凉水国家自然保护区天然红松种群遗传多样性在时间尺度上变化的cpSSR分析

应用叶绿体微卫星（cpSSR）技术对凉水国家自然保护区天然红松种群的遗传多样性在时间尺度上的变化进行了遗传分析。共选择121个树龄在1 581~1 900年间的红松个体,连续分为6个龄级。从6对叶绿体SSR引物筛选出两对引物,共检测到7个位点,其中多态位点5个,多态位点比率为71.43%。利用POPGENE分析软件计算了红松各龄级内的遗传多样性指数,分析了遗传多样性在红松各龄级间的变化。研究结果表明,凉水保护区内的红松遗传多样性在这320年间没有发生太大的波动,龄级间遗传多样性分化不明显,红松遗传多样性主要存在于龄级内。     

Role of calcium mobilization in the regulation of spontaneous transient outward currents in porcine coronary artery myocytes

The purpose of the present study was to further study the characteristics and regulation of spontaneous transient outward currents (STOCs) in freshly isolated porcine coronary artery smooth muscle cells (ASMCs). STOCs were recorded using the perforated whole-cell patch-clamp configuration. STOCs were voltage-dependent and superimposed stochastically onto whole-cell Ca2 -activated-K (BKCa) currents. Charybdotoxin (ChTX, 200 nmol/L), a selective blocker of BKCa channels, completely inhibited STOCs within 10 min. STOCs activity was greatly suppressed when extracellular Ca2 concentration decreased from 1.8 mmol/L to 200 nmol/L, further removal of Ca2 abolished STOCs activity. Ca2 ionophore A23187 (10 μmol/L) increased STOCs activity significantly. Verapamil (20 μmol/L) and CdCl2 (200 μmol/L), two kinds of organic L-type voltage-dependent Ca2 channels (L-VDCCs) antagonists, had little effect on STOCs. In addition, the ryanodine receptors (RyRs) agonist caffeine (5 mmol/L) significantly activated STOCs. Application of ryanodine (50 μmol/L) to block RyRs abolished STOCs, subsequent washout of ryanodine or application of caffeine failed to reproduce STOCs activity. Inhibition of inositol 1,4,5-trisphosphate receptors (IP3Rs) by 2APB (40 μmol/L) greatly suppressed the activity of STOCs, application of caffeine (5 mmol/L) in the presence of 2APB caused a burst of outward currents followed by inhibition of STOCs. These results suggest that STOCs in porcine coronary ASMCs are mediated by BKCa channels. Extracellular Ca2 is essential for STOCs activity, while Ca2 entry through L-VDCCs has little effect on STOCs. Intracellular Ca2 release induced by RyRs is responsible for the regulation of STOCs, whereas IP3Rs might also be involved.     

Full-thickness tissue engineered skin constructed with autogenic bone marrow mesenchymal stem cells

To explore the feasibility of repairing clinical cutaneous deficiency, autogenic bone marrow mesenchymal stem cells (BMSCs) were isolated and differentiated into epidermal cells and fibroblasts in vitro supplemented with different inducing factors and biomaterials to construct functional tissueengineered skin. The results showed that after 72 h induction, BMSCs displayed morphologic changes such as typical epidermal cell arrangement, from spindle shape to round or oval; tonofibrils, melanosomes and keratohyaline granules were observed under a transmission electronic microscope. The differentiated cells expressed epidermal stem cell surface marker CK19 (59.66% ± 4.2%) and epidermal cells differentiation marker CK10. In addition, the induced epidermal cells acquired the anti-radiation capacity featured by lowered apoptosis following exposure to UVB. On the other hand, the collagen microfibrils deposition was noticed under a transmission electronic microscope after differentiating into dermis fibroblasts; RT-PCR identified collagen type I mRNA expression in differentiated cells; radioimmunoassay detected the secretion of interleukin-6 (IL-6) and interleukin-8 (IL-8) (up to 115.06 pg/mL and 0.84 ng/mL, respectively). Further in vivo implanting BMSCs with scaffold material shortened skin wound repair significantly. In one word, autogenic BMSCs have the potential to differentiate into epidermal cells and fibroblasts in vitro, and show clinical feasibility acting as epidermis-like and dermis-like seed cells in skin engineering. Supported by the Major Technology Program of Beijing Municipal Science & Technology Commission (Grant No. H060920050130) and the Major State Basic Research Development Program of China (Grant No. 2005CB522702)     

小RNA深度测序技术分析西瓜花叶病毒蜀葵分离物

蜀葵病毒病害的发生对其生长造成严重影响,明确蜀葵病毒病害的种类及变异进化对蜀葵病毒病害的防治具有重要意义。利用小RNA深度测序技术对具有明显脉明、花叶症状的蜀葵叶片进行鉴定。结果发现,感病蜀葵被西瓜花叶病毒(Watermelon mosaic virus, WMV)、锦葵脉明病毒(Mala vein cleaning virus, MVCV)和一种新的RNA病毒[暂命名为蜀葵病毒1号(Althaea rosea virus1, ArV1)]所侵染。为进一步明确WMV蜀葵分离物(WMV-Tg)的进化关系,对病毒WMV-Tg全基因组进行扩增,获得全长为10 046个核苷酸序列(nt)。序列分析结果显示,WMV-Tg与已报道的WMV分离物基因组核苷酸序列的同源性为83.3%～90.2%。系统进化关系表明,WMV-Tg与WMV-Pg聚为一簇,亲缘关系最近。对蜀葵WMV-Tg来源的小RNA(WMV-derived small interfering RNAs, WMV-vsiRNAs)的长度分布、5′碱基偏好性、极性分布以及热点区分布的分析,有助于加深对WMV-vsiRNAs的了解,并为进一步研究病毒来源的小RNA(virus-derived small interfering RNAs, vsiRNAs)在抗病毒防御中的功能,以及为蜀葵病毒病的防治奠定理论基础。     

