Pharmacologic action of isoxsuprine in cutaneous and myocutaneous flaps

The therapeutic effects of isoxsuprine on skin capillary blood flow and viability were studied in arterial buttock flaps, latissimus dorsi myocutaneous flaps, and random skin flaps in pigs. It was observed that parenteral isoxsuprine increased capillary blood flow to the skin of arterial buttock flaps and the skin and muscle of latissimus dorsi myocutaneous flaps in a dose-response manner, with a maximum vascular effect observed at 1.0 mg/kg. However, this maximum effective dose of isoxsuprine did not have any significant effect on skin viability in the cutaneous and myocutaneous flaps compared with the control. Examination of the distribution of capillary blood flow within the flaps at varying distances from the pedicle revealed that isoxsuprine did not increase capillary blood flow or perfusion distance in the distal portion of the skin of arterial buttock flaps, latissimus dorsi myocutaneous flaps, and random skin flaps. The increased capillary blood flow as a result of isoxsuprine treatment was limited only to the arterial portion of the arterial buttock flaps and latissimus dorsi flaps. Therefore, it is concluded that isoxsuprine alone is not effective in augmentation of skin viability in cutaneous and myocutaneous flaps. The pharmacologic action of isoxsuprine on the vasculature in the skin and muscle of flaps was also discussed.     

The differing effects of isoxsuprine on muscle flap and skin flap survival in the pig

The pretreatment of pigs with isoxsuprine for two weeks resulted in a significant increase in the surviving length of sartorius muscle flaps when compared with untreated flaps on the same animal. However, there was no improvement in the surviving length of neurovascular island skin flaps when these animals were treated with isoxsuprine. The difference in these results is thought to be due to the action of isoxsuprine on the blood supply of skeletal muscle.     

Drug treatment and flap survival

Some investigators found that isoxsuprine, propranolol, or heparin would increase skin-flap survival in loose-skinned animals. We evaluated the effects of these three drugs in the pig, an animal with skin circulation similar to that of humans. Four hundred ventrally based skin flaps that have a proximal axial portion and a distal random portion were made on the flanks of 40 pigs. There were eight study groups: control, isoxsuprine preoperatively and postoperatively, propranolol preoperatively and postoperatively, isoxsuprine postoperatively only, propranolol postoperatively only, heparin, single-stage surgical delay, and two-stage surgical delay. Flap survival was improved by the two-stage surgical delay when compared with the control flaps, flaps from pigs receiving a drug, or flaps from pigs having a single-stage surgical delay (p less than 0.001). When compared with the control flaps, neither isoxsuprine, propranolol, heparin, nor single-stage surgical delay significantly increased flap survival.     

Identification of Isoxsuprine Hydrochloride as a Neuroprotectant in Ischemic Stroke through Cell-Based High-Throughput Screening

Stroke is a leading cause of death and disability and treatment options are limited. A promising approach to accelerate the development of new therapeutics is the use of high-throughput screening of chemical libraries. Using a cell-based high-throughput oxygen-glucose deprivation (OGD) model, we evaluated 1,200 small molecules for repurposed application in stroke therapy. Isoxsuprine hydrochloride was identified as a potent neuroprotective compound in primary neurons exposed to OGD. Isoxsuprine, a β₂-adrenergic agonist and NR2B subtype-selective N-methyl-D-aspartate (NMDA) receptor antagonist, demonstrated no loss of efficacy when administered up to an hour after reoxygenation in an in vitro stroke model. In an animal model of transient focal ischemia, isoxsuprine significantly reduced infarct volume compared to vehicle (137±18 mm³ versus 279±25 mm³, p<0.001). Isoxsuprine, a peripheral vasodilator, was FDA approved for the treatment of cerebrovascular insufficiency and peripheral vascular disease. Our demonstration of the significant and novel neuroprotective action of isoxsuprine hydrochloride in an in vivo stroke model and its history of human use suggest that isoxsuprine may be an ideal candidate for further investigation as a potential stroke therapeutic.     

