肾上腺细胞培养与移植

肾上腺细胞能在体外培养,取材方便。通过体外培养人或动物的肾上腺细胞,可揭示肾上腺皮质或髓质细胞的分泌功能,进行皮质或髓质疾病的研究,并能应用于临床。培养肾上腺皮质或髓质细胞并将其移植已经成为治疗皮质激素分泌功能不全、髓质增生等疾病的重要方法。     

体外培养人胎肾上腺髓质家兔脑内移植的研究

培养妊娠16、20和24周人胎肾上腺髓质组织2周,观察组织细胞形态和功能变化。结果提示,妊娠24周人胎肾上腺髓质组织功能较妊娠16和20周者为好。培养8天的妊娠24周人胎肾上腺髓质组织移植至24只成年家兔脑内。术后1周检查4只动物,3只移植物存活良好,1只部分存活。1个月时检查10只动物,6只移植物存活良好,1只部分存活,3只移植物消失。3个月时检查10只动物,5只移植物存活良好,1只部分存活,4只移植物消失。移植物存活良好者具有儿茶酚胺(CA)合成分泌功能。移植物部分存活及消失者可能与移植过程损伤和排异反应有关。     

脑移植治愈帕金森氏病质疑

墨西哥一研究小组声称脑移植能治愈帕金森氏病。但有些科学家用此方法进行了动物试验,他们认为此方法尚不成熟。 Ignacio Madrazo等切除了因帕金森氏病引起严重丧失生活自理能力并对药物治疗无效患者的一侧肾上腺。然后将肾上腺髓质植入病人脑中。肾上腺髓质细胞与神经细胞在胚胎期有相似起源。在正常情况下,肾上腺髓质细胞产     

用微囊牛肾上腺髓质细胞移植治疗恶性肿瘤疼痛研究

用胶原酶消化法分离牛肾上腺髓质细胞,用海藻酸钠-聚赖氨酸-海藻酸钠(APA)法包埋牛肾上腺髓质细胞,将微囊牛肾上腺髓质细胞植入恶性肿瘤病人的珠网膜下腔,移植后不使用免疫抑制剂。结果显示在30例恶性肿瘤患者中,显效率为67％(20例),有效率为27％(8例),无效率为6％(2例),镇痛作用维持时间长达150天,除个别病人发烧和腰痛外未发现其它明显副作用。     

脑组织移植的现状

自1982年Backland用自体肾上腺髓质移植治疗震颤麻痹以来,我国也分别采用自体或胎儿肾上腺髓质脑内移植治疗本病。我国利用胎脑组织做供体有相对有利条件,自1986年开始,北京、上海等十二个省市用脑组织移植治疗,震颤麻痹、扭转痉挛、大脑萎缩、小脑萎缩、脑血管病后遗症、尿崩症、脊髓损伤等300多例。有的单位经动物实验研究证实移植胎鼠的脑组织在宿主鼠脑内不仅成活,而且有神经介     

人体胎脑移植治疗震颤麻痹的尝试

左旋多巴治疗震颤麻痹疗效不甚满意,为探索新的治疗途径,Backlund等近年对震颤麻痹患者进行了自体肾上腺髓质脑内移植的临床试验,手术取得一定疗效。鉴于肾上腺髓质产生多巴胺甚少,尤其是老年人细胞生物功能减退,且该手术须作肾上腺切除术,因此临床应用有一定局限。胎脑移植治疗高等动物震颤麻痹模型已获成功,然尚未见临床试验报告,我们于1985年8月进行了胎脑黑质脑内移植的临床尝试。 患者,男性,54岁,四肢震颤、强直七年,药物疗效差。面具脸,两上肢搓丸样运动,四肢肌张力增高,     

细胞凋亡调控基因bcl-2和bax在肾上腺髓质增生大鼠肾上腺组织中的表达与意义

目的研究肾上腺髓质增生大鼠肾上腺组织细胞凋亡抑制基因bcl-2和促进基因bax在肾上腺髓质增生发病机制中的作用。方法用免疫组化SP法对肾上腺髓质增生大鼠和正常对照组大鼠的肾上腺组织细胞凋亡相关基因bcl-2和bax的表达进行检测。结果bcl-2的表达,肾上腺髓质增生大鼠组阳性细胞数为(89.3±10.86)个,正常对照组为(21.2±7.65)个(P<0.05)。bax的表达,肾上腺髓质增生大鼠组阳性细胞数为(16.1±4.62)个,正常对照组为(54.8±9.31)个(P<0.05)。结论细胞凋亡调控基因参与了肾上腺髓质增生的病理过程。在此调控网络中,bcl-2基因发挥了较为重要的作用,bax基因的表达减少。     

