ObjectiveTo investigate the bioeffects of extremely low frequency (ELF) magnetic field (MF) (50 Hz, 400μT) and magnetic nanoparticles (MNPs) via cytotoxicity and apoptosis assays on PC12 cells.
MethodsMNPs modified by SiO2 (MNP-SiO2) were characterized by transmission electron microscopy (TEM), dynamic light scattering and hysteresis loop measurement.PC12 cells were administrated with MNP-SiO2 with or without MF exposure for 48 h. Cytotoxicity and apoptosis were evaluated with MTT assay and annexin V-FITC/PI staining, respectively. The morphology and uptake of MNP-SiO2 were determined by TEM. MF simulation was performed by Ansoft Maxwell based on the finite element method.
ResultsMNP-SiO2 were identified as~20nm (diameter) ferromagnetic particles. MNP-SiO2reduced cell viability in a dose-dependent manner. MF also reduced cell viability with increasing concentrations of MNP-SiO2. MNP-SiO2 alone did not cause apoptosis in PC12 cells; instead, the proportion of apoptotic cells increased significantly under MF exposure and increasing doses of MNP-SiO2. MNP-SiO2 could be ingested andthen cause a slight change in cellmorphology.
ConclusionCombined exposure of MF and MNP-SiO2 resulted in remarkable cytotoxicity and increased apoptosis in PC12 cells. The results suggested that MF exposure couldstrengthen the MF of MNPs, which may enhance the bioeffects of ELF MF.

ObjectiveTo characterize two strains of street rabies virus (RABV) isolated from the brain tissue of cattle from Inner Mongolia. Differences in the histopathological and ultrastructural changes in the brain tissue of infected mice were determined to reveal variation in the pathogenesis of infection between street rabies virus strains. MethodsTen-day-old mice were intracranially inoculated with one of three virus strains and brain tissue harvested when the mice were moribund. Various histopathological and ultrastructural markers of disease were then compared between the groups. ResultsInfection with the street virus strain CNM1101C resulted in severe neuronal dendrites damage, but only mild cell apoptosis, T lymphocyte infiltration and microglial activation. Infection with the other street virus strain,CNM1103C, was characterized by cell apoptosis, T lymphocyte infiltration and microglial activation as well as dendrites damage. However, in comparison, infection with the attenuated virus strain CTN caused severe T lymphocyte infiltration, microglial activation and cell apoptosis, but left the neuronal dendrites intact. ConclusionThe two street rabies virus strains isolated from cattle from Inner Mongolia had different levels of virulence and caused distinct pathological changes in infected mice. Therefore, we concluded that different pathogenic mechanisms exist between different RABV strains.

Objective A subcutaneous transplantation tumor model of human HT-29 cells in nude mice was established to evaluate anticarcinogenic activities, and the apoptosis-regulated mechanism effect of aqueous extract of fermented wheat germ with Lactobacillus plantarum dy-1 (LFWGE).
Methods The HT-29 cells were transplanted via subcutaneous injection of 1×107 cells into the right flank of each nude mouse. Then, nude mice were treated for 30 d with LFWGE (high-dose 2 g/kg/d;low-dose 1 g/kg/d) and for 7 d with 5-fluorouracil (5-FU, 25 mg/kg/d) by gavage and intraperitoneal injection, respectively. An inhibition of tumor growth was observed.
Results Tumor volume and weights decreased significantly in both groups of nude mice treated with LFWGE. In addition, the cell apoptosis rate of the LFWGE group (2 g/kg/d, 60.1%±4.4%; 1 g/kg/d, 58.6%±6.9%) was significantly higher than that of the control group (11.5%±1.6%) and 5-FU group (32.1%±3.5%) as measured by the TUNEL assay. Moreover, the real-time fluorescent quantitative PCR and Western blot method further confirmed these enhancing apoptosis and growth inhibition effects. The involvement of LFWGE in inducing apoptosis was confirmed by the expression of Bax, Bcl-2, Caspase-3, and CyclinD1.
Conclusion The results showed that LFWGE could induce subcutaneous transplantation tumor apoptosis in nude mice and could be as a natural nutrient supplements or chemopreventive agent in the treatment of human colon cancer.

