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1.
We have investigated the role of a novel temperature-sensitive splicing mutation, prp18. We had previously demonstrated that an accumulation of the lariat intermediate of splicing occurred at the restrictive temperature in vivo. We have now used the yeast in vitro splicing system to show that extracts from this mutant strain are heat labile for the second reaction of splicing. The heat inactivation of prp18 extracts results from loss of activity of an exchangeable component. Inactivated prp18 extracts are complemented by heat-inactivated extracts from other mutants or by fractions from wild-type extracts. In heat-inactivated prp18 extracts, 40S splicing complexes containing lariat intermediate and exon 1 can assemble. The intermediates in this 40S complex can be chased to products by complementing extracts in the presence of ATP. Both complementation of extracts and chasing of the isolated prp18 spliceosomes takes place with micrococcal nuclease-treated extracts. Furthermore, the complementation profile with fractions of wild-type extracts indicates that the splicing defect results from a mutation in a previously designated factor required for the second step of splicing. The isolation of this mutant as temperature-sensitive lethal has also facilitated cloning of the wild-type allele by complementation.  相似文献   

2.
银胶菊叶和花提取物对南方根结线虫的毒杀活性比较   总被引:1,自引:1,他引:0  
为进一步明确银胶菊(Parthenium hysterophorus L.)的杀线虫活性,对银胶菊叶和花的不同溶剂提取物、甲醇提取物的不同萃取物以及甲醇提取物碱水层的不同极性组分对南方根结线虫(Meloidogyne incognita Chitwood)的杀虫活性进行了检测,并对不同提取物、萃取物和萃取组分进行了生物碱的定性分析.结果表明:银胶菊叶和花的蒸馏水、甲醇、乙酸乙酯和石油醚提取物的得率分别为24.5%和20.3%、19.6%和10.9%、6.8%和7.7%、2.0%和2.7%,其中,叶和花的蒸馏水和甲醇提取物的杀线虫活性均较强,而石油醚提取物的杀线虫活性最弱.用质量体积分数1.0%和0.5%的叶和花蒸馏水提取物分别处理24和48 h后试虫的校正死亡率均达到100.00%;用质量体积分数1.0%和0.5%的叶和花甲醇提取物处理48 h,试虫的校正死亡率均大于90%.叶和花甲醇提取物的碱水层、三氯甲烷Ⅰ层和Ⅱ层萃取物均具有一定的杀线虫活性,其中,用质量体积分数1.0%的花和叶碱水层萃取物以及花的三氯甲烷Ⅰ层萃取物分别处理48 h,试虫的校正死亡率均为100.00%,而三氯甲烷Ⅱ层萃取物的杀线虫活性最弱.银胶菊叶和花甲醇提取物碱水层的11个不同极性组分(A1~A11)也表现出不同程度的杀线虫活性,其中,用质量体积分数0.2%和0.1%花的A2[溶剂为V(三氯甲烷)∶V(甲醇)=10∶1]和A7[溶剂为V(三氯甲烷)∶v甲醇)=1∶1]组分以及叶的A2和A6[溶剂为V(三氯甲烷)∶V(甲醇)=2∶1]组分处理48 h后,试虫的校正死亡率均达100.00%,显著高于其他组分.定性实验结果表明:银胶菊叶和花中具有杀线虫活性的提取物、萃取物和萃取组分中均含有生物碱.研究结果说明:银胶菊花的杀线虫活性高于叶片,其毒杀活性不仅与提取部位及溶剂的种类和极性有关,还与提取物浓度及作用时间等因素有关.  相似文献   

3.
The phase behaviour of total membrane lipid extracts of the blue-green alga Anacystis nidulans is compared with that of the individual lipid classes present in such extracts using fluorescence probe, differential scanning calorimetry, wide-angle X-ray diffraction and freeze-fracture techniques. Marked differences are observed in the properties of the isolated lipids as compared to the total lipid extracts. In particular, purified samples of monogalactosyldiacylglycerol and phosphatidylglycerol form complex high melting-point gel phases on storage which are not found in the membrane extracts. Addition of Mg2+ ions to the extracts is also shown to lead to an extensive phase separation of monogalactosyldiacylglycerol from the extracts. The enthalpy changes associated with phase separations occurring in the lipid extracts are found to be approx. 30% higher than those for the corresponding membranes, suggesting that the presence of other components, such as membrane proteins, may influence the phase behaviour of the lipids. The significance of these observations is discussed in terms of the factors limiting the stability of membrane systems.  相似文献   

