硫酸乙酰肝素协同细胞因子对脐血造血细胞体外扩增及细胞免疫表型的影响

目的:探讨硫酸Z酰肝素(HS)协同细胞因子对人脐血造血细胞体外生长的影响.方法:分成四组.A组用细胞因子(rIL3.rIL-lβ.rIL-6,和SCF等)、B组用骨髓基质细胞培养上清(SN) 细胞因子、C组用SN、D组用HS 细胞因子体外培养扩增脐血造血细胞.并应用间接免疫荧光法检测细胞表面标志.结果:①培养7 d.细胞已开始增殖,14 d达高峰,细胞总数A、B、C、D四组依次增加了134.5、171.3、81.5、167.2倍.2l d细胞数量开始下降,D组下降最明显.35 d时细胞总数依次仅为培养前的54.15、92.25、26.65和100.2倍.②CD34 细胞第7 d已升高,14 d达高峰,依次比培养前增加了68.43%、82.47%、69.77%和79.3%,21 d开始减少,但仍依次比培养前增加56.16%、71.72%、12.33%和73.7%.CD33 细胞7 d时均显著增加,14 d达高峰,依次比培养前增加了80.22%、68.64%、81.58%和70.30%,21 d时维持在14 d水平.CD38 细胞7 d时均有显著增加.比培养前增加了66.65%、73.8%、70.42%和71.9%,14～21 d均在该水平上维持.结论:①骨髓基质细胞培养上清与细胞因子联用对脐血造血细胞增殖的促进作用明显强于两者单用.②硫酸乙酰肝素可替代骨髓基质细胞培养上清与细胞因子联用.     

骨髓间充质干细胞与上清液治疗大鼠脑梗死的疗效

目的探讨骨髓间充质干细胞与上清液治疗大鼠脑梗死的疗效。方法将40只脑梗死大鼠模型用随机数字表法分为氯化钠注射液组(9 g.L-1氯化钠注射液2 mL)、骨髓间充质干细胞(BMSCs)移植组(2.0×109L-1BM-SCs悬液2 mL)、上清液移植组(BMSCs上清液2 mL)及混悬液移植组(上清液与BMSCs混悬液2 mL),每组10只。在模型再灌注24 h后,用10 g.L-15-溴脱氧尿核苷(BrdU)溶液标记,连续标记28 d,并于最后1次注射后2 h处死大鼠。观察各组病灶增殖期神经前体细胞的数目变化及神经损伤严重程度(NSS)评分。结果治疗后28 d BrdU、Br-dU+抗人神经元特异性烯醇化酶、BrdU+胶质纤维酸性蛋白标记的阳性细胞数BMSCs移植组、上清液移植组和混悬液移植组较氯化钠注射液组明显增多,差异有统计学意义(P<0.05);上清液移植组和混悬液移植组较BMSCs移植组增多,差异有统计学意义(P<0.05);混悬液移植组较上清液移植组增多,差异有统计学意义(P<0.05)。与移植前比较,各组大鼠动物模型移植后第7、14、28天的神经功能情况均有不同程度的恢复,NSS评分逐渐降低,差异均有统计学意义(P<0.05或P<0.01);从第7天起,与氯化钠注射液组比较,BMSCs移植组、上清液移植组和混悬液移植组NSS评分均明显降低,差异有统计学意义(P<0.05或P<0.01);混悬液移植组NSS评分明显低于BMSCs移植组和上清液移植组,差异有统计学意义(P<0.01)。结论上清液与BMSCs混悬液能够诱导病灶区内源性神经前体细胞的增殖、分化,这可能与上清液内的神经营养因子及细胞分化诱导因子诱导BMSCs分化为神经细胞有关。     

Biologic characteristics of an immunosuppressive factor derived from a human lung cancer cell line

A 549, a human lung cancer cell line, spontaneously produces a tumor-derived immunosuppressive factor (TDSF) which inhibited PHA-stimulated T lymphocyte proliferation via a noncytotoxic mechanism. The inhibition increased in a dose-dependent pattern. The factor also markedly suppressed production of interleukin (IL-2) by PHA-stimulated lymphocytes and IL 2-dependent proliferation of activated lymphocytes. The fact that TDSF possessed very potent inhibitive action on IL-2 is especially noteworthy if we consider the use of IL-2 as immunotherapeutic agent. The synthesis of the factor was inhibited by mitomycin C, actinomycin D and cycloheximide, indicating that the factor is a genic product of A 549 cells. The factor is chemically a protein with a molecular weight greater than 150 KD and sensitve to extremes of pH, heating to 60 °C and trypsin treatment.     

