LHRH融合蛋白对雄性大鼠生殖功能的影响

目的 :以促黄体生成素释放激素融合蛋白 (Trx LHRH)为免疫疫苗 ,氢氧化铝为佐剂 ,研究Trx LHRH诱导雄性大鼠抗体生成及其对大鼠生殖功能的影响。　方法 :Trx LHRH为体外克隆融合基因在大肠埃希菌BL2 1表达的融合蛋白。以Trx LHRH为免疫疫苗 (0 .2mg)、氢氧化铝为佐剂 ,通过初次和加强免疫雄性成年SD大鼠 ,间接酶联免疫法测定抗体滴度 ,放射免疫法测定血清睾酮 (T)水平 ,同时观察附睾精子数量的变化。　结果 :Trx LHRH免疫 4周后抗体滴度达 1∶12 80～ 1∶2 5 6 0 ,加强免疫后直至 6周抗体滴度仍保持 1∶2 0 0 0高水平。血清T水平则显著降低 (P <0 .0 1) ,个体之间存在一定的差异 ;精子计数减少 (P <0 .0 5 )。　结论 :Trx LHRH能成功诱导大鼠产生抗LHRH抗体 ,并进而抑制垂体 睾丸轴的功能 ,抑制其生殖功能。     

NR2B多肽疫苗对神经病理性痛大鼠的镇痛效应

目的 评价NR2B多肽疫苗对神经病理性痛大鼠的镇痛效应.方法 成年雌性SD大鼠,体重180～200 g,制备保留性神经损伤(SNI)模型,7 d后选取SNI模型制备成功的大鼠18只,随机分为3组(n=6),NR2B组沿背部两侧皮下多点注射NR2B多肽疫苗50 μl(含NR2B 100μg)3次,每隔2周注射1次;PBS组和KLH组分别给予PBS 50 μl和匙孔虫戚血蓝蛋白100 μg,均用弗氏佐剂稀释至100 μl.分别于制备SNI前1 d(基础值)、第1次免疫前1 d及第3次免疫后14d时测定机械痛阈;分别于制备SNI前1 d及第3次免疫后14 d时取血清和脑脊液,测定NR2B抗体滴度,然后处死大鼠,取L3-5脊髓组织,其中3只采用免疫组化法测定脊髓背角胶质纤维酸性蛋白表达水平,反映星形胶质细胞激活程度;另外3只采用Western blotting法测定脊髓背角NR2B蛋白表达水平.结果 NR2B组第3次免疫后14 d时血清及脑脊液中均可检测到NR2B抗体,而SNI前1 d时未检测到NR2B抗体,PBS组及KLH组各时点均未检测到NR2B抗体.与基础值比较,各组机械痛阈均降低(P<0.05);与PBS组和KLH组比较,NR2B组第3次免疫后机械痛阈升高,脊髓背角星形胶质细胞激活程度降低,NR2B蛋白表达下调(P<0.05).结论 NR2B多肽疫苗对神经病理性痛大鼠具有镇痛作用,其机制与进入脊髓的NR2B抗体下调了NR2B蛋白表达有关.     

Nur77在不同发育阶段小鼠睾丸的表达及其功能

目的观察核受体Nur77在不同发育阶段小鼠睾丸的表达,并探讨其对睾酮合成的作用。方法用1周龄的ICR雄性小鼠作为幼年期组(相当于人类幼年期,n=6),4周龄的ICR雄性小鼠作为性发育前组(相当于人类青春期前,n=6),12周龄的ICR雄性小鼠作为性成熟期组(相当于人类青壮年,n=6),12月龄的ICR雄性小鼠作为老年组(相当于人类中老年,n=6)。Real time PCR和Western Blot分别检测小鼠睾丸中Nur77 mRNA和蛋白水平表达。体外培养小鼠睾丸间质细胞系MLTC-1,Nur77干涉腺病毒(Ad-siNur77)及空载腺病毒(Ad-CMV)转染MITC-1。放射免疫法检测MLTC-1细胞培养液上清的睾酮含量,Real time pCR和Western Blot检测StAR及HSD3B的表达。结果 Nur77的mRNA水平在青春期前表达量最高,蛋白水平则在性成熟期小鼠睾丸中最高;Nur77干涉后MLTC-1细胞的睾酮合成含量较末干涉组明显降低;Nur77干涉后MLTC-1细胞的StAR和HSD3B的表达较未干涉组明显降低。结论 Nur77在小鼠睾丸组织的表达随年龄变化而变化,这种变化对于维持小鼠体内的睾酮水平可能有重要意义。     

