The Effect of Human Cord Blood on SJL/J Mice after Chemoablation and Irradiation and its Possible Clinical Significance

There is evidence from the existing published literature that human umbilical cord blood, when used for purposes of bone marrow transplantation, does not necessarily have to be HLA matched in order to be efficacious. These reports include experimental observations on the ability of human umbilical cord blood to rescue lethally irradiated mice and clinical observations from China wherein HLA mismatched umbilical cord blood has been engrafted successfully in children with malignant disease.

The study reported herein describes an experimental immunocompetent murine model to determine if human umbilical cord blood can be used to improve survival after chemoablation and irradiation. The animals received chemoablation followed by irradiation, and irradiation alone. The presence of human DNA in these mice following injection of human umbilical cord blood cells was determined, and the immunological status of the animals was evaluated. Animals receiving human umbilical cord blood cells after chemoablation and irradiation had a better mean survival at day 50 than animals receiving syngeneic marrow. Human DNA could be found in various organs, particularly the lung, spleen and liver of the mice for the first 30 days. Thereafter, human DNA became more difficult to detect but trace amounts of human DNA could be found up to one year later. The results of mixed lymphocyte reactions and phenotype analyses for murine T cell markers performed after injection of HUCB cells both indicated endogenous repopulation, and relatively intact immune systems in these mice.

Since human umbilical cord blood allowed mice to survive the lethal effects of chemoablation plus irradiation, or irradiation alone, with reconstitution of the animals' own, relatively intact, immune systems, it would appear that HLA mismatched human umbilical cord blood could potentially be used as an adjuvant treatment for patients with advanced malignancies or other diseases for which hematopoietic reconstitution is indicated.     

紫外线照射充氧自血回输对家兔脊髓损伤后血液循环的影响

家兔用改良Allen 氏系统造成不完全性脊髓损伤,随机分为对照组、损伤组和治疗组。分别测定血液和脊髓组织TXB 2、 6-keto-PGF 1 a 和ET-1 含量;以及脊髓组织灰质、白质和总血流量,研究紫外线照射和充氧自血回输疗法（UBIO）治疗脊髓损伤的作用机制。实验发现：损伤组血液和脊髓组织TXB 2 含量升高、6-keto-PGF 1 a 含量下降、ET-1 含量升高。脊髓组织灰质和白质血流量以及脊髓组织总血流量均下降。治疗组用UBIO 治疗后,血液和脊髓组织TXB 2 含量下降、6-keto-PGF 1 a 含量升高、ET-1 含量下降。脊髓组织灰质和白质血流量以及脊髓组织总血流量均上升。实验说明：UBIO 可以改善全身和脊髓组织的血液循环。     

Effect of Human Parathyroid Hormone on Hematopoietic Progenitor Cells in NOD/SCID Mice Co-Transplanted with Human Cord Blood Mononuclear Cells and Mesenchymal Stem Cells

Purpose

We evaluated the effect of human parathyroid hormone (hPTH) on the engraftment and/or in vivo expansion of hematopoietic stem cells in an umbilical cord blood (UCB)-xenotransplantation model. In addition, we assessed its effect on the expression of cell adhesion molecules.

Materials and Methods

Female NOD/SCID mice received sublethal total body irradiation with a single dose of 250 cGy. Eighteen to 24 hours after irradiation, 1×10⁷ human UCB-derived mononuclear cells (MNCs) and 5×10⁶ human UCB-derived mesenchymal stem cells (MSCs) were infused via the tail vein. Mice were randomly divided into three groups: Group 1 mice received MNCs only, Group 2 received MNCs only and were then treated with hPTH, Group 3 mice received MNCs and MSCs, and were treated with hPTH.

Results

Engraftment was achieved in all the mice. Bone marrow cellularity was approximately 20% in Group 1, but 70-80% in the hPTH treated groups. Transplantation of MNCs together with MSCs had no additional effect on bone marrow cellularity. However, the proportion of human CD13 and CD33 myeloid progenitor cells was higher in Group 3, while the proportion of human CD34 did not differ significantly between the three groups. The proportion of CXCR4 cells in Group 3 was larger than in Groups 1 and 2 but without statistical significance.

Conclusion

We have demonstrated a positive effect of hPTH on stem cell proliferation and a possible synergistic effect of MSCs and hPTH on the proportion of human hematopoietic progenitor cells, in a xenotransplantation model. Clinical trials of the use of hPTH after stem cell transplantation should be considered.     

