Acid phosphatase and non-specific alpha-naphthol acetate esterase activities of monocytes in patients with cancer of the digestive tract during surgical treatment

In 22 patients with cancer of the alimentary tract the activities of acid phosphatase and non-specific alpha-naphthol acetate esterase in monocytes were tested. The enzyme activity was tested in the peripheral blood before surgical intervention, in blood from vessels draining the tumour before its excision and in the peripheral blood before surgical intervention, in blood from vessels draining the tumour before its excision and in the peripheral blood 2--3 weeks after tumour excision. In parallel tests the enzyme activity was estimated in the peripheral blood of 22 healthy individuals. The study indicates that the non-specific alpha-naphthol acetate esterase activity of monocytes derived from patients with cancer and control group did not show a marked difference. The acid phosphatase activity in monocytes derived from a tumour efferent vessel was found to be higher in majority of the cases than the activity of this enzyme in monocytes derived from the peripheral blood. After removing the tumour the acid phosphatase activity of monocytes was elevated in half of the cases. It seems possible that the increase of acid phosphatase activity in monocytes derived from cancer patients may be due to the activation of monocytes in contact with cancer antigens or antigen-antibody complexes.     

Progressive myelopathy in older German shepherd dogs. II. Presence of circulating suppressor cells

Progressive myelopathy in the German shepherd dog is a degenerative neurologic disease of unknown etiology. Results presented in a previous study indicated a depression in the response to thymus-dependent mitogens by peripheral blood leukocytes obtained from dogs with progressive myelopathy. Data presented here indicate that this depressed response to mitogens was associated with the presence of peripheral blood suppressor cells. Suppressor cell activity was detected in dogs that were severely affected with PM, but was not apparent in dogs that were mildly affected. Peripheral blood leukocytes obtained from dogs with progressive myelopathy suppressed the mitogenic response to autologous lymph node cells as well as allogeneic normal canine peripheral blood cells. The suppressor cells had the capacity to suppress mixed leukocyte reactions. Suppressor cell activity was radioresistant. Both nylon wool-adherent and -nonadherent peripheral blood leukocyte populations contained suppressor activity. Suppressive activity diminished after incubation of the suppressor cells with indomethacin, suggesting that suppression may be mediated by the release of prostaglandins. Although a role for peripheral blood suppressor cells in the disease process has not yet been established, it is possible that this abnormal regulatory activity reflects an attempt by the host to control an autoimmune event.     

Human colostral cells. II. Response to mitogens

The ability of colostral lymphocytes to respond to pokeweed mitogen, phytohemagglutinin, or Epstein-Barr virus was examined. None of these mitogens induced colostral cells to differentiate into immunoglobulin-containing cells, either in the absence or in the presence of mitomycin C-treated mononuclear cells or T-cell-enriched populations from peripheral blood. Cocultivation of mononuclear cells from the peripheral blood of normal adults with mitomycin C-treated colostral cells resulted in a marked suppression of the generation of immunoglobulin-containing cells in response to pokeweed mitogen. The inhibitory effect was seen in peripheral blood mononuclear cell:colostral cell ratios of 1:1, 5:1, and 10:1. However, colostral cells had little effect on the ability of peripheral blood lymphocytes to proliferate in response to phytohemagglutinin or to allogeneic stimulation.     

Purifying human Y chromosomes by flow cytometry and sorting

A method of producing an enriched sample of human Y chromosomes from peripheral blood lymphocytes is described. Metaphase chromosomes were prepared from peripheral blood lymphocytes donated by 17 normal male individuals. A suspension of chromosomes in a polyamine buffer was produced from each sample, stained with the fluorescent dye Hoechst 33258, and passed through a flow cytometer and sorter. Following analysis of the 17 fluorescence distributions, a single donor was found giving a separate peak corresponding to the Y chromosome. Seventy percent of the chromosomes sorted from this peak were identified as Y chromosomes. Batches of a million Y chromosomes were produced from each of several 40 ml donations of peripheral blood. These were assessed for the amount of Y DNA present and used to construct a DNA library.     