CXCR7 protein is not expressed on human or mouse leukocytes

Since the discovery that CXCR7 binds to CXCL12/SDF-1α, the role of CXCR7 in CXCL12-mediated biological processes has been under intensive scrutiny. However, there is no consensus in the literature on the expression of CXCR7 protein by peripheral blood cells. In this study we analyzed human and mouse leukocytes and erythrocytes for CXCR7 protein expression, using a competitive CXCL12 binding assay as well as by flow cytometry and immunohistochemistry using multiple CXCR7 Abs. CXCR7(-/-) mice were used as negative controls. Together, these methods indicate that CXCR7 protein is not expressed by human peripheral blood T cells, B cells, NK cells, or monocytes, or by mouse peripheral blood leukocytes. CXCR7 protein is, however, expressed on mouse primitive erythroid cells, which supply oxygen to the embryo during early stages of development. These studies therefore suggest that, whereas CXCR7 protein is expressed by primitive RBCs during murine embryonic development, in adult mammals CXCR7 protein is not expressed by normal peripheral blood cells.     

Interleukin-12 augments a Th1-type immune response manifested as lymphocyte proliferation and interferon gamma production in Leishmania infantum-infected dogs

The dog is the major reservoir for human visceral leishmaniasis caused by Leishmania infantum. Interleukin-12 is considered to have an essential role in the development of both innate and adaptive immunity to Leishmania spp. and other intracellular pathogens. This study focused on the influence of IL-12 in experimental and natural canine visceral leishmaniasis. Responses of peripheral blood mononuclear cells to IL-12, interleukin-10 and Leishmania soluble antigen were evaluated in L. infantum experimentally infected oligosymptomatic beagles, uninfected beagles, naturally infected polysymptomatic dogs, and their matched uninfected controls. Leishmania soluble antigen induced strong peripheral blood mononuclear cells proliferation both in experimentally infected dogs (median stimulation index [SI]=15.01), and in naturally infected dogs (SI=8.86), but not by cells from the control groups. IL-12 addition further enhanced cell proliferation in naturally (SI=14.95), but not in experimentally infected animals. Peripheral blood mononuclear cells from experimentally infected dogs were able to produce significant amounts of IFN-gamma (3.39 ng/ml) upon LSA stimulation, but no such production was detected in cells from naturally infected or control animals. Interestingly, addition of IL-12 reversed the inhibitory effect of LSA on IFN-gamma production by cells from polysymptomatic naturally infected dogs and the uninfected beagles (4.84 and 7.45 ng/ml, respectively), and further increased IFN-gamma production by peripheral blood mononuclear cells from experimentally infected oligosymptomatic dogs (29.28 ng/ml). IFN-gamma mRNA expression correlated well with IFN-gamma production. Addition of IL-10 to Leishmania soluble antigen stimulated peripheral blood mononuclear cells inhibited proliferation and IFN-gamma production in experimentally infected dogs. Thus, the ability of IL-12 to augment IFN-gamma production by peripheral blood mononuclear cells from dogs with experimental or natural symptomatic canine visceral leishmaniasis makes it a good candidate for cytokine therapy in dogs that are refractory to current therapy.     

Effect of xylazine,isoxsuprine, and lidocaine on Doppler sonographic uterine and umbilical blood flow measurements in cows during the last month of pregnancy

