating telomerase of HCC patients was abnormally expressed among patients with chronic liver diseases, the telomerase activity was a non-specific marker for HCC diagnosis, because the incidence was 15.7% in normal control, 25% in chronic hepatitis, 45.9% in liver cirrhosis, and 85.2% in HCC, respectively when absorbance value of telomerase activity was more than 0.2. If the value was over 0.6, the incidence was 60% in HCC group and 0% in any of the others (P<0.01) except in two cases with liver cirrhosis. However, the combination of circulating telomerase with serum alpha-fetoprotein level could increase the positive rate and the accuracy (92.6%, 125 of 135) of HCC diagnosis. CONCLUSION:The overexpression of telomerase is associated with HCC development, and its abnormality in liver tissues or in peripheral blood could be a useful marker for diagnosis and prognosis of HCC.

背景与目标：

BACKGROUND & AIMS: :A photoplethysmographic (PPG) technique to assess blood flow in bone tissue has been developed and tested. The signal detected by the PPG consists of a constant-level (DC) component-which is related to the relative vascularization of the tissue-and a pulsatile (AC) component-which is synchronous with the pumping action of the heart. The PPG probe was applied on the skin over the patella. The probe uses near-infrared (804 nm) and green (560 nm) light sources and the AC component of the PPG signals of the two wavelengths was used to monitor pulsatile blood flow in the patellar bone and the overlying skin, respectively. Twenty healthy subjects were studied and arterial occlusion resulted in elimination of PPG signals at both wavelengths, whereas occlusion of skin blood flow by local surface pressure eliminated only the PPG signal at 560 nm. In a parallel study on a physical model with a rigid tube we showed that the AC component of the PPG signal originates from pulsations of blood flow in a rigid structure and not necessarily from volume pulsations. We conclude that pulsatile blood flow in the patellar bone can be assessed with the present PPG technique.

背景与目标： : 已经开发并测试了一种用于评估骨组织血流的光电容积描记 (PPG) 技术。PPG检测到的信号由恒定水平 (DC) 分量 (与组织的相对血管化有关) 和脉动 (AC) 分量组成，该分量与心脏的泵送动作同步。将PPG探针涂在髌骨上的皮肤上。探针使用近红外 (804 nm) 和绿色 (560 nm) 光源，并且两个波长的PPG信号的AC分量分别用于监测髌骨和上覆皮肤中的脉动血流。研究了20名健康受试者，动脉闭塞导致消除了两个波长处的PPG信号，而局部表面压力对皮肤血流的闭塞仅消除了560 nm处的PPG信号。在对具有刚性管的物理模型的并行研究中，我们表明PPG信号的AC分量来自刚性结构中的血流脉动，而不一定来自体积脉动。我们得出的结论是，可以使用目前的PPG技术评估pa骨中的脉动血流。

BACKGROUND & AIMS: :There is an abundance of clinical applications using human umbilical cord blood (HUCB) as a source for stem cell populations. Other than haematopoietic progenitors, there are mesenchymal, endothelial stem cells and neuronal precursors, in varying quantities, that are found in human umbilical cord blood. These may be useful in diseases such as immune deficiency and autoimmune disorders. Considering issues of safety, availability, transplant methodology, rejection and side effects, it is contended that a therapeutic stem cell transplant, utilizing stem cells from HUCB, provides a reliable repository of early precursor cells that can be useful in a great number of diverse conditions. Drawbacks of relatively smaller quantities of mononucleated cells in one unit of cord blood can be mitigated by in-vitro expansion procedures, improved in-vivo signalling, and augmentation of the cellular milieu, while simultaneously choosing the appropriate transplantation site and technique for introduction of the stem cell graft.

