線蟲補捉菌Arthrobotrys musiformis 黏液相關基因之選殖與功能界定
線蟲捕捉菌Arthrobotrys musiformis 是一種可經線蟲誘導產生捕捉網來捕捉線蟲的真菌，本實驗即針對A. musiformis 的捕捉網黏液相關基因：Manosyltransferase（AH73）, β-1,3-glucan transferase（AH102）, fimbrin（AH121）及mannose-specific lectin precursor（AH338）進行選殖與功能界定，希望建立這方面的研究基礎，將來能應用在松材線蟲的生物防治上。首先我們大量培養A. musiformis，萃取菌絲體的DNA；接著進行聚合?連鎖反應 (Polymerase Chain Reaction，PCR) ，利用專一性引子對 (primer) 大量增幅AH73、AH102、AH121 及AH338之基因片段；增幅後的產物經過純化、選殖，定序並進行分析比對，確認增幅之序列無誤後，以 Digoxigenin (DIG) 標示當為探針，篩檢A. musiformis 的Fosmid Library﹔目前已成功選殖出AH73 之可能基因，完成AH73 之探針製備，並以其篩檢A. musiformis 的Fosmid Library﹔呈雜合正反應之選殖株 (clones) 將以散彈槍方法（shotgun）定序，作序列組合，探索相關的基因；接下來用 Rapid Amplification of cDNA Ends（RACE) 做出互補DNA (complementary DNA , cDNA) 全長度後；最後建構基因缺失株，驗證此基因所調控的生理以及生化機能。 Nematode trapping fungus Arthrobotrys musiformis can capture nematodes by producing adhesive nets when nematodes go through. Many kinds of nematodes, including pine wood nematode (Bursaphelencus xylophilus), can be captured. Pine wood nematode causes serious pine wood disease. Therefore, A. musiformis has the potential of biocontrol in pine wood nematode. Our research focused on adhesion and adhesive relevant genes of A. musiformis ：Manosyltransferase (AH73), β-1,3-glucan transferase (AH102), fimbrin (AH121), and mannose-specific lectin precursor (AH338). We try to clone these genes and carry out functional analysis. In order to achieve this goal, we used specific primers derived from previously obtained complementary DNA (cDNA), by Polymerase Chain Reaction (PCR) to amplify these genes and gained adequate quantity of genomic DNA products. After sequencing and verifying of the identity of the genomic DNA, we use Digoxigenin （DIG） to label them and use them as probes to screen the constructed A. musiformis Fosmid Library. Currently, the Southern colony hybridization is undergoing. The positive Fosmid clones against the specific probes will be sequenced completely by shotgun library to monitor the existence of adhesion related gene cluster. After working out the full length cDNA of these genes, we will use them to construct replacement vectors to knockout the adhesion related genes, creating mutants and further verify their functions through genotype or phenotype bioassay.
生生不息-正五邊形的繁衍法則
This study was to explore the nature of two basic constitutes of the regular pentagon,With these two constitutes, the regular pentagon could be multiplied into any times. We used four multiplication methods (m2 = 2m1 + n1 、n2 = m1 + n1 、m2= k2m1 、n2= k2n1、a2 = a1 + 1、a2 = a1 + ) to show how the regular pentagon could enlarge and to verify that the enlarged regular pentagons derived from computer did exist. By integrating these four multiplication methods, we were able to arrange regular pentagon of any length of side, and evidenced the equation was
( If the side length of a regular pentagon is a form of m,n is the number of A,B respectively )
We further proved that the first multiplication method could be developed into a new modified method, which could divide a regular pentagon with a given side length into a combination of A and B. But only when the x and y of side length of a regular pentagon could be divided by a natural number, k, and made x/k into an item of the Fibonacci Sequence and y/k a successive item.
When we tried to verify if any regular pentagon could be constituted by other smaller regular pentagons, we also found that it was un-dividable only if the length of pentagon side were ( the number of A, B were the 2n and 2n-1 item of Lucas Sequence). Otherwise, any regular pentagon might be able to be constituted by other smaller regular pentagons.
