11种天然药材及植物的抗突变与致突变同步试验报告

本文报告用ＳＯＳ噬菌体诱导试验的抗突变与致突变同步试验法，对１１种天然中药及植物甘草、枸杞、半枝莲、柴胡、丹参、黄芪、决明子、仙人球、仙人指、胡萝卜及青萝卜的抗突变及致突变性进行了筛检。经过加和不加大鼠肝微粒体酶代谢活化系统（Ｓ９）的２种试验及重复验证，结果１１种中药及植物均未发现致突变作用，甘草、枸杞、丹参、黄芪、胡萝卜、青萝卜及仙人指可抑制抗肿瘤药物丝裂霉素Ｃ（ＭＭＣ）的致突变性。     

大黄抗突变作用的研究

本文通过Ａｍｅｓ试验ＴＡ９８、ＴＡ１００菌株和ＳＯＳ反应原噬菌体诱导抗突变和致突变同步试验测试了大黄水溶性提取液的抗突变作用。结果表明大黄在４～４０ｍｇ／皿三个浓度，加或不加Ｓ９试验中对柔毛霉素、２ＡＦ所诱发的ＴＡ９８的突变、对叠氮钠、环磷酰胺所诱发的ＴＡ１００的突变有不同程度的抑制作用，并呈观剂量效应关系。对移码型致突变剂的作用较优。原噬菌体诱导抗突变和致突变同步试验发现大黄对ＭＭＣ诱发的突变也表现出明显的抑制。     

体外亚硒酸氢钠致突变和抗突变测试

亚硒酸氢钠（ＮａＨＳｅＯ３）的致突变和抗突变尚无定论，本文应用稍加修改的ＳＯＳ／Ｕｍｕ显色试验对其这方面的作用作了研究。结果发现，ＮａＨＳｅＯ３剂量在０．６７－８．００ｍｇ／ｍｌ范围内，无Ｓ９时，ＳＯＳ反应的诱导率（Ｒ）有剂量－效应了低诱导剂量为１．８６ｍｇ／ｌ；存在Ｓ９时，各剂量组的Ｒ值均接近于１。结果还揭示ＮａＨＳｅＯ３能明显抑制丝裂霉素Ｃ（ＭＭＣ）诱发的β－半乳糖苷酸（ＢＧａｓｅ）活性，其剂     

桑葚等五种可食性中药材的抗突变和致突变性初步试验研究

目的：为筛选有抗突变作用的天然可食性中药材,增加饮食中的抗癌因素,为肿瘤的人群预防提供基础依据,我们在筛选数十种抗突变天然可食性中药材的基础上,又选择了桑葚、葡萄、杏、榆钱及樱桃进行研究。方法：采用抗突变和致突变同步快速试验,试验中以00.5mg/ml的丝裂霉素C（MMC）为致突变阳性对照剂,100mg/ml的维生素C（Vit.C)为抗突变阳性对照剂,0．85％的生理盐水为既不致突变也不抗突变的阳性对照剂,并设立平行对照。做了加大鼠肝脏微粒体酶（S9）的试验和不加S9的试验,对每种检品均做重复验证试验。标准试验菌株为大肠杆菌溶源性菌GY5027,该菌遇到致突变剂即发生SOS反应产生噬菌体,对该噬菌体敏感的菌为GY4015,两菌相遇产生噬菌斑,试验前对两菌株进行了鉴定符合其原生物学特性,对受试物进行清洗沥去水滴,用无菌组织研磨器研碎,加生理盐水制成1：3液汁,加热至95℃5min自然冷却待测。菌种的活化及处理、向半固体培养基中加菌铺皿、各种培养基的配置、检品及对照剂的处理,判断结果的标准等试验程序均按抗突变和致突变同步快速试验方法中的直线加条法常规进行。处理完毕后,置于37℃恒温箱培养,6－12h内观察结果,加S9的试验在冰盘上低温快速操作,每皿中加0.5ml的S9混合液,其他操作程序与不加S9试验相同。结果：经过加和不加S9两种试验,结果该5种检品均未发现致突变毒性;桑葚、葡萄、杏和樱桃对MMC引起的致突变性有明显的拮抗作用。结论：该4种天然可食性中药材除原已知的药理作用外,有可能还具有防癌作用,值得进一步验证及研究,榆钱虽未发现抗突变作用,但也证实无诱变毒性,仍可按民间习惯食用。     