基于核酸适配体的肿瘤免疫治疗进展

肿瘤免疫治疗是通过调节机体的免疫功能来控制和杀伤肿瘤的一种治疗手段。针对免疫检查点的治疗等一系列临床突破使得肿瘤的免疫治疗受到了广泛重视。目前,抗体治疗和过继性细胞治疗是肿瘤免疫治疗的主要方式,但是这些方法仍具有副作用较强,实体瘤治疗难以实现,治疗费用高昂等缺点。因此改进和发展更加高效、安全、低成本的新技术仍十分必要。适配体是利用指数富集的配体系统进化技术筛选得到的单链寡核苷酸,有核酸"抗体"之称。适配体具有低免疫原性、组织穿透力强、易于化学合成与修饰等优势,且与其靶标的结合具有较好亲和力和特异性,可像抗体一样实现肿瘤的免疫治疗。对适配体在肿瘤免疫治疗相关技术中的新应用作一综述,主要包括基于免疫检查点的抗肿瘤作用、双特异性适配体的肿瘤免疫治疗、适配体靶向递送siRNA的肿瘤免疫治疗和适配体联合抗体的肿瘤免疫治疗等方面。     

锁阳中氨基酸类化合物研究进展

锁阳（Cynomorium songaricum）是寄生于白刺属（Nitraria L.）植物根部的肉质草本植物,含有多种生物活性成分,如黄酮类、萜类、氨基酸等。氨基酸是锁阳中的一类重要化合物,与锁阳的品质密切相关,具有较大的研究价值。本文通过查阅近几年的相关文献,从锁阳中含有的氨基酸种类、含量、生物活性、检测方法及人工培育前景等方面对锁阳中氨基酸类成分的研究现状进行整理和归纳,旨在为锁阳中氨基酸类成分的进一步研究、开发和利用提供一定的支持与帮助。     

危重病人预警蛋白质组指纹与肿瘤患者生存关系的报告

目的:探讨危重病人预警蛋白质(lost goodwill target,LGT)多肽谱指纹的峰型和丰度对判断肿瘤患者病情发展及预后的意义.方法:对接受SELDI(Surface enhanced laser desorption/ionization-time of flight-mass spectrometer)技术检测的178名患者生存期与死亡时间进行调查,并根据SELDI检测所获得的指纹图中,以LGT指纹存在与否,分成无峰组、单峰组、双峰组、三峰和多峰组.采用多因素方差分析和COX风险回归模型分析的方法,比较四组中哪一种状态下死亡风险最大.结果:恶性肿瘤患者经SELDI技术检测所获取的指纹图中,如果LGT呈阴性表达,死亡风险低于单、双峰组,远低于三峰和多峰组;LGT呈三峰和多峰表达,其死亡风险远高于单、双峰组,差异有统计学意义(P<0.05),死亡率极高.结论:采用SELDI技术检测肿瘤患者的血清,LGT出现三峰和多峰表达,提示肿瘤患者病情即将恶化,该指纹标识为LGT阳性指标;单、双峰者为疑似指标;LGT指纹阴性表达,肿瘤患者近期预后良好,死亡风险极小.     

环境酸化引起钙失衡的生理生态学效应

环境酸化是全球气候变化引起的主要问题之一,陆地和湖泊等生态系统受酸雨的长期影响,呈现出pH下降的趋势,而海洋生态系统也由于大气CO₂浓度的急剧上升而出现酸化。环境的酸化对生态系统中Ca的平衡造成巨大影响。在海洋生态系统中表现为,随着pH下降海水中CO₃^2-浓度降低,富Ca海洋生物无法维持其体内CaCO₃形成的硬质结构,造成珊瑚钙化生长变缓、翼足目软体动物外壳腐蚀穿孔等,影响其种群繁衍。另外,在陆地生态系统中,由于酸雨的长期淋溶,大量的生物可利用Ca从生态系统中流失,动植物无法获得充足的Ca来满足其生理需要。蜗牛、水虱等土壤及湖泊中高Ca动物的生长、繁殖受抑制,鸟类也由于缺Ca造成种群数量下降。陆地植物体内依赖于Ca的信号转导系统也遭受严重干扰,进而影响光合、抗逆和繁殖等诸多生理过程,表现出不同程度的生产力、物种多样性下降和森林的衰退。环境酸化导致不同生态系统中Ca循环失衡,大多表现出不利于富Ca生物体的存活,这可能是一个较为普遍的生态学模式。     

体外心肌细胞缺血再灌注损伤模型的建立

目的建立体外心肌细胞缺血再灌注(I/R)模型,为研究各种心血管疾病及药物治疗提供基础.方法体外培养新生SD乳鼠心肌细胞,实验分正常组和缺血再灌注组.倒置显微镜及透射电子显微镜下观察心肌细胞缺血再灌注前后的形态变化,MTT、流式细胞术检测细胞凋亡情况,免疫组化和Western Blot检测Bcl-2/Bax基因表达.结果 I/R前后心肌细胞形态发生明显变化.与正常组相比,I/R组心肌细胞凋亡率明显增加,细胞活性降低.免疫组化和Western Blot检测结果均出现I/R后Bcl-2蛋白表达下调,Bax蛋白表达上调,Bcl-2/Bax显著减小的现象.结论体外培养的心肌细胞I/R损伤各项指标检测与动物体内实验结果相吻合,体外心肌细胞缺血再灌注模型建立成功.