小鼠甲状腺细胞的原代培养及其摄碘功能研究

目的 建立小鼠甲状腺细胞的原代培养方法,并研究其体外摄碘功能,为甲状腺功能的体外研究奠定基础.方法 取小鼠甲状腺组织,去筋膜和结缔组织,剪碎后用1 mg/mL的胶原酶于37℃消化2h,其间问歇振荡数次,1,200g离心收集细胞沉淀,Ficoll分离得甲状腺单个核细胞,灭菌PBS洗涤细胞2遍,用RPMI1640完全培养基调整细胞浓度至2×10 5/mL,24孔板常规接利,培养,以"半量换液法"连续培养数天.用抗甲状腺球蛋白抗体为识别抗体的间接免疫荧光法检测原代培养细胞甲状腺球蛋白的表达状况.体外摄碘试验于每孔加入0.1μci125I后继续培养2h,用冰冷的PBS洗涤细胞数遍,裂解细胞并进行蛋白浓度测定及放射性计数.温度及竞争物阻断试验分别在0℃～4℃和加入100mmol/L NaClO4的条件下进行.结果 采用上述方法获得的小鼠甲状腺原代培养细胞在体外可形成甲状腺细胞所特有的"次级滤泡样"结构.免疫荧光检测结果显示,细胞上有甲状腺细胞特有的甲状腺球蛋白的表达.体外摄碘试验结果显示,用该方法培养的甲状腺细胞具备良好的摄碘功能,该功能具有对温度和高氯酸盐的双重敏感性.结论 建立了简便可行的小鼠甲状腺细胞的原代培养方法,为开展甲状腺细胞的摄碘及其他相关功能的研究奠定了基础.     

不同浓度雌二醇对甲状腺细胞增殖和凋亡的影响

目的研究不同浓度雌二醇(E2)对甲状腺细胞生长的调节作用。方法收集良性甲状腺肿瘤2例,男女各1例,恶性甲状腺肿瘤1例,女性,分别取肿瘤组织及瘤旁组织,分6组培养原代细胞,分别以10-11、10-10、10-9、10-8mol/LE2干预,用细胞计数及MTT法观察细胞生长情况;一部分细胞同样以10-11、10-10、10-9、10-8mol/LE2干预,加DMBA诱导凋亡,MTT观察凋亡情况。结果不同浓度E2干预后,可不同程度地增强甲状腺细胞增殖,多数有统计学差异(P<0.05);不同浓度的E2可不同程度地抑制甲状腺细胞凋亡。结论 E2对甲状腺正常细胞及良、恶性肿瘤细胞有促增殖、抑凋亡的作用。     

甲状腺过氧化物酶及其自身抗体检测的研究进展

甲状腺过氧化物酶及其自身抗体检测的研究进展徐荣佳综述武建国审校（南京军区南京总医院临床免疫科，南京２１０００２）甲状腺过氧化物酶（ＴｈｖｒｏｉｄＰｅｒｏｘｉｄａｓｅ，ＴＰＯ）是甲状腺微粒体（ＴｈｙｒｏｉｄＭｉｃｒｏｓｏｍａｌ，ＴＭ）的主要抗原成分［１...     

甲状腺过氧化物酶抗体及甲状腺功能对早孕妊娠结局的影响

目的探讨甲状腺过氧化物酶抗体及甲状腺功能对早孕流产的影响。方法选取422例早孕妇女(孕周6~12周),分为3组,A组:稽留流产组(148例);B组:先兆流产组(150例);C组:正常早孕组(124例)。检测甲状腺过氧化物酶抗体(TPOAb)及甲状腺功能(TSH、FT4、TT4、FT3、TT3)。结果 A组TPOAb水平较B组及C组增高,差异有统计学意义(P<0.05)。A组FT4水平较B组及C组降低,差异有统计学意义(P<0.05)。结论TPOAb水平增高及FT4水平减低可能是导致稽留流产的因素。更多还原     