The maternal portion of the bovine placenta receives blood mainly from the uterine arteries (AUT) and the fetal portion from the umbilical arteries (AUM). Placental perfusion is crucial for fetal development and undergoes adaptive changes during pregnancy according to the fetal requirements. One goal of this study was to investigate changes in Doppler sonographic measurements of blood vessels that supply blood to the placenta in cows during the last 4 weeks of pregnancy. Another goal was to examine how these measurements are affected by three drugs commonly used in cows at the time of parturition. Nine cows underwent Doppler sonographic examination of the AUT ipsilateral and contralateral to the pregnant horn and one AUM three times per week during the last 4 weeks of gestation. This was followed by the randomized administration of one of the three following experimental drugs per day: isoxsuprine (200 mg/cow, iv), xylazine (2 mg/100 kg, iv), and lidocaine for epidural anesthesia (100 mg/cow). Doppler sonographic examination was repeated 30 minutes after medication. Maternal pulse rate increased during the study period (P < 0.001), and the diameter of the contralateral AUT was smaller in the last week before birth than in the two preceding weeks. The resistance index (RI) of the ipsilateral AUT was smaller in the last week than in the first 2 weeks of the study period. Uterine blood flow volume increased after isoxsuprine by 5% and after epidural anesthesia by 6% (both P ≤ 0.05) and decreased after xylazine by 10% (P < 0.001). Isoxsuprine was the only drug that elevated the blood flow volume in the AUM (P ≤ 0.05). Xylazine increased the RI of both AUT (both P < 0.001) and significantly reduced maternal and fetal pulse frequencies, whereas isoxsuprine significantly reduced the RI of both AUT and the AUM and increased maternal and fetal pulse frequencies. The results reported that Doppler sonographic measurements of uterine and AUM change little in the last month of pregnancy in the cow. Isoxsuprine and epidural anesthesia with lidocaine have the potential to improve uterine perfusion.     

Collection and phagocytic evaluation of uterine neutrophilic leukocytes

The study of phagocytosis in the bovine uterus requires the collection of large numbers of uterine leukocytes. A method of stimulating polymorphonuclear neutrophil (PMN) invasion into the bovine uterus with oyster glycogen was tested. Following glycogen treatment, uterine aspirates contained large numbers (10(6)-10(9)) of viable uterine PMN. In Experiment 1, six cows were treated at one-week intervals by intrauterine infusion with 0.1, 1.0 and 10.0% oyster glycogen, respectively. Uterine lumens were lavaged at 0, 12, 24, 48, 72 and 168 hours post-infusion with each concentration of glycogen. The mean peak response for PMN was at 12 hours post-infusion of glycogen. In Experiment 2, uterine and peripheral blood PMN were collected from each of 12 cows. Phagocytosis was measured as the percent of (32)Phosphorus-labeled Staphylococcus aureus ingested by PMN during a one-hour incubation. Mean (+/- standard deviation) percent phagocytosis was not statistically different between blood (53.6 +/- 11.0%) and uterine (43.8 +/- 13.0%) PMN. This indicates that phagocytic activity was similar for PMN collected either from the uterine lumen or peripheral blood.     

Cytokine profiles in peripheral, placental and cord blood in pregnant women from an area endemic for Plasmodium falciparum

During gestation, inflammatory cytokines are sometimes more abundant than growth-promoting cytokines, and via direct or indirect effects, proinflammatory cytokines lead to intrauterine growth retardation. We used an enzyme-linked immunosorbent assay to measure the concentrations of three proinflammatory cytokines, tumor necrosis factor alpha (TNF-alpha), interleukin-12 (IL-12p40), as well as interleukin-15 (IL-15) and monocyte chemotactic protein-1 (MCP-1), in plasma from peripheral, placental and cord blood of thirty pregnant Gabonese women. All of these women lived in Libreville and Lambaréné, two malaria hyperendemic areas. IL-12p40 concentrations were higher in cord blood than in placental or peripheral blood. The MCP-1 concentration was higher in placental blood, than in peripheral or cord blood. IL-15 concentrations were similar at the three sites. MCP-1 concentrations were higher in the placentas of primiparous women than in those of multiparous women. The highest concentrations were found in infected placentas. IL-15 concentrations were significantly higher in peripheral and placental plasma from uninfected women than in plasma from infected women. Strong positive correlations were found between placental and cord IL-12p40 and IL-15 plasma concentrations. Likewise, a strong positive correlation was found between IL-12p40 and MCP-1 concentrations in cord and peripheral plasma. These results suggest that placental, maternal peripheral and cord blood present different cytokine profiles in response to P. falciparum.     