反复注射不同剂量氯胺酮对幼年小鼠认知功能的影响

目的 探讨反复注射不同剂量氯胺酮对幼年小鼠认知功能的影响.方法 21 d龄小鼠50只,体重10～15 g,随机分为5组(n=10),正常对照组(c组);生理盐水组(N组)腹腔注射生理盐水0.1 ml,1次/d,连续7 d;不同剂量氯胺酮组(K1组、K2组、K3组)分别腹腔注射氯胺酮25、50、100 mg/kg,1次/d,连续7 d.于给药结束后1 d时测定学习功能,于给药结束后2 d时测定记忆功能.记忆功能测定结束后每组处死4只小鼠,采用免疫组化法测定海马脑源性神经营养因子(BDNF)表达;处死6只小鼠,采用免疫印记法测定海马BDNF蛋白表达.结果 与C组比较,N组和K1组学习记忆功能减退,BDNF蛋白表达下调(P＜0.05或0.01),K2组和K3组差异无统计学意义(P＞0.05);N组和K1组学习记忆功能和BDNF蛋白表达差异无统计学意义(P＞0.05).结论 50、100 mg/kg(连续7 d反复给药)氯胺酮对幼年小鼠认知功能无影响,25 mg/kg(连续7 d反复给药)氯胺酮可降低认知功能.     

19-去甲睾酮对性腺发育不良小鼠附性腺生长的影响

目的 :观察合成同化甾体激素 19 去甲睾酮 (NT)对性腺发育不良 (hpg)小鼠的前列腺、精囊腺 (SV)、附睾生长发育的影响 ,为用药的安全性提供实验依据。　方法 :NT硅胶囊皮下埋植 5周缓释给药 ,设给药hpg小鼠实验组 (n =7)、不给药hpg小鼠对照组 (n =7)和性腺发育正常小鼠对照组 (n =10 )。给药结束后测定前列腺腹侧叶(VP)、SV和附睾的重量及VP腺管末梢数目。　结果 :与hpg小鼠对照组相比 ,hpg小鼠给药组各附性腺的重量均显著增加 (P <0 .0 0 5 ) ,VP腺管分支形态发育趋于正常 ;hpg小鼠给药组SV的重量与正常小鼠相同 ,但VP、附睾的重量及VP腺管末梢数目仍显著低于正常性腺小鼠对照组 (P <0 .0 0 5 )。　结论 :NT能明显刺激hpg小鼠附性腺的生长发育     

HMGB1在肠缺血再灌注小鼠肺损伤中的作用：与NETs的关系

目的评价高迁移率族蛋白B1(HMGB1)在肠缺血再灌注小鼠肺损伤中的作用及其与中性粒细胞胞外诱捕网(NETs)的关系。方法 SPF级健康成年雄性C57BL/6小鼠24只,体重20～25 g,采用随机数字表法分为4组(n=6):假手术组(Sham组)、肠缺血再灌注组(I/R组)、IgY对照抗体组(IgY组)和Anti-HMGB1中和抗体组(Anti-HMGB1组)。采用夹闭肠系膜上动脉1 h后恢复灌注的方法制备小鼠肠缺血再灌注损伤模型。Anti-HMGB1组于缺血1 h后肠道血流恢复即刻腹腔注射Anti-HMGB1中和抗体1.0 mg/kg;IgY组于缺血1 h后肠道血流恢复即刻腹腔注射IgY同型对照抗体1.0 mg/kg。于再灌注4 h时处死小鼠,收集小鼠支气管肺泡灌洗液(BALF),测定总蛋白、游离双链DNA(dsDNA)及HMGB1的浓度;取肺组织HE染色后光镜下观察病理学结果,采用Western blot法测定瓜氨酸化组蛋白H3(Cit-H3)表达。结果与Sham组比较,I/R组和IgY组肺组织病理学损伤评分、BALF总蛋白、HMGB1和dsDNA浓度升高,Cit-H3表达上调(...     

