Dose and Load Studies for Subcutaneous and Oral Delivery of Poly(lactide-co-glycolide) Microspheres Containing Ovalbumin

Poly(lactide-co-glycolide) microspheres containing different loads of OVA (0.05, 0.1, 0.5 and 1.0% w/w) were manufactured by a w/o/w emulsion/solvent evaporation method. Low load efficiencies of less than 20% were observed. Normal size distributions with mean volume diameters ranging from 3.7 to 4.7 µm were obtained for different batches. The in vitro release of OVA from different loaded microspheres showed an expected burst release with all batches. The in vivo dose study (1, 10, 25, 50 µg of OVA) was performed by subcutaneous and oral inoculation in mice by single (0 week) or double (0 and 3 weeks) administration of PLGA 50/50 microspheres containing 0.1% OVA. Subcutaneous administration showed an immune response (serum Ig levels by ELISA) statistically (Fishers paired t-test; P < 0.05) above OVA saline negative controls at 3, 6 and 12 weeks after administration. Oral administration of microspheres produced statistically higher systemic immune responses at the higher doses. Single and double inoculation orally and subcutaneously produced similar serum antibody levels. The in vivo load study was performed by subcutaneous and oral administration to mice of 25 µg OVA contained in various loaded (0.05, 0.1, 0.5 and 1.0% w/w) microspheres. Serum immune responses at 3, 6, and 12 weeks after inoculation were statistically above OVA saline controls and were inversely proportional to the OVA load using either route. This observation suggested a relationship between the number of microspheres delivered and the in vivo serum response. Single subcutaneous administration of 0.05 or 0.1% OVA loaded PLGA 50/50 microspheres induced larger immune responses compared with complete Freunds adjuvant.     

用PCR体外定点突变技术诱导霍乱毒素A亚基突变体的构建

目的： 有效去除霍乱毒素Ａ 亚基（ ＣＴ Ａ） 的毒性作用而保存其佐剂性能。方法： 采用ＰＣＲ 体外定点突变技术（ＰＣＲ ＳＤＭ） ,设计两对引物,引入一个突变位点,通过重叠延伸法两次ＰＣＲ 扩增,使ＣＴ Ａ 编码基因第６３ 位密码子由ＣＴＣ 突变为ＴＴＴ,亦将扩增片断克隆入ＰＵＣ１９ 载体。结果： ＤＮＡ 测序结果表明在预期位点已发生突变,用ＰＣＲ 诱导成功ＣＴ Ａ 突变体。结论： ＰＣＲ 诱导突变准确、简便,为深入研究ＣＴ 免疫佐剂的作用打下了基础     

麦粒灸对佐剂性关节炎大鼠细胞免疫功能的影响

目的　探讨麦粒灸对佐剂性关节炎 ( A A)大鼠细胞免疫功能的影响。方法　以佐剂性关节炎作为类风湿关节炎大鼠模型 ,分为正常对照组、模型组、麦粒灸治疗组、麦粒灸加去肾上腺组、麦粒灸加假手术组。麦粒灸“肾俞”、“足三里”,检测IL-1、IL-2、IL-6含量变化。结果　模型组大鼠腹腔巨噬细胞 IL-1、IL-6的活性比正常大鼠明显升高 ( P<0 .0 1)。麦粒灸组大鼠 IL-1、IL-6活性较模型组比明显下降 ( P<0 .0 5 )。麦粒灸加去肾上腺组 IL-1、IL-6活性未见明显变化 ( P>0 .0 5 ) ,假手术组IL-1活性极显著降低 ( P<0 .0 1) ,IL-6比模型组明显为低 ( P<0 .0 5 )。模型组大鼠脾脏 IL-2活性降低 ( P<0 .0 5 ) ,麦粒灸治疗后可使 IL-2活性极显著升高 ( P<0 .0 1) ,麦粒灸加去肾上腺组 IL-2活性与麦粒灸组及假手术组比 ( P<0 .0 5 ) ,但与模型组比无显著性差异 ( P>0 .0 5 )。假手术组 IL-2活性亦明显提高 ( P<0 .0 1) ,而与正常组无差异。结论　麦粒灸可抑制 AA大鼠 IL-1、IL -6活性。摘除肾上腺后则麦粒灸不能降低 IL -1、IL -6活性 ;麦粒灸可明显提高 IL -2活性 ,摘除肾上腺后麦粒灸则对 IL -2活性无明显影响 ;麦粒灸对 A A细胞免疫功能的调整与下丘脑 -垂体 -肾上腺的完整有关     

