EXPERIMENTAL STUDY OF ANTI－TUMOR EFFECT OF ELECTROCHEMICAL THERAPY ALONE OR IN COMBINATION WITH RADIOTHERAPY

徐晓娜，肖卫群ＥＸＰＥＲＩＭＥＮＴＡＬＳＴＵＤＹＯＦＡＮＴＩ－ＴＵＭＯＲＥＦＦＥＣＴＯＦＥＬＥＣＴＲＯＣＨＥＭＩＣＡＬＴＨＥＲＡＰＹＡＬＯＮＥＯＲＩＮＣＯＭＢＩＮＡＴＩＯＮＷＩＴＨＲＡＤＩＯＴＨＥＲＡＰＹ￥ＸｕＸｉａｏｎａ；ＸｉａｏＷｅｉｑｕｎ（Ｄｅ...     

CLONING AND SEQUENCING OF IMMUNOGLOBULINVARIABLE－REGION GENE OF A MONOCLONALANTIBODY SPECIFIC FOR HUMANHEPATOCARCINOMA

ＣＬＯＮＩＮＧＡＮＤＳＥＱＵＥＮＣＩＮＧＯＦＩＭＭＵＮＯＧＬＯＢＵＬＩＮＶＡＲＩＡＢＬＥ－ＲＥＧＩＯＮＧＥＮＥＯＦＡＭＯＮＯＣＬＯＮＡＬＡＮＴＩＢＯＤＹＳＰＥＣＩＦＩＣＦＯＲＨＵＭＡＮＨＥＰＡＴＯＣＡＲＣＩＮＯＭＡＹａｎｇＰｉｎｇ杨萍；ＧａｏＬｅｉ高...     

PREVENTIVE DETECTION OF FUNGI AND MYCOTOXINS IN CORN FROM HIGH RISKAREA OF ESOPHAGEAL CANCER IN CIXIAN COUNTY

ＰＲＥＶＥＮＴＩＶＥＤＥＴＥＣＴＩＯＮＯＦＦＵＮＧＩＡＮＤＭＹＣＯＴＯＸＩＮＳＩＮＣＯＲＮＦＲＯＭＨＩＧＨＲＩＳＫＡＲＥＡＯＦＥＳＯＰＨＡＧＥＡＬＣＡＮＣＥＲＩＮＣＩＸＩＡＮＣＯＵＮＴＹＺｈａｎｇＸｉａｎｇｈｏｎｇ；张祥宏；ＸｉｅＴｏｎｇｘｉｎ；谢同...     

THE EXPRESSION OF C－MYC AND N－RAS ONCOGENES IN HUMAN HEPATOCELLULAR CARCINOMA－AN IN SITU HYBRIDIZATION STUDY ON PARAFFIN EMBE

ＴＨＥＥＸＰＲＥＳＳＩＯＮＯＦＣ－ＭＹＣＡＮＤＮ－ＲＡＳＯＮＣＯＧＥＮＥＳＩＮＨＵＭＡＮＨＥＰＡＴＯＣＥＬＬＵＬＡＲＣＡＲＣＩＮＯＭＡ－ＡＮＩＮＳＩＴＵＨＹＢＲＩＤＩＺＡＴＩＯＮＳＴＵＤＹＯＮＰＡＲＡＦＦＩＮＥＭＢＥＤＤＥＤＴＩＳＳＵＥＳＥＣＴＩＯＮ...     

THE IN VITRO POTENTIATION OF LAK CELL CYTOTOXICITY IN CANCER AND AIDS PATIENTS INDUCED BY F3—A FRACTIONATED EXTRACT OF ASTRAG

ＴＨＥＩＮＶＩＴＲＯＰＯＴＥＮＴＩＡＴＩＯＮＯＦＬＡＫＣＥＬＬＣＹＴＯＴＯＸＩＣＩＴＹＩＮＣＡＮＣＥＲＡＮＤＡＩＤＳＰＡＴＩＥＮＴＳＩＮＤＵＣＥＤＢＹＦ３—ＡＦＲＡＣＴＩＯＮＡＴＥＤＥＸＴＲＡＣＴＯＦＡＳＴＲＡＧＡＬＵＳＭＥＭＢＲＡＮＡＣＥＵＳＣｈｕ...     