雄激素对男性性腺功能低减病人促红细胞生成素的影响

目的 :观察雄激素替代治疗对性腺功能低减男子促红细胞生成素 (EPO)的影响 ,探讨雄激素促进红细胞和血红蛋白生成增加的机制。　方法 :8例原发性性腺功能低减 (Klinefelter综合征 )病人 ,接受初次 5 0 0mg或10 0 0mg十一酸睾酮 (TU)肌肉注射 ,间隔 3个月后交叉剂量第二次注射。注射前后观察血清性激素水平的变化 (放免法 ) ;在注射前和后的第 4、8周分别查血常规和红细胞比容 ,酶免疫法测定血清EPO。　结果 :注射TU后 ,第二性征发育情况改善 ,血睾酮水平显著升高 ,注射后 1周达峰值 ,维持有效治疗的雄激素水平 (>10nmol/L)超过 6周。血红细胞计数、红细胞比容和血红蛋白含量的均数均有不同程度的增加趋势 ,但统计学上差异不显著 (P >0 .0 5 )。与治疗前相比 ,注射TU后EPO水平显著升高 ,并维持 8周以上 (P <0 .0 1～ 0 .0 5 ) ;第 2次注射TU仍然使EPO水平升高。　结论 :雄激素替代治疗使性腺功能低减男子EPO水平升高 ,是红细胞生成增加的机制之一。     

脊髓mTOR/S6K1/Gli1信号通路在小鼠慢性吗啡耐受中的作用

目的评价脊髓哺乳动物雷帕霉素靶蛋白(mTOR)/核糖体蛋白S6激酶1(S6K1)/Gli1信号通路在小鼠慢性吗啡耐受中的作用。方法健康雄性昆明小鼠,8～10周龄,体重23～25 g。实验Ⅰ取小鼠50只,采用随机数字表法分为2组:生理盐水组(S组,n=10)和吗啡组(M组,n=40)。M组和S组分别皮下注射吗啡和生理盐水10 mg/kg,2次/d,连续7 d。于给药前1 d和每天最后1次给药后30 min测定热痛阈,计算最大镇痛效应百分比(MPE)。M组于给药后1、3、5和7 d热痛阈测定结束后、S组于最后1次热痛阈测定结束后随机处死10只小鼠,取脊髓组织。实验Ⅱ取小鼠40只,采用随机数字表法分为4组(n=10):KU-0063794+吗啡组(KU+M组)、二甲基亚砜(DMSO)+吗啡组(DM+M组)、吗啡+KU-0063794组(M+KU组)和吗啡+二甲基亚砜组(M+DM组)。4组皮下注射吗啡10 mg/kg,2次/d,连续7 d。于第1～3天每天吗啡注射前30 min时,KU+M组腹腔注射mTOR特异性抑制剂KU-0063794 200 μl(1 μg/μl),DM+M组腹腔注射10...     