4.
Toxic properties are more often associated with microalgal blooms than with macroalgal blooms, although herbivore deterrents are well known in macroalgae, and inhibition of invertebrate larval development by extracts from Ulva spp. has been suggested. This study was prompted from our observation that substantial discoloration of seawater occurred in small bays after mass desiccation-induced mortality of Ulvaria obscura (Kützing) Gayral. We examined the effects of extracts from Ulva fenestrata Postels et Ruprecht and Ulvaria obscura on Fucus gardneri Silva zygote development, growth of Ulva and Ulvaria , epiphytic algal accumulation, and oyster larval development. Fucus zygote development was inhibited by extracts from both species, although the effects of Ulvaria extracts were significantly greater. Epiphytic algal accumulation and the growth of Ulva and Ulvaria were inhibited by extracts from both species. Oyster larval development was arrested by the presence of extracts from each species. We conclude that extracts from both Ulva fenestrata and Ulvaria obscura have allelopathic properties. The effects are more widespread and occur at lower concentrations for extracts from Ulvaria than Ulva. These properties could alter competitive interactions by inhibiting germination or development of algae and invertebrates.  相似文献   

5.
亚硝酸盐在食材、食品加工和食品储存过程中普遍存在,它是亚硝基化合物合成的前体物质,在一定条件下亚硝酸盐极易转化为亚硝胺,二者均可致癌[1-2],因此,清除亚硝酸盐和阻断亚硝胺合成对于预防和控制癌症的发生具有重要意义。关于清除亚硝酸盐和阻断亚硝胺合成的研究已有较多报道,一些植物含有的天然活性成分,如植物中所含的多酚和黄酮类物质等,具有抗氧化、抗菌、清除亚硝酸盐和阻断亚硝胺合成的作用。顶羽菊也Acroptilon repens ( Linn.) DC.页[3]、黄花菜(Hemerocallis citrina Baroni)[4]和簕菜也Acanthopanax trifoliatus ( Linn.) Merr.页[5]等提取物具有清除亚硝酸盐和阻断亚硝胺合成的作用;黄俊生[6]从南姜也Alpinia galanga ( Linn.) Willd.页表皮中提取的色素也具有较强的清除亚硝酸钠和阻断亚硝胺合成活性。  相似文献   

6.
BackgroundExcess lipid accumulation can accelerate the development of various metabolic diseases. Blossoms of Citrus aurantium L. var. amara Engl. (CAVA) have been reported to possess inhibitory capacities on lipid deposition. However, the constituents responsible for the observed bioactivity and the underlying mechanisms are still not clearly understood.PurposeTo screen constituents from blossoms of CAVA with inhibitory effects on lipid accumulation and to explore the action mechanism.MethodsThe chloroform (CHL) extracts are prepared from blossoms of CAVA by fractional extraction and are characterized using LC-MS assay. 3T3-L1 preadipocytes are induced with differentiation medium (DMI) and treated with CHL extracts. High fat diet (HFD)-induced obese mice are further established and administrated with CHL extracts for 12 weeks. Hematoxylin and eosin (HE) staining, Oil Red O staining, ELISA, RT-qPCR, western blot and 16S rRNA gene sequence methods are employed.Results14 compounds are identified in CHL extracts and trigonelline hydrochloride, nobiletin and 7-demethylsuberosin are most abundant. CHL extracts treatment significantly inhibit differentiation of 3T3-L1 cells by regulating expression of preadipocyte factor-1 (Pref-1), fatty acid synthase (FAS) and CCAAT/enhancer binding protein α (C/EBPα). CHL extracts intervention also significantly attenuate features of obesity and improved plasma biochemical profiles in HFD-fed mice. HFD-triggered hepatic steatosis and epididymal adipose tissues (EATs) hypertrophy are also reversed by CHL extracts administration through enhancing antioxidant responses and modulating lipogenesis and energy expenditure-related genes and proteins. 16S rRNA gene sequence data further show that CHL extracts enhance the diversity of gut microbiota. CHL extracts at lower concentrations reduce the ratio of Firmicutes to Bacteroidetes and the abundance of Erysipelotrichaceae. CHL extracts at higher doses markedly increase the abundance of Lachnospiraceae.ConclusionThese findings suggest that CHL extracts probably suppress lipid accumulation through inhibiting differentiation of 3T3-L1 cells and attenuating metabolic syndromes in HFD-fed mice.  相似文献   

