Vaccination of Japanese patients with advanced melanoma with peptide, tumor lysate or both peptide and tumor lysate-pulsed mature, monocyte-derived dendritic cells

We performed a clinical trial to assess the feasibility and efficacy of immunotherapy with peptides, tumor lysate or both peptides and tumor lysate-pulsed mature, monocyte-derived dendritic cells (DC) for advanced malignant melanoma patients that are resistant to conventional therapies. Sixteen patients were enrolled in this trial. All patients received DC vaccines i.d. in the proximal thigh, close to the inguinal lymph nodes, one treatment per week or 2 weeks. Several factors such as clinical findings, computed tomography (CT) images, delayed type hypersensitivity (DTH) response, enzyme-linked immunosorbent spot (ELISPOT) assay, and immunohistochemistry in primary, metastatic lesions and the DTH site were evaluated. Clinical results through DC vaccination were as follows: in 11 evaluable cases, three stable disease, six progression of disease and two disease-free from the time of study entry to the completion of one vaccination course. One patient showed reduction of the tumors in the metastases on chest CT during the first and second course of DC vaccination. Ten out of 14 evaluable cases showed positive DTH responses to more than one treatment with melanoma peptides or tumor lysate. Eight out of 13 evaluable cases showed positive immunological responses to more than one treatment with melanoma peptides or tumor lysate in an ELISPOT assay. As for the experiences with toxicity and adverse reactions, autosensitization dermatitis-like eruptions appeared in five cases during DC vaccination. No severe adverse effects were seen in any of the patients. In our study, the clinical efficacy in prolongation of the patients' survival was confirmed. At the same time, cancer immunoediting of the tumor was also found. It will be necessary to improve the tumor-specificity of this therapeutic approach and to analyze the mechanism(s) of tumor escape from immunosurveillance in melanoma.     

注射用头孢哌酮钠他唑巴坦钠中细菌内毒素的检测

目的探讨注射用头孢哌酮钠他唑巴坦钠细菌内毒素的检测方法。方法按照中国药典2000版二部附录细菌内毒素检查法要求，以干扰试验测定。结果本药细菌内毒素限值为0．15EU／ml，在3．333-0．400mg／ml的浓度范围内，不干扰细菌内毒素试验。结论细菌类毒素凝胶法较家兔热原法具有灵敏度高，快速简便易行厦重现性好等优点，本品可用细菌内毒素检查凝胶法代替家兔热原法。     

卡介苗热休克蛋白70对肿瘤细胞裂解物免疫原性的影响

的：观察卡介苗热休克蛋白70（BCGHSP70）对肿瘤细胞裂解物（TCL）免疫原性的影响,为TCL相关肿瘤疫苗的制备提供依据。方法：应用冻融法制备B16黑色素瘤细胞裂解物B16TCL,将BCGHSP70与B16TCL在体外混合制备BCGHSP70-B16TCL。将C57BL/6小鼠随机分为BCGHSP70-B16TCL组、PBS组、BCGHSP70组和B16TCL组,分别于第1和14天皮下免疫BCGHSP70-B16TCL、PBS、BCGHSP70和B16TCL。于末次免疫后第7天处死小鼠,MTT法观察不同免疫组小鼠脾淋巴细胞在B16TCL体外刺激后的增殖情况,计算刺激指数（SI）。结果：应用冻融法能够使B16细胞完全裂解,所制备的B16TCL中含有丰富的蛋白成分。与PBS组、BCGHSP70组和B16TCL组比较,BCGHSP70-B16TCL组免疫小鼠脾脏明显增大;BCGHSP70-B16TCL免疫组SI高于PBS免疫组（P=0.00）、 B16TCL免疫组（P=0.01）和BCGHSP70免疫组（P=0.00）。结论：BCGHSP70　能够增强B16TCL的免疫原性,提示BCGHSP70可能作为有效的免疫佐剂用于TCL相关肿瘤疫苗的研究。     