背景与目标： : 使用人脐带血 (HUCB) 作为干细胞种群的来源有很多临床应用。除造血祖细胞外，在人脐带血中还发现了各种数量的间充质，内皮干细胞和神经元前体。这些可能在诸如免疫缺陷和自身免疫性疾病等疾病中有用。考虑到安全性，可用性，移植方法，排斥反应和副作用等问题，有人认为，利用HUCB干细胞的治疗性干细胞移植提供了可靠的早期前体细胞储存库，可用于多种多样的条件。可以通过体外扩增程序，改善体内信号传导和增加细胞环境，同时选择合适的移植部位和技术来引入脐带血，减轻脐带血中相对较少数量的单核细胞的缺点。干细胞移植物。

BACKGROUND & AIMS: This study was designed to validate the utility of a commercial portable clinical blood analyzer (PCBA) in ground-based studies and on the space shuttle. Ionized calcium, pH, electrolytes, glucose, and hematocrit were determined. Results agreed well with those from traditional laboratory methods, and the PCBA demonstrated good between-day precision for all analytes. In-flight analysis of control samples revealed differences in one analyte (sodium). There were few changes in crew members' results during flight, and these were expected. Potassium increased in flight compared with before flight, and potassium, pH, and hematocrit decreased after flight. Ionized calcium was decreased in flight and on landing day. Changes during flight were likely related to sample collection technique. Postflight changes likely reflected the fluid redistribution that occurs after exposure to weightlessness. These data confirm that the PCBA is a reliable instrument for most analytes, and can provide important medical data in remote locations, such as orbiting spacecraft.

背景与目标： 这项研究旨在验证商用便携式临床血液分析仪 (PCBA) 在地面研究和航天飞机上的实用性。测定了离子钙，pH，电解质，葡萄糖和血细胞比容。结果与传统实验室方法的结果非常吻合，并且PCBA对所有分析物均显示出良好的日间精度。对照样品的飞行分析显示一种分析物 (钠) 存在差异。在飞行过程中，机组人员的结果几乎没有变化，这是可以预料的。与飞行前相比，飞行中钾增加，飞行后钾，pH和血细胞比容降低。在飞行中和着陆日，离子钙减少。飞行过程中的变化可能与样品收集技术有关。飞行后的变化可能反映了失重后发生的流体重新分布。这些数据证实了PCBA对于大多数分析物来说是一种可靠的仪器，并且可以在偏远的地方提供重要的医学数据，例如绕行航天器。

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：

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 (CMV) 病毒载量与多形核白细胞 (PMNL) 和血浆样本中特定UL97突变之间的时间关系，该患者患有更昔洛韦耐药的CMV视网膜炎。
方法 : 使用非分子方法和定量聚合酶链反应 (PCR) 分析方法分析了连续的PMNL和血浆样品，以确定CMV病毒载量。使用巢式PCR和限制性内切酶分析对相同样品进行了更昔洛韦抗性最常见突变的筛选。
结果 : 在CMV视网膜炎进展时 (更昔洛韦6个月后)，在PMNL和血浆样品中发现CMV DNA载量迅速增加。这种增加与同一样品中出现特定突变 (V594) 和更昔洛韦耐药血液分离株的回收平行。但是，在该患者中，唯一可以基本预测CMV疾病进展的测试是使用PMNL的定量PCR测定，以及在较小程度上使用pp65抗原血症测定。
结论 : 使用敏感的检测方法 (例如PCR) 定量评估PMNL中的CMV病毒载量似乎是监测艾滋病患者抗病毒治疗的有希望的方法。此外，可以直接在PMNL和血浆样品中检测到赋予更昔洛韦抗性的常见突变。

BACKGROUND & AIMS: UNLABELLED:In this study, we assessed the accuracy and reliability of MRI-guided SPECT measurements of medial temporal lobe blood flow in Alzheimer's disease (AD).

METHODS:Interactively aligned three-dimensional MP-RAGE MRI and 99mTc-HMPAO SPECT images were used for MRI-guided measurement of medial temporal lobe CBF in eight control subjects and eight patients with probable AD. Intraoperator reliability was assessed by repeated alignment and measurement by one experienced operator. Accuracy was assessed by examining two subjects with fiducial markers.