7.
ABSTRACT Dose-response curves are presented for the diuretic activity in aqueous extracts of brain, retrocerebral complex, and ventral nerve cord ganglia from Acheta domesticus . Diuretic activity is highest in extracts of brain and corpora cardiaca. In comparison with such extracts, those of the suboesophageal ganglia and thoracic ganglia I-III produce truncated responses, whilst abdominal ganglia 1–4 show evidence of an inhibition of the diuretic response at high doses. ED50 values, obtained from Hill plots, are similar for extracts of brain, corpora cardiaca, corpora allata, and abdominal ganglia, but are 3–4 times higher for extracts of suboesophageal and thoracic ganglia.
Separation of aqueous extracts of corpora cardiaca by reversed-phase HPLC yields a number of fractions which stimulate fluid secretion in isolated tubules. Diuretic activity in these fractions is destroyed by treatment with Pronase E, and on this basis is identified as peptidic. In general, diuretic activity is found in the same RP-HPLC fractions prepared from aqueous extracts of brain, suboesophageal ganglia, thoracic ganglia I-III, and abdominal ganglia 1–4.  相似文献   

8.
This study investigated the antioxidant and genotoxic properties of 13 South African herbal extracts. Results from the single-cell gel electrophoresis (Comet) assay indicated that there were profound differences between the plant extracts in their ability to produce DNA damage, which varied from highly genotoxic to protective. Similarly, water and methanol extracts of all the herbal preparations showed variable potencies in scavenging hydroxyl radicals, as measured by means of electron spin resonance spectrometery (ESR) with the spin trap alpha-phenyl-N-tert-butylnitrone (PBN). In general, methanol extracts were better scavengers of hydroxyl radicals than the corresponding water extracts. This was also true of the ability of these extracts to inhibit membrane lipid peroxidation, assessed with diphenyl-1-pyrenylphosphine (DPPP). However, neither methanol nor water extracts had the ability to protect against DNA damage. The results show that further research on South African traditional herbal extracts is imperative to gain understanding of the mechanisms involved in their pharmacological effects. The tests implemented in the present investigation are recommended for screening other herbal extracts.  相似文献   

9.
A newly developed cell-free system was used to study DNA repair synthesis carried out by extracts from human cell lines in vitro. Extracts from a normal human lymphoid cell line and from cell lines established from individuals with hereditary dysplastic nevus syndrome perform damage-dependent repair synthesis in plasmid DNA treated with cis- or trans-diamminedichloro-platinum(II) or irradiated with ultraviolet light. Cell extracts of xeroderma pigmentosum origin (complementation groups A, C, D, and G) are deficient in DNA repair synthesis. When damaged plasmid DNA was pretreated with purified Escherichia coli UvrABC proteins, xeroderma pigmentosum cell extracts were able to carry out DNA repair synthesis. The ability of E. coli UvrABC proteins to complement xeroderma pigmentosum cell extracts indicates that the extracts are deficient in incision, but can carry out later steps of repair. Thus the in vitro system provides results that are in agreement with the incision defect found from studies of xeroderma pigmentosum cells.  相似文献   

