黏質色拉雷菌(Serratia marcescens)發光重組菌偵測環境中含酚環之毒性化合物之?
A pair of bacterial two-component system RssB-RssA was cooperated into Serratia marcescens for toxicity phenolic compound detection. First step of this study, E coli was used to accept the plasmid and certified by fluorescent. Then transfer the system from E coli into Serratia marcescens. Finally, 7 kinds of chemical, included phenol, benzene, toluene, xylenes, 4-chlorotoluene, 2-nitrotoluene, and kerosene, were used to check the sensitivity of this gene modified Serratia marcescens line. The results showed that this gene modified Serratia marcescens line had good performances and responses to those chemicals. 本實驗是以一受到二元訊號傳遞系統調控的發光基因重組質體,送入黏質色拉雷菌中,並以製備好的菌株進行毒性化合物之測試。在實驗的第一階段,我們將重組質體送入大腸桿菌內,並以其發光的有無來判斷是否達到送入的目的,其後再以電泳法確認各基因片段是否正確。第二階段再以相同的方法將選殖好的重組發光質體送入黏質色拉雷菌。第三階段,以發光重組菌針對酚、苯、甲苯、二甲苯、4-氯甲苯、2-硝基甲苯及煤油進行發光測試。結果方面,我們發現黏質色拉雷菌發光重組菌對於這一系列的酚環類化合物的確具有相當高的敏感度。
台灣兒科病人罹患神經母細胞瘤者可檢測到微小病毒B19的存在
罹患神經母細胞瘤的兒科病人,尤其是罹患stage IVs 神經母細胞瘤者,他們有些伴隨著非常嚴重的貧血,但卻檢測不出神經母細胞瘤已經侵犯骨髓;有時病情來勢洶洶,尤其是腫瘤細胞中已可偵測到N-myc 基因增幅者,診斷時腫瘤細胞可能已在腹腔四處擴散並已侵犯大部分的肝臟。但是,某些這種病患,特別是腫瘤細胞中N-myc 基因沒增幅者,即使在沒有治療的狀況下卻可能有自然恢復的現象,也就是腫瘤細胞會自動消退,但原因仍待進一步的證實與探討。可是,這些病人在其病情最嚴重的時候,骨髓內紅血球母細胞形態上的改變顯示可能與病毒感染有關。但是關於病毒來源的研究,現有的資訊仍然十分有限,其中最重要的是,病毒感染與引發其後天之免疫作用是否有關,更需要深層的研究。因此,為更進一步了解罹患神經母細胞瘤之兒科病人的病毒感染及病毒蛋白表現的作用,我們這次研究的目的在檢驗罹患神經母細胞瘤及貧血之兒科病人與微小病毒B19 (PVB19)、Epstein-Barr Virus (EBV)、腸病毒71 型(EV 71)和巨細胞病毒(CMV)的關係,以及病毒蛋白表現對這些病人的作用與臨床意義。In pediatric patients with neuroblastoma, in particular, those with stage IVs neuroblastoma, sometimes the disease was combined with severe anemia. However, no tumor involvement was detected in the bone marrow. Although some of these patients may have N-myc gene amplification, and the disease could have invaded many abdominal organs, especially liver, interestingly, the disease might regress spontaneously in some of these patients. The medical reason of the spontaneous regression, nonetheless, remains to be determined. It is worth noting that morphological changes of erythroid progenitor cells in the bone marrow have suggested virus infection in these pediatric patients. However, the available information of viral origin is limited. Furthermore, it is possible that the virus infection in these patients could be associated with the revocation of immune responses related to the spontaneous regression of the tumor. In this study we will investigate the relationship of parvovirus B19 (PVB19), Epstein-Barr virus (EBV), enterovirus 71 (EV71) and cytomegalovirus (CMV) with neuroblastoma by PCR in Taiwanese pediatric patients. Moreover, we will study the effect and the clinical significance of viral gene expression as well as N-myc gene amplification in these patients.
