The in vitro effects of cytokines on expansion and migration of megakaryocyte progenitors

Increasing the number of megakaryocyte progenitors in stem cell transplants by ex vivo expansion culture may be an approach to accelerate platelet recovery in patients undergoing high-dose chemotherapy. We evaluated the effect of three different cytokine combinations on expansion, with special emphasis on the type of colony formation and migration of megakaryocytic cells. The number of clonogenic megakaryocyte progenitors (colony-forming units-megakaryocyte; CFU-Mk) with high- (> 20 cells/colony) and low-proliferative capacity (5-20 cells/colony) and the number of megakaryocytic (CD61+) cells were significantly increased by including interleukin 3 (IL-3) or IL-3 + IL-6 + IL-11 + Flt3-ligand to cultures containing megakaryocyte growth and development factor (MGDF) plus stem cell factor (SCF). No difference in the maturation of megakaryocytes from all three cytokine combinations to platelets were observed, as demonstrated by electron microscopy. In chemotaxis experiments, the migration towards stromal cell-derived factor 1 (SDF-1) was shown to be reduced for CD61+ cells and megakaryocyte progenitors cultured in other cytokines besides MGDF + SCF. The reduced migration was related to a lower expression of CXCR4, the receptor for SDF-1, on megakaryocytes from the proliferating cultures. These in vitro results demonstrate that expansion in IL-3 and other cytokines besides MGDF + SCF significantly impair the capacity of megakaryocytic cells to migrate.     

Treatment of enteritis in chronic granulomatous disease with granulocyte colony stimulating factor

Background—In several diseases there is a relationbetween deficiency of neutrophil granulocytes and granulomatouslesions. Recently, in glycogen storage disease type Ib, this relationhas been supported by the beneficial effect of treatment of enteritis with granulocyte-macrophage colony stimulating factor.
Aim—To investigate whether chronic granulomatousdisease could be treated according to the same principle.
Patients and methods—Inflammatory lesions weremonitored in two brothers with chronic granulomatous diseasedemonstrated by very low superoxide production in neutrophilgranulocytes. The two patients were treated with recombinant humangranulocyte colony stimulating factor on three occasions when thedisease was active.
Results—In one patient, remission of an inflamedstenosis of the colon sigmoideum was shown by granulocytescintigraphy after one month of treatment with granulocyte colonystimulating factor. In the other patient, remission of colon diseaseand later of a non-malignant tumour in the right lung hilum was shownby colonoscopy and computed tomography scans respectively.
Conclusion—Remission of inflammatory lesions intwo brothers with chronic granulomatous disease was induced bygranulocyte colony stimulating factor on three occasions. The mechanismfor this effect is not known. The result is similar to the response found in patients with leucocyte deficiency due to glycogen storage disease type Ib.

Keywords:chronic granulomatous disease; enteritis; granulocyte colony stimulating factor

     

武警官兵《心理辅导需要问卷》的编制与信效度检验

目的编制适用于武警官兵的心理辅导需要问卷。方法通过开放式问卷和访谈收集初始问卷题目,采用探索性和验证性因素分析对问卷进行检验。结果武警官兵心理辅导需要包含4个维度共25个条目,维度的单因素模型拟合指数良好,内部一致性系数为0．92,4个因子的内部一致性系数在0．71～0．92。结论问卷具有良好的信度和效度,可以作为武警官兵心理辅导需要分析的工具。     

数字化成形三维钛网在颅骨缺损修补手术中的应用

目的比较预塑球形钛网和数字化成形三维钛网修补颅骨缺损的临床效果,探讨数字化成形三维钛网在颅骨修补手术中的操作要点及应用价值。方法回顾性分析我院2001年8月至2009年1月155例采用预塑球形钛网行颅骨修补术的患者临床资料,与2009年2月至2013年3月83例采用数字化成形三维钛网行颅骨修补术的患者临床资料进行比较。结果手术时间:预塑钛网组(125±8)min,三维钛网组(98±6)min,预塑钛网组平均手术时间明显长于三维钛网组(P0.05);使用钛钉数量:预塑钛网组(11.8±1.5)个,三维钛网组(8.3±1.2)个,预塑钛网组平均使用钛钉数量明显多于三维钛网组(P0.05);外观满意度:预塑钛网组(7.30±0.21)分,三维钛网组(10.00±0.00)分,预塑钛网组外观满意度明显低于三维钛网组(P0.05);术后并发症:预塑钛网组共19例,发生率12.26%,三维钛网组共3例,发生率3.61%,预塑钛网组术后并发症明显高于三维钛网组(P0.05)。结论采用数字化成形三维钛网修补颅骨缺损具有手术时间短,使用钛钉数量少,外观满意度高,术后并发症少优点,是目前颅骨修补手术的首选材料,应该在临床推广应用。     