10.
Cytoplasmic extracts from early-passage (young), late-passage (senescent) normal human fibroblast (HF) cultures and immortalized human cell lines (HeLa, HT-1080, and MANCA) were analyzed for their ability to support semiconservative DNA synthesis in an in vitro SV40-ori DNA replication system. Unsupplemented extracts from the three permanent cell lines were demonstrated to be active in this system; whereas young HF extracts were observed to be minimally active, and no activity could be detected in the senescent HF extracts. The activity of these extracts was compared after supplementation with three recombinant human replication factors: (1) the catalytic subunit of DNA polymerase alpha (DNA pol-alpha-cat), (2) the three subunits of replication protein A (RPA), and (3) DNA topoisomerase I (Topo I). The addition of all three recombinant proteins is required for optimum activity in the young and senescent HF extracts; the order of the level of activity is: transformed > young HF > senescent HF. Young HF extracts supplemented with RPA alone are able to support significant replicative activity but not senescent extracts which require both RPA and DNA pol-alpha-cat for any detectable activity. The necessary requirement for these factors is confirmed by the failure of unsupplemented young and senescent extracts to activate MANCA extracts that have been immunodepleted of DNA pol-alpha-cat or RPA. Immunocytochemical studies revealed that RPA, DNA pol-alpha, PCNA, and topo I levels are higher in the immortal cell types used in these studies. In the HF cells, levels of DNA pol-alpha-cat and PCNA are higher (per mg protein) in the low-passage than in the senescent cells. By contrast, RPA levels, as determined by immunocytochemical or Western blot studies, were observed to be similar in both young and senescent cell nuclei. Taken together, these results indicate that the low to undetectable activity of young HF extracts in this system is due mainly to reduced intracellular levels of RPA, while the senescent HF extracts are relatively deficient in DNA polymerase alpha and probably some other essential replication factors, as well as RPA. Moreover, the retention of RPA in the senescent HF nuclei contributes to the low level of this factor in the cytoplasmic extracts from these cells.  相似文献   

11.
用不同溶剂不同方法提取美洲大蠊Periplaneta americana(L.)粪便,测定美洲大蠊各虫态的诱集活性。结果表明,采用直接浸泡提取方法,丙酮、乙醇、正己烷和二氯甲烷4种粗提物对美洲大蠊各虫态都具有明显的诱集作用,其中乙醇和丙酮粗提物的引诱效果最好,正己烷次之,二氯甲烷最弱。4种溶剂粗提物对美洲大蠊雄成虫和高龄若虫聚集活性最强,对低龄若虫聚集活性最弱。用乙醇溶剂对粪便粗提,3种提取方法均对美洲大蠊有很强的诱集效果,其中索氏抽提诱集效果最弱,直接浸泡和超声波提取效果好,且差异不显著,但直接浸泡提取效果更好。  相似文献   

12.
The herb feverfew is a folk remedy for various conditions, including inflammation, fever, psoriasis, rheumatism, and asthma. Like many herbal medicines, feverfew's mechanisms of action in the human body are largely unknown and its active ingredients remain elusive. Very often, different extraction methods of herb material produce different physical and biochemical properties and variation in clinical efficacy. We identified 3 major methods of extraction for feverfew aerial parts and used microarray technology to test the hypothesis that extracts produced by different methods elicit different gene expression profiles. We have identified approximately 200 genes that are consistently regulated by the 2 presumptive active antimigraine feverfew extracts but not associated with the inactive extract. Our results suggest that the presumptive active feverfew extracts potently stimulate more genes in human cells than the inactive extracts. We also identified several genes as unique signatures for these active extracts. All 3 feverfew extracts exhibited similar blockades on lipopolysaccharide-mediated TNF-alpha (tumor necrosis factor alpha) release, implicating that TNF-alpha is not responsible for the differences in the effects of the 3 feverfew extracts in human cells. In contrast, the active extracts more effectively suppressed CCL2 (also known as monocyte chemoattractant protein 1, MCP-1) than the inactive extracts, suggesting that CCL2 is a potential cellular target for feverfew's antimigraine effects.  相似文献   

13.
Zhang  Qishui 《Plant and Soil》1997,191(2):205-212
Effects of soil extracts from repeated plantation woodlands of Chinese-fir on soil fungi growth, the activities of microbial communities, and rates of net soil nitrogen mineralization were investigated. Soil extracts from replanted woodlands significantly inhibited soil non-pathogenic fungi growth, reduced soil respiration activities, and net soil nitrogen mineralization rates. However, soil extracts from replanted woodland increased the growth of pathogenic fungi. The combination of soil extracts and pathogenic fungi did not significantly reduce the growth of Chinese-fir seedlings when compared to the soil extracts alone. The combination of soil extracts with pathogenic and non-pathogenic fungi significantly increased the growth of Chinese-fir seedlings when compared to the combination of soil extracts and pathogenic fungi. The results suggest that the allelochemicals from soil extracts, rather than pathogenic fungi, are the key factor regulating the productivity and nitrogen cycling in repeated plantation woodlands.  相似文献   