Inhibition of pancreatic somatostatin release in response to glucose

In 5 anaesthetized pigs blood was sampled both from the pancreatic venous effluent and from the peripheral venous circulation. Insulin and somatostatin were determined by radioimmunoassay. The somatostatin level in pancreatic venous blood was increased by a factor 2 as compared to the peripheral blood. Concomitant with the increase in insulin after intravenous glucose a significant decrease in somatostatin immunoreactivity was found, supporting the hypothesis that endogenous somatostatin participates in the local regulation of insulin release.     

CD4 positive Leu-8 negative helper-inducer T cells predominate in the human intestinal lamina propria

The regulatory function of peripheral blood CD4 T cells correlates with the presence or absence of the membrane glycoprotein recognized by anti-Leu-8 antibody; CD4,Leu8- T cells help Ig synthesis and CD4,Leu-8+ T cells suppress Ig synthesis. In contrast to CD4 T cells from the peripheral blood and organized gut-associated lymphoid tissues, intestinal lamina propria CD4 T cells were found to have diminished expression of the Leu-8 Ag. Therefore, studies were performed to determine whether the decreased expression of the Leu-8 Ag on lamina propria CD4 T cells correlates with a difference in the ability of peripheral blood and lamina propria CD4 T cells to regulate PWM-stimulated Ig synthesis. At high T cell to non-T cell ratios, the helper function of lamina propria CD4 T cells was significantly higher than that of peripheral blood CD4 T cells. When CD4 T cells were incubated with anti-Leu-8 antibody, the suppressor function of peripheral blood CD4 T cells was increased, but lamina propria CD4 T cells did not suppress Ig synthesis. No difference was found between the helper function of CD4,Leu-8- T cells and the suppressor function of CD4, Leu-8+ T cells isolated from either the peripheral blood or the lamina propria. Thus, the difference in the regulatory function of CD4 T cells from the peripheral blood and the lamina propria is due to the quantitative difference in CD4,Leu-8+ T cells in these sites. Consequently, the intestinal lamina propria is a site enriched in CD4,Leu-8- T cells which predominantly mediate help for Ig synthesis.     

Leukaemic peripheral blood plasma and bone marrow plasma: comparison of influence on lymphocyte proliferation

Abstract. Peripheral blood plasma from some children with untreated acute lymphoblastic leukaemia (ALL) exerted an inhibitory effect in vitro on phytohaemagglutinininduced lymphocyte transformation of normal peripheral blood lymphocytes. This occurred at concentrations beyond that required for optimal response as judged by reduction of blast cell formation and tritiated thymidine and tritiated uridine incorporation into DNA and RNA, respectively. In contrast, bone marrow plasma from these patients was non-inhibitory or contained significantly less inhibitory activity. Bone marrow plasma from the majority of healthy controls was superior to their peripheral blood plasma in enhancing phytohaemagglutinin-induced mitogenesis. The difference between an individual's bone marrow- and peripheral blood-derived plasma in enhancing proliferation of patient and healthy control cells was significantly greater amongst the patients than the healthy control group; this was attributed mainly to the increased inhibitory activity of ALL peripheral blood plasma compared with normal plasma. Medium conditioned by phytohaemagglutinin-stimulated normal peripheral blood lymphocytes was effective in neutralizing the inhibitory activity of ALL peripheral blood plasma. Taken together, these in vitro results are at least suggestive that in vivo , in healthy subjects, the rapidly proliferating cells in the bone marrow and the 'resting' blood cells in the circulation may be under the influence of a fine balance of different types and/or levels of humoral growth stimulatory and inhibitory factors and that in ALL an unstable balance of these factors exists. The decreased proliferation of circulating blast cells compared with bone marrow blasts in ALL may be attributed, at least in part, to exposure to the different levels of inhibitor(s) in the circulation and bone marrow as demonstrated in vitro by our results.     

