雷帕霉素纳米粒滴眼液联合环孢素A缓释膜治疗兔高危角膜移植术后免疫排斥反应的研究

Objective To evaluate the combined effect of topical rapamycin (RAPA) eye drop in nanometer vector and poly (lactic acid) (PLA) wafers of cyclospoirne A(CsA) in the prevention of acute allograft rejection after rabbit corneal transplantation. Methods It was an experimental study. RAPA was incorporated into the nanometer particles and CsA was incorporated into PLA wafers. A was syngeneic control whose both donor and recipient are New Zealand rabbit. Gray donor corneas were implanted into the 102 recipients of New Zealand albino rabbits with corneal neovascularization who were randomly divided into B, C, D, E, F, G 6 groups to receive the different types of therapy: B was no therapy control; C was eye drop of nanometer vector but no RAPA twice a day,28 days; D was PLA wafers in the anterior chamber of rabbit eyes but no drugs; E was 0.5% RAPA eye drop of nanometer vector twice a day,28 days; F was PLA wafers of CsA in the anterior chamber of rabbit eyes; G was PLA wafers of CsA in the anterior chamber of rabbit eyes and 0.5% RAPA eye drop of nanometer vector eye drop twice a day for 28 days together. Postoperative evaluation included slit-lamp biomicroscopy, histopathology and immunohistoiogy, Cytokines related with neovascularization and immunosuppression in the corneal tissue by RT-PCR. The graft survival was assessed by One-Way ANOVA and q test. Results Corneal allograft survival time: A (100.00±0.00), B ( 8.44±1.24), C (8.89±2.57), D (8.56±2.30), E (43.11±5.58), F (43.67±9.54), G (72.00±15.34)d. Group G led to a statistically significant prolongation of transplant survival and was superior than group E and F which was a statistical prolongation compared with group B, C and D (qGE=11.42, qGF=11.24,qEB=13.64, qEC=13.38, q<=13.46, qFB=13. 82, qFC=13.56, qFD=13.64; P<0.01). Immunohistopathologically, the grafts were subjected to an immune response contained a dense infiltrate of neutorphils,CD4+ and CD8+ T lymphocytes in the group B ,C and D. This cellular infiltrate was a significant reduction in group E,F,G. RT-PCR showed that the gene expression of IL-2 was inhibited earlier (3 days) in group F, G and VEGF gene expression being suppressed later (14 days) in group E, G. Conclusions Combined therapy with topical application of RAPA eye drop of nanometer vector and CsA PLA wafers can significantly prolong the survival of allograft at high-risk. Moreover, topical combined treatment of them is more effective, lower dosage, less side-effects and cheaper than the treatment with topical individual immunosuppressive drug.     

