多基因模型在肝细胞癌预后中的应用

利用较少分子信息预测肝细胞癌类型对患者的个性化治疗十分关键。探索已知的与肝细胞癌预后相关的信号通路,共发现41个关键基因。随后,运用机器学习的方法对其构建风险预测模型,并在4个肝细胞癌数据集上进行验证。结果显示,该模型能将肝细胞癌患者分成两个预后差异显著的类型:癌症基因图谱(The cancer genome atlas,TCGA)数据集交叉验证的平均log rank P值为0.03;其他测试数据集的log rank P 值分别为0.000 38、0.002 1和0.01。生物信息学分析显示肝细胞癌的预后与细胞周期等信号通路显著相关,并筛选出12个潜在的肝细胞癌分子标志物。研究结果表明,基于41个基因构建的肝细胞癌预后模型具有较好的稳健性和准确的风险预测能力。     

条件启动子pCTR4在隐球菌基因表达调控中的应用

目的通过条件启动子pCTR4的质粒构建以及其在新生隐球菌中的同源置换,研究其在隐球菌基因表达调控中的应用。方法应用套叠PCR,构建含报告基因NEO的铜离子抑制性启动子质粒pNEO/CTR4和启动子同源重组框,并利用基因枪将其转化入新生隐球菌感受态细胞,常规及实时定量PCR检测条件启动子对目的基因的转录调控效应。结果成功构建了质粒pNEO/CTR和隐球菌条件启动子重建菌株,条件启动子pCTR4对目的基因具有预期的转录诱导和抑制效果。结论新建铜离子抑制性启动子质粒pNEO/CTR4可以应用于对隐球菌目的基因表达水平的调控;隐球菌泛素编码基因UBI 1并非致死性关键基因。我们的研究为今后新生隐球菌泛素系统的分子致病机制研究奠定了基础。     

Somatic embryogenesis and organogenesis from mature caryopses of North African barley accession “Kerkena” (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.)

The regeneration ability of a Tunisian barley accession originated from Kerkena islands was monitored through somatic embryogenesis and organogenesis. To prevent or to reduce normal germination, longitudinally bisected as well as base-wounded mature caryopses were cultured on a modified Chée and Pool-based medium (CP) enriched with different phytohormonal combinations. The greatest embryogenesis response was obtained when base-wounded caryopses were cultured on CP enriched with 2 mg/l chlorophenoxyacetic acid + 2.5 mg/l kinetin (76.85%). The same combination coupled to longitudinally bisected caryopses led to the embryogenic induction at the hypocotyl base of the germinated caryopses (61.9%). Embryogenic calluses differentiated into globular, heart-shaped, torpedo, and fully differentiated stages of somatic embryos on hormone-free Murashige and Skoog-based medium. Rooted plantlets were successfully transferred to soil and grown to maturity in the greenhouse and produced fertile seeds within 3 mo. On the other hand, organogenesis was achieved on CP enriched with 2 mg/l 2,4-dichlorophenoxyacetic acid + 2.5 mg/l kinetin. Histological aspects and scanning electron microscopy of both regeneration methods confirmed further the embryogenic and organogenic nature of the established processes. This efficient plant regeneration system provides a foundation for generating transgenic plants and germplasm preservation of “Kerkena” barley accession.     

Quaesitol,a phenol from Garcinia quaesita

The petrol extractives of the bark of Garcinia quaesita gave hermonionic acid, its decarboxylated product and a new phenol, quaesitol.     

Micromorphology,structural and ultrastructural changes during somatic embryogenesis of a Tunisian oat variety (<Emphasis Type="Italic">Avena sativa</Emphasis> L. var ‘Meliane’)

To better understand micromorphological and structural changes, histological sections provide additional insight into cellular process and developmental pathways occurring in oat somatic embryogenesis. Environmental scanning electron microscopy (ESEM) and transmission electron microscopy (TEM) were also used to follow the ultrastructural modifications during this system. Histological observations allowed following the events leading to the development of mature somatic embryos. The scheme includes the following steps: cell reactivation, the first organized cell division in diads, triads, tetrads as well as octant stages, the observation of an extracellular matrix (ECM) as a fibrillar material that bounded the surface of individualized proembryos. The transition from proembryo stage to an early globular somatic embryo was noted, where the embryogenic cortex is surrounded by the protoderm. The late globular stage was marked by bipolarity. The early and late transitional stages, the coleoptilar, mature and germinated stages were also described. The ESEM allowed us to follow some rearrangements, related to the morphology and surfaces involved in somatic embryos development. These events are proembryo formation, transition from proembryo to globular stage, marked by protoderm formation, scutellum and coleoptile development and finally somatic embryos germination. The TEM showed that embryogenic cells were very rich in organelles; mitochondria, rough endoplasmic reticulum, Golgi apparatus and ribosomes. Cells of proembryos, globular and late somatic embryos showed more vacuoles and differentiated organelles. The ECM was also detected by TEM as fibrillar material coating the cell walls. These results on structural and ultrastructural changes provided new insights and findings on oat somatic embryogenesis.     