Objective To explore the effects of resveratrol-induced apoptosis and autophagy in T-cell acute lymphoblastic leukemia (T-ALL) cells and potential molecular mechanisms.
Methods The anti-proliferation effect of resveratrol-induced, apoptosis and autophagy on T-ALL cells were detected by using MTT test, immunofluorescence, electronic microscope, and flow cytometry, respectively. Western blotting was performed for detecting changes of apoptosis-associated proteins, cell cycle regulatory proteins and state of activation of Akt, mTOR, p70S6K, 4E-BP1, and p38-MAPK.
Results Resveratrol inhibited the proliferation and induced apoptosis and autophagy in T-ALL cells in a dose and time-dependent manner. It also induced cell cycle arrest at G0/G1 phase via up regulating cyclin-dependent kinase (CDK) inhibitors p21 and p27 and down regulating cyclin A and cyclin D1. Western blotting revealed that resveratrol significantly decreased the expression of antiapoptotic proteins (Mcl-1 and Bcl-2) and increased the expression of proapoptotic proteins (Bax, Bim, and Bad), and induced cleaved-caspase-3 in a time-dependent manner. Significant increase in ratio of LC3-II/LC3-I and Beclin 1 was also detected. Furthermore, resveratrol induced significant dephosphorylation of Akt, mTOR, p70S6K, and 4E-BP1, but enhanced specific phosphorylation of p38-MAPK which could be blocked by SB203580. When autophagy was suppressed by 3-MA, apoptosis in T-ALL cells induced by resveratrol was enhanced.
Conclusion Our findings have suggested that resveratrol induces cell cycle arrest, apoptosis, and autophagy in T-ALL cells through inhibiting Akt/mTOR/p70S6K/4E-BP1 and activating p38-MAPK signaling pathways. Autophagy might play a role as a self-defense mechanism in T-ALL cells treated by resveratrol. Therefore, the reasonable inhibition of autophagy in T-ALL cells may serve as a promising strategy for resveratrol induced apoptosis and can be used as adjuvant chemotherapy for T-ALL.

ObjectiveInactivated Sendai virus particle [hemagglutinating virus of Japan envelope (HVJ-E)] has a potential oncolytic effect due to its ability to induce apoptosis in tumor cells. However, the molecular mechanism of apoptosis induction in cancer cellsmediated by HVJ-E has not been fully elucidated.This paper aims to investigate the underlying mechanism of apoptosis induction by HVJ-E in prostate cancer cells (PC3). MethodsPC3 cells were treated with HVJ-E at various MOI, and theninterferon-β (IFN-β) production, and the cell viability and apoptosis were detected by ELISA, MTT-based assay and flow cytometry, respectively. Next, the roles of Jak-Stat, MAPK and Akt pathways played in HVJ-E-induced apoptosis in PC3 cells were analyzed by immunoblot assay. To further evaluate the cytotoxic effect of HVJ-E on PC3 cells, HVJ-E was intratumorally injected into prostate cancers on BALB/c-nude mice, and the tumor volume was monitored for 36 days. ResultsHVJ-E induced IFN-β production and activatedJak-Stat signaling pathway, which resulted in the activation of caspase-8, caspase-3, and PARP in PC3 prostate cancer cells post HVJ-E treatment. Furthermore, we observed for the first time that p38 and Jnk MAPKs in PC3 cells contributed to HVJ-E-induced apoptosis. In addition,intratumoralHVJ-E treatmentdisplayed a directinhibitoryeffect in anin vivo BALB/cnude mouseprostate cancermodel. ConclusionOur findingshaveprovided novel insights into the underlying mechanismsby whichHVJ-E induces apoptosisin tumor cells.

Objective To investigate the role of extracellular signal-regulated kinase (ERK) in apoptosis of human colon cancer (HCT116) cells.
Methods After the HCT116 cells were pretreated with specific ERK inhibitor (U0126) or specific siRNA and exposed to 10 mmol/L sodium butyrate (NaBT) for 24 h, their apoptosis was detected by flow cytometry, levels of SphK2 and ERK protein were measured by Western blot, and translocation of SphK2 was assayed by immunofluorescence microscopy.
Results The U0126 and siRNAs specific for SphK2 blocked the export of SphK2 from nuclei to cytoplasm and increased the apoptosis of HCT116 cells following NaBT exposure. Over-expression of PKD decreased NaBT-induced apoptosis of HCT116 cells, which was reversed by U0126. Furthermore, transfection of HCT116 cells with constitutively activated PKD plasmids recovered the U0126-blocked export of SphK2.
Conclusion ERK regulates the export of SphK2 and apoptosis of HCT116 cells by modulating PKD. Modulation of these molecules may help increase the sensitivity of colon cancer cells to the physiologic anti-colon cancer agent, NaBT.

