A Prognostic Model Based on Colony Stimulating Factors-related Genes in Triple-negative Breast Cancer

Objective Triple-negative breast cancer(TNBC)is the breast cancer subtype with the worst prognosis,and lacks effective therapeutic targets.Colony stimulating factors(CSFs)are cytokines that can regulate the production of blood cells and stimulate the growth and development of immune cells,playing an important role in the malignant progression of TNBC.This article aims to construct a novel prognostic model based on the expression of colony stimulating factors-related genes(CRGs),and analyze the sensitivity of TNBC patients to immunotherapy and drug therapy.Methods We downloaded CRGs from public databases and screened for differentially expressed CRGs between normal and TNBC tissues in the TCGA-BRCA database.Through LASSO Cox regression analysis,we constructed a prognostic model and stratified TNBC patients into high-risk and low-risk groups based on the colony stimulating factors-related genes risk score(CRRS).We further analyzed the correlation between CRRS and patient prognosis,clinical features,tumor microenvironment(TME)in both high-risk and low-risk groups,and evaluated the relationship between CRRS and sensitivity to immunotherapy and drug therapy.Results We identified 842 differentially expressed CRGs in breast cancer tissues of TNBC patients and selected 13 CRGs for constructing the prognostic model.Kaplan-Meier survival curves,time-dependent receiver operating characteristic curves,and other analyses confirmed that TNBC patients with high CRRS had shorter overall survival,and the predictive ability of CRRS prognostic model was further validated using the GEO dataset.Nomogram combining clinical features confirmed that CRRS was an independent factor for the prognosis of TNBC patients.Moreover,patients in the high-risk group had lower levels of immune infiltration in the TME and were sensitive to chemotherapeutic drugs such as 5-fluorouracil,ipatasertib,and paclitaxel.Conclusion We have developed a CRRS-based prognostic model composed of 13 differentially expressed CRGs,which may serve as a useful tool for predicting the prognosis of TNBC patients and guiding clinical treatment.Moreover,the key genes within this model may represent potential molecular targets for future therapies of TNBC.

Molecular cloning, pathologically-correlated expression and functional characterization of the colony- stimulating factor 1 receptor （CSF-1R） gene from a teleost, Plecoglossus altivelis

Colony-stimulating factor 1 receptor （CSF-1R） is an important regulator of monocytes/macrophages （MO/MФ）. Although several CSF-1R genes have been identified in teleosts, the precise role of CSF- 1R in ayu （Plecoglossus altivelis） remains unclear. In this study, we characterized the CSF-1R homologue from P. altivelis, and named it PaCSF-1R. Multiple sequence alignment and phylogenetic tree analysis showed that PaCSF-1R was most closely related to that of Japanese ricefish （Oryzias latipes）. Tissue distribution and expression analysis showed that the PaCSF-1R transcript was mainly expressed in the head kidney-derived MO/MФ, spleen, and head kidney, and its expression was significantly altered in various tissues upon Vibrio anguillarum infection. After PaCSF-1R neutralization for 48 h, the phagocytic activity of MO/MФ was significantly decreased, suggesting that PaCSF-1R plays a role in regulating the phagocytic function of ayu MO/M（P.

Application of pegylated recombinant human granulocyte colony-stimulating factor(PEG-rhG-CSF) for the prevention of neutropenia in triple negative breast cancer patients older than 65 years during adjuvant chemotherapy

