COMPARISION OF THE APOPTOTIC EFFECTS ON LYMPHOBLASTS AND ON INCREASE OF MYELOID LINEAGE CELLS OF A SHORT-TIME,HIGH-DOSE METHYLPREDNISOLONE AND THE CONVENTIONAL-DOSE PREDNISOLONE TREATMENTS IN CHILDREN WITH ACUTE LYMPHOBLASTIC LEUKEMIA

The authors compare the apoptotic effect on the lymphoblasts and the proliferative effect on the myeloid lineage cells of a short-course high-dose methylprednisolone (HDMP) and the conventional-dose prednisolone treatments in children with acute lymphoblastic leukemia (ALL). The patients were divided into 2 groups. Group I (n = 10) received HDMP (30 mg/kg/day for 7 days) in a single dose before 6 a.m. perorally. Group II (n = 10) received prednisolone (2 mg/kg/day for 7 days) in 3 doses. The apoptotic percentages of lymphpblasts and the percentages of blasts and myeloid lineage cells were determined after performing the bone marrow aspiration (BMA) at diagnosis on the 0th, 3rd, and 7th days of the treatments in all patients. The mean apoptotic percentages of the lymphoblasts on the 3rd day were significantly higher than those on the 0th and 7th days in both groups (p <. 05). The highest apoptosis was determined on the 3rd day in group I. The mean percentages of the blast cells on the 7th day were significantly lower than those on the 0th and the 3rd days in both groups (p <. 05). The lowest lymphoblast percentage was determined on the 7th day in group I. The mean percentages of the CD₁₃⁺ and CD₃₃⁺ cells on the 7th day were significantly higher than those on the 0th and the 3rd days in both groups (p <. 05). The highest percentages of the CD₁₃⁺ and CD₃₃⁺ cells were found on the 7th day in group I. Prednisolone and HDMP showed no proliferative effect on the CD₁₄⁺ cells. These findings indicate that a short-course HDMP treatment shows a more effective apoptosis on the lymphoblasts and on the increase of the myeloid lineage cells when compared to the prednisolone treatment. The authors suggest that HDMP may be used in the treatment of patients with ALL instead of prednisolone.     

Proliferation of myeloid lineage cells and apoptosis of lymphoblastic leukemic cells induced by short-course high-dose methylprednisolone in patients with acute lymphoblastic leukemia

In this paper, we investigated the effects of short-course high-dose methylprednisolone (HDMP) treatment on the proliferation of myeloid lineage cells and on apoptosis of blast cells in eight children with acute lymphoblastic leukemia (ALL). The patients were given the HDMP treatment (30 mg/kg/d, perorally) before 9:00 a.m. for seven days. Bone marrow (BM) aspiration was done at days 0 and 3 of the HDMP treatment in all patients and at the 7th day of the HDMP treatment in six patients. Bone marrow blast cells had gradually decreased after the HDMP treatment by the 7th day. There were statistically significant differences between the mean percentages of BM blast cells at days 0 and 3, days 0 and 7, and at days 3 and 7 (p<0.05). The mean percentages of blast cell apoptosis at the 3rd day was significantly higher than at days 0 and 7, and apoptosis at day 0 was significantly lower than at the 7th day (p<0.05). The mean percentages of BM myeloid lineage cells at the 7th day was significantly higher than at days 0 and 3 (p<0.05), and the mean percentage at day 0 was significantly lower than at the 3rd day (p<0.05). These findings indicate that short-course HDMP treatment causes apoptosis on lymphoblasts and increases the proliferation of myeloid lineage cells in children with ALL.     

