治疗用卡介苗的研制

高浓度卡介苗临床用于膀胱癌术后化疗已有数十年。经临床使用其抑制浅表型膀胱癌术后复发效果显著。为满足临床患者的使用,需研制一种适合的治疗用卡介苗制剂。本文报道了三批治疗用卡介苗的研制过程及本所质管处,中检所的检定结果。样品现已通过检定,正申请试生产文号     

膀胱癌术后卡介苗灌注的护理配合

膀胱癌是泌尿系统常见的肿瘤,确诊后应首选手术治疗.据资料记载,手术切除肿瘤或膀胱部分切除术后,约有2/3患者肿瘤复发,故一般术后采用膀胱内药物灌注治疗以防复发[1].我院自1995年以来对32例膀胱癌患者术后采用卡介苗(BCG)灌注治疗,疗效好,副作用少,现就护理体会总结如下.     

皮下预免疫卡介苗提高卡介苗膀胱灌注诱导的外周特异性免疫应答的实验研究

膀胱内灌注卡介苗(bacillus Calmette-Guerin, BCG)是经尿道膀胱肿瘤切除术(transurethral resection of bladder tumor, TURBt)后的中高危非肌层浸润性膀胱癌(non-muscle-invasive bladder cancer,NMIBC)患者的标准辅助治疗。然而,其作用机制仍不完全清楚。本研究利用小鼠皮下免疫模型和膀胱内灌注模型,使用多色流式染色技术检测膀胱内的免疫细胞浸润情况及外周纯蛋白衍生物(purified proteinderivate,PPD)特异性细胞免疫水平,探究皮下预免疫BCG对膀胱内BCG灌注诱导的外周特异性免疫应答的影响。研究结果显示,膀胱内BCG灌注3次后,小鼠脾脏中PPD特异性IFN-γ⁺CD4⁺T细胞(P<0.001)和CD8⁺T细胞(P<0.05)的比例显著上升,并维持在一定的水平;同时,与单纯膀胱内BCG灌注组相比,BCG预免疫后再接受膀胱内BCG灌注的小鼠膀胱局部浸润的CD4⁺T、NK...     

国内外卡介苗治疗膀胱癌的经验与展望

1976年,美国首次报道,用卡介苗经膀胱灌注,治疗膀胱癌有良好疗效;次年,我国亦作了同样的介绍。实验证明,卡介苗可增强和提高患者的局部和全身免疫,术后治疗的复发率;国外资料为32.91%（化疗组40.61%,未治疗组为87.67%）;我国资料为17.32%（化疗组为34.01%,未治疗组为60.90%）,副反应主要为尿痛、尿频、轻度血尿、低热等一过性反应;很少见全身性副反应。国内所见副反应率低于国外资料。目前,用于治疗膀胱癌的卡介苗,已获本国批准的有5个以上制品;我国同类产品已完成临床Ⅱ期试验,正在申报审核中。本品治疗膀胱癌是一种安全、效好的首选药品。     

卡介苗（BCG）膀胱冲洗预防膀胱癌复发的研究

探讨不同方法预防膀胱肿瘤术后复发的效果.将1994～2001年85例膀胱肿瘤术后患者分为2组,BCG(卡介苗)80 mg加生理盐水50 mL、噻替哌50 mg加生理盐水50 mL,行膀胱内冲洗预防膀胱肿瘤复发,评价其疗效.随访2～5 a,BCG组复发率37.1%,噻替哌组复发率50.8%.2组比较有统计学差异.结果显示,BCG和噻替哌均能预防膀胱肿瘤术后复发,但BCG优于噻替哌.     

卡介苗菌基因组DNA对小鼠腹腔巨噬细胞激活效应的初步研究

为了比较研究卡介苗菌基因组DNA和卡介苗菌激活小鼠腹腔巨噬细胞的抗结核免疫应答过程及免疫作用,用卡介苗菌基因组DNA、卡介苗菌和生理盐水分别皮内注射小鼠第30、60 d后,分别采用Griess法、化学法、ELISA法检测小鼠腹腔巨噬细胞产生的NO、H2O2以及IL-12、TNF-α的表达.研究结果,卡介苗菌基因组DNA和卡介苗菌皮内接种小鼠后能显著诱导小鼠腹腔巨噬细胞分泌表达IL-12和TNF-α.卡介苗菌基因组DNA组和卡介苗菌组相比较,无明显差异,与生理盐水组比较,其差异均有统计学意义(P<0,05);卡介苗菌基因组DNA和卡介苗菌皮内接种小鼠后也能诱导小鼠腹腔巨噬细胞产生NO、H2O2,卡介苗菌基因组DNA组和卡介苗菌组相比较,无明显差异,与未免疫组比较均具有统计学意义(P<0.05).研究结果表明,卡介苗菌基因组DNA皮内接种小鼠后能诱导小鼠腹腔巨噬细胞活化并产生较强的激活效应,且该效应于卡介苗菌无明显差异.     