Objective The aim of this study was to compare the efficacy and safety of pegylated recombinant human granulocyte colony-stimulating factor(PEG-rhG-CSF)and recombinant human granulocyte colonystimulating factor(rhG-CSF)for the prevention of neutropenia in elderly breast cancer patients during adjuvant chemotherapy.Methods A total of 45 oncology inpatients with breast cancer,who received adjuvant chemotherapy and were older than 65 years from May 2017 to October 2018 in the General Hospital of the Northern Theater of the Chinese people’s Liberation Army,were included.Epirubivin Cyclophoshamide-Docetaxel(EC-T)sequential adjuvant chemotherapy was chosen.Forty-five patients were randomly divided into two groups;25 patients in the treatment group were treated with PEG-rhG-CSF and 20 patients in the control group were not treated with PEG-rhG-CSF,but only rhG-CSF.The experimental group was treated with the PEG-rhG-CSF at the end of chemotherapy for 24–48 h,with a 6 mg subcutaneous injection once per chemotherapy cycle.In the control group,rhG-CSF was administered after 48 h of chemotherapy,with a 100μg subcutaneous injection,1/d,d 1–7.The dosage could be increased step by step with the exacerbation of neutropenia.The primary aims of this study was to discover the incidence of leukopenia,neutropenia,neutrophilic fever,and adverse reactions in the two groups.Results The incidence of neutropenia,neutrophilic fever and adverse reactions decreased in the treatment group compared to the control group,but no significant difference existed between two groups(P>0.05).Patients in treatment group had a lower,but not statistically significant,incidence of adverse reactions(P>0.05).Conclusion Applying PEG-rhG-CSF could be effective in preventing neutropenia in elderly patients with postoperative adjuvant chemotherapy to treat breast cancer.It may effectively control the occurrence of neutropenia after chemotherapy and reduce the chance of infection.The incidence of side effects,such as fever and bone pain,was low.The adverse drug reactions were well tolerated by patients,which could ensure the smooth progress of chemotherapy.

rh-CSF1改善缺糖缺氧损伤神经元线粒体功能和细胞凋亡

目的探讨集落刺激因子-1(colony stimulating factor-1,CSF1)抑制氧糖剥夺(oxygen-glucose deprivation,OGD)神经元凋亡的作用机制。方法采用大鼠原代大脑皮质神经元,分为OGD损伤神经元模型组(OGD组,n=3)、重组人CSF1(recombined human CSF1,rh-CSF1)干预组(rh-CSF1组,n=3)、对照组(n=3)。测定3组神经元凋亡率和其中三磷酸腺苷(adenosine triphosphate,ATP)含量,活性氧簇水平、线粒体膜电位和线粒体脱氧核糖核酸(deoxyribonucleic acid,DNA)拷贝数,检测线粒体内丙二醛含量和超氧化物歧化酶活性。结果OGD组模型进行基线评估,结果示神经元凋亡率、活性氧簇、线粒体内丙二醛水平、线粒体膜电位、线粒体DNA拷贝数、ATP含量、线粒体内超氧化物歧化酶活性与对照组有统计学差异(P<0.01)。rh-CSF1干预可提高OGD损伤后神经元的线粒体膜电位(0.55±0.03 vs.0.43±0.06,P<0.01)、线粒体DNA拷贝数(0.88±0.05 vs.0.72±0.06,P<0.05)、ATP含量([15.70±0.99)mmol/mg vs(.11.70±1.00)mmol/mg,P<0.01)]和线粒体内超氧化物歧化酶活性([18.47±1.38)U/mg vs.14.78±1.81)U/mg,P<0.05)],降低活性氧簇(3.64±0.21 vs.4.45±0.33,P<0.05)和线粒体内丙二醛水平([2.13±0.19)mmol/mg vs(.2.78±0.20)mmol/mg,P<0.05)],减轻神经元凋亡率。结论rh-CSF1可能通过改善线粒体功能、减轻氧化应激及抑制细胞凋亡,从而改善OGD诱导损伤神经元的受损程度。