EFFECT OF SHORT-COURSE, HIGH-DOSE STEROID THERAPY IN A CHILD WITH MYELODYSPLASTIC SYNDROME

High-dose methylprednisolone (HDMP) has been shown to induce differentiation of myeloid leukemic cells with a remarkable antileukemic effect in children with various subtypes of acute myeloblastic leukemia (AML). Here the beneficial effect of short-course HDMP therapy in a child with myelodysplastic syndrome (MDS) is reported. Oral methylprednisolone sodium succinate (Prednol-L) was administered at a single daily dose of 30 mg/kg for 5 days to a 4-year-old girl with refractory anemia with excess of blasts and hypocellular bone marrow before the initiation of chemotherapy. In addition to dramatic clinical improvement, the patient's white blood cell count increased from 2.3 ×10 9 /L to 5.0 ×10 9 /L, and peripheral blood blast cells disappeared 4 days after HDMP treatment. Repeated bone marrow aspirate 1 week after the initiation of HDMP disclosed increased cellularity with no blasts. Furthermore, short-course HDMP treatment stimulated the increase in the number of peripheral blood lymphocytes and CD3 + , CD4 + , CD8 + , CD19 + , CD34 + , and NK cells. Results obtained with HDMP from the previous studies and the present case suggest that high-dose methylprednisoloneis a promising agent in the treatment of MDS and it is recommended as an initial treatment especially for MDS children with hypocellular bone marrow at presentation.     

EFFECT OF SHORT-COURSE,HIGH-DOSE STEROID THERAPY IN A CHILD WITH MYELODYSPLASTIC SYNDROME

High-dose methylprednisolone (HDMP) has been shown to induce differentiation of myeloid leukemic cells with a remarkable antileukemic effect in children with various subtypes of acute myeloblastic leukemia (AML). Here the beneficial effect of short-course HDMP therapy in a child with myelodysplastic syndrome (MDS) is reported. Oral methylprednisolone sodium succinate (Prednol-L) was administered at a single daily dose of 30 mg/kg for 5 days to a 4-year-old girl with refractory anemia with excess of blasts and hypocellular bone marrow before the initiation of chemotherapy. In addition to dramatic clinical improvement, the patient's white blood cell count increased from 2.3 &#50 10 9 /L to 5.0 &#50 10 9 /L, and peripheral blood blast cells disappeared 4 days after HDMP treatment. Repeated bone marrow aspirate 1 week after the initiation of HDMP disclosed increased cellularity with no blasts. Furthermore, short-course HDMP treatment stimulated the increase in the number of peripheral blood lymphocytes and CD3 + , CD4 + , CD8 + , CD19 + , CD34 + , and NK cells. Results obtained with HDMP from the previous studies and the present case suggest that high-dose methylprednisoloneis a promising agent in the treatment of MDS and it is recommended as an initial treatment especially for MDS children with hypocellular bone marrow at presentation.     

The effect of short-course high-dose methylprednisolone on peripheral blood CD34+ progenitor cells of children with acute leukemia during remission induction therapy

This study was undertaken to determine the effect of short-course high-dose methylprednisolone (HDMP) treatment on peripheral blood (PB) CD34+ progenitor cells during remission induction treatment in 11 children with newly diagnosed acute leukemia (7 with ALL, 4 with AML) whose bone marrow (BM) cells expressed fewer than 5% CD34 at the time of diagnosis. All children who had no infection were given HDMP as a single daily oral dose of 30 mg/kg for the first four days of induction therapy. The number of CD34+ progenitor cells were determined by flow cytometry before and after four days of HDMP treatment. While the number of PB blast cells significantly decreased after only a four-day course of HDMP treatment, the number of PB CD34+ progenitor cells increased in all patients. In addition, after four days of HDMP treatment polymorphonuclear leukocytes (PMN) and mononuclear cells (MNC) increased significantly (p < 0.05). We suggest that the potential beneficial effects of HDMP in the induction treatment of acute leukemia may occur partly by the stimulation of PB CD34+ hematopoietic progenitor cells in a short period of time.     

The role of short course of high-dose methylprednisolone in children with acute myeloblastic leukemia (FAB M2) presented with myeloid tumor

The authors have previously demonstrated a favorable effect of high-dose methylprednisolone (HDMP), which can induce differentiation and apoptosis of leukemic cells in children with acute myeloblastic leukemia (AML). Here, they evaluate the effect of short-course HDMP in 2 children with acute myeloblastic leukemia (AML-M2) presented with myeloid tumor (MT). Methylprednisolone (20 or 30 mg/kg/day) was given orally, in a single dose, without using other antileukemic agents. Rapid cytoreduction in MT, peripheral blood, and bone marrow blasts was observed in both children following short-course (4 or 7 days) HDMP treatment, possibly due to HDMP-induced differentiation and apoptosis of leukemic cells. The effects of HDMP should be explored in patients with other subtypes of AML who present with MT.     