87. 桑葚等五种可食性中药材的抗突变和致突变性初步试验研究

目的：为筛选有抗突变作用的天然可食性中药材,增加饮食中的抗癌因素,为肿瘤的人群预防提供基础依据,我们在筛选数十种抗突变天然可食性中药材的基础上,又选择了桑葚、葡萄、杏、榆钱及樱桃进行研究。方法：采用抗突变和致突变同步快速试验,试验中以0.05 mg/ml的丝裂霉素C(MMC)为致突变阳性对照剂,100 mg/ml的维生素C(Vit.C）为抗突变阳性对照剂,0.85％的生理盐水为既不致突变也不抗突变的阳性对照剂,并设立平行对照。做了加大鼠肝脏微粒体酶（S9）的试验和不加S9的试验,对每种检品均做重复验证试验。标准试验菌株为大肠杆菌溶源性菌GY5027,该菌遇到致突变剂即发生SOS反应产生噬菌体,对该噬菌体敏感的菌为GY4015,两菌相遇产生噬菌斑,试验前对两菌株进行了鉴定符合其原生物学特性,对受试物进行清洗沥去水滴,用无菌组织研磨器研碎,加生理盐水制成1∶3液汁,加热至95℃ 5 min自然冷却待测。菌种的活化及处理、向半固体培养基中加菌铺皿、各种培养基的配置、检品及对照剂的处理、判断结果的标准等试验程序均按抗突变和致突变同步快速试验方法中的直线加条法常规进行。处理完毕后,置于37℃恒温箱培养,6～12 h内观察结果。加S9的试验在冰盘上低温快速操作,每皿中加0.5 ml的S9混合液,其他操作程序与不加S9试验相同。结果：经过加和不加S9两种试验,结果该5种检品均未发现致突变毒性;桑葚、葡萄、杏和樱桃对MMC引起的致突变性有明显的拮抗作用。结论：该4种天然可食性中药材除原已知的药理作用外,有可能还具有防癌作用,值得进一步验证及研究,榆钱虽未发现抗突变作用,但也证实无诱变毒性,仍可按民间习惯食用。     

香菇水提取液体外抗致突变作用

香菇水提取液体外抗致突变作用郭原健，李国华，贾继锋，傅娟玲，周宗灿（山西省防疫站０３００１２北京医科大学北京１０００８３）本文用ＳＯＳ显色法和Ａｍｅｓ试验方法观察了香菇水提取液对丝裂霉素Ｃ（ＭＭＣ）和紫外线（ＵＶ）诱导大肠杆菌ＰＱ３５ＳＯＳ反应的抑制...     

复方天然保健果茶的研制和抗突变及致突变同步试验报告

在检索国内外文献及检测大量药食两用的天然果蔬，中药材的基础上，从中筛选出几十种有抗突变作用的可食性物质，逐步组合不同配比的组方，反复研究的组合方案，筛选出最佳配方，经特殊提取及加工制成了以山楂，枸杞，大枣，杏仁等多种原料的果茶，并经审批鉴定，制成工业产品，对试验品和工业品随机抽样做抗突变和致突变同步试验，结果该果对丝裂霉素Ｃ（ＭＭＣ）的致突变性有明显的拮抗作用，而无致突变性。     

酸角的致突变性研究

本文用组合试验评价酸角的致突变性。结果表明：酸角在Ａｍｅｓ、小鼠骨髓细胞微核和精子畸形、枯草杆菌ＤＮＡ修复试验及大肠杆菌ＷＰ２和ＷＰ２ＵＶｒＡ菌株中均为阴性反应；在大肠杆菌ＭＲ２－１０２和ＮＤ１６０菌株中为阳性反应，是酸角中的乳糖所致，ＳＣＥ频率随酸角剂量增加而稍有降低，与对照组比有显著性差异（Ｐ〈０．０５），揭示酸角可能有抗ＤＮＡ损伤作用。     

