本文選題：鼠李糖乳桿菌 + 降血脂��； 參考：《揚(yáng)州大學(xué)》2015年博士論文

【摘要】：血清中的血脂水平過(guò)高是引起眾多心血管疾病的主要原因。研究表明,降低血清中的血脂水平能夠顯著降低心血管疾病的死亡率。藥物治療是當(dāng)前治療心血管疾病的主要方式,但是藥物治療成本較高,并且具有較大的副作用。乳酸菌作為人體腸道中重要的益生菌,已被證實(shí)能夠改善血清中的血脂水平,并使腸道微生態(tài)系統(tǒng)發(fā)生長(zhǎng)期而有益的變化,而來(lái)源于人體自身的益生菌通常具有較高的安全性,因此本文從廣西巴馬長(zhǎng)壽地區(qū)的人群腸道中分離篩選出體外輔助降血脂能力較強(qiáng)的乳酸菌,并研究其在高血脂大鼠體內(nèi)的降血脂效果以及對(duì)高血脂大鼠腸道菌群的影響,從而探索乳酸菌在體內(nèi)降血脂的可能機(jī)制,并為以腸道菌群為靶點(diǎn)進(jìn)行的益生菌干預(yù)治療提供理論依據(jù)。1.利用選擇性培養(yǎng)基從廣西巴馬長(zhǎng)壽地區(qū)人群采集的糞便樣品中分離乳酸菌,通過(guò)體外試驗(yàn)篩選出降膽固醇及甘油三酯能力較強(qiáng)的乳酸菌,然后進(jìn)行模擬人工胃液、膽鹽耐受性、抑菌及耐藥性試驗(yàn)；并研究其生長(zhǎng)菌體、熱致死菌體及休眠菌體的膽固醇降解率及其在體外的降膽固醇方式。結(jié)果表明,從15份糞便樣品中分離獲得156株乳酸菌,其中Lp2、S10、Lf7及Hsryfm 1301等16株乳酸菌的膽固醇及甘油三酯的降解率均分別大于30.35%和5.33%, Lp2、S10、Lf7及Hsryfm 1301在pH3.0的人工胃液中存活率均大于63.00%,在含0.1%、0.3%及0.5%膽鹽的培養(yǎng)基中存活率均大于15.00%；Hsryfm 1301的抑菌能力較強(qiáng)于Lp2、S10及Lf7.對(duì)大腸桿菌、沙門氏菌及金黃色葡萄球菌的抑菌圈直徑分別為14.67mm、11.35mm及10.67mm；Hsryfm 1301的耐藥性低于Lp2、S10及Lf7,對(duì)頭孢唑啉、頭孢拉定、四環(huán)素、氯霉素、利福平素均敏感,對(duì)鏈霉素中度敏感,對(duì)青霉素G、諾氟沙星及環(huán)丙沙星均耐藥;Hsryfm 1301在含0.1%、0.3%及0.5%膽鹽的培養(yǎng)基中的降膽固醇能力較強(qiáng)于Lp2、S10及Lf7,降解率分別為56.25%、28.91%及13.20%；Lp2、S10、Lf7及Hsryfm 1301的生長(zhǎng)菌體、熱致死菌體及休眠菌體均能降低培養(yǎng)基中的膽固醇,Hsryfm 1301生長(zhǎng)菌體的膽固醇降解率為61.90%,高于Lp2、S10及Lf7；Hsryfm 1301體外降膽固醇的主要方式為菌體吸收及共沉淀,分別占其總降解率的62.50%及36.50%。經(jīng)API及16S rDNA測(cè)序?qū)sryfm 1301鑒定為鼠李糖乳桿菌(Lactobacillus rhamnosus)。2.通過(guò)飼喂4周高脂飼料來(lái)建立高血脂大鼠模型,以降血脂藥物血脂康作為陽(yáng)性對(duì)照,研究L. rhamnosus hsryfm 1301的菌體懸浮液、熱致死菌體懸浮液、菌體破碎液及其發(fā)酵乳在高血脂大鼠體內(nèi)的降血脂效果。結(jié)果表明,飼喂4周高脂飼料,模型組大鼠血清中的膽固醇(Serum total cholesterol, TC)、甘油三酯(Triglycerides, TG)及低密度脂蛋白(Low density lipoprotein cholesterol, LDL-C)含量顯著高于對(duì)照組(P0.05),分別為2.05mmol/L、1.98mmol/L、0.28mmol/L,表明高血脂大鼠模型建模成功；灌胃4周L. rhamnosus hsryfm 1301發(fā)酵乳后,大鼠對(duì)飼料的利用率顯著高于模型組(P0.05),為29.46%；飼喂8周高脂飼料后,模型組大鼠的肝指數(shù)及體脂指數(shù)均顯著高于對(duì)照組及L. rhamnosus hsryfm 1301各處理組(P0.05),分別為3.98和3.55。灌胃4周L. rhamnosus hsryfm 1301的各處理方式后,菌體懸浮液組及發(fā)酵乳組大鼠血清中TC、TG的含量顯著低于模型組(P0.05),分別為1.50mmol/L、1.30mmol/L及1.53mmol/L、 1.28mmol/L;菌體熱致死組大鼠血清中TC、高密度脂蛋白(High density lipoprotein cholesterol, HDL-C)及LDL-C的含量顯著低于模型組(P0.05),分別為1.90mmol/L、 0.38mmol/L及0.19mmol/L;菌體破碎液組大鼠血清中TC、TG及LDL-C的含量顯著低于模型組(P0.05),分別為1.78mmol/L、1.68mmol/L及0.17mmol/L;灌胃4周的血脂康后,人鼠血清中TC、TG、HDL-C及LDL-C的含量也顯著低于模型組(P0.05),分別為1.76mmol/L、 1.65mmol/L、0.39mmol/L及0.19mmol/L。灌胃4周L. rhamnosus hsryfm 1301的菌體懸浮液、菌體破碲液及其發(fā)酵乳后,大鼠血清中氧化物歧化酶(Superoxide dismutase, SOD)濃度顯著高于模型組(P0.05),分別為74.96U/mL.73.58U/mL及73.96U/mL,而熱致死菌體對(duì)大鼠血清中的SOD濃度影響較��；菌體懸浮液組及發(fā)酵乳組大鼠糞便中膽固醇、甘油三酯和膽汁酸的含量顯著高于模型組(P0.05),分別為10.23μmol/g、4.25μmol/g和28.87μmol/g及9.85μmol/g、4.03pμmol/g和27.13μmol/g;菌體懸浮液組、菌體破碎液組與發(fā)酵乳組大鼠糞便中的水分含量顯著高于模型組(P0.05),分別為46.15%、45-35%及46.85%；通過(guò)透射電鏡觀察發(fā)現(xiàn),L. rhamnosus hsryfm 1301菌體懸浮液、菌體破碎液及其發(fā)酵乳均能預(yù)防由高血脂引起的大鼠肝臟和心臟組織的病變。3.利用聚合酶鏈?zhǔn)椒磻?yīng)一變性梯度凝膠電泳(Polymerase Chain Reaction-Denaturing gradient gelelectrophoresis, PCR-DGGE)技術(shù)研究基礎(chǔ)飼料、高脂飼料及L. rhamnosus hsryfm1301的菌體懸浮液、熱致死菌體懸浮液、菌體破碎液及其發(fā)酵乳對(duì)高血脂大鼠腸道菌群的影響。