为明确威宁中华蟾蜍的分子分类学地位，对采自贵州省威宁县的蟾蜍标本（标本号：LPSWN01A）进行线粒体12S rRNA 基因扩增和双重单系法分析。结果表明：该标本的序列大小为906 bp （GenBank 登录号：KJ803016），基于数据集 A 构建的系统发育树中，威宁中华蟾蜍与7条中华蟾蜍同源序列聚为一个单系（中华蟾蜍支系），威宁中华蟾蜍以较低的最大似然率独立为一个支系；基于数据集 B 构建的系统发育树中，威宁中华蟾蜍与2个中华蟾蜍华西亚种种群聚为一支，因此推断威宁产蟾蜍属于中华蟾蜍华西亚种（Bufo gararizans andrewsi ）。另外，威宁中华蟾蜍与云南怒江种群的遗传距离最近，为0．0093，说明威宁中华蟾蜍属于中华蟾蜍华西亚种，与之前的传统分类文献记载一致。

The mitochondria 12S rRNA gene of Weining B.gargarizans sample (LPSWN01A)was amplified and analyzed by dual monophyletic method to definite molecular taxonomic status of B. gargarizans.The sequence length of Weining B.gargarizans is 906bp (GenBank accession number:KJ803016).The sequence of Weining B.gargarizans and seven homologous sequences of B.gargarizans form a monosystem in the phyogenetic tree built by data set A. Weining B.gargarizans forms an independent clade with lower maximum likelihood. Weining B.gargarizans and two B.gargarizans andrewsi populations form a monosystem in the phyogenetic tree built by data set B,which implies that Weining B.gargarizans belongs to B.gargarizans andrewsi .In addition,the genetic distance between Weining B.gargarizans and Nujiang River population is 0.009 3 only,which proves that Weining B. gargarizans belongs to B.gargarizans andrewsi also.In conclusion,the molecular identification result of Weining B.gargarizans is in accordance wit

目的 探讨蟾蜍灵联合顺铂对舌鳞状细胞癌（以下简称舌鳞癌）Tca8113细胞增殖、凋亡的影响及其机制。方法 培养人舌鳞癌Tca8113细胞株,取对数期细胞分为对照组、蟾蜍灵组、顺铂组、联合组。对照组加入RPMI 1640培养基;蟾蜍灵组分别加入10、20、40、80、160 nmol/L的蟾蜍灵;顺铂组分别加入2.5、5、10、20、40μg/m L的顺铂;联合组分别加入10、20、40、80、160 nmol/L的蟾蜍灵后按浓度对应加入2.5、5、10、20、40μg/m L的顺铂;MTT法检测各组细胞增殖抑制率,计算抑制细胞增殖50%的药物浓度（IC50）及联合指数（CI）。根据蟾蜍灵与顺铂48 h时的IC50选择三个药物组相应浓度亚组,即蟾蜍灵40 nmol/L组（蟾蜍灵亚组）,顺铂10μg/m L组（顺铂亚组）,蟾蜍灵40 nmol/L、顺铂10μg/m L组（联合亚组）,流式细胞术分析各亚组细胞凋亡情况,Western-Blot法检测各亚组Bax、Bcl-2、Caspase-3、细胞外调节蛋白激酶（Erk）及其磷酸化细胞外调节蛋白激酶状态（P-Erk）的表达。结果 与对照组相比,蟾蜍灵组、顺铂组、联合组细胞增殖受到抑制,且联合组增殖抑制率高于蟾蜍灵组与顺铂组,并呈时间-剂量依赖关系（P均〈0.05）。联合组联合指数（CI）均小于1。联合亚组细胞凋亡率高于蟾蜍灵亚组与顺

Objective To investigate the effects of bufalin and cisplatin on proliferation and apoptosis of tongue squa -mous cell line Tca8113.Methods Tongue squamous cell line Tca8113 was cultured and the logarithmic phase cells were divided into control, bufalin(10,20,40,80,160 nmol/L), cisplatin(2.5,5,10,20,40 μg/mL) and combination groups. MTT assay was used to examine the inhibition rate of cell growth when cell were treated at various concentrations of bufalin and cisplatin alone or in combination and calculate inhibiting concentration of 50%cell growth ( IC50 ) .The corresponding concentrations of three drug groups were selected based on IC 50 of Bufalin and cisplatin at 48 h.Apoptosis was evaluated by flow cytometry.The protion expression of Bax ,Bcl-2,Caspase-3,P-ERK and Erk were examined by Western blot method . Results Bufalin or cisplatin effectively inhibited the growth of Tca 8113 cells in a concentration-dependent manner at cer-tain range of concentrations .The inhibitor