Regulation of Staphylococcus aureus-mediated activation of interleukin-18 in peripheral blood mononuclear cells.

Bacteria and bacterial antigens strongly induce cytokine secretion by peripheral blood leukocytes and thereby initiate an inflammatory cascade with potentially deleterious consequences for the host. The present study focussed on receptors and signal transduction pathways involved in activation of interleukin (IL)-18 by heat-inactivated Gram-positive Staphylococcus aureus Cowan strain I (SAC). Similarly to IL-12/IL-12p40, IL-10 and IFN-gamma, SAC dose-dependently activated IL-18. Secretion of IL-18 was independent of functional activity of IL-10, IL-12 or IFN-gamma. Lipoteichoic acid (LTA), a structural component of SAC, was not sufficient for activation of IL-18, while it dose-dependently induced IL-10. In contrast to IL-12, blockade of CD14 only partially diminished secretion of IL-18 and did not affect secretion of IL-10, suggesting involvement of other receptors (e.g., Toll-like receptors) in SAC responses. Further down-stream however, secretion of IL-10, IL-12 and IL-18 was uniformly inhibited by blockade of G-protein-mediated kinase activation by mastoparan. Secretion of IL-18 required phosphatidylinositol-3'-kinase, and secretion of IL-12 phosphotyrosine kinase activity. The data demonstrate that SAC potently activates secretion of IL-18 by peripheral blood mononuclear cells with differential involvement of cell-surface receptors and signal transduction pathways as compared to other natural killer- and T cell-promoting cytokines.     

M2b monocytes predominated in peripheral blood of severely burned patients

Severely burned patients were shown to be carriers of M2 monocytes, and all of the monocytes isolated from peripheral blood of severely burned patients (19 of 19 patients) were demonstrated as M2b monocytes (IL-12(-)IL-10(+)CCL1(+) monocytes). Low levels of M2a (IL-12(-)IL-10(+)CCL17(+) monocytes) and M2c monocytes (IL-12(-)IL-10(+)CXCL13(+) monocytes) were demonstrated in peripheral blood of severely burned patients (M2a, 2 of 19 patients; M2c, 5 of 19 patients). M2b, M2a, and M2c monocytes were not detected in peripheral blood of healthy donors. However, M2b monocytes appeared when healthy donor monocytes were cultured in media supplemented with burn patient serum (15%). CCL2 was detected in sera of all burn patients, and M2b monocytes were not generated from healthy donor monocytes cultured with media containing 15% burn patient sera that were previously treated with anti-CCL2 mAb. In addition, M2b monocytes were generated from healthy donor monocytes in cultures supplemented with rCCL2. These results indicate that M2b monocytes are predominant in peripheral blood of severely burned patients who are carriers of CCL2 that functions to stimulate monocyte conversion from resident monocytes to M2b monocytes.     

Study of chemotactic activity developed by neutrophils from rheumatoid arthritis patients

Neutrophils are the predominant cells accumulated in the synovial fluid (SF) of rheumatoid arthritis (RA) patients. Accumulation of neutrophils may be regarded as a possible way by which neutrophils exert cytotoxic functions. The aim of the present study was to analyze the chemotactic response of neutrophils (PMNs) isolated from the peripheral blood or SF of patients with RA by performing the chemotaxis assay, in which N-formyl-methionyl-leucyl-phenylalanine (FMLP) was used as chemotactic agent. Our results showed that FMLP induced response of peripheral blood neutrophils from 12 patients with RA was similar with the response of 15 healthy controls. A decreased chemotactic response to FMLP was, however, observed in PMNs isolated from the SF of RA patients as comlipared with peripheral blood cells. Therefore, this defective chemotactic ability of neutrophil, was inversely correlated with the number of infiltrating cells in SF. These results indicate that chemotactic ability of neutrophils may be reduced after migration to the SF. Because PMNs chemotaxis in vivo has likely occurred in the presence of serum or SF, we tried to simulate the same conditions in vitro. Therefore, we analyzed the effect of serum or SF on the RA-PMNs chemotaxis. Heat-inactivated serum produced a marked reduction of chemotactic activity developed by PMNs isolated from patients with RA. Notably, a significant increase of chemotactic activity was observed when FMLP and serum stimuli were used together, as compared with the same stimuli used alone. The results suggested that complement activation might interfere with neutrophils chemotaxis. SF amplifies the chemotactic activity of PMNs isolated from peripheral blood of RA patients, but does not affect the chemotaxis developed by PMNs isolated from SF. The data might suggest that several components of SF (IL-8, leukotrien B4, thrombin, platelet-activating factor, etc.) could serve as a potent stimulus for recruitment of neutrophils from periphery into the RA joint. In conclusion, serum or SF components seem to contribute to chemotaxis of neutrophils and play a role in differential killing of PMNs and incidence of infection.     