碘/钠摄入量对小鼠血脂代谢影响的实验研究

Objective The present study has been designed to investigate the impact of dietary iodine/sodium intake on blood lipid metabolism in mice. Methods According to body weight and gender, two hundred and sixty Balb/c mice were randomly divided into 2 groups including normal sodium group(Na) and low sodium group(LNa), with 130 animals per group. Each group were then randomly further divided into 5 sub-groups according to the amount of iodine intake: ① severe iodine deficiency(SID); ② mild iodine deficiency(MID); (③normal iodine (NI); ④ 10-fold high iodine ( 10HI ); (⑤ 50-fold high iodine (50HI), 10 groups in total, 26 per group.Eight months later, the body weight and the levels of urinary iodine, thyroid hormones and total cholesterol (TC),Results In Na group, the levels of TG and TC in male mice of SID group[ (1.64 ± 0.35), (3.88 ± 0.35 )mmol/L]and MID group[ ( 1.67 ± 0.31 ), (3.41 ± 0.66)mmol/L] were significantly higher than that of NI group[ ( 1.49 ± 0.42), (3.25 ± 0.47)mmol/L] and the levels of TG in female mice of SID group[(1.52 ± 0.22)mmol/L] were significantly higher than that of NI group[ (1.23 ± 0.22)mmol/L]. In addition, the levels of TG in male mice of 10HI and 50HI groups [ ( 1.16 ± 0.23 ), ( 1.21 ± 0.27 ) mmol/L ] were significantly lower than that of NI group [ ( 1.49 ± 0.42)mmol/L, all P ＜ 0.05], the levels of TC in female mice of 10HI and 50HI groups[(2.37 ± 0.49), (2.48 ± 0.37)mmol/L] were significantly lower than that of NI group[ (2.84 ± 0.37) mmol/L, all P ＜ 0.05 ]. In LNa group,the levels of TG and TC in male mice of SID group[ (1.39 ± 0.40), (3.33 ± 0.46 )mmol/L] were significantly lower than that of NI group [(1.30 ± 0.28), (3.00 ± 0.53) mmol/L, all P ＜ 0.05], the levels of TG, TC and LDL in female mice of SID group[ (1.48 ± 0.26), (2.76 ± 0.43), (0.62 ± 0.22)mmol/L], the levels of LDL in female mice of MID group[ (0.60 ± 0.17 )mmol/L] were significantly lower than that of NI group[(l.22 ± 0.36), (2.51 ± 0.38),(0.48 ± 0.08), (0.48 ± 0.08)mmol/L, all P ＜ 0.05], the levels of TG in male mice of 10HI and 50HI group [ (1.12 ± 0.22), (0.90 ± 0.11 )mmol/L] were significantly lower than that of NI group (all P ＜ 0.05 ), the levels of TC in female mice of 10HI and 50HI groups[ (2.35 ± 0.34), (2.37 ± 0.37)mmol/L], the levels of LDL in female mice of 50HI group[(0.65 ± 0.18)mmol/L], were significantly lower than that of NI group(all P ＜ 0.05). In Na group, the levels of thyroid hormones were distinctively decreased in SID group[TT4(0.00 ± 0.00)nmol/L, FT4 (0.93 ± 0.42)pmol/L, TT3(0.49 ± 0.07)nmol/L, FT3(2.86 ± 0.37)pmol/L] and MID group [TT4 (17.15 ± 15.26)nmol/L, FT4( 18.46 ± 4.31 )pmol/L, TT3(0.67 ± 0. 10)nmol/L, FT3(3.18 ± 0.24)pmol/L] compared with that of the NI group [TT4 (37.15 ± 15.26)nmol/L, FT4(28.46 ± 4.31)pmol/L, TT3(0.85 ± 0.10)pmol/L, FT3(3.87 ± 0.24)pmol/L, all P ＜ 0.05 ]. In LNa group, the levels of thyroid hormones were distinctively decreased in SID group [TT4 (0.00 ± 0.00) nmol/L,FT4(1.03 ± 0.78)pmol/L, TT3(0.51 ± 0.05)nmol/L, FT3(3.01 ± 0.17)pmol/L] and MID group[TT4(19.76 ± 12.22)nmol/L, FT4(21.46 ± 5.37)pmol/L, TT3(0.71 ± 0.21)nmol/L, FT3(3.56 ± 0.23)pmol/L] compared with that of the NI group[TT4(36.23 ± 14.72)nmol/L, FT4(30.96 ± 6.33)pmol/L, TT3(0.89 ± 0.20)nmol/L, FT3(4.05 ± 0.24)pmol/L, all P ＜ 0.05]. Conclusions Dietary iodine intake plays an important role in the blood lipid metabolism. Iodine deficiency could increase while iodine excess could decrease the levels of serum TG, TC or LDL in mice. Monitoring the amount of iodine intake during sodium restriction should have an important role in effective prevention and treatment of cardiovascular disease.     

碘对诱发实验性自身免疫性甲状腺炎形成的影响

目的建立诱发型实验性自身免疫性甲状腺炎(EAT)动物模型,研究碘对诱发EAT形成的影响,进一步探讨其可能的作用机制。方法:采用雌性Lewis大鼠诱发EAT并饲以不同碘含量的饮食,观察各组大鼠甲状腺病理改变、血清中Tg自身抗体水平、血清中甲状腺激素水平、脾细胞分泌TNF水平和甲状腺FasmRNA表达水平。结果1.诱发EAT的HI组大鼠炎症范围较广,为较重度炎症。NI组显示为轻度炎症,且为恢复期。LI组显示为滤泡内少量淋巴细胞浸润。2.诱发EAT的HI、NI和LI组大鼠血清中均出现较高水平的Tg自身抗体,3组间无显著性差异。3.诱发EAT的HI组大鼠脾细胞分泌TNE水平高于NI和LI组(P<0.05)。4.诱发EAT的HI、NI和LI组大鼠甲状腺FasmRNA的表达水平均较各自未诱发EAT的大鼠明显升高,其中HI组高于同组的NI和LI组,P<0.01。结论1.高碘饮食可使已诱发EAT的Lewis大鼠甲状腺组织炎症加重且炎症反应持续时间较长,考虑与脾细胞分泌的TNF水平和甲状腺FasmRNA的表达水平较高有关。2.未发现甲状腺组织炎症的程度与自身抗体水平相关。     