14.
We have developed an in vitro system in which higher-order chromatin structures are assembled around naked DNAs in a cell cycle-dependent manner. Membrane-free soluble extracts specific to interphase and mitotic states were prepared from Xenopus eggs. When high molecular weight DNA is incubated with interphase extracts, fluffy chromatin-like structures are assembled. In contrast, mitotic extracts produce highly condensed chromosome-like structures. Immunofluorescence studies show that a monoclonal antibody MPM-2, which recognizes a class of mitosis-specific phosphoproteins, stains the "core" or "axis" of condensed mitotic chromatin but not interphase chromatin. By adding mitotic extracts, interphase chromatin structures are synchronously converted into the condensed state. The increasingly condensed state of chromatin correlates with the appearance and structural rearrangements of the MPM-2-stained structures. These results suggest that mitosis-specific phosphoproteins recognized by MPM-2 may be directly involved in the assembly of the chromosome scaffold-like structures and chromatin condensation. Although both extracts promote nucleosome assembly at the same rate, topoisomerase II (topo II) activity is four to five times higher in mitotic extracts compared with interphase extracts. The addition of a topo II inhibitor VM-26 into mitotic assembly mixtures disturbs the organization of the MPM-2-stained structures and affects the final stage of chromatin condensation. This in vitro system should be useful for identifying cis- and trans-acting elements responsible for higher-order chromatin assembly and its structural changes in the cell cycle.  相似文献   

15.
Watling JI  Hickman CR  Lee E  Wang K  Orrock JL 《Oecologia》2011,165(1):153-159
Water-soluble phytochemicals produced by invasive plants may represent novel elements of invaded ecosystems that can precipitate a variety of direct and indirect effects on native organisms. Phenolic compounds in particular are a common plant defense, and these compounds may have disproportionate impacts on amphibians compared to other taxa. We coupled an exploration of invasive plant extract effects on larvae of four amphibian species (the salamander Ambystoma maculatum, the toad Anaxyrus americanus, and the frogs Hyla sp. and Lithobates blairi) with behavioral observations designed to determine whether behavior can ameliorate the negative effects of exposure to invasive plant extracts. Larvae were reared in extracts of the widespread invasive Amur honeysuckle (Lonicera maackii), mixed native leaf litter, and a water control. Anaxyrus americanus tadpoles reared in L. maackii extracts were more likely to die than tadpoles reared in native extracts, but differences in mortality following rearing in native and exotic extracts were not significant for the other three species. Anaxyrus americanus and L. blairi tadpoles made more trips to the surface in L. maackii extracts than in native extracts, consistent with the hypothesis that exotic extracts may interfere with respiratory physiology and suggesting that L. blairi can behaviorally ameliorate the negative effects of L. maackii extracts. Our study highlights both a direct and indirect pathway by which invasive plant extracts may alter the ecological dynamics of native fauna.  相似文献   

16.
This report describes the purification of an endonuclease from extracts of adenovirus-type-2-infected KB cells. Endonuclease activity can also be detected in extracts of uninfected KB cells and the enzyme activities from extracts of uninfected and adenovirus-infected cells are very similar, if not identical. The enzyme has its maximal activity at pH 4.0. The enzyme found in uninfected and adenovirus-infectedcells is, however, strikingly different from an endonuclease isolated from calf serum. Hence, the endonuclease described is probably not a contaminant derived from the medium in which the KB cells were propagated. The endonuclease in crude extracts from uninfected or adenovirus-infected KB cells can be activated or its activity enhanced by treatment of the extracts with proteolytic enzymes, like pronase or trypsin. Evidence has been presented suggesting that this activation is due to proteolytic cleavage of an inhibitor present in crude extracts of uninfected and adenovirus-type-2-infected KB cells. A second endonuclease has been found in extracts of infected and uninfected cells with optimal activity at pH 7.2 and this endonuclease can be separated from the one with a pH optimum at 4.0.  相似文献   