化妆品的致突变和抗突变同步快速试验分析

目的与方法: 为了快速筛选有致突变性的化妆品,采用大肠杆菌溶源性菌GY5027和对噬菌体敏感的指示菌GY4015,对29种化妆品进行致突变和抗突变同步快速试验.同时观察了急性经口试验、急性眼睛刺激试验和急性或多次皮肤刺激试验.结果:在加和不加大鼠肝脏微粒体酶(S-9)条件下,全部样品无抗突变阳性;4种样品呈现致突变阳性,其中2种至少1项急性试验阳性.经SPSS软件作等级相关分析表明,同步试验与急性试验结果相关显著(r-s＝0.392,P=0.036).结论:部分化妆品确实存在致突变性;采用致突变和抗突变同步试验可快速、大量筛选有致突变性的化妆品;急性试验阳性化妆品致突变性也较高.     

MUTAGENICANDANTIMUTAGENICDETECTIONOFSODIUMBISELENITEINVITRO

亚硒酸氢钠（ＮａＨＳｅＯ３）的致突变和抗突变作用尚无定论，本文应用稍加修改的ＳＯＳ／Ｕｍｕ显色试验对其这方面的作用作了研究。结果发现，ＮａＨＳｅＯ１剂量在０．６７－８．００ｍｇ／ｍｌ范围内，无Ｓ９时，ＳＯＳ反应的诱导率（Ｒ）有剂量一效应关系，最低诱导剂量为１．８６ｍｇ／ｍｌ：存在Ｓ９时，各剂量组的Ｒ值均接近于１。结果还揭示ＮａＨｓｅＯ３能明显抑制丝裂霉素Ｃ（ＭＭＣ）诱发的β－半乳糖苷酶（ＢＧａｓｅ）活性，其剂量－效应曲线呈抛物线。据此曲线的方程式，估得２０％、４０％和６０％抑制的剂量分别为０．４８，１．３７和３．２１ｍｇ／ｍｌ；最大有作用剂量为３．５８ｍｇ／ｍｌ，其抑制率为６１％，这些结果表明：ＮａＨＳｅＯ３可能是ＳＯＳ的直接诱导剂，同时又是ＭＭＣ诱发ＳＯＳ反应的强抑制刑，且可在无明显诱导作用的剂量（＜１．８６ｍｇ／ｍｌ）下，即可有抗ＭＭＣ的诱导作用。     

41. INHIBITION ON SOS INDUCTION BY THREE NATURAL FRUITS

Aim: As the quality of environment becomes more and more deteriorated, incidence of cancer rises steadily as a result of accumulation of carcinogenetic risk. It is impossible for people to keep away from carcinogenetic substances completely. therefore screening of edible antimutagens or anticarcinogens. from natural food such as fruit or vegetable or herbs to offset the carcinogenetic effect of carcinogens is especially important for cancer prevention. According to the traditional chinese medical theory and practice. food and medicine are equally capable for the treatment and prevention of diseases. The present study is aimed to find natural edible antimutagenic substances to improve the health improving quality of everyday food. Methods: Mutational and antimutational rapid synchronous test was employed to examine the reaction of water melon. sweet melon, strawberry, baby soybean, and baby helan bean towards the mutagenesis induced by Mitomicy (MMC) in esogenic strain GY5027. Results: Water melon. sweet melon. and strawberry inhibited mutagenesis induced by MMC in GY5027, but baby soybean and baby helan bean showed no effect. Conclusion: Water melon, sweet melon, and strawberry may have antimutagenic ability, their health improving role should be stressed in everyday life.     