本研究是以正五邊形的兩個基本組成元素(B)作為討論對象，利用此二元素可以將正五邊形做任意倍數的放大。我們共使用4種繁殖法則(m2 = 2m1 + n1 、n2 = m1 + n1 、m2= k2m1 、n2= k2n1、a2 = a1 + 1、a2 = a1 + ) 來說明正五邊形的放大情形，並利用此4 種繁殖法驗證電腦運算出的放大圖形確實存在。利用這4 種繁殖法則的改良與整合，已達到能排出任意邊長之正五邊形的目標，並能計算並證明出其通式為。
(若正五邊形的邊長為形式,m、n代表、的個數)
更特別的是，我們能用第一繁殖法反推出一種方法，將給定邊長的正五邊形利用簡單的切割方式分成由A、B 組合成的形式，但只有正五邊形邊長之x、y 值可同除以任一自然數k 而使 x/k 為費波那契數列之一項且 y/k 為其後一項者才可以使用。
將此想法推廣至一個正五邊形能否由比他小的其他五邊形組合而成時，我們也發現當正五邊形之邊長為時（其A、B 個數為盧卡斯數列之第2n,2n-1 項），不可分解，否則應該皆可將一個正五邊形分解成比它小的其他五邊形組合（我們也可以利用這些質形檢驗出其他正五邊形是否也為質形）。但其分解形式，不只一種，而我們推測只用兩種較小的正五邊形就能達成，我們期待能找出一或多種分解方法，能將正五邊形分解成標準的分解形式。
Mechanism of the subcellular localization of the actin binding protein adducin
Adducin蛋白在細胞骨架的調節上扮演著重要的角色。然而，近來有許多研究指出，骨架蛋白也會出現在細胞核並參與轉錄調控，因此本研究的目的即在探討adducin蛋白是否會進入細胞核中，並參與轉錄調控或具有其他功能。在本研究中，我們將綠色螢光蛋白(GFP)標示的adducin質體DNA，利用轉染技術送入老鼠纖維母細胞株NIH3T3中表現。NIH3T3細胞原本並無adducin蛋白的表現，在共軛焦顯微鏡下觀察，野生型的GFP-adducin蛋白會表現於細胞核與細胞質中。由於adducin蛋白尾端序列攜有可能往核內運輸的訊號，於是將位在此一訊號中的離胺酸718及離胺酸719進行突變，結果發現此一突變株只能在細胞質中表現。此外，蛋白磷酸脢C(protein kinase C)已知能磷酸化adducin蛋白在絲胺酸716及絲胺酸726的位置，於是假設其磷酸化是否與其在細胞內的分布有關。將adducin的絲胺酸726置換成丙胺酸，並不影響其在細胞內的分布。然而將絲胺酸716置換成丙胺酸後，則完全只在細胞核中表現。由於adducin可分布於細胞核，因此我們懷疑adducin蛋白可能與細胞分裂有關，於是本研究利用流式細胞儀分析adducin轉染後NIH3T3細胞的細胞週期。流式細胞儀的分析結果顯示，攜有GFP-adducin或其突變株的細胞與未經轉染的NIH3T3細胞的細胞週期並沒有顯著差異。其次，為了避免因轉染的效率不高而造成統計上的誤差，我們利用顯微鏡追蹤技術觀察攜有GFP-adducin的細胞株，結果顯示攜有adducin突變株的NIH3T3細胞株仍能正常分裂。再者，因為adducin能與細胞骨架中的肌動蛋白結合，所以adducin不同的分布位置可能影響細胞附著與細胞展延的效率。細胞展延試驗的結果顯示，adducin及其突變株對細胞附著與細胞展延的效率並無明顯的影響。本研究的結果證明，adducin的確帶有往核內運輸的訊號，其在細胞質中的分布可能也同時受到絲胺酸716磷酸化的影響。然而adducin的功用似乎與纖維母細胞的分裂與展延無明顯的關聯性。Adducin, an actin binding protein, is known to play an important role in the regulation of the membrane cortical cytoskeleton. More and more evidence indicates that proteins involved in the cytoskeletal regulation could also reside in the nucleus and participate in gene regulation. Thus, the goal of this study is to examine whether adducin is expressed in the nucleus and involved in certain nuclear events. In this study, adducin and its various mutants were fused with green fluorescent protein (GFP) and transfected into mouse NIH3T3 fibroblasts which do not have endogenous adducin for monitoring their subcellular distribution under a laser scanning confocal microscope. The wild-type GFP-adducin was found to be present both in the nucleus and in the cytoplasm. The COOH-tail of adducin contains a motif analogous to the nuclear localization signal (NLS). Mutation of two lysine residues (lysine 718 and lysine 719) located within this motif abolished the nuclear localization of adducin. Moreover, adducin is known to be phosphorylated by protein kinase C at serine 716 and 726. Substitution of adducin serine 726 with alanine had no effect on its subcellular localization. In contrast, substitution of adducin serine 716 with alanine led to only nuclear expression. Nuclear localization of adducin renders it possible that adducin may be involved in the regulation of cell division cycle. For cell cycle analysis, flow cytometry was applied. The results of flow cytometry indicated that expression of adducin and its mutants in NIH3T3 fibroblasts did not affect their cell cycle progression. To further examine the effect of adducin on cell division, NIH3T3 cells transiently transfected by adducin were monitored by time lapse video microscopy. The video clearly showed that the cells with GFP-adducin underwent cell division to generate two daughter cells. Since adducin is well known to bind to actin and thereby regulate microfilaments, we wondered that expression of adducin in NIH3T3 cells might affect their adhesion and spreading onto extracellular matrix proteins. The results of cell spreading assays showed that adducin appeared not to affect cell spreading. In conclusion, our results demonstrate that the subcellular distribution of adducin is likely regulated by two signals, one is the nuclear localization signal and the other is the phosphorylation status of the serine 716. However, enforced expression of exogenous adducin in fibroblasts such as NIH3T3 cells does not alter their cell cycle or cell spreading on fibronectin.