骨髓间充质干细胞上清液对大鼠佐剂性关节炎防治作用的实验研究

目的 探讨骨髓间充质于细胞(mesenchymal stem cells,MSCs)上清液对大鼠佐剂性关节炎(adjuvant arthritis,AA)的治疗、干预作用.方法 收集培养48 h的SD大鼠骨髓MSCs上清液,健康雄性SD大鼠右后足跖皮内一次性注射弗氏完全佐剂0.1 mL/只造模,造模成功后行MSCs上清液治疗及干预.造模后第1、7、14、21和28天测量大鼠体质量、踝关节周长和后足爪体积,同时观察大鼠生存状况.治疗后第21天,行四肢关节x线检查及病理检查.结果 模型大鼠经MSCs上清液治疗后关节肿胀明显减轻,X线示骨质破坏明显修复.病理HE染色见关节滑膜病变和炎症细胞浸润明显减少,组织水肿减轻.干预组经MSCs上清液治疗,原发性炎症和继发病变均不明显,关节X线和病理检查均未见明显异常,与模型组大鼠相比,体质量、足爪体积以及踝关节周长差异有统计学意义(P＜0.05).结论 MSCs上清液治疗可有效减轻并逆转弗氏完全佐剂诱导的大鼠AA的进展.其作用机制可能为MSCs分泌了可溶性细胞因子到达患部修复受损组织,具体成分还有待进一步研究.     

β-榄香烯对肿瘤细胞上清液诱导的大鼠骨髓来源内皮祖细胞增殖与凋亡的影响

目的观察β-榄香烯对肿瘤细胞上清液诱导的大鼠骨髓来源内皮祖细胞(EPCs)增殖与凋亡的影响,探讨β-榄香烯对EPCs参与肿瘤血管形成过程的作用。方法原代培养大鼠骨髓来源EPCs经胃腺癌细胞株SGC-7901培养上清液诱导EPCs 12h后,分为浓度效应组(β-榄香烯0、5、10、20mg.L-1干预48h)和时间效应组(20mg.L-1β-榄香烯干预0、12、24、48h)。检测各组EPCs的增殖和凋亡率。结果随着β-榄香烯剂量的增加和作用时间的延长,EPCs的增殖显著下降(P<0.05),当β-榄香烯浓度为5mg.L-1及作用时间为12h时,EPCs凋亡率变化不明显,当β-榄香烯浓度增加至10mg.L-1及作用时间延长为24h时,EPCs凋亡率显著上升(P<0.05)。结论β-榄香烯通过诱导EPCs的凋亡抑制其增殖,并可削弱EPCs在肿瘤血管发生中的作用。     

Characterization of synthetic medium antigens of Micropolyspora faeni and Thermoactinomyces candidus

A synthetic (Syn) medium was developed for growth of Micropolyspora faeni and Thermoactinomyces candidus, and optimum conditions for culture filtrate antigen production were found to be 3 days for T. candidus and 9 to 12 days for M. faeni. Appearance of proteolytic activity in the culture fluid supernatant coincided with a decrease in precipitating antigen content, protein content, and number of proteins on polyacrylamide gel electrophoresis (PAGE), probably as a result of proteolysis. The results suggest that protein content, number of protein bands on PAGE, and proteolytic activity are predictive of precipitating antigen content. Antigens prepared in Syn medium were compared to those produced by the double-dialysis procedure and found to contain adequate amounts of antigenic material of sufficient quality to warrant clinical studies of their usefulness in the diagnosis of hypersensitivity pneumonitis.     

人胎垂体体外刺激胎脾淋巴细胞增殖的研究

采用ＭＴＴ法测定人胎儿垂体培养上清刺激胎脾淋巴细胞增殖反应。实验中观察到：（１）ＦＰＳ能刺激胎脾淋巴细胞增殖，其刺激能力随胎龄增大而增强；（２）胎儿垂体体能 在体外培养１ｍｏ内持续分泌对淋巴细胞增殖的有效刺激成分；（３）ＦＰＳ能延长胎脾淋巴细胞外培养存活时间；（４）ＰＨＡ对ＦＰＳ刺激胎脾淋巴细胞增殖无影响。实验结果显示ＦＰＳ对人胎脾淋巴细胞在体有刺激增殖作用。     

成纤维细胞上清提取物对成纤维细胞增殖迁移的影响

目的：在人皮肤成纤维细胞培养上清液中寻求出有效的生长因子样活性组分。方法：收集超滤成纤维细胞的培养上清液，采用MTT法、划痕试验等检测其对人皮肤成纤维细胞增殖和诱导迁移的影响。结果：上清液组中分子量10～30kD的试验组对成纤维细胞有明显促增殖趋势，而大于30kD组有较强的促进单层细胞伤口愈合的能力，二者与对照组比较均差异显著（P〈0．01）。结论：人成纤维细胞培养上清液的提取物中含有促进成纤维细胞增殖和迁移能力的活性物质。     

Current and future trends in non–small cell lung cancer biomarker testing: The American experience

Biomarker testing in patients with advanced stage non–small cell lung cancer provides essential information that can be used to select the most appropriate therapy. The regular updates of guideline recommendations reflect the growing number of biomarkers that must be assessed, and as such signal the shift from single-gene assays to more comprehensive genomic profiling using next-generation sequencing modalities. Cytology and small biopsy specimens have proven to be more than adequate substrates for these types of ancillary molecular testing; however, other alternative testing substrates are beginning to emerge. These include so-called liquid biopsies as well the supernatant fluid from cytology specimens, both of which have demonstrated promise for use in the clinical realm. This review will briefly cover the current state of non–small cell lung cancer biomarker testing in the United States, with a focus on these novel nonconventional substrates that are increasingly being incorporated into testing paradigms.