Follicular dendritic cell accessory activity crosses MHC and species barriers

Productive follicular dendritic cell (FDC)-B cell interactions appear to involve critical ligand-receptor interactions. Immune complexes (IC) on FDC activate complement and provide FDC with a complement-derived CD21 ligand (CD21L), which bind CD21, while antigen in the IC binds on the B cell-BCR. Further, FDC-FcgammaRIIB binds Fc regions of antibodies in IC and reduces coligation of BCR and FcgammaRIIB minimizing an inhibitor of B cell activation. Given that Fc receptors and complement receptors bind immunoglobulins and complement fragments of other species, we reasoned that FDC accessory activity should cross MHC and species barriers. This prediction was tested using memory lymphocytes from OVA-immune mice and TT-immune humans in combination with FDC from murine lymph nodes and human tonsils. Human and murine FDC converted IC into potent immunogens (specific antibody increased from background to thousands of ng / ml). MHC barriers did not restrict this activity and human FDC worked with murine lymphocytes and murine FDC worked with human lymphocytes. Furthermore, stimulation via MHC-dependent allogeneic or zenogeneic mechanisms did not promote antibody production by FDC. Polyclonal responses stimulated by lipopolysaccharide and pokeweed mitogen were also promoted (10 - 100-fold) and anti-CD21 blocked FDC activity. These results substantiate the hypothesis that FDC are necessary for strong recall responses and that FDC-CD21L is critical.     

酵母双杂合系统AD端阴离子交换蛋白C-末端表达质粒的构建

利用PCR方法，从阴离子交换蛋白1（AE1）全长cDNA中扩增出约350bp c末端cDNA片段，测序后将其克隆至pGADT7载体上，用醋酸锂法构建好的pADT7-AE1-c末端转染酵母菌HA109，观察其在选择性培养基上的表达情况。结果表明，获得了530bp AE1c－末端cDNA,pGADT7-AE1-c末端对酵母无毒性，不能激活检测基因，可作为酵母双杂合系统中的靶基因。     

Shear Rate Dependence of Ultrasound Backscattering from Blood Samples Characterized by Different Levels of Erythrocyte Aggregation

The objectives were (1) to determine the effect of the erythrocyte aggregation level (wide range of aggregation) and shear rate (which also affects aggregation) on the ultrasound backscattered power, and (2) to evaluate the reproducibility of the ultrasound method. Experiments were performed under steady flow (100–1250 ml/min) in a 12.7 mm diameter vertical tube. Doppler ultrasound at 10 MHz was used to measure simultaneously the velocity and the backscattered power across the tube. For each radial position, the shear rate was computed from the derivative of the velocity profile. The backscattered power decayed exponentially as a function of the shear rate, and for a given shear rate, the power increased monotonically with the level of aggregation measured by laser reflectometry. Using blood samples simulating hypo-, normal, and hyperaggregating erythrocytes, the power of the ultrasound signal varied respectively by –7.8, –13.2, and –16.1 dB as a function of the shear rate (from 0.4 to 50 s^–1). The reproducibility of the backscattered power was 5.5 dB, which is less than the variations observed as a function of the shear rate. In conclusion, ultrasound backscattering is sensitive to the level of erythrocyte aggregation. At a first glance, ultrasound seems less accurate when compared to laser reflectometry but it is suggested that this is because ultrasound backscattering may be sensitive to structural aggregate changes that are not detected by the laser method. © 2000 Biomedical Engineering Society. PAC00: 8718-h, 8719Tt, 8750Kk     

Injection of substance P (SP) N-terminal fragment SP(1-7) into the ventral tegmental area modulates the levels of nucleus accumbens dopamine and dihydroxyphenylacetic acid in male rats during morphine withdrawal

The biologically active substance P (SP) N-terminal metabolite SP_1–7 has been reported to modulate several neural processes such as learning, locomotor activity and reaction to opioid withdrawal. Although all these processes are believed to be associated with dopaminergic transmission no evidence of an interaction between SP_1–7 and dopamine in the case of morphine withdrawal has so far been reported. Therefore, in this work we applied in vivo microdialysis to investigate the effect of SP_1–7 injection into the ventral tegmental area on dopamine release in nucleus accumbens of male rats during naloxone precipitated morphine withdrawal. The result showed that the heptapeptide enhances dopamine release and also elevates the level of the dopamine metabolite dihydroxyphenylacetic acid in this brain area. It was suggested that the observed action of the SP fragment on the dopamine system represents the underlying mechanism for a previously observed ability of SP_1–7 to counteract the aversion response to morphine withdrawal.     

