微囊化异种甲状旁腺组织移植的研究

目的探讨微囊化异种甲状旁腺(PTG)组织移植对Wistar大鼠甲状旁腺功能低下的治疗作用及微囊的通透性。方法 40只去甲状旁腺Wistar大鼠随机分为微囊组、非微囊组、空囊组、空白对照组。用海藻酸-钡交联微囊包裹兔甲状旁腺组织,移植至Wistar大鼠肾包囊。移植后每隔2周取血测血钙,移植后第16周取移植物进行透射电镜检查及T淋巴细胞、大鼠IgG抗体的免疫组织化学染色。结果微囊组移植后第4周血清钙由(1．62±0．04)mmol／L恢复至正常水平 (2．2～2．6)mmol／L,9例维持至观察期结束(P<0．01),非微囊组、空囊组及空白对照组血清钙差异无统计学意义(P>0．05)。第16周取出移植物检测显示移植物活性良好,微囊周围可见较多T 淋巴细胞浸润,囊壁及囊内IgG抗体染色阳性。结论海藻酸-钡交联微囊可对甲状旁腺组织起到有效的保护作用,使甲状旁腺组织较长时间存活并发挥正常功能。但海藻酸-钡交联微囊并未减轻受体免疫排斥反应的激活且未有效的免疫隔离IgG抗体。     

甲状旁腺瘤组织同种移植治疗甲状旁腺功能低下症初步报告

目的　为甲状旁腺移植的供体来源提供一种新途径。　方法　将甲状旁腺瘤组织用 6Gy60 钴照射后 ,切成 0 5mm× 0 5mm× 0 5mm～ 1 0mm× 1 0mm× 1 0mm薄片 ,植入裸鼠肾包膜下过渡 14d ,移植给甲状旁腺功能低下症患者 ,受者术前 1d及术后 14d服用环孢素A(5mg·kg 1·d 1) ,以后不用任何免疫抑制剂。术后观察临床症状、体征、治疗的钙剂及罗钙全剂量变化 ,检测血钙及血甲状旁腺激素 ,以确定移植效果。　结果　临床应用 5例 ,其中 3例术后不需静脉补钙 ,口服钙剂量及罗钙全剂量减少 ,血钙浓度基本恢复到正常值。　结论　甲状旁腺瘤组织为甲状旁腺移植提供了一种方便、易于获取的新的供体 ,且可移植给多个患者治疗。     

微囊化新生猪甲状旁腺细胞异种移植的实验研究

目的 探讨微囊化新生猪甲状旁腺细胞异种移植治疗大鼠甲状旁腺功能低下症的效果。方法 应用微囊化技术，制备微囊化（海藻酸钠－聚赖氨酸－海藻酸钠生物微胶囊）新生猪甲状旁腺细胞，32只去甲状旁腺的Wistar大鼠随机分成微囊组、非微囊组、空囊组和对照组，分别移植微囊化新生猪甲状旁腺细胞、甲状旁腺细胞、空微囊及生理盐水。移植后监测血钙及甲状旁腺素水平40周，40周后回收移植物，透射电镜检查。结果 移植后，微囊组大鼠血钙及甲状旁腺素水平恢复到正常范围内，直至观察结束时（40周），透射电镜检查显示移植物存活良好；非微囊组、空囊组和对照组大鼠的血钙及甲状旁腺素水平无改善。结论 微囊化新生猪甲状旁腺细胞异种移植在不用免疫抑制剂情况下，可以在大鼠体内存活，且有功能；海藻酸钠－聚赖氨酸－海藻酸钠生物微胶囊对免疫活性细胞及抗体具有屏蔽作用。     