RESULTS:The alignment error was less than 1 SPECT pixel size (3.5 mm) and the coefficient of variation in repeated measures of medial temporal-to-cerebellar CBF ratios was 3.2%. The difference in mean medial temporal-to-cerebellar CBF ratios between eight control subjects and eight AD patients was 12%. Also by using three-dimensional seed-grow defined healthy brain reference regions, there were significant differences between control subjects and AD patients in medial temporal blood flow. Furthermore, the volumes of the MRI-defined medial temporal ROIs were smaller in the AD patients. The best separation between AD patients and control subjects was achieved by combining MRI measurements of atrophy and SPECT measurements of CBF.

CONCLUSION:These data show that the accuracy and reliability of MRI-guided SPECT measurements of medial temporal CBF clearly allow the detection of changes in AD. Also, a direct comparison of structural and functional changes is possible by this methodology, which might improve the early diagnosis of AD.

背景与目标： 未标记 : 在这项研究中，我们评估了MRI引导的SPECT测量阿尔茨海默氏病 (AD) 内侧颞叶血流的准确性和可靠性。
方法 : 交互式对齐的三维MP-RAGE MRI和99mTc-HMPAO SPECT图像用于MRI引导的8名对照受试者和8名可能患有AD的患者的内侧颞叶CBF测量。一位经验丰富的操作员通过重复对准和测量来评估操作员内部的可靠性。通过检查两个具有基准标记的受试者来评估准确性。
结果 : 对齐误差小于1 SPECT像素大小 (3.5毫米)，并且重复测量的变异系数内侧颞-小脑CBF比3.2%。12% 了八名对照受试者和八名AD患者之间平均内侧颞与小脑CBF比率的差异。同样，通过使用三维种子生长定义的健康大脑参考区域，对照组和AD患者在内侧颞血流方面存在显着差异。此外，在AD患者中，MRI定义的内侧颞roi的体积较小。通过结合萎缩的MRI测量和CBF的SPECT测量，可以实现AD患者与对照组之间的最佳分离。
结论 : 这些数据表明，MRI引导的内侧颞CBF SPECT测量的准确性和可靠性显然可以检测AD的变化。此外，通过这种方法可以直接比较结构和功能变化，这可能会改善AD的早期诊断。

BACKGROUND & AIMS: OBJECTIVE:To assess and compare immediate effects of chest physiotherapy with positive expiratory pressure (PEP) versus oscillating PEP on transcutaneously measured blood-gas tensions in patients with cystic fibrosis. METHODS:Fifteen patients (mean age 12.5 y, range 6.9-21.5 y) participated. The treatments were randomized and performed on 2 separate occasions, 8 weeks apart. Spirometry was conducted before and after each treatment. We transcutaneously measured oxygen tension (P(tO2). RESULTS:There were no changes in spirometry values. During PEP, different trends in blood-gas tension were seen, and there were no consistent changes. During oscillating PEP, P(tO2) increased and P(tCO2) decreased. During oscillating PEP, P(tCO2) was lower and the intra-individual change in P(tCO2) was more pronounced than during PEP. The results obtained immediately after oscillating PEP showed a higher P(tO2) and a lower P(tCO2) than with PEP. CONCLUSION:PEP and oscillating PEP can both cause transitory effects on blood gases in patients with cystic fibrosis. However, oscillating PEP alters blood-gas tensions more than does PEP, and hyperventilation during oscillating PEP may reduce treatment time.