22q11.2 deletion mosaicism in patients with conotruncal heart defects

BACKGROUND: Some patients with conotruncal heart defects (CTDs) have a chromosome 22q11.2 deletion, but we do not know whether patients with CTDs who are missing the peripheral blood-cell chromosome 22q11.2 deletion are also missing the 22q11.2 deletion in myocardial cells, and whether patients with the 22q11.2 deletion can show a different 22q11.2 deletion in peripheral blood cells and myocardial cells due to a postzygotic mutation during the embryonic period. METHODS: A total of 32 Chinese pediatric nonsyndromic CTD patients (21 with tetralogy of fallot [TOF], 9 with double outlet right ventricle [DORV], 1 with pulmonary artery atresia with ventricular septal defect [PAA/VSD], and 1 with congenitally corrected transposition of the great arteries [CCTGA]), 12 females and 20 males ranging in age from 5 months to 7 years, were included in our study. We used fluorescence in situ hybridization (FISH) to find the chromosome 22q11.2 deletion in peripheral blood cells and compared genotypes of 15 short tandem repeat (STR) markers within 22q11.2 between peripheral blood cells and myocardial cells to search for genetic mosaicism of the chromosome 22q11.2 deletion. RESULTS: Three patients, 2 with TOF and 1 with DORV, were determined to have the peripheral blood cell chromosome 22q11.2 deletion. There was no STR genotypic difference observed between peripheral blood cells and myocardial cells in patients with or without the chromosome 22q11.2 deletion. CONCLUSIONS: Genetic mosaicism may not play a major role in the etiology of isolated CTDs.     

Changes in the K562 cell sensitivity to nonspecific lysis by human and rat leukocytes under the influence of sodium butyrate, dimethyl sulfoxide and phorbol-12-myristate-13-acetate

We studied the ability of inducers and inhibitors of erythroid differentiation of K562 leukemia cells, such as sodium butyrate, dimethyl sulfoxide, and phorbol-12-myristate-13-acetate, respectively, to modulate sensitivity of these cells to non-specific lysis (non-restricted with respect to antigens of the major histocompatibility complex) mediated by natural human or rat killer cells. Unfractionated leukocytes from human peripheral blood or rat splenocytes were used as sources of natural killers. The induction of erythroid differentiation by sodium butyrate was accompanied by a significant increase in cell sensitivity to lysis with human peripheral blood lymphocytes; incubation of K562 cells in the mixture of sodium butyrate and dimethyl sulfoxide did not change cell sensitivity to lysis by both types of effector cells. The inhibition of sodium butyrate-induced erythroid differentiation with high doses of phorbol-12-myristate-13-acetate (100 nM; incubation was in the presence of both these agents simultaneously) resulted in an increased cell sensitivity to lysis with rat splenocytes. Incubation of K562 cells in a mixture of sodium butyrate, dimethyl sulfoxide, and phorbol-12-myristate-13-acetate (100 nM) produced greater lysis by human leukocytes, as compared with incubation in the mixture of sodium butyrate and dimethyl sulfoxide.     