人外周血单核细胞的分离和电穿孔法转染

目的:建立分离纯化人外周血单核细胞及转染的有效方法。方法:应用基于HISTOPAQUE的密度梯度离心及抗体标记纯化的方法分离单核细胞,并以电穿孔技术转染绿色荧光蛋白(GFP)表达质粒、肿瘤坏死因子受体相关因子6(TRAF6)小干扰RNA。结果:基于外周血单核细胞的表面标志分子CD14的流式细胞分析表明得到了高纯度(89.95%)的外周血单核细胞;GFP荧光照片显示GFP表达质粒的转染效率为60%～70%;免疫印迹显示TRAF6的表达抑制(90%)下调了脂多糖(LPS)对JNK的激活,说明通过电穿孔技术有效递送了TRAF6小干扰RNA,表明单核细胞若缺少TRAF6将抑制LPS-TLR4信号通路的激活。结论:建立了一种高效的从人外周血中分离原代单核细胞的方法,且应用基于NucleoFectorⅡ的电穿孔技术实现了对此原代单核细胞的高效转染,为进一步外周血单核细胞的相关功能研究奠定了方法学基础。     

Transplantation of hematopoietic stem cells from the peripheral blood

Hematopoietic stem cells can be collected from the peripheral blood. These hematopoietic stem cells (HSC), or better progenitor cells, are mostly expressed as the percentage of cells than react with CD34 antibodies or that form colonies in semi-solid medium (CFU-GM). Under steady-state conditions the number of HSC is much lower in peripheral blood than in bone marrow. Mobilization with chemotherapy and/or growth factors may lead to a concentration of HSC in the peripheral blood that equals or exceeds the concentration in bone marrow. Transplantation of HSC from the peripheral blood results in faster hematologic recovery than HSC from bone marrow. This decreases the risk of infection and the need for blood-product support. For autologous stem-cell transplantation (SCT), the use of peripheral blood cells has completely replaced the use of bone marrow. For allogeneic SCT, on the other hand, the situation is more complex. Since peripheral blood contains more T-lymphocytes than bone marow, the use of HSC from the peripheral blood increases the risk of graft-versus-host disease after allogeneic SCT. For patients with goodrisk leukemia, bone marrow is still preferred, but for patients with high-risk disease, peripheral blood SCT has become the therapy of choice.     

miR-139-5p在前列腺癌患者外周血中的表达及临床意义

目的:研究miR-139-5p在前列腺癌患者外周血中的表达及临床意义。方法:收集2015年4月至2016年9月于我院进行诊治的65例前列腺疾病患者和20例男性健康志愿者外周血样本,使用RT-q PCR方法检测各组miR-139-5相对表达量,统计分析前列腺癌患者外周血miR-139-5p水平与临床特征相关性,使用ROC曲线分析外周血miR-139-5p诊断前列腺癌的临床价值。结果:与良性增生组(n=15)患者和对照组(n=20)健康志愿者相比,前列腺癌组(n=50)患者外周血中miR-139-5p相对表达量均显著升高(P均0.05)。中高分化、转移癌、Gleason评分高危前列腺癌患者外周血miR-139-5p相对表达量显著高于低分化、原位癌和Gleason评分中危的前列腺癌患者(P均0.05)。外周血miR-139-5p在区分前列腺癌和良性前列腺增生或健康人中特异性和敏感性均较高,ROC曲线下面积为0.942(95%CI:0.0.785~0.971)。结论:miR-139-5p在50例前列腺癌患者外周血中呈高表达,或可作为非侵入性前列腺癌诊断标志物。     

Differential synthesis of 5-lipoxygenase in peripheral blood and synovial fluid neutrophils in rheumatoid arthritis

In order to investigate 5-lipoxygenase enzyme regulation in neutrophils during an inflammatory reaction, we studied 5-lipoxygenase mRNA levels, as well as de novo enzyme synthesis, in resting and activated neutrophils isolated from normal individuals and patients with rheumatoid arthritis. The approach used was to analyze these activities in resting peripheral blood neutrophils of normal individuals on the one hand and in peripheral blood and matched synovial fluid neutrophils isolated from patients with rheumatoid arthritis on the other hand. Our first observation was that resting peripheral blood neutrophils of either normal individuals or patients show detectable levels of 5-lipoxygenase mRNA and are able to synthesize the enzyme de novo. Our second observation was that inflammatory activated neutrophils from synovial fluid reveal lower 5-lipoxygenase mRNA levels and enzyme synthesis than do the patient-matched peripheral blood cells. This is in spite of the fact that, for other proteins, synovial fluid neutrophils are equally or more active than their peripheral blood counterparts. We conclude that peripheral blood neutrophils are capable of synthesizing the enzyme, thus ensuring the turnover of the protein. Furthermore, complex regulatory mechanisms appear to take place in response to inflammation as it occurs in synovial fluids of patients with rheumatoid arthritis, leading to decreased mRNA levels and enzyme synthesis. Possible mechanisms of regulation are discussed and are presently under investigation.     

