Preclinical evaluation of herpes simplex virus armed with granulocyte-macrophage colony-stimulating factor in pancreatic carcinoma

AIM: To investigate the therapeutic efficacy and mechanisms of action of oncolytic-herpes-simplex-virus encoding granulocyte-macrophage colony-stimulating factor(HSVGM-CSF) in pancreatic carcinoma.METHODS: Tumor blocks were homogenized in a sterile grinder in saline.The homogenate was injected into the right armpit of each mouse.After vaccination,the mice were randomly assigned into four groups: a control group,a high dose HSVGM-CSFgroup [1 × 107plaque forming units(pfu)/tumor],a medium dose HSVGM-CSF group(5 × 106pfu/tumor) and a low dose HSVGM-CSF group(5 × 105pfu/tumor).After initiation of drug administration,body weights and tumor diameters were measured every 3 d.Fifteen days later,after decapitation of the animal by cervical dislocation,each tumor was isolated,weighed and stored in 10% formaldehyde solution.The drug effectiveness was evaluated according to the weight,volume and relative volume change of each tumor.Furthermore,GM-CSF protein levels in serum were assayed by enzyme-linked immunosorbent assays at 1,2,3 and 4 d after injection of HSVGM-CSF.RESULTS: Injection of the recombinant mouse HSV encoding GM-CSF resulted in a significant reduction in tumor growth compared to the control group,and dosedependent effects were observed: the relative tumor proliferation rates of the low dose,medium dose and high dose groups on 15 d after injection were 45.5%,55.2% and 65.5%,respectively.The inhibition rates of the tumor weights of the low,middle,and high dose groups were 41.4%,46.7% and 50.5%,respectively.Furthermore,the production of GM-CSF was significantly increased in the mice infected with HSVGM-CSF.The increase in the GM-CSF level was more pronounced in the high dose group compared to the other two dose groups.CONCLUSION: Our study provides the first evidence that HSVGM-CSFcould inhibit the growth of pancreatic cancer.The enhanced GM-CSF expression might be responsible for the phenomenon.

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.

Granulocyte-macrophage colony-stimulating factor-transfected bone marrow stromal cells for the treatment of ischemic stroke

Adult, male, Sprague-Dawley rats were injected with granulocyte-macrophage colony-stimulating factor-transfected bone marrow stromal cells （GM-CSF-BMSCs） into the ischemic boundary zone at 24 hours after onset of middle cerebral artery occlusion. Results showed reduced infarct volume, decreased number of apoptotic cells, improved neurological functions, increased angiogenic factor expression, and increased vascular density in the ischemic boundary zone in rats that underwent GM-CSF-BMSCs transplantation compared with the BMSCs group. Experimental findings suggested that GM-CSF-BMSCs could serve as a potential therapeutic strategy for ischemic stroke and are superior to BMSCs alone.

EFFECTS OF GRANULOCYTE-MACROPHAGE COLONY STIMULATING FACTOR GENE ENCODED VACCINIA VIRUS VECTOR ON MURINE PULMONARY METASTATIC MELANOMA

A recombinant vaccinia virus expressing murine granulocyte-macrophage colony-stimulating factor (VVGM-CSF) was tested for its antitumor activity.Murine pulmonary metastasis was established by injecting 20×10~5 B16F10 melanoma cells into the tail vein of C57BL/6 mice. Three days after B16F10 inoculation,WGM-CSF or VVTK, a thymidine kinase gene deficient control vaccinia virus, were injected intraperitoneally twice weekly for 2 weeks. Two weeks later, the mice were sacrificed and pulmonary metastasis fool counted.The results demonstrated that VVGM-CSF treatment significantly decreased the number of pulmonary metastasis and prolonged the survival time of tumorbearing mice. Cytotoxic and phagocytic activities of the peritoncal macrophages were found to be markedly elevated in mice treated with WGM-CSF. Nitric oxide released from the macrophages was also found to be increased. These data, together with our other results,strongly demonstrated that continuous secretion of GMCSF and activation of macrophages might pal-tially explain the therapeutic effects of VVGM-CSF on murine pulmonary metastasis.