Long‐term ethanol exposure causes human liver cancer cells to become resistant to mitomycin C treatment through the inactivation of bad‐mediated apoptosis

The aim of this study was to test whether long‐term ethanol consumption confers therapeutic resistance to human liver cancer patients infected with hepatitis B virus (HBV). Chronic ethanol‐treated cells were established by consecutively culturing a human hepatocellular carcinoma cell line, Hep 3B, which contains integrated HBV sequences, for 20–40 passages with or without 10 mM ethanol (designated as E20–E40 and C20–C40, respectively). Flow cytometry analysis demonstrated that a growth promoting effect of long‐term ethanol treatment was induced in the E40 cells through preferential acceleration of S‐phase in these cells. Lower protein expression levels of p16, p21/Cip1, and p27/Kip1 were detected in the ethanol‐treated E40 cells. We further demonstrated that long‐term ethanol‐treated E40 cells develop drug resistance in response to mitomycin C (MMC) treatment (>8 µM). Immunoblot analysis revealed that caspase‐8‐mediated mitochondrial apoptotic signals (such as Bad) were inactivated in the MMC‐resistant E40 cells. Immunoprecipitation experiments demonstrated that the sequestration of phosphorylated Bad (Ser‐112) through its binding with 14‐3‐3 was detected more profoundly in the MMC‐resistant E40 cells. Next, we examined the therapeutic efficacy of MMC (10 mg MMC/kg body weight, three times per week) in severe combined immunodeficient (SCID) mice bearing E40‐ and C40‐xenografted tumors. Significant reductions (>3‐fold) in tumor growth were detected in MMC‐treated C40‐xenografted mice. In vivo and in vitro studies demonstrated that AKT‐ and extracellular signal‐regulated kinase (ERK)‐mediated survival factors inhibited the Bad‐induced mitochondrial apoptotic signals that were involved in E40 tumor cells and that conferred resistance to MMC. © 2010 Wiley‐Liss, Inc.     

Anti-tumor activity of peripheral lymphocytes of tumor-bearing rats by in vitro culture

This basic study was performed to determine whether the anti-tumor effect of lymphocytes obtained from the peripheral blood of a tumor bearing host can be increased when cultured in vitro with Mitomycin C (MMC)-treated AH109A tumor cells, using interleukin-2 (IL-2). Donryu rats were used as tumor bearing hosts. The following results were obtained. Weak anti-tumor activity of lymphocytes was noted 7 days after lymphocytes were cultured in the medium to which only IL-2 was added. Anti-tumor activity was augmented by adding antigen (MMC treated tumor cells) to the above (1). Anti-tumor activity was further augmented by adding to (2) above antigen presenting cells such as intraperitoneal exudate macrophages or peripheral hole leukocytes. Anti-tumor activity of lymphocytes was detected when IL-2 and MMC treated antigen were added to the peripheral hole leukocytes containing the lymphocytes.     

普洱茶水提取物对丝裂霉素C诱发人成淋巴细胞株遗传损伤的影响

目的：探讨普洱茶水提取物对丝裂霉素C（mitomycin C,MMC）诱发的人成淋巴细胞株遗传损伤的影响。方法：以正常人成淋巴细胞株GM12593为实验材料,设计对照组、MMC作用组和普洱茶组不同条件培养细胞株。对照组用RPMI-1640正常培养,MMC作用组培养基中MMC的终浓度为0.1μg/ml,普洱茶组培养基中除含有0.1μg/ml的MMC外,还分别加入浓度为0.125、0.25、0.5和1 mg/ml的普洱熟茶和生茶水提取物。培养24 h后加入细胞松弛素B,作用28 h后制片,计数不同培养条件下的双核细胞微核率。结果：普洱熟茶在0.125和0.25 mg/ml,普洱生茶在0.125 mg/ml时,均明显降低由MMC引起的细胞微核率（P〈0.05）;普洱熟茶降低MMC诱发的细胞微核率的作用略优于普洱生茶,但两者间的差异无统计学意义。结论：一定浓度的普洱茶能降低由MMC诱发的人成淋巴细胞株的遗传损伤。     

