首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 296 毫秒
1.
为了探讨花斑裸鲤(Gymnocypris eckloni)血红蛋白时序转换 利用花斑裸鲤全基因组数据鉴定胚胎/仔鱼型血红蛋白基因家族成员, 并通过整胚原位杂交方法, 检测花斑裸鲤胚胎/仔鱼型血红蛋白基因在胚胎发育不同阶段的表达及定位。结果表明, 花斑裸鲤基因组中共鉴定到5个胚胎/仔鱼型血红蛋白基因, 分别为hbae1、hbae4、hbae5、hbbe1和hbbe3, 与斑马鱼(Danio rerio)相比, 花斑裸鲤基因组缺少hbae3和 hbbe2 基因, 暗示第四轮全基因组复制事件后所经历的小规模基因删除事件在花斑裸鲤特异性血红蛋白基因形成中发挥了重要作用。整胚原位杂交结果显示, hbae1基因在胚胎发育的120h至432h内持续表达, hbbe1基因在96h开始表达持续至432h, hbbe3基因杂交信号出现在胚胎发育120h至384h内, 在胚胎发育全过程中未能观察到hbae4和hbae5基因的杂交信号。杂交信号主要位于胚胎正中轴、后部侧向中胚层、背主动脉腹侧区、尾部造血区及卵黄。正义探针作为阴性对照, 在胚胎发育阶段均无任何杂交信号。花斑裸鲤具有与其他鱼类不同的胚胎/仔鱼型血红蛋白基因家族成员及血红蛋白转换表达特征; hbae1、hbbe1和hbbe3基因在花斑裸鲤早期胚胎发育过程中发挥重要作用, 而hbae4和hbae5基因的生物学功能可能有所弱化。  相似文献   

2.
为阐明拟赤梢鱼(Pseudaspius leptocephalus)胚胎发育和仔稚鱼发育特点, 采用人工催产的方式获得受精卵, 观察分析了拟赤梢鱼胚胎发育和仔稚发育的时序特征。结果表明: 拟赤梢鱼成熟卵粒为黄色圆球形, 平均卵径为(1.77±0.20) mm, 遇水具微黏性; 在水温23℃条件下, 胚胎发育经历合子期、卵裂期、囊胚期、原肠胚期、神经胚期、器官形成期和孵化出膜期7个阶段26个时期, 共历时47h 55min完成孵化过程。初孵仔鱼在(23±1)℃水温条件下, 经历卵黄囊期仔鱼(0—7d)、晚期仔鱼(8—26d)和稚鱼期(27—31d), 进入幼鱼期; 卵黄囊期仔鱼游泳能力差, 随着卵黄囊逐渐消耗, 体表色素斑、胸鳍和尾鳍等逐渐形成, 消化道贯通, 鳔充气; 晚期仔鱼卵黄囊完全消失, 仔鱼游泳能力增强, 开口摄食, 腹鳍形成, 皮肤透明; 稚鱼期鳞片形成并覆盖全身, 鱼体形态已逐渐与成鱼无异。拟赤梢鱼仔稚鱼阶段全长生长特性公式为TL=0.0125x2+0.3579x+6.2058 (R2=0.9953), 出膜15d内, 仔鱼生长速度缓慢, 全长日生长率仅为(0.38±0.02) mm/d, 15d后, 仔鱼生长速度变快, 全长日生长率可达(1.24±0.09) mm/d。研究初步阐明了拟赤梢鱼的胚胎发育和仔稚鱼发育的时序特征, 为拟赤梢鱼的苗种规模化繁育提供了理论基础。  相似文献   

3.
白斑狗鱼胚胎和仔鱼发育的研究   总被引:20,自引:0,他引:20  
白斑狗鱼 (EscoxluciusLinnaeus) ,卵圆球形 ,呈金黄色 ,直径为 1.9— 2 .2mm ,具黏性。在 5— 11℃的温度下 ,受精卵历时 185h 18min发育后孵出 ,所需总积温为 2 6 4 1 37℃ ,胚胎发育可划分为 6个阶段 ,33个发育时期。初孵仔鱼全长 7 0— 7 7mm ,从孵出到各鳍形成需 2 1d。胚胎发育各期及仔鱼的主要形态特征均被描述  相似文献   