凸n 邊形等分面積線數量之分布探索
(一) 本研究首先導出ΔABC等分面積線移動所包絡出的曲線方程式,其圖形是由等分面積線段PQ(其中P、Q皆在ΔABC的周界上)的中點所構成,具有3 條曲線段(分別為3 條雙曲線之一部分)的封閉曲線,形成內文所謂的「包絡區」。利用包絡區的區隔,我們找出:1.當P 點在包絡區內,則有3 條等分面積線。2.當P 點在包絡區周界上,則有2 條等分面積線。3.當P 點曲線段的端點或在包絡區外,則有1 條等分面積線。(二) 以三角形的研究當基礎,擴展到凸n 邊形(不包含點對稱圖形),我們發現:等分面積線數量之分布,仍然與包絡區息息相關,且1.凸2m +1邊形最多有2m +1條等分面積線。2.凸2m邊形,必發生內文所謂的「換軌」。因此,最多只有2m ?1條等分面積線。3.包絡曲線所分割出的區域,於相同區域其等分面積線數量相同,且相鄰兩區域數量差兩條。(三) 若凸n邊形有k個「換軌點」,則此n邊形過定點等分面積線至多有n ? k 條。(四) 若凸n 邊形為點對稱圖形(如正偶數邊形、平行四邊形),則所有等分面積線皆過中心點。1) Our study got a curve equation of bisectors of a triangle. When a bisector is moving, we get three curves. They’re constructed by the midpoints of PQ. The three parts of the three curves make a closed curve which we called “the Envelope Area”. We found out:\r 1. When Point P is in the Envelope Area, we can get 3 bisectors. 2. When Point P is on the curves of the Envelope Area, we can get 2 bisectors. 3. When Point P is outside of the Envelope Area, we can get only 1 bisector. 2) Based on our study of triangles, we found that in Convex polygons(not including Point Symmetry Convex polygons), the distribution of bisectors is related to the Envelope Area. 1. We can get at most 2m +1 bisectors in a 2m +1 Convex polygon. 2. We can get at most 2m ?1 bisectors in a 2m Convex polygon, and the bisectors on the curves will “Change the Track”. 3. Envelope curve will divide a Convex polygon into several areas. The same area has the same numbers of bisectors, and the near areas have less or more 2 bisectors. 3) If a Convex polygon has k points to change the track, it will have at most n – k bisectors.\r 4) In a Point Symmetry Convex polygon (ex. Regular 2m convex polygons and parallelograms), all the bisectors will come through the center point.
藉由lotus effect 之原理來探討超疏水性表面的製備
超疏水表面(superhydrophobic surface)因其自我清潔效果在近年來引起廣泛的研究及探討,目前世界各大公司皆積極投入自潔性產品(超疏水表面)的研發,但是截至目前為止除了塗料及織物有產品外,其他尚在研發階段。降低表面能及增加表面粗糙度為製作超疏水性表面的兩種方法。本實驗我們針對使用不同方法去仿照自然界植物「蓮花效應」,將二氧化矽粒子適當的分佈在基材表面,以增加粗糙度,製造出超疏水性表面。我們發現鍛燒會使其二氧化矽粒子黏聚在一起,對接觸角的增加無幫助。而在溶膠凝膠的配置過程中直接加入OTS(Octadecyltrichlorosilane),並利用光散射儀比較二氧化矽奈米顆粒的平均粒徑及粒徑分佈隨時間變化的情形,一段時間後發現會有明顯大顆粒沉澱,塗佈在玻璃基材表面後容易脫落,不適合往後實驗或其他用途。而以氨水催化的溶膠凝膠以不同流速滴入TEOS (Tetraethylorthosilicate,四乙氧基矽)對接觸角的影響也不大。最後我們利用砂紙在基材表面上刷磨,可以使二氧化矽粒子分佈均勻,明顯增加接觸角。塗佈速率的改變搭配疏水性矽烷單分子膜的改質,我們已可以製造出159°的超疏水性表面。Superhydrophobic surfaces are generally made by lowering the surface energy and increasing the surface roughness. In this experiment, we use different methods of spreading silicon dioxide nanoparticles properly on the surface in order to increase the surface roughness and also make superhydrophobic surfaces. In the beginning, we find that the calcinations can cause its silicon dioxide nanoparticles to stick together instead of increasing the contact angles. Then, add OTS (Octadecyltrichlorosilane) directly to manufacture process of sol-gel, and observe the situations of the average length and the spread of silicon dioxide nanoparticles with the time goes by. After a period of time, we will discover that many obvious big particles deposit and spin-coating on the glass surfaces flop easily. Hence, this phenomenon is n’t proper for the following experiments or other uses. However, the contact angles have nothing to do with dropping the sol-gel catalyzed ammonia to TEOS(Tetraethylorthosilicate) by different flowing rates. Finally, it is crucial for us to use the sandpapers to brush on the surfaces because it may cause the silicon dioxide nanoparticles to spread well and obviously increase the contact angles. Combining the silicon dioxide surfaces with the change of spin-coating rate and the cover with hydrophobic SAM, we have made the superhydrophobic surfaces of 159°.