TNFα1型受体敲除对小鼠急性后肢缺血的作用

目的研究肿瘤坏死因子α1型受体(tumor necrosis factor α receptor 1,TNFαR1)信号通路在小鼠急性后肢缺血中的作用,并探讨其作用机制.方法通过结扎TNFαR1-/-和野生型小鼠股动静脉及其主要分支建立后肢急性缺血模型,激光多普勒测手术前后后肢血液灌流,TUNEL染色观察细胞凋亡,凝胶电泳观察DNA凋亡梯带,免疫蛋白印迹法测定Caspase-3、Bax蛋白的表达.结果术后1天TNFαR1-/-组缺血侧后肢血液灌流显著高于野生型组,腓肠肌TUNEL阳性细胞显著低于野生型组.TNFαR1-/-组缺血评分则显著低于野生型组,两组的自发截肢率分别为50%、0%.TNFαR1-/-组DNA凋亡梯带、Caspase-3和Bax蛋白表达减弱.结论TNFαR1敲除可抑制TNF α信号通路下游死亡相关蛋白的激活,减少细胞死亡和调亡,对急性后肢缺血有保护作用.

Aim: To observe the apoptotic changes following exposure to EMP and to explore the possible injury mechanism in NIH - 3T3 fibroblasts. Methods: Following NIH - 3T3 cells were exposed to EMP,the proliferation and viability of NIH - 3T3 fibroblasts were estimated by hemacytometer and MTT Measurements. Apoptotic rate was measured by flow cytometer. The imnmohistochemical SP method was used to determine bcl- 2, p53. The positively stained cells were analyzed with CMIAS- Ⅱ image analysis system at a magnification 400. All data were analyzed by SPSS8.0 software. Results: The proliferation and viability of NIH- 3T3 cells were markedly inhibited and significant apoptosis was induced following exposure to EMP.EMP could increase the number of non- adherent cells, the highest ratio (33.9%) of non- adherent cells also occurred at 6h. The As70 value of MTT decreased immediately at 1 h, 6h following the cells were exposed as compared with the control (P ＜ 0.05). The highest ratio of apoptosis was 15.07% at 6h, then decreased gradually. Down - regulation of bcl - 2 and up - regulation of p53 were induced by EMP ( P ＜ 0.05). Conclusions: EMP could promote apoptosis of NIH - 3T3 fibroblasts. EMP could also down - regulate bcl - 2 level and up - regulate p53 level in NIH - 3T3 fibroblasts. Bcl - 2 and p53 gene may take part in the process of apoptosis.

AIM To assess the relationship between HBV X-gene, X-gene product and Fas/ FasL which mediatehepatocellular apoptosis in patients with hepatocellular carcinoma.METHODS Tissue from 34 patients with hepatocellular carcinoma was tested for the expression of HBxAg.Quantitative ELISA assay was used to detect sFas; and sFasL and PCR were used to detect the HBV X-genein sera from 30 patients with hepatocellular carcinoma, 32 patients with liver cirrhosis and 20 normalcontrols.RESULTS The positive expression of HBxAg, Fas and FasL in carcinoma tissue was 97.06%, 85.29% and100%, respectively. The positive signal was mainly presented in the plasma, and all of these three positivestaining may appear in the same area. Redit analysis showed that there was no significant difference amongthese three positive staining (P >0.05). The mean levels of sFas in sera from hepatocellular carcinoma, livercirrhosis and normal controls were 722.97±321.12, 801.90±419.94 and 224.07±148.23, respectively,showing that sFas levels in patients with hepatocellular carcinoma and liver cirrhosis were significantlyelevated than that in normal controls (P < 0.0l). The mean levels of sFasL in sera from hepatocellularcarcinoma, liver cirrhosis and normal controls were 152.27±7.99, 162.97±12.40 and 154.99 ± 6.96,showing that sFasL level in patients with liver cirrhosis was significantly higher than that in patients withhepatocellular carcinoma and normal controls (P< 0.01). HBV X-gene was found to be positive in sera of30% patients with hepatocellular carcinoma; HBV X-gene was found to be positive in sera of 43.75% ofpatients with liver cirrhosis. There was no significant difference in sFas/sFasL level between HBV X-genepositive patients and HBV X-gene negative patients (P >0.05).CONCLUSION The expression of HBxAg and Fas/FasL in the tissue of hepatocellular carcinoma seemed tobe almost the same, but relation between cause and effect is unclear. The detection of sFas and sFasL inpatient sera may reflect the state of apoptosis mediated by Fas/FasL system. Our data showed that HBV X-gene expression in sera seemed to have no relation to sFas/sFasL level; however, these data also suggestedthat some patients with negative HBsAg in sera might have integrated HBV X-gene in liver tissues, andtherefore X-gene is detectable in those patient sera.