同种异体甲状旁腺组织移植治疗甲状旁腺功能低下的实验研究

目的 探讨甲状旁腺经不同预处理并移植于腹直肌内后甲状旁腺移植物的功能和生存情况.方法 成年雄性Wistar大鼠70只作为供体,成年雄性SD大鼠35只作为受体,建立去甲状旁腺模型受体后采用随机数字表法随机分为直接移植组、高氧培养移植组、环孢素A组、60Co照射移植组及综合处理组5组,每只受体接受2只供体的4个甲状旁腺组织块,甲状旁腺移植于大鼠的腹直肌内.观察各组大鼠在甲状旁腺移植前后不同时相血清钙和甲状旁腺激素(PTH)的变化.结果 各组在甲状旁腺组织移植后,1周内血清钙和PTH均能达到或接近正常水平,与移植前相比差异均有统计学意义(P＜0.01).各组移植物的存活时间不同,直接移植组的存活时间最短,血清钙和PTH维持正常水平的时间分别为3周和4周.高氧培养移植组、环孢素A组、60Co照射移植组以及综合处理组血清钙及PTH维持在正常或接近正常水平的时间分别为5周和8周、6周和8周、5周和7周以及5周和9周;除高氧培养移植组移植术后9周(血清钙)及60Co照射移植组移植术后8周(PTH)差异无统计学意义外,其余4组的血清钙和PTH水平在移植术后4～9周与直接移植组比较其差异均具有统计学意义(P＜0.05),直接移植组较低;高氧培养移植组、CsA组和60Co照射移植组的血清钙和PTH水平在移植术后7～9周低于综合处理组(P＜0.05),综合处理组的血清钙和PTH的维持时间较长.结论 腹直肌内甲状旁腺移植能维持血钙于正常水平;甲状旁腺移植物或受体经预处理后能延长其存活时间.甲状旁腺组织经培养后再移植是治疗甲状旁腺功能减退症的一条有效途径.     

人胚甲状旁腺细胞移植治疗甲状旁腺功能低下症

目的　探讨人胚甲状旁腺细胞移植治疗甲状旁腺功能减退症的临床意义。　方法　将培养的人胚甲状旁腺细胞在B超引导下移植到 6例甲状旁腺功能减退症患者的肾周围脂脂囊中。应用放射免疫方法测定患者术前、术后血中甲状旁腺激素 (PTH)及钙水平 ,并进行自身对照 ,对疗效进行评价。　结果　 6例患者在接受人胚甲状旁腺细胞移植前 ,血钙及PTH平均水平分别为 (1 6 3±0 12 )mmol/L及 (1 36± 0 2 1)ng/L ;接受移植 3d后分别为 (1 77± 0 2 2 )mmol/L及 (9 5 3± 2 2 1)ng/L ,两者差异有显著性意义 (P <0 0 1) ;6d后达正常水平 ,临床症状逐渐减轻至消失 ;术后观察 9～ 12个月病情保持稳定。　结论　培养的人胚甲状旁腺细胞肾周围脂肪囊内移植是治疗甲状旁腺功能减退症的一种较理想的新方法     

甲状旁腺激素基因治疗甲状旁腺功能低下症

目的观察甲状旁腺激素(PTH)基因和蛋白体外表达情况,并评价其基因治疗甲状旁腺功能低下模型鼠的作用。方法(1)以脂质体将质粒pcDPG分别1次和多次转染293细胞,观察绿色荧光蛋白(GFP)的表达并计算转染率;(2)转染24、48、72和96h后real-ti me定量PCR和Western blot法检测PTH基因与蛋白表达,并活性鉴定;(3)建立甲状旁腺功能低下症模型,将pcDPG质粒多次肌肉注射治疗,监测血钙和PTH值、存活时间及各器官病理变化。结果转染后24h即见GFP表达,72h达高峰,96h开始减少;多次转染后GFP表达率可达90%以上;PTHcD-NA拷贝数转染24h为5×103,72h达最高为8×104,多次转染显著增高(P<0.01);Western blot见48h和72h有PTH蛋白表达,其可对抗甲状旁腺切除小鼠抽搐症状;术后第2天血钙与PTH明显低于术前(P<0.05),pcDPG质粒大、中剂量组连续治疗48h后血钙与PTH值均恢复正常。结论重组PTH基因治疗甲状旁腺功能低下模型鼠有较好的疗效。     