醋酸亮丙瑞林原位注射制剂的制备及其体内外释放性能的研究

目的：合成mPEG/PLA比例为10/90,5/95,2/98的3种聚合物并对其进行结构表征。用合成的mPEG-PLA为载体材料和有机溶剂N-甲基比咯烷酮（NMP）制备醋酸亮丙瑞林（LA）的原位注射剂并对其物理化学性质和体内外释放试验进行研究。方法：采用mPEG2000和外消旋-丙交酯（DL-LA）,以开环聚合法合成比例为10/90,5/95,2/98的聚合物mPEG-PLA,并通过1H-NMR对产物进行结构确证。然后将mPEG-PLA和LA置于两个注射器中,制成A/B系统,在使用前将药物加入聚合物溶液中并混合均匀,制备醋酸亮丙瑞林的原位注射剂。利用显微镜和SEM分析制剂结构。其次进行药物的体外释放试验,对释放曲线进行模型拟合,最后以大鼠为动物模型进行试验,考察药物在SD大鼠体内的释放行为,并考察体内外相关性。结果：成功合成且验证了不同比例的mPEG/PLA聚合物,并以此为载体材料成功制备出LA原位注射制剂。体外释放试验表明,药物以接近零级的方式释放,释放速率和聚合物mPEG/PLA比例,载药量和加入有机溶剂的量有关,且聚合物中PLA比例越高释放越慢,载药量增加则释放速率增加;选用黏度大的溶剂则药物释放减慢,溶剂的用量增加释放速率减小;大鼠体内实验也表明该制剂在体内能够达到缓释的效果,药物在前10 d内释放较快,30 d释放药物能达到90%。结论：醋酸亮丙瑞林原位注射制剂物理化学性质稳定,体内外实验均证明该制剂具有良好的释放性能。     

Designing the epitope flanking regions for optimal generation of CTL epitopes

The flanking amino acids that surround epitopes are critical for effective antigen processing and maintenance of epitope integrity. In the present study, the frequency and characteristics of each amino acid that flanked the peptides generated from the proteasomal degradation of three different subtypes of HIV-1 Gag-p24 were determined. Synthetic flanking regions were designed based on the highest and the lowest frequencies of amino acid with the ideal characteristics at positions upstream and downstream of the proteasomal cleavage site. Peptides were synthesized that contained known CD8+ CTL-epitopes from HIV-1 Gag, CMV pp65, and vaccinia proteins HRP-2, and C16, flanked by amino acid sequences specifically designed to either generate or inhibit the known CD8+ CTL-epitopes. As predicted, the known CD8+ CTL-epitopes were effectively generated from the peptides with synthetic flanking regions specifically designed to promote epitope generation in a proteasome-dependent manner. The majority of the proteasome-generated epitopes were cleaved immediately after the C-terminal amino acid of the specific CTL-epitope. The synthetic peptide sequences containing known CD8+ CTL-epitopes with the flanking regions that promote epitope generation were effectively processed and presented to epitope specific CD8+ T-cells resulting in the production of IFN-γ. These results highlight the importance of flanking regions in promoting efficient antigen processing and presentation. This concept can have important implications in vaccine design and development strategies.     

脑胶质瘤耐放射细胞亚株的建立方法

目的探索诱导并建立脑胶质瘤耐放射细胞亚株的体外实验方法,为进一步研究胶质瘤放疗抵抗发生的分子生物学机制提供基础。方法以脑胶质瘤U251细胞株为实验对象,通过体外细胞培养,应用60Co射线对脑胶质瘤U251细胞株进行小剂量重复的诱导照射,得到U251耐放射细胞亚株RR-U251。观察RRU251细胞形态的变化;MTT检测其细胞活力,流式细胞仪(FCM)检测其细胞凋亡;集落形成实验检测细胞克隆团形成。结果放射治疗后,RR-U251与U251细胞的相比:细胞增殖活力增强11.5%,迁移能力增强50%,侵袭能力增强87.5%,凋亡率下降69.6%,细胞集落形成率升高88.5%;RR-U251放射敏感性明显下降。结论小剂量重复照射诱导法能较好地建立胶质瘤耐放射细胞亚株RR-U251。     

神阙灸防治ICU肠内营养并发腹泻的临床疗效观察

目的：观察艾灸神阙穴防治ICU肠内营养并发腹泻的临床疗效。方法将120例患者随机分为试验组和对照组,每组60例,所有患者在积极治疗原发疾病同时,给予百普力采用持续输注的鼻饲方式进行肠内营养,试验组在肠内营养第1天开始给予神阙穴温和灸。结果在肠内营养治疗第7天时观察患者的腹泻发生率,发现试验组、对照组腹泻发生率分别为16.7%、36.7%,试验组明显低于对照组,经x2检验,P＜0.05,差异有统计学意义。结论神阙穴艾灸用于防治ICU危重患者肠内营养并发的腹泻疗效显著,且操作简便,成本低廉,值得推广应用。     

M-CSF、CA153、HPV联合检测在宫颈癌早期筛查中的临床意义

目的研究巨噬细胞集落刺激因子(M-CSF)、糖类抗原153(CA153)、人乳头瘤病毒(HPV)联合检测在宫颈癌早期筛查中的临床意义。方法以该院2016年3月至2018年10月收治的120例宫颈癌患者作为宫颈癌组,另选取同期于该院体检合格者120例作为健康组以及不典型增生患者120例作为CIN组。比较3组研究对象M-CSF、CA153、HPV水平差异,分析宫颈癌患者的M-CSF、CA153、HPV水平相关性以及单独检测和联合检测的效能差异。结果宫颈癌组患者的M-CSF、CA153、HPV水平均显著高于CIN组和健康组(P<0.05);宫颈癌组患者的M-CSF、CA153、HPV水平与宫颈癌呈正相关,均为宫颈癌的独立危险因素,通过ROC曲线分析,宫颈癌组患者的M-CSF、CA153、HPV联合检测的诊断灵敏度高于单独检测,宫颈癌患者的M-CSF、CA153的诊断界值分别为485.12pg/mL和317.62U/mL。结论M-CSF、CA153、HPV联合检测对宫颈癌早期筛查具有重要意义,在一定程度上可对患者的宫颈癌病变进行早期预测。