The origin of interspersed repeats in the human genome

Over a third of the human genome consists of interspersed repetitive sequences which are primarily degenerate copies of transposable elements. In the past year, the identities of many of these transposable elements were revealed. The emerging concept is that only three mechanisms of amplification are responsible for the vast majority of interspersed repeats and that with each autonomous element a number of dependent non-autonomous sequences have co-amplified.     

Anomalous Cases of Conjugation and Spontaneous Autogamy in Stylonychia mytilus: A Note on the Asexuality of Early Conjugants

Four types of anomalous conjugation were documented in Stylonychia mytilus. Type I pairs were formed between mates of different sizes. These pairs exhibited an abnormal site of fusion in at least one of the mates, and the mates might face each other ventrally throughout conjugation instead of the normal side-by-side position. Type I pairs underwent sexual nuclear development and proceeded with the first cortical reorganization as in normal conjugants. Type II involved pairing at the anterior ends of mates with ventral surfaces facing the same direction. These pairs also underwent sexual nuclear development. Hence, aberrant orientation of the mates, and also ectopic sites of cytoplasmic fusion, if extensive, would permit sexual development. Type III pairs were united ventral-to-ventral with their anterior-left sides at the adoral zone of membranelles, and remained as such throughout conjugation. In these pairs, nuclear and cortical events were typical of the asexual development of physiological reorganization. In Type IV pairs, one mate of the pair possessed a fission furrow and developed two sets of ciliature typical of binary fission, while the other mate might undergo physiological reorganization or binary fission. Type III and Type IV pairs thus reveal the asexual state of early conjugants, which can pursue either one of the two modes of asexual cortical reorganization; these cases reinforce the notion of overlap of asexual and sexual cycles during conjugation of hypotrichs. Spontaneous autogamy was documented for the first time for this genus. The autogamonts proceeded with nuclear development and with the first cortical reorganization. Some probably underwent second and third reorganizations, as in conjugants, but accompanied by abnormalities, particularly in the stages beyond fertilization. Post-autogamous clones were nonviable except for one dubious case.     

Multicellular compartmentation of catharanthus roseus alkaloid biosynthesis predicts intercellular translocation of a pathway intermediate

In situ RNA hybridization and immunocytochemistry were used to establish the cellular distribution of monoterpenoid indole alkaloid biosynthesis in Madagascar periwinkle (Catharanthus roseus). Tryptophan decarboxylase (TDC) and strictosidine synthase (STR1), which are involved in the biosynthesis of the central intermediate strictosidine, and desacetoxyvindoline 4-hydroxylase (D4H) and deacetylvindoline 4-O-acetyltransferase (DAT), which are involved in the terminal steps of vindoline biosynthesis, were localized. tdc and str1 mRNAs were present in the epidermis of stems, leaves, and flower buds, whereas they appeared in most protoderm and cortical cells around the apical meristem of root tips. In marked contrast, d4h and dat mRNAs were associated with the laticifer and idioblast cells of leaves, stems, and flower buds. Immunocytochemical localization for TDC, D4H, and DAT proteins confirmed the differential localization of early and late stages of vindoline biosynthesis. Therefore, we concluded that the elaboration of the major leaf alkaloids involves the participation of at least two cell types and requires the intercellular translocation of a pathway intermediate. A basipetal gradient of expression in maturing leaves also was shown for all four genes by in situ RNA hybridization studies and by complementary studies with dissected leaves, suggesting that expression of the vindoline pathway occurs transiently during early leaf development. These results partially explain why attempts to produce vindoline by cell culture technology have failed.     

犁头霉11α-羟基化制备16β-甲基-11α,17α,21-三羟基孕甾-1,4-二烯-3,20-二酮

选育到一株对16β-甲基,17α,21-二羟基孕甾-1,4-二烯-3,20-二酮(Ⅱa)11α-羟基化活性强的犁头霉A28菌株,并发现底物21-乙酰化(Ⅱb)可明显提高11α-羟基化的能力.在适宜的转化条件下,Ⅱb投料浓度0.5%,产物16β-甲基-11α,11α,21-三羟基孕甾-1,4-二烯-3,20-二酮(Ⅲ)收率为73%,结构经波谱分析确认.