17.
In this study, the Salmonella/microsome assay, using the micro-suspension method, was utilized to evaluate water for public supply at three sites in Rio Grande do Sul. The first site selected was in an area under industrial influence and the others were in non-industrial reference areas. Based on 40 L samples of raw water and/or after conventional treatment, compounds were extracted with XAD4 resins using natural and acidic pH, and the extracts were analyzed in the TA98 and TA100 strains with and without S9. Raw water extracts in the industrial region induced 27.4 revertants/L (rev/L) for TA100+S9 up to 226.3 rev/L for TA100-S9, both for acidic pH extracts. After conventional treatment the responses varied from 20.6 rev/L (TA98-S9) for natural pH extracts to 755.5 rev/L (TA98-S9) for acidic pH extracts. For acidic extracts obtained from reference site samples, the response, with metabolic activation only, ranged from negative to minimal. Direct mutagenic responses in acidic extracts may be elevated in treated extracts, compared with raw water extracts, influenced by the presence of by-products of the chlorination process. However, the mutagenicity observed in the treated water extracts in the industrial area increased and reflected mainly the combination of directly and indirectly acting compounds in the source waters, that are heavily influenced by anthropogenic factors.  相似文献   

18.
Glycolipids that bind 125I-labeled lectins are detected by autoradiography after thin layer chromatography of glycolipid standards or crude lipid extracts. Soybean agglutinin, Bandeiraea simplicifolia I isolectins A4 and B4, and Helix pomatia lectin are used to detect corresponding cell surface, glycolipid receptors in human and bovine erythrocytes. When lipid extracts from A and AB erythrocyte stroma are analyzed with Helix pomatia lectin, a polymorphic expression of blood group A glycolipid determinants is detected. The Bandeiraea simplicifolia isolectins react weakly with human erythrocyte glycolipids but bind at least 4 glycolipids in bovine stroma extracts. Soybean agglutinin reacts with glycolipids in all erythrocytes analyzed. This technique extends lectin specificity studies from inhibition analyses in aqueous systems using available, known structures to identification of specific, lectin-binding glycolipids in crude lipid extracts of cell membranes.  相似文献   

19.
J. Loch 《Plant and Soil》1985,83(1):77-83
Summary On the basis of pot experiments carried out on soils with different Mg contents using ryegrass, it could be established that Mg uptake by ryegrass is closely related to Mg in the EUF extracts. The Mg contents in the EUF extracts are therefore suitable indices of actually and potentially available Mg in soils.The ratio between Mg in the EUF extracts and Mg deposited on the electrode + filter varies. The absolute amount of Mg in the EUF extracts increases with increasing Mg content of the soils. The higher, however, the total desorbable Mg the lower are the Mg contents in the EUF extracts in comparison with the Mg contents deposited on electrode + filter. Besides total desorbable Mg the EUF-Mg fractions are influenced as well by the organic and inorganic CEC and by soil pH. The amount of EUF filtrate flowing through filter and electrode also has an influence on the amount of Mg in the EUF filtrate.Consequently for a proper evaluation of available soil Mg total EUF-Mg (Mg in the EUF filtrate + Mg deposited on electrode + filter) should be used.  相似文献   

20.
Ontogeny of immunoreactive CCK and VIP in pig brain and gut   总被引:1,自引:0,他引:1  
The concentrations and hormonal forms of CCK and VIP have been determined in extracts of the brain and duodenum of the developing and adult pig. In methanol extracts of the brain cortex, the single hormone form, CCK8, increased from 130 +/- 20 (Mean +/- SEM) pmol/g at birth to an adult level of 300 +/- 50 pmol/g. In acid extracts of brain, the predominant immunoreactive form had N-terminal immunoreactivity and increased from 240 +/- 20 pmol/g at birth to an adult level 490 +/- 30 pmol/g; the C-terminal immunoreactivity was about 10-fold lower. The concentrations and hormonal forms of immunoreactive CCK in duodenal extracts did not appear to be age-related. C-terminal immunoreactivity in methanol extracts averaged 140 +/- 20 pmol/g and in acid extracts 240 +/- 60 pmol/g. The concentration of N-terminal immunoreactivity in acid extracts averaged 490 +/- 70 pmol/g. The VIP concentrations in acid extracts of the brain cortex was 13.5 +/- 2 pmol/g at birth and rose gradually to 30 +/- 9 pmol/g in the adult; in duodenal extracts it was 240 +/- 18 pmol/g at birth and 195 +/- 38 pmol/g in the adult. These results are in marked contrast with the ontogeny of these hormones in the rat in which brain concentrations of CCK and VIP in the neonate are less than 10% of adult levels and in which there are age-related changes in the content of these hormones in the duodenum as well.  相似文献   

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