低碘后补充不同碘水平饮食对大鼠肿瘤坏死因子和干扰素mRNA表达的影响

目的大鼠低碘后,补充不同碘水平饮食,观察其甲状腺组织形态、甲状腺激素水平和脾脏中肿瘤坏死因子(TNF-α)和干扰素(IFN-γ)的mRNA表达,探讨低碘后补碘对大鼠的影响。方法选用Wistar大鼠,低碘饲料加去离子水饲养3个月为低碘组(LI组),低碘后补充适碘饮食为LI-1×KI组,补充高碘饮食为LI-50×KI组,均补碘3个月,另设适碘对照组(NI组)。检测4组大鼠血清中甲状腺激素水平、甲状腺组织形态、脾脏中TNF-α和IFN-γmRNA的表达。结果LI组大鼠血清中T3和T4水平下降,甲状腺滤泡内胶质稀少,甲状腺上皮细胞呈增生、肥大等典型的低碘表现,脾脏中TNF-α和IFN-γ的mRNA表达也明显低于NI组。低碘后补充适碘饮食的LI-1×KI组,不仅使血清中T3和T4水平和甲状腺上皮细胞的形态逐渐恢复正常,TNF-α和IFN-γ的mRNA表达也明显提高。而低碘后补充高碘饮食的LI-50×KI组,甲状腺激素水平和TNF-α和IFN-γ的mRNA表达也均明显上升,但甲状腺组织形态虽有所好转,与LI-1×KI组比较尚有明显差异,且甲状腺滤泡有胶质潴留。结论低碘后补充适当水平的碘,其甲状腺激素水平、TNF-α和IFN-γ的mRNA表达及甲状腺组织形态的改变可逐渐恢复正常,呈可逆性。而低碘后补充高碘饮食,其甲状腺组织形态学的改变尚难恢复为正常。     

雌激素与自身免疫性疾病

自身免疫性疾病的发病存在明显的性别差异，女性的发病率常高于男性，文章就雌激素在自身免疫性疾病发生机制中的作用，特别是近年来雌激素对免疫细胞信号，细胞分化，下丘脑－垂体－肾上腺（HPA）轴的激活，自身免疫性疾病的发生和缓解等方面的研究作一介绍，进一步探讨雌激素调节异常对自身免疫性疾病发生的影响。     

雷帕霉素纳米粒滴眼液联合环孢素A缓释膜治疗兔高危角膜移植术后免疫排斥反应的研究

Objective To evaluate the combined effect of topical rapamycin (RAPA) eye drop in nanometer vector and poly (lactic acid) (PLA) wafers of cyclospoirne A(CsA) in the prevention of acute allograft rejection after rabbit corneal transplantation. Methods It was an experimental study. RAPA was incorporated into the nanometer particles and CsA was incorporated into PLA wafers. A was syngeneic control whose both donor and recipient are New Zealand rabbit. Gray donor corneas were implanted into the 102 recipients of New Zealand albino rabbits with corneal neovascularization who were randomly divided into B, C, D, E, F, G 6 groups to receive the different types of therapy: B was no therapy control; C was eye drop of nanometer vector but no RAPA twice a day,28 days; D was PLA wafers in the anterior chamber of rabbit eyes but no drugs; E was 0.5% RAPA eye drop of nanometer vector twice a day,28 days; F was PLA wafers of CsA in the anterior chamber of rabbit eyes; G was PLA wafers of CsA in the anterior chamber of rabbit eyes and 0.5% RAPA eye drop of nanometer vector eye drop twice a day for 28 days together. Postoperative evaluation included slit-lamp biomicroscopy, histopathology and immunohistoiogy, Cytokines related with neovascularization and immunosuppression in the corneal tissue by RT-PCR. The graft survival was assessed by One-Way ANOVA and q test. Results Corneal allograft survival time: A (100.00±0.00), B ( 8.44±1.24), C (8.89±2.57), D (8.56±2.30), E (43.11±5.58), F (43.67±9.54), G (72.00±15.34)d. Group G led to a statistically significant prolongation of transplant survival and was superior than group E and F which was a statistical prolongation compared with group B, C and D (qGE=11.42, qGF=11.24,qEB=13.64, qEC=13.38, q<=13.46, qFB=13. 82, qFC=13.56, qFD=13.64; P<0.01). Immunohistopathologically, the grafts were subjected to an immune response contained a dense infiltrate of neutorphils,CD4+ and CD8+ T lymphocytes in the group B ,C and D. This cellular infiltrate was a significant reduction in group E,F,G. RT-PCR showed that the gene expression of IL-2 was inhibited earlier (3 days) in group F, G and VEGF gene expression being suppressed later (14 days) in group E, G. Conclusions Combined therapy with topical application of RAPA eye drop of nanometer vector and CsA PLA wafers can significantly prolong the survival of allograft at high-risk. Moreover, topical combined treatment of them is more effective, lower dosage, less side-effects and cheaper than the treatment with topical individual immunosuppressive drug.     