右酮洛芬氨丁三醇对佐剂性关节炎大鼠的治疗作用及胃肠道损伤作用的研究

目的　研究右酮洛芬氨丁三醇 (D KPT)对佐剂性关节炎大鼠的治疗作用及胃肠道损伤作用。方法　采用大鼠佐剂性关节炎模型 ,观察D KPT对佐剂性关节炎大鼠原发和继发性症状的作用以及对前列腺素E2 和胃、十二指肠的影响。结果　D KPT(2 5、5、10mg·kg-1)不仅能抑制佐剂性关节炎大鼠的原发性炎症 ,而且对继发性炎症和多发性关节炎有抑制作用。D KPT(10 -8、10 -7、10 -6 、10 -5、10 -4 mol·L-1)体外对佐剂性关节炎大鼠腹腔巨噬细胞产生过高的PGE2 有抑制作用。D KPT(10mg·kg-1)对胃、十二指肠黏膜有损伤作用 ,但损伤程度小于酮洛芬 (KP) (10mg·kg-1)。结论　D KPT对佐剂性关节炎大鼠有治疗作用 ,同时对胃、十二指肠损伤小于KP。     

万灵五香巴布膏活血通络与消肿止痛的药理作用

目的：观察万灵五香巴布膏活血通络与消肿止痛的药理作用。方法：采用小鼠光热法与扭体法镇痛试验观察止痛作用；小鼠巴豆油耳肿胀试验与大鼠佐剂关节炎抗炎试验观察消肿作用；小鼠耳廓微循环试验和大鼠急性软组织损伤试验观察活血通络作用。结果：万灵五香巴布膏能显提高小鼠光热法测得的痛阈和减少冰醋酸所致小鼠扭体反应次数；减轻巴豆油所致小鼠耳肿胀和大鼠佐剂性关节炎的肿胀度；改善小鼠耳廓微循环和减轻打击所致大鼠急性软组织损伤。结论：万灵五香巴布膏有明显的活血通络与消肿止痛药理作用。     

老年直肠癌临床特征及治疗的变迁

目的 了解近年来老年直肠癌临床特征及治疗的变迁。方法 回顾性分析10年间313例老年直肠癌患者的临床特征及治疗情况，对比第一个5年和第二个5年的临床特征及治疗方法的变迁。结果发病年龄后延，误诊率降低，早期病例增多；Dixon手术增多，辅助治疗由简单变为综合治疗方法；5年生存率明显提高，术后的复发率明显减低。结论 老年直肠癌发病年龄后延，误诊率降低，早期病例逐渐增多；治疗上更趋有效、合理。     

Peripheral blood circulating cytokeratin-19 mRNA-positive cells after the completion of adjuvant chemotherapy in patients with operable breast cancer.

BACKGROUND: The purpose of this study was to evaluate the prognostic significance of the molecular detection of cytokeratin 19 (CK-19) mRNA-positive cells in the peripheral blood of women with operable breast cancer after the completion of adjuvant chemotherapy. PATIENTS AND METHODS: Blood from 161 patients with stage I and II breast cancer, obtained after the completion of adjuvant chemotherapy, was tested by nested RT-PCR for CK-19 mRNA detection. Using univariate and multivariate analyses possible interactions with other prognostic factors and association of CK-19 mRNA detection with risk of relapse, disease-free interval (DFI) and overall survival were investigated. RESULTS: After completion of adjuvant chemotherapy, 27.3% of patients had peripheral blood CK-19 mRNA-positive cells; there was no association of this finding with any other prognostic factors or the type of chemotherapy regimen used. For patients with less than four involved axillary lymph nodes the risk of relapse was 3.81 [95% confidence interval (CI) 1.06-13.71] times higher, and the DFI was significantly reduced (P = 0.028) if CK-19 mRNA-positive cells were detectable in the blood after the completion of adjuvant chemotherapy. In contrast, for patients with four or more involved lymph nodes, the presence of CK-19 mRNA-positive cells after adjuvant chemotherapy did not significantly affect the risk of relapse or DFI. Furthermore, the risk of relapse was higher (hazards ratio 3.70; 95% CI 1.09-13.89) and the DFI was reduced (P = 0.022) for patients with detectable CK-19 mRNA-positive cells following adjuvant cyclophosphamide, methotrexate and 5-fluorouracil (CMF) as compared with epirubicin, cyclophosphamide and 5-fluorouracil (FEC) or sequential taxotere-epirubicin and cyclophosphamide (T/EC) chemotherapy. CONCLUSIONS: The detection of CK-19 mRNA-positive cells in the peripheral blood after adjuvant chemotherapy may be of clinical relevance for patients with early breast cancer and less than four involved axillary lymph nodes.     