Defective Treg response in acute kidney injury was caused by a reduction in TIM-3+ Treg cells

Background: Despite years of research, the treatment of acute kidney injury (AKI) remains a significant challenge. Animal studies presented causal links between elevated regulatory T cell (Treg) response and better prognosis in AKI. Previous studies in mice and humans showed that TIM-3+ Treg cells were more potent than TIM-3- Treg cells. In this study, we investigated the role of TIM-3 in Treg in AKI patients.

Methods: Peripheral blood from AKI patients and healthy controls were gathered, and TIM-3+ Treg subset was examined.

Results: Compared to healthy controls, the AKI patients presented a significant upregulation in the frequency of circulating CD4+CD25+ T cells; however, the majority of this increase was from the CD4+CD25+TIM-3- subset, and the frequency of CD4+CD25+TIM-3+ T cells was downregulated in AKI patients. In both healthy controls and AKI patients, the CD4+CD25+TIM-3+ T cells expressed higher levels of Foxp3, and were more potent at expressing LFA-1, LAG-3, CTLA-4, IL-10 and TGF-β. In addition, the CD4+CD25+TIM-3+ T cells from both healthy controls and AKI patients presented higher capacity to suppress CD4+CD25- T cell proliferation than the CD4+CD25+TIM-3- T cells. Interestingly, the total CD4+CD25+ T cells from AKI patients presented significantly lower inhibitory capacity than those from healthy controls, indicating that the low frequency of CD4+CD25+TIM-3+ T cells was restricting the efficacy of the Treg responses in AKI patients.

Conclusions: We demonstrated that TIM-3 downregulation impaired the function of Treg cells in AKI. The therapeutic potential of CD4+CD25+TIM-3+ T cells in AKI should be investigated in future studies.     

Combined use of positive and negative immunomagnetic isolation followed by real-time RT-PCR for detection of the circulating tumor cells in patients with colorectal cancers

To establish a novel molecular diagnostic method of detecting circulating tumor cells (CTCs) LS174T colon cancer cells were serially diluted with normal blood. Additional peripheral blood samples were collected from 25 patients with colorectal carcinoma. Mononuclear cells (MNCs) were collected, equally divided into four parts, and then cancer cells were enriched by four methods: method A, nonimmunobead method; method B, negative immunobead method: CD45 immunomagnetic beads were used to deplete the leukocytes; method C, positive immunobead method: Ber-EP4 immunomagnetic beads were used to enrich cancer cells; method D, negative-and-positive immunobead method: CD45 immunomagnetic beads were first used to deplete the leukocytes from MNC and then Ber-EP4 immunomagnetic beads were used to enrich cancer cells. Finally, real-time quantitative RT-PCR was used to monitor mRNA expression of ₂-mircoglobulin (2M) and carcinoembryonic antigen (CEA). The relative CEA mRNA values were corrected with reference to 2M mRNA, to CEA mRNA/2M mRNA ratios according to a CEA mRNA external standards prepared with tenfold serial dilutions (1–10⁴ IS174T cells) of cDNA and 2M mRNA external standards prepared with tenfold serial dilutions (10²–10⁷ leukocytes) of cDNA. In recovery experiments a significant correlation between the number of cancer cells and CEA mRNA expression was found when CD45 or Ber-EP4 immunomagnetic beads were used alone. A highly significant correlation was found when CD45 and Ber-EP4 immunomagnetic beads were used successively. The sensitivity of method D was one cancer cell per milliliter of blood. Circulating cancer cells were detected in 19 of 25 patients with colorectal cancers. The relative CEA mRNA value obtained by method D was the smallest. The positive detection rate of circulating cancer cells in patients at Dukes B, C, and D stages were 25.0% (1/4), 83.3% (10/12), and 88.9% (8/9). Combinative use of immunomagnetic isolation followed by real-time RT-PCR is a useful technique to detect circulating tumor cells in patients with colorectal carcinomas. Applying negative and positive immunomagnetic beads successively yields the highest correlation with amount of tumor cells.     