汉代画像石中的西王母仙境中有许多蟾蜍图像,它们成为汉代人企盼永生不死、羽化升仙的重要印证。在西王母仙境中,蟾蜍的构图方式以动态形式出现,与西王母的文静雍容成对比,使仙境图像更为丰富而具有张力。从神话思维、民俗学以及汉代的阴阳五行诸多方面揭示蟾蜍物象作为汉代艺术升仙符号的重要文化内涵。

There are a great number of toad images in the portrait of West Queen from Han carved stones.They are regarded as important proofs to testify people’s longing for longevity and immortality in the Han Dynasty.In the fairyland of West Queen,the toads are always in motion,forming a contrast with the graceful and reposeful Queen and making the fairyland more vivid and colorful.Approaching these toad images from mythological logic, folklore,yin,yang and the five elements of earth,this paper intends to unveil their cultural connotations and their symbolic meaning of longevity and immortality in the Han Dynasty.

关于抗癌活性的蟾蜍二烯内酯的结构修饰化合物已达到100多个。甾体母核3位磺酸化的蟾蜍二烯内酯是从蟾蜍皮中分离得到的一类具有抗癌活性的微量成分,但目前尚无关于蟾蜍二烯内酯磺酸化修饰的报道。该文以沙蟾毒精为例,首次发展了一种高效的3位磺酸化沙蟾毒精的合成方法,对目标产物进行了结构鉴定。基于自制亲水性材料Click TE-Cys,发展了反相/亲水串联固相萃取的反应产物后处理方法,用于磺酸化沙蟾毒精的富集。依此法对总蟾毒配基进行了磺酸化和目标产物的富集及LC-MS定性分析,实现了大量具有潜在抗癌活性的新化合物的制备和富集,为进一步实现高效低毒化合物的筛选提供了物质基础。

Bufadienolide is a kind of natural steroids with significant anti-cancer activities. Hitherto, the number of structural modification on the bufadienolides has reached more than 100 . The purifica-tion of sulfated,a trace cytotoxic component bufadienolides from toad skin is a tough work due to its low content and unstable structures. However,there is no report on synthesis of bufadienolides sul-fonation though they have promising cytotoxic activities. In this paper,the arenobufagin was firstly taken as an example,to develop an effective method for the synthesis of arenobufagin-3-O-sulfonate. In order to enrich the arenobufagin-3-O-sulfonate quickly and mildly,a reversed phase/hydrophilic solid-phase extraction method based on our homemade hydrophilic Click TE -Cys was developed. The structures of arenobufagin-3-O-sulfonate and arenobufagin were both identified by MS,1 H NMR and 13 C NMR spectra. In addition,the total bufogenins were sulfonated by this method and the sul-fated bufogenins

以骨髓细胞作为材料制备染色体标本，对福建宁德黑眶蟾蜍(Bufo melanostictus)的染色体核型及银染进行观察与分析。结果表明：染色体数为2n=22(20M+2SM)，NF=44，配成11对染色体，由6对大型染色体和5对小型染色体组成，未发现与性别分化相关联的异型染色体，染色体未见银染区域。与已报道的广东广州、福建福州、广西南宁、浙江温州、云南腾冲、台湾等不同地域的黑眶蟾蜍核型等研究资料进行比较，发现不同产地的黑眶蟾蜍核型在SM数目和顺序、次缢痕或随体的位置、银染区域等方面都存在着一定的差异。

The observations and analysis of Bufo melanostictus karyotype, Ag-NORs were based on the chromosome sample from the bone marrow cells. The results showed that:the number of chromosome was 2n=22 (20M+2SM), NF=44, with 11 pairs of chromosomes consisting of 6 pairs of megachromosome and 5 pairs of minisome. No heteromorphic chromosome was found correlating with sex differentiation. No region of the Ag-NORs chromosome was found. Compared with the reported research about B. melanostictus karyotype from Guangzhou Guangdong, Fuzhou Fujian, Nanning Guangxi, Wenzhou Zhejiang, Tengchong Yunnan, Taiwan and other different regions, it showed that there were some differences about B. melanostictus karyotype from different regions in the SM number and the order, secondary constriction and satellite, Ag-NORs areas and so on.