PEI增强G250 DNA疫苗和重组肽疫苗的prime-boost免疫策略研究

目的 :探讨PEI作为免疫佐剂对G250抗原肽基因PVAX1/C-G250肽-C免疫保护效果的增强作用及联合应用CAIX蛋白疫苗进行PRIME—BOOST免疫程序免疫增强效果。方法:用Eco R I、Xho I和Eco R I、Sal I分别双酶切PVAX1及既往构建的p ET28a(+)/C-G250肽-C质粒。利用DNA重组技术构建重组质粒PVAX1/C-G250肽-C,酶切分析鉴定。大量提取质粒并分光光度计测质粒含量。将32只雌性昆明小鼠随机分为(A)裸DNA组,(B)DNA-PEI复合物组,(C)DNA-PEI+蛋白疫苗组,(D)空白对照组。按0,10,20,30天程序经股四头肌注射免疫。C组在第20天及第30天进行蛋白冲击。初次免疫前和第40天鼠尾取血,ELISA法检测抗体滴度。流式细胞术测淋巴细胞亚群CD4+和CD8+。结果:酶切及基因测序鉴定证实G250抗原肽c DNA正确插入PVAX1/C-G250肽-C真核表达的重组质粒中。昆明小鼠经4次免疫后,3个实验组都产生了特异性的体液和细胞免疫反应,B组的抗体滴度1:1.28×104及CD4+、CD8+达26.12%和12.60%,明显高于A组的1:3.2×103和CD4+,CD8+占到19.32%和10.74%。而蛋白冲击组的1:5.12×104和CD4+,CD8+占到41.96%和15.14%,明显高于B组(P0.05)。结论:成功构建重组质粒PVAX1/C-G250肽-C。该DNA疫苗与PEI和G250蛋白疫苗联合使用后产生极强的免疫原性,诱导产生了高滴度、高特异性抗体及细胞免疫反应。证实G250DNA疫苗联合PEI使用并进行蛋白疫苗冲击的免疫策略可产生强大的免疫保护作用。为恶性肿瘤的术后辅助治疗提供新的思路和方法。     

雷帕霉素联合IL-2融合蛋白在小鼠皮肤移植模型中对乙肝疫苗免疫原性的影响

目的研究雷帕霉素联合IL-2融合蛋白对小鼠皮肤移植术后乙肝疫苗免疫原性的影响,寻求移植术后预防乙型肝炎的有效方法。方法 DBA小鼠为供体,C57BL/6小鼠为受体,行皮肤移植后分为联合治疗组、雷帕霉素组、疫苗组,每组10只,联合治疗组给予雷帕霉素(1mg/kg,腹腔内注射)+IL-2融合蛋白(1μg/只,腹腔内注射)+疫苗;雷帕霉素组只给予雷帕霉素+疫苗;疫苗组仅给予乙肝疫苗。以上3组均在皮肤移植后第一天给予乙肝疫苗(2μg/只,肌肉注射)。空白组不给于任何处理。14d后测定各组血清中乙肝抗体滴度水平、IL-4、IL-10、IL-2、IFN-γ水平,应用RT-PCR测定各组淋巴组织中IL-21、FoxP3及Bcl-6的表达水平,同时随访小鼠皮肤移植物存活时间。结果联合治疗组移植术后乙肝表面抗体(HBsAb)滴度明显高于雷帕霉素组(P<0.05),皮肤移植物的存活时间也明显延长(P<0.05);联合治疗组Th2类因子IL-4,IL-10的水平明显低于雷帕霉素组(P<0.05),而Th1类因子IL-2和IFN-γ水平明显较高(P<0.05)。联合治疗组混合淋巴细胞中IL-21、FOXP3的表达水平明显高于雷帕霉素组(P<0.05),而Bcl-6的表达水平明显较低(P<0.05)。结论雷帕霉素联合IL-2融合蛋白联合应用能够在移植术后增强乙肝疫苗产生抗体的滴度,同时延长移植物的存活时间,为临床移植术后提高疫苗免疫原性提供了一种治疗策略。     

免疫增强型胰腺癌MUC1-VNTR-C144 DNA疫苗的构建

目的 构建免疫增强型胰腺癌MUC1-VNTR-C144 DNA疫苗.方法 在PeDNA 3.1-VNTR/Myc-his(+)A质粒靶基因之后插入乙型肝炎病毒核心抗原C144基因,构建MUC1-VNTR-C144 DNA疫苗.45只C57BL/6小鼠随机分3组,VC组接种PeDNA 3.1-VNTR-C144/Myc-his(+)A质粒,V组单纯接种PeDNA 3.1-VNTR/Myc-his(+)A质粒,C组单纯接种pcDNA3.1-C144/Myc-his(+)A质粒.4周后加强免疫1次.结果 VC组小鼠脾细胞毒性T淋巴细胞(CTL)的特异性杀伤率(46.7±8.2)%显著高于V组(34.8±3.1)%和C组(12.9±1.2)%(E/T=80/1,P<0.01).而CTL对未经VNTR合成肽孵育的靶细胞EIA-VNTR杀伤率较低(P<0.01),VU 3C6可抑制CTL对经VNTR合成肽孵育的靶细胞EIA-VNTR+的杀伤活性(P<0.01),表明具有VC组诱生的CTL应答依然保持VNTR特异性.VC组小鼠血清抗VNTR抗体等效浓度(2810.2±308.3)mg/L高于V组(2323.5±238.3)mg/L和C组(2130.9±138.5)mg/L,差异有统计学意义(P<0.01).结论 增强型胰腺癌MUC1-VNTR-C144 DNA疫苗所诱生的VNTR特异的CTL应答和抗体应答显著增强.     

