灵芝及灵芝孢子提取物的抗肿瘤及对DNA拓扑异构酶作用的研究

目的:观察灵芝子实体水提物及醇提物混合品(灵芝提取物)、灵芝孢子粉提取物(油)的体内外抗肿瘤作用,并测定它们对DNA拓扑异构酶Ⅰ、Ⅱ的作用。方法:台盼蓝计数法、MTT法观察体外对肿瘤细胞的抑制作用;应用小鼠移植瘤模型研究体内抗肿瘤作用,同时测定荷瘤小鼠的胸腺指数及脾指数;DNA超螺旋解旋法测定拓扑异构酶Ⅰ、Ⅱ活性。结果:灵芝提取物对K562、HL60、SGC-7901细胞作用24h的IC50值分别为0.44mg/ml、0.39mg/ml、0.90mg/ml,灵芝孢子油对K562、HL60、SGC-7901细胞作用24h的IC50值分别为1.13mg/ml、2.27mg/ml、6.29mg/ml;体内实验中,对S180、H22移植瘤株的抑制率灵芝提取物4g/kg分别为39.1%、44.6%,灵芝孢子油1.2g/kg分别为30.9%、44.9%;灵芝提取物1g/kg和2g/kg对S180的抑制率分别为15.6%和19.0%,对H22的抑制率分别为28.8%和38.5%;灵芝提取物对荷瘤小鼠的免疫功能有一定的上调影响,灵芝孢子油则无明显作用;两者对拓扑异构酶Ⅰ、Ⅱ的活性均有抑制作用。结论:灵芝提取物、灵芝孢子油均具有体内外抗肿瘤作用,二者的抗癌机制与抑制拓扑异构酶Ⅰ、Ⅱ的活性有关。

怎样成为造物主—《孢子》（Spore）告诉你的故事

威尔·怀特，是与彼得·莫里纽斯（前牛蛙工作室，现狮子头工作室主创）、罗伯特·威廉姆斯（《国王秘史》系列制作人）并称为“游戏之神”的天才制作人。提到威尔·怀特，自然想到的便是其制作的模拟系列。

暗黑孢子吹响反击暗黑军团的号角……

大凡玩过由业内知名的游戏公司Maxis于2008年9月推出的《孢子》（Spore）的玩家相信都会认同笔者以下的两个观点。

Comparative Studies on the Changes of Microtubule Distribution and Reorganization During the Meiotic Stages of Development in Normal (IR36) and a Temperature/photoperiod Sensitive Male Sterile Line (Peiai 64S) of Rice ( Oryza sativa )

Changes in the pattern of organization of microtubules in the meiotic stages of development of pollen (i.e. from pre-meiotic interphase to more or less metaphase I) of a normal (IR36) and a temperature/photoperiod sensitive male sterile line (Peiai 64S) of rice were studied using immunofluorescence confocal microscopy. In IR36, from pre-meiotic interphase to metaphase I, the pattern of microtubule distribution in the meiocytes underwent a series of changes. Some new organizational patterns of microtubules (that have not been described before) were observed during microsporogenesis, including the existence of a broad band of perinuclear microtubules at the diakinesis stage of development. The pattern of microtubule distribution in the meiocytes of the male sterile line, Peiai 64S, was quite different front that seen in IR36. In Peiai 64S, the microtubules showed abnormal patterns of distribution from pre-meiotic interphase to metaphase I. For example the broad band of perinuclear microtubules seen at diakinesis in IR36 was much disorganized and loosened in Peiai 64S. The spindles formed were also very abnormal and different from the normal spindle. The appearance of abnormal microtubule distribution in the early stages of microsporogenesis may contribute to the malformation and ultimate abortion of pollen in Peiai 64S.

Embryological Studies on Facultative Apomixis in Apluda mutica (Gramineae)

A cytological study on Apluda mutica L. revealed that this species was a facultative apomict with 60.74% of apomictic embryo sac formation. Its sexual reproduction exhibited the characteristics of polygonum_type embryo sac formation. After the 3 megaspores at the micropylar end had degenerated, leaving only the chalazal megaspore in the nucellus, a nucellar cell adjacent to the megaspore differentiated into an aposporous initial, which later developed into a panicum_type embryo sac. It was an aposporous 4_nucleate embryo sac without chromosome reduction and composed of an egg, a single synergid and a central cell with 2 polar nuclei. The chalazal megaspore degenerated before the onset of the division in the aposporous initial cell.