Pharmacokinetics of recombinant human granulocyte macrophage colony-stimulating factor in Macaca mulata

目的：研究重组人粒细胞巨噬细胞集落刺激因子（ｒｈＧＭＣＳＦ）在恒河猴体内的药物动力学．方法：用酶连接免疫吸附测定法检测血浆中ｒｈＧＭＣＳＦ的含量．结果：ｉｖｒｈＧＭＣＳＦ后血药浓度时间曲线符合三房室模型．第１，２和３相的Ｔ１２分别为００５－００７ｈ，０１４－０５８ｈ和１４－４１ｈ．ＡＵＣ随剂量成比例增加．ｉｖ高剂量和低剂量的Ｃｌ和Ｋ１０都相似．ｓｃｒｈＧＭＣＳＦ后血药浓度的峰值为０９３±０１６μｇ·Ｌ－１，达峰时间为２６５±０１４ｈ，生物利用度为０６１．结论：恒河猴ｒｈＧＭＣＳＦ药物动力学数据为临床试验提供有用参考．

A pilot study on the combined therapy of granulocyte-macrophage colony-stimulating factor and hepatitis B vaccine on chronic hepatitis B virus carrier children

Objective To observe the efficacy of treating intrauterine infected chronic hepatitis B virus (HBV) carrier children with a combination of granulocyte macrophage colony stimulating factor (GM CSF) or hepatitis B immunoglobulin (HBIG) plus recombinant hepatitis B vaccine (rHBvac) Methods A total of 27 chronic HBV infected children, who were born to HBV carrier mothers and received hepatitis B immunoprophylaxis at birth, were randomized into 2 groups: one receiving a combined therapy of 50 μg of GM CSF plus 10 μg of rHBvac injected intramuscularly at the same location (GM CSF group, 14 children) or 200 IU HBIG and 10 μg rHBvac in different muscles (HBIG group, 13 children) on a monthly four dose schedule HBV DNA quantification and other HBV serological markers were tested before and after the four dose therapy Results Twelve children in each group completed the study Of them, 3 children in the GM CSF group and 4 in the HBIG group had elevated serum alanine transaminase (ALT) before the trial, and then 2 in each group became ALT normal after the treatment Before the therapy, hepatitis B e antigen (HBeAg) positivity was found in nine children in the GM CSF group and 10 in the HBIG group One from each group had an HBeAg/anti HBe seroconversion after the treatment The quantity of HBV DNA was significantly lower after the treatment ( P =0 023) in GM CSF group, but was not significantly reduced in HBIG group No subjects were found to be negative for hepatitis B surface antigen (HBsAg) after the treatment, and no serious adverse events occurred in either group Conclusion Combined GM CSF and rHBvac therapy inhibit HBV replication in carrier children who were not protected after treatment with immunoprophylaxis