結(jié)果表明,飼喂4周高脂飼料增加了大鼠腸道中柔嫩梭菌屬PCR-DGGE圖譜的條帶數(shù),降低了腸道微生物16S rRNA基因V3區(qū)、擬桿菌屬、乳酸菌及雙歧桿菌屬PCR-DGGE圖譜的條帶數(shù),而L. rhamnosus hsryfm 1301的各處理方式對(duì)大鼠腸道微生物的影響正好與之相反。飼喂4周基礎(chǔ)飼料及高脂飼料后,大鼠腸道中Klebsiella pneumoniae及Uncultured bacterium的豐度下降甚至消失,同時(shí)增加了大鼠腸道中柔嫩梭菌的種類,分別為Clostridiumbotulinum A str. Hall、Clostridium pasteurianum BC1、Ruminococcus champanellensis 18P13 、Ruminococcus albus 7及Ruminococcus bromii L2-63,而對(duì)照組大鼠腸道中的Uncultured Bacteroidetes bacterium TM1-40肖失。試驗(yàn)第9周,灌胃4周L. rhamnosus hsryfm 1301菌體懸浮液后,大鼠腸道中出現(xiàn)了Pantoea vagans及Clostridium thermocellum,對(duì)照組及模型組大鼠腸道中出現(xiàn)了Streptomyces coelicolor及Clostridiales genomo sp.,對(duì)照組大鼠腸道中Clostridium botulinum A str. Hall的豐度高于其它組；熱致死菌體懸浮液、菌體破碎液及其發(fā)酵乳使大鼠腸道中Ruminococcus bromii L2-63的豐度下降或消失,菌體懸浮液增加了大鼠腸道中Lactobacillus amylovorus GRL的豐度,菌體破碎液增加了大鼠腸道中Lactobacillus buchneri NRRL B-30929的豐度,發(fā)酵乳則增加了大鼠腸道中Lactobacillus sp,的豐度；菌體懸浮液、菌體破碎液及發(fā)酵乳還增加了大鼠腸道中乳酸菌的種類,分別為L(zhǎng)actobacillus murinus DLM 3108、Lactobacillus gasseri ATCC 33323及Lactobacillus sakei subsp.。4.利用實(shí)時(shí)熒光定量聚合酶鏈?zhǔn)椒磻?yīng)(Real-time ploymerase chain reaction, RT-PCR)的方法研究基礎(chǔ)飼料、高脂飼料及L. rhamnosus hsryfm 1301的菌體懸浮液、熱致死菌體懸浮液、菌體破碎液及其發(fā)酵乳對(duì)高血脂大鼠腸道菌群的影響,并分析高血脂大鼠的腸道菌群與其血清血脂水平的相關(guān)性。結(jié)果表明,飼喂8周基礎(chǔ)飼料顯著增加了大鼠腸道中擬桿菌和腸球菌的數(shù)量(P0.05),飼喂8周高脂脂飼料顯著增加了大鼠腸道中柔嫩梭菌、腸球菌以及腸桿菌的數(shù)量(P0.05),同時(shí)顯著降低了擬桿菌、乳酸菌及雙歧桿菌的數(shù)量(P0.05);L. rhamnosus hsryfm 1301各處理方式干預(yù)4周后,菌體懸浮液顯著增加了大鼠腸道中擬桿菌及乳酸菌的數(shù)量(P0.05),并顯著降低了柔嫩梭菌的數(shù)量(P0.05)；熱致死菌體懸浮液顯著增加了大鼠腸道中擬桿菌及腸球菌的數(shù)量(P0.05),并顯著降低了柔嫩梭菌及腸桿菌的數(shù)量(P0.05)；菌體破碎液顯著增加了大鼠腸道中擬桿菌、乳酸菌及雙歧桿菌的數(shù)量(P0.05),并顯著降低了柔嫩梭菌的數(shù)量(P0.05)；發(fā)酵乳顯著增加了大鼠腸道內(nèi)擬桿菌及腸球菌的含量(P0.05),同時(shí)顯著降低了柔嫩梭菌及腸桿菌的數(shù)量(P0.05)；高血脂大鼠腸道內(nèi)柔嫩梭菌的數(shù)量同其血清中TC、TG、HDL-C及LDL-C的含量呈顯著正相關(guān)(P0.05),腸道內(nèi)乳酸菌及雙歧桿菌的數(shù)量同腸桿菌的數(shù)量呈顯著負(fù)相關(guān)(P0.05)。5.利用高通量測(cè)序的方法研究基礎(chǔ)飼料、高脂飼料及L. rhamnosus hsryfm 1301菌體懸浮液及其發(fā)酵乳對(duì)高血脂大鼠腸道菌群的影響,并分析高血脂大鼠的腸道菌群與其血清血脂水平的相關(guān)性。結(jié)果表明,高脂飼料、L. rhamnosus hsryfm 1301菌體懸浮液及其發(fā)酵乳顯著改變了大鼠腸道中柯林斯菌屬、擬桿菌屬、普氏菌屬、乳桿菌屬、腸球菌屬、瘤胃球菌屬等22個(gè)屬的微生物的相對(duì)豐度(P0.05)；飼喂8周基礎(chǔ)飼料及4周高脂飼料降低了大鼠腸道中微生物物種的豐度及多樣性,使得大鼠腸道菌群的Chaol/ACE署Shannon指數(shù)顯著降低(P0.05),Simpson指數(shù)顯著增加(P0.05)；而L. rhamnosus hsryfm 1301菌體懸浮液及其發(fā)酵乳的干預(yù)恢復(fù)了大鼠腸道中微生物物種的豐度及多樣性,顯著增加了大鼠腸道菌群的Chaol/ACE和Shannon旨數(shù)(P0.05),并降低了Simpson旨數(shù)(P0.05)；同時(shí)促進(jìn)了大鼠腸道中乳桿菌屬的繁殖,并抑制了大腸桿菌／志賀氏菌的生長(zhǎng)。大鼠腸道中瘤胃球菌屬的相對(duì)豐度與血清中TG的含量、Dorea spp的相對(duì)豐度與血清中TC及LDL-C的含量、腸球菌屬的相對(duì)豐度與血清中HDL-C的含量均呈顯著正相關(guān)(P0.05),而丁酸弧菌屬的相對(duì)豐度與血清中TC及LDL-C的含量呈顯著負(fù)相關(guān)(P0.05)。