4.
兔植入前移核胚中发育相关基因的差异表达分析   总被引:10,自引:0,他引:10  
与早期胚胎发育相关的一些重要基因异常表达致使克隆胚细胞核的再程序化过程受阻,是导致动物克隆失败的重要原因.为了分离鉴定再程序化相关基因,我们改进了mRNA差异显示技术,成功地建立了单胚差示技术体系.以不同发育时期的兔克隆胚(MⅡ卵、2细胞、4细胞、8~16细胞克隆胚胎)为材料进行单胚差示, 分离了80个差异片段.经反向RNA印迹验证、亚克隆、序列分析及NCBI GenBank数据库检索, 结果表明:A028片段与CstF3基因有93%的同源性, 在早期胚胎发育过程中的表达有阶段特异性, 该基因在兔克隆胚的早期发育过程中起重要作用.RNA印迹显示:该基因在所检测的组织中,只在卵巢中有表达.这项研究为再程序化相关基因全长的克隆及功能研究奠定了良好的基础.  相似文献   

5.
研究对人工繁殖的北极茴鱼(Thymallus arcticus grubei)胚胎发育开展系统观察,记录分析其胚胎及仔鱼发育各时期的形态特征,旨在为北极茴鱼的人工繁育和种质资源保护提供必要的基础数据。结果显示,北极茴鱼受精卵呈圆球形,金黄色,沉性卵,未吸水卵径(2.46±0.14) mm,吸水卵径(3.14±0.18) mm,卵黄质内有多个油球。油球的数量和空间分布在胚胎发育过程中发生了规律性变化。在孵化水温(11.06±0.72)℃,溶氧8.3—9.8 mg/L条件下,历时301h完成整个胚胎发育过程,所需积温为3384.84h·℃,经历合子期、卵裂期、囊胚期、原肠胚期、神经胚期、器官形成期和孵化出膜7个阶段,共26个时期。北极茴鱼仔鱼尾鳍、胸鳍的分化和眼色素沉积在受精卵胚胎发育后期就已经完成,背鳍、腹鳍、臀鳍和脂鳍等在胚后发育过程中相继分化。其初孵仔鱼平均全长为(9.33±0.35) mm,仔鱼卵黄囊呈圆球形, 18日龄时卵黄囊和油球被完全消耗。其早期发育阶段(0—16日龄)的生长特性符合公式:y=0.0005x4–0.0201x3+0....  相似文献   

6.
通过Bouin's液、5%的福尔马林、透明液固定和活体解剖观察等4种不同方法,对白点鲑(Salvelinus leucomaenis)胚胎和仔鱼发育进行了系统观察,描述了早期发育过程。白点鲑受精卵为端黄卵,沉性,橙黄色,呈圆球形。在水温3.40~8.89℃,受精卵历时1 944 h,经历6个阶段的胚胎发育破膜孵出仔鱼;初孵仔鱼全长(17.89±0.32)mm,破膜后73 d各鳍条发育完全,并出现"幼鲑斑",破膜后350d鱼体外部形态与成鱼基本相同。将白点鲑与几种鱼类进行了对比,并且探讨了其胚胎发育特点。经比较4种不同观察方法,Bouin's液固定后剥离卵膜观察是研究白点鲑等卵膜较厚鱼类的理想方法。  相似文献   