髌骨血供及其临床意义

通过动脉灌注,对120侧不同年龄(新生儿～87岁)尸体的髌骨的血液供应进行了研宄。髌骨的动脉有髌主要动脉、下极动脉和髌底动脉,分别发自髌前动脉丛和髌周动脉环,成人在胃内互相吻合。对髌骨血管分布的临床意义进行了简要的讨论。     

T-Cell-Dependent Production of IgG by Human Cord Blood B Cells in Reconstituted SCID Mice

Reconstitution of severe combined immunodeficient (SCID) mice with human lymphocytes has recently allowed the elucidation of abnormalities of immune responses in various immunological disorders. In the present study, mononuclear cells (MNC) from neonatal cord blood and adult peripheral blood were intraperitoneally injected into SCID mice to examine induction of human Ig in respective mice recipients. Human IgG was consistently detected in the serum of SCID transferred with adult MNC, but only a few SCID recipients of cord blood MNC showed detectable but low levels of IgG in the serum. The combination experiments of isolated B and T cells disclosed that some interactions between B and T cells might be necessary for IgG production in transferred SCID mice. Notably, transfer of cord blood B cells with adult but not cord blood T cells resulted in efficient induction of IgG, associated with a change in subclass distribution. The results suggest that inability of neonatal B cells to produce IgG can be overcome by transfer with adult mature T cells into SCID mice.     

ILIB机理探讨

低能量激光血管内照射疗法治疗疾病是近年来发展起来的一种新的血液治疗技术，为经临床常规治疗效果不佳的患者提供一种新的辅助疗法。本文了ＩＬＩＢ的生物学作用机理，ＩＬＩＢ治疗仪的激光造反和使用方法及其临床应用。     

The Effect of Human Umbilical Cord Blood Cells on Survival and Cytokine Production by Post-Ischemic Astrocytes in Vitro

Cerebral ischemia induces death of all neural cell types within the region affected by the loss of blood flow. We have shown that administering human umbilical cord blood cells after a middle cerebral artery occlusion in rats significantly reduces infarct size, presumably by rescuing cells within the penumbra. In this study we examined whether the cord blood cells enhanced astrocyte survival in an in vitro model of hypoxia with reduced glucose availability. Primary astrocyte cultures were incubated for 2 h in no oxygen (95% N, 5% CO₂) and low glucose (1% compared to 4.5%) media. Cord blood mononuclear cells were added to half the cultures at the beginning of hypoxia. Astrocyte viability was determined using fluorescein diacetate/propidium iodide (FDA/PI) labeling and cytokine production by the astrocytes measured using ELISA. In some studies, T cells, B cells or monocytes/macrophages isolated from the cord blood mononuclear fraction with magnetic antibody cell sorting (MACS) were used instead to determine which cellular component of the cord blood mononuclear fraction was responsible for the observed effects. Co-culturing mononuclear cord blood cells with astrocytes during hypoxia stimulated production of IL-6 and IL-10 during hypoxia. The cord blood T cells decreased survival of the astrocytes after hypoxia but had no effect on the examined cytokines. Our data demonstrate that the tested cord blood fractions do not enhance astrocyte survival when delivered individually, suggesting there is either another cellular component that is neuroprotective or an interaction of all the cells is essential for protection.     

Effect of ABO Blood Group Incompatibility on the Outcome of Single-Unit Cord Blood Transplantation after Myeloablative Conditioning

ABO blood group incompatibility between donor and recipient has been associated with poor transplant outcomes in allogeneic hematopoietic stem cell transplantation. However, its effect on the outcome of cord blood transplantation (CBT) has yet to be clarified. We retrospectively analyzed 191 adult patients who received single-unit CBT after myeloablative conditioning for malignant disease in our institute. Major mismatch showed a significantly lower incidence of platelet engraftment compared with ABO match as a reference (hazard ratio, .57; P = .01). Nevertheless, there was no increase in graft-versus-host disease, transplant-related mortality, and overall mortality after ABO-incompatible CBT. These data suggested that donor–recipient ABO incompatibility does not have a significant impact on outcome after myeloablative CBT for hematological malignancies.     