[Abstract]:The high level of blood lipids in the serum is the main cause of many cardiovascular diseases. Studies have shown that reducing blood lipid levels in the serum can significantly reduce the mortality of cardiovascular diseases. Drug treatment is the main treatment of cardiovascular disease at present, but the cost of drug treatment is higher, and it has a larger side effect. As an important probiotic in human intestinal tract, it has been proved to be able to improve blood lipid levels in the serum and to make long-term and beneficial changes in the intestinal microecosystem, while the probiotics derived from the human body often have high safety. In order to explore the effect of blood lipid lowering in hyperlipidemic rats and the effect on the intestinal flora of hyperlipidemia rats, the possible mechanism of lactic acid bacteria in blood lipid reduction in the body was explored, and the theory of probiotic intervention therapy based on the intestinal microflora as the target for the treatment was based on the.1. selection medium. Lactic acid bacteria were isolated from the fecal samples collected from the longevity region of the West Bama, and the cholesterol and triglyceride lactobacillus with strong ability to reduce cholesterol and triglycerides were screened in vitro, and then simulated artificial gastric juice, bile salt tolerance, bacteriostasis and drug resistance test, and the cholesterol degradation rate of the growing mycelium, heat lethal and dormant bodies and the rate of cholesterol degradation were studied. The results showed that 156 strains of lactic acid bacteria were isolated from 15 fecal samples, of which the cholesterol and triglyceride degradation rates of 16 strains of lactic acid bacteria such as Lp2, S10, Lf7 and Hsryfm 1301 were respectively greater than 30.35% and 5.33%, Lp2, S10, Lf7 and Hsryfm 1301 were more than 63% in pH3.0, with 0 in the artificial gastric juice. The survival rates of.1%, 0.3% and 0.5% of bile salts were greater than 15%, Hsryfm 1301 was stronger than Lp2, S10 and Lf7. were 14.67mm, 11.35mm and 10.67mm for Escherichia coli, Salmonella and Staphylococcus aureus, and Hsryfm 1301 was lower than Lp2, S10 and Lf7, and cefazolin, Cefradine, and four rings. It was sensitive to chloramphenicol and rifampicin, moderately sensitive to streptomycin, resistant to penicillin G, norfloxacin and ciprofloxacin, and Hsryfm 1301 in 0.1%, 0.3%, and 0.5% bile salts was stronger than Lp2, S10 and Lf7, and the degradation rates were 56.25%, 28.91% and 13.20%, Lp2, S10, Lf7 and Hsryfm 1301. Both lethal and dormant bacteria could reduce the cholesterol in the medium. The cholesterol degradation rate of Hsryfm 1301 growth bacteria was 61.90%, higher than Lp2, S10 and Lf7. The main way to reduce cholesterol in Hsryfm 1301 was bacterial absorption and co precipitation, accounting for 62.50% of the total degradation rate and 36.50%. by API and 16S rDNA sequencing to identify Hsryfm 1301. A hyperlipidemic rat model was established for Lactobacillus rhamnosus.2. by feeding a high fat diet for 4 weeks. A blood lipid lowering drug xuexuekang was used as a positive control to study the bacterial suspension of L. rhamnosus hsryfm 1301, heat lethal mycelium suspension, bacterial breakage and its fermented milk in hyperlipidemia rats. The results showed that the levels of serum cholesterol (Serum total