SPECIFIC UPTAKE OF MONOCLONAL ANTIBODY-CONJUGATED METHOTREXATE BY HUMAN LYMPHOCYTIC LEUKEMIC B CELLS

ＳＰＥＣＩＦＩＣＵＰＴＡＫＥＯＦＭＯＮＯＣＬＯＮＡＬＡＮＴＩＢＯＤＹＣＯＮＪＵＧＡＴＥＤＭＥＴＨＯＴＲＥＸＡＴＥＢＹＨＵＭＡＮＬＹＭＰＨＯＣＹＴＩＣＬＥＵＫＥＭＩＣＢＣＥＬＬＳＺｈｕＺｈｅｎｐｉｎｇ１朱祯平ＹａｎｇＣｈｕｎｚｈｅｎｇ１杨纯正Ｔａｒｕ...     

THE BLOCKING EFFECTS OF GLYCYRRHIZE URALENSIS AND CHELIDONIUM MAJUS ON MUTAGENESIS INDUCED BY AFLATOXIN B1

ＴＨＥＢＬＯＣＫＩＮＧＥＦＦＥＣＴＳＯＦＧＬＹＣＹＲＲＨＩＺＥＵＲＡＬＥＮＳＩＳＡＮＤＣＨＥＬＩＤＯＮＩＵＭＭＡＪＵＳＯＮＭＵＴＡＧＥＮＥＳＩＳＩＮＤＵＣＥＤＢＹＡＦＬＡＴＯＸＩＮＢ１ＳｈｉＧｕｉｚｎｉ；史桂芝；ＪｉＸｉｎｈｕａ；纪新华；ＬｉａｎｇＹ...     

QUALITATIVE STUDY OF SIALOMUCINS CHANGES DURING N－METHYL－N－NITROSOUREA－INDUCED C OLONIC CARCINOGENESIS IN MICE

ＱＵＡＬＩＴＡＴＩＶＥＳＴＵＤＹＯＦＳＩＡＬＯＭＵＣＩＮＳＣＨＡＮＧＥＳＤＵＲＩＮＧＮ－ＭＥＴＨＹＬ－Ｎ－ＮＩＴＲＯＳＯＵＲＥＡ－ＩＮＤＵＣＥＤＣＯＬＯＮＩＣＣＡＲＣＩＮＯＧＥＮＥＳＩＳＩＮＭＩＣＥＷａｎｇＱｉａｎｇ王强；ＷａｎｇＹｕａｎｈｅ王元和；...     

APOPTOSIS OF TUMOR CELLS IN LECTIN－DEPENDENTLYMPHOKINE－ACTIVATED KILLER CELLMEDIATED CYTOTOXICITY

ＡＰＯＰＴＯＳＩＳＯＦＴＵＭＯＲＣＥＬＬＳＩＮＬＥＣＴＩＮ－ＤＥＰＥＮＤＥＮＴＬＹＭＰＨＯＫＩＮＥ－ＡＣＴＩＶＡＴＥＤＫＩＬＬＥＲＣＥＬＬＭＥＤＩＡＴＥＤＣＹＴＯＴＯＸＩＣＩＴＹＤｏｎｇＨａｉｄｏｎｇ董海东；ＸｉｎｇＲｏｎｇ邢嵘；ＧｕｏＬｉａｎｙｉｎ...     