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。

血管瘤患儿血清粒细胞-巨噬细胞集落刺激因子及转录激活因子3水平与瘤体增生的相关性

目的探索血管瘤患儿血清粒细胞-巨噬细胞集落刺激因子(GM-CSF)及转录激活因子3(STAT3)的表达水平并分析其与瘤体增生的相关性。方法选取2020年1月—2023年1月收治的152例血管瘤增生患儿作为研究对象,分为增生期组(87例)与退化期组(65例),另选择同期确诊为血管畸形的53例患儿作为血管畸形组。对血清GM-CSF、STAT3及碱性成纤维细胞生长因子(bFGF)、血管内皮生长因子(VEGF)、缺氧诱导因子-1α(HIF-1α)、血管生成素-1(Ang-1)的表达水平进行检测;Pearson法相关性分析血清GM-CSF、STAT3与bFGF、VEGF、HIF-1α、Ang-1的相关性;受试者工作特征(ROC)曲线分析血清GM-CSF、STAT3对血管瘤瘤体增生的预测价值;多因素Logistic回归分析血管瘤瘤体增生的影响因素。结果增生期组患儿血清bFGF、VEGF、HIF-1α、GM-CSF、STAT3表达水平高于血管畸形组及退化期组患儿(P<0.05),但血清Ang-1表达水平低于血管畸形组及退化期组患儿(P<0.05);Pearson法相关性分析显示,增生期组患儿血清GM-CSF及STAT3水平均与bFGF、VEGF、HIF-1α水平正相关,与Ang-1水平负相关;ROC曲线下面积显示,GM-CSF和STAT3单独预测血管瘤瘤体增生风险的AUC分别为0.847,0.822,联合预测的AUC为0.918,联合预测的效能显著大于GM-CSF单独诊断的AUC(Z=2.459,P=0.014),STAT3单独诊断的AUC(Z=3.371,P=0.001)。多因素Logistic回归分析显示GM-CSF、STAT3是血管瘤瘤体增生的影响因素(P<0.05)。结论血管瘤瘤体增生患儿血清GM-CSF及STAT3表达水平升高,是瘤体增生的影响因素,联合检测能较好的预测血管瘤瘤体增生风险。

Granulocyte-macrophage colony-stimulating factor and interleukin 4 induce the malignant transformation of the bone marrow- derived human adult mesenchymal stem cells

Background The purpose of the study was to examine the effects of granulocyte-macrophage colony-stimulating factor （GM-CSF） and interleukin 4 （IL-4） on the bone-marrow-derived human adult mesenchymal stem cells （hMSCs）. Methods The hMSCs were isolated and cultured with GM-CSF and IL-4 for a period of one month. A single colony of transformed cells was then isoloated and their phenotype was characterized by morphology, surface marker expression, and in vivo tumorigenesis.Results After one month culture, the transformed mesenchymal cells exhibited the morphology and phenotype similar to those of tumor cells, and also caused multiple fast growing lung deposits when it was injected into immunodeficient mice.Conclusion Cytokines-driven malignant transformation of hMSCs may be a useful model for studying signaling pathways initiating malignant transformation of hMSC.

Effects of granulocyte-macrophage colony stimulating factor on the repair of vessel intima damaged by balloon

The dysfunction of vascular endothelial cells plays a key role in startingand facilitating restenosis. The acceleration of intima repair and the recovery of endothelialfunction would reduce the restenosis rate. This study was undertaken to assess the effect ofgranulocyte-macrophage colony stimulating factor ( GM-CSF) on the repair of damaged iliac arteries.Twenty-four male New Zealand white rabbits undergoing primary iliac artery deendothelialization wererandomly divided into two groups ( GM-CSF group and control group) . The GM-CSF group received asubcutaneous injection of GM-CSF (10 μg ? kg^(-1) ? d^(-1) ) , and the control groupwas given a subcutaneous injection of equivalent saline. The iliac arteries of all animals weredamaged by balloon after 7 days. The levels of nitric oxide ( NO) were detected before, 1 week, 2weeks and 4 weeks after angioplasty. The repair and hyperplasia of the intima were observedmicroscopically and the indices of stenosis were evaluated by computerized planimetry after 4 weeksof angioplasty. The NO levels of the GM-CSF group were higher than those of the control group 2weeks and 4 weeks after angioplasty [91.92 +-11.57) μmol/L vs. (81. 67 +- 12. 18) μmol/L; (97. 67+- 10. 13 ) ( μmol/L vs. (83. 16 +-12. 64) μmol/L]. Four weeks after balloon damage,histological examination showed that neointima formation, vascular smooth muscle cells and fibroustissue of the GM-CSF group were less than those of the control group. The endothelium of the GM-CSFgroup was more integrated, and stenosis of lumen was slighter than that of the control group.Morphometry showed the lumen area of the GM-CSF group was larger than that of the control group[(1.27 +-0. 31) mm^2 vs. (0. 92 +- 0. 24) mm^2 ] , the neointimal area and percent of intimahyperplasia were significantly smaller than those of the control group [ (0. 85 +-0. 34) mm vs. (1.18 +-0. 38) mm^2; (40 +- 7)% vs. (55 +- 6)%]. GM-CSF could facilitate the repair of the intima,reduce neointima formation, better the function of the endothelium, and decrease the rate ofrestenosis.