造血干细胞基因治疗甲状旁腺功能低下症的实验研究

目的　探讨造血干细胞基因治疗甲状旁腺功能低下症 (HPT)的实验效果。方法　构建重组甲状旁腺素 (PTH)基因的小鼠干细胞病毒 (MSCV)重组质粒 ,转染PA31 7包装细胞 ,G41 8筛选阳性克隆 ,获得的重组病毒液感染造血干细胞 ,静脉注入HPT小鼠血中 ,检测各组小鼠症状改善情况、血PTH及血钙变化情况。结果　获得滴度为 2× 1 0 7CFU(集落形成单位 ) /ml的分泌人PTH的浓缩病毒悬液 ,1× 1 0 6 个细胞培养 48h时PTH的分泌量为 1 5ng。经聚合酶链反应 (PCR)扩增未检测到有野生型病毒存在 ,可以安全应用。感染造血干细胞输注后 ,实验组动物未再出现HPT临床表现 ,而且血钙及血PTH均可长期保持在接近正常值范围内 ,较单纯注射浓缩病毒悬液具有更好的疗效。结论　获得MSCV PTH重组质粒及高滴度的分泌人PTH的浓缩病毒悬液。整合有PTH基因的造血干细胞输注后达到较长期治愈小鼠HPT ,为进一步HPT的临床基因治疗提供了可靠依据。     

小鼠胚胎干细胞转基因治疗甲状旁腺功能低下症的实验研究

目的　探讨小鼠胚胎干细胞 (TC 1)转基因治疗甲状旁腺功能低下症 (HPT)。方法包装出重组人甲状旁腺素 (PTH )基因的小鼠干细胞病毒 (MSCV) ,以其感染小鼠ESCs ,检测基因转导效率 ,PTH分泌情况 ;观察体内外分化情况 ,以及注入模型鼠体内各组小鼠血PTH和血钙变化情况。结果　获得滴度为 2× 10 7集落形成单位 (CFU ) /ml的重组逆转录病毒 ,经聚合酶链反应(PCR)扩增未检测到有野生型病毒存在 ,可以安全应用。感染TC 1的效率为 70 % ,每 10 6未分化TC 1每 48h分泌PTH约 10ng。重组有PTH基因的TC 1在体内外均可分化出三胚层组织 ,注入模型鼠体内 ,在观察期间实验组动物未再出现甲旁低表现 ,而且血PTH和血钙均保持在接近正常值范围内。结论　MSCV介导外源PTH基因可高效转导TC 1并持续分泌PTH ;体内外分化实验证明TC 1具有全能性 ,而且内环境并不是决定TC 1分化的唯一因素。经基因转导的TC 1可较好的改善模型鼠的症状 ,是未来细胞移植的一种潜在来源。     

胚胎干细胞转基因治疗甲状旁腺功能低下症

目的　探讨小鼠胚胎干细胞 (TC 1)转基因治疗甲状旁腺功能低下症 (HPT)。方法包装出重组人甲状旁腺素 (PTH )基因的小鼠干细胞病毒 (MSCV) ,以lml重组病毒液加入 poly brene(终浓度 8mg/L)感染TC 1细胞 ,检测基因转导效率 ,PTH分泌情况 ;以及每 1× 10 5个 /mlTC 1细胞注入模型鼠体内各组小鼠血PTH和血钙变化情况。结果　获得滴度为 2× 10 7集落形成单位 (CFU) /ml的重组逆转录病毒 ,其感染TC 1的效率为 70 % ,每 10 6 未分化TC 1每 48h分泌PTH10ng。重组有PTH基因的TC 1细胞注入模型鼠体内后 ,在观察期间实验组动物血PTH和血钙均保持在接近正常值范围内。结论　MSCV介导外源PTH基因可高效转导TC 1并持续分泌PTH ;内环境并不是决定TC 1分化的唯一因素。经基因转导的TC 1可较好的改善模型鼠的症状 ,是未来细胞移植的一种潜在来源。     