7.
克隆参与胚胎发育的新基因并研究其表达规律和功能是揭示胚胎发育的基因调控机理的重要途径。囊胚形成和原肠形成是哺乳动物胚胎发育过程中的两个关键阶段。囊胚阶段发生了胚胎的第一次分化,是细胞多能性和分化的一个转折点。此时涉及的基因活动,既有维持胚胎干细胞全能性或多能性的基因活动,又有按照预定发育模式参与胚胎定向分化的基因活动。原肠期是胚胎发育过程中的第二个关键转折点,涉及到3个胚层的形成和细胞命运决定等多种变化。在这个时期胚胎获得了胎儿原基的所有信息,新组织的产生和细胞迁移的再生组织与形态发生、细胞增殖、细胞分化、模式形成等存在着非常复杂而相互协调的关联。大多数细胞正由原来的多潜能逐渐向寡潜能发展,控制组织器官形态建成的基因正逐渐开启。这两个时期的基因表达图式、特征和种类会有很大的差异和变化,因此研究这两个时期的新基因的表达规律和功能,将是了解胚胎发育的基因调控机理的重要途径。文章以这两个时期胚胎为原始材料,利用减法杂交方法克隆到一新的小鼠胚胎基因mED2,对其进行了表达规律和生物学功能的初步分析。RT-PCR-Southern和原位杂交实验表明,mED2基因转录水平具有发育阶段的依赖性;随着发育过程的进行,其表达主要在胚神经系统和中胚层衍生的组织表达。mED2基因活性的knockdown对于合子的卵裂和植入前早期胚胎发育均有抑制作用。亚细胞定位实验表明,mED2基因编码的蛋白基本定位于细胞核膜及其临近的内膜细胞器(粗糙内质网和高尔基体)。根据生物信息学分析,mED2蛋白可能为一跨膜蛋白且与含有硫氧还蛋白结构域的蛋白有部分匹配。由此推测mED2基因参与了小鼠植入前早期胚胎发育,其基因产物可能通过蛋白之间的相互作用,即对蛋白进行后期修饰、折叠及行使分子伴侣等作用来活化或抑制其靶蛋白的活性,进而参与小鼠的早期胚胎发育。  相似文献   

8.
青鱼胚胎发育中乳酸脱氢酶(LDH)同工酶的研究   总被引:4,自引:0,他引:4  
本文报道了LDH同工酶基因在青鱼胚胎发育过程中表达的时序。研究结果表明,LDH-B基因在整过发育过程中部处于活动状态;胚胎发育早期的LDH同工酶谱与成熟卵的酶谱相同;LDH-A_2B_2、-A_3B和-A_4三种同工酶在胚胎发育至出膜前期后才出现。  相似文献   

9.
作者对分布于北京市怀柔区怀九河的宽鳍鱲(Zacco platypus)的胚胎发育及仔鱼前期发育进行了研究,过程中采用人工干法授精获取受精卵, 观察并描述了宽鳍鱲早期发育过程及其特点。结果显示, 宽鳍鱲成熟卵呈圆球状, 平均卵径1.04 mm, 为沉性卵。在平均23.0℃(17.1-28.0℃)水温条件下, 从受精卵到孵化经历了73h1min, 积温为1682.3 度时; 孵化后6.5d 进入弯曲期仔鱼。仔鱼前期发育速度与出膜前相比明显减慢;弯曲期仔鱼出现大量死亡可能与有限的人工培育条件、混合营养期能量供给不足等原因有关。通过比较发现, 宽鳍鱲与鲤科中其他21 个种相比, 早期发育时间比其中7 个种均长。宽鳍鱲早期发育时间比同域分布、相同发育水温的马口鱼略长, 明显长于同属的纵纹鱲的发育时间。宽鳍鱲南、北方种群仔鱼发育速度存在差异, 北方种群出膜前发育速度比南方种群快, 但出膜后发育速度减慢。    相似文献   

10.
以甘蓝型油菜(Brassica napus L.)品种‘Westar’和‘Topas’为材料,通过超微结构观察和荧光定量PCR技术对油菜胚胎发育早期油体的发生、油体蛋白及脂肪酸合成转录因子基因的表达情况进行分析。结果显示:油体出现在油菜胚胎发育早期,在授粉9~11 d后(球形胚时期)的胚体和胚柄中均存在直径小于0.5 μm的油体;荧光定量实验结果表明,除BnCLO3的表达量在整个胚胎发育阶段无明显变化外,其他油体蛋白基因Oleosins、Steroleosins和BnCLO1的表达量在心形胚时期就明显增多并持续增长;脂肪酸合成转录因子BnLEC1、BnL1L、BnWRI1和BnFUS3在胚胎发育阶段,基因表达规律均呈先上升再下降的趋势,但达到最高值的时间存在差异,其中BnLEC1最早,BnL1L其次,BnWRI1和BnFUS3较晚。研究结果表明甘蓝型油菜在球形胚时期出现油体,其结构蛋白和转录调控因子基因的表达自心形胚开始明显增多。  相似文献   