TREATMENT OF KASABACH-MERRITT SYNDROME BY MEGADOSE METHYLPREDNISOLONE

The authors have previously demonstrated a favorable effect of high-dose methylprednisolone (HDMP), which can induce differentiation and apoptosis of leukemic cells in children with acute myeloblastic leukemia (AML). Here, they evaluate the effect of short-course HDMP in 2 children with acute myeloblastic leukemia (AML-M2) presented with myeloid tumor (MT). Methylprednisolone (20 or 30 mg/kg/day) was given orally, in a single dose, without using other antileukemic agents. Rapid cytoreduction in MT, peripheral blood, and bone marrow blasts was observed in both children following short-course (4 or 7 days) HDMP treatment, possibly due to HDMP-induced differentiation and apoptosis of leukemic cells. The effects of HDMP should be explored in patients with other subtypes of AML who present with MT.     

A NOVEL APPROACH TO TREATMENT WITH HIGH-DOSE STEROID AS A DIFFERENTIATION INDUCER IN CHILDREN WITH ACUTE MYELOBLASTIC LEUKEMIA

Several experimental studies have demonstrated that certain steroid hormones can induce differentiation of mouse myeloid leukemic cells to macrophages and granulocytes. We have shown that high-dose methylprednisolone treatment (HDMP, 20-30 mg/kg/day) can induce differentiation of leukemic cells to mature granulocytes in children with different morphological subtypes of acute myeloblastic leukemia (FAB AML M1, M2, M3, M4). In addition, apoptosis can also be induced in vivo and in vitro in AML blast by HDMP treatment. Short-course (3 to 5 days) HDMP treatment increases the hematopoietic CD34- positive progenitor cells in both bone marrow and peripheral blood in children with AML. Acceleration of leukocyte and neutrophil recovery has been obtained by the administration of short-course HDMP in chemotherapy-induced neutropenic children with AML. The addition of HDMP to anti leukemic chemotherapy increased the complete remission rate and prolonged the duration of remission in children with AML and significantly improved the outcome of AML children who presented with extramedullary infiltration. We suggest that the possibility of HDMP-induced differentiation and apoptosis should be evaluated in patients with other malignant diseases.     

High dose methylprednisolone therapy in nephrotic syndrome

This study was done to determine the efficacy of oral high dose methylprednisolone (HDMP) therapy in the treatment of childhood nephrotic syndrome (NS). Fifteen patients were enrolled in the study. Patients were arbitrarily divided into two groups. Group I received prednisolone (daily 60 mg/m² for 4 weeks, 45, 30, 20, 10, 5 mg/m² on alternate days for 4 weeks) and group II received HDMP (30 mg/kg/d for 3 days, 20 mg/kg/d for 4 days, 10 mg/kg/for a week, before 9 am, orally). The patients were followed-up for a duration of 38.0±5.5 months (range 24–68 months) in group I and 42.1±5.5 months (range 16–72 months) in group II. No significant difference was obtained in the duration of remission between both groups (p>0.05), while HDMP induced early remission than prednisolone (p<0.05). The mean relapse rate was 0.8/year in group I and 0.8/year in group II (p>0.05). Although, the number of the patients were limited in the study it can be recommended that patients with NS can be treated with oral HDMP therapy as an alternative to standard oral prednisolone therapy.     

