長期處理兒茶素對無機砷與氯化鎘之遺傳毒性
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\r 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\r 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;長\r 期處在含量過高的鎘環境中,易引起不正常的鈣代謝﹐產生骨質疏鬆症、軟骨症、\r 糖尿、胺基尿酸、尿蛋白、貧血、和肝功能異常的症狀;而目前坊間所看到各類茶\r 品中含有的兒茶素則被發現具有抗氧化、抗突變及清除自由基等功能。故本實驗以\r 人類胃癌細胞(SC-M1)為材料,以微核(micronuclei)偵測技術觀察細胞長期處理兒茶\r 素((-)-epigallocatechin-3-gallate, EGCG)後,對於亞砷酸鈉(sodium arsenite, NaAsO?)\r 及氯化鎘(cadmium chloride, CdCl?)所造成的細胞遺傳毒性有何影響。另一方面則利\r 用SRB 分析法測量細胞存活率,探討長期處理兒茶素後,亞砷酸鈉及氯化鎘對於細\r 胞的毒殺情況是否改變。實驗結果顯示細胞長期處理兒茶素不會造成微核的增加,\r 但是卻會增加細胞的存活率;單獨處理亞砷酸鈉和氯化鎘則都會使細胞中微核明顯\r 的增加,且會降低細胞的存活率;而長期處理兒茶素的細胞對亞砷酸鈉所誘發的微\r 核有抑制作用且會增加細胞對於亞砷酸鈉的耐受度,長期處理兒茶素無法抑制氯化\r 鎘所誘發的微核與細胞毒性。\r \r \r It is well documented that exposure to heavy metals could cause seriously adverse\r effects to humans. Epidemiological evidence has shown that illness is frequently\r observed in residents living long-term in heavy metal contaminated area. For example,\r long-term exposure of arsenic was associated with increased incidences of liver diseases,\r peripheral vascular and neurological diseases, as well as cancers. And long-term exposed\r to cadmium may lead to abnormal metabolism of calcium, osteoporosis, osteomalacia,\r glucouria, aminoaciduria, proteinuria, anemia and abnormal liver function. Recently,\r numerous reports have shown that (-)-epigallocatechin-3-gallate (EGCG), the major\r polyphenol from green tea, have ability to anti-oxidative stress and anti-mutagenesis. In\r this report, we investigated the effects of EGCG long-term exposure in sodium arsenite\r and cadmium chloride induces chromosomal damages and cytotoxicity in a gastric cancer\r cell line, SC-M1. Our present results demonstrated cells long-term exposure to EGCG did\r not induce micronuclei (MN) formation but induce the growth rate. Sodium arsenite or\r cadmium chloride alone significantly induced MN formation and cytotoxicity in cells\r without long-term EGCG exposure. However, long-term treatment of SC-M1 cells with\r EGCG significantly reduced MN formation and protects the cells from cytotoxicity\r induced by arsenite. Long-term exposure of EGCG had no effect on MN induction and\r cell survival in cadmium-treated SC-M1 cells.
紫茉莉生物時鐘機制的研究
Mirabilis Jalapa is a wild flower, which is often seen in countryside of Taiwan. This plant blossom regularly every day around PM 3:00-6:00. Since its regular behavior, it has the potential to become a model plant for research on circadian ryhtem. According to the on spot observation, a Mirabilis Jalapa will bloom consistently, whereas a minority of flower will bloom earlier or later. First, we observe how Mirabilis Jalapa response to disparate environmental conditions. Under a short period of light(8h) and an average period of light(12), Mirabilis Jalapa acts as normal. But under a long period of light, it flowers approximately 1h ahead of time. When under a continuous light, Mirabilis Jalapa shows a regular flowering in the first four day, and after that, circadian rythem of flowering terminate. The ripe flower buds mentain unbloom for 10 days or more, finally end up withered. Meanwhile, floral meristems are also suppressed. Temperature alter seems affct the flowering slightly. Second, we use SDS-page to detect protein divergence 6h interval.But as many of the earlier attempt, the floral stimuli is not sensible for average protein qualitative analysis. So we decide to take more direct approch by genetic experiment. From supplemental material of article” Orchestrated Transcription of Key Pathways in Arabidopsis by the Circadian” we obtain a group of genes regulared by bioclock in Arabidopsis.With Bio Informatics we choose 41 single gene, and compare with sequences in other species, designing promotors to isolate similar genes in Mirabilis Jalapa. Use rt-PCR to detect the level of certain gene expression,and compare characters of those genes in Mirabilis Jalapa with those genes in Arabidopsis. We also blast those 453 genes to the database of animal, trying to recognize some genes that is widely separated between disparat species. And test these genes in Mirabilis Jalapa to confirm this result.