Microtubule Structure and Male Sterility in a Gene-Cytoplasmic Male Sterile Line of Rice, Zhen Shan 97A

Histological changes that occur during microsporogenesis are documented in a gene-cytoplasmic male sterile rice ( Oryza saliva L.) line, Zhen Shan 97A, its maintainer line, Zhen Shan 97B, and the restorer line, Ce64 of a Mine hybrid rice production system. In the restorer line, Ce64, the developing microsporocytes have dense cytoplasm and a distinct set of circumferential microtubules around the nucleus. Successive cytokinesis results in the formation of tetrads. The microtubules within the cells of tetrads and microspores radiate from the surface of the nucleus towards the outer edge of the cytoplasm. Subsequent pollen development is normal. During the course of microspore formation tubulin speckles can be found in the cytoplasm. The general pattern of development and microtubule organization in the maintainer lined Zhen Shan 97B, is similar to Ce64, except that a few more tubulin speckles appear during microspore formation. In the case of the mate sterile line, Zhen Shan 97A, a number of abnormalities can be discerned during early microsporogenesis. These include vacuoles forming within the developing microsporocyte and faintly stained microtubules with no defined distribution pattern. Prominent tubulin speckles are common within the cytoplasm. For those microsporocytes that undergo meiosis, no defined organizational patterns of microtubules can be found within the tetrad. All microspores abort soon after. Abnormalities and defects in microtubule organization observed in Zhen Shan 97A showed that complex interactions between the cytoplasm and the nucleus began at very early stage of microsporocyte development.

Studies on the Triterpenoid Constituents of the Spores from Ganoderma lucidum Karst

Five compounds were isolated from the ether-soluble fraction of the spores of Gano- derma lucidum.On the basis of their chemical properties and spectral data(MS,UV,IR,~1H and ^(13)CNMR),they were identified as 3,7,11,12,15,23-hexaoxo-5α-lanosta-8-en-26-oic acid(Ⅰ),gano- deric acid B(Ⅱ),C(Ⅲ),D(Ⅳ)and ganodermanontriol(Ⅴ).Compound Ⅰ is a new natural product, named ganosporeric acid A.Compounds Ⅱ,Ⅲ,Ⅳ and Ⅴ are known compounds and were obtained for the first time from the spores of Ganoderma lucidum.Pharmacological experiments showed that ganosporeric acid A has an activity for lowering the levels GPT in mice with liver injury by CCl_4 and GaNI and exhibits heptoprotective effects.

Growth and Development of Staminate Inflorescence and Anatomic Observation of Male Chestnut Flower

The growth and development of staminate inflorescence and anatomic structure of male chestnut flower were observed. Results showed that staminate in-florescence on the base of branch formed first, then upward successively. About 50 days were needed from the formation of staminate inflorescence on the base of branch to ful y develop the staminate inflorescence on the top of the branch. On the same staminate inflorescence, male flower clusters of the base formed first, then upward successively. About 20 days were needed from the formation of stami-nate inflorescence on the base of the male flower cluster to ful y develop the stami-nate inflorescence on the top of the branch. 5-7 male flowers forming a cluster, the flower number in a cluster was odd number usual y, and there was one on the top and each two paral el y arranged downward. The flower on the top came into bloom first, and then downward successively. The flowers paral el y arranged came into bloom at the same time. Sporangium of male flower of chestnut was monolocular. There were a large number of pol en grains in the sporangium. There were large differences between the development process of different sporangium in one male flower. Chestnut had larger quantity of male flowers and pol en and long period of pol ination compared with female flower. It is remained to be further studied whether it was necessary for anemophilous pol ination.