体外培养胎儿肾上腺髓质脑内移植治疗帕金森病的临床研究

采用脑立体定向技术,移植培养的人胎肾上腺髓质组织至17例重症帕金森病患者脑内尾状核头部。移植术前后进行Webster 症状评分,头颅CT 检查,腰穿脑脊液DA 测定,血免疫球的蛋白、补体和T 淋巴细胞及其亚群检查,EEG、SEP 和VEP 检查,神经心理学测验和服药方面观察,随诊3～18个月。结果提示胎儿肾上腺髓质组织在脑内存活良好,临床效果比较满意,比目前其他移植治疗方法具有临床可行性和可靠性。     

第五讲 肾上腺（一）

第一节肾上腺髓质一、肾上腺髓质的解剖、组织学特征肾上腺位于脊椎两旁,两肾上端。肾上腺是由两种不同组织组成的。外层为皮质,内层为髓质。从胚胎发生看,皮质来自中胚层,而髓质的嗜铬细胞则来自外胚层。如果固定液中含有铬盐,髓质细胞即呈棕黄色颗粒,故称为嗜铬细胞。肾上腺皮质和髓质,在人体内虽然合为一个腺体,但胚胎来源     

同种异体大白鼠肾上腺髓质移植治疗震颤麻痹实验性研究

本研究系将异体大白鼠的肾上腺髓质细胞悬液注入用6—OH—DA 损伤一侧黑质而造成震颤麻痹大白鼠动物模型的尾状核内。随着存活时间的不同,观察震颤麻痹大白鼠的旋转行为逐渐改善,以至最后旋转行为停止。同时用石腊切片,光学显微镜观察。受体内被移入的异体大白鼠肾上腺髓质细胞已经成活,并成团簇状。     

Transplant of cultured neuron-like differentiated chromaffin cells in a Parkinson's disease patient. A preliminary report

BACKGROUND: Treatment of Parkinson's Disease (PD) has been attempted by others by transplanting either the patient's own adrenal medullary tissue or fetal substantia nigra into caudate or putamen areas. However, the difficulties inherent in using the patient's own adrenal gland, or the difficulty in obtaining human fetal tissue, has generated the need to find alternative methods. METHODS: We report here of an alternative to both procedures by using as transplant material cultured human adrenal chromaffin cells differentiated into neuron-like cells by extremely low frequency magnetic fields (ELF MF). RESULTS: The results of this study show that human differentiated chromaffin cells can be grafted into the caudate nucleus of a PD patient, generating substantial clinical improvement, as measured by the Unified Rating Scale for PD, which correlated with glucose metabolism and D2 DA receptor increases as seen in a PET scan, while allowing a 70% decrease in L-Dopa medication. DISCUSSION: This is the first preliminary report showing that transplants of cultured differentiated neuron-like cells can be successfully used to treat a PD patient.     

警惕脑内异体组织移植产生免疫排斥反应

采用NIH标准补体依毫蔓的细胞毒法检测7例接受胚胎肾上腺髓质组织脑内移植患者血中抗T、B淋巴上的HLA-A、B、C、DR抗体。每1例患者血清对16名正常人T、B淋巴细胞进行微量细胞毒实验,结果1例患者血清呈HLA-DR抗体阳性反应。结果提示在开展胚胎组织脑内移植工作中应加强对免疫排斥反应的监视。     

自体肾上腺髓质移植治疗震颤麻痹的实验性研究

本文选经6—OHDA破坏一侧大白鼠的黑质,制成震颤麻痹的动物模型16只,择取其左侧肾上腺,制成肾上腺髓质细胞悬液,分2～4点注入自体损伤侧尾状核头部。术后成活1～3个月,有11只实验动物行为明显改善,移植尾状核内的肾上腺髓质细胞已成活,在注射部位聚集成若干细胞(?)簇。     