Effects of a cool environment on the health of female office workers and students

OBJECTIVE: The aim of this study was to evaluate the effect of a cool environment on the peripheral skin blood flow and subjective thermal sensations of female office workers and female students. METHODS: The subjects were 26 female bank employees (mean age, 38 years) who worked in a cool environment and 10 female college students (mean age, 22 years). The peripheral skin blood flow was measured using a laser Doppler blood flow meter. In each bank employee, peripheral skin blood flow was measured at three time points during the workday in the medical treatment room at their workplace. In the college students, peripheral skin blood flow was measured every hour between 9:00 and 17:00 in a laboratory. In both the medical treatment room and the laboratory, the room temperature was controlled at 24-26 degrees C with a relative humidity of 55+/-10%. The bank employees and students were each divided into those with hypersensitivity to cold (Group A) and those without hypersensitivity to cold (Group B). RESULTS: When the 10 college students were in the cool environment (24-26 degrees C), their peripheral skin blood flow generally decreased over time. The rate of decrease of this blood flow was greater in Group A than in Group B. In the female bank employees, the peripheral skin blood flow was the lowest at 12:00 (before lunch), was increased at 13:00 (after lunch), and then was decreased at 17:30. However, the degree of the increase from before lunch to after lunch in Group A was about half of that in Group B. CONCLUSION: Among female office workers and students, a cool environment reduced the peripheral skin blood flow of individuals with hypersensitivity to cold to a greater degree than in those without hypersensitivity to cold.     

Biological phenotype of human immunodeficiency virus type 1 clones at different stages of infection: progression of disease is associated with a shift from monocytotropic to T-cell-tropic virus population.

The composition of human immunodeficiency virus type 1 (HIV-1) clonal populations at different stages of infection and in different compartments was analyzed. Biological HIV-1 clones were obtained by primary isolation from patient peripheral blood mononuclear cells under limiting dilution conditions, with either blood donor peripheral blood lymphocytes or monocyte-derived macrophages (MDM) as target cells, and the biological phenotype of the clones was analyzed. In asymptomatic individuals, low frequencies of HIV-1 clones were observed. These clones were non-syncytium inducing and preferentially monocytotropic. In individuals progressing to disease, a 100-fold increase in frequencies of productively HIV-1-infected cells was observed as a result of a selective expansion of nonmonocytotropic clones. In a person progressing to AIDS within 19 months after infection, only syncytium-inducing clones were detected, shifting from MDM-tropic to non-MDM-tropic over time. From his virus donor, a patient with wasting syndrome, only syncytium-inducing clones, mostly non-MDM-tropic, were recovered. Parallel clonal analysis of HIV-1 populations in cells present in bronchoalveolar lavage fluid and peripheral blood from an AIDS patient revealed a qualitatively and quantitatively more monocytotropic virus population in the lung compartment than in peripheral blood at the same time point. These findings indicate that monocytotropic HIV-1 clones, probably generated in the tissues, are responsible for the persistence of HIV-1 infection and that progression of HIV-1 infection is associated with a selective increase of T-cell-tropic, nonmonocytotropic HIV-1 variants in peripheral blood.     

Differential T cell responses to Plasmodium chabaudi chabaudi in peripheral blood and spleens of C57BL/6 mice during infection

The definition of the immune status of a person is often taken as the responses obtained from lymphocytes isolated from peripheral blood. We therefore analyzed in a mouse model of Plasmodium chabaudi chabaudi the response of T lymphocytes taken from peripheral blood and compared it with the spleen during and after a primary erythrocytic infection. Using limiting dilution conditions, no malaria-specific T cell responses could be measured in the peripheral blood for up to 21 days after infection with P. chabaudi, whereas T cells responding to malaria Ag were readily detected in the spleen. This was true for T cells providing help and for those producing IFN-gamma. After clearance of the parasitemias to subpatent levels (75 days), qualitatively similar T cell responses were found in both compartments of the immune system, i.e., the Th cell response predominated over the inflammatory response. These data suggest that during an active infection with Plasmodium, T cell responses in peripheral blood are not necessarily indicators of the immune status.     