The signaling pathways induced by neutrophil-endothelial cell adhesion

Adhesion of neutrophils to vascular endothelial cells (ECs), mediated by the interaction of CD11/CD18 and intercellular adhesion molecule-1 (ICAM-1), is often required for neutrophil transmigration across endothelium during most inflammatory responses. Induction of intracellular signaling in neutrophils as a result of adhesion has been recognized for many years. Recent studies demonstrated that neutrophil-endothelial adhesion also activates ECs. Examples of neutrophil adherence-induced changes in ECs include increases in intracellular Ca(2+), production of reactive oxygen species, and actin cytoskeleton changes. These changes result, in part, from ligation of EC adhesion molecules. This review article focuses on the signaling events that occur in ECs during neutrophil adhesion and the role of EC adhesion molecules, particularly ICAM-1, in the initiation of these signaling events in ECs. The evidence to date describing the molecular basis of ICAM-1-induced signaling will be summarized. Finally, the potential physiological roles of these signaling events induced by EC adhesion molecules in mediating neutrophil migration will be addressed.     

同步录像脑电图在癫（疒/间）外科诊疗中的应用价值

目的：探讨同步录像脑电图（Video-EEG)在癫痫外科诊疗中的意义。方法；对30例难治性癫痫患者进行普通脑电图（REEG）、Video-EEG、术中皮层脑电图（ECoG)及神经影像学检查（CT或（和）MRI），并比较其结果。结果：30例患者中，REEG监测到棘波19例，检出率63％；Video-EEG监测到棘波29例，检出率97％；神经影像学检查发现23例有异常病灶（77％），其中20例定位与Video-EEG完全一致（87％）。在30例手术患者中，29例ECoG的癫痫灶定位与术前Video-EEG一致（97％）。结论：Video-EEG对于癫痫的定位诊断以及指导手术治疗有相当重要的临床价值。     

转录调控蛋白PrfA对两组新近发现的单核细胞增生李斯特菌基因的体外转录作用的研究

目的研究转录调控蛋白PrfA对两组新近发现的单核细胞增生李斯特菌基因的体外转录作用。方法利用本室近年来建立的体外转录系统,对两组基于转录基因组体内研究发现的5个可能的受PrfA不同调节的单核细胞增生李斯特菌基因进行了体外转录活性的研究。结果第一组中的hpt基因的体外转录活性受PrfA正调节,而其它4个基因既不被PrfA正调节也不被负调节。结论除hpt基因外,其它4个基因体外转录结果与体内实验不相一致,说明PrfA在体内可能通过复杂多样的非直接方式、或者还需要一些目前未知的因子来调控这些新近发现的基因的表达。     

Optimization of combined electron and photon beams for breast cancer

Recently, intensity-modulated radiation therapy and modulated electron radiotherapy have gathered a growing interest for the treatment of breast and head and neck tumours. In this work, we carried out a study to combine electron and photon beams to achieve differential dose distributions for multiple target volumes simultaneously. A Monte Carlo based treatment planning system was investigated, which consists of a set of software tools to perform accurate dose calculation, treatment optimization, leaf sequencing and plan analysis. We compared breast treatment plans generated using this home-grown optimization and dose calculation software for different treatment techniques. Five different planning techniques have been developed for this study based on a standard photon beam whole breast treatment and an electron beam tumour bed cone down. Technique 1 includes two 6 MV tangential wedged photon beams followed by an anterior boost electron field. Technique 2 includes two 6 MV tangential intensity-modulated photon beams and the same boost electron field. Technique 3 optimizes two intensity-modulated photon beams based on a boost electron field. Technique 4 optimizes two intensity-modulated photon beams and the weight of the boost electron field. Technique 5 combines two intensity-modulated photon beams with an intensity-modulated electron field. Our results show that technique 2 can reduce hot spots both in the breast and the tumour bed compared to technique 1 (dose inhomogeneity is reduced from 34% to 28% for the target). Techniques 3, 4 and 5 can deliver a more homogeneous dose distribution to the target (with dose inhomogeneities for the target of 22%, 20% and 9%, respectively). In many cases techniques 3, 4 and 5 can reduce the dose to the lung and heart. It is concluded that combined photon and electron beam therapy may be advantageous for treating breast cancer compared to conventional treatment techniques using tangential wedged photon beams followed by a boost electron field.