rhG-CSF在治疗儿童化疗性口腔黏膜炎中的应用研究

目的探究重组人粒细胞集落刺激因子(recombinant human granulocyte-colony stimulating factor,rhG-CSF)在治疗儿童化疗性口腔黏膜炎中的应用效果。方法选取2020年1月至2022年6月于笔者医院肿瘤外科化疗的60例化疗性口腔黏膜炎患儿,按随机数字表法分为两组,每组30例。对照组采取生理盐水口腔护理,实验组采取rhG-CSF口腔护理。比较两组患儿干预效果、口腔黏膜炎分度、生存质量[健康状况调查简表(36-item short—form,SF-36)]、患儿家长护理满意度。结果实验组总有效率较对照组高,差异有统计学意义(P<0.05)。干预5d,实验组患儿口腔黏膜炎分度结果优于对照组,差异有统计学意义(P<0.05)。干预5d,两组患儿SF-36评分较干预前高,实验组较对照组高,差异有统计学意义(P<0.05)。实验组患儿家长满意度较对照组高,差异有统计学意义(P<0.05)。结论rhG-CSF口腔护理可有效提高儿童化疗性口腔黏膜炎的干预效果,减轻口腔黏膜炎分度,改善患儿生存质量,提高患儿家长护理满意度。

IMMUNOHISTOCHEMICAL OBSERVATION OF MACROPHAGE COLONY STIMULATING FACTOR AND ITS RECEPTOR IN BREAST CANCER AND HEPATOMA TISSUES

Objective: To study the potential role of cellular macrophage colony-stimulating factor (cM-CSF) and cellular macrophage colony-stimulating factor receptor (cM-CSF-R) with breast cancer and hepatoma and search the way for clinical application. Methods: Frozen surgical specimens from 48 breast cancer patients, including 29 cases of histological grade II and 19 eases of grade III, and 16 hepatoma patients were investigated by Avidin Biotin Complex (ABC) immunohistochemical assay with anti-M-CSF monoclonal antibody (Mab) and anti-M-CSF-R Mab. Pathohistological examination was performed as well. Results: cM-CSF and cM-CSF-R were detected in tested specimens. The expression levels of cM-CSF and cM-CSF-R in grade III group were higher than in grade II group and more higher than control group hyperplasia of breast. Hepatoma tissues also showed higher expression level of cM-CSF and cM-CSF-R than normal adult and fetal liver. Conclusion: Breast cancer and hepatoma tissues presented higher expression levels of cM-CSF and cM-CSF-R than control and expression level might be related with tumor’s process.

Granulocyte colony-stimulating factor-producing hepatocellular carcinoma with abrupt changes

Granulocyte colony-stimulating factor(G-CSF)-producing tumor is one of the rare types of cancer clinically characterized by an elevated fever and white blood cell(WBC) increment. Although G-CSF producing tumors have been reported in several types of cancer including those of the lungs, cervix and bladder, G-CSF producing hepatocellular carcinoma is extremely rare. Here, we report the case of a rapidly growing and poorly differentiated hepatocellular carcinoma producing G-CSF. The patient showed symptoms of continuous high fever, stomach pain and cough, and high serum WBC counts, C-reactive protein(CRP) and G-CSF levels were found in laboratory tests. After a radical hepatectomy, the patient completely recovered from the above symptoms and inflammatory state. The serum levels of G-CSF were reduced to normal levels after radical surgery. An immunohistochemical analysis revealed the overexpression of G-CSF in the cytoplasm of certain hepatocellular carcinoma(HCC) cell. The patient's serum WBC, CRP and G-CSF levels remained within normal levels in the six months after surgery without recurrence. This is the 9^(th) case report of G-CSF producing hepatocellular carcinoma in English literature. We review the clinical characteristics of the G-CSF producing HCC and discuss a possible treatment strategy.

ANTIGEN ASSOCIATION OF J6-1 CELL MEMBRANE ASSOCIATEDFACTOR RECEPTOR WITH MACROPHAGE COLONYSTIMULATING FACTOR RECEPTOR

Objective: To verify the antigen association of MAF-J6-1 receptor with M-CSFR and to further study the role of M-CSF and its receptor mediated juxtacrine in promoting leukemic cell proliferation. Methods: Monoclonal antibody (McAb) of MAF-J6-1R RE2 and polyclonal antibody (PolyAb) of rhM-CSFR were prepared. The specificity of McAb RE2 to M-CSFR was confirmed by indirect ELISA, cross-neutralizing assay with J6-1 cell colony formation and neutralization test by ELISA. Results: the reactive activity of purified RE2 to M-CSFR was over 1: 16000. The inhibitory activity of M-CSFR and MAF-J6-1R could be blocked by RE2 and anti-M-CSFR antibody. The reactivity of RE2 to M-CSFR could be reduced by M-CSFR. Conclusion: The specificity of RE2 to M-CSFR was confirmed and the antigen association of MAF-J6-1R with M-CSFR was proved. It suggests that M-CSF and its receptor mediated auto-juxtacrine stimulation could be an operative mechanism in either leukemia or nonhematological malignancies.