金刚藤胶囊联合桂枝茯苓胶囊治疗慢性盆腔炎的疗效及对血清GM-CSF、MMP-2水平的影响

【目的】分析金刚藤胶囊联合桂枝茯苓胶囊治疗湿热瘀结型慢性盆腔炎的疗效及对血清粒细胞-巨噬细胞集落刺激因子(GM-CSF)、基质金属蛋白酶2(MMP-2)水平的影响。【方法】将90例湿热瘀结型慢性盆腔炎患者随机分为联合组和单药组,每组各45例。单药组患者给予桂枝茯苓胶囊治疗,联合组患者在单药组的基础上联合金刚藤胶囊治疗,疗程为2周并随访1个月。观察2组患者治疗前后中医证候积分和血清GM-CSF、MMP-2水平的变化情况,比较2组患者的临床疗效、症状缓解时间、疾病复发和不良反应发生情况。【结果】(1)疗效方面,治疗2周后,联合组的总有效率为93.33%(42/45),单药组为66.67%(30/45),组间比较,联合组的疗效明显优于单药组(P<0.01)。(2)中医证候积分方面,治疗后,2组患者的下腹疼痛、腰骶疼痛、带下量多、月经量多、经行腹痛、神疲乏力等各项中医证候积分均较治疗前明显降低(P<0.05),且联合组对各项中医证候积分的降低幅度均明显优于单药组(P<0.05)。(3)症状缓解时间方面,联合组患者治疗后的白带恢复正常时间、下腹坠胀和腹痛缓解时间均较单药组明显缩短(P<0.01)。(4)血清学指标方面,治疗后,2组患者的血清GM-CSF、MMP-2水平均较治疗前明显降低(P<0.05),且联合组对血清GM-CSF、MMP-2水平的降低幅度均明显优于单药组(P<0.05或P<0.01)。(5)疾病复发和不良反应方面,联合组的复发率和不良反应发生率分别为11.11%(5/45)、13.33%(6/45),均明显低于单药组的35.56%(16/45),差异均有统计学意义(P<0.05或P<0.01)。【结论】金刚藤胶囊联合桂枝茯苓胶囊治疗,可以显著提高湿热瘀结型慢性盆腔炎的临床疗效,有效缩短症状缓解时间,降低血清GM-CSF、MMP-2水平。

