Aluminum induced crystallization of strongly (111) oriented polycrystalline silicon thin film and nucleation analysis

A polycrystalline silicon thin film was fabricated on glass substrate by means of aluminum induced crystallization (AIC). Al and alpha-Si layers were deposited by magnetron sputtering respectively and annealed at 480A degrees C for 1 h to realize layer exchange. The polycrystalline silicon thin film was continuous and strongly (111) oriented. By analyzing the structure variation of the oxidation membrane and lattice mismatch between gamma-Al2O3 and Si, it was concluded that aluminum promoted the formation of (111) oriented silicon nucleus by controlling the orientation of gamma-Al2O3, which was formed at the early stage of annealing.

棘蛙族(Tribe Paini)精子形态结构及精巢年周期研究

棘蛙族(Tribe Paini)隶两栖纲(Amphibia)、无尾目(Anura)、蛙科(Ranidae)、叉舌蛙亚科(Dicroglossinae)，由棘蛙属(Paa)、倭蛙属(Nanorana) 和沙巴蛙属(Chaparana)构成（Dubois,1992）。由于特殊的形态特征和染色体核型，棘蛙族受到国内外学者的广泛重视和研究，但是到目前为止，棘蛙族的系统发育关系尚未明晰，族下属种的分类和归属问题还有待进一步研究和新的证据出现。本文通过光学显微镜、电子显微镜和石蜡切片对棘蛙族10 物种的精子和精巢进行研究，旨在了解棘蛙族精子的形态、量度、超微结构特征及不同季节精巢结构的变化规律，同时为棘蛙族的系统研究提供新的依据，也为棘蛙族濒危物种的保护和经济物种的繁殖提供基础资料。研究结果表明：棘蛙族各属物种精子的形态基本相似，精子整体呈线形，由头部、中片和尾部构成。精子头部呈长条状，顶体呈锥状，位于头部顶端并向前伸出，中片较长，尾部波动弯曲。棘蛙族各属物种精子量度差异较大，将各属物种精子头部、中片、尾部、头宽、尾宽的量度数据进行聚类分析，结果表明棘蛙族10 物种可分为三类：第一类包括棘侧蛙、合江棘蛙、小棘蛙、棘腹蛙和棘胸蛙，特点是精子较短，全长在72.6～103.35µm 之间；第二类包括倭蛙、高山倭蛙、腹斑倭蛙，特点是精子较长，全长在107.74～129.75µm 之间；第三类包括隆肛蛙和双团棘胸蛙，特点是精子最长，全长在145.89～165.84µm 之间。棘蛙族各属精子超微结构基本相似：精子头部由顶体、细胞核构成；中片由中心粒、线粒体构成；尾部由单根轴丝构成。精子顶体横切呈圆环状，细胞核电子密度高；线粒体为卵圆形，呈环状围绕轴丝排列，线粒体数目较多，约30层；尾部轴丝为典型的9＋2结构，即由2根中央微管和9对外周微管组成。不同季节的倭蛙精巢结构变化表明倭蛙精巢每年只有一个生精周期，生精周期始于7 月，繁殖季节从5 月到6 月，生精高峰期为9 月；根据倭蛙不同季节精巢结构的变化，可将生精周期分为3 个阶段：第一阶段从7 月到9 月，为精子形成期；第二阶段从10 月到翌年4 月，为精子的贮存阶段，也即倭蛙的冬眠期；第三阶段从5 月到6 月，为精子的排放阶段，即倭蛙的繁殖期。不同季节的隆肛蛙精巢结构变化表明5 月为隆肛蛙的繁殖高峰期。根据棘蛙族各属精子的形态、量度和超微结构特征，结合已有的棘蛙族形态学、生态学、染色体核型及系统学研究成果，本文认为：1．基于精子数据对棘蛙族的划分和基于形态学及分子系统学数据对棘蛙族的划分均有相同之处，精子形态结构可为棘蛙族的系统研究提供新的证据。2． 棘蛙族各属精子的形态、量度及超微结构不仅与蛙科其他属种有明显差异，而且在无尾类中也较为特殊，精子学研究结果支持将棘蛙族从蛙科中分离出来，归隶于叉舌蛙科的叉舌蛙亚科的系统学修正。3. 精子的顶体、细胞核、中片的形态结构及量度可作为蛙科的分类指标。On the base of unique morphological and kyrotype characters, Dubois(1992)recognized three genera Paa, Narnorana, Chaparana as tribe Paini, which is amember of Dicroglossinae, Ranidae. In present study, the sperm shape, size andultrastructure of 10 paini species were investigated through the light and electronmicroscope, and testis structure of N. pleskei and F. quadrana was also studied. Wesuppose this study could offer some spermatological evidence to phylogeny andreproduction study of tribe Paini. The results were as follows:The sperm shape of tribe paini is homologically similar, the spermatozoa arefiliform, composed of elongate head, long mid-piece and waved tail. The acrosome isapically associated with the nucleus and extend anteriorly.The sperm length of tribe paini differ remarkably among genera. Cluster for thelength of sperm head, mid-piece, tail, total length, head-width, tail-width of ten painifrogs indicated the 10 species could be separated into three groups: GroupⅠcontainsP. shini, P. robertingeri, P. spinosa, P. exilispinosa, P. boulengeri, the spermatozoa ischaracterized with short in total length, ranging from 72.6µm to 103.35µm; GroupⅡcontains N. pleskei, N. parkeri, N. ventripunctata, the spermatozoa ischaracterized with relatively long in total length, ranging from 107.74µm to129.75µm; Group Ⅲ contains F. quadrana and P. yunnanensis, the spermatozoa is characterized with longest in total length, ranging from 145.89µm to 165.84µm. thethree groups based on spermatological data is partially match the classification basedon morphological and molecular data.The ultrastructure of spermatozoa in tribe paini is also basic similar, includingacrosome vescile, nuleus of the head proper, centriole, mitochondriol of themid-pieces, axoneme of the tail. The acrosome vescle is circle in TEM transversesection, the density of nucleus is high; The mitochondrions is oval, surrounding theaxial filament with about 30 layers of mitochondria; The axoneme has the typical 9+2pattern of microtubules.The seasonal changes in testis of N. pleskei indicates it has only onespermatogenesis circle, which begin in July, the reproduction season is from May toJune, the spermatogenesis is active in September. On the base of seasonal changes intestis, the spermatogenesis circle can be separated into three stages: In stageⅠfromJuly to September, spermatids are formed; In stage Ⅱ from October to April next year,the spermatozoa are stored in testis,which is the hibernated period; In stage Ⅲ fromMay to June, mature spermatozoa were released from the testis, which is thereproduction season of N. pleskei. As to F. quadrana, reproduction is active in May.With the previous study of morphology, ecology, karyotypes and phylogenyresearch of tribe Paini, the spermatological data in present study suggests:1. The spermatological classification of tribe paini is partially consistant with themorphological and molecular classification respectively.2.The sperm morphology and ultrustructure of tribe paini is unique not only inthe family Ranida but also in Anura, which suggest the tribe paini is monophyletic andmight be transfered from the family Ranida to the family Dicroglossidae based onmolecular evidence.3. The acrosome, nuleus, shape, length and ultrastructure of mid-piece can beused as an alternative taxonomic character in Anura.