AIM To study the effect of a wide range of concentration of arsenic trioxide on human hepatoma cell lineBEL-7402 and its mechanism.METHODS The BEL-7402 cells were treated with arsenic trioxide (a final concentration of 0.5, 1 and2 μmol/L, respectively) in various durations or for 4 successive days. The cell growth and proliferation wereobserved by cell counting and cell-growth curve. Morphologic changes were studied under electronmicroscopy. Flow cytometry was used to assay cell-DNA distribution and the protein expression of Bcl-2 andBax was detected by immunocytochemical method.RESULTS The cell growth was significantly inhibited by the different concentrations of arsenic trioxide asrevealed by cell counting and cell-growth curve. Arsenic trioxide treatment at 0.5, 1 and 2 μmol/L, resultedin a sub-G1 cell peak. The decreased G0/G1 phase cell and the increased percentage of S phase cell were observed by flow cytometer, suggesting that the inhibiting effect of arsernic trioxide on BEL-7402 cell lay inG0/G1 phase cell. Apoptotis-related morphology, such as intact cell membrane, nucleic condensation,apoptotic body formation, can be seen under the electron microscopy. High protein expression level of Bcl-2and Bax was detected in 1 and 2 μmol/L arsenic trioxide-treated cells, but that of Bax was more significant.Arsenic trioxide treatment at 0.5 μmol/L resulted in higher expression level of Bcl-2 and lower expressionlevel of Bax compared with control (P1<0.01, P2<0.01).CONCLUSION Arsenic trioxide not only inhibited the proliferation but also induced apoptosis of humanhepatoma cell line BEL-7402. The induced-apoptosis effect of 1 and 2 μmol/L arsenic trioxide was relative tothe expression level of Bcl-2 and Bax.

AIM To review the progress in pharmacological mechanisms of terandrine (Tet) and its therapeutic use indigestive diseases.METHODS We reviewed almost all the papers related to Tet from various magazines published in Englishand Chinese in recent years.RESULTS It has been demonstrated that Tet had multiple bioactivities: ① Tet could act as a Ca2+antagonist via blocking cellular plasma membrane voltage- or receptor-operating Ca2+ channels, inhibiting extracellular Ca2+ entry into the cell and intracellular Ca2+ mobilization to the cytosol, so as to preventhepatocytes, cardiomyocytes, pancreas cells and neurocytes from toxic or ischemia-reperfusion injuries.However, in HL-60 and leukemic T cells, Tet promoted Ca2+ releasing from mitochondria and microsomes,increased the concentration of intracellular Ca2 + , and induced cell death; ② Tet inhibited phobol 12-myristat13-acetate (PMA) plus ionomycin-induced T cell proliferation, interleukin-2 secretion and expression of theT cell activation antigen, CD71. It could also interrupt the integrity of macrophages, and reduced respiratoryburst of neutrophils and macrophages and proinflammatory cytokines secretion through minimizing nucleartranscriptional factor kappa B DNA binding activity; ③ Tet could induce tumor cell apoptosis, and down-regulate P-glucoprotein activity; and ④ Tet has the therapeutic effects on hepatic fibrogenesis, portalhypertension, immunomodulation, etc.CONCLUSION Tet can act as a Ca2 + channel blocker, inhibit proinflammatory factors releasing, modulateimmunoreaction, and induce tumor cell apoptosis. It can be used to prevent hepatocyte injury induced bytoxins and virus, inhibit hepatic fibrogenesis, reduce portal venous pressure, and can be used as an anti-tumor drug as well.