A MICROCOMPUTER PROGRAM FOR CALCULATINGTHE CONFIDENCE INTERVALS OF SURVIVAL PROBABILITY IN MEDICAL FOLLOW－UP STUDIES

ＡＭＩＣＲＯＣＯＭＰＵＴＥＲＰＲＯＧＲＡＭＦＯＲＣＡＬＣＵＬＡＴＩＮＧＴＨＥＣＯＮＦＩＤＥＮＣＥＩＮＴＥＲＶＡＬＳＯＦＳＵＲＶＩＶＡＬＰＲＯＢＡＢＩＬＩＴＹＩＮＭＥＤＩＣＡＬＦＯＬＬＯＷ－ＵＰＳＴＵＤＩＥＳＸｉａｎｇｙｏｎｇｂｉｎｇ项永兵；Ｇａｏｙｕ...     

双功能抗体介导胃癌杀伤效应的研究

目的 以化学方法将抗CD3及抗三硝基苯（TNP）的单克隆抗体（McAb)交联为双功能抗体。利用此双功能抗体介导人外周血淋巴细胞（PBLS）对于不同抗胃癌McAb所针对的胃癌细胞进行体内外杀伤试验。方法 利用化学交联法制备双功能抗体,以酶联免疫吸附试验(ELISA）测定McAbs活性,以ELISA桥联法、琼脂糖免疫双扩散及十二烷基磺酸钠－降丙烯酰胺凝胶电泳（SDS－PAGE）对于双功能抗体进行鉴定,以细胞杀伤试验及肿瘤生长抑制实验检测双功能抗体作用。结果 将抗CD3及抗TNP McAb交联为双功能抗体,此双功能抗体可介导PBLS对于不同抗胃癌McAb所针对的胃癌细胞进行体内外杀伤试验。结论 此双功能抗体在肿瘤生物免疫治疗中具有应用前景。     

Alteration of the daunorubicin-triggered sphingomyelin-ceramide pathway and apoptosis in MDR cells: influence of drug transport abnormalities

We have previously shown that in myeloid leukemic cells, daunorubicin (DNR) induces apoptosis via the activation of the sphingomyelin-ceramide pathway. We have now investigated sphingomyelin (SM) hydrolysis, ceramide generation, and apoptosis in vincristine-selected multidrug resistant (MDR) HL-60 cells (HL-60/Vinc), compared with their parental counterparts. We show that DNR triggers the SM cycle (stimulation of neutral sphingomyelinase, SM hydrolysis, and ceramide generation) and apoptosis in both parental and MDR cells, when used at isotoxic doses (ie., 1 and 100 microM for HL-60 and HL-60/Vinc, respectively). However, in MDR cells treated with either 10 microM DNR or 1 microM DNR in association with the P-glycoprotein (P-gp) blocker verapamil (treatment conditions which yield an intracellular DNR concentration similar to that achieved with 1 microM in the parental cells), we were unable to detect SM hydrolysis, ceramide generation and apoptosis. This implies that inhibition of the DNR-induced SM cycle in MDR cells is not directly related to P-gp. We have also investigated the influence of intracellular drug localization on the DNR-induced SM-cycle in HL-60/Vinc cells. In these cells, DNR at 10 microM is mainly localized in cytoplasmic vesicles, while the drug is diffusely distributed when used at 100 microM. A diffuse distribution pattern was also observed when MDR cells were treated with 1 microM DNR in association with the cyclosporine derivative PSC-833, but not with verapamil. In parallel, PSC-833, but not verapamil, restored the induction of the SM cycle and the apoptotic potential of DNR, and markedly increased drug cytotoxicity in MDR cells. Our results suggest that altered intracellular drug transport plays an important role in limiting ceramide generation and cell death in MDR cells.     

Compared cytotoxicity effects of five anticancer drugs on human (HBL) and mouse (B16) melanoma cells in vitro