Granulocyte-colony stimulating factor therapy improves survival in patients with hepatitis B virus-associated acuteon-chronic liver failure

AIM:To evaluate the safety and efficacy of granulocyte-colony stimulating factor(G-CSF) therapy in patients with hepatitis B virus(HBV)-associated acuteon-chronic liver failure(ACLF).METHODS:Fifty-five patients with HBV-associated ACLF were randomized into two groups:the treatment group and the control group.Twenty-seven patients in the treatment group received G-CSF(5 μg/kg per day,six doses) treatment plus standard therapy,and 28 patients in the control group received standard therapy only.The peripheral CD34 + cell count was measured consecutively by flow cytometry.Circulating white blood cell count,biochemical parameters,and other clinical data of these patients were recorded and analyzed.All patients were followed up for a period of 3 mo to evaluate the changes in liver function and survival rate.RESULTS:The peripheral neutrophil and CD34 + cell counts in the G-CSF group increased on day 3 from the onset of therapy,continued to rise on day 7,and remained elevated on day 15 compared to those of the control group.Child-Turcotte-Pugh score of patients in the treatment group was improved on day 30 from the onset of G-CSF therapy,compared to that in the controls(P = 0.041).Model for End-Stage of Liver Disease score of patients in the treatment group was improved on day 7(P = 0.004) and remained high on day 30 from the onset of G-CSF therapy(P < 0.001) compared to that in controls.After 3 mo of follow-up observation,the survival rate in the treatment group(48.1%) was significantly higher than that in the control group(21.4%)(P = 0.0181).CONCLUSION:G-CSF therapy promoted CD34 + cell mobilization in patients with HBV-associated ACLF,and improved the liver function and the survival rate of these patients.

CO-EXPRESSION OF MACROPHAGE COLONY-STIMULATING FACTOR WITH ITS RECEPTOR IN HUMAN HEPATOMA CELLS AND ITS POTENTIAL ROLES

Objective: To investigate the potential role of macrophage colony-stimulating factor (M-CSF) and macrophage colony-stimulating factor receptor (M-CSF-R) on the growth of human hepatoma cells. Methods: Specimens of different origin, including tissues of human hepatocellular carcinoma (HCC), human fetal liver (FL) and normal liver (NL), the hepatoma cell lines, as well as the peripheral blood mononuclear cells (PBMC) from patients with HCC or liver metastatic tumor (LMT), were used to detect the expression levels of M-CSF and M-CSF-R by ABC immunohistochemistry staining and reverse transcription polymerase chain reaction methods the expression levels of M-CSF and M-CSF-R. Influence of monoclonal antibody against M-CSF (B5) or M-CSF-R (RE2) on proliferation ability of hepatoma cell linesin vitro was also studied. Results: The results showed that hepatoma tissues produced elevated levels of both M-CSF and M-CSF-R compared with those of fetal liver (P<0.001). The M-CSF/M-CSF-R expression levels of PBMC from hepatoma patients were higher than those of LMT patients (P<0.01,P<0.05) and the normal people (P<0.001). The hepatoma cell lines showed strong positive for M-CSF and M-CSF-R production. Both B5 and RE2 displayed a dose-dependent inhibitory effect on the growth and proliferation of hepatoma cells. Conclusion: The study indicates a co-expression model for M-CSF-R in hepatoma cells, suggesting an involvement of M-CSF/M-CSF-R in growth signaling of those malignant cells. The M-CSF/M-CSF-R seems to function through an autonomy mechanism in human hepatoma.