化疗在大肠癌综合治疗中的作用

本文对化疗在大肠癌综合治疗中的作用进行评述。近年来治疗晚期与转移性大肠癌的策略有了明显的进展,新药新方法不断涌现,全身化疗能提高晚期大肠癌的缓解率,延长生存期,改善生活质量;辅助化疗已成为高危复发患者的标准治疗;新辅助化疗越来越受到重视;然而,在具体实施综合治疗中如何按照大肠癌的生物学行为,在规范化治疗的前提下,充分考虑患者潜在的预后亚群、效益-成本比、患者的依从性等因素实施个体化,选择最佳治疗方式值得研究。     

5-Fluorouracil plus leucovorin is an effective adjuvant chemotherapy in curatively resected stage III colon cancer: long-term follow-up results of the adjCCA-01 trial.

BACKGROUND: Adjuvant postoperative treatment with 5-fluorouracil (5-FU) and leucovorin in curatively resected stage III colon cancer significantly reduces the risk of cancer recurrences and improves survival. The impact of 5-FU plus leucovorin on survival and tumor recurrence was analyzed in a long-term follow-up study in comparison with the effects of 5-FU plus levamisole in the prospective multicenter trial adjCCA-01. PATIENTS AND METHODS: Patients with a curatively resected stage III (International Union Against Cancer) colon cancer were stratified according to tumor, node and grading category and randomly assigned to receive one of the two adjuvant treatment schemes: 5-FU 400 mg/m2 body surface area intravenously in the first chemotherapy course, then 450 mg/m2 x 5 days, plus leucovorin 100 mg/m2, 12 cycles (arm A), or 5-FU plus levamisole (Moertel scheme; arm B). RESULTS: Six hundred and eighty (96.9%) of 702 patients enrolled into this study were eligible. To date, 261 patients have died, 117 on arm A and 144 on arm B (P = 0.007). After a median follow-up time of 82 months, the 5-FU plus leucovorin combination significantly improved disease-free survival [79.8 months in arm A versus 69.3 months in arm B (P = 0.012)] and significantly increased median overall survival (88.9 months in arm A versus 78.6 months in arm B; P = 0.003). Adjuvant treatment with 5-FU plus levamisole as well as 5-FU plus leucovorin was generally well tolerated; only a minority of patients experienced grade 3 and 4 toxicities. CONCLUSIONS: After curative resection of a stage III colon cancer, adjuvant treatment with 5-FU plus leucovorin is generally well tolerated. This long-term follow-up study demonstrates that adjuvant treatment with 5-FU plus leucovorin given for 12 cycles is significantly more effective than 5-FU plus levamisole (Moertel scheme) in reducing tumor relapse and improving survival.     

Low incidence of secondary myelodysplasia and acute myeloid leukemia after high-dose chemotherapy as adjuvant therapy for breast cancer patients: a study by the Solid Tumors Working Party of the European Group for Blood and Marrow Transplantation.

BACKGROUND: To determine the incidence of secondary myelodysplasia (sMDS) or acute myeloid leukemia (AML) in node-positive breast cancer patients who received high-dose chemotherapy (HDCT) followed by autologous stem-cell support as adjuvant therapy. PATIENTS AND METHODS: The incidence of sMDS/AML was retrospectively assessed in 364 node-positive breast cancer patients who received HDCT followed by autologous stem-cell support as adjuvant therapy between November 1989 and December 1997 and were reported to the European Group for Blood and Marrow Transplantation registry. RESULTS: The median age of the patients was 45 years (range 22-62 years). Two hundred and ninety-one patients received peripheral blood stem cells and 55 patients received autologous bone marrow as stem-cell support. The most frequently used conditioning regimen was the STAMP-V regimen (32%), followed by melphalan-thiotepa (22%) and melphalan-mitoxantrone-cyclophosphamide (21%). The 5-year probability of overall survival is 71% (95% CI 65% to 77%). After a median follow-up of 48 months (range 1-108 months) only one case of AML was observed, resulting in a crude incidence of 0.27%. This case of AML was observed 18 months after HDCT consisting of three cycles of epirubicin and cyclophosphamide with a cumulative dose of epirubicin 960 mg and cyclophosphamide 19 g. The French-American-British type of AML was M4, and the cytogenetic analysis showed a translocation t(9;11)(p22;q23). After complete remission following high-dose cytarabine and idarubicin the patient relapsed and died. CONCLUSIONS: In contrast to patients with malignant lymphoma there seems to be no increased risk of sMDS/AML after HDCT in breast cancer. Continued monitoring is required to confirm this low incidence after a longer follow-up period.