抗甲状腺刺激性免疫球蛋白单链抗体基因的构建及序列测定

 目的构建抗甲状腺刺激性免疫球蛋白(Thyroid stimulating immunoglobulin,TSI)单链抗体基因,获得基因序列.方法利用噬菌体表面呈现技术,结合间接ELISA方法和竞争抑制实验,筛选出抗TSI单链噬菌体抗体,进一步获得抗TSI单链抗体,对抗TSI单链抗体基因进行测序.结果抗TSI单链抗体基因长759bp,其中VL长342bp,VH长372bp,具有开放性阅读框架,为功能性基因.结论抗TSI单链抗体基因构建成功,噬菌体表面呈现技术为获得功能性单链抗体基因的可靠方法.     

产后甲状腺炎发病机制的研究进展

产后甲状腺炎是发生于产后一年内的一种暂时性或永久性甲状腺功能紊乱的综合征,属于自身免疫性疾病.甲状腺超氧化物酶抗体与产后甲状腺炎的发生密切相关,目前认为,妊娠相关的免疫抑制在产后消失,出现暂时性免疫反弹是引起产后甲状腺炎的主要因素.此外,神经-内分泌-免疫网络调节、环境因素及遗传因素等的相互作用,决定了产后甲状腺炎的发生、发展.     

持久稳定分泌抗人甲状腺球蛋白单克隆抗体的杂交瘤细胞株的建立

杂交瘤技术创立以来,在生物、医学各领域已广泛应用,各地陆续报导建立了分泌抗白细胞抗原、肿瘤抗原和微生物抗原的单克隆抗体(McAb)杂交瘤细胞株。但在内分泌免疫病方面,有关报导还不多。内分泌免疫病是免疫介导性疾病中尤为复杂的一组疾病,单克隆抗体技术为诊断和深入细致研究其发病机制提供了有力工具。我们选择了     

自身免疫性甲状腺疾病患者血清瘦素水平观察

目的 观察自身免疫性甲状腺疾病(AITD)中Graves病(GD)和桥本氏甲状腺炎(HT)患者血清瘦素水平,探讨瘦素在AITD发病机制中的免疫学作用.方法 102例体质量指数(BMI)和年龄等因素相匹配的女性AITD初诊患者,根据临床表现及实验室检查结果分为3组:GD甲亢组,HT甲低组,亚甲低组,另设性别、年龄、BMI匹配的对照组.免疫化学发光法检测血清FT3、FT4、sTSH,ELISA法检测血清瘦素水平.结果 GD甲亢组血清FT3、FT4水平[(19.74±15.39)、(78.25±58.68)pmol/L]明显高于对照组[(4.87±0.25)、(15.96±3.15)pmol/L],而sTSH[(0.15±0.08)mU/L]和瘦素水平[(8.73±1.92)μg/L]明显低于对照组[(3.81±0.19)mU/L、(12.38±3.51)μg/L].组间比较差异均有统计学意义(P＜0.01或＜0.05).而HT甲低组兀FT3、FT4水平[(3.36±0.26)、(6.95±3.29)pmol/L]均明显低于对照组(P＜0.05),而sTSH[45.48±35.83)mU/L]和瘦素水平[(17.17±3.82)μg/L]明显高于对照组(P＜0.01或＜0.05).亚甲低组FT3、FT4水平[(4.67±0.60)、(14.87±2.14)pmol/L]与对照组比较差异无统计学意义(P＞0.05),而sTSH[(13.67土8.66)mU/L]和瘦素水平[(16.25±3.67)μg/L]均明显高于对照组(P＜0.01或＜0.05).结论 瘦素在AITD发病机制中可能具有一定的免疫调节作用,而AITD患者瘦素水平也可能受sTSH影响.