干扰素调节因子5、粒细胞-巨噬细胞集落刺激因子在细菌性肺炎病儿血清中的表达及预后价值

目的探讨干扰素调节因子5(interferon regulatory factor 5,IRF5)、粒细胞-巨噬细胞集落刺激因子(granulocyte-macro-phage colony-stimulating factor,GM-CSF)在细菌性肺炎病儿血清中的表达变化及疾病预后价值。方法选取2019年4月至2021年5月中国人民解放军联勤保障部队第九二八医院收治的细菌感染性肺炎病儿96例作为细菌组,同期非细菌感染性肺炎病儿88例作为非细菌组,另外选取健康体检儿童96例为对照组,收集检测三组病儿基本临床资料。检测血清中IRF5、GM-CSF及其组间差异;采用Pearson法分析细菌性肺炎病儿血清IRF5、GM-CSF水平与病情指标的相关性;应用受试者操作特征曲线(ROC曲线)分析IRF5、GM-CSF及二者联合预测细菌性肺炎预后的价值。结果细菌组血清IRF5水平[(38.85±13.86)ng/L]显著高于非细菌组[(11.15±4.37)ng/L]和对照组[(10.76±1.55)ng/L],且重症组高于轻症组(P<0.05);细菌组血清GM-CSF水平[(54.73±16.56)ng/L]显著低于非细菌组[(246.73±28.94)ng/L]和对照组[(250.64±55.67)ng/L],且重症组低于轻症组(P<0.05)。重症组气促、吐沫、三凹征、肺部明显湿啰音比例、C反应蛋白(C-reactive protein,CRP)、白细胞水平显著高于轻症组、非细菌组(P<0.05),轻症组、非细菌组CRP水平显著高于对照组(P<0.05),非细菌组CRP水平显著高于对照组(P<0.05)。血清IRF5水平与CRP、白细胞、临床肺部感染评分(clinical pulmonary infection score,CPIS)均呈正相关(r=0.34、0.36、0.41,P<0.05),血清GM-CSF水平与CRP、白细胞、CPIS均呈负相关(r=-0.40、-0.32、-0.45,P<0.05)。预后不良组血清IRF5水平高于预后良好组,血清GM-CSF水平低于预后良好组(P<0.05)。ROC曲线显示,IRF5、GM-CSF对预后预测的AUC分别为0.90、0.87,二者联合对预后预测的AUC为0.96,明显高于二者单独诊断,(Z联合vs.IRF5=2.86,P=0.004;Z联合vs.GM-CSF=2.24,P=0.025),其灵敏度、特异度分别为90.32%、90.77%。结论细菌性肺炎病儿血清IRF5水平升高,GM-CSF水平降低,二者在评估病情进展及疾病预后预测方面具有较高的价值。

进行气管插管的颅脑外伤患者血清GM-CSF和HMGB1水平及其与并发肺部感染的关系

目的探讨血清粒-巨噬细胞集落刺激因子(GM-CSF)、高迁移率族蛋白B1(HMGB1)与进行气管插管的颅脑外伤患者并发肺部感染的关系。方法选取2020年12月至2022年12月华北医疗健康集团峰峰总医院收治的156例进行气管插管的颅脑外伤患者作为颅脑外伤组,根据是否出现肺部感染将进行气管插管的颅脑外伤患者分为感染组和未感染组,另根据临床肺部感染评分(CPIS)将进行气管插管的颅脑外伤并发肺部感染患者分为重度感染组和轻度感染组,并选取同期在华北医疗健康集团峰峰总医院体检的体检健康者60例作为对照组。采用酶联免疫吸附试验(ELISA)测定所有研究对象血清GM-CSF、HMGB1水平;分析进行气管插管的颅脑外伤并发肺部感染患者血清GM-CSF、HMGB1水平与CPIS的关系;采用多因素Logistic回归分析进行气管插管的颅脑外伤患者并发肺部感染的影响因素;采用受试者工作特征(ROC)曲线分析血清GM-CSF、HMGB1对进行气管插管的颅脑外伤患者并发肺部感染的预测价值。结果与对照组相比,颅脑外伤组患者血清GM-CSF、HMGB1水平均明显升高(P<0.05)。感染组患者血清GM-CSF、HMGB1水平显著高于未感染组(P<0.05)。重度感染组21例,轻度感染组45例。与轻度感染组患者相比,重度感染组患者血清GM-CSF、HMGB1水平及CPIS均显著升高(P<0.05)。相关分析结果显示,感染组患者血清GM-CSF、HMGB1水平与CPIS均呈正相关(r=0.408、0.435,P<0.05)。多因素Logistic回归分析结果显示,年龄≥60岁、入院时GCS评分≥6分、合并糖尿病、GM-CSF水平升高、HMGB1水平升高均是进行气管插管的颅脑外伤患者并发肺部感染的危险因素(P<0.05)。ROC曲线分析结果显示,GM-CSF、HMGB1联合预测进行气管插管的颅脑外伤患者并发肺部感染的曲线下面积(AUC)为0.912(95%CI:0.860~0.963),明显大于GM-CSF、HMGB1单项预测的AUC[0.826(95%CI:0.756~0.895)、0.819(95%CI:0.743~0.896)],差异均有统计学意义(Z=1.972,P=0.049;Z=1.984,P=0.047)。结论血清GM-CSF、HMGB1水平在进行气管插管的颅脑外伤并发肺部感染的患者中升高,是进行气管插管的颅脑外伤患者发生肺部感染的危险因素,2项指标联合检测对进行气管插管的颅脑外伤患者并发肺部感染具有良好的预测价值。