Immunoregulatory Function of Human Cord Blood Lymphocytes on Immunoglobulin Production

ABSTRACT: The functional maturity of human umbilical cord blood B lymphocytes and the immunoregulatory activity of cord T lymphocytes were assessed by measuring the in vitro immunoglobulin production by B cells from either cord or adult blood. Supernatants from 48-hr pokeweed-mitogen (PWM) stimulated cord or adult lymphocyte cultures were added to cord or adult B cell cultures in the presence of PWM; a significant amount of immunoglobulin was produced in adult B cell cultures only. Adult B or T cells were then cocultured with cord T or B cells; a significant amount of immunoglobulin was again found only in adult B cell cultures. These results indicated that cord B cells were functionally immature and that cord helper T cell function was adequate but masked by excessive suppressor activity. Indeed, addition of cord T cells but not of allogeneic adult T cells to PWM stimulated adult lymphocyte cultures inhibited their immunoglobulin production; this confirmed cord T cells' increased suppressor activity. Cord T cells were not intrinsically suppressive since they failed to suppress immunoglobulin production by Epstein-Barr Virus (EBV) transformed B cells. They could be activated, however, by PWM or allogeneic cells (in mixed lymphocyte cultures) and their effect was mediated via soluble factor(s) as demonstrated by the suppressor effect of these culture supernatants on immunoglobulin production by unfractionated adult lymphocytes. In contrast, when these supernatants were added to T cell-depleted adult lymphyocyte cultures, enhancement rather than suppression was observed. These results indicated that the soluble factor(s) released by Cord T lymphocytes was not suppressing per se but induced suppression through activation of suppressor cells.     

Sustained Donor Engraftment in Recipients of Double-Unit Cord Blood Transplantation Is Possible Despite Donor-Specific Human Leukoctye Antigen Antibodies

The impact of human leukocyte antigen (HLA) donor-specific antibodies (DSA) on cord blood (CB) engraftment is controversial. We evaluated the influence of pre-existing HLA-antibodies (HLA-Abs) on engraftment in 82 double-unit CB recipients (median age, 48 years) who underwent transplantation for hematologic malignancies. Of 28 patients (34%) with HLA-Abs, 12 had DSA (median mean fluorescence intensity 5255; range, 1057 to 9453). DSA patients had acute leukemia (n = 11) or myelodysplasia (n = 1) and all received either high-dose or reduced-intensity (but myeloablative) conditioning. After myeloablative CB transplantation (CBT) (n = 67), sustained donor engraftment was observed in 95% without HLA-Abs (median, 23 days), 100% with nonspecific HLA-Abs (median, 23 days), and 92% with DSA (median, 31 days, P = .48). Of 6 patients with HLA-Abs to 1 unit, 3 engrafted with that unit and 3 with the other. Of 6 patients with HLA-Abs against both units, 1 had graft failure despite being 100% donor, and 5 engrafted with 1 unit. Successful donor engraftment is possible in patients with DSA after myeloablative double-unit CBT. Our data suggest potential deleterious effects of DSA can be abrogated in patients with hematologic malignancies.     

半月板的血供及其临床意义

通过动脉灌注,对56侧儿童和成人尸体的半月板血供进行了研究。成人半月板外1/5为血管区,内4/5为无血管区。半月板的撕裂常发生于无血管区或无血管区与血管区的连接处。半月板的退化和磨损位于无血管区。     

PUMA在大肠癌中的作用及临床意义

自PUMA发现以来，因具有强大的促凋亡功能而倍受瞩目，近年来对其分子结构、转录机制的研究取得了较大进展，其是否可以作为肿瘤治疗新靶点也一直备受关注，本文就其在大肠癌中研究进展做一简要综述。     

急性脊髓损伤后胶质酸性蛋白表达变化及意义

目的 观察急性脊髓损伤(SCI)后大鼠后肢功能恢复情况，以及损伤后胶质酸性蛋白(GFAP)表达的变化。方法 取雌性SD大鼠32只随机分为4组(n=8)：假手术组、伤后5d、9d、18d组．Allen法致伤脊髓．用大鼠综合行为评分法(CBS)对各级神经功能评分。用免疫荧光化学染色和图像分析的方法观察GFAP表达变化。结果 1．在行为观察中，发现大鼠脊髓损伤后有自行恢复倾向，损伤后18天后肢功能恢复67．5％。2．脊髓损伤后18d GFAP表达明显增强。结论 1、急性脊髓损伤后脊髓有自我修复的倾向。2、胶质细胞对脊髓损伤后的功能恢复起到重要作用。     

The distribution and characterization of skeletal muscle lesions in dysferlin-deficient SJL and A/J mice