11.
Teleosts show a great variety in visual opsin complement, due to both gene duplication and gene loss. The repertoire ranges from one subfamily of visual opsins (scotopic vision) including rod opsin only retinas seen in many deep-sea species to multiple subfamilies of visual opsins in some pelagic species. We have investigated the opsin repertoire of Atlantic cod (Gadus morhua) using information in the recently sequenced cod genome and found that despite cod not being a deep sea species it lacks visual subfamilies sensitive towards the most extreme parts of the light spectra representing UV and red light. Furthermore, we find that Atlantic cod has duplicated paralogs of both blue-sensitive SWS2 and green-sensitive RH2 subfamilies, with members belonging to each subfamily linked in tandem within the genome (two SWS2-, and three RH2A genes, respectively). The presence of multiple cone opsin genes indicates that there have been duplication events in the cod ancestor SWS2 and RH2 opsins producing paralogs that have been retained in Atlantic. Our results are supported by expressional analysis of cone opsins, which further revealed an ontogenetic change in the array of cone opsins expressed. These findings suggest life stage specific programs for opsin regulation which could be linked to habitat changes and available light as the larvae is transformed into an early juvenile. Altogether we provide the first molecular evidence for color vision driven by only two families of cone opsins due to gene loss in a teleost.  相似文献   

12.
Chinen A  Hamaoka T  Yamada Y  Kawamura S 《Genetics》2003,163(2):663-675
Zebrafish is becoming a powerful animal model for the study of vision but the genomic organization and variation of its visual opsins have not been fully characterized. We show here that zebrafish has two red (LWS-1 and LWS-2), four green (RH2-1, RH2-2, RH2-3, and RH2-4), and single blue (SWS2) and ultraviolet (SWS1) opsin genes in the genome, among which LWS-2, RH2-2, and RH2-3 are novel. SWS2, LWS-1, and LWS-2 are located in tandem and RH2-1, RH2-2, RH2-3, and RH2-4 form another tandem gene cluster. The peak absorption spectra (lambdamax) of the reconstituted photopigments from the opsin cDNAs differed markedly among them: 558 nm (LWS-1), 548 nm (LWS-2), 467 nm (RH2-1), 476 nm (RH2-2), 488 nm (RH2-3), 505 nm (RH2-4), 355 nm (SWS1), 416 nm (SWS2), and 501 nm (RH1, rod opsin). The quantitative RT-PCR revealed a considerable difference among the opsin genes in the expression level in the retina. The expression of the two red opsin genes and of three green opsin genes, RH2-1, RH2-3, and RH2-4, is significantly lower than that of RH2-2, SWS1, and SWS2. These findings must contribute to our comprehensive understanding of visual capabilities of zebrafish and the evolution of the fish visual system and should become a basis of further studies on expression and developmental regulation of the opsin genes.  相似文献   

13.
All endodermal and mesenchymal cells of the sea urchin embryo descend from the vegetal plate, a thickened epithelium of approximately 50 cells arising at the early blastula stage. Cell types that derive from the vegetal plate are specified conditionally by inductive interactions with underlying micromeres, but the molecular details of vegetal-plate specification remain unresolved. In a search for regulatory proteins that have roles in vegetal-plate specification, a screen was performed to clone Krüppel/Krox-related genes from a Strongylocentrotus purpuratus embryo cDNA library. One newly identified clone, named SpKrox1, contained four zinc fingers and a leucine zipper domain. SpKrox1 expression was low in unfertilized eggs, increased severalfold to the early blastula stage and decreased between the early gastrula and pluteus stages. SpKrox1 mRNA was first seen in macromeres of 16-cell stage embryos and was restricted to cells of the developing vegetal plate thereafter. Vegetal-plate expression corresponded to a ring of cells around the blastopore and overlapped the expression patterns of other genes with potential roles in vegetal plate-specification. As the vegetal-plate cells invaginated into the blastopore, SpKrox1 expression was lost, suggesting that its role was not in endoderm differentiation per se but rather in the initial establishment of the vegetal plate.  相似文献   