甲状旁腺功能低下患者细胞移植及其护理

目的探讨人胚甲状旁腺细胞移植治疗甲状旁腺功能减退症的临床意义。方法将培养的人胚甲状旁腺细胞在B超引导下移植到12例甲状旁腺功能减退症患者的肾周围脂肪囊中,采用放射免疫方法测定患者术前、术后血中甲状旁腺激素及钙的水平,并进行自身对照,对疗效进行评价。结果12例患者在接受人胚甲状旁腺细胞移植后,临床症状逐渐消失。结论培养的人胚甲状旁腺细胞肾周围脂肪囊内移植是治疗甲状旁腺功能低下的一种理想的新方法,而科学的护理是移植成功的重要保证。     

Xenotransplantation of Microencapsulated Canine Islets into Diabetic Rats

Abstract: Islets of Langerhans were isolated in high yields from canine pancreata. In the procedure, the pan-creata were perfused and digested with collagenase, and the islets were then purified on histopaque density gradients. As many as 60,000 islets were isolated from a single pancreas. Islets were encapsulated in alginate-polylysinealginate membranes with the aid of an air-jet droplet generator. In vitro studies demonstrated that the isolated and encapsulated islets secreted insulin in response to glucose and IBMX challenge for at least 9 weeks. In in vivo studies 6 diabetic Wistar rats were transplanted with 5,000 to 8,000 encapsulated islets each. The diabetic condition was reversed in all recipients for up to 112 days. In control animals, which received free, unencapsulated islets, the xenografts remained functional for fewer than 21 days. Microcapsules retrieved from normoglycemic transplant recipients 1 and 2 months posttransplantation were shown to contain viable islet tissue, and no cellular overgrowth was observed on capsular surfaces. The results of the study indicate a considerable clinical potential of microencapsulated canine islet xenografts.     

大鼠甲状旁腺细胞经培养后再移植的实验研究

目的　研究大鼠甲状旁腺细胞经培养后再移植对其存活的影响。方法　将经胶原酶和胰蛋白酶消化的甲状旁腺细胞培养后再行移植 ,观察移植物的存活情况 ,并对移植物做透射电镜观察。结果　新鲜甲状旁腺移植组平均存活期为 (9.2 5± 3.4 5 )d ;甲状旁腺细胞培养后移植 ,移植物的存活时间为 (46 .2 5± 7.4 4 )d ,明显延长(P<0 .0 1) ,在 5 0d观察期内 ,8只鼠中有 6只的血清钙及PTH值持续在正常范围内。移植物内可见完整的甲状旁腺细胞 ,其内见丰富的粗面内质网、杆状的线粒体及分泌颗粒。结论　大鼠甲状旁腺细胞经培养后再移植可以延长移植物的存活时间 ,是治疗甲状旁腺功能低下症的一条有效途径。     

Allotransplant of Microencapsulated Parathyroid Tissue in Severe Postsurgical Hypoparathyroidism: A Case Report

The last therapeutic alternative in severe postsurgical hypoparathyroidism is allotransplantation of microencapsulated parathyroid cells. With this technique, it is possible to implant cells or tissue of parathyroid origin to replace them in such patients, without immusupression. We report an allotransplant of parathyroid tissue in a patient with continous endovenous requirement of calcium to survive. The microencapsulation was carried out with a commercial sodium alginate. We implant 23 microspheres in the nondominant forearm and 40 microspheres in the leg in a second attempt. In this article, we show functionality of the graft for at least 20 months without requirement of endovenous calcium. We report this procedure as a therapeutical alternative in severe hypoparathyroidism.     

同种异体甲状旁腺移植治疗甲状旁腺机能减退性心肌病三例报告

利用经培养的胎儿甲状旁腺（ＰＴＧ），对３例甲状旁腺机能减退性心肌病（ＨＯＰＴＣＰ）行ＰＴＧ移植治疗。供体为４～７个月水囊引产的死胎，在显微镜下将胎儿ＰＴＧ取出，４℃Ｈａｎｋ氏液漂洗，经培养５～８天后，在Ｂ型超声波引导下，一次性将６～１０个胎儿ＰＴＧ注入受者１侧肾包膜内。术后患者症状逐渐改善，随访１年半均未复发，该方法简便易行，符合生理需要。     

