Synthesis of a novel series of L-isoserine derivatives as aminopeptidase N inhibitors

A series of novel L-isoserine derivatives were synthesised and evaluated for their ability to inhibit aminopeptidase N (APN)/CD13. In our preliminary biological results, some of these compounds possessed a potent inhibitory activity against the APN. Within this series, compound 14b not only showed similar enzyme inhibition (IC?? of 12.2 μM) compared with the positive control bestatin (half maximal inhibitory concentration (IC??) of 7.3 μM), but also had a potent antiproliferative activity against human cancer cell lines cells.     

基于MaxEnt模型预测中国兰属植物的分布格局及主导气候因子(附表)

兰属(Cymbidium)中，除了兔耳兰C.lancifolium以外的所有种均被列为国家重点保护野生植物。为探究其在未来气候条件下的潜在分布格局，该研究基于兰属植物已知的分布点和19个气候因子，利用最大熵(MaxEnt)模型和地理信息系统(ArcGIS)模拟兰属以及其中20种兰属植物在9种不同气候情景(当代以及未来2030s、2050s、2070s和2090s 4个时间段各两种温室气体排放情景)下的潜在分布格局。结果表明：(1)最干旱季降水量(Bio17)、年降水量(Bio12)和温度季节性变化(Bio4)是影响兰属植物地理分布格局的主导气候因子。(2)不同兰属植物在未来情景下的适生区表现出不同的变化趋势，并且影响其分布的主导气候因子也有所不同。其中，冬凤兰(C.dayanum)等8个物种的适生区面积整体呈扩张趋势，而西藏虎头兰(C.tracyanum)等12个物种的适生区面积整体则呈缩减趋势。该研究结果为兰属植物就地保护与迁地保护提供了重要参考，对兰属等濒危野生植物的保护具有积极意义。     

Formalin removal from archival tissue by critical point drying

The extraction of high-quality nucleic acid may be problematic in formalin-fixed tissues because of cross-linking between proteins and DNA. Old fixed tissue specimens do produce fragmented DNA (<1.2 kb), which is only used for PCR amplification. Here we show that high molecular weight DNA (>194 kb) can be successfully extracted from fixed tissue samples (16-70 years old) by gradual dehydration and critical point drying. The reliability of extracted DNA was measured by its ability to serve as a template for the amplification of mtDNA fragments (403 and 1198 bp) and an nDNA fragment (1844 bp). In addition, fingerprinting analysis was performed using DNA from fixed human tissue to ensure the ability of extracted DNA to hybridize with the DNA probe. DNA derived by this method can be subject to amplification, complete digestion by restriction endonuclease, and hybridization.     

Screening of high α-arbutin producing strains and production of α-arbutin by fermentation

A mutant Xanthomonas maltophilia BT-112 with high α-anomer-selective glycosylation activity was screened by a series of mutation methods including UV light, N-methyl-N-nitro-N-nitroso-guanidine treatment and quick neutron mutation. The α-arbutin titer increased 15-folds compared with the parent strain. The optimal conditions for culture medium and the operational conditions for lab-scale fermenter were investigated. Under optimized conditions, the maximal hydroquinone (HQ) tolerance of cells and yield of α-arbutin were 120 mM and 30.6 g/l, respectively. The molar conversion yield of α-arbutin based on the amount of HQ supplied reached 93.6 %. The product was identified as α-arbutin by ¹³C NMR and ¹H NMR analysis. In conclusion, the results in this work provide a one-step and cost-effective method for the large-scale production of α-arbutin.     

不同林龄油松(Pinus tabulaeformis)人工林植物、凋落物与土壤C、N、P化学计量特征

在陕西省北部延安市境内子午岭林区,采用时空互代的方法选取9、23、33、47年生油松(Pinus tabuliformis)人工林为研究对象,比较油松不同器官(叶、枝、干、根)、凋落物及土壤C、N、P含量及其比值的差异,探讨它们随林龄的变化及其相互间的关系,以期为油松人工林的生产、改善和林木生长环境的调节提供参考。结果表明:除根中C含量在林龄间差异不显著外,其它器官C、N、P含量及其比值在林龄间均差异显著且随林龄增加变化趋势不尽相同。9、23、33、47年生油松林C、N、P含量及N∶P比值均在叶中最高;C∶N比值均在干中最高,根中次之;C∶P比值均在干中最高,其它器官大小次序不一。除33年生油松林叶中N∶P比值大于14外,其它各器官各林龄N∶P比值均小于14,且N∶P比值随林龄先增加后减少,故可判断油松在该区域受N限制较为严重,且随林龄的增加受N限制的情况有所缓解。不同林龄土壤和凋落物C、N、P含量及其比值差异显著,且后者均大于前者。土壤与凋落物C、P含量及C∶N、C∶P、N∶P比值随林龄增加变化趋势完全一致,表明土壤与凋落物之间有着密切的关系。叶片与凋落物N、P含量及C∶N、C∶P、N∶P比值之间显著相关,表明凋落物的养分承自植物叶片,二者之间关系紧密;植物和土壤的C、N、P含量之间均不存在显著相关性,说明土壤C、N、P供应量对乔木叶片C、N、P含量影响不大。     

Mutations in the trfA replication gene of the broad-host-range plasmid RK2 result in elevated plasmid copy numbers.