有毛!沒毛!哪個好!?探討石田螺及其螺殼上附生藻類與環境因子之關係
This research is about two ponds in the B park’s and the D park’s snail(Square Mystery Snail:Sinotaia quadrata) in Taipei city of Nei-hu District for research object, carry out the study of the following research proceed: 1.Discriminate the algae species that are growth on the snail shell and which is a kind of interaction with the snail; 2.The influence of the snail and algae with difference of temperature, salinity, pH value and dark ; 3. The factors affect algae growth on snail shell; 4.Use the variation of snail and algae to be a biological incator. The result manifestation: the algae that are growth on snail shell have two kinds, one is Cyanophyta and the other is Cladophora sp. The interaction between algae and snail belong to communalism, but under the condition of lacking of food, the snail then will eat the Cladophora sp. which grow on the shell of other snails. The temperature adapts aspect, upper limit of the feat existence of the snail should be low in 28℃. When over than 28℃, Cladophora sp. as the most strong, Cyanophyta is secondly, and the snail then is most poor. For the maximum tolerance of the salinity, the snail is about 4.375?, Cyanophyta is about 5.0?, Cladophora sp is then about 7.0?; Under the different salinity for the tolerance , the Cladophora sp. still the most strong, Cyanophyta is secondly, and the snail then is most poor. Under the dark environment, the speed of Cyanophyta begin to be bleaching is very fast than the Cladophora sp.. In the tolerance of pH value range: The snail is about pH=5~10, Cyanophyta is about pH=7~8, Cladophora sp. is about pH=6~8; When the pH value range is in the pH=5~8, the speed of the Cyanophyta occur changing is very fast than Cladophora sp.. The algae are growing on snail shell very different between two ponds, the main reason is water pH value dissimilarly: When pH value over than 8.5, there is no Cladophora sp. to grow on the snail shell, after the pH value to decrease, Cyanophyta then will compare early than Cladophora sp. to grow on the snail shell. Calculate by the classification of the freshwater biological incator : Two organic pollution degree of the ponds may be lain in theβ-mesosaprobic to theα-mesosaprobic, and the polluting degree of the D pond is more seriously. As for two ponds, have already faced what level of eutrophication? Belong to actually which stage of pollution grade? Not only added the classification data of floating and fixative algea in two ponds, and also according to the parts of chemistry analysis method measure of the data makes the substantial evidence, then could carry out the more accurate and thorough study in the days to come steadily studying process.本研究是以臺北市內湖區兩個綠地公園(B公園與D公園)池塘內的石田螺(Sinotaia quadrata)為研究對象,進行以下研究目的之探討:1.