血清甲状腺过氧化物酶抗体含量对甲状腺功能亢进放射性碘治疗疗效的影响

目的探讨治疗前血清甲状腺过氧化物酶抗体(TPOAb)含量对甲状腺功能亢进放射性碘治疗后疗效的影响。方法回顾性分析使用放射性碘治疗的甲状腺功能亢进患者219例[男46例,女173例,平均年龄(38.0±12.8)岁],治疗后随访6个月并进行疗效评价。按照疗效评价的结果将患者分为:无效组、好转组、治愈组、甲状腺功能减退(简称甲减)组。使用秩和检验、Logistic回归判断TPOAb含量在各组间的差异是否具有统计学意义,使用Spearman相关性检验分析TPOAb含量与疗效的关系,使用ROC曲线计算以TPOAb含量为指标预测患者预后的敏感度和特异度。结果 TPOAb的含量在四组间具有统计学差异(秩和检验:χ2=25.237,P=0.000;Logistic回归分析:B=-0.006,P=0.011),且随着TPOAb含量的升高患者出现无效及甲减的概率上升。以TPOAb<69.3 IU/ml作为诊断预后为治愈的指标,敏感度60.7%,特异度90.6%。结论治疗前血清TPOAb含量升高使放射性碘治疗后患者出现甲减及无效的概率增大,使用TPOAb<69.3 IU/ml作为判断预后治愈的敏感度、特异度较好。     

抗甲状腺球蛋白抗体与抗甲状腺过氧化物酶抗体在甲状腺功能异常诊断中的应用

目的观察抗甲状腺球蛋白抗体(A-TG)、抗甲状腺过氧化物酶抗体(A-TPO)与甲状腺功能异常的关系,探讨其在临床诊断中的应用价值。方法以电化学发光法检测140例甲状腺功能异常的患者(按病情分为亚临床甲亢组、临床甲亢组、亚临床甲减组与临床甲减组)和226例甲状腺功能正常者(对照组)的血清A-TG与A-TPO的水平。结果亚临床甲亢组、临床甲亢组、亚临床甲减组与临床甲减组的A-TG水平、A-TPO水平、A-TG、A-TPO阳性率均明显高于对照组,差异均有显著意义(P<0.01)。结论血清A-TG和A-TPO检测异常与甲状腺功能损伤间存在密切关系。血清A-TG和A-TPO可作为甲状腺功能异常的辅助诊断指标。     

甲状腺过氧化物酶抗体和孕妇甲状腺功能障碍的筛查

目的探讨产前孕妇甲状腺过氧化物酶抗体和促甲状腺素筛查的重要性。方法根据甲状腺功能测定结果,将孕妇分为甲状腺功能紊乱组(Ⅰ组)53例,非甲状腺功能紊乱组(Ⅱ组)49例,采用电化学方法检测孕妇血清TPO-Ab和甲状腺激素TSH水平。结果Ⅰ组TOP-Ab阳性率高达79.24%,Ⅱ组TOP-Ab阳性率高达10.20%;47例孕妇TPOAb阳性,其中有10例TSH异常(21.3%),55例TPOAb阴性,其中4例TSH异常(7.5%)。结论 TPOAb阳性孕妇甲状腺功能紊乱明显增加,TPOAb阳性孕妇的甲状腺功能有向亚临床甲状腺功能减退的趋势。     