吡柔比星和丝裂霉素C膀胱灌注预防膀胱肿瘤术后复发的临床研究

目的评价吡柔比星(THP)、丝裂霉素C(MMC)40mg、丝裂霉素C 10mg膀胱灌注预防膀胱肿瘤术后复发的疗效和安全性.方法将近期收治的86例膀胱肿瘤术后患者分为3组,分别接受THP、MMC治疗,随访12～20个月.结果THP组26例,复发2例,复发率7.7%;MMC组26例,复发3例,复发率11.5%;MMC组31例,复发11例,复发率35.5%.THP组与MMC组相比差异无显著性(P＞0.05),而THP组和40mg MMC组与MMC组相比则差异有显著性(P＜0.05).结论吡柔比星和丝裂霉素C40 mg 膀胱灌注预防膀胱肿瘤术后复发效果满意,优于丝裂霉素C 10mg膀胱灌注.     

Low thymidylate synthase expression in the primary tumor predicts favorable clinical outcome in resected gastric cancer patients treated with adjuvant tegafur

OBJECTIVE: To assess the thymidylate synthase protein (TS) in tumor cells of resected gastric cancer patients treated with adjuvant tegafur (TG), we reviewed the outcome of 94 randomized patients treated either with adjuvant TG plus mitomycin C (MMC) or MMC alone. METHODS: TS was determined in 76 out of 94 patients, previously randomized to receive adjuvant TG, 500 mg/m(2)/day p.o. for 6 months plus four courses of MMC, 10- 20 mg/m(2) i.v. every 6 weeks or MMC alone. An immunohistochemical assessment with the monoclonal antibody TS-106 was performed. RESULTS: Low TS was observed in 38 patients (20 treated with TG-MMC and 18 with MMC) and high TS in the other 38 patients (21 treated with TG-MMC and 17 with MMC). After 10 years' median follow-up time, 61% of adjuvant TG-MMC patients and 43% of MMC patients were alive and disease-free. Disease-free survival and overall survival were significantly better for patients treated with TG-MMC compared to MMC adjuvant (p = 0.0277 and p = 0.05), and low-TS compared to high-TS patients (p = 0.0184 and p = 0.0198). In 38 low-TS patients we observed an 83% chance to be disease-free in TG-MMC-treated patients and 55% in MMC-treated patients (p = 0.04). CONCLUSIONS: A low TS level determines a subset of patients that may benefit from adjuvant oral TG when added to MMC showing a 5-year cure rate of more than 80%.     

Multihospital randomized study on adjuvant chemotherapy with mitomycin C and futraful or UFT in gastric cancer--(Part 2). Comparisons between futraful and UFT. North Kyushu Co-operative Study Group for Cancer Chemotherapy

A comparative multicenter study was carried out on postoperative adjuvant chemotherapy on 243 gastrectomy patients from October 1983 to December 1985. The medications were assigned at random to 2 groups of patients by the envelope method: group A (administered 20 mg of MMC on the day of surgery and 10 mg of MMC on the day after surgery, and started on tegafur at a dose of 600 mg/day 2 weeks after surgery); group B (similarly treated with MMC and started on UFT at a dose of 600 mg/day 2 weeks after surgery). Further, patients presenting in stage IV were administered MMC intravenously every 3 months, starting 4 weeks after surgery. The final drug effect evaluations were made in 210 patients excluding 33 dropouts (13.6%). The incidence of side effects was somewhat higher in group B than in group A, but their frequency was low and no serious symptoms were observed. For patients with poorly differentiated adenocarcinoma and were carried out curative operation, the 3-year survival rate was significantly higher in group B than in group A.     

