龚树生
内耳免疫及自身免疫性内耳病研究
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- 姓名:龚树生
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学术头衔:
博士生导师
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学科领域:
耳鼻咽喉科学
- 研究兴趣:内耳免疫及自身免疫性内耳病研究
龚树生教授,男, 1962年5月生,中共党员,医学博士,主任医师,教授,博士生导师。1992年获原同济医科大学医学博士学位,2003年11月至2004年10月赴美国加州大学圣地亚哥分校做访问学者,主要从事耳鼻咽喉科临床和内耳免疫方面的研究工作。现任华中科技大学同济医学院附属协和医院耳鼻咽喉科副主任,华中科技大学同济医学院副院长,中华耳鼻喉科学会中青年委员,湖北省耳鼻喉科学会副主任委员,武汉市耳鼻咽喉科学会主任委员,《临床耳鼻咽喉科杂志》副主编,《耳鼻咽喉—头颈外科》和《听力学与言语疾病杂志》等多种学术杂志编委;高等医学院校专科教材《耳鼻咽喉科学》副主编,全国高等医药院校统编教材《耳鼻咽喉科学》编委。曾荣获1998年赛克勒中国医师年度奖优秀奖,武汉市江汉地区2001年“十大杰出青年”,2002年获教育部“高校青年教师奖”等奖项。近年来,主持承担国家自然科学基金面上项目5项、教育部高等学校优秀青年教师教学和科研奖励计划项目,以及省部级科研项目5项,在内耳免疫及自身免疫性内耳病研究方面取得了一系列研究成果,分别在Acta Otorhinolaryngol、Ann Otorhinolaryngol、Chin Med J、中华耳鼻咽喉科杂志、临床耳鼻咽喉科杂志等期刊发表学术论文60余篇,其中SCI收录5篇。龚教授在临床主攻耳科疾病的病因、发病机制、诊断和治疗,擅长耳显微外科学,尤其是在中耳疾病的外科治疗与听力重建术、面神经疾患的外科治疗、眩晕的外科治疗等领域多有建树。
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龚树生, 曾新力, 汪吉宝, 黄翔
Chin J Otorhinolaryngol, June 1998, Vol 33, No.3,-0001,():
-1年11月30日
目的 探讨细胞间粘附分子-1(intercellular adhesion molecule-1, ICAM-1)在内耳免疫反应中的作用。方法 采用钥孔 血蓝蛋白激发已全身致敏的26只大鼠内耳,诱发其内耳免疫反应,然后通过免疫组化技术检测内耳的ICAM21的表达。结果 内耳激发后6小时,螺旋轴静脉及其回流小静脉即有ICAM21表达,12小时内淋巴囊及其囊周区出现ICAM21表达,在24~48小时内耳各部位ICAM21表达达最高峰。72小时各种炎性细胞浸润到内耳的各个部位。随后ICAM21表达逐渐减弱,28天完全消失。结论 内耳免疫反应时,ICAM21在炎性细胞从循环系统进入内耳的过程中发挥着重要作用,调控ICAM21表达的细胞因子也可能还来自内淋巴囊以外的其他细胞。
迷路, 自身免疫, 胞间粘附分子-1, 内淋巴囊
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龚树生, Zhen Yan, MD, PhD; Ji-Bao Wang, PhD; Shu-Sheng Gong, PhD; Xiang Huang
Laryngoscope 113: September 2003,-0001,():
-1年11月30日
Objectives/Hypothesis: It has been recognized that immunological mechanisms could be involved in the pathogenesis of different inner ear disorders, such as progressive sensorineural hearing loss, Meniere's disease, and even sudden deafness. The endolymphatic sac acts as an immune control organ for the inner ear and has been considered as an effector site of the mucosa-associated lymphoid tissue. The purpose of the study was to determine the potentially immunological relationship between endolymphatic sac and Waldeyer ring equivalent, one of the most important affector organs in mucosa-associated lymphoid tissue. Study Design: Animal model. Methods: Thirty-six rats were employed. Two animals were killed for histological observation of Waldeyer ring equivalent, and another 34 animals were randomly divided into experimental and control groups and received bilateral intranasal immunizations with keyhole limpet hemocyanin or Freund adjuvant, respectively. The ears of immunized animals and control animals were examined for keyhole limpet hemocyanin-positive memory cells and immunoglobulin G-positive plasma cells with the technique of immunohistochemical analysis. The histopathological changes and cell proliferation in those ears were also assessed. Results: There were paired and organized lymphoid tissues in the lateral wall of the first choana in the rat. Keyhole limpet hemocyanin–positive lymphocytes appeared within the endolymphatic sac at 3 days after the first anti-genetic stimulus of the Waldeyer ring equivalent. Endolymphatic hydrops in the cochlea, elevated amounts of immunocompetent cells, and increased activity of cell proliferation within the endolymphatic sac were also considered after four intranasal injections of keyhole limpet hemocyanin. Conclusion: Presumably, endolymphatic sac is supplied with immunocompetent cells from Waldeyer ring equivalent and has an ability of coreaction with Waldeyer ring equivalent. Key Words: Endolymphatic sac, mucosa-associated lymphoid tissue, proliferation.