Exposure to sodium butyrate, dimethyl sulfoxide, and phorbol-12-myristine-13-acetate alters sensitivity of K562 cells to nonspecific lysis by human or rat leukocytes

We studied the ability of inducers and inhibitors of erythroid differentiation of K562 leukemia cells, such as sodium butyrate, dimethyl sulfoxide, and phorbol-12-myristate-13-acetate, respectively, to modulate sensitivity of these cells to nonspecific lysis (nonrestricted with respect to antigens of the major histocompatibilty complex) mediated by natural human or rat killer cells. Unfractionated leukocytes from human peripheral blood or rat splenocytes were used as sources of natural killers. The induction of erythroid differentiation by sodium butyrate was accompanied by a significant increase in cell sensitivity to lysis with human peripheral blood lymphocytes; incubation of K562 cells in the mixture of sodium butyrate and dimethyl sulfoxide did not change cell sensitivity to lysis by both types of effector cells. The inhibition of sodium butyrate-induced erythroid differentiation with high doses of phorbol-12-myristate-13-acetate (100 nM; incubation was in the presence of both these agents simultaneously) resulted in an increased cell sensitivity to lysis with rat splenocytes. Incubation of K562 cells in a mixture of sodium butyrate, dimethyl sulfoxide, and phorbol-12-myristate-13-acetate (100 nM) produced greater lysis by human leukocytes, as compared with incubation in the mixture of sodium butyrate and dimethyl sulfoxide.     

Identification and Characterization of K12 (SECTM1), a Novel Human Gene That Encodes a Golgi-Associated Protein with Transmembrane and Secreted Isoforms

The investigation of a DNase-hypersensitive site upstream of the CD7 gene on chromosome 17q25 has led to the discovery of a novel human gene designated K12 (SECTM1, the HGMW assignment). This gene spans 14 kb and encodes a 1.8-kb mRNA detected at the highest levels in peripheral blood leukocytes and breast cancer cell lines. The open reading frame predicts a 248-amino-acid protein with the hydropathic characteristics of a type 1a membrane protein. Western blots show that the K12 protein exists as a cluster of bands around 27 kDa, and extractions using nonionic detergents or high pH conditions demonstrate that it behaves as an integral membrane protein. Immunofluorescence localization studies reveal that K12 is not detectable on the cell surface, but instead is found in a perinuclear Golgi-like pattern and colocalizes with a well-known Golgi marker. In addition, an 20-kDa soluble form of the K12 protein derived from the N-terminal domain is specifically secreted by cells into the culture medium. Immunohistochemical analysis of peripheral blood cells shows that K12 is found in leukocytes of the myeloid lineage, with the strongest staining observed in granulocytes and no detectable expression in lymphocytes. Based on its range of expression, its broad structural characteristics that resemble cytokines and growth factors, and the chromosomal location of the gene in an area already associated with myelogenous leukemias and other malignant neoplasms, this study concludes that K12 is a novel molecule with potential importance in hematopoietic and/or immune system processes.     

The micronucleus assay with peripheral blood reticulocytes by acridine orange supravital staining with 1-beta-D-arabinofuranosylcytosine.

Dose-dependent induction of micronuclei with 1-beta-D-arabinofuranosylcytosine (ara-C) was clearly shown in CD-1 mouse peripheral blood reticulocytes (RETs) using an acridine orange (AO) supravital staining method, as well as in the conventional bone marrow assay. The maximum frequencies of micronucleated RETs (MNRETs) in peripheral blood and of micronucleated polychromatic erythrocytes (MNPCEs) in bone marrow were comparable, as shown in two laboratories independently. The maximum frequencies of MNRETs in peripheral blood lagged about 24 and 12 h behind those of MNPCEs in bone marrow in experiments with 24- and 12-h sampling intervals, respectively. The proportion of each type of RET was examined periodically after treatment with ara-C at doses ranging from 6.25 to 50.0 mg/kg. The proportion of type I RETs among total RETs decreased 24 or 48 h after treatment according to the dose level. This suggest that this ratio could be a good indicator of the bone marrow cell toxicity of test chemicals.     

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。