本文選題：骨關(guān)節(jié)結(jié)核 + 維生素D�。� 參考：《南方醫(yī)科大學(xué)》2017年碩士論文

【摘要】：一、研究背景近年來有研究發(fā)現(xiàn),除環(huán)境與生活方式等外界因素外,個(gè)體的遺傳背景是結(jié)核病發(fā)生的另一種影響因素。目前已經(jīng)發(fā)現(xiàn)多個(gè)基因位點(diǎn)與結(jié)核病發(fā)病密切相關(guān),其中維生素D受體(vitaminDreceptor,VDR)是其中主要的候選基因之一。迄今為止,已發(fā)現(xiàn)不少于25個(gè)VDR基因多態(tài)性位點(diǎn),其中研究較多的包括ApaⅠ、Bsm Ⅰ、FokⅠ、TaqⅠ四個(gè)位點(diǎn),是調(diào)節(jié)骨代謝的主要位點(diǎn)。此外,許多VDR目的基因是通過一個(gè)保守的內(nèi)分泌途徑來調(diào)節(jié)維生素D的代謝,所以VDR蛋白分子或其功能上的任何變化都可能影響維生素D的代謝,從而影響骨關(guān)節(jié)結(jié)核的易感性。目前有關(guān)VDR基因多態(tài)性對(duì)于肺結(jié)核易感性的研究在國內(nèi)外已有較多報(bào)道,但VDR基因多態(tài)性對(duì)骨關(guān)節(jié)結(jié)核易感性是否有影響,國內(nèi)外相關(guān)性研究較少。至今尚無針對(duì)廣東地區(qū)漢族人群維生素D受體基因多態(tài)性與骨關(guān)節(jié)結(jié)核易感性的研究。同時(shí),作為VDR的配體,維生素D的血清濃度是否影響骨關(guān)節(jié)結(jié)核的易感性,目前尚不明確。二、研究目的1、通過檢測骨關(guān)節(jié)結(jié)核病人與非骨關(guān)節(jié)結(jié)核者的維生素D受體基因多態(tài)性,探討維生素D受體基因多態(tài)性與中國廣東地區(qū)漢族人群骨關(guān)節(jié)結(jié)核易感性的關(guān)系。2、通過檢測骨關(guān)節(jié)結(jié)核病人與非骨關(guān)節(jié)結(jié)核者的維生素D血清濃度,探討維生素D血清濃度與中國廣東地區(qū)漢族人群骨關(guān)節(jié)結(jié)核易感性的相關(guān)性。三、材料與方法1、研究對(duì)象與分組1.1結(jié)核組收集2015年5月至2016年12月南方醫(yī)院、廣州市胸科醫(yī)院、廣州華僑醫(yī)院、廣州軍區(qū)總醫(yī)院、廣州中醫(yī)藥大學(xué)第一附屬醫(yī)院、河源市中醫(yī)院收治的廣東地區(qū)漢族罹患骨關(guān)節(jié)結(jié)核患者133例,男85例,女48例,年齡8-84歲,平均45.5歲。術(shù)前血液化驗(yàn)指標(biāo)及放射影像學(xué)符合骨關(guān)節(jié)結(jié)核的診斷,術(shù)后病灶病理切片證實(shí)結(jié)核病感染征象,病灶組織分離培養(yǎng)結(jié)核分枝桿菌陽性或抗酸染色陽性,納入結(jié)核組。1.2對(duì)照組2015年5月至2016年12月,選取與病例組無血緣關(guān)系的130例廣東地區(qū)漢族志愿者作為對(duì)照。入選者均來自河源市中醫(yī)院健康查體者;年齡20-76歲,平均43.4歲;其中男性68例,女性62例。均經(jīng)問病史查體及胸片檢查均排除肺結(jié)核及骨關(guān)節(jié)結(jié)核病史。2、樣本采集兩組研究對(duì)象在次日凌晨空腹抽取外周靜脈血2ml,取1ml置于EDTA抗凝管中,靜置在室溫(20-25℃C)1小時(shí)后,2000轉(zhuǎn)/分離心10分鐘,得到的血清置于-80℃C冰箱凍存待檢。剩余1ml,提取VDR全基因組DNA,置于-80℃C冰箱待檢。3、維生素 D 多態(tài)位點(diǎn) rs7975232(aⅠ)、rs1544410(Bsm Ⅰ)、rs2228570(FokⅠ)、rs731236(TaqⅠ)的檢測應(yīng)用SNaPshot技術(shù)檢測兩組對(duì)象維生素D受體基因各位點(diǎn)多態(tài)性,并進(jìn)行VDR基因分型。4、血清維生素D的測定檢測血清25-羥維生素D(250HD)是目前評(píng)價(jià)血清維生素D狀態(tài)的最佳方法。應(yīng)用雙抗體夾心法(Elisa)測定標(biāo)本中人25-羥維生素D水平,評(píng)價(jià)人體血清維生素D的濃度。5、數(shù)據(jù)分析采用SPSS軟件(version 20.0;SPSS,Chicago,IL)進(jìn)行統(tǒng)計(jì)學(xué)分析。計(jì)量資料以均數(shù)±標(biāo)準(zhǔn)差(x±s)表示(本研究的計(jì)量資料符合正態(tài)分布),計(jì)數(shù)資料用頻數(shù)表示。采用卡方檢驗(yàn)比較分類變量及計(jì)數(shù)資料;采用獨(dú)立樣本t檢驗(yàn)比較連續(xù)變量。雙側(cè)P0.05表示差異有統(tǒng)計(jì)學(xué)意義。四、結(jié)果1、Hardy-Weinberg遺傳平衡檢驗(yàn)根據(jù)Hardy-Weinberg遺傳平衡定律計(jì)算對(duì)照組各基因型的預(yù)期值。經(jīng)卡方檢驗(yàn)各觀察值與期望值之間分布近似,P值均0.05,提示對(duì)照組樣本來自于遺傳平衡群體,有較好代表性。2、一般情況比較兩組研究對(duì)象在年齡、性別構(gòu)成方面的差異無統(tǒng)計(jì)學(xué)意義(P0.05)。3、兩組VDR基因型分布頻率比較維生素D受體基因Apa Ⅰ位點(diǎn)的基因型分別為野生型AA和突變型Aa、aa,三種基因型在結(jié)核組和對(duì)照組中的分布差異有統(tǒng)計(jì)學(xué)意義(P0.05);其中結(jié)核組中AA基因型的分布頻率為49.9%,顯著高于對(duì)照組(33.1%),與Aa基因型相比,罹患骨關(guān)節(jié)結(jié)核風(fēng)險(xiǎn)更大(OR=1.924,P＜0.05,95%CI為1.141～3.245)。維生素D受體基因Bsm Ⅰ位點(diǎn)的基因型分別為野生型BB和突變型Bb、bb,但在結(jié)核組和對(duì)照組內(nèi)未能測到突變型bb。兩個(gè)基因型在兩組內(nèi)的分布差異無統(tǒng)計(jì)學(xué)意義(P0.05)。維生素D受體基因Fok Ⅰ位點(diǎn)的基因型分別為野生型FF和突變型Ff、f、,三種基因型在兩組中的分布差異有統(tǒng)計(jì)學(xué)意義(P0.05);其中結(jié)核組中ff基因型的分布頻率為22.6%,顯著低于對(duì)照組36.2%,與FF、Ff基因型相比,罹患骨關(guān)節(jié)結(jié)核風(fēng)險(xiǎn)更小(OR=0.458,P0.05,95%CI 為 0.235～0.893;OR=0.549,P0.05,95%CI 為 0.306～0.982)。維生素D受體基因Taq Ⅰ位點(diǎn)的基因型分別為野生型TT和突變型Tt、tt,但在結(jié)核組和對(duì)照組內(nèi)未能測到突變型tt。兩個(gè)基因型在兩組內(nèi)的分布差異無統(tǒng)計(jì)學(xué)意義(P0.05)。4、兩組血清維生素D濃度比較結(jié)核組與正常對(duì)照組的250HD濃度均數(shù)分別為25.72±6.21、27.27±5.53ng/mL,兩組濃度的差異有統(tǒng)計(jì)學(xué)意義(P0.05)。五、結(jié)論1、維生素D受體基因多態(tài)性位點(diǎn)rs7975232(ApaⅠ)、rs2228570(FokⅠ)的基因型在兩組間分布頻率存在差異,提示其多態(tài)性可能與廣東地區(qū)漢族人群骨關(guān)節(jié)結(jié)核相關(guān);其中ApaⅠ-AA基因型可能是骨關(guān)節(jié)結(jié)核的易感基因型,而Fok Ⅰ-ff基因型可能是骨關(guān)節(jié)結(jié)核的保護(hù)基因型。rs1544410(Bsm Ⅰ)、rs731236(TaqⅠ)基因型分布在兩組間不存在差異,提示其多態(tài)性可能與廣東地區(qū)漢族人群骨關(guān)節(jié)結(jié)核無相關(guān)性。2、結(jié)核組患者血清25-羥維生素D濃度較正常組低,提示血清25-羥維生素D濃度可能與廣東地區(qū)漢族人群骨關(guān)節(jié)結(jié)核發(fā)病相關(guān)。