鑑別石田螺螺殼上藻類的種類及其與石田螺的互動關係;2.溫度、鹽度、酸鹼值及黑暗等環境因子的差異,對石田螺及螺殼上附生藻類的影響;3.影響藻類附生於石田螺螺殼上的因素;4.將石田螺及螺殼上附生藻類的變化作為監測環境因子或水質變異的指標現象。結果顯示:附生於石田螺螺殼上的藻類有藍綠藻(Cyanophyta)與剛毛藻(Cladophora sp.)兩類;與石田螺的互動關係應屬於片利共生(communalism),但在缺乏食物的情況下,石田螺則會採食同伴殼上的剛毛藻。溫度適應方面,石田螺適宜生存的溫度上限應低於28℃,超過28℃水溫環境的耐受程度,是以剛毛藻為最強,其次是藍綠藻,而石田螺則為最差。對於環境鹽度最大耐受度方面:石田螺約為4.375??,藍綠藻約為5.0??,剛毛藻則約為7.0?;在不同鹽度環境下,鹽度的耐受程度,仍以剛毛藻為最強,其次是藍綠藻,而石田螺則是最差。在黑暗環境下,藍綠藻褪色產生白化現象的速度明顯地比剛毛藻要快了許多。在環境酸鹼值耐受的範圍方面:石田螺約在pH=5~10 之間,藍綠藻約在pH=7~8 之間,剛毛藻則約在pH=6~8 之間;而酸鹼值範圍在pH=5~8 時,藍綠藻產生變化的速度明顯地比剛毛藻還要快。而兩樣區池塘水體酸鹼值的不同,應是造成石田螺螺殼藻類附生現象差異的主要原因:當酸鹼值超過8.5 時,螺殼上就無剛毛藻附生,當酸鹼值降下後,藍綠藻則會比剛毛藻早出現在螺殼上。藉由淡水生物指標的分類推測:兩樣區池塘水體有機污染程度,可能介於β-中腐水性(β-mesosaprobic,βm)至α-中腐水性(α-mesosaprobic,αm)的範圍之間,而D池塘受污染的程度應會比B池塘還要更嚴重些。至於兩樣區池塘水體,已面臨了何種優養化的程度?究竟是屬於哪一個階段的污染等級呢?除須補充水體中浮游性及附著性藻類分類的詳細觀察資料外,仍必須參照部分水質化學分析法所測得的數據作佐證,才能在日後持續地研究過程中進行更精確及深入的探討。
斑馬魚 SULT2 ST2在早期胚胎發育的 RNA 表現
在哺乳動物裡,硫酸化是一種參與外來物解毒作用、內生組織的荷爾蒙調節、藥物代謝及膽汁解毒之重要路徑。其中, SULTZ ( hydroxysteroid sulfotransferase )是能進行上述反應的酵素之一,為了對 SULTZ 的功能與早期發育所扮演的角色作進一步的研究,在本實驗中,我們以班馬魚為模式動物,希望藉由原位雜交法( in situ hybridization )找到 SULTZ 在班馬魚早期胚胎發育的 RNA 表現位置。由目前的實驗結果,發現從卵巢到1-cell、 dome 、 50 %一 epiboly 、 12 小時,都可在胚胎細胞看到訊號表現 · 在 24 小時、 36 小時、 48 小時、 72 小時則可在脊椎兩側體節的肌肉、內胚層、嗅窩、頭部看到訊號表現,此外,在成腦的原位雜交染色結果中亦看到了訊號表現。由此可推論 SULTZ 在斑馬魚早期發育確實扮演了重要的角色。Sulphonation is an important pathway for detoxification of xenobioties, bile acids, drug metabolism, and the regulation of endogenous hormones. SULT2 (hydroxysteroid sulfotransferase) is one of the enzymes which catalyse sulphonation. Zebrafish has emerged as a popular animal model in recent years. Compared with other vertebrates, it provides advantages including ease to get embryos, rapid external development, virtually transparent embryos and ease of genetic manipulation. The above-mentioned strong points made zebrafish a good model animal for us to understand the function of SULT2 during early embryonic development. We performed in situ hybridization to find out the RNA expression of SULT2 during zerbrafish early development. According to our present results, we can detect expression signal on the edge of telencephalon and tectum opticum. the edge on the dorsal zone of corpus cerebelli and ventral zone of periventircular hypothalamus of the adult brain. Besides, we can observe signal evenly distributed in blastocytes of the embryo at 1-cell, dome. 50%-epiboly and 12 hours after fertilization. We also find signal on the muscle next to the spinal cord during the stages of 24, 36, 48 and 72hrs. There are also expressions on hypoblast of embryos at 24, 36 and 72hrs, the olfactory pit at 36 and 4hrs, and strong expression in head region at 48 and 72hrs. These results suggest that SULT2 may have some function at the early development of zebrafish.