In order to assess the potential interest of replacing the murine cell lines by human cell lines for in vitro cytotoxic assays, the sensitivity and the selectivity of the murine B16 and the human HBL melanomas to five chemotherapeutic drugs were investigated in vitro. The cytotoxicities of Melphalan, Daunorubicin (DNR), Hexamethylmelamine (HMM), Hydroxymethylpentamethylmelamine (HMPMM), and Dihydroxymethyltetramethylmelamine (DHTMM), 2 HMM derivatives, were measured in the two cell lines using two different techniques: reduction of a tetrazolium derivative (MTT) and tritiated thymidine uptake into DNA. The cytotoxicity at inhibitory concentration 50 (IC50) was determined after one hour as well as after 2 days exposure of cell after one hour as well as after 2 days exposure of cells to each drug. The results indicate that the HBL human melanoma was generally more sensitive to Melphalan and DHTMM than the B16 murine melanoma cells as far as the IC50 was concerned. In contrast, no difference of sensitivity was found to DNR and DHTMM. HMM was found to be inactive in both cell lines. The analysis of variance on IC50 values showed that the sensitivity of murine and human melanoma cell lines to drugs was statistically different. Despite the identical selectivity of the two cell lines, two promising observations can be made as far as the comparison of the two cell lines is concerned: 1) the higher sensitivity of HBL human cell line to Melphalan in the in vitro assays and 2) the slightly lower sensitivity of HBL to DNR, a drug without clinical activity against human melanoma.     

Do P-glycoprotein and major vault protein (MVP/LRP) expression correlate with in vitro daunorubicin resistance in acute myeloid leukemia?

Two proteins that have been correlated with the occurrence of multidrug resistance in acute myeloid leukemia (AML) are P-glycoprotein (Pgp) and the major vault protein (Mvp/LRP). With the purpose of further quantifying the potential contributions of Pgp-mediated drug efflux and Mvp/LRP to drug resistance in AML we have investigated whether the transport function of Pgp and the expression of Mvp/LRP correlated with the accumulation of daunorubicin (DNR) and the in vitro resistance to DNR cytotoxicity (LC50 by MTT assay) in AML cells. In de novo adult AML, the steady-state DNR accumulation (in pmol/10(6) cells) correlated with Pgp activity or expression, whereas the LC50 for DNR did not correlate with Pgp activity (measured as the modulation of rhodamine 123 or DNR accumulation by the Pgp inhibitor PSC833) or Pgp expression (measured by flow cytometry with the MRK-16 antibody). The contribution of MRP1 expression to a reduced DNR accumulation seems minor compared to Pgp. In addition, the modulation of the DNR LC50 by PSC833 did not correlate with Pgp protein or activity. The steady-state DNR accumulation showed no correlation with the DNR LC50. The Mvp/LRP expression (immunocytochemical staining) did neither correlate with DNR accumulation nor with the DNR LC50. A significant negative correlation was seen between the Mvp/LRP immunocytochemical staining and Pgp activity, indicating that both markers define (partially) different populations. In conclusion, it is shown that Pgp function, but not Mvp/LRP or MRP1 expression correlate with a low steady-state DNR accumulation in de novo AML. The Pgp activity does, however, not predict the DNR sensitivity in AML measured as in vitro DNR LC50 with an MTT-based assay. The reason for that seems to be that a low DNR accumulation may not be the most important factor in determining the LC50. While the clinical usefulness of these drug resistance tests remains to be proven they do not seem to provide as yet a straightforward explanation for the major cause(s) of clinical chemotherapy failure.     

Anthracycline immunoconjugates prepared by a site-specific linkage via an amino-dextran intermediate carrier

Anthracycline, either daunomycin or doxorubicin, was site specifically attached to the carbohydrate moiety of a monoclonal anticarcinoembryonic antigen antibody by using amino-dextran as the intermediate carrier. The reaction resulted in an immunoconjugate that contains approximately 20 to 25 molecules of drug per molecule of immunoglobulin G. Flow-cytometric studies revealed the retention of the antibody-binding activity. The immunoconjugate was cytotoxic to the target cells, as examined by the 75selenomethionine incorporation studies, and remained efficient for targeting a human colonic tumor (GW-39) in the nude mouse model. The conjugate possessed a greater antitumor activity against the subcutaneous tumor than either the free drug or an irrelevant antibody conjugate, and it was well tolerated by the animals at a much higher dose level than was the unconjugated drug.     