In vitro determination of the apoptotic effect of heparin on lymphoblasts using DNA analysis and measurements of Fas and Bcl-2 proteins by flow cytometry

Heparin has an apoptotic effect beside its anticoagulant, anti-inflammatory, antihypertensive, and antiproliferative effects. In this study, the authors detected the percentages of apoptotic lymphoblasts and the expressions of apoptotic Fas protein and antiapoptotic Bcl-2 protein with flow cytometry in vitro after the incubation of lymphoblasts with heparin. Eleven newly diagnosed acute lymphoblastic leukemia (ALL) children were included in the study. Lymphoblasts were incubated in all different levels of heparin concentrations (0, 10, and 20 U/mL) and the percentages of apoptotic lymphoblasts and the percentages of Fas protein and Bcl-2 proteins were simultaneously measured by flow cytometry at 0, 1, and 2 h. At 0, 1, and 2 h, apoptosis was determined when heparin was added in 10- and 20-U/mL concentrations (p <.05). The apoptotic effect of heparin on lymphoblasts was higher at the first hour than at 0 and 2 h in 10- and 20-U/mL heparin concentrations (p <.01). The highest apoptosis was detected in the 20-U/mL heparin concentration at the first hour. The expression levels of Fas protein on lymphoblasts were higher at the first hour than at 0 and 2 h in 10- and 20-U/mL heparin concentrations (p <.001). The highest expression of Fas protein was observed in the 20-U/mL heparin concentration at the first hour. The expression levels of Bcl-2 protein on lymphoblasts were lower at the first hour than at 0 and 2 h in 10- and 20-U/mL heparin concentrations (p <.001). The lowest expression of Bcl-2 protein was detected in the 20-U/mL heparin concentration at the first hour. Increased concentrations of heparin had an increasing effect on the percentages of apoptotic lymphoblasts. The expression percentages of Fas protein on lymphoblasts also increased, whereas the expression percentages of Bcl-2 protein on lymphoblasts decreased (p <.05). These results suggest that low-dose heparin may cause significant apoptosis of lymphoblasts in newly diagnosed ALL patients.     

IL-15对儿童MDS造血前体细胞凋亡的影响

为探讨IL 1 5对儿童骨髓增生异常综合征 (MDS)造血细胞凋亡的影响 ,为临床应用提供理论依据。采用吸附单克隆抗体的免疫磁珠分离系统分离纯化 1 8例MDS患者骨髓CD34+细胞 ,采用碘化丙锭染色、流式细胞仪分析和细胞核形态检测两种方法同时检测体外培养的MDS造血前体细胞增殖、分化过程中的凋亡状况 ;观察IL 1 5对它们的影响 ,并进行了定量分析。结果表明 :发现IL 1 5可抑制体外培养的MDS造血前体细胞凋亡。在培养的第 8天 ,IL 1 51 0 0ng.ml- 1实验组细胞凋亡程度较没加IL 1 5的对照组明显减轻     

Organ-specific maturation of the major histocompatibility antigens in rats

The essential role of major histocompatibility complex (MHC) class I and II in the process of rejection has been documented, and some studies suggest that fetal transplants could enjoy an organ-specific immunologic privilege. However, little is known as to when these antigens develop in fetal organs and which tissues mainly present them. This study investigated the dynamics of immunogenicity in the developing transplant organs of rats. The study focused on the classic transplant organs including lung, heart, liver, pancreas, intestine, and kidney. Fetal organs (14th and 20th day of gestation), organs at the 3rd, 7th, 10th, 14th and 28th days postpartum (pp), 2 and 3 months pp, and adult organs were taken and snap-frozen in liquid nitrogen. MHC expression was analyzed applying the APAAP technique on serial cryosections by two well-defined monoclonal antibodies (mAb) generated against rat MHC class I (Ox 18) and class II (Ox 6). Immunoreactivities were compared to those of different monoclonal markers against endothelial cells (HIS52, CD 31), histiocytes (ED 1, ED 2), dendritic cells (Ox-62), granulocytes (HIS48), B-cells (RLN-9D3), T-cells (Ox-52), CD 4 (Ox-35), CD 8 (Ox-8a), natural killer cells (10/78), and CD 45 (Ox-1, leukocyte common antigen). A non specific mAb (MR 12/53) served as a negative control. In all stages of organ maturation, MHC I expression was found predominantly on immunocompetent cells, endothelial cells, and certain parenchymal cells, whereas MHC II was almost entirely restricted to dendritic cells. In organ development, the onset of MHC I expression and the number of MHC II-positive cells varied in a time-dependent manner. However, between the 2nd and 3rd month pp the expression pattern was comparable to adult organs. The study indicates that each organ carries a variable immunologic burden, that matures heterogeneously. Consequently, the variable content of MHC I/II in organ maturation needs to be considered for any transplantation model.     