14.
15.
Matsumoto Y  Fukamachi S  Mitani H  Kawamura S 《Gene》2006,371(2):268-278
A variety of visual pigment repertoires present in fish species is believed due to the great variation under the water of light environment. A complete set of visual opsin genes has been isolated and characterized for absorption spectra and expression in the retina only in zebrafish. Medaka (Oryzias latipes) is a fish species phylogenetically distant from zebrafish and has served as an important vertebrate model system in molecular and developmental genetics. We previously isolated a medaka rod opsin gene (RH1). In the present study we isolated all the cone opsin genes of medaka by genome screening of a lambda-phage and bacterial artificial chromosome (BAC) libraries. The medaka genome contains two red, LWS-A and LWS-B, three green, RH2-A, RH2-B and RH2-C, and two blue, SWS2-A and SWS2-B, subtype opsin genes as well as a single-copy of the ultraviolet, SWS1, opsin gene. Previously only one gene was believed present for each opsin type as reported in a cDNA-based study. These subtype opsin genes are closely linked and must be the products of local gene duplications but not of a genome-wide duplication. Peak absorption spectra (lambda(max)) of the reconstituted photopigments with 11-cis retinal varied greatly among the three green opsins, 452 nm for RH2-A, 516 nm for RH2-B and 492 nm for RH2-C, and between the two blue opsins, 439 nm for SWS2-A and 405 nm for SWS2-B. Zebrafish also has multiple opsin subtypes, but phylogenetic analysis revealed that medaka and zebrafish gained the subtype opsins independently. The lambda and BAC DNA clones isolated in this study could be useful for investigating the regulatory mechanisms and evolutionary diversity of fish opsin genes.  相似文献   

16.
Cichlid fish of the East African Rift Lakes are renowned for their diversity and offer a unique opportunity to study adaptive changes in the visual system in rapidly evolving species flocks. Since color plays a significant role in mate choice, differences in visual sensitivities could greatly influence and even drive speciation of cichlids. Lake Malawi cichlids inhabiting rock and sand habitats have significantly different cone spectral sensitivities. By combining microspectrophotometry (MSP) of isolated cones, sequencing of opsin genes, and spectral analysis of recombinant pigments, we have established the cone complements of four species of Malawi cichlids. MSP demonstrated that each of these species predominately expresses three cone pigments, although these differ between species to give three spectrally different cone complements. In addition, rare populations of spectrally distinct cones were found. In total, seven spectral classes were identified. This was confirmed by opsin gene sequencing, expression, and in vitro reconstitution. The genes represent the four major classes of cone opsin genes that diverged early in vertebrate evolution. All four species possess a long-wave-sensitive (LWS), three spectrally distinct green-sensitive (RH2), a blue-sensitive (SWS2A), a violet-sensitive (SWS2B), and an ultraviolet-sensitive (SWS1) opsin. However, African cichlids determine their spectral sensitivity by differential expression of primarily only three of the seven available cone opsin genes. Phylogenetic analysis suggests that all percomorph fish have similar potential.  相似文献   

17.
为了研究许氏平鲉(Sebastes schlegelii)雌鱼体内受精后仔鱼开口前和仔鱼开口后两个阶段氨基酸和脂肪酸的组成变化规律, 采用生化常规方法定量测定并分析了许氏平鲉发育早期的受精卵(FE)、胚胎期(ES)、初产仔鱼(PL1)、前仔鱼期(PL2)、后仔鱼期(PL3)和稚鱼期(J)6个阶段的氨基酸和脂肪酸组成特点及含量变动。结果表明: 总氨基酸含量从FE至PL1显著下降, 至PL3显著上升, 至J又显著下降(P<0.05); 游离氨基酸含量以FE最低(12.77 mg/g), 从FE到PL1显著上升(P<0.05), 并在PL1含量达到最高值(92.19 mg/g), PL1发育到J呈现先下降后上升再下降的变化趋势(P<0.05), 游离氨基酸与总氨基酸的比值范围在2.37%—19.66%。在各发育阶段干样中检出碳链长度在C14-C24的29种脂肪酸, 分别为9种饱和脂肪酸(SFA)、9种单不饱和脂肪酸(MUFA)和11种多不饱和脂肪酸为(PUFA), 受精卵中主要脂肪酸依次为C22:6n-3(DHA)、C18:ln-9c、C16:0和C20:5n-3(EPA)。胚胎期(FE-ES)的脂肪酸利用率顺序为SFA、MUFA、n-6PUFA、n-3PUFA, 主要以C18:3n-3、C18:0、C16:1n-7及C20:5n-3(EPA)作为胚胎期的能量来源, C22:6n-3(DHA)的实际利用率最低(9.71%), 被优先保存下来, C16:0的实际利用量最高(10.94mg/g); 仔鱼内源营养阶段(ES-PL1)脂肪酸利用率顺序为MUFA、n-6PUFA、SFA、n-3PUFA, 主要以C16:1n-7、C18:0、C20:4n-6(ARA)及C18:1n-9c作为开口前仔鱼的主要能量来源, 其中仔鱼对DHA实际利用量最高(18.23 mg/g)。PL1-PL3阶段DHA相对于EPA和ARA被选择性利用; PL3至J阶段ARA相对于EPA和DHA被选择性利用。研究表明: 许氏平鲉仔鱼开口前阶段总氨基酸含量与游离氨基酸含量的变化趋势截然相反, 胚胎期与仔鱼内源营养阶段脂肪酸利用率和利用量均有所不同, 仔鱼期DHA优先被利用, 过渡至稚鱼期ARA优先被利用。建议在仔鱼开口后添加富含DHA生物性饵料, 仔鱼过渡到稚鱼期在配合饵料中添加ARA营养物质, 防止苗种营养不足, 保证成活率。  相似文献   