甲状腺球蛋白抗体、微粒体抗体及过氧化物酶抗体检测在诊断甲状腺疾病中的价值

目的探讨甲状腺球蛋白抗体（TG-Ab）、甲状腺微粒体抗体（TM-Ab）、甲状腺过氧化物酶抗体（TPO-Ab）诊断甲状腺疾病的临床价值。方法应用电化学发光法检测92例甲状腺疾病及66例其他疾病患者血清TPO-Ab浓度,应用问接免疫荧光法检测TG-Ab及TM-Ab阳性率。结果三者阳性率在甲状腺疾病组与非甲状腺疾病组间差异有统计学意义（P〈0．01）;TPO-Ab诊断甲状腺疾病的特异性、准确性、阳性预测值及阳性似然比分别为93％、69．6％、90．7％及7．6;TPO-Ab与TM-Ab在甲状腺疾病组间呈显著相关（r=0．6,P〈0．01）。结论TPO-Ab在甲状腺疾病中的诊断价值相对较高;TG-Ab和TPO-Ab联合检测是自身免疫性甲状腺疾病诊断的有效组合。     

血清甲状腺过氧化物酶抗体检测在甲状腺功能紊乱中的临床探讨

甲状腺过氧化物酶是一种糖基化血红素蛋白．跨于甲状腺细胞顶缘的细胞膜上．伸向充满滤泡腔的部分具有催化活性,是甲状腺激素合成的关键酶,也是一个重要自身抗原,当其作为抗原刺激机体产生抗体时常引起自身免疫性甲状腺病。此情况下．甲状腺疾病患者有相当一部分人早期甲状腺激素正常或轻度变化．     

Studies on human iodide peroxidase: its activity in various thyroid disorders.

Iodide peroxidase activities in the human thyroid gland were measured in various thyroid disorders. The peroxidase activities in the thyroid gland revealed high values in Graves' disease, normal levels in thyroid adenoma and low values in thyroid carcinoma as compared to those in normal thyroid glands. A high activity of this enzyme was observed in toxic adenoma. The same results were observed whether 10% (W/V) homogenate or mitochondrial and microsomal fractions of the thyroid gland were used.     

Effects of sodium fluoride on total serum protein levels and transaminase activity in rats.

Transaminase activity and serum total protein level were investigated in adult rats after oral treating with sodium fluoride at three doses, 10, 20 and 30 mg/kg daily for 90 days. After 90 days, the average total serum protein level of the rats in the treatment group decreased significantly compared with that in the control [1.9 +/- 0.1 (mean +/- S.D., n = 140) vs. 3.1 +/- 0.2] mg/dl, P< 0.05. Serum transaminase activity in the treatment group increased compared with that in the control [5.3 +/- 0.4 (mean +/- S.D., n = 140) vs. 3.2 +/- 0.3] micromol/min per ml, P < 0.05.     

Effect of sodium fluoride, ampicillin, and chlorhexidine on Streptococcus mutans biofilm detachment

We examined the effect of three clinically used antimicrobials on Streptococcus mutans UA159 biofilm detachment under flow conditions. Sodium fluoride (NaF) and chlorhexidine at MIC levels promoted biofilm detachment and inhibited detachment when concentrations were higher than the MIC and reduced detached-cell viability only at high concentrations. Ampicillin at all concentrations tested inhibited detachment and reduced the percentage of viable biofilm-detached cells. All the three antimicrobial treatments reduced biofilm live/dead cell ratios.     

Improved method of thyroid peroxidase extraction from the human thyroid gland

This study describes a new method of solubilizing thyroid peroxidase (TPO) and partial purification of TPO from a small surgical specimen of human thyroid tissue. Graves' thyroid tissue was homogenized and centrifuged to obtain the 100 000 X g pellet. To solubilize TPO from the 100 000 X g pellet protein, the following four detergents were used: Triton X-100, digitonin, sodium deoxycholate, and 3-[(3-choramidpropyl)-dimethylammonio] 1-propanesulfate (CHAPS). For some samples, two detergents were combined and trypsin was also used. The best solubilization of TPO activity was obtained from the combination of digitonin-CHAPS-trypsin treatment or deoxycholate-CHAPS-trypsin treatment. The solubilized crude TPO was then chromatographed on a Sephacryl S 300 column. The results of chromatography indicated that detergent treatment alone did not separate TPO from other membrane proteins and the addition of trypsin was required for separation of TPO. Sephacryl chromatography of detergent-trypsin solubilized TPO was suitable as an initial step for purification of TPO from a small human thyroid tissue.     