卡介苗体外对急性淋巴细胞性白血病患儿细胞毒性T淋巴细胞杀伤HL-60细胞效应的影响

目的：探讨卡介苗（BCG）体外对急性淋巴细胞性白血病(ALL)患儿外周血细胞毒性T淋巴细胞（CTL）杀伤HL-60细胞效应的影响。方法：应用Ficoll-Hypaque法分离ALL患儿（白血病组）和健康儿童（对照组）的外周血单个核细胞,在白介素-2（IL-2）、植物血凝素（PHA）和BCG作用下诱导成CTL细胞;应用流式细胞仪测定CD3、CD3+CD4+、CD3+CD8+比例变化;应用MTT法检测CTL对HL-60细胞的杀伤力。结果：培养前2 d,对照组和白血病组CTL细胞数量及体积均无明显变化,第3天开始两组细胞均开始增殖,体积变大,6～10 d达高峰;培养至第10天时,加入BCG的白血病组和对照组细胞数量分别较无BCG的白血病组和对照组增多;白血病组CD3+CD8+比例明显高于对照组,经BCG作用后两组CD3+CD8+比例均明显升高;白血病组CTL对HL-60细胞的杀伤力明显低于对照组。结论：BCG可协同IL-2、PHA体外促进CTL细胞增殖并提高其对HL-60细胞的杀伤力。     

脂肪间充质干细胞的体外诱导和成血管作用

目的 研究在体外诱导脂肪间充质干细胞(ADMSCs)向内皮细胞分化、在立体培养基上的血管形成情况.方法 将传至第三代的ADMSCs用内皮细胞诱导液、Matrigel三维培养基进行诱导培养,对ADMSCs和诱导细胞选用CD31、CD44细胞表面抗原在流式细胞仪检测表达情况,用HE染色、FⅧ-RAg免疫组织化学染色及倒置显微镜、透射电镜等进行观察鉴定.结果流式细胞仪上检测ADMSCs的表达为CD44阳性、CD31阴性,诱导的细胞CD31阳性、CD44阴性.诱导细胞14d倒置显微镜下呈铺路石样形态,FⅧ-RAg染色细胞呈阳性,透射电镜下在细胞浆内见到内皮细胞特有标志物Weibel-Palade小体。ADMSCs在Matrigel三维培养基上诱导,24h细胞迁徙成团、有伪足伸出,诱导7d伸出的细胞形成交叉网格状,13d形成较长血管,20d较长血管粗而多、出现小分叉,30d长血管、分叉血管变粗厚;FⅧ-RAg染色后诱导血管亦呈阳性.结论 脂肪间充质干细胞在体外经诱导能向内皮细胞分化、能形成血管,可作为促进组织工程移植物血管化的良好的种子细胞.

Abstract：

Objective To study in vitro induction of adipose-derived mesenchymal stem cells (ADMSCs) into endothelial cells and blood vessel formation on three-dimensional (3D) media． Methods The 3rd passage ADMSCs were induced in vitro into endothelial cells on conditional medium and grew on Matrigel medium． The cell surface antigens CD31 and CD44 were detected with flow cytometric analysis before and after induction． HE staining, FⅧ-RAg immunohistochemical staining, inverted microscope and transmission electron microscope were used for morphological study.Results The expression of CD44 was positive and CD31 negative in ADMSCs in flow cytometric analysis.After induction,CD31 became positive while CD44 was negative． Paving-stone-like cell appearance was seen under inverted microscope 14 days after induction.The cells were FⅧ-RAg positively stained with immunohistological method,and Weibel-Palade body was observed under transmission electron microscope.On Matrigel media,the induced cells migrated in lumping with pseudopodia protrusion after 24 hours,and formed grid structure on the 7th day.Long vasculature was observed on the 13th day,and the vessels branched on the 20th to 30th day.The vessels stained positive for FⅧ-Rag． Conclusions ADMSCs have the potential to give rise to endothelial cells with in vitro induction and to form vessel structure in 3D media.ADMSCs have potential role in vascularized tissue engineering grafting．     

