柱前衍生-RP-HPLC法测定青蒿中青蒿素的含量

采用柱前衍生-RP-HPLC法测定10个不同产地的青蒿药材中青蒿素的含量.采用Lichrospher 100 RP-18e(250 mm×4.6 mm,5μm,Merck KgaA,Germany)色谱柱,甲醇-0.01 mol/L醋酸钠-醋酸缓冲液(pH 5.8)(体积比62:38)为流动相;检测波长:260 nm;流速:0.5 mL/min;柱温:25℃.结果表明该法准确重现性好,可以为青蒿质量标准的制订提供科学依据.     

栓皮栎林的生物量

本文对北京西山海拔300米的26年生人工栓皮栎林的生物量作了测定。粮据十株伐倒的标准木分别取得乔木各部分的干重。采用W=a(D2H)b的幂函数方程,获得每公顷土地上的树干重32.16吨,枝条重9.85吨、叶重1.68吨、根重9.95吨。对灌木和草本植物采用收割法作测定：灌木的地上部分每公顷为1.33吨,地下部分为2.3吨;草本植物的地上部分每公顷为0.15吨,地下部分为0.8吨。所以栓皮栎的地上部分总生物量每公顷为45.17吨,地下部分13.05吨,总计58.22吨。     

九种蹄盖蕨科植物配子体的酯酶同工酶分析

本文培养了蹄盖蕨科９个种即中华蹄盖蕨（Ａｔｈｙｒｉｕｍｓｉｎｅｎｓｅ）带岭蹄盖蕨（Ａ．ｄａｌｉｎｇｅｎｓｅ）、多齿蹄盖蕨（Ａ．ｍｕｌｔｉｄｅｎｔａｔｕｍ）（包括青柄和紫柄２种带岭蹄蕨类型）、东北蛾眉蕨（Ｌｕｎａｔｈｙｒｉｕｍｐｙｃｎｏｓｏｒｕｍ）、朝鲜介蕨（Ｄｒｙｏａｔｈｙｒｉｕｍｃｏｒｅａｎｕｍ）、山冷蕨（Ｃｙｓｔｏｐｔｅｒｉｓｓｕｄｅｔｉｃａ）、假冷蕨（Ｐｓｅｕｄｏｃｙｓｔｏｐｔｅｒｉｓｓｐｉｎｕｌｏｓａ）、欧洲羽节蕨（Ｇｙｍｎｏｃａｒｐｉｕｍｄｒｙｏｐｔｅｒｉｓ）和黑鳞短肠蕨（Ａｌａｎｔｏｄｉａｃｒｅｎａｔａ）的配子体为材料,进行酯酶同工酶的分析,以表明它们的种间差异,其中多齿蹄盖蕨的青柄和紫柄两种类型的酶谱存在明显差异,应考虑紫柄为多齿蹄盖蕨的变型。     

Engineering imaging probes and molecular machines for nanomedicine

Nanomedicine is an emerging field that integrates nanotechnology, biomolecular engineering, life sciences and medicine; it is expected to produce major breakthroughs in medical diagnostics and therapeutics. Due to the size-compatibility of nano-scale structures and devices with proteins and nucleic acids, the design, synthesis and application of nanoprobes, nanocarriers and nanomachines provide unprecedented opportunities for achieving a better control of biological processes, and drastic improvements in disease detection, therapy, and prevention. Recent advances in nanomedicine include the development of functional nanoparticle based molecular imaging probes, nano-structured materials as drug/gene carriers for in vivo delivery, and engineered molecular machines for treating single-gene disorders. This review focuses on the development of molecular imaging probes and engineered nucleases for nanomedicine, including quantum dot bioconjugates, quantum dot-fluorescent protein FRET probes, molecular beacons, magnetic and gold nanoparticle based imaging contrast agents, and the design and validation of zinc finger nucleases (ZFNs) and TAL effector nucleases (TALENs) for gene targeting. The challenges in translating nanomedicine approaches to clinical applications are discussed.     

Inhibition of mitochondrial complex I improves glucose metabolism independently of AMPK activation

Accumulating evidences showed metformin and berberine, well‐known glucose‐lowering agents, were able to inhibit mitochondrial electron transport chain at complex I. In this study, we aimed to explore the antihyperglycaemic effect of complex I inhibition. Rotenone, amobarbital and gene silence of NDUFA13 were used to inhibit complex I. Intraperitoneal glucose tolerance test and insulin tolerance test were performed in db/db mice. Lactate release and glucose consumption were measured to investigate glucose metabolism in HepG2 hepatocytes and C2C12 myotubes. Glucose output was measured in primary hepatocytes. Compound C and adenoviruses expressing dominant negative AMP‐activated protein kinase (AMPK) α1/2 were exploited to inactivate AMPK pathway. Cellular NAD⁺/NADH ratio was assayed to evaluate energy transforming and redox state. Rotenone ameliorated hyperglycaemia and insulin resistance in db/db mice. It induced glucose consumption and glycolysis and reduced hepatic glucose output. Rotenone also activated AMPK. Furthermore, it remained effective with AMPK inactivation. The enhanced glycolysis and repressed gluconeogenesis correlated with a reduction in cellular NAD⁺/NADH ratio, which resulted from complex I suppression. Amobarbital, another representative complex I inhibitor, stimulated glucose consumption and decreased hepatic glucose output in vitro, too. Similar changes were observed while expression of NDUFA13, a subunit of complex I, was knocked down with gene silencing. These findings reveal mitochondrial complex I emerges as a key drug target for diabetes treatment. Inhibition of complex I improves glucose homoeostasis via non‐AMPK pathway, which may relate to the suppression of the cellular NAD⁺/NADH ratio.     