萬用虎鉗夾具
機械加工過程中往往遇到形狀複雜工件,無法用一般虎鉗夾持進行加工。若需加工複雜工件時,需使用V 形槽、壓枕……等等夾具加以輔助,但有些夾具根本無法夾持。若用特殊夾具需拆除原有之虎鉗,而且還必須校正,工作繁雜又浪費很多時間。 本設計之優點為不需更換虎鉗,直接放在虎鉗鉗口即可夾持不規則的物體,利用正向力的作用夾持而不打滑,輕易達到夾持時之穩定和足夠之夾持力,以達迅速、不需使用特殊夾具、不需再校正、可當平行塊之多功能夾具,使複雜形狀之工件加工簡單化、迅速化之設計。;When handling workpieces in complicated and irregular shape in the mechanical process, users are unable to make it with ordinary vises. V-block and clamping block might help, while some others do not work at all. In such cases, the user has to tear the vise apart and then do some correction, which is complicated and time-wasting. The strength of this design is that there is no need to replace the vise. The user just puts this device on the vise clamp to clamp the irregular object. The vertical clamping force makes the piece at work stable and allows no slipping. With this device, no special fixture or further correction is needed. It can also be used for a parallel block if necessary. In other words, as a fixture of multiple functions, the device makes the processing work simpler and more efficient than ever.
人面蜘蛛藉調整絲基因表現及網結構促進捕食效率
前人研究顯示結網性蜘蛛會因環境中獵物組成的改變而改變其網結構。然而,除了結構之外,構成網的絲的物理性質與蜘蛛網的捕捉效率也應有密切之關連,但目前並無人探討蜘蛛是否會因獵物而調整絲之化學及物理性質。在本研究中,我以人面蜘蛛為材料,探討:(一)其在面對不同類型之獵物時是否調整其網之結構,絲之胺基酸組成和其物理性質;(二)所產生之改變是否能促進不同類型獵物的捕食效率;以及(三)是獵物之養份或振動刺激引起這些改變。我藉著操控獵物的養份及振動刺激將人面蜘蛛分成四個處理組,分別餵以活蟋蟀(C 組),活蒼蠅(F 組),死蟋蟀加蒼蠅振動刺激(Cd 組)及死蒼蠅加蟋蟀振動刺激(Fd 組)。在完成食控處理後比較各組間之網結構、絲胺基酸組成、絲物理性質、對不同獵物之留置能力以及振動訊號傳遞能力。實驗結果顯示F 組之蜘蛛所結之網有較大的面積及較小的網目,並有較佳之蒼蠅留置力;然而此結構之改變卻會降低其振動訊號傳遞率。C 組之蜘蛛所結之網有較強之張力及較佳之蟋蟀留置力,而且其所產之曳絲有較高比例之alanine、glycine 及glutamine 並較佳之延伸度及韌度,以因應體型較大之獵物。上述結果顯示人面蜘蛛能調整網的結構、絲的蛋白質組成及物理性質來更有效率的捕捉不同類型的獵物。而由於上述各樣變數在C 及F 組間有顯著差異,而在獵物養份與振動訊號錯置的Cd 與Fd 組間無顯著差異,顯示此二項刺激需同時存在才能使人面蜘蛛在面對不同類型獵物時產生結網行為及絲基因表現之改變。Previous studies showed that orb-weaving spiders may adjust web structures when encountering different types of prey. In addition to web structures, silk physical properties should also be critical in affecting the prey catching efficiency of webs. However, so far no one has studied whether spiders adjust silk chemical and physical properties when encountering different types of prey. In this study, I used the giant wood spider Nephila pilipes to investigate (1) would N. pilipes adjust web structures, silk amino acid composition and silk physical properties when encountering different types of prey (2) would such changes enhance the catching efficiency of different prey and (3) were such changes generated by prey chemical composition or vibration signals? To answer these questions, I manipulated prey chemical composition and vibration signals by assigning N. pilipes into the following four feeding groups: (1) living crickets (group C), (2) living flies (group F), (3) dead cricket but with fly vibration signals (group Cd) and (4) dead flies but with cricket vibration signals (group Fd). After the feeding treatments were completed the following variables were compared between the feeding groups: web structures, silk amino acid composition, silk physical