Lineage Switch in MLL‐Rearranged Infant Leukemia Following CD19‐Directed Therapy

Rearrangements of the mixed lineage leukemia (MLL) gene occur frequently in infants with both acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML). Conversions of leukemia cell lineage are rare, but occur most commonly in the setting of MLL‐rearrangement. Blinatumomab is a bidirectional antibody targeting CD19 with significant activity in relapsed B‐precursor ALL. We report an infant with ALL with t(4;11)(q21;q23) refractory to cytotoxic chemotherapy who was treated with blinatumomab. Following rapid initial clearance of peripheral lymphoblasts, bone marrow evaluation demonstrated a leukemic lineage switch to CD19‐negative monoblastic AML. Complete remission was achieved with myeloid‐directed chemotherapy.     

Comparison of oral alendronate versus prednisolone in treatment of infants with vitamin D intoxication

Aim: The aim of this report was to compare the efficacy of oral alendronate versus prednisolone treatment in addition to conventional measures in infants with vitamin D intoxication. Methods: In six infants (aged 8.0 ± 2.1 months) with vitamin D intoxication, time to achieve normocalcemia with prednisolone treatment (Group I, n = 4) or alendronate treatment (Group II, n = 4, two infants started treatment from the baseline and two after unsuccessful prednisolone treatment) in addition to intravenous hydration and diuretic therapy were compared . Results: Baseline serum calcium levels ranged between 3.8 and 4.77 mmol/L. In the prednisolone group, although two patients reached normocalcemia on 7th and 12th days of treatment, other two patients did not despite 23 and 15 days of treatment and therefore switched to alendronate treatment. The mean duration of prednisolone treatment in these four patients was 14.2 ± 6.7 days (range 7–23). In the alendronate group, two patients who started treatment from the baseline achieved normocalcemia on the 5th day. Other two patients achieved normocalcemia 2 days after switching to alendronate. Thus, the mean time to reach normocalcemia after single oral alendronate administration was 3.5 ± 1.7 days (range 2–5) (p < 0.01 versus Group I). Conclusion: Alendronate treatment achieves normocalcemia four times earlier than prednisolone treatment and shortens hospital stay in infants with vitamin D intoxication.     

小儿急性淋巴细胞白血病髓系抗原和CD_(34)表达的分析

为分析小儿急性淋巴细胞白血病 (ALL)的髓系抗原、CD34 表达特点 ,采用间接免疫荧光法分析 6 2例初治的ALL患者的免疫表型。结果①儿童ALL髓系抗原阳性率达 2 0 2 % ,其中CD13阳性最常见 (占 1 2 % )T系ALL和B系ALL髓系抗原表达差异无显著性。ALL L2 中的阳性率高于L1(P <0 0 5)。②CD34 表达阳性率为33 3% ,B ALL阳性率高于T ALL(P <0 0 1 ) ,B ALL中前B细胞型明显高于普通型、B细胞各亚型。③CD34 表达与外周血幼稚细胞有关 (P <0 0 1 )。结果表明儿童ALL髓质抗原表达和FAB分型有关 ;CD34 表达与外周血原始细胞和脑膜白血病的发生有关。与ALL亚型、细胞分化程度有关。     