卡介苗溶菌黏膜免疫调理剂对激活荷瘤小鼠免疫功能的研究

观察卡介苗溶菌黏膜免疫调理剂对荷瘤小鼠免疫功能的激活。将SPF小鼠分组后用黑色素瘤B16和肝癌实体瘤H22感染,随后以灌胃途径引入本调理剂,分别发现它能促进荷瘤小鼠血清溶菌酶含量上升和T细胞介导的迟发型变态反应增强。试验结果表明,卡介苗溶菌黏膜免疫调理剂对荷瘤小鼠有升高其血清溶菌酶含量和增强迟发型变态反应的作用。     

卡介苗作用大鼠膀胱肿瘤细胞／正常移行上皮细胞后TNF—α．、IL-10、IFN-γ的检测及意义

目的：观察卡介苗（BCG）单独作用膀胱肿瘤细胞、正常膀胱移行上皮细胞及其代谢产物作用上述细胞后细胞生长情况及各自细胞培养液上清液中细胞因子（TNF-α．、IL-10、IFN-γ）浓度的变化,探讨其在卡介苗治疗膀胱肿瘤中可能的作用机制。方法：构建大鼠膀胱肿瘤模型,并原代培养大鼠膀胱肿瘤细胞及正常膀胱移行上皮细胞。分别用BCG,普通培养液和细胞培养的代谢产物作用上述细胞。酶联接免疫吸附剂测定法（ELISA法）检测各组细胞上清液中肿瘤坏死因子-α（TNF-α）、白细胞介素-10（IL-10）、干扰素-γ（IFN-γ）的浓度。结果：ELISA法检测各组细胞上清液中TNF-α、IL-10的浓度改变有显著差异,而IFN-γ的浓度无显著差异。结论：BCG可以直接刺激肿瘤细胞自身分泌细胞因子（TNF-α、IL-10）参与调节抑制肿瘤细胞的生长。     

TUR-BT术后膀胱灌注A群链球菌联合丝裂霉素治疗高危非肌层浸润性膀胱癌的疗效观察

目的：总结经尿道膀胱肿瘤电切（TUR-BT）术后辅以沙培林（注射用A群链球菌）联合丝裂霉素（MMC）膀胱内灌注治疗高危膀胱癌的疗效。方法：回顾性研究2009年1月~2012年8月我院收治的符合高危非肌层浸润性膀胱癌患64例,观察组：TUR-BT术后行沙培林联合MMC膀胱内灌注化疗32例,对照组;MMC单药灌注32例。结果：观察组患者,平均年龄63.7岁,治疗随访时间为6~54个月,中位时间27.3个月。治疗随访期间有3例患者出现膀胱内肿瘤复发（9.3%）,1例患者疾病进展,发展为肌层浸润性膀胱癌,于术后7个月死亡（3.1%）。与对照组患者相比,疾病复发率及进展率均明显改善。结论：高危非肌层浸润膀胱癌临床复发率、进展率高,TUR-BT术后沙培林联合MMC膀胱内灌注通过局部化疗及免疫治疗联合,可有效控制疾病的复发和进展,降低患者接受膀胱部分切除或膀胱全切手术的机率,值得推广。     

阴道用乳杆菌活菌胶囊对改善子宫全切术后性生活质量的效果分析

目的 探讨阴道用乳杆菌活菌胶囊改善子宫全切术后女性性生活质量的效果。方法 选择2013年1月至2014年12月,广西壮族自治区人民医院妇科因子宫良性病变行子宫全切术的患者共60例,随机分为研究组（阴道用乳杆菌活菌胶囊治疗组）及对照组（安慰剂组）各30例,术后第6个月(即治疗前)及术后第7个月（即治疗后）所有患者均检测阴道pH值及完成PISQ-12问卷调查。结果 研究组患者治疗1个疗程后阴道pH值较治疗前明显下降,PISQ-12问卷评分较治疗前升高,对照组患者治疗前后阴道pH值和PISQ-12问卷评分无明显变化,两组间阴道pH值和PISQ-12问卷评分的差异有统计学意义（P＜0.05）。结论 阴道用乳杆菌活菌胶囊可有效改善子宫全切术后女性性生活质量。     

A possible mechanism of intravesical BCG therapy for human bladder carcinoma: involvement of innate effector cells for the inhibition of tumor growth

Intravesical bacillus Calmette-Guerin (BCG) therapy is considered the most successful immunotherapy against solid tumors of human bladder carcinoma. To determine the actual effector cells activated by intravesical BCG therapy to inhibit the growth of bladder carcinoma, T24 human bladder tumor cells, expressing very low levels of class I MHC, were co-cultured with allogeneic peripheral blood mononuclear cells (PBMCs) with live BCG. The proliferation of T24 cells was markedly inhibited when BCG-infected dendritic cells (DCs) were added to the culture although the addition of either BCG or uninfected DCs alone did not result in any inhibition. The inhibitory effect was much stronger when the DCs were infected with live BCG rather than with heat-inactivated BCG. The live BCG-infected DCs secreted TNF-α and IL-12 within a day and this secretion continued for at least a week, while the heat-inactivated BCG-infected DCs secreted no IL-12 and little TNF-α. Such secretion of cytokines may activate innate alert cells, and indeed NKT cells expressing IL-12 receptors apparently proliferated and were activated to produce cytocidal perforin among the PBMCs when live BCG-infected DCs were externally added. Moreover, depletion of γδ T-cells from PBMCs significantly reduced the cytotoxic effect on T24 cells, while depletion of CD8β cells did not affect T24 cell growth. Furthermore, the innate effectors seem to recognize MICA/MICB molecules on T24 via NKG2D receptors. These findings suggest the involvement of innate alert cells activated by the live BCG-infected DCs to inhibit the growth of bladder carcinoma and provide a possible mechanism of intravesical BCG therapy.     