可视化分析GM-CSF在免疫炎症中的研究现状

目的通过文献计量学方法对粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage colony stimulating factor,GM-CSF)在免疫炎症方面相关研究现状、热点及发展趋势进行分析。方法在Web of Science核心数据库中检索1990年1月1日至2024年1月1日的相关文献,并应用CiteSpace软件对数据进行可视化分析。结果共纳入1219篇GM-CSF在免疫炎症方面相关文献,发文量整体呈上升趋势,美国以445篇居全球第一。发文量最高的机构是墨尔本大学25篇;发文量并列第一的作者是Jordana M和Becher B,每人10篇,被引次数最高的作者是Hamilton JA 128次;被引次数最多的期刊是Journal of Immunology 986次,GM-CSF在免疫炎症中相关热点主要包括inflammation、dendritic cell、T cell等,近几年的研究热点集中在immunity和microglia。结论随着GM-CSF在免疫炎症领域研究不断深入,其学术影响力也逐渐广泛,未来研究方向在于探索GM-CSF在免疫炎症领域中的作用机制和靶向治疗。

PEG-rhG-CSF与rhG-CSF预防肿瘤化疗相关中性粒细胞减少的有效性和安全性比较的系统评价和Meta分析

目的:比较聚乙二醇化重组人粒细胞集落刺激因子(PEG-rhG-CSF)与重组人粒细胞集落刺激因子(rhG-CSF)预防肿瘤化疗相关中性粒细胞减少的有效性和安全性。方法:检索PubMed、Embase、the Cochrane Library、中国知网、万方数据库和维普数据库,纳入PEG-rhG-CSF与rhG-CSF预防肿瘤化疗相关中性粒细胞减少(CIN)的随机对照试验(干预措施为使用PEG-rhG-CSF;对照措施为使用rhG-CSF),检索时间截至2023年5月。文献筛选和信息提取由2名研究者独立完成,并进行偏倚风险评估。采用RevMan 5.3统计软件对提取的数据进行Meta分析。结果:共纳入41篇文献。Meta分析结果显示,与rhG-CSF比较,PEG-rhG-CSF可以显著降低化疗后中性粒细胞减少伴发热(FN)发生率(OR=0.64,95%CI=0.51~0.81,P=0.0002)和Ⅲ/Ⅳ度中性粒细胞减少发生率(OR=0.49,95%CI=0.35~0.69,P<0.0001),差异均有统计学意义,但Ⅲ/Ⅳ度中性粒细胞减少持续时间(MD=-0.33,95%CI=-0.68~0.01,P=0.06)、中性粒细胞减少恢复时间(MD=-0.11,95%CI=-0.32~0.09,P=0.27)、骨痛或骨骼肌肉痛发生率(OR=0.81,95%CI=0.66~1.00,P=0.05)的差异均无统计学意义。结论:PEG-rhG-CSF预防化疗后FN及Ⅲ/Ⅳ度中性粒细胞减少的有效性明显优于rhG-CSF,PEG-rhG-CSF作为一级预防具有可行性。

Mobilization of Peripheral Blood Stem Cells Using Regimen Combining Docetaxel with Granulocyte Colony-stimulating Factor in Breast Cancer Patients

