Ethidium Bromide 對p53 基因缺陷老鼠致癌影響
p53 蛋白是一種轉錄調控因子,其在抑制癌症的形成扮演很重要的角色。已知的報告顯示它能影響下游特定蛋白表現進而調控細胞週期或促使細胞進行凋亡作用。如此作用機制對於可能癌化的細胞在生物體造成傷害之前能被先行消滅,因此正常的p53 蛋白存在與否,對癌症的產生有很大的影響。 根據上述結果,我們想分析不同表現量的p53 是否對於致癌藥劑-EtBr所引發的癌化細胞有所影響。所以我們設計實驗,將具有正常表現量p53 蛋白的老鼠(p53+/+)與僅能表現少量p53 蛋白的老鼠(p53+/-),均塗以等劑量之致癌藥劑EtBr於其大腿皮膚上,觀察其致癌情形的差異。實驗結果發現 : 在塗EtBr 的14 隻p53+/-老鼠中,有3 隻出現了組織病變的症狀,包括脾臟腫大,淋巴組織長出腫瘤,而其他對照組(一)塗EtBr 的p53+/+老鼠,(二)不塗EtBr 的p53+/+老鼠或(三)不塗EtBr 的p53+/-老鼠都沒有產生異常的症狀。因此根據上述實驗結果進行推論,發現經EtBr 的刺激後,p53+/-的老鼠的確較其他的老鼠更容易得到癌化細胞。;p53 is a transcription factor that plays an important role in suppression of tumorgenesis. Previous reports showed it regulates cell cycle and enhances cell apoptosis by effecting specific proteins expression. By this way, the problematic tumor cell can be deleted to protect organism from tumor damage. According above conclusion, we are interested to know whether p53 plays a role involved the carcinogens-EtBr induced tumorgenesis. We compared the differences of tumorgenesis between mice express normal (p53+/+) or low (p53+/-) expression level of p53 proteins when they are smeared equal amount of EtBr on skin of thigh. In here, our results show three of fourteen p53+/- mice had pathological changes, containing spleen enlargement and lymphoma. However, other three comparisons, smear p53+/+ mice with EtBr (1), without EtBr (2) or p53+/- without EtBr(3), are normal. Therefore, We suggest that p53+/- mice are easily to obtain tumor than p53+/+ mice after EtBr treatment.
酒精對斑馬魚胚胎眼睛發育的影響
長期以來,臨床研究發現會影響人類胚胎的正常發育,造成胚胎畸形、成長遲緩等現象,稱為胎兒酒精症候群(fetal alcohol syndrome),但是其病理機制仍不清楚。最近的研究發現斑馬魚胚胎對於酒精有非常靈敏的反應,出現明顯的發育異常現象,例如心臟膨大、眼睛縮小、骨頭變形等現象,與人類胎兒酒精症候群相似,顯示斑馬魚非常適合作為研究胎兒酒精症候群的模式動物。在本實驗中,我們針對酒精影響視網膜神經發育的現象進行探討,並且也利用基因晶片篩選出胚胎發育時期受酒精影的的基因轉錄子,藉以了解酒雞造成胚胎畸形發育的機制。實驗結果顯示在眼睛發育過程中,1.5% 酒精浸泡會抑制視網膜神經細胞的分層(Lamination),而進一步利用RNA定位雜交的方式以眼睛的標示基因rx1、pax6b、six3b、alpha-crystallin、rho、neuroD、huC觀察他們的表現,發現酒精會減少視網膜神經細胞的分化,造成眼睛發育的缺陷。在基因晶片的分析中,我們發現酒精處理會抑制許多控制眼睛發育的基因調控子的表現,包括了crx、rybp、irx4a、optx2、rx1、brn3b1 與 vsx1等,此外約有16%的眼睛相關基因液受到酒精的抑制,遠高於其他基因受抑制的比例,顯示眼睛是酒精傷害最嚴重的區域之一。我們從實驗結果得知酒精並未對胚胎發育初期眼睛的發育造成明顯的影響,但是隨後由於控制細胞分化的幾個重要基因受到酒精抑制作用,造成視網膜神經細胞的分化停止,引起眼睛的異常發育的情況。It was known that prenatal alcohol exposure may cause serious birth defects and developmental disabilities. The molecular mechanism of this fetal alcohol syndrome still remains unclear. Here we used zebrafish embryo as a model to investigate the toxic effect of alcohol in retinal development. The histochemical analysis revealed that the cell lamination was prohibited by alcohol incubation. It appears that the retinal cell differentiation was inhibited. As revealed by whole mount RNA in situ hybridization, it appears that the transcription of a number of retinal-related regulatory genes, including rx1, pax6b, six3b, alpha-crystallin, rho, neuroD and huC, were all inhibited in zebrafish embryo by alcohol exposure. The transcriptional profile of alcohol-exposed embryos was also compared with normal embryos by microarray analysis at different stages. It appears that 16% of retinal-related genes were all repressed by 1.5% alcohol incubation, including several retina-related transcriptional factors, including crx, rybp, irx4a, optx2, rx1, brn3b1and vsx1. Our results suggest that alcohol did not interfere the early development of the eye, but has inhibited the final cell differentiation of retina cells. This study helps us understand the molecular mechanism of alcohol-mediated retinal malformation.
解開蔗糖水解的秘密
本研究利用偏振片、量角器為刻度盤、雷射光為光源,及照度計為偵測器,組裝一個簡易且可靠的旋光度計。我們利用單位時間旋光度的變化量當作反應速率,來測量蔗糖的水解速率,同時求出蔗糖水解反應的反應級數、速率常數(k)。利用糖類的旋光度具有加成性之特性,找出不同混合比例時的旋光度,追蹤實際蔗糖水解的每個狀態,找出最後平衡狀態,同時將蔗糖水解平衡結果顯示,旋光度與濃度有線性關係,而蔗糖水解反應對蔗糖而言為一級反應。接著,我們在蔗糖水溶液中加入不同種類的酸,探討催化劑的種類與蔗糖水解反應速率的關係。 In this research, in order to measure the optical rotation accurately without expensive equipments or complex process, we assembled a polarimeter by ourselves. With simple materials which can be found in ordinary senior high school laboratories, including a calibrated scale, a simple Luxmeter, a laser as the photo source, and other side devices. The Polarimeter ended up operating fluently and accurately. We put the laser under a tube, which has two pieces of polar screens on the top of it and on the bottom of it, ,and put a luxmeter just above the tube. When we slowly rotate the polar screen on the top, the figure shown on the luxmeter changes. By numerical analysis, we can get information about the hydrolysis of polarized substance. Secondary, we measured the optical rotation of glucose, fructose, malt sugar, galactose, and sucrose to get their specific rotation. Then we measured the optical rotation of sucrose every five minutes. By doing this, we could keep track of the hydrolysis rate of sucrose, figure out the order of reaction, and the rate constant (k) and the equilibrium constant (K). Thirdly, we used different kinds of acids into sucrose solution as the catalyst, and observed the effect. The result showed that hydrochloric acid is a better catalyst to this reaction than sulfuric acid and nitric acid. The polarimeter of this research can be used in science education of junior and senior high school. By teaching students to assemble and operate the self-made polarimeter, students can know better about optical rotation and polarized substance. Also, the interest in this experiement will add to students’ motivation to do science research.