[Abstract]:1. In recent years, studies have found that the genetic background of individuals is another factor affecting the occurrence of tuberculosis in addition to external factors such as environment and lifestyle. At present, multiple gene loci have been found to be closely related to tuberculosis, and vitamin D receptor (vitaminDreceptor, VDR) is one of the major candidate genes. Up to now, no less than 25 VDR gene polymorphisms have been found, and many of them include Apa I, Bsm I, Fok I, and Taq I, which are the main sites for regulating bone metabolism. In addition, many VDR target genes regulate the metabolism of vitamin D through a conservative endocrine pathway, so VDR protein molecules or their functional functions. What changes may affect the metabolism of vitamin D and thus affect the susceptibility of bone and joint tuberculosis. The research on the susceptibility of VDR gene polymorphism to tuberculosis has been reported at home and abroad, but the polymorphism of VDR gene has an impact on the susceptibility to tuberculosis of bone and joint, and there are few studies at home and abroad. So far, there is no reference to Guangdong. Study on the polymorphism of vitamin D receptor gene and susceptibility to joint tuberculosis in the Han population. At the same time, as the ligand of VDR, whether the serum concentration of vitamin D affects the susceptibility to bone and joint tuberculosis is not yet clear. Two. Objective 1 to detect the multiple vitamin D receptor genes in people with osteoarticular tuberculosis and non osteoarticular tuberculosis. The relationship between vitamin D receptor gene polymorphism and the susceptibility of bone and joint tuberculosis in Han population in Guangdong, China,.2. The correlation between the serum concentration of vitamin D and the susceptibility of human bone and joint tuberculosis in Guangdong area of China was investigated by detecting the serum concentration of vitamin D in patients with tuberculosis and non osteoarticular tuberculosis. Three, three, Materials and methods 1. The subjects and group 1.1 tuberculosis group were collected from May 2015 to December 2016 to collect 133 cases of bone joint tuberculosis in the Han nationality in Guangdong, Guangzhou Chest Hospital, Guangzhou Overseas Chinese Hos, Guangzhou Overseas Chinese Hos, the First Affiliated Hospital of Guangzhou University of Chinese Medicine, 85 men and 48 women. The age of 8-84 years old, average 45.5 years old. Preoperative blood test indexes and radiological imaging accords with the diagnosis of tuberculosis of bone and joint. The pathological sections of the lesion confirm the signs of tuberculosis infection after operation. The tissue isolation and culture of Mycobacterium tuberculosis positive or positive acid stain positive, included in the tuberculosis group.1.2 control group from May 2015 to December 2016, and there is no blood in the case group. 130 cases of Han volunteers in Guangdong area were selected as control. All the participants were selected from the health checkup in Heyuan Hospital of Traditional Chinese Medicine; the age 20-76 years old, with an average of 43.4 years old, including 68 males and 62 women, all excluded tuberculosis and the history of tuberculosis of bone and joint.2 through the examination of medical history and chest examination. The samples were collected in the morning empty in the morning of the next day. Abdominal vein blood 2ml was extracted and 1ml was placed in EDTA anticoagulant tube. After 1 hours at room temperature (20-25 C C), 2000 turns / separated heart 10 minutes. The obtained serum was placed at -80 C C refrigerator to be frozen to be checked. The remaining 1ml was extracted from VDR whole genome DNA, at -80 C C refrigerator to be tested.3, vitamin I polymorphic loci (Fok I), rs731236 (Taq I) detection and application of SNaPshot technology to detect the polymorphism of two groups of vitamin D receptor genes and VDR gene typing.4. Serum vitamin D determination and detection of