Differential activation of rabbit femoral arteries by aluminum fluoride and sodium fluoride

The effect of fluoride (NaF; 10 mM sodium fluoride plus deferoxamine to chelate contaminating aluminum) and fluoride plus aluminum fluorides (AlF; 10 mM sodium fluoride plus 20 microM aluminum chloride) on activation of rabbit femoral arteries was investigated. AlF and NaF produced large increases in stress (force/muscle cross-sectional area), but temporal changes were dissimilar, as were other indices of muscle activation. Stress produced by NaF developed slowly and only after a long delay of about 15 min, whereas stress produced by AlF developed rapidly after a delay of only about 5 min. NaF-induced contractions were more sustained than AlF-induced contractions. Both AlF and NaF increased the level of cross-bridge phosphorylation and the velocity of muscle shortening, but at comparable stresses, AlF produced greater increases than did NaF. AlF produced a large increase in lP production, whereas NaF produced a small increase. Also, AlF-induced stress was largely insensitive to inhibition by the calcium channel blocker, nifedipine (1 microM), whereas NaF-induced stress was largely inhibited by nifedipine. However, in tissues depleted of calcium, both agents produced potent contractions when CaCl2 was added back to the tissues (EC50 values for AlF, NaF, histamine, phenylephrine and KCl were, respectively, 0.057, 0.085, 0.11, 0.11 and 0.23 mM). AlF, but not NaF, strongly desensitized arteries to phenylephrine, causing a 73% reduction in the ability of phenylephrine to achieve maximum steady-state stress. These data suggest that fluoride contracted rabbit femoral arteries by stimulating L-type calcium channels, and that aluminum fluoride stimulated phospholipase C, producing additional muscle activation.     

氟对成纤维细胞核心结合因子α1及增殖活性量-效关系的影响

背景:氟通过刺激成纤维细胞向成骨细胞方向转化,可能在氟性骨损伤骨周化骨中起重要作用。核心结合因子α1作为成骨细胞特异性转录因子,是成骨细胞分化以及成骨的必要条件。目的:观察不同质量浓度氟化钠干预体外培养新生大鼠皮肤成纤维细胞不同时间后核心结合因子α1的表达及成纤维细胞的增殖活性。方法:采用细胞原代培养方法,将细胞分为对照组和7个染氟组(0.0001,0.001,0.01,0.1,1,10,20mg/L),分别在4个染氟时间段(24,48,72,96h)收集细胞培养上清液,应用酶联免疫法测定核心结合因子α1的含量;采用四甲基偶氮唑盐比色法观察氟对成纤维细胞增殖活性的影响。结果与结论:氟能提高成纤维细胞核心结合因子α1的表达,核心结合因子α1可能在成纤维细胞致氟性骨损伤骨周化骨中起重要作用。氟对成纤维细胞的增殖活性的影响呈一定的剂量-时间-效应关系,短时间-低剂量的氟可提高成纤维细胞的增殖活性,随着染氟剂量的增加及染氟时间的延长,成纤维细胞增殖活性明显减弱。     

Human thyroid peroxidase: Autoantibody recognition depends on the natural conformation

Thyroid peroxidase (TPO) purified from human thyroid glands was used to study the correlation between natural conformation and its ability for binding of autoantibodies in sera from patients with autoimmune lymphocytic thyroiditis (Hashimoto's thyroiditis). Treatment with heat (60°C), urea, sodium dodecylsulfate (SDS), and dithiothreitol (DTT) decreased the autoantigenicity of TPO on average by 73.3%, 74.1%, 86.3%, and 91.8% measured by enzyme-linked immunosorbent assay (ELISA), respectively. These data demonstrate, that the binding of the majority of autoantibodies to human TPO depends on its natural conformation and modification. © 1993 Wiley-Liss, Inc.