Mutated forms of trfA, the replication protein gene of plasmid RK2, that support a minimal RK2 origin plasmid in Escherichia coli at copy numbers up to 23-fold higher than normal have been isolated. Six such high-copy-number (copy-up) mutations were mapped and sequenced. In each case, a single base transition led to an amino acid substitution in the TrfA protein primary sequence. The six mutations affected different residues of the protein and were located within a 69-base-pair region encoding 24 amino acids. Dominance tests showed that each of the mutants can be suppressed by wild-type trfA in trans, but suppression is highly dependent on the amount of wild-type protein produced. Excess mutant TrfA protein provided in trans significantly increased the copy number of RK2 and other self-replicating derivatives of RK2 that contain a wild-type trfA gene. These observations suggest that the mutations affect a regulatory activity of the TrfA replication protein that is a key factor in the control of initiation of RK2 replication.     

葡萄糖对ICR小鼠胚胎体外发育的影响

研究葡萄糖在小鼠早期胚胎体外发育中的作用。实验1将6—8周龄的ICR雌鼠超数排卵后与公鼠交配,收集1-细胞放入含0(对照组)、0.5、1、3、5、10mmol/L葡萄糖的CZB中培养;实验2将从超排的ICR雌鼠输卵管内收集的1-细胞放入无糖CZB中培养,分别于1细胞、2细胞、4细胞、桑椹胚阶段移入含3.0mmol/L葡萄糖(最适浓度)的CZB中,培养24h后又移回到无糖CZB中(桑椹胚阶段除外)继续培养以及整个胚胎培养过程均在含糖CZB中,对照组胚胎培养全程均在无糖CZB中。每组胚胎于37℃、5%CO2培养箱中培养120h,每24h在倒置显微镜下观察胚胎发育情况,分别计算2-细胞率、4-细胞率、桑椹胚率、囊胚率和孵化率,并进行囊胚细胞计数。结果显示,小鼠胚胎在含糖CZB中与在无糖CZB中4-细胞发育率无差异;含糖CZB中囊胚率显著高于对照组;3.0mmol/L浓度组囊胚细胞数显著高于其余组;2-细胞至4-细胞、4-细胞至桑椹胚前添加葡萄糖囊胚率显著高于对照组,1-细胞至2-细胞、桑椹胚及其以后阶段添加葡萄糖囊胚率与对照组无差异。实验证实,在ICR小鼠胚胎体外培养中加入葡萄糖不会导致2-细胞阻滞;葡萄糖浓度增至10mmol/L对ICR小鼠胚胎无毒性作用;ICR小鼠胚胎体外培养的最适葡萄糖浓度为3.0mmol/L;2-细胞至4-细胞、4-细胞至桑椹胚前添加葡萄糖是必要的。     

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.     

兔脑微栓塞模型脑血流动力学的CT灌注动态变化

目的探讨兔脑微栓塞模型CT灌注成像（CT perfusion imaging,CTPI）脑血流动力学的动态变化规律。方法 30只新西兰兔,随机分成两组,A组：假手术对照组5只,B组：微栓塞组25只。经颈外动脉向颈内动脉注入直径约0.5 mm的SiO2颗粒10枚,分别于栓塞后30 min、3 h、6 h、12 h及24 h行CTPI,24 h处死动物取脑组织行HE染色。根据HE染色结果将模型分为缺血组和梗死组,分别观察其脑血流量（cerebral blood flow,CBF）、脑血容积（cerebral blood volume,CBV）和平均通过时间（mean transit time,MTT）的动态变化规律。结果 A组CTPI及HE染色均未见明显异常。B组3只因实验意外死亡,1只因下肢静脉穿刺失败导致CTPI失败,21只行CTPI,其中18只灌注异常,3只未见明显异常。18只灌注异常的兔中,HE染色10只脑梗死,7只脑缺血,1只未见明显异常。30 min时7只缺血兔脑不同程度低灌注,表现为CBF降低,MTT延长,CBV无显著变化,3～6 h低灌注进一步加重,CBV值略降低,12 h低灌注不同程度恢复,24 h进一步恢复。30 min时10只梗死兔脑明显低灌注,表现为CBF及CBV显著降低,MTT显著延长,3只兔低灌注分别在3 h、6 h及12 h不同程度恢复,然后下一时间又迅速降低并随着时间延长进一步加剧,其余7只兔低灌注程度随时间延长逐渐加剧或在一定水平上波动。结论脑缺血3～6 h低灌注最明显,12～24 h低灌注不同程度恢复,而脑梗死随时间延长低灌注程度不断加重或一过性恢复后再次加重。脑缺血的特征是CBF和CBV的不匹配,缺血组织CBF显著降低,CBV无显著变化,而脑梗死则表现为这两个参数的一致性下降。