血清GM-CSF、IGF-1联合检测在妊娠期糖尿病患者不良妊娠结局诊断中的价值

目的:分析血清粒细胞-巨噬细胞集落刺激因子(GM-CSF)、胰岛素样生长因子-1(IGF-1)联合检测在妊娠期糖尿病(GDM)患者不良妊娠结局诊断中的价值。方法:选取2021年6月至2023年3月该院收治的83例GDM患者设为研究组,根据妊娠结局将其分为不良结局患者(n=24)和良好结局患者(n=59),另选取同期160名健康孕妇作为对照组,比较两组及不同妊娠结局GDM患者血清GM-CSF、IGF-1水平,绘制受试者工作特征(ROC)曲线,分析血清GM-CSF、IGF-1单项及联合检测在GDM患者不良妊娠结局诊断中的价值。结果:研究组血清IGF-1水平低于对照组,GM-CSF水平高于对照组,差异均有统计学意义(P<0.05);不良结局患者IGF-1水平低于良好结局患者,GM-CSF水平高于良好结局患者,差异均有统计学意义(P<0.05);ROC曲线分析结果显示,血清IGF-1、GM-CSF单项及联合检测诊断GDM患者不良妊娠结局的曲线下面积分别为0.821、0.822、0.912,均具有一定诊断价值,且联合检测诊断价值最高。结论:血清GM-CSF、IGF-1联合检测在GDM患者不良娠结局诊断中的价值高于二者单项检测。

魏氏验方联合双氯芬酸钠治疗冻结期肩周炎的效果及免疫-炎症机制

目的探讨魏氏验方联合双氯芬酸钠治疗冻结期肩周炎的效果及免疫-炎症机制。方法选取2020年1月—2022年6月收治的120例冻结期肩周炎为研究对象,随机数字表法分为对照组和研究组,每组60例。对照组采取双氯芬酸钠治疗,研究组采取魏氏验方(伸筋活血汤+痹通洗方)联合双氯芬酸钠治疗,连续治疗3个月。观察对比2组中医疗效、中医证候积分、免疫-炎症相关指标[粒细胞-巨噬细胞集落刺激因子(GM-CSF)、白细胞介素-10(IL-10)、IL-6、肿瘤坏死因子-α(TNF-α)]、疼痛因子[前列腺素E_(2)(PGE_(2))、β内啡肽(β-EP)]、肩关节功能相关指标及不良反应发生情况。结果治疗3个月后,研究组治疗总有效率90.00%(54/60)高于对照组75.00%(45/60)(P<0.05)。治疗1、3个月后,2组关节疼痛、肿胀、沉重、畏寒积分以及血清GM-CSF、IL-10、IL-6、TNF-α、PGE_(2)水平均低于治疗前,且研究组低于对照组(P<0.05)。治疗1、3个月后,2组肩关节被动及主动状态前屈和外展活动度、MMT评分、Constant-Murley评分、血清β-EP水平均高于治疗前,且研究组高于对照组(P<0.05)。2组不良反应发生率比较差异无统计学意义(P>0.05)。结论魏氏验方联合双氯芬酸钠能减轻冻结期肩周炎患者炎症反应,缓解疼痛,改善临床症状及肩关节功能,且效果确切、安全性高。