cholesterol, TC), triglyceride (Triglycerides, TG) and low density lipoprotein (Low density lipoprotein cholesterol, LDL-C) in the model group were significantly higher than those of the control group (P0.05). Rat model was successfully modeled. After 4 weeks of L. rhamnosus hsryfm 1301 fermented milk, the utilization rate of rat feed was significantly higher than that of model group (P0.05), which was 29.46%. After feeding for 8 weeks high fat feed, the liver index and body fat index of the model group were significantly higher than that of the control group and L. rhamnosus hsryfm 1301 treatment group (P0.05), 3.98 and 3.55. respectively. After the treatment of L. rhamnosus hsryfm 1301 for 4 weeks, the content of TC and TG in the serum of the mycelium suspension group and the fermented milk group was significantly lower than that of the model group (P0.05), which were 1.50mmol/L, 1.30mmol/L and 1.53mmol/L, 1.28mmol/L, and the serum of the rats in the body heat death group was TC, and the high-density lipoprotein (High) The content of -C) and LDL-C was significantly lower than that of the model group (P0.05), which were 1.90mmol/L, 0.38mmol/L and 0.19mmol/L, respectively. The content of TC, TG and LDL-C in the serum of the strain group was significantly lower than that of the model group (P0.05). The concentration of oxide dismutase (Superoxide dismutase, SOD) in serum was significantly higher than that in model group (P0.05), 1.76mmol/L, 1.65mmol/L, 0.39mmol/L and 0.19mmol/L. for 4 weeks L. rhamnosus hsryfm 1301, and the concentration of Superoxide dismutase, SOD) in the serum of the rats was significantly higher than that of the model group (P0.05). 6U/mL, and the effect of heat lethal mycelium on the concentration of SOD in the serum of rats was less. The content of cholesterol, triglyceride and bile acid in the feces of the mycelium suspension group and the fermented milk group was significantly higher than that of the model group (P0.05), 10.23 mu mol/g, 4.25 mol/g and 28.87 u mol/g and 9.85 mu mol/g, 4.03p u mol/g and 27.13 micron mol/g, and the bacterial suspension group, The water content in the feces of the mycelial breakage group and the fermented milk group was significantly higher than that of the model group (P0.05), 46.15%, 45-35% and 46.85%, respectively. Through transmission electron microscopy, it was found that the L. rhamnosus hsryfm 1301 mycelium suspension, the mycelial breakage and the fermented milk could prevent the pathological changes of the liver and heart tissues of rats induced by hyperlipidemia.3. Using polymerase chain reaction (Polymerase Chain Reaction-Denaturing gradient gelelectrophoresis, PCR-DGGE) to study the basic feed, high fat feed and L. rhamnosus hsryfm1301 bacterial suspension, thermally lethal mycelium suspension, bacterial breakage and fermented milk for the intestinal flora of hyperlipidemia rats The results showed that 4 weeks of high fat feed increased the number of bands