背景与目标：

BACKGROUND & AIMS: Activated neutrophils take a long time to pass through a narrow lumen like a micropore, and are supposed to play a deteriorating effect on microcirculation. Although the activation of neutrophils has been demonstrated in Behçet's disease, nobody analyzes the clinical activity of the disease by means of the rheological measure of neutrophils activity. Using a micropore (pore diameter 5 microns) filtration technique, we measured the filtration time of peripheral blood neutrophils, as a rheological measure of their activity, in order to determine the clinical activity of Behçet's disease. Twenty-one patients with Behçet's disease and 14 healthy control individuals were enrolled in the study. Symptoms and signs exhibited in the patients led us to distinguish the Behçet's disease into inactive and active cases. The latter were further differentiated into cases with absent symptoms and with present symptoms. Neutrophil filtration times were 11.5 +/- 4.8 s in the active cases with present symptoms, which were significantly (P < 0.05) larger than those (7.4 +/- 1.9 s) in the active cases with absent symptoms. The latter filtration times were further significantly (P < 0.001) larger than values (3.7 +/- 1.3 s) in the inactive cases and also those (4.8 +/- 1.2 s) in control subjects. Furthermore, increases in the filtration time obtained immediately after the exposure of cells to the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (FMLP10 nM) were significantly (P < 0.01) larger in the active cases with present symptoms than those in the active cases with absent symptoms. The latter were also larger, but not significantly, than those in the inactive cases, and were significantly (P < 0.01) larger than those in control subjects. The present results demonstrate that the micropore filtration method reflects well the rheological activity of neutrophils as well as the clinical status of Behçet's disease. This method is much better than the measurement of O2 production to differentiate between active cases with absent symptoms and inactive patients or even control individuals. Furthermore, it is more sensitive and useful than laboratory data like the CRP value or the number of peripheral blood neutrophils.

背景与目标： 活化的中性粒细胞需要很长时间才能通过像微孔一样的狭窄管腔，并且应该对微循环起到恶化的作用。尽管在beh ç et病中已经证明了中性粒细胞的激活，但没有人通过中性粒细胞活性的流变学测量来分析该疾病的临床活性。使用微孔 (孔径5微米) 过滤技术，我们测量了外周血中性粒细胞的过滤时间，作为其活性的流变学指标，以确定白塞氏病的临床活性。21名beh ç et病患者和14名健康对照者参加了这项研究。患者表现出的症状和体征使我们将白塞氏病区分为不活跃和活跃的病例。后者进一步分为无症状和现有症状的病例。有症状的活动病例的中性粒细胞过滤时间为11.5 +/- 4.8 s，显著 (P < 0.05) 大于无症状的活动病例的中性粒细胞过滤时间 (7.4 +/- 1.9 s)。后者的过滤时间进一步显著 (P < 0.001) 大于非活性情况下的值 (3.7 +/- 1.3 s)，也大于对照受试者中的值 (4.8 +/- 1.2 s)。此外，在细胞暴露于趋化肽甲酰基-甲硫酰基-亮氨酸-苯丙氨酸 (fmlp10nm) 后立即获得的过滤时间的增加在存在症状的活动病例中比在不存在症状的活动病例中显着 (P < 0.01) 大。后者也比不活跃的情况更大，但不显著，并且显著 (P < 0.01) 大于对照组。目前的结果表明，微孔过滤方法很好地反映了嗜中性粒细胞的流变活性以及白塞病的临床状况。此方法比O2产生的测量要好得多，可以区分无症状的活跃病例和不活跃的患者甚至对照组。此外，它比CRP值或外周血中性粒细胞数量等实验室数据更敏感和有用。

BACKGROUND & AIMS: Acute normovolemic hemodilution has been reported to result in blood savings varying from 18% to 90%. Very few of these are randomized prospective studies. This study attempts to determine the blood transfusion savings if acute normovolemic hemodilution is used in combination with autologous predonated blood and cell saver. Thirty-three patients undergoing total hip arthroplasty were assigned randomly to one of two groups (control, n = 16; hemodilution, n = 17). Patients in both groups entered an autologous predonation program if cleared medically and were placed on Cell Saver intraoperatively and in the postanesthesia care unit. In addition, the hemodilution group underwent acute normovolemic hemodilution preoperatively. Only 41% of the patients in the hemodilution group required any autologous blood transfusion as compared with 75% of the control group. In addition, the hemodilution group required a mean lower quantity of autologous blood transfusion (41% of the estimated blood loss) as compared with the control group (71%). The net anesthesia time increased by an average of 11.4 minutes in the hemodilution group. Acute normovolemic hemodilution is a safe procedure even in an older patient population. Hemodilution resulted in fewer patients needing autologous predonated blood transfusions. The major benefit of hemodilution was seen when predonation was not possible.