AIM To explore the therapeutic potential of antisense oligodeoxynucleotides on hepatcellular cacinoma(HCC).METHODS Four antisense phosphorothioated oligodeoxynucleotides (asON), complementary to differentsites of HBV, were synthesized and assayed for their anti-HBV activity in HepG22. 2.15 cells with ELISA.The most effective asON was chosen for the following study: FACSCAN, TRAP and immuno-staining wereused respectively for checking apoptosis, telomerase activity and expression of oncogene p21ras and p62C-myc inHepG2.2.15 cells after treated by asON.RESULTS The oligomer directed against the initiator of pre-S2 was the most effective one with aninhibitory rate of 66% on HBsAg and 91% on HBeAg (P<0.02). Two inhibitory peaks (bimodal)appeared. Telomerase activity as well as the expression of p21fas and p62C-myc decreased drastically 3 days afterasON-HBpreS2 treatment. Meanwhile, apoptosis appeared in the experiments.CONCLUSION The inhibitory effects of as-preS2 on the HBV gene expression and the reversion of somemalignant behaviour in HepG2.2.15 cells were the significant, effective therapy against HBV infection andhepatocellular carcinoma.

AIM To determine NO, NO synthase (NOS) and NOSmRNA of the esophageal carcinoma cells (SHEEC1)in apoptotic process induced by As2O3 and to explore the relationship between NO and apoptosis.METHODS The apoptosis of the cell line (SHEEC1) was induced by arsenite (As2O3, 5 μmol/L and10 μmol/L). In the process, at 2 h, 4 h, 8 h, 16 h and 24 h after administration of As2O3, NO production incultural medium was detected quantitatively by spectrophotometry; NOS Ⅱ was detected byimmunohistochemistry and NOS mRNA by in situ hybridization (ISH). The cells at endpoint of theexperiment were examined under transmitted electron microscope (TEM) for apoptosis.RESULTS The amount of NO released from SHEEC1 were increased from the basal condition (0.68×10-2μmol/L) up to the high level (2.38×10-2μmol/L) at h 16. The increment of NOS Ⅱ was found afteradministration of As2O3; the intracytoplasmic ISH signals of NOSmRNA in small size was found firstly at4 h, and then became highly predominant. Apoptotic changes of SHEEC1 occurred at 24 h under TEM.CONCLUSION After administration of As2O3, NO released from cultured SHEEC1 cells was detected withincreasing amount up to 16 h. The expression of NOS H and transcription of NOSmRNA are upregulated.The present findings suggest a concept that the NO may be a mediated and effective factor in apoptosisinduced by As2O3,

AIM To explore the anti-tumor effect of indomethacin (IN) on human colon adenocarcinoma cells anddetermine the influence of indomethacin on cancer cell proliferation and apoptosis and elucidate the anti-tumor mechanism of Indomethacin.METHODS Human colon adenocarcinoma HCT116 cell line were cultured separately in vitro. Indomethacin(final concentration 100 μm - 800 gm) was administered alone or altogether with 5-Fu (50 μm). Agarose gelelectrophoresis, MTT, and Flow cytometry were used to study cell proliferation and apoptosis in humancolon carcinoma cell RT-PCR, western blot were used to detect the expression level of Bcl-2, bax gene andcdk4 protein expression in HCT116 cell lines after treated with IN for 24 hours.RESULTS Indomethacin can inhibit significantly the proliferation of HCT116 cell, change the morphology,and cause the cells to accumulate in the G0/Gl phase of the cell cycle, and induce apoptosis. The apoptosis oftumor cells was confirmed by DNA ladder formation on gel electrophoresis and sub-Gl peak on flowcytometry. These responses were time-and concentration-dependent. A synergic effect of inhibiting cancercell proliferation was observed when combined with Indomethacin and 5-Fu. RT-PCR results showed that INdown-regulated Bcl-2 mRNA expression, and did not change Bax mRNA expression. Western blot resultsconfirmed that IN inhibited Bcl-2 protein expression. No influence was found in the translation of Baxprotein. In inhibited cdk4 protein expression.CONCLUSION Our study results indicate that IN induce apoptosis of HCT116 cell by down-regulating Bcl-2expression and inhibiting cdk4 protein expression partially. This explains the mechanisms of antitumoractivity of the Indomethacin.