Effect of O6-methylguanine-DNA methyltransferase gene activity on repair in human cells transformed by a regulatable ada gene

To assess the biological role of DNA methylation at the O6 position of guanine (O6MeG) a human cell line was created that contains a regulatable gene of the O6MeG-DNA methyltransferase (MT), a repair activity that removes O6MeG adducts from the DNA. MT-deficient HeLa MR cells were transformed with an SV40-based expression vector in which the bacterial MT gene ada was put under the control of a glucocorticoid-inducible MMTV promoter. In response to dexamethasone (Dex), pSV MTV ada cells actively accumulated MT protein to reach a constant level after 10-12 h of approximately 15,000 MT molecules per cell. Co-induction by Dex and 12-O-tetradecanoylphorbol-13-acetate (TPA) further accelerated this synthesis approximately 2-fold and, as a result, higher final MT levels were achieved. The inducers were added to exponentially growing cells either before or at the time of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) exposure and the kinetics of MT synthesis was studied. MNNG affected in a dose-dependent manner (i) the loss of the pre-existing MT activity; (ii) the lag before newly synthesized MT appeared; (iii) the final level of MT accumulated by the cells; and (iv) to a lesser extent the rate of MT synthesis. In cells with a down-regulated MT gene (no inducer) even small MNNG doses lead to an irreversible loss of the pre-existing MT activity, i.e. to incomplete repair, whereas an up-regulated MT gene supported the restoration of a pool of active MT molecules in the cells, i.e. an O6MeG repair that has gone to completion. Hence, effective O6MeG repair relies not only on the pre-existing MT level, but depends to an even greater extent on the state of expression of the MT gene. The activity of the MT gene also correlated with cell survival, which confirms our earlier finding that O6MeG adducts are cytotoxic for the cell.     

Methimazole as a protectant against cisplatin-induced nephrotoxicity using the dog as a model

Dexniguldipine-HCl (DNIG) — a prospective clinical modulator of p170-glycoprotein (pgp170)-mediated multidrug resistance (MRD) — was evaluated in a drug-accumulation assay in MDR murine leukemia cell strain F4-6RADR expressing pgp170. The compound elevated low accumulation of either doxorubicin (DOX), daunorubicin (DNR), or mitoxantrone (MITO) in resistant F4-6RADR cells to the very levels observed in drug-sensitive F4-6 precursor cells. In parallel with the increase in DNR content (F4-6RADR, solvent: 303±27 pmol/mg protein; DNIG (3.3 mol/l): 1,067±174 pmol/mg protein; F4-6P, solvent: 948±110 pmol/mg protein;n=8–9, SEM), the amount of DNR tightly bound to the acid precipitate pellet obtained from F4-6RADR (i.e., protein, DNA, RNA) increased 3.9-times to the levels observed in sensitive F4-6 cells. The main pyridine metabolite of DNIG displayed similar activity. Concentration-response analysis revealed that DNIG and R,S-verapamil (VER) induced 100% reversal of the DNR accumulation shortage associated with the MDR phenotype but DNIG was 8 times more potent than VER (50% inhibitory concentration (IC₅₀), 0.73 vs 5.4 mol/l). In keeping with the accumulation assay, DNIG was about 10 times more potent than VER in sensitizing F4-6RADR cells to the cytostatic and cytotoxic effects of DNR in proliferation assays. In conclusion, DNIG is a potent in vitro modulator, improving (a) the accumulation of anthracycline-like cytostatics, (B) drug access to cellular binding sites, and (c) the cytostatic action of DNR in F4-6RADR leukemia cells of the MDR phenotype.Abbreviations DOX doxorubicin - CSA cyclosporin A - DMSO dimethylsulfoxide - DNIG dexniguldipine-HCl - DNR daunorubicin - MDR multidrug resistance - MITO mitoxantrone - pgp170 permease glycoprotein 170 - VER R.S.-verapamil Dexniguldipine-HCl is the proposed INN for compound B859-35, the R-enantiomer of niguldipine. Segments of this work have been reported in the abstract form     

Idarubicin overcomes P-glycoprotein-related multidrug resistance: comparison with doxorubicin and daunorubicin in human multiple myeloma cell lines.