Antitumor Activity of the Ganoderma Lucidum Spore Alcohol Extract in Vitro

Several cancer cell lines(epithelioma cells or leukemia cells)from human being or mouse were first used to study the antitumor activity of the Ganoderma lucidum spore alcohol extract(GLSAE)in vitro by the MTT test A comparision was made between the sporodermbroken(SB)and sporoderm nonbroken(SN)GLSAE It was showed that both GLSAE SB and GLSAE SN could inhibit the proliferation of these cancer cells,but the activity of GLSAE SB was much higher than that of GLSAE SN These results suggested that Ganoderma lucidum spore could probably be used for tumor treatment

Inhibition of Chlamydospore Germination and Mycelial Growth of Trichoderma spp. by Chemical Fungicides

For exploring the influences of application and residue of chemical fungi- cides on chlamydospore preparations of Trichoderma spp., the effects of seven chemical fungicides on chlamydospore germination and mycelia growth of two bio- control fungi T. harzianum 610 and T. Iongibrachiatum 758 were studied. Carben- dazim, tebuconazole and difenoconazole showed strong toxicities, thiram and car- bexin showed moderate toxicities, and metalaxyl showed Mycelia growth of the two strains was more sensitive to most tested fungicides than those of chlamydospore germination. Chlamydospore germination of 610 was more sensitive to tested fungicides than those of 758, and mycelia growth of 758 was more sensitive to most tested fungi- cides than those of 610. Among the seven fungicides, 98% carbendazim had the strongest effects （ECru values were 1.64 and 0.05μpg/ml）, and 70% pentachloroni- trobenzene had the weakest effects （EC50 values were 1.64 and 0,05 μg/ml） for chlamydospore germination and mycelia growth of 610. As for 758, 98% carbendaz- im had the strongest inhibitory effects and 95% metalaxyl had the weakest inhibitory for chlamydospore germination of 756 （EC50 values were 0.62 and 1 108.61 μg/ml respectively）, whereas 96.2% tebuconazole showed the strongest inhibitory effects for mycelia growth of 758 （EC= value was 0.32μg/ml）, and 95% metalaxyl was the weakest （EC= value was 1 206.29 μg/ml）. According to the applied concentration of different fungicides in practice, we concluded that chlamydospore preparations of 610 and 758 could not be combined with carbendazim, tebuconazole, thiram and carboxin for controlling plant diseases, and the pesticide residues to the biocontrol effects should be kept in mind. Chlamydospore preparations of 610 and 758 can be and difenoconazole for controlling plant dis- eases, 758 chlamydospore preparations and germinated chlamydospore of 610 can be combined with metalaxyl for controlling plant diseases, and pesticide residue risk was not serious.

Facultative Apomixis in an Endangered Dioecious Species, Woonyoungia septentrionalis (Magnoliaceae)

The breeding system of an endangered dioecious species, Woonyoungia septentrionalis (Dandy) Law, was studied in two natural populations, the Mulun population and the Dahuangni population, in Guangxi Province, China from June 2001 to early September 2002. The female flowers from previously bagged buds were treated respectively with paper bags, net bags and hand pollination by comparing with natural pollination. The results showed that female flowers treated with paper bags, net bags, hand pollination, and natural pollination all set fruits and seeds, but there was great difference in their fruit set, seed set, pre-emergent reproductive success (PERS) and seed germination rate for two populations. In both populations, PERS of hand pollination and natural pollination were higher than those of paper bagged and net bagged treatments. Among them, PERS of hand pollination was the highest, and PERS of net bagged treatment was the lowest. Both sexual reproduction and apomixis occurred simultaneously in W septentrionalis and seeds from both sexual and apomixis could germinate into seedlings. Therefore, the breeding system of W septentrionalis belonged to facultative apomixis. The apomixis in Magnoliaceae was reported for the first time.

A Discovery of Anomozamites with Reproductive Organs

Anomozamites haifanggouensis (Kimura et al.) Zheng et Zhang comb. nov. is a characteristic Bennettitalean plant in Middle Jurassic flora. Only one specimen was collected from Haifanggou Formation (Middle Jurassic) in Daohugou village, Shantou town, Ningcheng County, Nei Mongol Autonomous Region, China. The stem of plant is slender and repeatedly branches as a dichasial system with a fertile shoot in the fork, forming equal and widely divergent branches, on the branches born with the fronds of Anomozamites type. Its reproductive organs consists of some microsporophylls and a few of the bracteoid small leaves (or receptacular leaves). Some scattered bracteoid leaves and microsporophylls were formerly considered by Pan as dicots. Since then, they were proposed to be a non-committal genus Pankuangia and described them as P. haifanggouensis by Kimura et al. In this study, we discover that these bracteoid small leaves and microsporophylls should be of Anomozamites. This discovery provides strong evidence for the 'Pankuangia' determination of the taxonomical position.