Objective：To evaluate the effectiveness and safety of the mobilization of peripheral blood hematopoietic stem cells by combining docetaxel with granulocyte colony-stimulating factor（G-CSF） in breast cancer patients.Methods：A total of 57 breast cancer patients were treated with docetaxel 120 mg/m2.When the white blood cell（WBC） count decreased to 1.0×109/L,patients were given G-CSF 5-g/kg daily by subcutaneous injection until the end of apheresis.Peripheral blood mononuclear cells（MNC） were isolated by Cobe Spectra Apheresis System.The percentage of CD34＋ cell was assayed by flow cytometry.Results：At a median 6 of days（range 3-8） after the administration of docetaxel,the median WBC count decreased to 1.08×109/L（range 0.20-2.31）.The median duration of G-CSF mobilization was 3 days（range 2-7）.The MNC collection was conducted 8-12 days（median 10 days） after docetaxel treatment.The median MNC was 5.35×108/kg（range 0.59-14.07）,the median CD34＋ cell count was 2.43×106/kg（range 0.16-16.69）.The CD34＋ cell count was higher than 1.00×106/kg in 47 of 57 cases（82.46%） and higher than 2.00×106/kg in 36 cases（63.16%）.The CD34＋ cell count was higher than 2.00×106/kg in 27 collections（23.68%）.The MNC count and the CD34＋ cell count were correlated with the bottom of WBC after docetaxel chemotherapy（r=0.364,0.502,P=0.005,0.000）.The CD34＋ cell count was correlated with the MNC count（r=0.597,P=0.000）.The mobilization and apheresis were well tolerated in all patients.Mild perioral numbness and numbness of hand or feet were observed in 3 cases.No serious adverse events were reported.Conclusion：Mobilization of peripheral blood hematopoietic stem cell by combining docetaxel with G-CSF was effective and safety in breast cancer patients.

G-CSF介导的小胶质细胞自噬活性降低在大鼠酒精使用障碍脑卒中中的脑保护作用研究

目的探讨大鼠酒精使用障碍(AUD)脑卒中后粒细胞集落刺激因子(G-CSF)治疗对小胶质细胞自噬活性的影响及其神经保护作用。方法将54只SD雄性大鼠按照随机数字表法分为AUD组、AUD+大脑中动脉栓塞(MCAO)组、AUD+MCAO+G-CSF组,每组各18只。AUD组用20%酒精双瓶法构建AUD大鼠模型,AUD+MCAO组、AUD+MCAO+G-CSF组均在AUD模型基础上制作MCAO局部脑缺血模型,AUD+MCAO+G-CSF组大鼠缺血再灌注损伤1 h后腹腔注射G-CSF 50μg/kg治疗。每天记录所有大鼠的酒精摄入量及偏好。观察各组大鼠脑缺血再灌注损伤24 h后检测脑梗死面积和神经功能评分。采用免疫荧光染色及蛋白质印迹法检测各组大鼠缺血侧大脑皮层小胶质细胞特异性标志物TMEM119、自噬标记物LC-3的表达水平。结果到末次饮酒28 d时酒精摄入量已达到稳定水平,酒精偏好明显增加。与AUD组比较,AUD+MCAO组的脑梗死面积显著升高,神经功能评分显著减少,差异均有统计学意义(P<0.05);与AUD+MCAO组比较,AUD+MCAO+G-CSF组的脑梗死面积明显减少,神经功能评分明显升高,差异均有统计学意义(P<0.05)。免疫荧光染色结果显示:与AUD组比较,AUD+MCAO组TMEM119和LC-3蛋白表达荧光强度明显增强,差异均有统计学意义(P<0.05);与AUD+MCAO组比较,AUD+MCAO+G-CSF组TMEM119和LC-3蛋白表达荧光强度均明显减弱,差异均有统计学意义(P<0.05)。此外,免疫荧光显示TMEM119和LC-3二者有明显共定位。蛋白质印迹法结果显示,TMEM119和LC-3在AUD组表达微弱,与AUD组比较,AUD+MCAO组TMEM119和LC-3蛋白表达均明显升高,差异均有统计学意义(P<0.05);与AUD+MCAO组比较,AUD+MCAO+G-CSF组TMEM119和LC-3蛋白表达明显降低,差异均有统计学意义(P<0.05)。结论G-CSF通过降低小胶质细胞的自噬活性,在AUD脑损伤中发挥重要的脑保护作用。