卵泡刺激素受体主动免疫对雄性小鼠生育力影响的初步研究

探索卵泡刺激素受体 (FSHR)作为男性免疫避孕疫苗的可能性。利用分子生物学方法克隆表达卵泡刺激素受体 -谷胱甘肽转硫酶 (FSHR- GST)融合蛋白 ,以此作为免疫原免疫雄性小鼠 ,观察免疫应答反应及其对生育力的影响。结果表明 ,在大肠杆菌中表达出 FSHR- GST融合蛋白 ,免疫家兔及小鼠后均产生免疫应答 ,免疫后的雄性小鼠仍具生育能力 ,免疫小鼠睾丸精曲小管出现生精细胞减少现象。提示 FSHR主动免疫对雄性小鼠生育力无影响     

Animal model of isolated gonadotropin deficiency. I. Hormonal responses to LHRH immunoneutralization

Five intact male mongrel dogs, characterized by an episodic secretory pattern of LH and normal serum testosterone concentrations, were actively immunized against LHRH by subcutaneous injections of 200 micrograms of a LHRH-human serum albumin conjugate at 0, 4, and 8 weeks. After 12 weeks, two dogs having the highest antibody titers to LHRH (25% and 51% binding of 125I-LHRH in serum diluted 1:1000 B/Bo) had low to nondetectable serum concentrations of LH and testosterone, whereas serum FSH concentrations were significantly reduced in only one of these dogs. Immunocytochemical techniques showed that the pituitaries of these same two dogs had smaller and fewer LH immunoreactive gonadotropes than did the pituitaries of another three immunized-nonaffected dogs or of the five nonimmunized control dogs. The two LHRH-immunized dogs characterized by hypogonadotropism also had reduced testis (4.0 and 4.0 g) and prostate (2.1 and 1.7 g) weights when compared to control dogs (testis: 12.1 +/- 1.0 g and prostate: 9.2 +/- 1.9 g). LHRH antibody titers in three immunized dogs were demonstrable (8.1, 9.8, and 14.2% B/Bo), but effects on LH, FSH, and testosterone concentrations, pituitary gonadotropes, and reproductive tissue weights were not apparent. The similarity in hormonal and tissue responses observed between dogs effectively immunized against LHRH and men with isolated gonadotropin deficiency suggests that the LHRH-immunized dog may provide a suitable experimental model for the study of patients with isolated gonadotropin deficiency.     

Semisynthetic anti-LHRH vaccine causing atrophy of the prostate

A modified luteinizing hormone releasing hormone (LHRH) analog (D-lys)6 was synthesized and linked to epsilon-amino caproic acid at the 6th position. The free amino group thus generated was used for conjugation to diphtheria toxoid. Immunogenicity and bioefficacy studies of this conjugate vaccine were carried out in rodents. All immunized animals produced antibodies reactive with native LHRH. A gradual decline in testosterone levels accompanied the rise in anti-LHRH titers. On necropsy, after 10 weeks, a significant reduction in the relative weights of testes and accessory sex organs was noticed. Of particular significance was the marked atrophy of the prostate, indicating a possible therapeutic application of this vaccine in the treatment of androgen-dependent carcinoma of the prostate.     