serum 25- hydroxyvitamin D (250HD) is the best method to evaluate the status of vitamin D in serum. The level of human 25- hydroxyvitamin D was used to evaluate the concentration of vitamin D in human serum. The data analysis was analyzed by SPSS software (version 20; SPSS, Chicago, IL). The measurement data were expressed in mean + standard deviation (x +) (x +.), and the count data were expressed in frequency. The classification variables were compared by chi square test. And counting data, using independent sample t test to compare continuous variables. Bilateral P0.05 showed statistical significance. Four, results 1, Hardy-Weinberg genetic balance test calculated the expected values of each genotype in the control group according to the Hardy-Weinberg genetic balance law. The distribution of each observation value and the expected value was approximately 0.05, and the value of P was 0.05, The sample of the control group came from the genetic balance group and had a better representative.2. In general, there was no significant difference between the two groups of subjects in age and the gender composition (P0.05).3. The two groups of VDR genotype frequencies compared the genotype of the vitamin D receptor gene Apa I loci to the wild type AA and the mutant Aa, AA, and three bases, respectively. The distribution of the type in the tuberculosis group and the control group was statistically significant (P0.05); the frequency of AA genotype in the tuberculosis group was 49.9%, significantly higher than that of the control group (33.1%). Compared with the Aa genotype, the risk of osteoarticular tuberculosis was greater (OR=1.924, P < 0.05,95%CI 1.141 to 3.245). The genotype of the vitamin D receptor gene Bsm I loci Wild type BB and mutant Bb, BB, but there was no significant difference in the distribution of two genotypes of mutant bb. in the two groups (P0.05) in the tuberculosis group and the control group. The genotype of the vitamin D receptor gene Fok I loci were the wild type FF and the mutant Ff, F, and the differences in the distribution of the three genotypes in the two groups were statistically significant Significance (P0.05); the distribution frequency of FF genotypes in tuberculosis group was 22.6%, significantly lower than that of control group 36.2%. Compared with FF and Ff genotype, the risk of osteoarticular tuberculosis was less (OR=0.458, P0.05,95%CI was 0.235 ~ 0.893; OR=0.549, P0.05,95%CI was 0.306 to 0.982). The genotype of the Taq I loci of the vitamin D receptor gene was wild-type TT. And mutant Tt, TT, but in the tuberculosis group and the control group, the distribution difference of the mutant tt. two genotypes in the two groups was not statistically significant (P0.05).4. The concentration of serum vitamin D in the two groups compared with the normal control group was 25.72 + 6.21,27.27 + 5.53ng/mL, respectively, and the difference between the two groups was statistically significant P0.05. (P0.05). Five, conclusion 1, vitamin D receptor gene polymorphism site rs7975232 (Apa I), rs2228570 (Fok I) genotype distribution frequency differences between the two groups, suggesting that the polymorphism may be associated with bone joint tuberculosis in Guangdong Han population, and Apa I -AA genotypes may be the susceptible genotype of bone joint tuberculosis, and Fok I -ff. The genotype may be the protective genotype.Rs1544410 (Bsm I) of bone joint tuberculosis. The genotype distribution of rs731236 (Taq I) is not different between the two groups, suggesting that the polymorphism may be not related to the.2 of the bone and joint tuberculosis in the Han population of Guangdong, and the serum 25- hydroxyvitamine D concentration is lower than that of the normal group, suggesting the serum 25- hydroxyvitamin D. The concentration may be related to the incidence of osteoarticular tuberculosis in Han nationality in Guangdong area.