AIM To study the combined expression of gastrointestinal hormone substance P and anti-apoptosis gene Bcl-2 in gastric carcinoma and its significance.METHODS Substance P and Bcl-2 protein expression was examined by the S-P immunohistochemicalmethod in 33 cases of gastric carcinoma, 17 adjacent the carcinoma and 13 normal gastric mucoma.RESULTS Positive expression of SP in gastric carcinoma was higher than that of both adjacent and normalmucosa (P < 0.001). There was no statistical difference in the positive expression between adjacent andnormal mucosa (P > 0.05). The expression of bcl-2 both in gastric carcinoma and adjacent tissues werehigher than that of normal gastric mucosa (P< 0.05-0.01). But the positive expression of Bcl-2 had nostatistical significance between gastric carcinoma and adjacent tissues.CONCLUSION Both gastrointestinal hormone SP and Bcl-2 gene have synergistic expression in gastriccarcinoma, indicating that they all take part in the occurrence of gastric carcinoma. Abnormal expression ofBcl-2 gene occurred in benign gastric pathological changes, once they become carcinoma, the positiveexpression of cell is no more increased, possibly because that there is no more increase of the intensity of Bcl-2 inhibition of cell apoptosis.

AIM To understand the rule and possible function of apoptosis and protein expression of bcl-2, p53 and C-myc in chronic gastritis, gastric ulcer, non-classic proliferation of gastric mucosa and gastric cancer.METHODS Apoptosis was detected by using in situ terminal labelling (TUNEL). The protein expression ofbcl-2, p53 and C-myc was detected by immunohistochemical method.RESULTS The indexes of apoptosis in chronic active gastritis, gastric ulcer, mild and severe non-classicproliferation of gastric mucosa, early and progressive gastric cancer were 16.8%±12.3%, 24.1%±20.0%,19.3%±16.4%, 15.7%±15.2%, 10.1%±9.1% and 6.3%±6.0%, respectively. The index of progressivegastric cancer was lower than that of early gastric cancer and non-classic proliferation of gastric mucosa(P<0.05). The positive rate of bcl-2 protein was 9.4%, 27.6%, 52.9%, 75.0%, 83.3% and 46.7%,respectively. The positive rate of bcl-2 of early gastric cancer was higher than that of progressive gastriccancer. The positive rates of p53 protein of severe non-classic proliferation, early and progressive gastriccancer were 25.0%, 33.3% and 63.3%, respectively. The positive rate of p53 of progressive gastric cancerwas higher than that of early gastric cancer and non-classic proliferation (P<0.05). In Lauren types, theindex of apoptosis, protein expression rates of bcl-2, p53 and C-myc of intestinal type were 8.3%±7.2%,38.9%, 77.7% and 56.6%, while that of diffuse type were 5.1%±4.9%, 58.3%, 50.0% and 8.3%,respectively. All markers had statistical difference between two types (P<0.05).CONCLUSION Apoptosis was inhibited stepwise in the development of non-classic proliferation of gastricmucosa to early gastric cancer and then to progressive gastric cancer. The high expression of bcl-2, p53 andC-myc was related to the development of gastric cancer, bcl-2 might play an important role in early gastriccancer while p53 and C-myc act mostly in middle and late stage gastric cancer. The Lauren typing of gastriccancer is closely related to the index of apoptosis and expression of bcl-2, p53 and C-myc.

INTRODUCTIONThe treatment of human epithelial malignancies is limited by drug resistance and toxic and side effects,which results in the failure in the treatment of majority of advanced cancer victims. To seek for a new, and specific antineoplastic therapy will provide hope for tumor treatment. Although disordered intermediary metabolism in cancer cells has been known for many years, much of the work focused on abnormal glucose catabolism. At the same time, little attention has been paid to fatty acid synthasis in tumor tissues, dispite of the significance of fatty acid synthase (FAS) in some clinical human ovarian[1], breast[2], colorectal[3],and prostatic cancers[4,5]. Tumor cells which express high levels of fatty acid synthesizing enzymes use endogeneously synthesized fatty acids for membrance biosynthesis and appear to export large amounts of lipid. In contrast, normal cells preferentially utilize diary lipid.

INTRODUCTION The antitumor activity of norcantharidin (NCTD),the demethylated analogue of cantharidin, was studied in the early 1980s in China. NCTD has no side effects on urinary organs which cantharidin has shown and is easier to synthesize, and it can inhibit the proliferation of several tumor cell lines as well as transplanted tumors. Clinical trials with NCTD as a monotherapeutic agent indicated that NCTD had beneficial effects in patients with different kinds of digestive tract cancers, such as primary hepatoma,carcinomas of esophagus and gastric cancer, but no depressive effect on bone marrow cells. NCTD can increase the white blood cell count by stimulating the bone marrow and has some antagonistic effect against leukopenia caused by other agents. The exact cellular and molecular mechanisms of NCTD on tumor cells have not yet been elucidated to date[1-3].