腔镜下甲状旁腺全切联合部分甲状旁腺组织前臂移植治疗继发性甲状旁腺亢进

目的探讨腔镜下甲状旁腺全切（endoscopic total parathyromectomy,ETP）联合部分甲状旁腺组织前臂移植（parathyroid tissue autotranspIantafion,PTA）治疗继发性甲状旁腺亢进（secondary hyperparathyroidism,SHPT）的安全性、可行性。方法2004年6月-2009年6月72例SHPT经胸前路径行腔镜下全部甲状旁腺切除,同时将20枚1-1．5mm。大小的健康甲状旁腺组织移植于患者前臂肌膜下。结果71例ETP联合PTA顺利完成,无术中及围手术期死亡。手术时间平均119．9rain（85-155rain）,术中出血量平均39．7ml（10-60m1）,下床时间平均1．2d（0．5-2．0d）,住院时间平均4．7d（3-8d）。1例因术中出血中转开放手术,术后发现声音嘶哑,经保守治疗后缓解。术后临床症状明显改善,生化检查（甲状旁腺激素、碱性磷酸酶、血钙、血磷等指标）恢复正常或明显改善。72例随访10个月-5年,平均3．8年,2例术后甲状旁腺亢进症状复发,二次手术后痊愈。结论ETP联合PTA治疗SHPT安全、可行,疗效满意。     

Daily Treatment of Aged Ovariectomized Rats with Human Parathyroid Hormone (1-84) for 12 Months Reverses Bone Loss and Enhances Trabecular and Cortical Bone Strength

Most studies that have investigated the anabolic effects of parathyroid hormone (1-84) (PTH) or PTH fragments on the skeleton of ovariectomized (OVX) rats have evaluated the short-term effects of high-dose PTH(1-34) in young animals. This study used densitometry, histomorphometry, and biomechanical testing to evaluate the effects of 12-month daily treatment with low-dose PTH (15 or 30 μg/kg) in rats that were 10 months old at baseline, 4 months after OVX. Bone mineral density (BMD) and bone strength were reduced substantially in control OVX rats. The 15 μg/kg dose of PTH restored BMD to levels similar to those in sham animals within 6 months at the lumbar spine, distal and central femur, and whole body and maintained the BMD gain from 6 to 12 months. The 30 μg/kg dose produced greater effects. Both PTH doses normalized the trabecular bone volume-to-total volume ratio (BV/TV) at lumbar vertebra 3 but not at the proximal tibia (where baseline BV/TV was very low), solely by increasing trabecular thickness. PTH dose-dependently increased bone formation by increasing the mineralizing surface, but only the 30 μg/kg dose increased resorption. PTH increased cortical BMD, area, and thickness, primarily by increasing endocortical bone formation, and restored all measures of bone strength to levels similar to those in sham animals at all skeletal sites. PTH increased bone mass safely; there was no osteoid accumulation, mineralization defect, or marrow fibrosis and there were no abnormal cells. Thus, long-term PTH therapy normalized bone strength in the aged OVX rat, a model of postmenopausal osteoporosis, through increased bone turnover and enhanced formation of both trabecular and cortical bone.     

高能震波治疗骨与软组织疾患的初步探讨

目的：研究高能震波治疗骨与软组织疾患的机制及其临床应用。方法：我们首先用离体猪骨作为实验材料，应用ＥＳＷＬ型碎石机进行实验，并合作开发研制出骨伤治疗机，应用于临床。结果：找出高能震波的安全工作电压和冲击次数，应用骨伤治疗机治疗数百例骨科常见病，结果非常满意。结论：探讨了高能震波的基本特性和治疗机理，以及临床应用的经验     

In Vitro Hydrodynamic Evaluation of a Scaffold for Heart Valve Tissue Engineering

Although prosthetic heart valves have saved many lives, the search for a living substitute continues with the aid of tissue engineering. Much progress has been made so far, but the translation of this technology to clinical reality remains a challenge, especially due to the structural complexity of heart valves and the harsh environment they are in. In a joint effort, researchers from Federal University of ABC and Institute Dante Pazzanese of Cardiology have conceived a new bioresorbable scaffold for heart valve tissue engineering (HVTE), whose hydrodynamic performance was first assessed and described in this work. The scaffold was studied at the mitral position of a left heart simulator from Escola Politécnica of the University of São Paulo, under 60 bpm and with no cell seeding. In this condition, two‐dimensional particle image velocimetry was performed to investigate the flow during diastolic and systolic phases. The results indicate that the scaffold can withstand the required intraventricular pressures for a simulated normal physiologic condition in a bioreactor. Furthermore, the averaged (N = 150) velocity vector maps showed a smooth and well‐distributed flow during diastole and qualitatively demonstrated no‐significant regurgitation at systole.     