Neoadjuvant flutamide monotherapy for locally confined prostate cancer

BACKGROUND: We compared the clinical effects and impact on quality of life (QOL) of patients who received a 3-month course of flutamide monotherapy before radical prostatectomy with those who received a 3-month course of luteinizing hormone-releasing hormone (LHRH) agonist monotherapy. METHODS: Thirty-seven patients with non-metastatic prostate cancer were enrolled in this study (19, flutamide; 18, LHRH agonist). The rates of change of serum prostate-specific antigen (PSA) and testosterone levels, downsizing of prostate volume, the rate of organ confined disease, adverse effects and perioperative scores measured using the European Organization for Research and Treatment of Cancer Prostate Cancer Quality of Life Questionnaire (EORTC-P) and the Sapporo Medical University Sexual Function Questionnaire (SMUF) were analyzed. RESULTS: At radical prostatectomy, pathological variables were not significantly different in the two groups. Serum testosterone level was significantly higher (mean 359.2 compared to 10.5, P < 0.001), complete response rate of PSA (13% compared to 57%, P = 0.028) and rate of downsizing of prostate volume (mean, -17.7% compared to -35.4%, P = 0.038) were significantly lower in the flutamide group than in the LHRH group. After neoadjuvant hormone therapy, the scores on the sexual problem domain of EORTC-P (P = 0.033) and sexual desire score of SMUF (P = 0.021) were significantly higher in the flutamide group than in the LHRH group. At a median follow-up of 34 months after prostatectomy, biochemical failure-free survival rate in the flutamide group did not differ from that in the LHRH group. CONCLUSION: This study suggests that flutamide monotherapy can be an acceptable modality as an option for neoadjuvant hormone therapy.     

Human chorionic gonadotrophin (hCG) stimulates spermatogenesis in immature mice in vivo

Background/Purpose: Spermatogenesis in postnatal testes is controlled by the hypothalamic-pituitary-gonadal axis. To determine if pituitary hormones can induce precocious spermatogenesis once primary spermatocytes (PS) have formed, prepubertal mice were treated with human chorionic gonadotrophin (hCG). Methods: Day 12 immature mice (n = 10) were injected every third day with hCG (3 or 6 IU) dissolved in 100 [mu ]L phosphate-buffered saline (PBS). Control mice (n = 10) were either uninjected or injected with 100 [mu ]L PBS alone. On day 20 to 22 the excised testes were examined histologically with tubule counts. Results: HCG-treated mice had fewer tubules at stage I (P [lt ] .001) and more at stage III than the PBS-treated group (P [lt ] .001). Mean thickness of the round spermatid layer per tubular cross section in the hCG-treated group was significantly increased compared with the PBS-treated group (P [lt ] .01). Similarly, the percentage of the tubules at stage III (containing round spermatids) in the hCG-treated group was significantly increased, from 25% to 71%, compared with the PBS-treated group (P [lt ] .01). With increasing doses of hCG the testosterone levels were significantly higher than in controls (P [lt ] .01), but hCG did not alter testis weight or position. Conclusions: These results show that hCG stimulates the transformation of PS to round spermatids even in immature mouse testes. These findings suggest that hCG treatment of prepubertal cryptorchid boys may initiate premature spermatogenesis.     

Erectile function and nocturnal penile tumescence in patients with prostate cancer undergoing luteinizing hormone-releasing hormone agonist therapy.

BACKGROUND: Luteinizing hormone-releasing hormone (LHRH) agonists have been widely used as effective agents in endocrine therapy for prostate cancer. Continuous administration of the drug results in profound suppression of testicular androgen production. However, the side effects on erectile function have not been fully investigated. METHODS: We studied the influences of testosterone suppression on male sexual function and nocturnal penile tumescence in nine sexually active patients with prostate cancer who were treated with an LHRH agonist. RESULTS: Following reduction of serum testosterone concentrations to a castration level by the administration of the LHRH agonist, sexual desire, sexual interest and sexual intercourse were totally annulled, with significant changes in frequency, magnitude, duration and rigidity of nocturnal erections observed in all patients. CONCLUSIONS: These results demonstrate that the LHRH agonist strongly suppresses erectile function and sexual activity. Taking into account the quality of sexual function for relatively young and sexually active patients and their partners, it is necessary to establish effective modalities that minimize the adverse effects on sexual function for the treatment of patients with prostate cancer.     