Comparative study of tegafur (p.o.) and mitomycin C (MMC) plus tegafur in cases of unresectable gastric cancer

Sixteen cases of gastric cancer with liver metastases (H2 less than) as group A, fifteen with distant peritoneal metastases (P2 less than) as group B, twenty-five with distant lymph nodes metastases (N3 less than) as group C, twenty-eight with two prognostic factors as group D, and seven with three factors as group E were treated. A1, B1, C1, D1 and E1 groups were treated with mitomycin C (MMC) (4 mg) + tegafur (FT) (400 mg) (i.v.-weekly) and A2, B2, and C2 group with FT (600 mg) (p.o.-daily). Partial response in the evaluable cases was found in 7 cases and a total response rate was 11.7%. The 50% survival interval was as follows; 8.5 months (M) in A1 group, 10.0 M in A2, 7.2 M in B1, 2.3 M in B2, 10.4 M in C1, 14.2 M in C2, 5.2 M-7.2 M in D1, and 3.1 M in E1. In the cases with H2 less than or N3 less than, FT (p.o.) and with P2 less than, MMC + FT (i.v.) were effective respectively. Chemotherapeutic effect was better in A1 and A2 group from the aspects of prognostic factors and in the cases of poorly differentiated adenocarcinoma from histological types than in B1 and B2 group and well or moderately differentiated adenocarcinoma.     

Postoperative chemo-endocrine treatment with mitomycin C, tamoxifen, and UFT is effective for patients with premenopausal estrogen receptor-positive stage II breast cancer. Nishinihon Cooperative Study Group of Adjuvant Therapy for Breast Cancer

The effectiveness of combining mitomycin C (MMC), tamoxifen (TAM), and 1-(2-tetrahydrofuryl)-5-fluorouracil (tegafur) was evident in patients with estrogen receptor-positive (ER+) breast cancers. UFT, an oral preparation of tegafur and uracil at a molar ratio of 1:4, was reported to have higher antitumor effects than tegafur alone for patients with breast cancer. Therefore, the combined chemotherapy of MMC, TAM and UFT may possibly be effective for breast cancer. From 1988 to 1991. we studied the effects of postoperative adjuvant therapy for Japanese women with stage 11 breast cancer, all seen at 71 institutions in western areas of Japan. Five hundred and ninety four patients with stage II primary breast cancer who had undergone curative surgery, including total mastectomy and axillary lymph node dissection, were enrolled. On the day of surgery, each patient was given 13 mg/m2 of MMC intravenously. Patients with ER+ tumors were then assigned to group A or group B. Group A received 30 mg/day of TAM given orally from postoperative 2 weeks, for 2 years. Group B was additionally given an oral dose of 300 mg/day of UFT for 2 years, given concomitantly with 30 mg/day of TAM. Patients with ER- tumors were assigned to group C or group D. Group C were prescribed 300 mg/day of UFT, orally, from postoperative 2 weeks for 2 years, and group D were additionally given an oral dose of 30 mg/day of TAM together with 300 mg/day of UFT. There were no differences among the groups regarding prognostic factors or doses of MMC and TAM in ER+ patients and MMC and UFT in ER- patients. Toxicity rates for leukopenia, anorexia, and nausea/vomiting were higher in group B than in group A patients. There were no statistical differences in the overall survival and disease-free survival times between groups A and B, or groups C and D, for all eligible cases. In a retrospective subgroup analysis using Bonferroni's adjustments, the additional effect of UFT on the combined treatment of MMC and TAM lengthened the disease-free survival time for patients with premenopausal ER+ cancers (corrected P value by Bonferroni's adjustments <0.05). Multivariate analysis showed that effects of the combined treatment of MMC, TAM, and UFT was significantly related to the menopausal status (P<0.01). Our findings show that postoperative ingestion of MMC, TAM, and UFT was effective for patients with premenopausal ER+ stage II breast cancer.     

丝裂霉素膀胱灌注预防浅表性膀胱肿瘤复发的远期疗效观察

目的 观察丝裂霉素预防浅表性膀胱肿瘤复发的远期疗效。方法 将１９８０年１月￣１９９１年１２月的８６例浅生膀胱肿瘤患者分为丝裂霉素组（手术＋丝裂霉素灌注）和对照组（单纯手术）,对两组的肿瘤术后复发率、病理分期和细胞分级进行对比观察。结果 丝霉素组７１例,对照组１５例,术后肿瘤复发率分别为４０．８％和６６．７％。丝裂霉素组在术后６月、１２月、２４月、３６月和６０月以上的无瘤率分别为８５．９％,７６．１