The pathogenesis of limb-girdle muscular dystrophy type 2B (LGMD2B) dysferlinopathy remains to be investigated. The distribution and characterization of skeletal muscle lesions were examined in two different LGMD2B mouse models, SJL and A/J mice (at 10 and 35 weeks old), in association with the endoplasmic reticulum (ER) stress. SJL mice showed an earlier age of onset and a faster progression of skeletal muscle lesions as compared with those of A/J mice; the sensitivity difference to muscular dystrophic lesions between SJL and A/J mice was observed in the lumbar muscles (particularly, lumbar longissimus and sublumbar muscles); the lesions seen mainly in SJL mice at 35 weeks old consisted of degeneration, necrosis, fatty infiltration, variation in muscle fiber size and atrophy in muscle fibers. Enzyme-histochemically, the fast-twitch muscle fiber was predominant for the degenerative changes seen in the rectus femoris and lateral longissimus muscles of SJL mice. Immunohistochemically, the main reactive cell type observed in and around degenerative and/or necrotic muscle fibers was macrophages, demonstrable with an anti-F4/80 antibody. Because the analyses of spliced XBP1 mRNA, a marker of ER stress, did not show the increased expression, it was considered that ER stress did not affect the progression of skeletal muscle lesions in SJL mice with the advanced stage of dysferlinopathy.     

光量子血疗法治疗白塞氏综合征的疗效分析

采用光量子疗法（UBI）对22例白塞氏综合征（BS）病人进行治疗，以11例常规药物治疗的BS病人作为药物组对照疗效。同时检测二组病人治疗前后的超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH－Px）、肿瘤坏死因子（TNF）、脂质过氧化物（Lpo）、丙二醛（MDA）、过氧化氢酶（Cat）、免疫球蛋白、白细胞亚微结构等变化。结果表明，BS患者经UBI治疗后血中SOD、GSH－Px、Lpo有不同程度提高，而MDA、TNF有不同程度下降，白细胞微绒毛明显增多变粗。提示UBI对BS治疗效果优于常规药物，因而UBI是治疗BS的一种有效方法。     

Possible Role of Interleukin-10 in Autoantibody Production and in the Fate of Human Cord Blood CD5+ B Lymphocytes

Mononuclear cell subsets in 25 human umbilical cord blood samples and 10 healthy adults were studied. We found a decreased percentage of CD3+ cells, CD8+ cells and gammadelta T cells in cord blood compared with blood from healthy adults. The CD16+56+ and CD19+CD5+ phenotypes were overexpressed in cord blood. We then measured spontaneous gene expression and the production of interleukin-10 in mononuclear cells from cord blood and adult subjects. Although we found no difference between cord blood cells and those from healthy adults, a tendency towards spontaneous interleukin-10 production was observed in cord blood. Phorbol 12-myristate 13-acetate stimulation revealed that, among lymphocytes, cord blood B cells are the main cellular source of interleukin-10. Finally, we found no evidence of augmented spontaneous apoptosis but an increased bcl-2 gene expression in non-T cells from cord blood. Interleukin-10 might protect CD19+CD5+ B cells from apoptosis by inducing bcl-2 and promoting autoantibody production in this B-cell subpopulation.     

人外周血及脐血树突状细胞的体外分离培养

取正常人外周血或脐血,经淋巴细胞分离液分离,取中间白膜层,培养板中进行粘附,粘附细胞加培养液和细胞因子（外周血加GM－CSF和IL－4,脐血加GM－CSF和TNF－α）培养,对其形态、表型和功能分别进行鉴定和测定。结果表明约经过1周左右培养,悬浮细胞表现为典型的DC形态,带有毛刺样凸起,经DC单克隆抗体染色后用流式细胞仪测定脐血72％为DC,外周血93％为DC,并且可以刺激同种异体淋巴细胞的增殖反应。所以通过这样的不同细胞因子组合可以从人外周血和脐血中诱导培养出大量的DC细胞,为其进一步的研究奠定了基础。     

宫内窒息新生儿的脐血S100B蛋白检测的临床意义

目的:S100B蛋白是一种神经胶质特异性蛋白,可比较特异和灵敏地反映中枢神经系统的损伤.本文主要探讨宫内窒息新生儿脐血S100B蛋白的变化及对新生儿窒息诊断和窒息后脑损伤判断的价值.方法:采用正常阴道分娩组、剖宫产非窒息组和剖宫产宫内窒息组的新生儿脐血的S100B蛋白含量进行对比分析.结果:剖宫产宫内窒息组新生儿脐血的S100B蛋白含量明显高于其它两组,差异有显著性(P＜0.01);剖宫产非窒息组和正常阴道分娩组新生儿脐血的S100B蛋白含量差别无统计学意义(P＞0.05).结论:脐血S100B蛋白检测有助于新生儿窒息的诊断及窒息后脑损伤的判断.