睾酮治疗对右心衰竭雄性大鼠心功能影响的研究

目的:临床实践及动物模型中发现雄性心力衰竭个体的性激素水平降低,尤其血浆睾酮水平降低。本研究拟在构建右心衰竭大鼠模型的基础上,采用睾酮生理补充疗法改善心功能,并探讨雄激素心脏保护作用的可能机制。方法:43只SD健康雄鼠随机分为3组:右心衰组(RHF,n=15):野百合碱一次性腹腔注射6周构建右心衰竭大鼠模型;睾酮治疗组(TT,n=15):大鼠野百合碱注射后第3天开始睾酮生理性补充;对照组(CON,n=13):同期等量生理盐水替代,其余处理同前。每2周各组大鼠取外周血,分别化学发光法、酶联免疫吸附法测外周血浆睾酮、TNF-α水平,流式细胞术检测外周血CD34+细胞含量。于第6周各组存活大鼠行右心系统血流动力学测定,取心脏、肺脏、肝脏组织行常规病理及免疫组化检测。结果:RHF组各项指标显示大鼠右心衰竭模型造模成功,且血浆睾酮水平逐渐下降、TNF-α水平增加明显;TT组大鼠睾酮生理性补充治疗后心功能改善明显,但不能完全逆转,血浆睾酮水平较RHF组大鼠有所恢复,TNF-α减少有显著性差异;TT组大鼠、RHF组大鼠外周血CD34+细胞含量、免疫组化示右心室心肌细胞CD34+表达无明显差异。结论:睾酮生理补充能够改善右心衰竭雄性大鼠的心功能,可能通过抑制炎症因子TNF-α等的释放,而非通过自体动员骨髓干细胞的途径起作用。睾酮生理补充可以作为右心衰竭治疗的新辅助手段之一。     

壳聚糖作为基因递送载体的MUC1基因疫苗治疗胰腺癌的实验研究

【摘要】 目的 研究以壳聚糖作为基因递送载体的MUC1基因疫苗诱导小鼠产生特异性体液及细胞免疫应答的作用,为壳聚糖联合MUC1基因疫苗用于胰腺癌的免疫治疗提供初步实验依据。方法 将30只小鼠平均分3组,分别接种壳聚糖-MUC1质粒、MUC1质粒及空质粒,通过ELISA法检测小鼠血清MUC1抗体生成情况,通过LDH释放法测定CTL对Capan-2细胞的杀伤活性。结果 接种壳聚糖-MUC1质粒后,小鼠血清抗MUC1抗体水平及CTL对Capan-2细胞的杀伤率均明显升高,与其它2组相比有显著性差异(P<0.05)。 结论 壳聚糖作为一种基因递送载体的MUC1基因疫苗能够有效诱导小鼠产生抗MUC1特异性抗体及产生杀伤MUC1+细胞的CTL,为壳聚糖联合MUC1基因疫苗用于胰腺癌的免疫治疗提供了初步的实验基础。     

Effect of surgical trauma on epithelial cell adhesion molecule (GA-733) vaccine–induced tumor resistance

BACKGROUND: Surgical trauma inhibits immune function. Our goal was to study the effect of surgical intervention on the development of the immune response to epithelial cell adhesion molecule (EpCAM [GA-733]), a tumor-associated protein used for vaccination in colon cancer. METHODS: Recombinant GA-733 and monophosphoryl-lipid A (MPLA) were incorporated into biodegradable beads and implanted in the following groups of mice: control, insufflation, and laparotomy. After surgery, the mice were inoculated with GA-733-transfected C26 cells (C26-EpCAM). Plasma anti-GA733 IgG antibodies were detected in enzyme-linked immunoassay (ELISA). Killing specific to GA-733 was assayed by C26-EpCAM-killing assay. RESULTS: The difference in tumor size between immunized and nonimmunized animals was statistically significant only in control mice (p < 0.05). Greater cytotoxic response to C26-GA733 developed in all immunized mice groups than in their respective controls. However, anti-GA733 IgG increased significantly in the control and insufflation groups, but not in the laparotomy group. CONCLUSIONS: Combined GA-733 vaccine allows reduction of tumor growth in control but not in surgically managed animals. This vaccine can induce a specific-cell and antibody-mediated immune response. Open surgery leads to a decreased antibody response to the GA-733 tumor vaccine.