Small-Sample Blood Culture Method for Identification of Bacteria in Central Arterial and Peripheral Blood

A blood culture technique that utilized small arterial blood samples or peripheral capillary blood was tested in beagle dogs and pig-tailed macaque monkeys. A bolus of 2.0 x 10(7)Escherichia coli (ATCC 25922) was injected intravenously into five animals of each species. Blood samples were taken before injection of the organisms and 10, 15, 20, 30, 60, and 120 min after injection. Arterial blood samples (2.0 and 0.2 ml) and peripheral capillary samples (0.14 ml) were taken at each sampling time. Pour plates were prepared from arterial blood for colony counts. All three blood sampling methods were equally effective in detecting sepsis when 10 or more organisms per ml of blood were present. Below this level, the 2.0-ml sample was more effective. Contamination of the peripheral sample with air or skin contaminants was a problem.     

外周全血中与老化相关的DNA甲基化标记的来源

机体老化与癌症、神经退行性疾病等许多复杂疾病相关。目前,研究者已在外周全血中识别了大量的与老化相关的DNA甲基化标记,这些标记可能反映外周血白细胞在机体老化过程中发生的变化,也可能反映外周血中与年龄相关的细胞构成比例的变化。文章利用3组正常个体外周全血DNA甲基化谱,采用Spearman秩相关分析识别了与老化相关的CpG甲基化位点(age-related DNA methylation CpG sites, arCpGs)并评价了其可重复性;利用去卷积算法估计了各外周血样本中髓性和淋巴性细胞的比例并分析了其与年龄的相关性;比较了在外周全血、CD4+T细胞和CD14+单核细胞中识别的arCpGs的一致性。结果显示,在独立外周全血数据中识别的arCpGs具有显著的可重复性(超几何检验,P=1.65×10^-11)。外周血髓性和淋巴性细胞的比例分别与年龄显著正、负相关(Spearman秩相关检验,P<0.05,r≤0.22),它们间DNA甲基化水平差异较大的CpG位点倾向于在外周全血中被识别为arCpGs。在CD4+T细胞中识别的arCpGs与在外周全血中识别的arCpGs显著交叠(超几何检验,P=6.14×10^-12),且99.1%的交叠位点在CD4+T细胞及外周全血中的DNA甲基化水平与年龄的正、负相关性一致。尽管在CD14+单核细胞中识别的arCpGs与在外周全血中识别的arCpGs并不显著交叠,但是在交叠的51个arCpGs中,有90.1%的位点在CD14+单核细胞、外周全血以及CD4+T细胞中的DNA甲基化水平与年龄的正、负相关性一致,提示它们可能主要反映细胞间共同的改变。在外周全血中识别的arCpGs主要反映某些白细胞共同或特异的DNA甲基化改变,但是也有一部分反映外周血细胞比例构成的变化。     

Myelin basic protein-stimulated rosette-forming T cells in multiple sclerosis

A subpopulation of T lymphocytes sensitized to human myelin basic protein in peripheral blood of patients with multiple sclerosis, central nervous system (CNS) tumors, and cerebrovascular accidents was demonstrated by the antigen-stimulated, rosette-forming T cell assay. A significant increase in the percent of active rosette-forming T cells was detected after in vitro exposure of peripheral blood lymphocytes to human myelin basic protein but not to histones. In contrast, peripheral blood lymphocytes from healthy controls and from patients with benign and malignant breast diseases were unresponsive to stimulation by either antigen. These results demonstrate a functionally active T-lymphocyte subpopulation sensitized to myelin basic protein in patients with multiple sclerosis and in patients with certain other CNS diseases.     