链霉亲和素-人粒细胞巨噬细胞集落刺激因子融合蛋白的制备与活性鉴定

目的:制备链霉亲和素(SA)连接的人粒细胞巨噬细胞集落刺激因子(hGM-CSF)融合蛋白(SA-hGM-CSF),并对其生物学活性进行鉴定。方法:在NCBI数据库中查找SA和hGM-CSF序列,将序列合成后连接入Pet28表达载体中,构建SA-hGM-CSF-Pet28重组表达载体,在BL21(DE3)表达感受态中利用IPTG诱导剂诱导表达,经His镍柱纯化,尿素梯度复性,聚丙烯酰胺凝胶电泳法和蛋白质印迹法(WB)鉴定融合蛋白,通过TF-1细胞检测融合蛋白的生物学活性。结果:SA-hGM-CSF融合蛋白可在BL21(DE3)中高效诱导表达,经His镍柱纯化后该融合蛋白纯度较高。经尿素梯度复性后,细胞学实验验证该融合蛋白具有生物学活性。结论:成功表达具有生物活性的融合蛋白SA-hGM-CSF,该结果为SA-hGM-CSF表面锚定修饰的肿瘤细胞疫苗的研究提供了实验基础。

立体定向体部放疗联合粒细胞-巨噬细胞集落刺激因子治疗原发性肝癌患者的效果

目的探讨立体定向体部放疗(SBRT)联合粒细胞-巨噬细胞集落刺激因子(GM-CSF)治疗原发性肝癌患者的效果。方法纳入接受SBRT联合GM-CSF治疗的31例原发性肝癌患者。观察治疗后患者的近期疗效(疾病控制率、客观缓解率)和远期疗效(无病生存期、总生存期),分析治疗前后患者的免疫指标(CD4^(+)T淋巴细胞水平、CD8^(+)T淋巴细胞水平及CD4^(+)/CD8^(+)值)、肝功能指标(ALT、AST、总胆红素、白蛋白水平)、肝癌指标(甲胎蛋白、癌胚抗原水平)、生活质量[Karnofsky功能状态(KPS)评分]的变化。结果治疗后1个月、3个月、6个月和12个月的客观缓解率分别为77.42%、90.00%、58.62%和40.74%,疾病控制率分别96.77%、93.33%、75.86%和70.37%。31例患者的中位无病生存期为9.1个月,中位总生存期为21.0个月。不同临床特征患者的中位无病生存期和中位总生存期差异均无统计学意义(均P>0.05)。与治疗前1周比较,治疗后1周患者的CD4^(+)T淋巴细胞水平、CD8^(+)T淋巴细胞水平、CD4^(+)/CD8^(+)值、白蛋白水平下降,ALT、AST水平升高(均P<0.05);与治疗后1周比较,治疗后1个月、3个月患者的CD4^(+)T淋巴细胞水平、CD8^(+)T淋巴细胞水平、CD4^(+)/CD8^(+)值升高(均P<0.05),而ALT、AST、白蛋白水平恢复至治疗前水平。治疗后1周、1个月、3个月患者的血清癌胚抗原、甲胎蛋白水平较治疗前1周有所下降,治疗后患者的KPS评分呈先降低后升高的趋势(均P<0.05)。结论SBRT联合GM-CSF可改善原发性肝癌患者的生活质量,维持患者肝功能的稳定,增强患者的免疫功能,提高患者远期临床疗效,可作为失去手术指征的原发性肝癌患者的治疗方案。

Regulatory effects of lipopolysaccharide in murine macrophage proliferation

AIMS To study the regulatory effects of bacterial lipopolysaccharide (LPS) in murine macrophage proliferation . METHODS Using murine peritoneal exudate macrophage (PEM) and macrophage cell line J 774 A.1 as targets, LPS effects on M CSF and granulocyte macrophage colony stimulating factor (GM CSF) stimulated macrophage colony forming cells (CFU M) were detected. 125 I GM CSF receptor binding assay was used to examine LPS regulation on GM CSF receptor expression. RT PCR was employed to test TGF β 1 inhibition on IFN γ mRNA expression on macrophage induced by LPS. RESULTS Without direct effect on macrophage proliferation, LPS could inhibit the macrophage proliferation stimulated by GM CSF. However, with the concomitant existence of GM CSF and TGF β 1, the LPS inhibitory effect was eliminated. RT PCR analysis indicated that the strongest macrophage growth inhibitory factor IFN γ mRNA expression in macrophage induced by LPS was remarkably suppressed by TGF β 1, 125 I GM CSF receptor binding assay showed that LPS could enhance GM CSF receptor expression likewise as TGF β 1 . CONCLUSIONS LPS is involved in the network of macrophage proliferative regulation by multiple cytokines, displaying inhibitory and stimulatory effects based on the coexisting cytokines.