Primary study on Nosema bombycis Infecting the Insect of Drosophila

[ Objective ] The aim of this study was to investigate the infectivity of Nosema bombycis to drosophila, which offered a new vision for systematical studies on the infection mechanism of Nosema bombycis, and also provided reference for the bio-control effect of Nosema bombycis. [ Method ] Nosema bombycis was used to feed wild type and mutant drosophila, and the morphological observation of Nosema bombycis in drosophila body fluid was also analyzed by calcofluor white M2R fluorescent staining. [ Result] Nosema bombycis could infect drosophila, and the number of Nosema bombycis in the infected mutant drosophila was higher than that in wild type drosophila. [ Conclusion ] Nosema bombycis can infect drosophila, which provides primary reference for studies on the infectivity of Nosema bombycis to other hosts and also lays a foundation for further study on the infection mechanism of Nosema bombycis.

Middle Jurassic Sporo-pollen Assemblages in Northern Turpan Depression, Xinjiang, Northwest China

Eighty-one species of fossil spores and pollen assigned to 52 genera are found from the Middle Jurassic Xishanyao, Sanjianfang and Qiketai Formations of boreholes Le 1, Mi 1, Shan 1 and Dun 1 in the northern Turpan Depression, Xinjiang, northwest China. Three assemblages have been proposed as: Cyathidites-Psophosphaera-Cycadopites assemblage, Cyathidites-Quadraceculina-Classopollis assemblage and Cyathidites-Callialasporites-Classopollis assemblage obtained from the Xishanyao Formation, the Sanjianfang Formation and Qiketai Formation respectively. Based on the sporo-pollen assemblage features and con-elation, their geological ages are regarded as of Middle Jurassic, probably Aalenian to Bathonian. The results of the study indicate that the vegetation of early Middle Jurassic in this region consisted mainly of Gymnospermae of Ginkgoaceae of Cycadales, Podocarpaceae, Pinaceae and Pteridophyta of Cyatheacene, Osmundaceae, Lycopodiaceae and Dicksoniaceae, etc. They reflect a warm-moist subtropic climate. To the middle-late Middle Jurassic, Cheirolepidaceae was dominant in the vegetation, which indicates that the climate was in the trend of becoming dry.

Antifungal Effect of Streptomyces 702 Antifungal Monomer Component DZP8 on Rhizoctonia solani and Magnaporthe grisea

The aim of this study was to investigate the in vitro antifungal effects of antifungal monomer component DZP8 isolated from Streptomyces 702 on the mycelium growth, sclerotium formation and germination of Rhizoctonia solani and on the mycelium growth, conidial formation, germination, appressorium formation of Magnaporthe grisea. The results showed that the antifungal monomer component DZP8 has strong antifungal effect on both the R. solani and M. grisea. The EC50 and EC90 of DZP8 were 1.81 and 3.35 μg/ml on Ft. solani respectively, and 37.01 and 136.21 μg/ml on M. grisea respectively. Under the treatment of 48.01 μg/ml DZP8, the sclerotium formation rate of R. solani was just 39.21%, the formation time delayed by 216 h and the dry weight decreased by 81.37% in comparison the con- trol; and 33.51 μg/ml DZP8 significantly inhibited the sclerotium germination. In the presence of 160.08 μg/ml DZP8, the sporulation of M. grisea was just 9.29% of control sample; 20.14 μg/ml DZP8 inhibited the conidial germination suppression rate by 95.16%, and the appressorium formation by 100%.

Studies on Enhancing Yield of Embryos in Microspore Culture of Brassica napus L.

In ecological zone of Chengdu, Sichuan, microspore culture was carried out in Brassica napus L. to study the influencing factors on microspore culture. The results showed that the temperature on microspore formation stage, day and night temperature, disinfection solution of buds, cultivation concentration on microspore and strain-age were both important influencing factors on microspore culture. At a temperature below 5 ℃ or above 20 ℃, the material had a much lower embryo producing rate or even could not produce any embryo, but at the optimum temperature of 10 -15 ℃ the embryo yield was up to 300 pieces per bud; the best cultivation effect appeared when 0. 1% HgCl2 was used for disin- fection; the best density of microspore was 3 -4 buds per dish; In 2009, while strain-age was from 125 d to 150 d, the microspore embryo yield increased as strain-age increased. When stain-age was 150 days, the microspore embryo yield was up to the highest, but the yield declined after 150 days.