Adrenal Cortical and Medullar Hyperplasia—A Retrospective Analysis of 6 Cases

The features of the symptoms,laboratory tests and pathological characteristics of adrenal cortical and medullary hyperplasia were studied.In 6 cases of hypercatecholaminenia,plasma norepinephrine(NE),epinephrine(E),catecholamine(CA) and 24-h urinary vanillylmandelic acid(VMA),17-hydroxycorticosteroid (OHCS) and 17-ketosteroid (KS) were determined.adrenal glands were examined by CT scan and ^131I-MIBG imaging.Pathological examination was performed after operation.The results showed that in 6 cases of hypercatecholaminenia (3 men and 3 women) aged from 34-50 years,the clinical features were just like “pheochromocytoma”,for example,episodic headache,perspiration,palpitation,pallor,apprehension,nausea,tremor,anxiety and so on.Plasma levels of CA,NE and E were elevated in all 6 cases.24-h urinary samples obtained at the onset revealed elevated VMA in 1 case.24-h urinary cortisol was obviously elelated in all 6 cases.24-h urinary 17-OHCS,17-KS was normal.B-type ultrasound,CT,MRI and ^131I-MIBG revealed 9 lateral adrenal gland diffuse or nodular enlargement in 6 cases.Pathologic examination showed adrenal cortical and medullary hyperplasia.Clinically,adrenal cortical and medullary hyperplasia resembled “pheochromocytoma”.The most significant feature of this disease was both elevated plasma CA and 24-h urinary cortisol obviously.Pathologic examination showed adrenal cortex nodular hyperplasia and medullar diffuse or limit hyperplasia.Whether it is an independent disease or symptoms of the other disease has not final conclusion up till now.     

Multiple endocrine neoplasia type 2b: long-term follow-up of a case

Multiple endocrine neoplasia type 2b is a rare inherited syndrome which comprises the association of medullary thyroid carcinoma, phaeochromocytoma, widespread neuromatous proliferation and a characteristic body habitus. In this report we present the late clinical course and autopsy findings of the first patient with this syndrome described in Australia. At presentation she was found to have a right adrenal phaeochromocytoma and medullary thyroid carcinoma which were resected in separate operations. No clinical or biochemical evidence of residual medullary thyroid carcinoma was identified in life. However, in spite of serial vanillylmandelic acid estimations, which showed normal or only mildly elevated levels, and normal results of urinary catecholamine studies, a left adrenal phaeochromocytoma was identified in a metaiodobenzylguanidine (MIBG) study performed 14 years after presentation. Her late clinical course was dominated by progressive dysphagia, intestinal dysmotility and megacolon associated with unrelenting malnutrition. After her death due to an intracerebral haemorrhage, an autopsy confirmed the presence of a left adrenal phaeochromocytoma and revealed diffuse intestinal ganglioneuromatosis to be the cause of her intestinal dysmotility. No residual medullary thyroid carcinoma was found. This case emphasises the propensity for multiple endocrine tumours in these patients and highlights the potentially significant role of intestinal ganglioneuromatosis in the natural history of this condition.     

Ultrastructural and biochemical characterization of catecholamine release mechanisms in cultured human pheochromocytoma cells

OBJECTIVE: To characterize ultrastructurally and biochemically catecholamine release mechanisms of cultured human pheochromocytoma cells in the basal and stimulated states. METHODS: The cultured pheochromocytoma cells were prepared from human adrenal pheochromocytoma tumors. Biochemical determinations of catecholamine secretion from the cultured cells were carried out in the basal and stimulated states. Transmission electron microscopy was used to observe the modes of catecholamine release from the cells without and with stimulation by depolarization of the cells with the administration of 50 mmol/L KCl. RESULTS: Biochemical determinations consistently showed spontaneous secretion of catecholamines from the cultured cells in the basal state without stimulation. Catecholamine release in a calcium-dependent manner could be enhanced in the cells in response to high extracellular potassium concentration. A series of electron microscopic observations of the cultured cells consistently disclosed the classical exocytotic profiles on the cell surface in the basal state. In addition to abundant increase in the number of classical single exocytosis, compound exocytosis was frequently observed in the stimulated cells. Furthermore, other modes of catecholamine release mechanism involving the formation of pseudopodial and/or tubule-like structures, which were different from classical exocytosis, were often present in the intensely stimulation cells. CONCLUSIONS: Based on the biochemical and electron microscopic findings, we concluded: (1) classical single exocytosis is considered to be a primary mechanism responsible for spontaneous secretion of catecholamines from the cells in the basal state; (2) compound exocytosis is an essential mechanism for extruding large amounts of catecholamines in the stimulated cells; and (3) other modes of catecholamine release mechanism may operate in the cells in response to intense stimulation. These morphological data may be helpful in explanation of biochemical variability and extreme diversity of clinical manifestations in patients with pheochromocytoma tumor.     