Expression of a cytotoxic cationic antibacterial peptide in Escherichia coli using two fusion partners

It has been reported that it is difficult to express cationic antibacterial peptides in engineered bacteria because such peptides are highly toxic to the host bacteria cells and sensitive to intracellular proteases. Antibacterial peptide CM4 (ABP-CM4) is a small cationic peptide with broad-spectrum activities against bacteria, fungi and tumor cells, which may possibly be used as an antimicrobial agent. Here we tried to express ABP-CM4 in Escherichia coli cells using either the GST fusion system or the intein-mediated fusion expression system. In order to investigate the possible use of these two fusion partners in cationic small peptide expression and purification, a mutant ABP-CMt, which is a highly positively charged peptide with +9 charges at neutral pH, was designed. In the present study, we have shown that both ABP-CM4 and ABP-CMt peptides can be expressed and purified by the intein-mediated expression system but not by the GST fusion expression system. Thus the intein-mediated peptide expression and purification system potentially could be employed for the production of recombinant protease-sensitive and cytotoxic peptides.     

博尔纳病病毒抗原免疫的杂交瘤细胞系的评价

目的评价博尔纳病病毒(BDV)抗原免疫的杂交瘤细胞系的产生抗体能力与抗体特异性。方法随机抽取2株由BDV抗原免疫的杂交瘤细胞系H1和H2,通过体内和体外方法制备单克隆抗体,通过间接免疫荧光、免疫印迹和间接酶联免疫吸附3种试验方法对制备的抗体进行特异性鉴定及效价测定。结果杂交瘤细胞系H1和H2可以产生一定效价的单克隆抗体,并对BDV的P24和P40抗原具有特异性。结论由杂交瘤细胞系H1和H2产生的单克隆抗体可以用于检测BDV特异性抗原。     

濒危植物明党参与非濒危种峨参种子休眠和萌发比较

研究了濒危植物明党参（Changium smyrnioides)与非濒危种峨参（Anthriscus sylvestris)种子贮存,打破休眠和萌发对水分和温度条件的要求。结果表明,在自然条件下,明党参种子有5个月的休眠期,人工低温（10℃左右）处理40天即可打破休眠;两种植物种子自然温度干燥处理不能打破休眠;两种植物的种子在自然温度变湿层积处理后萌发率最高,萌发持续时间也最长,其中明党参的萌发率地峨参,持续时间短于峨参;自然温度淹水处理大大降低了两种植物种子的萌发率,但明党参仍有7％的萌发率。明党参种子质量和发芽率不应是明党参濒危的直接原因,但因其具有种子产量低,幼苗数量少,存活率高的K－对策,当受到强烈干扰时,种群难以在短期内恢复,容易濒危。     

大渡河上游不同林龄云杉人工林与原始林下地表苔藓层片结构与物种组成

为评估在人工林早期发育过程中地表苔藓层片物种组成和结构的演变趋势, 选择四川省金川县507林场4–30年林龄的5块云杉(Picea asperata)人工林和1块300年岷江冷杉(Abies faxoniana)原始林, 开展了地表苔藓植物调查。采用方差分析法(ANOVA)对苔藓植物特征参数进行差异性检验, 采用Sørensen群落相似性系数比较了苔藓群落β多样性差异。结果表明: (1)云杉人工林较原始林地表苔藓物种丰富度高。年轻的人工林(<16年)较中龄林(21–30年)地表有更多的苔藓种类; (2)原始林较人工林有更高的地表苔藓植物盖度、密度、平均高度和厚度, 而不同林龄的人工林之间在苔藓植物层片盖度和密度上并没有表现出统计学意义上的差异(P>0.05), 但地表苔藓优势种及次优势种组成具有较明显的差异; (3)人工林与原始林地表苔藓共有种具有明显的喜光耐旱特性, 4年生未成林地段与原始林共有种数最多, 为19种; (4)原始林下地表24种苔藓植物中, 除Rhytidium rugosum外, 23种在云杉人工林早期发育过程中存在。在人工林发育过程中地表苔藓物种替代明显, 替代率随林龄呈增加趋势(0.24–0.60), 有明显的物种替代现象发生。综合分析表明: (1) 本文所研究的4–30年的云杉人工林发育过程中地表苔藓结构与多样性并没有表现出预期的恢复趋势, 大多数土著苔藓种群还未能有效恢复; (2)要恢复重建林地后演替阶段的苔藓种群结构和多样性, 不仅需要在采伐和造林过程中减少对土壤的扰动以保护地表微生境, 还应在云杉人工林处于16–20年林龄, 地表苔藓植物丰富度发生显著衰退时进行合理疏伐。