本實驗先針對紫茉莉開花生物時鐘的特性進行觀察,並以不同的變因測試對紫茉莉開花時鐘的影響,紫茉莉個別花朵開花生物時鐘並不一致,長日照會對植株造成開花時間提早(約1 小時)的影響,全日照則會對花芽分生及開花行為造成抑制,溫度對於開花影響不明顯。而後利用蛋白質電泳對不同時間點的樣本進行分析,由於開花激素存量極微,故並電泳結果無明顯差異。再來以阿拉伯芥已知與生物時鐘有關的基因中經過生物資訊(Bio Informatics)預測在紫茉莉中存在與開花及光週期有關之基因(CAA75629 CAB56039 AAB60305 AAC49807 AAA82068),針對該基因進行表現量的測定,驗證該基因與開花生物時鐘的關聯性。針對動物界與植物界中已知與生物時鐘相關的基因進行交叉比對,希望找出跨界廣泛存在的生物時鐘基因,接著測試紫茉莉中該基因的存在與否及表現狀況與其他物種的差異,驗證該基因分布的廣泛性。
漩渦之美
我們常可以在自然界中發現漩渦的存在,但其存在的形體與性質也不盡相同,為了研究漩渦的結構與形體,筆者分析出多種會對漩渦產生影響的因素:開始放流的水而高度、放流洞口大小、有無破壞漩渦結構的阻礙、單孔落流漩渦與雙孔落流漩渦、還有流體的黏滯度對漩渦的影響, 但漩渦是一個不斷改變的流體,非常難以觀察,且自然界的漩渦也不是說出現就出現,所以必須設計一個簡易實驗器材來觀察,並用數位攝影機紀錄下來,再慢慢分析,而我們也可以在這個實驗中了解漩渦的結構,和體會到漩渦所表現出自然界的力與美的一面。‧We can always find in nature of different swirl’s forms and properties. To study the swirls, we analyzed such factors, as the beginning water level, the size of the hole, the presence of obstruction that will destroy the structure of swirls, differences between single-hole-swirls and twin-hole-swirls, and the viscosity of fluid. Because swirls change all the time, it is very difficult to observe. We designed a device .The procedure was recorded with a digital video camera and analyzed it. The study helps us understand the structure of swirls and admire the beauty of swirls.
基因突變對線蟲(Caenorhabdits elegans)之神經系統退化變異株的搜尋以及對其性??
This research is mainly in observation with Caenorhabditis elegans ’s genetic mutation caused via nervous system abnormal character. In the study, I the sample have been cultivated purified and add some chemical material EMS to speed up C.elegans mutation. Then based on the character to further analysis what causeof gene deal with mutation and observe the effects in heredity. The research has two stages, on the first stage of study the mainly target is to both search and purify the mutation of C.elegans. The second stage is based on the exploration of mutation’s searching andpurifying. Because the certain mutation bodies aren’t easy to find out, the project is still on progress at the beginning of second stage, and we conclude some heredity special cases in preliminary of study. 這個實驗主要是觀察並針對線蟲因為基因的突變所產生的神經系統異常的變異性狀,在實驗中我先將樣品線蟲培養並純化至一定數量,並加入適當藥劑EMS造成其突變,經篩選並分析此性狀,進而找出造成其突變之基因,以及觀察此性狀對遺傳表現所造成的影響。 該計畫分成兩階段,第一階段的實驗重點是在突變株的搜尋以及純化上,第二階段則是在突變基因的探討上,由於特定突變株的搜尋並非容易,所以目前計畫只進展至第二階段的遺傳實驗初期,對於其遺傳特徵與突變形式上已有了初步的分析,但尚未定位出該基因的位置。
包埋酵母菌的幾丁球珠對含銅、鐵、鋅、鉛離子之廢水處理及應用
本實驗的目的,就是希望利用幾丁質除污的效果,再配合酵母菌所能累積金屬的能力,以酵母菌包埋於幾丁質的方法,吸附廢水中的重金屬離子。用Langmuir理論求得飽和吸附量,進而求出休眠酵母菌-幾丁質所能吸附金屬離子0.14─0.027(g/g)的數量,與活化酵母菌-幾丁質所能吸附金屬離子1.15─0.050(g/g),並比較回收效率,以應用於處理工業上工廠所排放的廢水。 筆者在偶然的機會將處理過的蟹殼幾丁質固體置於石綿心網上,竟然在數分鐘內像塑膠般熔化,探討原因竟然是石綿心網上的某些金屬鹽所造成,因此筆者希望能透過這發現,用簡單的方法製造出低去乙醯化、高強度及耐中度酸的幾丁基質,並可應用在廢水處理上。;The purpose of this experiment is to explore the effect of using chitin and yeast plat to clarify the heavy metal cation, and that yeast plat works on the accumulation. In the process, the yeast plat is embedded in chitin to absorb the metal cation, therefore, by the theory of Langmuir, the impregnate absorption of dormant yeast-plat-chitin (0.14-0.027[g/g]) can be obtained and yeast-plat-chitin (1.15-0.050[g/g]) can be activated as well. In this way, the efficiency can be compared to apply to work on the effluent in industry. By putting shell-chitin-solid on the wire gouze by chance, we found it melted like plastic in just a few minutes. Some kinds of metal salt on wire gouze cause this change. Therefore, this easy ways can be used to makelow-deamide function group become of high strength and durable under acid environment. The results obtained from this experiment can be applied for the processing of waste liquid.