重离子诱导人舌鳞癌细胞凋亡和Bax/Bcl-2蛋白表达的变化

In order to investigate the effect of carbon ion irradiation on apoptosis and Bax/Bcl-2 expression inhuman tongue carcinoma cells, exponentially growing human tongue carcinoma cells (Tb) cultured in vitro were irradiated with 0, 0.5, 1.0, 2.0 or 4.0 Gy of 12C6+ ions respectively. Survival rate of irradiated cells at various doses were measured by MTT assay. The nucleus changes of apoptosis and necrosis of cells stained by Hochest/PI were observed through fluorescence microscope. The cell cycle changes were detected by flow cytometry (FCM). The expressions of Bax and Bcl-2 were detected by Western blot analysis. The results show that the viability of Tb cells decreases gradually with increment of irradiation doses of carbon ions. The proportions of apoptosis cells in the irradiated groups are significantly higher than those in the control group. There is a positive correlation between irradiation doses and retardation strength in G2 /M phase at 24 h after irradiation (P<0.05). And the expressions of Bax and bcl-2 are significantly up-regulated and down-regulated respectively by 12C6+ ion irradiation. It can be concluded from above that cell apoptosis induced by heavy ion with high-LET may be mediated through the Bax/Bcl-2 expression pathway. 探讨重离子辐照对人舌鳞癌Tb细胞的凋亡及Bax/Bcl-2蛋白表达的影响。采用0、0.5、1.0、2.0、4.0 Gy重离子束辐照人舌鳞癌 Tb 细胞，应用 MTT 法检测细胞存活，流式细胞技术检测细胞周期变化，Hoechst33258/PI 复染法观察 Tb 细胞凋亡形态，并采用 Western-blot 法检测 Bax/Bcl-2 蛋白表达情况。结果发现，Tb细胞经12C6+离子束辐照后存活率显著下降，呈剂量依赖性的生长抑制；Tb细胞呈现蓝色荧光浓集成团的凋亡形态，且凋亡比例随辐照剂量增加；G2/M 期细胞百分数随照射剂量增加而增加（P＜0.05） 。Western-blot结果显示 Bax 蛋白表达水平随辐照剂量逐渐上升，但在 4 Gy 组其表达不再增高，Bcl-2 蛋白在 1.0、2.0、4.0 Gy组随剂量增大呈下降趋势。以上结果提示重离子束辐照对 Tb 细胞有抑制作用，Bax/Bcl-2 蛋白表达是重离子治癌的机制之一。