Embryogenesis and Development of Isolated Microspore in Chinese Cabbage

[Objective] The aim was to observe embryogenesis and development of isolated microspores in Chinese cabbage. [Method] Chinese cabbage F 1 hybrids were used as the experimental materials, and optical microspore was employed to observe the embryogenesis and development of isolated microspores. [Result] Cells swelled after heat shock treatment, which was the critical factor of embryoid induction. Three pathways equal division, unequal division and germination of microspores were discovered to lead to the embryogenesis from isolated microspores after swelling. Microspore could grow directly to embryoid through germination path way. Equally divided microspores formed the original embryos after successive multiple equal divisions. Original embryos could develop into cotyledon-shaped embryos via globular, heart-shaped and torpedo-shaped embryos. The large one of the two cells from unequally divided microspores continued to divide and finally formed a polar embryoid. [Conclusion] The study will provide cytological basis for high induction frequency and embryoid of Chinese cabbage.

Establishment on Isolated Microspore Culture Optimized System for Restorer of New Cytoplasmic Male Sterile (NER) of Brassica napus L.

[Objective]The aim was to establish optimized system for NER isolated microspore culture of Brassica napus L.. [Method]Twenty varieties of NER of Brassica napus were grown under uncontrolled temperature and light conditions,and their isolated microspore from anthers were used as explants in vitro culture. The influencing factors of microspore culture were preliminarily studied. [Result]The difference of genotypes was important influencing factors to embryoid yield. The embryoid yield increased by supplementing with 6-BA and NAA,culturing in solid-liquid double layer medium with activated charcoal; The difference was not significant of embryoid yield between culturing in medium supplemented with colchicines and the CK. The rates of cotyledonous embryoids directly developed into normal plantlets increased through enriching with 0.1-0.2 mg/L NAA and being treated on slim illumination two days before being inducted into normal plantlets. [Conclusion]The technical system of microspore culture of restorer of new cytoplasmic male sterile （NER） was established,which and lays a foundation for accelerating genotype purification of NER introgressive line.

Construction of Eukaryotic Expression Vector with Partial Encoding Sequence of Actin from Cryptosporidium andersoni

[Objective] To clone the actin gene of Cryptosporidium andersoni, and to study its eukaryotic expression in Hela cells. [Methed] Specific primers were designed for the partial encoding sequence of actin, which were obtained by screening the T7 phage display library of Cryptosporidium andersoni, and the actin gene CA42 was amplified by PCR. Recombinant eukaryotic expression plasmid pVAX1-CA42 was constructed and transfected to Hela cells with lipofection strategy. Indirect im- munofluorescence staining, SDS-PAGE and Western blotting analysis were used to detect the expression of recombinant protein in Hela cells. [Result] CA42 protein was successfully expressed in Hela cells, and the expression products had reactogenicity. [Conclusion] The partial encoding sequence of actin from Cryptosporidium andersoni has been successfully cloned, and it can be stably expressed in Hela Cells

Investigation of Biological Characteristics of Parasitic Fusarium Fungi from Tibetan Cordyceps sinensis

[Objective] This study aimed to investigate the biological characteristics of parasitic Fusarium fungi from tibetan Cordyceps sinensis. [Method] A Fusarium strain was isolated from fresh tibetan Cordyceps sinensis as the experimental material, to observe the morphologies of Fusarium mycelium and spores. [Result] The isolated Fusarium fungus strain grew lushly on rice medium, with white and villiform myceliurn, and the villous stroma appeared in colors from beige to pink. After cultivated at （18 ± 4） ℃ for 14 d, the Fusarium strain produced milky white protrusions containing pale yellow chlamydospores, which can be divided into microconidiums and macroconidiums based on the different sizes. According to Ainsworth＇s classification system, the isolated strain belongs to Fusarium, Tuberculariales, Hyphomycetes, adelomycete subphylum. Further observation identified three types of macroconidium, and two of them showed distinct morphology, which was not described in Ainsworth＇s classification system. [Conclusion] The structure of the strain isolated from tibetan Cordyceps sinensis is significantly different from that of Fusariurn fungi recorded in relevant literatures, which requires further investigation.