CSF1R基因突变致ALSP发病研究进展

成人发病的白质脑病合并轴索球样变和色素性胶质细胞(adult-onset leukoencephalopathy with axonal spheroids and pigmented glia,ALSP)是临床罕见的常染色体显性遗传病,其具体的发病机制目前还未明确。集落刺激因子1受体(colony-stimulating factor 1 receptor,CSF1R)是一种细胞表面跨膜酪氨酸激酶受体,与其相关的编码基因突变已被证实是ALSP的潜在致病因素。然而,目前关于CSF1R基因突变致使ALSP发病的具体机制尚不清楚。本文回顾CSF1R基因在ALSP发病过程中的突变位点及致病机制研究,发现CSF1R突变可以通过显性负性效应、功能丧失、单倍体剂量不足及功能获得等机制导致小胶质细胞功能异常,进而引起ALSP的发病。对ALSP病因的深入认识有助于更好地探索潜在的治疗方法。

PEG-rhG-CSF在血液肿瘤自体造血干细胞动员中的临床分析

目的:探讨无外周血CD34监测下聚乙二醇重组人粒细胞集落刺激因子(PEG-rhG-CSF)在血液肿瘤自体外周血干细胞动员中的采集时机及采集效果。方法:回顾性分析2017年8月至2022年1月福建医科大学附属第一医院收治的46例行自体外周血干细胞动员的血液恶性肿瘤患者。采用大剂量化疗联合PEG-rhG-CSF或重组人粒细胞集落刺激因子(G-CSF)动员方案,其中应用PEG-rhG-CSF动员的27例(PEG-rhG-CSF组),应用G-CSF动员的19例(G-CSF组),比较两组患者动员采集效果。结果:46例患者共采集86例次,PEG-rhG-CSF组与G-CSF组获得采集物的单个核细胞中位数分别为6.54(3.85-12.61)×10^(8)/kg和6.15(1.13-11.58)×10^(8)/kg(P>0.05),采集物CD34^(+)细胞数分别为11.44(1.33-65.02)×10^(6)/kg和4.95(0.30-24.02)×10^(6)/kg(P<0.05),采集时机分别为14(10-20)和14(4-22)d(P>0.05)。PEG-rhG-CSF组在外周血白细胞(WBC)≥10×10^(9)/L时单次所采集的产物CD34^(+)细胞数明显高于外周血WBC<10×10^(9)/L时采集的数量[19.04(2.85-65.02)×10^(6)/kg vs 6.22(0.81-34.86)×10^(6)/kg,(P<0.05)]。结论:采用PEG-rhG-CSF动员外周血干细胞单次采集足量CD34^(+)细胞成功率高,中位动员时间为14 d;在无外周血CD34监测情况下,外周血WBC≥10×10^(9)/L可以考虑作为单次采集足量干细胞的采集阈值。

Influence of granulocyte-macrophage colonystimulating factor and tumor necrosis factor on anti-hepatoma activities of human dendritic cells

INTRODUCTIONDendritic cells (DCs) play a key regulatory role inantitumor immunity,especially in its immuneaccessory role via MHC-Ⅰ molecules.We haverecently reported that DCs were able to enhance thekilling activity of Lymphokine and PHA activatedkiller (LPAK) cells in vitro.In the presentstudy,we evaluated the effects of GM-CSF andTNF upon antitumor activities of freshly

Neuroprotective Effect of Granulocyte Colony-stimulating Factor in a Focal Cerebral Ischemic Rat Model with Hyperlipidemia