Pleistocene mammals of a river basin of Adycha
The paper represents the results of study of remains of mammalians which inhabited the Adycha-River basin (Verkhoyansky Region) during the Pleistocene Epoch. All studied materials belong to the Museum of Paleontology and Ethnography of the Adychinskaya Secondary School. For a long time, People have been finding ancient animal bones on the Adycha-River banks, and since 1955, the teacher, Bozhedonov Nikolai Gerasimovich, has been organizing exploring local lore expeditions for school children purposed for mammoth fauna investigation and material collection for the school museum. In this research collected material was classified, measurements of the Pleistocene Period animal bones were made. On the basis of it we came to conclusion that during the Pleistocene Epoch, very rare species of animals inhabited the Adycha-River basin. To our opinion, the Adycha-River basin is a unique site for research activity. Comparative measurements of the Pleistocene Periods animal bones were made. Of flu work we know measurements of the Early and Middle Pleistocene Period elephant’s bones which inhabited the Adycha-River basin. Elephants inhabited with mammoth. To our opinion, they didn’t yield to size to mammoth. In that time the Adycha-River basin, was been inhabited by camelopard, bison and Soergelia.\r Museum has 2 skull’s fragments and ovibos fronts part of horny stream. Geological age which is Early Pleistocene or late Pleistocene. These bones concern to Praeovibos. Distance between horns is more 100 mm. If we compare with ovibos skull of the Late Pleistocene we see that distance between horns is longer for the 10 time. We couldn’t found such comparisons at scientist’s works. It results that the more distance between horns is longer the more ancient, and skull’s sizes are much less.\r 7-8 km lower of Betenkes, in disclosing “?????? ???????” and sand bank “??????”, was been founded 3 crag’s part of skull of Soergelia, metacarpal bone and 2 skull’s fragments with horns part. On the Adycha-River basin was bun founded above all quantity of soergelias remains.\r In the Pleistocene Period inhabited a lot of horses. Which remains he have now. Our horse’s height in crest is from 142 to 156 sm. And what about of horses of the Early Pleistocene, it is more than 230 sm.\r Molar teeth of the Early Pleistocene Epoch horse are more for the 2.5 times than of contemporaneous analog.
利用奈米色料製作彩色蠶繭之研究
由於奈米科技進步，奈米材料應用在產業上具有多功能的性質。本研究使用不同波美度的色料餵食家蠶，以找出最佳的彩色蠶繭色澤，並研究其如何影響家蠶所結出的蠶繭及色料附著在蠶繭上的絲。同時對色料附著的蠶繭進行水洗、光照、微結構的觀察，以試圖找出色料與波美度之最佳組合參數。由本實驗結果得知，利用奈米色料溶液60 ml，在紅色：1.048、藍：1.058、黃：1.039 的參數下，混合飼料30g，可獲得最佳的彩色蠶繭結繭成功率、均勻度較佳、耐褪色與耐洗滌等優點，並且可獲得表面結構光滑且較細的絲徑，約為19.87μm。相對的，一般色料粒徑為微米級，色彩度優於奈米色料，但表面結構較奈米色料粗糙且線徑較粗，約為21.51μm，易於褪色及不耐洗滌。 Because of the great progress of nano-technology, it has the quality of multi-functions to make use of nano-materials on industrial property. The purpose of this study is to find the best colored silkworm cocoons by mixing different consistency of pigments to feed silkworms. At the same time, this study wished to explore how the different consistency of pigments influenced the silkworm cocoons that the silkworms produced. Besides, in order to find the better association between pigments and Baume degrees, this study exposed the cocoons under different lights, washed with different detergents and take observations of micro-structure of the cocoons. The results of this study are as follows: using the nano-pigments 60 ml in different density, that is, red:1.048, blue:1.058, and yellow:1.039, then mixed them with silkworms’ forage 30g , in this way, best successful ratio to get colored cocoons, desired high visual effects in color, well distribution, long duration and strong resistance to detergents agents are obtained. In addition, we can still get glossy appearance and fine cocoons; the wire diameter is about the size of 19.87μm. On contrast, feeding with the ordinary pigments, the degree of colored silk is better than feeding with the nano-pigments, but the appearance of cocoons are rough and the wire diameter is about the size of 21.51μm.