Action of adrenalin on the circulation of the murine Plasmodium developing stages, in different blood compartments

Adrenalin was used to investigate in vivo the circulation of the different stages of rodent Plasmodium present in the blood. A single dose of adrenalin injected to mice infected with P. yoelii resulted immediately in i) a diminution of the parasitaemia of approximately 50% in the peripheral large vessels (estimated in tail blood films), as well as in the capillaries (estimated in smears of blood collected from a fed Anopheles), and ii) an increased parasitaemia in blood collected by cardiac puncture from the right heart. The numbers of young stages of P. yoelii in the peripheral blood were initially somewhat reduced but, unexpectedly, midterm trophozoites were preferentially expelled from the peripheral blood into major organs like the heart. With P. vinckei, parasitaemia decreased only when midterm trophozoites predominated, and with P. chabaudi no effect was observed at any time. We propose that midterm trophozoites, by their increased surface area, as compared to rings, and their flexibility which contrasts with the rigid schizonts, are particularly susceptible to haemodynamic perturbations.     

Lipoxygenase mRNA in rabbit reticulocytes. Its isolation, characterization and translational repression

The synthesis of the erythroid lipoxygenase, an enzyme which is of importance for the degradation of mitochondria during the maturation of reticulocytes to erythrocytes, was studied in reticulocytes from bone marrow and in density-separated fractions from peripheral blood of anemic rabbits. Lipoxygenase mRNA was enriched to about 75% by digestion of polysomes with protease K, poly(U)-Sepharose chromatography and repeated sucrose gradient centrifugation. From sucrose gradient centrifugation, electrophoresis and electron microscopy a molecular weight of about 10(6) was calculated. Synthesis of lipoxygenase is absent in erythroblasts, in very young reticulocytes obtained from bone marrow, or in the lightest fractions of reticulocytes from the peripheral blood. More mature blood reticulocytes show a considerable synthesis of the enzyme. The induction of the synthesis of the lipoxygenase seems to be initiated when reticulocytes have reached the peripheral blood. It is shown that lipoxygenase mRNA is present in reticulocytes as a translationally inactive free cytoplasmic messenger ribonucleoprotein (mRNP) particle. After deproteinization isolated mRNA obtained from masked mRNP codes for authentic lipoxygenase in a cell-free protein-synthesizing system of reticulocytes.     

The persistence of micronucleated erythrocytes in the peripheral circulation of normal and splenectomized Fischer 344 rats: implications for cytogenetic screening

Micronucleated erythrocytes are selectively removed from the peripheral circulation of normal rats. Splenectomy prevents this selective removal. In normal rats treated daily for 20 days with 0.2 mg/kg triethylenemelamine (TEM), micronucleated normochromatic (mature) erythrocytes did not accumulate in peripheral blood. In these same animals, the frequencies of micronucleated cells among polychromatic (newly formed) erythrocytes increased from 0.21 to 5.25 per thousand in peripheral blood and from 1.75 to 31.5 per thousand in bone marrow. Since both control and induced frequencies in peripheral blood were approximately 15% of those in bone marrow, the removal appears to be equally efficient for cells containing either spontaneously occurring or clastogen-induced micronuclei. In splenectomized rats treated daily for 11 days with 0.2 mg/kg TEM, the frequency of micronucleated normochromatic erythrocytes (NCEs) in the peripheral blood rose rapidly to 9 times the control value and remained elevated for 50-55 days, indicating a life span approximately equivalent to that of normal erythrocytes. Among splenectomized rats exposed to either 0.15 mg/kg triethylenemelamine, 6.5 mg/kg cyclophosphamide, or 300 mg/kg urethane for periods exceeding the erythrocyte life span, the incidences of micronucleated NCEs in the peripheral blood rose steadily from a control value of 1.0 per thousand to maximum values of 15.0, 12.7 and 8.9 per thousand, respectively. During these extended exposures, the mean frequencies of micronucleated polychromatic erythrocytes (PCEs) in peripheral blood increased from a spontaneous value of 0.9 per thousand to 23.0, 13.0 and 6.6 per thousand, respectively, reflecting the frequencies among PCEs in the bone marrow and approximating the maximum values among NCEs in the peripheral blood. Thus, the frequency of micronucleated erythrocytes in the peripheral blood of splenectomized rats can be used as an index of both acute and cumulative chromosomal damage, while in normal rats the use of peripheral blood for cytogenetic monitoring is restricted by the selective removal of these micronucleated cells.