背景与目标： 据报道，急性等容量血液稀释会导致血液节约从18% 到90% 不等。其中很少是随机前瞻性研究。这项研究试图确定如果将急性等容量血液稀释与自体预献血和细胞保护剂结合使用，则可以节省输血。将接受全髋关节置换术的33例患者随机分为两组之一 (对照组，n = 16; 血液稀释，n = 17)。如果经过医学批准，两组患者都进入了自体预捐赠计划，并在术中和麻醉后护理病房中放置在Cell Saver上。此外，血液稀释组术前进行了急性等容量血液稀释。与对照组的75% 相比，血液稀释组中只有41% 的患者需要任何自体输血。此外，与对照组 (71%) 相比，血液稀释组需要平均较低量的自体血回输 (41% 估计的失血量)。血液稀释组的净麻醉时间平均增加11.4分钟。即使在老年患者中，急性等容积量血液稀释也是一种安全的方法。血液稀释导致需要自体预输血的患者减少。当无法进行预捐赠时，可以看到血液稀释的主要好处。

BACKGROUND & AIMS: The circumventricular organs (CVOs) in the brain are without a blood-brain barrier (BBB) and as such directly exposed to blood plasma constituents and blood-borne pathogens. In light of previous studies showing discrepancies regarding the immunocompetence of these organs, we initiated the present study to provide a comprehensive immunohistochemical analysis of the cellular expression of immune-associated antigens within the pineal gland, area postrema and the subfornical organ. In all CVOs, subpopulations of cells morphologically similar to complement receptor type 3 immunoreactive microglial/macrophage cells expressed major histocompatibility complex (MHC) class II antigen, leucocyte common antigen (LCA/CD45), as well as CD4 and ED1 antigen. Based on morphological criteria the MHC class II antigen expressing cells could be grouped into a major population of classical parenchymal and perivascular ramified microglial cells and a minor population presenting itself as scattered or small groups of rounded macrophage-like cells. CD4 and ED1 antigen were expressed by both cell types. CD45 was preferentially expressed by macrophage-like cells. MHC class I antigen was expressed by the vascular endothelium in both BBB-protected and BBB-deficient areas and was additionally present as a lattice-like network throughout the BBB-deficient parenchyma in all CVOs. The results suggest that the BBB-free areas of the brain besides being constantly surveyed by blood-borne macrophages, possess an intrinsic immune surveillance system based on resting and activated microglial cells, which may function as a non-endothelial, cellular barrier against blood-borne pathogens.

背景与目标： 大脑中的心室器官 (CVOs) 没有血脑屏障 (BBB)，因此直接暴露于血浆成分和血液传播的病原体。鉴于先前的研究显示这些器官的免疫能力存在差异，我们启动了本研究，以提供松果体，后区域和下器官内免疫相关抗原的细胞表达的全面免疫组织化学分析。在所有cvo中，形态上与补体受体3型免疫反应性小胶质细胞/巨噬细胞相似的细胞亚群表达主要组织相容性复合物 (MHC) II类抗原，白细胞共同抗原 (LCA/CD45) 以及CD4和ED1抗原。根据形态学标准，可以将MHC II类抗原表达细胞分为主要的经典实质和血管周围分支的小胶质细胞群，以及少量的呈分散或小群的圆形巨噬细胞样细胞群。两种细胞类型均表达CD4和ED1抗原。CD45优先由巨噬细胞样细胞表达。MHC I类抗原在BBB保护和BBB缺陷区域均由血管内皮表达，并且在所有cvo的BBB缺陷实质中还以晶格状网络的形式存在。结果表明，大脑的无BBB区域除了不断受到血源性巨噬细胞的调查外，还具有基于静息和活化的小胶质细胞的内在免疫监视系统，该系统可能是针对血源性病原体的非内皮细胞屏障。