in the PCR-DGGE map of the intestinal tract of the genus Clostridium tenx in rats, and reduced the number of bands of the 16S rRNA gene in the intestinal microbes, the bacillus, Lactobacillus and Bifidobacterium in the PCR-DGGE map of the genus 16S, and the treatment of L. rhamnosus hsryfm 1301 on the intestinal microorganism in rats. On the contrary, the abundances of Klebsiella pneumoniae and Uncultured bacterium in the intestines of rats were reduced or even disappeared after feeding for 4 weeks of basal diet and high fat diet, and the species of Clostridium tenx in the intestines of rats were increased, respectively, Clostridiumbotulinum A str. Hall, Clostridium pasteurianum BC1, Ruminococcus Ruminococcus. Is 18P13, Ruminococcus albus 7 and Ruminococcus bromii L2-63, and Uncultured Bacteroidetes bacterium TM1-40 Shaw loss in the intestinal tract of the control group. After ninth weeks of trial, the intestine of the rat was 4 weeks of L.. Streptomyces coelicolor and Clostridiales genomo sp. were found in the intestinal tract, and the abundance of Clostridium botulinum A str. Hall in the intestinal tract of the control group was higher than that of the other groups; the bacterial suspension and the fermentation milk made the Ruminococcus bromii abundances in the intestine of the rats decreased or disappeared, and the suspension of the bacterial body increased. The abundance of Lactobacillus amylovorus GRL in the intestine of rats increased the abundance of Lactobacillus buchneri NRRL B-30929 in the intestinal tract of the rat, and the fermented milk increased the abundance of Lactobacillus SP in the intestine of the rat, and the bacterial suspension, the mycelium breakage and the fermented milk also increased the species of lactic acid bacteria in the rat intestine. Lactobacillus murinus DLM 3108, Lactobacillus gasseri ATCC 33323 and Lactobacillus sakei subsp..4. use real time fluorescent quantitative polymerase chain reaction (Real-time ploymerase chain) to study base feed, high fat feed and 1301 bacterial suspension, heat lethal bacterial suspension The effect of liquid, mycelium breakage and its fermented milk on the intestinal flora of hyperlipidemia rats and the correlation between the intestinal flora of hyperlipidemia rats and the serum lipid levels were analyzed. The results showed that the number of bacilli and Enterococcus in the intestines of rats was significantly increased by feeding 8 weeks (P0.05), and the rats fed with high fat fat diet for 8 weeks significantly increased the rats. The number of Clostridium tenx, Enterococcus and Enterobacteriaceae (P0.05) significantly decreased the number of bacteriobacteria, lactic acid bacteria and bifidobacteria (P0.05), and the number of bacteriobacteria and lactic acid bacteria (P0.05) in the intestinal tract was significantly increased after 4 weeks of intervention by L. rhamnosus hsryfm 1301, and the tender shuttle was significantly reduced. The number of