properties, retention time of different prey and vibration signal transmission rate of webs. The results showed that the webs built by spiders fed with living flies exhibited larger area, smaller mesh and longer fly retention time. However, such structural changes also reduced the vibration signal transmission ability of the web. On the other hand, webs built by spiders fed with living crickets were stiffer and retained crickets longer. Moreover, their silks exhibited higher percentages of alanine, glycine, glutamine and consequently higher elasticity and strength to cope with the larger prey. These results demonstrated that spiders can adjust web structures, silk protein composition and silk physical properties to enhance the catching efficiency of different prey. While the aforementioned variables differed significantly between spiders in C and F groups, they did not vary between spiders in Cd and Fd groups, in which groups the prey chemical composition and vibration signals were decoupled. Such result suggested that prey chemical composition and vibration signal were both needed in generating changes in web building behavior and silk gene expression in spiders.
無機砷與硒對染色體傷害之交互作用
中文摘要 重金屬對人體傷害的無遠弗屆-人人皆知,根據流行病學的研究指出,長期處在 重金屬含量過高之地區,易造成生理病變。比方說,長期生活在砷含量高的環境中, 易導致肝臟、周邊血管及神經系的損害,各種癌症的發生機率也大為提高。 本實驗以中國倉鼠卵巢細胞(CHO-K1)為材料,利用微核偵測技術(Michronuclei, MN)及姐妹染色分體互換(Sister chromatid exchanges, SCE)之誘引來探討亞硒酸鈉 (Sodium selenite, NaSeO2) 及亞砷酸鈉(Sodium arsenite, NaAsO2) 的交互作用 (interaction)。實驗結果顯示亞硒酸鈉本身不會造成MN 的增加,但讓SCE 頻率增加; 亞砷酸鈉會誘引增加MN 和SCE。亞硒酸鈉前處理不會減少亞砷酸鈉誘引之SCE 和 MN,但可微微降低亞砷酸鈉抑制雙核細胞(Bunucleated cells, BN)的形成,但是不明 顯。本實驗結果未能看出亞硒酸鈉前處理與亞砷酸鈉有明顯的交互作用,但發現亞 硒酸鈉會增加SCE 頻率,因此亞硒酸鈉做為保健食品值得進一步的關切及探討。 英文摘要 It is well-documented that exposure to heavy metals could cause seriously adverse effects to humans. Epidemiological evidence has shown that illness is frequently observed in residents living long-term in heavy metal contaminated area. For example, arsenic exposure was associated with increased incidences of liver diseases, peripheral vascular and neurological diseases, as well as cancers. In this report we investigated the interaction of selenite and arsenite on micronuclei (MN) and sister chromatid exchange (SCE) induction in Chinese hamster ovary-K1 (CHO-K1) cells. The present results demonstrated that sodium selenite by itself did not induce MN, but it did cause the increase of SCE frequency. Sodium arsenite significantly induced MN and SCE in CHO-K1 cells. Pretreatment of CHO-K1 cells with selenite could not reduce MN and SCE frequency induced by arsenite, but slightly prevent the cells from arsenite-induced inhibition of binucleated cell formation. In the present study, no significant interaction between sodium selenite pretreatment and sodium arsenite was observed. However, sodium selenite was unexpectedly found to enhance SCE frequency in CHO-K1 cells. The application of selenite as health protection agent warrants our concerns and further investigation.