乳鼠心肌裂解液对S180小鼠移植瘤抑瘤作用的实验研究

目的探讨乳鼠心肌裂解液对S180小鼠移植瘤生长的影响。方法提取制备乳鼠心肌细胞裂解液（car-diac muscle cell lysate of neonaterat,CMCLnr）,建立S180小鼠移植瘤模型,以顺铂组为阳性对照组,生理盐水为阴性对照组,CMCLnr再分别分为30mg/kg、60mg/kg、90mg/kg三个不同的剂量组腹腔给药,然后观察不同实验组对S180小鼠移植瘤生长的影响。结果各组小鼠移植瘤接种成活率均为100%。CMCLnr各剂量组肿瘤出现时间均晚于阴性对照组（P〈0.05）,实验各组肿瘤瘤重均小于阴性对照组,肿瘤抑制率DDP、CMCLnr（30mg/kg、60mg/kg、90mg/kg）分别为81.23%、63.09%、65.44%、66.06%,与阴性对照组比较。差异有统计学意义（P〈0.001）,且无明显剂量依赖性;CMCLnr各组的小鼠体重增加均大于生理盐水组,而顺铂组明显抑制小鼠生长（P〈0.05）。结论乳鼠心肌裂解液对S180小鼠移植瘤生长具有一定的抑制作用,且对小鼠无不良反应。     

盐酸普罗帕酮注射液细菌内毒素检查方法的确立

目的：验证盐酸普罗帕酮注射液细菌内毒素检查法的可行性。方法：通过抑制／增强试验确定盐酸普罗帕酮注射液的有效稀释浓度和细菌内毒素限值。结果：0．35mg/ml的盐酸普罗帕酮溶液无干扰作用，其内毒素限值符合规定。结论：盐酸普罗帕酮注射液可用细菌内毒素检查法来控制其细菌内毒素含量。     

Polymyxin B use does not ensure endotoxin-free solution

Polymyxin B is often added to in vitro samples to 'ensure' that endotoxin activity is removed. We present data, from the standard rabbit pyrogen test and the Limulus amebocyte lysate assay, that polymyxin B bound to a gel support will bind some, but not all, endotoxin. These data, in conjunction with previously published data by Morrison and Curry (1979), indicate that those studies that have relied on polymyxin B to inactivate endotoxin must be re-evaluated.     

Effects of calcium and limited proteolysis on membrane-bound and releasable dopamine β-hydroxylase in adrenomedullary catecholamine granules

HELLE, K.B., PIHL, K.E. & SERCK-HANSSEN, G. 1985. Effects of calcium and limited proteolysis of membrane-bound and releasable dopamine β-hydroxylase in adrenomedullary catecholamine granules. Acta Physiol Scand 125 , 423–427. Received 9 January, accepted 6 May 1985. ISSN 0001–6772. Department of Physiology, University of Bergen, Norway. Bovine chromaffin granules were shown to contain two potent proteolytic systems resulting in limited proteolysis of granule proteins at pH 6.0 in the cold in the presence of inhibitors of serine and thiol proteases. Calcium, whether added during lysis or remaining bound to the lysate protein during dialysis in non-chelating solutions, enhanced recoveries of total immunoreactive dopamine β-hydroxylase (14% of total protein) and soluble enzyme (9% of lysate protein) due to degradation of chromogranins. A pepstatin A-blockable, catepsin D-like proteolytic system converting membrane-bound enzyme to its soluble counterpart at pH 6.0 was detected in the granule membrane fraction.     

自身骨髓瘤抗原致敏树突细胞介导特异性CTL反应的研究

目的：研究多发性骨髓瘤(MM)患者肿瘤冻融物致敏的树突细胞(DC)能否诱导特异性细胞毒T淋巴细胞(CTL)反应。方法：用MM患者骨髓CD34^ 细胞诱生DC。将MM患者骨髓瘤冻融物冲击致敏DC。MTT法检测骨髓瘤抗原致敏及非致敏DC诱导的自身T细胞对不同靶细胞(患者骨髓瘤细胞、K562细胞)的杀伤率。结果：骨髓瘤冻融物致敏DC诱导的自身T细胞对患者骨髓瘤细胞的杀伤远大于对K562细胞的杀伤。结论：患者骨髓瘤冻融物致敏的DC能有效诱导自身T细胞特异性抗瘤免疫。