The clinical utility of anthracyclines like doxorubicin (DOX) and daunorubicin (DNR) for treatment of multiple myeloma (MM) is limited by the occurrence of multidrug resistance (MDR). Highly lipophilic anthracyclines like idarubicin (IDA) might circumvent MDR and thereby enhance chemotherapeutic efficacy. To determine the efficacy of IDA in myeloma cells, the pharmacokinetics and cytotoxicity of IDA and its major metabolite idarubicinol (IDAol) were compared with those of DNR, DOX, and doxorubicinol (DOXol) in the cell line RPMI 8226-S and two MDR sublines (8226-R7 and 8226-Dox40) that overexpress the drug transporter P-glycoprotein (Pgp). Cytotoxicity assays using MTT (viability) or annexin V (apoptosis) showed a 10-50-fold higher potency of IDA compared with DNR or DOX in the MDR variant cell lines. The difference in cytotoxicity was lower in the sensitive parental cell line (3-fold). These results are explained by a better intracellular uptake of IDA compared to DNR in resistant 8226 cell lines. The Pgp-inhibitor verapamil affected IDA uptake only in the most resistant cell line 8226-Dox40. This indicates that IDA is less sensitive than DNR to transport-mediated MDR. IDAol was at least 32-fold more cytotoxic than DOXol, and more susceptible to Pgp transport than IDA. These studies demonstrate that the efficacy of IDA in MDR MM cell lines is superior to that of DOX or DNR, and that IDA may become an important drug in the treatment of MM, especially in refractory disease.     

Pharmacokinetic and cytotoxic studies of pegylated liposomal daunorubicin

Pegylated liposomes have been studied for nearly two decades. However, fewer pharmacological studies about its application in daunorubicin (DNR) than those in doxorubicin have been reported. In order to conduct a complete pharmacokinetic study, radiolabeled DNR was encapsulated in pegylated liposomes. Its in vitro drug release kinetics was determined to be in a slow manner, which was reflected in its cytotoxic effect on four cell lines. The lethal dose, plasma pharmacokinetics as well as tissue distribution of the formulation were evaluated in comparison with free DNR. The results revealed that liposomal daunorubicin significantly reduced the toxicity of the drug, with a half lethal dose of 29.35 mg/kg, compared with 5.45 mg/kg for free drug. Pharmacokinetic study of liposomal DNR demonstrated a slower clearance rate, an elevated area under the concentration–time curve, as well as increased half-lives compared to free drug. In addition, an altered tissue distribution of liposomal DNR was observed, with lower cardiac accumulation. Taken together, pegylated liposome-loaded DNR may be a promising anticancer drug and worth further therapeutic study.     

Overcoming effect of antibody against rat alpha-fetoprotein (AFP) on the growth of daunorubicin-resistant mutant rat ascites hepatoma cell line AH66

A daunorubicin (DNR)-resistant variant (AH66DR) of alpha-fetoprotein (AFP)-producing rat ascites hepatoma AH66 was established. AH66DR was 169 times more resistant to DNR than AH66. A growth-inhibitory effect due to the decrease in sugar uptake following treatment of the cell lines with specific antibody against highly purified rat AFP was also observed. Pre-incubation of both lines with antibody prior to the measurement of 2-deoxy-D-glucose (2dG) uptake resulted in a 45-55% decrease in 2dG uptake compared to control cells with a 2-fold reduction of Vmax value while the Km remained unchanged. Upon pre-incubating AH66DR with antibody, an increased intracellular level of DNR concentration with reduction of efflux rate was observed. A significantly superior cytotoxic effect against AH66DR, which reached almost the same level as the cytotoxic effect of DNR against AH66, was found, as compared to various other controls when tumor cells were cultured with a mixture of antibody and DNR. Our in vitro results suggest that the DNR-resistance of the AFP-producing tumor cells may be overcome if the cells are treated with specific antibody.