PRP凝胶与rhGM-CSF凝胶治疗慢性难愈性创面的效果及对创面修复因子的影响

目的比较自体富血小板血浆(PRP)凝胶与重组人粒细胞巨噬细胞集落刺激因子(rhGM-CSF)凝胶治疗慢性难愈性创面的效果及对创面修复因子的影响。方法将80例慢性难愈性创面患者分为PRP组和rhGM-CSF组,各40例。在外科常规治疗基础上,PRP组采用PRP凝胶治疗,rhGM-CSF组采用rhGM-CSF凝胶治疗。比较两组患者创面完全愈合时间,治疗2周后创面愈合率、感染控制水平,治疗前后创面修复因子[血管内皮生长因子(VEGF)、血小板衍生生长因子(PDGF)、碱性成纤维细胞生长因子(bFGF)、表皮生长因子(EGF)]表达量、疼痛视觉模拟量表(VAS)评分。记录两组患者治疗过程中不良反应的发生情况。结果PRP组创面完全愈合时间短于rhGM-CSF组(P<0.05)。治疗2周后,PRP组患者创面愈合率高于rhGM-CSF组;两组患者疼痛VAS评分、白细胞计数及血清C反应蛋白水平均低于治疗前,创面VEGF、PDGF、bFGF、EGF表达量均较治疗前升高,且PRP组患者上述指标均优于rhGM-CSF组(均P<0.05)。两组患者治疗前后的创面细菌培养阳性情况差异无统计学意义(P>0.05)。治疗过程中PRP组无不良反应发生,rhGM-CSF组共出现3例不良反应,两组不良反应总发生率差异无统计学意义(P>0.05)。结论PRP凝胶与rhGM-CSF凝胶对慢性难愈性创面患者的创面愈合情况、疼痛、感染水平等均具有改善作用,而PRP凝胶上述效果优于rhGM-CSF凝胶。

rhGM-CSF凝胶联合磺胺嘧啶银对面部深Ⅱ度烧伤创面愈合质量的影响

目的:探讨重组人粒细胞巨噬细胞集落刺激因子(Recombinant human granulocyte macrophage colony stimulating factor,rhGM-CSF)联合磺胺嘧啶银治疗面部深Ⅱ度烧伤的疗效。方法:选择2016年6月-2021年11月笔者医院收治的96例面部深Ⅱ度烧伤患者,采用随机数表法分为观察组和对照组各48例,对照组给予磺胺嘧啶银霜治疗,观察组给予磺胺嘧啶银霜+rhGM-CSF凝胶治疗,比较两组创面愈合相关指标及美学效果。结果:观察组创面结痂时间、痂皮基本溶解时间及创面完全愈合时间均短于对照组(P<0.05),治疗后各时间点疼痛评分均低于对照组(P<0.05);治疗后,两组创面细菌培养阳性率均降低,但组间比较差异无统计学意义(P>0.05);治疗后1、2周,两组血清血管内皮生长因子(Vascular endothelial growth factor,VEGF)、表皮生长因子(Epidermal growth factor,EGF)、转化生长因子β1(Transforming growth factor-β1,TGF-β1)水平均高于治疗前,且观察组高于对照组(P<0.05);治疗3周后,观察组创面美学效果各评分及总分均明显高于对照组(P<0.05);两组并发症发生率比较差异无统计学意义(P>0.05)。结论:对面部深Ⅱ度烧伤患者采用rhGM-CSF凝胶与磺胺嘧啶银联合治疗,可有效促进患者创面愈合、降低疼痛程度,提高血管生长因子水平,美学效果好,值得在临床推广应用。