【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R529.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 廖祥鵬;張?jiān)隼?張紅紅;朱漢民;周建烈;黃琪仁;汪之頊;王亮;劉忠厚;;維生素D與成年人骨骼健康應(yīng)用指南(2014年標(biāo)準(zhǔn)版)[J];中國骨質(zhì)疏松雜志;2014年09期

2 徐金瑞;納小菲;楊易;;寧夏漢族人群VDR基因4個(gè)SNPs位點(diǎn)基因多態(tài)性與2型糖尿病的關(guān)聯(lián)性分析[J];吉林大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2012年05期

3 王黎霞;成詩明;陳明亭;趙雁林;張慧;姜世聞;何廣學(xué);呂青;杜昕;陳偉;劉小秋;阮云洲;王勝芬;夏aa;于蘭;李峻;李雪;;2010年全國第五次結(jié)核病流行病學(xué)抽樣調(diào)查報(bào)告[J];中國防癆雜志;2012年08期

4 唐煒;陳小松;沈坤煒;;維生素D與乳腺癌發(fā)病風(fēng)險(xiǎn)及預(yù)后的關(guān)系[J];外科理論與實(shí)踐;2012年03期

5 李衛(wèi)國;李宇寧;;維生素D缺乏和人類疾病[J];國際兒科學(xué)雜志;2011年05期

6 牟金金;楊敏;柳汝明;周俊翔;唐堯;;維生素D輔助治療肺結(jié)核的系統(tǒng)評(píng)價(jià)[J];中國藥房;2011年28期

7 李曉雙;董克習(xí);邵作峰;李宣東;王芳;;魯南地區(qū)煤礦工人血清25-OH-D水平檢測及相關(guān)因素分析[J];山東醫(yī)藥;2010年35期

8 王曉云;;維生素D受體基因與結(jié)核病[J];中國現(xiàn)代醫(yī)生;2008年23期

9 馬志明;肖唐;林國;劉健雄;譚耀駒;王云南;鄺浩斌;譚守勇;;白介素-10基因多態(tài)性與肺結(jié)核病易感性的研究[J];廣東醫(yī)學(xué);2007年08期

10 葉世輝;劉孟黎;賀晨;劉晟;何海妮;;HLA-DRB1~*等位基因與肺結(jié)核的相關(guān)性研究[J];中國輸血雜志;2007年01期

，

本文編號(hào)：1849229