電源線磁場再生能源的研究與應用
目前正逢能源危機之際,能源再生成為全球關注的課題。有鑒於此,本研究應用高導磁環形鐵粉芯,在表面纏繞多圈漆包線,形成環形管(Toroid)。環形管外圍再繞上交流電源線,電源線內電流產生的磁場,被高導磁環形鐵粉芯所引導,產生較強而均勻的磁場,傳遞至內圈的單心漆包線環形管,依法拉第定律產生電動勢,達到能源再生的目的。藉由六項實驗,驗明我們研究雙環形管理論,推演所得的電動勢公式 ε=(μN1N2/2 r) a2ωIo cosωt 是正確的。再生的電動勢能驅動高亮度的發光二極體提供照明、電器產品充電;還能提供電流過載警示,防止電路過載起火的危險;串、並聯使用則可產出較大功率,深具應用與研究的價值。During mankind are urgent developing of new energy, recycle energy are also one of the global topics; we are using single-heart-Turn around how the high permeability enameled wire ring formation of iron powder core ring solenoid, from the external power supply line also used around - Ring solenoid (Toroid), due to changes current power supply lines of magnetic field generated by the high-permeability core Ring guided iron powder, and can produce more uniform magnetic field so that the inner ring of single-heart enameled wire Ring solenoid, according to Faraday's Law electromotive force can be generated to achieve purpose of recycle energy, we will be divided into six experimental studies to confirm this theory deductions obtained by electromotive forceε=(μN1N2/2 r) a2ωIo cosωt is correct, this electromotive force will enable to supply high brightness LED Optical lighting, can also be used for current overload warning system , if use on series-parallel connection that will produce larger power output, it has great application potential, so the subject is worth to research and development.
N 元二次不定方程式的整數解探討
傳統的畢氏定理三元二次不定方程x² + y² = z²有一組漂亮的整數解為(m² - n²、2mn、m² + n² );中國數學家嚴鎮軍、盛立人所著的從勾股定理談起一書中記載四元二次不定方程x² + y² + z² = w²的整數解為(mn、m² + mn、mn + n²、m²+ mn + n² ),這組解被我們發現有多處遺漏,本文以擴展的畢氏定理做基礎修正了他的整數解公式,並推廣取得N 元二次不定方程的整數解公式。
There is a beautiful integer solution formula for the Pythagorean theorem equation, x² + y² = z² , such as (m² - n² , 2mn ,m² + n² ). The “m" and “n" of the solution formula are integer number. A book written by two Chinese mathematicians, Yen Chen-chun and Sheng Li-jen who expanded the Pythagorean theorem equation to the four variables squares’ indeterminate equation, x² + y² + z² = w² . They claimed that they found its integer solution formula, such as (mn , m² + mn , mn + n² , m² + mn + n² ) for any integer “m" and “n". But we found it losses many solutions. This paper corrected their faults due to the expanded Pythagorean theorem built by ourselves. Further more, we derived a general formula of N variables squares’ indeterminate equation. Now, we can get integer solutions of the equation, (for all natural number “n") easily by choosing integers m1 , m2 , m3 ,……, mn−1 up to you.
人參中人參皂素之分析與食用方式及品種比較
高效液相層析儀(HPLC)為測定中藥指標成分常用的分析工具,而近年來高效液相層析質譜儀(LC-MS)的應用,亦在天然藥物分析上逐漸扮演重要角色。本研究利用HPLC 分析人參中的12 種人參皂素,並以 LC-MS 確認各化合物。
本研究分為三個部份,第一部份為開發適當的HPLC 分析方法分析人參中的主要成份,並使用LC-MS 確認這些化合物。人參中的12 種人參皂素( ginsenosides Rb1, Rb2, Rc, Rd, Re, Rf, Rg1, Rg2, Ro and malonylginsenosides mRb1, mRb2 and mRc ) 可於60 分鐘內成功分離,在 HPLC 分析方法中使用Cosmosil 5C18 管柱,以200nm 偵測,並以20mM KH2PO4-CH3CN 進行梯度沖提, 在LC-MS 分析中則以20mM CH3COONH4-CH3CN 進行梯度沖提。
第二部份為利用上述分析方法,比較同批人參藥材進行不同處理方式後,所得人參皂素含量的多寡,包含泡茶、煮茶、泡酒等。結果顯示若為切段(飲片)泡茶,至少應浸泡30 分鐘,切段(飲片)水煮則以15-20 分鐘較佳,粉末(茶包)泡茶的變化不明顯,而切段酒浸可以得到最多量的人參皂素,且浸泡越久含量越高。
第三部份為使用多變量統計軟體SPSS 進行區別分析(DA),統計結果可作為人參藥材基原辨識之依據。
High-performance liquid chromatography (HPLC) is a common analytical tool that has been used to analyze the index components of Chinese herb medicine. In recent years, high-performance liquid chromatography-mass spectrometry (LC-MS) gradually plays an important role. In this study, we used HPLC to detect 12 ginsenosides in Ginseng Radix, and used LC-MS to identify the 12 compounds.