Effect of Long Term Treatment with Gonadotrophic Hormones on In Vitro Metabolism of [3H]Progesterone in Human Testicular Tissue

Gonadotrophic hormones appear to influence the preferred pathways for intratesticular androgen biosynthesis. Individuals with low levels of gonadotrophin in the peripheral circulation, as e. g. prepubertal boys and a hypophysectomized adult male, have previously been shown to metabolize [³H]progesterone in vitro mainly to 20α-dihydroprogesterone (20α-DH-P) and less to 17α-hydroxyprogesterone (17α-OH-P), i. e. an "immature metabolic pattern" (Berg et al. 1976; Kjessler & Berg 1976b).
With physiologically increasing amounts of circulating gonadotrophins the preferred metabolic pathway in vitro shifts in favour of 17α-OH-P, i. e. a "mature metabolic pattern". Such an alteration in the preferred in vitro metabolic pattern of [³H]progesterone has also been obtained by exogenous administration of gonadotrophins to a 47, XYY-male with an initially immature metabolic pattern (Berg & Kjessler 1976).
We have sequentially analysed the metabolism of [³H]progesterone in vitro in testicular incubates derived from a chromosomally normal male before and after 16 weeks of substitution therapy with gonadotrophic hormones.
The patient originally displayed an "immature" pattern of progesterone metabolites, i. e. the ratio 20α-DH-P/17é-OH-P was 3.04. After treatment with human menopausal gonadotrophins and hCG, the metabolic activity in total had increased, and the ratio 20é-DH-P/17é-OH-P had switched to 0.39.
These results confirm and extend the concept that gonadotrophic hormones may have a regulatory function on androgen biosynthesis by stimulating the oxidative metabolic pathway from progesterone to biologically active androgens via 17α-hydroxyprogesterone.     

Effects of Treatment of Ovariectomized Adult Rhesus Monkeys with Parathyroid Hormone 1-84 for 16 Months on Trabecular and Cortical Bone Structure and Biomechanical Properties of the Proximal Femur

Treatment of monkeys and humans with parathyroid hormone (PTH) 1-84 stimulates skeletal remodeling, which increases trabecular (Tb) bone mineral density (BMD) but decreases cortical (Ct) BMD at locations where these bone types predominate. We report the effects of daily PTH treatment (5, 10, or 25 μg/kg) of ovariectomized (OVX) rhesus monkeys for 16 months on bone structure and biomechanical properties at the proximal femur, a mixed trabecular and cortical bone site. PTH reversed the OVX-induced decrease in BMD measured by dual-energy X-ray absorptiometry at the proximal femur, femoral neck, and distal femur. Peripheral quantitative computed tomography confirmed a significant decrease in Ct.BMD and an increase in Tb.BMD at the total proximal femur and at the proximal and distal femoral metaphyses. The decrease in Ct.BMD resulted primarily from increased area because cortical bone mineral content was unaffected by PTH. Histomorphometry revealed that PTH significantly increased the trabecular bone formation rate (BFR) as well as trabecular bone volume and number. PTH did not affect periosteal or haversian BFR at the femoral neck, but cortical porosity was increased slightly. PTH had no effects on stiffness or peak load measured using a shear test, whereas work-to-failure, the energy required to fracture, was increased significantly. Thus, PTH treatment induced changes in trabecular and cortical bone at the proximal femur that were similar to those occurring at sites where these bone types predominate. Together, the changes had no effect on stiffness or peak load but increased the energy required to break the proximal femur, thereby making it more resistant to fracture.