BACKGROUND & AIMS: :Wild-type (WT) sequence p53 peptides are attractive candidates for broadly applicable cancer vaccines. The aim of this study was to evaluate the potential of a WT p53-based immunotherapeutic approach for patients with hepatocellular carcinoma (HCC). Circulating CD8+ T cells specific for WT p53(149-157) and WT p53(264-272) HLA-A*0201 restricted epitopes were directly identified in the peripheral blood by the use of peptide/HLA-A2.1 tetramers in 24 HCC patients. Cytotoxic T lymphocyte (CTL) activity after WT p53 peptide-specific stimulation was assessed by analysis of granzyme B and interferon-gamma mRNA transcription, using a quantitative real-time polymerase chain reaction assay. Tumor immunophenotyping was performed to evaluate the p53 status, the expression of major histocompatibility complex (MHC) and costimulatory molecules in freshly isolated tumor cells. HCC patients exhibited significantly higher frequencies of WT p53-specific memory CD8+ T cells and stronger WT p53-specific CTL activity, when compared with healthy controls. Increased frequencies of p53-specific CD8+ T cells and their activity correlated with selective HLA-A2 allele loss and reduced costimulatory molecule expression of tumor cells. Moreover, augmented numbers of p53-specific T cells coincided with high MHC class II expression in tumor cells but were inversely related to the T status of the tumor node metastasis staging system. Our results indicate the existence of natural immunosurveillance and tumor immune evasion, involving a T cell response against WT p53 tumor antigen in patients with HCC. These findings may have important implications for the future development of cancer vaccines.

背景与目标： : 野生型 (WT) 序列p53肽是广泛适用的癌症疫苗的有吸引力的候选者。这项研究的目的是评估WT p53-based免疫治疗方法对肝细胞癌 (HCC) 患者的潜力。通过在24例HCC患者中使用肽/HLA-A2.1四聚体，在外周血中直接鉴定了对WT p53(149-157) 和WT p53(264-272) hla-a * 0201限制性表位特异性的循环CD8 + T细胞。使用定量实时聚合酶链反应测定法，通过分析颗粒酶B和干扰素-γ mRNA转录来评估WT p53肽特异性刺激后的细胞毒性T淋巴细胞 (CTL) 活性。进行肿瘤免疫表型分析以评估新鲜分离的肿瘤细胞中的p53状态，主要组织相容性复合物 (MHC) 和共刺激分子的表达。与健康对照组相比，HCC患者表现出明显更高的WT p53-specific记忆CD8 + T细胞频率和更强的WT p53-specific CTL活性。p53-specific CD8 + T细胞的频率增加及其活性与肿瘤细胞的选择性HLA-A2等位基因丢失和共刺激分子表达降低相关。此外，p53-specific T细胞数量的增加与肿瘤细胞中高MHC II类表达相吻合，但与肿瘤淋巴结转移分期系统的T状态成反比。我们的结果表明存在自然免疫监视和肿瘤免疫逃避，涉及HCC患者针对WT p53肿瘤抗原的T细胞反应。这些发现可能对癌症疫苗的未来发展具有重要意义。

BACKGROUND & AIMS: :This study was designed to evaluate the hypothesis that administration of a replication-deficient, recombinant adenovirus vector to the epithelial surface of the respiratory tract can be used to deliver a recombinant protein to the systemic circulation in sufficient quantities to evoke a systemic response appropriate to the recombinant protein. We administered AdCMV.TPO-an adenovirus vector containing an expression cassette coding for the human thrombopoietin (TPO) cDNA-to the respiratory epithelium of immunocompetent Balb/c mice. Over the following week, serum human TPO levels were elevated, platelet levels increased more than sixfold, and megakaryocytosis was evident in bone marrow. This strategy may be a useful approach to the nonparenteral administration of a variety of therapeutic recombinant proteins, such as those relevant to clotting, endocrine function, and bone-marrow function.

背景与目标： : 这项研究旨在评估以下假设: 将复制缺陷型重组腺病毒载体施用到呼吸道上皮表面可用于将重组蛋白以足够量递送到体循环，以引起适合的全身反应。重组蛋白。我们向具有免疫活性的Balb/c小鼠的呼吸道上皮施用AdCMV.TPO-一种含有编码人血小板生成素 (TPO) cDNA的表达盒的腺病毒载体。在接下来的一周中，血清人TPO水平升高，血小板水平增加了六倍以上，并且骨髓中巨核细胞增多。该策略可能是非胃肠外给药多种治疗性重组蛋白的有用方法，例如与凝血，内分泌功能和骨髓功能有关的蛋白。