This study includes three parts. The first part concerns the development of HPLC method for determining the chemical constituents in Ginseng Radix, and using LC-MS to identify them. In Ginseng Radix, components such as ginsenosides Rb1, Rb2, Rc, Rd, Re, Rf, Rg1, Rg2, Ro and malonylginsenosides mRb1, mRb2 and mRc could be separated successfully in 60 mins. They were detected at 200nm by using a Cosmosil 5C18 column with a linear gradient elution of 20mM KH2PO4-CH3CN in the HPLC method, and with a linear gradient elution of 20mM CH3COONH4-CH3CN in the LC-MS method.
In the second part, we used this method mentioned above to evaluate the different results from various processes, which include brewing snippets in hot water, brewing powders in hot water, cooking snippets with boiling water, and brewing snippets in rice wine. Our results are listed as follows, First, the tea using Ginseng Radix snippets brewing for about 30 mins in hot water has the best result. Second, using Ginseng Radix snippets cooking for about 15-20 mins with boiling water has the best result. Third, using Ginseng Radix powders brewing in hot water has no apparent difference. Finally, using Ginseng Radix snippets brewing in the rice wine can get most ginsenosides.
The third part is using the statistic software SPSS to proceed the Discriminate Analysis(DA). The results of the statistic analysis can help us to identify the origin of the herb medicines.
為「世紀之毒」找解藥!- 探討以Pseudomonas mendocina 菌株降解污染土壤中戴奧?
本研究採自中石化安順廠戴奧辛污染場址之土壤,篩選出一株純 菌微生物(Pseudomonas mendocina NSYSU) , 其含有PCDD/Fs (Polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans, PCDD/Fs)土壤於實驗室進行微生物降解試驗,結果顯示約21 天即有 明顯降解結果;本研究同時觀察到該微生物能在汞濃度達50 ppm 之 戴奧辛土壤中,仍具有相當程度之耐受度,並進一步降解戴奧辛化合 物。 我更以標準品探討菌種對於戴奧辛類化合物(Dioxin like congener) 之降解效果,成功地發現這株菌種不僅對於PCDD/Fs 有降 解能力,對於戴奧辛類化合物也有明顯之降解效果。 由研究結果,明顯看出P. mendocina 菌株為「世紀之毒」找到了 解藥,開發出以生物復育改善污染環境的一種新方法。A dioxin-degrading bacterial strain Pseudomonas mendocina NSYSU was isolated from dioxin contaminated soil by selective enrichment techniques. In our previous study, P. mendocina NSYSU was found to be able to use pentachlorophenol (PCP) as its sole carbon source and energy source and was capable of completely degrading this compound. Moreover, P. mendocina NSYSU was also able to mineralize a high concentration of PCP up to 150 mg/L. In this study, P. mendocina NSYSU was investigated for its ability to degrade polychlorinated dibenzofurans (PCDFs) and polychlorinated dibenzo-p-dioxins (PCDDs). Results show that P. mendocina NSYSU could grow well in media containing both PCDFs and PCDDs, and was able to degrade both compounds efficiently. In this study, isotope dilution method and a high resolution gas chromatography-mass spectrometry (HRGC/HRMS) technique were applied for the analyses of PCDFs and PCDDs. Investigation results reveal that significant biodegradation of octachlorinated dibenzo-p-dioxin (OCDD) and octachlorinated dibenzofuran (OCDF) by treating with P. mendocina NSYSU resting cells was observed. The results also indicate that this bacterial strain is able to biodegrade OCDD and OCDF effectively under anoxic conditions due to its facultative anaerobic character. No accumulation of inhibitory toxic byproducts was found in this study. These findings suggest that in situ or on-site bioremediation of dioxin-contaminated soils by using indigenous microorganisms or inoculated P. mendocina NSYSU strain would be a feasible technology for field application.