bacteria (P0.05); the number of bacteriobacteria and Enterococcus (P0.05) in the intestinal tract was significantly increased by the heat lethal bacterial suspension, and the number of Clostridium tenosium and Enterobacter cloacae (P0.05) was significantly reduced. The bacterial breakage significantly increased the number of bacteriobacteria, lactic acid bacteria and bifidobacteria (P0.05) in the intestines of rats (P0.05), and significantly reduced the tender shuttle. The number of bacteria (P0.05); fermented milk significantly increased the content of bacteriobacterium and Enterococcus (P0.05) in the intestines of rats, and significantly decreased the number of Clostridium tenoides and Enterobacteriaceae (P0.05). The number of Clostridium tenosi in the intestinal tract of hyperlipidemia rats was significantly positively correlated with the content of TC, TG, HDL-C and LDL-C in the serum (P0.05), and the lactic acid bacteria in the intestinal tract and the intestinal Lactobacillus in the intestinal tract. The number of bifidobacteria was negatively correlated with the number of Enterobacteriaceae (P0.05).5. using high throughput sequencing method to study the effects of high fat feed and L. rhamnosus hsryfm 1301 suspension and its fermented milk on the intestinal flora of hyperlipidemic rats, and to analyze the phase of intestinal flora and serum lipid levels in hyperlipidemic rats. The results showed that high fat feed, L. rhamnosus hsryfm 1301 mycelium suspension and its fermented milk significantly changed the relative abundance of 22 microbes in the intestinal tract of Collins, pridella, Lactobacillus, Enterococcus and rumen, and 8 weeks of basal feed and 4 weeks of high fat diet were reduced. The abundance and diversity of microbial species in the intestinal tract of rats greatly reduced the Shannon index of Chaol/ACE in the intestinal flora of rats (P0.05), and the Simpson index increased significantly (P0.05), while the intervention of L. rhamnosus hsryfm 1301 suspension and its fermented milk restored the abundance and diversity of the microorganism species in the rat intestinal tract, and increased significantly. The Chaol/ACE and Shannon decree (P0.05) of the intestinal flora and the number of Simpson (P0.05) were reduced, and the reproduction of Lactobacillus in the intestinal tract of rats was promoted and the growth of Escherichia coli / Shigella was inhibited. The relative abundance of the rumen and the content of TG in the sera, the relative abundance of Dorea spp and the T in the serum in the intestine of rats. The relative abundance of C and LDL-C showed significant positive correlation with the content of HDL-C in serum (P0.05), while the relative abundance of Vibrio butyrate was significantly negatively correlated with the content of TC and LDL-C in serum (P0.05).
【學(xué)位授予單位】：揚(yáng)州大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：R151.1

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