Adrenal cortical and medullar hyperplasia—A retrospective analysis of 6 cases

Summary The features of the symptoms, laboratory tests and pathological characteristics of adrenal cortical and medullary hyperplasia were studied. In 6 cases of hypercatecholaminenia, plasma norepinephrine (NE), epinephrine (E), catecholamine (CA) and 24-h urinary vanillylmandelic acid (VMA), 17-hydroxycorticosteroid (OHCS) and 17-ketosteroid (KS) were determined. Adrenal glands were examined by CT scan and¹³¹I-MIBG imaging. Pathological examination was performed after operation. The results showed that in 6 cases of hypercatecholaminenia (3 men and 3 women) aged from 34–50 years, the clinical features were just like “pheochromocytoma”, for example, episodic headache, perspiration, palpitation, pallor, apprehension, nausea, tremor, anxiety and so on. Plasma levels of CA, NE and E were elevated in all 6 cases. 24-h urinary samples obtained at the onset revealed elevated VMA in 1 case. 24-h urinary cortisol was obviously elevated in all 6 cases. 24-h urinary 17-OHCS, 17-KS was normal. B-type ultrasound, CT, MRI and¹³¹I-MIBG revealed 9 lateral adrenal gland diffuse or nodular enlargement in 6 cases. Pathologic examination showed adrenal cortical and medullary hyperplasia. Clinically, adrenal cortical and medullary hyperplasia resembled “pheochromocytoma”. The most significant feature of this disease was both elevated plasma CA and 24-h urinary cortisol obviously. Pathologic examination showed adrenal cortex nodular hyperplasia and medullar diffuse or limit hyperplasia. Whether it is an independent disease or symptoms of the other disease has not final conclusion up till now     

人胎儿角膜缘干细胞的体外培养及鉴定

目的 探讨人胎儿角膜缘干细胞的体外培养及鉴定方法。方法 应用消化培养法对人胎儿角膜缘上皮组织进行体外培养并观察记录培养细胞的生长特性；对胎儿角膜缘组织及培养细胞采用一系列单克隆抗体进行免疫酶细胞化学染色检测；并用扫描电镜观察原代细胞的表面状况。结果 胎儿角膜缘上皮细胞原代培养时生长旺盛，传代培养时保持高增殖速率。免疫酶细胞化学染色见培养前胎儿角膜缘组织基底层有多量AE5阴性、AE1和PCNA阳性细胞，并见较多HLA-DR阳性细胞；原代细胞绝大部分AE5和HLA-DR染色阴性，AEI和PCNA阳性；传代培养至第3代时AE5阴性细胞仍占大部分；扫描电镜观察原代细胞以球形或短圆柱形为主，表面多突起和微绒毛。结论 采用消化培养法可成功培养出具有高增殖力和低抗原性的人胎儿角膜缘干细胞．为临床眼表重建开辟了新的思路。     

Thyroid and adrenal relationships

A brief review of the actions of adrenal medullary and thyroid hormones is presented and the ways in which they interact are examined. It is concluded that thyroid hormone produces the necessary intracellular environment without which the steady state and emergency actions of cathecholamines would be vitiated. In hyperthyroidism the increased concentration of thyroid hormones results in a lowering of the threshold for catecholamine action. For this reason it is possible to alleviate many of the symptoms of thyrotoxicosis by means of drugs which block β-adrenergic receptors.     

人胚胎肾间质成纤维细胞培养及其鉴定

目的：探讨人胚胎肾脏成纤维细胞培养方法。方法：利用人胚胎肾脏肾髓质进行培养，然后通过细胞形态学，超微结构，免疫细胞化学等方法进行分析鉴定。结果：培养的细胞为长梭形，单核，超微结构为典型的成纤维细胞，并表达成纤维细胞表面抗原，波形蛋白，结蛋白，不表达角蛋白。结论：成功地培养出人胚胎肾间质成纤维细胞。