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【期刊论文】Discernment of the sensitized inner ear by peripheral immunocompetent lymphocytes
龚树生, GONG Shusheng, ZENG Xinli, YAN Zhen, and WANGJibao
Chinese Medical Journal 1999; 112 (8): 728-730,-0001,():
-1年11月30日
Objective Toinve stigate the role of peripheral immunocompetent lymphocytes during the immunere sponse in the innerear1 Methods Sensitized lymphocytes from peripheral blood, neck lymph nodes and spleen of three groups of donor animals, i1e1, group A immunized intradermally with keyhole limpet hemocyanine (KLH) and both inner ears challenged with KLH, group B received only KLH intradermally without inner ear challenge, and group Cused as control, were isolated and labelled with 51 Cr1 The labelled lymphocytes were injected intravenously into the recipients1 They were immunized intradermally with right inner ears challenged with KLH1 Twenty to 24hlater, the radioactivities in the temporal bones of the recipients were assayed to detect the infiltration of labelled cells1
labyrinth, autoimmunity, lymp hocytes, isotopelabeling
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龚树生, SHU-SHENG GONG. MD, PHD WUUHAN CITY. CHINA ZHEN YAN, MD, PHD
Ann Otol Rhinol Laryngol 111: 2002,-0001,():
-1年11月30日
To explore whetber The immune response of the inner ear could induce heat shock protein 70 (hsp70) in tile guinea pig cochlea, we established a model of autoimmune inner eat" disease (AIED) by systemically immunizing guinea pigs with homologous crude inner ear antigen (CIEAg).The immunized cochleas and normal control cochleas were examined for the expression of hsp70 by immunohistochemical analysis and in situ hybridization. It was found that 10 of 28 ears showed significant increases in hearing thresholds after the animals had been sensitized with CIEAg, and 67.9% of the ears from the immunized animals had modiolar vasculitis. The hsp70-1ike proteins were basically expressed in the normal cochleas, but dramatically increased expressions of hsp70 (perhaps also including hsp70-1ike molecules) and its messenger RNA were observed in the cochleas of the AIED models. The restltS suggest that the immune response of the inner ear induced the expression of hsp70 in The guinea pig cochleas.
autoimmune disease, cochlea, heat shock protein, sensorineural hearing loss.,
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【期刊论文】高血压对大鼠听功能及耳蜗乳酸脱氢酶、胆碱酯酶影响的研究
龚树生, 杨燕珍, 余青松, 于栋祯
中华老年医学杂志,2003,22(12):743~746,-0001,():
-1年11月30日
Objective To explore whether hypertension can accelerater aging process of the hearing organ, and to understand the relationship between hypertension and aging process. Methods Three-month-old and 12-month-old Wister rats (each group 20), and 3-month-old and 12-month-old SHRSP ratsf (each group 20), with normal tyrnpanic membreane, weighing 200-300g (3-month-old) and 40-600g (12-month-old) (Institute of Cardiovascular Disease, Chinese Academy of Medical Sciences), were selected as two control groups and two experimental groups respectively. The tail artery blood pressure was measured without injury. The threshold of auditory brainstem response (ABR) WAS MEASURED WITH spirit TM evoked potential meter (Nicolet Company)l. The enzyme histochemistry in the inner ear (LDH and ChE) was examined and their optical density were analyzed with HPLAS analytical system. Results It showed no difference in ABR thresholds and activites of LDH and ChE between 3-month-old Wister and SHRSP rats (P>0.05). The mean value of ABR threshold of 12-month-old Wister rats was higher than that of the two 3-month-old groups (P<0.05), LDH and ChE activity of the 12-month-old Wister rats at relevant section were significantly higher and lower respectively than those of the two 3-month-old group (p<0.05), whereas the mean value of ABR threshold of the 12-month-old SHRSP rats and the activities of LDH and ChE at relevant section were significantly higher than those of the wto 3-month-old groups (P<0.01). Conclusions It suggestes that presbycusis exists in 12-month-old Wister rats, and glycogenosis and abnormal secretory of neural transmitter was discerned after hypertension. All above factors may worsen the aging of the hearing system.