18.
19.
为了探讨高原硬骨鱼类胚胎型血红蛋白的转换表达模式, 研究利用转录组数据和克隆方法研究了黄河裸裂尻鱼(Schizopygopsis pylzovi)胚胎型血红蛋白的组成和基因结构, 并通过qRT-PCR和整胚原位杂交方法确定了胚胎发育过程中血红蛋白基因的表达情况。结果表明, 黄河裸裂尻鱼胚胎型血红蛋白基因由hbae1、hbae4、hbbe1和hbbe3组成, 均包含3个外显子和2个内含子, 其中hbbe1/hbae1以“头对头”(3′-5′—5′-3′)转录方式串联在一起, hbbe3/hbae4则以“尾对尾”(5′-3′—3-5′)的转录方式串联在一起。进一步的研究初步判断hbae3、hbae5和hbbe2基因在裂腹鱼亚科鱼类进化过程中可能发生了丢失。qRT-PCR和整胚原位杂交结果均表明在早期胚胎发育过程中4个胚胎型血红蛋白基因的表达都是从受精后第5天(120 hpf)开始明显上升, 但是hbae4和hbbe3基因的高表达只维持在受精后第5天(120 hpf)至破膜期(216 hpf), 随后急剧下降, 而hbae1和hbbe1基因表达从破膜(216 hpf)开始急剧上升。研究揭示了硬骨鱼类一种全新的血红蛋白转换表达模式, 这可能与黄河裸裂尻鱼特殊的进化历程和高原环境的适应有关, 研究结果将为进一步深入研究硬鱼类血红蛋白的时序转换表达和环境适应机制奠定基础。  相似文献   

20.
Atlantic halibut eggs and yolk-sac larvae were incubated at 1, 5 and 8° C. Eggs incubated at 8° C gave slightly shorter larvae at hatching with a significantly smaller total cross-sectional area of white muscle fibres than eggs incubated at 5° C. Transport of eggs 2 days prior to hatching gave significantly longer larvae at hatching with a significantly larger red fibre cross-sectional area than when eggs were transported shortly after the blastopore closure. A higher survival until 230 degree days after hatching was also observed in the former group. All eggs incubated at 1° C died before hatching and all larvae incubated at 1° C died before 45 degree days after hatching. From hatching until 230 degree days the total white cross-sectional area increased threefold in all temperature groups. The increase in white cross-sectional area was entirely due to hypertrophy between hatching and 150 degree days (10 mm L S). Recruitment of new white fibres increased in germinal zones at the dorsal, ventral and lateral borders of the myotome from 150 degree days onwards, but at 230 degree days (12–13 mm L S) the recruitment fibre zone constituted <10% of the total white cross-sectional area. Larval incubation at 8° C gave slightly longer larvae with a significantly larger cross-sectional area of recruitment fibres at 230 degree days than incubation at 5° C. The larval group incubated at 8° C also had a significantly lower survival until 230 degree days than did the 5° C group. Incubation temperature regimes did not affect the volume density of myofibrils in the axial muscle fibres at 230 degree days. Thus hypertrophy is the predominant mechanism of axial white muscle growth in Atlantic halibut yolk-sac larvae and an increased rearing temperature during the yolk-sac stage increases white muscle fibre hyperplasia.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号