Granulocyte colony-stimulating factor (G-CSF) has been demonstrated to have neuroprotective effects in rat model with focal cerebral ischemia through anti-apoptotic pathways and by promoting proliferation of neural stem cells. In the present study, we examined the neuroprotective effect of G-CSF in an acute focal cerebral ischemia rat model with lipid metabolism disorder. Eighty male SD rats were randomly divided into normal diet control group (NC group) and high-fat diet group (HFD group) (n = 40 in each). In HFD group, rats were fed on high fat diet to induce atherosclerosis. After 29 days, 4 rats from each group were sacrificed to evaluate the effects of different diets, and the middle cerebral artery occlusion (MCAO) was performed in the rest of the rats. MCAO rats received either G-CSF (50 μg·kg–1·mL–1) or phosphate buffered saline (PBS) injection through the external jugular vein for 5 days, which was followed by 5-bromo-deoxy uridine (BrdU, i.p., 50 mg/kg) injection for another 7 days. To evaluate the effects of G-CSF treatment on neurological function, the modified neurological severity score (mNSS) was calculated. The vascular distribution, ischemic cells proliferation, cell apoptosis and the expression of vascular endothelial growth factor (VEGF) were measured to determine the effects of G-CSF treatment. Our results showed that G-CSF-treated rats had a lower mNSS than PBS-treated rats in both NC group and HFD group. G-CSF injection promoted endothelial cell proliferation and vascular regeneration, and inhibited cell apoptosis. The serum and tissue levels of VEGF were significantly increased after G-CSF treatment. It is concluded that G-CSF exerts its neuroprotective effect in focal cerebral ischemia rats with hyperlipidemia by enhancing angiogenesis, promoting cells proliferation, decreasing cell apoptosis, and increasing local VEGF expression.

Impaired granulocyte-macrophage colony-stimulating factor bioactivity accelerates surgical recurrence in ileal crohn's disease

AIM To examine the relationship between elevated granulocyte-macrophage colony-stimulating factor(GMCSF) auto-antibodies(Ab) level and time to surgical recurrence after initial surgery for Crohn's disease(CD). METHODS We reviewed 412 charts from a clinical database at tertiary academic hospital. Patients included in the study had ileal or ileocolonic CD and surgical resection of small bowel or ileocecal region for management of disease. Serum samples were analyzed for serological assays including GM-CSF cytokine, GM-CSF Ab, ASCA Ig G and Ig A, and genetic markers including SNPs rs2066843, rs2066844, rs2066845, rs2076756 and rs2066847 in NOD2, rs2241880 in ATG16 L1, and rs13361189 in IRGM. Cox proportional-hazards models were used to assess the predictors of surgical recurrence.RESULTS Ninety six percent of patients underwent initial ileocecal resection(ICR) or ileal resection(IR) and subsequently 40% of patients required a second ICR/IR for CD. GMCSF Ab level was elevated at a median of 3.81 mcg/mL. Factors predicting faster time to a second surgery included elevated GM-CSF Ab [hazard ratio(HR) 3.52, 95%CI: 1.45-8.53, P = 0.005] and elevated GM-CSF cytokine(HR = 2.48, 95%CI: 1.31-4.70, P = 0.005). Factors predicting longer duration between first and second surgery included use of Immunomodulators(HR = 0.49, 95%CI: 0.31-0.77, P = 0.002), the interaction effect of low GM-CSF Ab levels and smoking(HR = 0.60, 95%CI: 0.45-0.81, P = 0.001) and the interaction effect of low GM-CSF cytokine levels and ATG16 L1(HR = 0.65, 95%CI: 0.49-0.88, P = 0.006).CONCLUSION GM-CSF bioavailability plays a critical role in maintaining intestinal homeostasis. Decreased bioavailability coupled with the genetic risk markers and/or smoking results in aggressive CD behavior.

CONSTRUCTION OF EUKARYOTIC EXPRESSION VECTOR WITH GRANULOCYTE-MACROPHAGE COLONY-STIMULATING FACTOR GENE

Objective: To construct the eukaryotic expression vector that express human granulocyte-macrophage colony-stimulating factor (hGM-CSF) gene for making highly express in mammalian cells. Methods: Extract totally RNA from the induced human fetal lung (HFL) cell line. HGM-CSF cDNA was obtained by reverse transcription-polymerase chain reaction (RT-PCR), and then directionally subcloned into the HindIII and EcoRI site on the pcDNA3.1 plasmid, which was controlled by the CMV promoter, to form the recombinant expressing vector pcDNA3.1-GM-CSF. Results: The PCR amplification was identified and the sequence was analyzed, the results showed that hGM-CSF was properly inserted into the vector and the sequence was correct.