三甲基化组蛋白H3赖氨酸4对调节性T淋巴细胞foxp3基因表达的影响

目的探讨天然调节性T淋巴细胞(nTreg)的三甲基化组蛋白H3赖氨酸4(H3K4me3)对foxp3基因的影响,寻找维持调节性T淋巴细胞(Treg)表型长期稳定的方法。方法使用免疫磁珠细胞(MACS)分选法从健康人外周血中分离出CD4+CD25+Treg。对CD4+CD25+Treg分别进行0 d、3 d、7 d培养,其中0 d为阴性对照组,3 d为阳性组1,7 d为阳性组2,采用流式细胞仪(FCM)对0 d、3 d、7 d nTreg膜表面标志CD25的变化进行检测,并分别对培养0 d、3 d、7 d的nTreg细胞通过染色质免疫共沉淀(ChIP-PCR)法检测foxp3基因启动子区H3K4me3及DNA水平变化。结果 1.使用细胞计数Kit-8(CCK-8)法确定植物血凝素(PHA)质量浓度在10 mg.L-1时为最佳的药物质量浓度,对Treg细胞的增殖作用最为显著。nTreg细胞培养0 d、3 d、7 d的表型变化呈递减趋势。随着培养时间(0 d、3 d、7 d)的延长,与foxp3基因启动子区结合的H3K4me3表达逐渐减少。2.采用ChIP-PCR法对培养0 d、3 d、7 d的nTreg细胞检测与H3K4me3结合的foxp3基因启动子区DNA水平变化。DNA凝胶电泳图显示,0 d、3 d在204 bp处可见条带(灰度值分别为2.31、0.91),7 d有模糊条带或无条带。三者比较差异有统计学意义(P<0.05)。结论 PHA不能维持Treg细胞的表型稳定,其机制与H3K4me3减少有关。     

胰岛素样生长因子-1经鼻给药对缺氧缺血性脑损伤新生大鼠内源性神经干细胞的影响

目的研究胰岛素样生长因子-1(IGF-1)经鼻靶向中枢给药对缺氧缺血性脑损伤(HIBD)新生大鼠内源性神经干细胞(NSCs)的保护性治疗作用。方法取90只7日龄SD新生大鼠随机分为假手术组、模型对照组、IGF-1干预组。采用Rice法制作新生大鼠HIBD动物模型。模型对照组于缺氧缺血后1 h鼻腔递送9 g·L-1盐水0.1 mL;IGF-1干预组于缺氧缺血后1 h鼻腔递送IGF-1 2.5μg(溶于9 g·L-1盐水0.1 mL);假手术组仅分离颈总动脉,不结扎不行缺氧处理。术后1、3、5、7、14 d取其脑组织,采用单、双标免疫组织化学方法检测其侧脑室室管膜下区(SVZ)BrdU阳性细胞和巢蛋白(Nestin)阳性细胞、双阳性细胞的表达;并计算BrdU阳性细胞数、Nestin阳性细胞数及BrdU-Nestin双阳性细胞数;HE染色观察其脑损伤组织形态结构变化。结果模型对照组:术后1 d SVZ区BrdU、Nestin、BrdU-Nestin阳性细胞开始增加,第3天达高峰,术后7 d明显减少,术后14 d SVZ仅有少量细胞;IGF-1干预组:术后1 d SVZ 3种阳性细胞明显增多,术后3 d增加急剧,术后5 d达高峰,术后7 d开始减少,术后14 d减少明显,每个时间点IGF-1干预组均高于模型对照组(Pa<0.05);假手术组无明显增殖现象,与其他2组比较差异有统计学意义(P<0.05)。结论经鼻腔导入IGF-1对HIBD大鼠内源性NSCs具有明显的保护作用。     

TPO和FL有效扩增脐血CD34+细胞为巨核前体细胞

目的探讨人脐血CD34+细胞在TPO和FL细胞因子组合作用下体外扩增为巨核前体细胞的效果,为开展血小板减少症的细胞治疗建立实验条件。方法本研究采用人脐血经免疫磁珠法纯化分离的CD34+细胞,研究TPO+FL细胞因子组合对体外巨核前体细胞的扩增效果。结果显示在TPO与FL组合的作用下,脐血CD34+细胞(n=6)显著地扩增为巨核前体细胞(CD61+CD41+细胞),作用14天为较理想时间。无论是全细胞数和CD34+细胞,或是CD61+CD41+细胞和CFUMK,经过体外扩增,均显著增加。结论研究表明脐血CD34+细胞在TPO和FL细胞因子组合作用下可有效地扩增为巨核前体细胞,第14天是培养较理想时间。