【摘要】：研究背景 膽結(jié)石病是消化系統(tǒng)常見(jiàn)疾病,對(duì)人民健康危害極大。膽結(jié)石分為三大類,即膽固醇結(jié)石、棕色膽色素結(jié)石和黑色膽色素結(jié)石。一般認(rèn)為:膽固醇結(jié)石的發(fā)生與膽固醇代謝異常有關(guān),并受膽汁中促成核因子和抑成核因子的共同影響;棕色膽色素結(jié)石由細(xì)菌感染引起,細(xì)菌產(chǎn)生的外源性β-葡萄糖醛酸苷酶水解雙葡萄糖醛酸結(jié)合膽紅素生成游離膽紅素和葡萄糖醛酸,前者與鈣離子結(jié)合形成膽紅素鈣沉淀,同時(shí),細(xì)菌還產(chǎn)生磷脂酶,降解卵磷脂生成棕櫚酸,與Ca~(2+)形成棕櫚酸鈣沉淀,導(dǎo)致棕色膽石形成;黑色膽結(jié)石,呈黑色不規(guī)則外形,質(zhì)地堅(jiān)硬,主要由膽紅素鹽和粘蛋白構(gòu)成,病因不明,多發(fā)生于溶血性疾病和肝硬化患者的膽囊內(nèi)。 芬蘭科學(xué)家Kaiander等進(jìn)行哺乳動(dòng)物細(xì)胞培養(yǎng)時(shí)發(fā)現(xiàn)細(xì)胞內(nèi)存在一種超微生物,能通過(guò)100nm的濾菌器,根據(jù)其體型微小、棲息在血液中的特點(diǎn),將其命名為Nanobacterium sanguineum,簡(jiǎn)稱Nanobacteria,中文譯名納米細(xì)菌。納米細(xì)菌是一種非常古老的超微細(xì)菌,不能利用葡萄糖,主要靠攝取氨基酸來(lái)提供能量,新陳代謝率極其緩慢,約為普通細(xì)菌的1/10,000,當(dāng)生存環(huán)境不利時(shí),能利用環(huán)境中的鈣、磷合成羥基碳酸磷灰石將自身包裹,形成一層厚厚的生物被膜,使其渡過(guò)困難時(shí)期。納米細(xì)菌主要寄生于哺乳動(dòng)物的血液中,由于能透過(guò)0.1um濾菌膜,因此在80%的商用牛血清中存在納米細(xì)菌污染,是導(dǎo)致培養(yǎng)細(xì)胞不明原因死亡的重要因素。普通的細(xì)菌學(xué)方法很難發(fā)現(xiàn)納米細(xì)菌的存在,相差顯微鏡和電子顯微鏡可以清楚地對(duì)其進(jìn)行觀測(cè)。大量納米細(xì)菌通過(guò)受體介導(dǎo)的胞飲作用侵入培養(yǎng)細(xì)胞后,被不完全消化,釋放出內(nèi)毒素,導(dǎo)致受感染細(xì)胞凋亡。研究表明納米細(xì)菌與腎結(jié)石、膽結(jié)石、前列腺炎、多囊腎、多囊肝、動(dòng)脈粥樣硬化等疾病的發(fā)生有關(guān)。研究證實(shí),納米細(xì)菌感染可以導(dǎo)致腎結(jié)石的發(fā)生:另外,膽囊結(jié)石患者血液中納米細(xì)菌的陽(yáng)性率是非結(jié)石患者的4倍,而膽囊結(jié)石患者膽汁納米細(xì)菌感染陽(yáng)性率則高達(dá)61.3%,提示膽汁納米細(xì)菌感染可能是誘發(fā)膽囊結(jié)石的重要原因,因此我們?cè)O(shè)計(jì)本實(shí)驗(yàn),觀察膽汁納米細(xì)菌感染對(duì)實(shí)驗(yàn)兔膽囊結(jié)石的誘發(fā)作用。我們將實(shí)驗(yàn)分兩部分進(jìn)行:第一部分,從膽囊結(jié)石患者膽汁中培養(yǎng)納米細(xì)菌,對(duì)其微生物學(xué)特性進(jìn)行觀察,按文獻(xiàn)方法,對(duì)其存在的真實(shí)性進(jìn)行鑒定:第二部分,將由人膽汁中分離、培養(yǎng)的納米細(xì)菌注入實(shí)驗(yàn)兔的膽囊中,兩周后觀察動(dòng)物成石情況,并對(duì)動(dòng)物的膽囊、膽汁、膽石進(jìn)行分析。
[Abstract]:Background gallstone disease is a common disease of digestive system, which is harmful to people's health. Gallstones are divided into three categories: cholesterol stone, brown pigment stone and black pigment stone. It is generally believed that the occurrence of cholesterol stones is related to the abnormal metabolism of cholesterol, and is affected by both nuclear factors and inhibitory factors in bile. Brown gallstone was caused by bacterial infection. The bacteria produced exogenous 尾 -glucuronidase to hydrolyze diglucuronic acid-conjugated bilirubin to form free bilirubin and glucuronic acid, the former combined with calcium to form calcium bilirubin precipitate. At the same time, bacteria also produced phospholipase, degradation of lecithin to palmitic acid, and Ca~ (2) to form calcium palmitate precipitation, resulting in brown gallstone formation; Black gallstones, with irregular black appearance and hard texture, are mainly composed of bilirubin salt and mucin. The etiology is unknown and occurs in the gallbladder of patients with hemolytic diseases and cirrhosis. In mammalian cell culture, Finnish scientist Kaiander et al. found that there is a kind of ultramicro organism in the cell, which can be called Nanobacteria, by the filter of 100nm, according to its tiny size and perched in blood. Nanometer bacteria. The nanobacteria are very old, ultramicro bacteria that don't use glucose, they're mainly powered by amino acids, and their metabolism is extremely slow, about a tenth of a percent of the normal bacteria, when the environment is bad. It can make use of calcium and phosphorus to synthesize hydroxycarbonate apatite and form a thick biofilm to make it through difficult period. Nanobacteria are mainly parasitic in mammalian blood. Because they can pass through the 0.1um filter membrane, there is contamination of nanobacteria in 80% commercial bovine serum, which is an important factor to cause the unexplained death of cultured cells. It is difficult to find nanobacteria by ordinary bacteriological methods and can be clearly observed by phase contrast microscope and electron microscope. A large number of nanobacteria invaded the cultured cells through receptor-mediated cytosolic action and were incompletely digested and released endotoxin which led to the apoptosis of infected cells. Studies have shown that nano-bacteria are associated with kidney stones, gallstones, prostatitis, polycystic kidney, polycystic liver, atherosclerosis and other diseases. Studies have shown that nanobacterial infections can lead to kidney stones: in addition, the positive rate of nanobacteria in the blood of patients with cholecystolithiasis is four times as high as that of non-stone patients. The positive rate of bile nanobacterial infection in gallstone patients is as high as 61.3, which suggests that bile nanobacterial infection may be an important cause of inducing gallstone, so we designed this experiment. To observe the inducing effect of bile nanobacterial infection on gallstone in rabbits. We divided the experiment into two parts: in the first part, nanobacteria were cultured from bile of cholecystolithiasis patients, their microbiological characteristics were observed, and the authenticity of their existence was identified according to the method of literature. The nanobacteria isolated from human bile were injected into the gallbladder of experimental rabbits. After two weeks, the animal stone formation was observed and the gallbladder, bile and cholelithiasis of animals were analyzed.
【學(xué)位授予單位】：汕頭大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2005
【分類號(hào)】：R363

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