The ‘experimental stable’ of the BCG vaccine: safety, efficacy, proof, and standards, 1921–1933

The anti-tuberculosis BCG (Bacille Calmette-Guérin) vaccine was conceived and developed between 1905 and 1921 at Pasteur Institutes in France. Between 1921 and A. Calmette’s death in 1933, the vaccine went through a first period of national and international production and distribution for its use in humans. In France these activities were exclusively carried out by Calmette and his collaborators at the Pasteur Institute in Paris. Initially improvised production in a small room in the cellar gave way in 1931 to the construction of the spacious and magnificent ‘New laboratories for research on tuberculosis and the preparation of the BCG’ within the premises of the Pasteur Institute. Presentation and image-building of the vaccine in France insisted on the fact that the BCG was not a commercial specialty but distributed free of charge. The technical monopoly of its production nevertheless lay with the Paris Pasteur Institute and standardization of scientific proof of safety, efficacy and stability was dominated by that Institute in France. In contrast, the international production and distribution of the vaccine was entrusted and transferred, free of charge, to trustworthy laboratories outside France. Multiplication of producers and users led to an increased need for standardization. For this process the analysis distinguishes between the standardization of scientific proof concerning safety, efficacy and stability of the vaccine and standardization of its medical uses. Whereas standardization was rather successful in the inter-war period in France, the international efforts remained rather unsuccessful. Only after world war II under Scandinavian leadership and in the context of mass vaccination programs supported by the WHO and UNICEF was the international standardization effectively implemented and succeeded at least to some extend.     

BCG Vaccination Protects against Experimental Viral Infection in Humans through the Induction of Cytokines Associated with Trained Immunity

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The immune enhancement of propolis adjuvant on inactivated porcine parvovirus vaccine in guinea pig

Two experiments were carried out. In immune response test, the immune enhancement of propolis, oilemulsion and aluminium salt were compared in guinea pig vaccinated with inactivated porcine parvovirus (PPV) vaccine. The result showed that three adjuvants could enhance antibody titer, T lymphocyte proliferation, IL-2 and IL-4 secretion of splenic lymphocyte. The action of propolis was similar to that of oilemulsion and superior to that of aluminium salt, especially in early period of vaccination propolis could accelerate antibody production. In immune protection test, the effects of three adjuvants on PPV infection were compared in guinea pig vaccinated with PPV vaccine then challenged with PPV. The result showed that propolis and oilemulsion could enhance the antibody titer, IL-2 and IL-4 content in serum and decrease the PPV content in blood and viscera. In the effect of improving cellular immune response, the propolis was the best. These results indicated that propolis possessed better immune enhancement and would be exploited into a effective adjuvant of inactivated vaccine.     

Phase I study of a MUC1 vaccine composed of different doses of MUC1 peptide with SB-AS2 adjuvant in resected and locally advanced pancreatic cancer

MUC1 is a glycoprotein overexpressed in tumors as a hypoglycosylated form. A vaccine composed of a 100–amino acid peptide corresponding to five 20–amino acid long repeats, and SB-AS2 adjuvant, was tested in a phase I study for safety, toxicity, and ability to elicit or boost MUC1-specific immune responses. Patients with resected or locally advanced pancreatic cancer without prior chemotherapy or radiotherapy were eligible. Escalating doses of the peptide (100, 300, 1,000, and 3,000 g) were admixed with SB-AS2 and administered intramuscularly every 3 weeks for three doses, in cohorts of four patients. Sixteen patients were enrolled. Common adverse effects were grade 1 flu-like symptoms, tenderness, and erythema at the injection site. Delayed-type hypersensitivity (DTH) sites showed few or no T cells prevaccination (Pre V), but increased T-cell infiltration postvaccination (Post V). There was an increase in the percentage of CD8⁺ T cells in the peripheral blood Post V. An increase in total MUC1-specific antibody was seen in some patients, and several patients developed IgG antibody. Two of 15 resected pancreatic cancer patients are alive and disease free at follow-up of 32 and 61 months. MUC1 100mer peptide with SB-AS2 adjuvant is a safe vaccine that induces low but detectable mucin-specific humoral and T-cell responses in some patients. No difference was seen between different peptide doses. Further evaluation is warranted to examine the effect on disease-free survival and overall survival, especially in early disease and in the absence of immunosuppressive standard therapy.Work presented in part at the 36th Annual American Society of Clinical Oncology Meeting, New Orleans, LA, May 2000