Rats,, inbred SHR, Cochlear, lactate dehydrogenase, Cholinesterases
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龚树生, SHU-SHENG GONG, DONG-ZHEN YU, JI-BAO WANG
Acta Otolaryngol 2002; 122: 5-9,-0001,():
-1年11月30日
Crude inner ear antigen (CIEAg) can induce autoimmune inner ear disease (AIED) although it is not known which subcomponent of CIEAg is involved. In this study, we investigated the relationship between 3 puried inner ear antigens (31, 42-45 and 60kD proteins) and AIED, and determined their distribution in normal guinea pig cochlea. Three groups of guinea pigs were immunized with the three inner ear antigens and one group served as a control. The hearing thresholds, serum IgG level and morphological changes in the inner ear were observed. The expression of the three antigens in the cochlea was detected using immunohistochemical techniques. No obvious changes in hearing thresholds or inner ear morphology were observed between the control and 42-45kD groups. Animals immunized with the 31 or 60kD proteins showed a signi cant increase in hearing thresholds (p B 0.05 vs control), accompanied by morphological changes in the inner ear. The serum IgG level was increased signi
autoimmune disease,, isolation and puri cation,, labyrinth.,
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龚树生, GONG Shusheng and YU Dongzhen
Chinese Medical Journal 2001; 114 (10): 1102-1104,-0001,():
-1年11月30日
Objective To isolate and purify the subcomponents of guinea piginner ear antigens for further study on the autoimmunity of the inner ear1 Methods Inner ear tissues were homogenized in pho sphate buffered saline containing 011%SDS, and then frozen and defro sted repeatedly to extract inner ear antigens1 Preparative polyacrylamidegel electrophore sis was used to separate the subcomponents of inner ear antigens1 Following electrophore sis, the protein bands were localized by rapid staining and de staining1 Res ults The major protein bands were clearly distinct when 3mg of crude inner ear antigens was loaded, and the three major subcomponents (31, 42-45 and 60 kD proteins) accounted for about 25199%, 21191% and 21110%, respectively1 Conclusion Preparative polyacrylamide gel electrophore sis can be used to purify the major subcomponents of inner ear antigens1
labyrinth, antigen, electrop horesis, polyacrylamidegel
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【期刊论文】利诺吡啶对豚鼠耳蜗外毛细胞和支持细胞钾电流的影响
龚树生, 常青, 丁娟, 唐明.
中华耳鼻咽喉科杂志,2003,38(4):263~267,-0001,():
-1年11月30日
ll recording, Ik (tetraethylammonilum—sensitive outward potassium currents) and Ikn (inward potassium currents activated at negative potential) were measured befor and fafter the administration of linopirdine in OHCs. Simultaneously, Ik (the outward rectifier potassium current)was recorded in Deiters cells. Results Ik was partly reduced after the administration of linopirdine in OHCs, moreover, Ikn was totally inhibited. However, Ik in Deiters cells was not decreased. Conclusion KCNQ channel family could only involve in construchting the variant potassium channels in outer hair cells, but not in Deiters cells.
Potaasium channels, Hair cells,, outer, Labyrinth supporting cells, Patch-clamp techniques, Diethylamiues, Linopirdine
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龚树生, 于栋祯, 杨燕珍, 黄翔, 汪吉宝
中华耳鼻咽喉科杂志,2001,36(4):285~288,-0001,():
-1年11月30日
目的探讨3种纯化的内耳抗原与自身免疫性内耳病的关系,并确定其在耳蜗的表达。方法以粗制内耳抗原的3种亚组份(31000,42000-45000和60000蛋白)作为抗原,分别免疫动物(B、C和D组),观察听阈、血清IgG水平和内耳形态学的改变,并应用免疫组织化学方法确定其在耳蜗的表达。A组为对照组,以不含抗原的聚丙烯酰胺凝胶匀浆液代替内耳抗原。结果免疫前各组听阈差异无显著性(F=0.07,P>0.05),免疫后对照组和= 组听阈和内耳形态学未见明显改变,B组(9/30只)和>组(7/28只)部分动物出现听力损失。组间听阈差异有显著性(F=9.12,P<0.01)。实验组动物血清IgG均显著性升高(F=7.46,P<0.01),B、C和D组与对照组相比差异有显著性。31000蛋白完全分布于耳蜗神经纤维,而42000-45000和60000蛋白分布广泛,螺旋神经节、Corti器、血管纹和螺旋韧带均有分布。结论粗制内耳抗原中31000和60000亚组份均能独立诱发自身免疫性内耳病,31000蛋白具有很高的组织特异性,可能作为一种标志性蛋白用于自身免疫性内耳病的临床诊断。
耳疾病, 自身免疫疾病, 诊断技术, 耳科, 自身抗原, 迷路
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龚树生, 曾新力, 汪吉宝, 黄翔
中华耳鼻咽喉科杂志 ,1998,33(1):14~16,-0001,():
-1年11月30日
目的 了解外周免疫活性淋巴细胞在内耳免疫反应中的作用。方法 提取全身致敏加双侧内耳致敏(A 组)、全身致敏(B 组)和对照组(C 组)等3组供者动物(豚鼠)的外周淋巴细胞,并在体外行51Cr核素标记。将标记的淋巴细胞分别转移到具有相同免疫史的(即全身致敏加右侧内耳致敏)受体豚鼠内,按不同的供者来源,受体动物亦分为A 组(15只)、B组(10只)、C组(10只)。20~24小时后取出受体动物双侧颞骨,称重,用C计数器测定其放射性含量(CPM/g)。结果 CPM/g平均值A、B两组致敏耳(右耳777134和698135)明显高于非致敏耳(左耳600132和553142)(P值均<0.05),C组两耳间差异无显著性(P>0.05)。A组致敏内耳与非致敏内耳CPM/g的比率为1.34, 明显高于B组(1.12)和C组(1.02),差异有显著性(P<0.05)。结论 内耳在抗原刺激下产生免疫反应受外周循环系统的免疫监控。
迷路, 自身免疫, 淋巴细胞, 同位素标记
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