Anisomycin诱导小鼠艾氏腹水癌细胞凋亡

目的：探讨Anisomycin诱导EAC细胞凋亡的机制。方法：利用MTT法观察Anisomycin对EAC细胞增殖的影响;应用An．nexinV—FITC和PI双染色检测Anisomycin作用下EAC细胞凋亡的变化;琼脂糖凝胶电泳检测Anisomycin作用下EAC细胞的DNA断裂片断;采用RT—PCR检测Anisomycin作用下Caspase-3mRNA的转录水平;WesternBlot分析Anisomycin处理的EAC细胞内Caspase-3蛋白的表达。结果：Anisomycin对EAC细胞增殖的抑制率随浓度的升高而增高,EAC细胞的凋亡水平也上升,随着Anisomycin浓度升高EAC细胞DNA断裂的寡核苷酸片段亦越趋明显,Caspase-3的mRNA和蛋白水平也上调,并明显高于阿霉素组。结论：结果表明,Anisomycin能够抑制EAC细胞的增殖,在体外可能通过激活Caspase-3凋亡信号而诱导EAC细胞的凋亡。     

Anisomycin诱导小鼠艾氏腹水癌细胞凋亡

目的:探讨Anisomycin诱导EAC细胞凋亡的机制。方法:利用MTT法观察Anisomycin对EAC细胞增殖的影响;应用AnnexinV-FITC和PI双染色检测Anisomycin作用下EAC细胞凋亡的变化;琼脂糖凝胶电泳检测Anisomycin作用下EAC细胞的DNA断裂片断;采用RT-PCR检测Anisomycin作用下Caspase-3 mRNA的转录水平;Western Blot分析Anisomycin处理的EAC细胞内Caspase-3蛋白的表达。结果:Anisomycin对EAC细胞增殖的抑制率随浓度的升高而增高,EAC细胞的凋亡水平也上升,随着Anisomycin浓度升高EAC细胞DNA断裂的寡核苷酸片段亦越趋明显,Caspase-3的mRNA和蛋白水平也上调,并明显高于阿霉素组结论:结果表明,Anisomycin能够抑制EAC细胞的增殖,在体外可能通过激活Caspase-3凋亡信号而诱导EAC细胞的凋亡。     

不同单位保种的艾氏腹水癌细胞核型及DNA含量的比较研究

目的 比较研究三个单位保种的艾氏腹水癌细胞（EAC）染色体数目及DNA含量。方法 取腹腔接种EAC 6 d的KM小鼠,腹腔注射秋水仙碱,取腹水,经低渗处理,固定后涂片,染色,显微镜下染色体计数;腹水传代的EAC,用70%乙醇固定,流式细胞仪测DNA含量。结果 山东省医学科学院药物研究所（山东）、武汉大学典型培养物保藏中心（武汉）和北京市肿瘤研究所（北京）保种的艾氏腹水癌细胞株,其染色体均数分别为28.31±7.78、29.16±12.25、23.22±10.21条,方差分析表明,除北京与山东外,武汉与北京、武汉与山东两两比较有极显著差异（P<0.01）。直方图分析显示主流染色体范围分别为25-27、22-24、22-24条。北京艾氏腹水癌细胞 DNA含量最多,山东艾氏腹水癌细胞DNA含量最少。结论 三个单位的艾氏腹水癌细胞株在某些方面已出现差异。     

艾氏腹水癌患鼠腹水中一种高分子量DNA结合蛋白的分离

 本文利用免疫吸收法和免疫亲和层析法,从艾氏腹水癌患鼠腹水DNA结合蛋白中,分离得到了一种高分子量DNA结合蛋白。在免疫双扩散反应中,它与抗艾氏腹水癌患鼠血清DNA结合蛋白的兎抗血清反应形成一条沉淀线,但与正常小鼠血清DNA结合蛋白的兎抗血清不形成沉淀线。该DNA结合蛋白样品用2-巯基乙醇还原后,经SDC-PAGE分析,测得其分子量约为41000。     

三单位保存的艾氏腹水癌细胞株及克隆细胞蛋白质表达

目的　比较三个单位保存的艾氏腹水癌 (EAC)细胞株及克隆细胞的蛋白质表达。方法　对北京市肿瘤研究所 (北京 )保存的EAC进行克隆培养 ,从中选出 5株克隆 ;对武汉大学保种中心 (武汉 )、山东省医学科学院药物研究所 (山东 )及北京保存的EAC细胞株及 5株克隆细胞的SDS PAGE电泳图谱及免疫组化蛋白质分布进行了对比。结果　武汉、山东及北京保存的EAC的电泳条带数分别为 2 2、2 5及 2 8条 ,而克隆细胞E2G8为 2 6条带。三单位EAC瘤细胞对 5种抗体做免疫组化染色 ,与 1株抗体反应的阳性细胞比例均较高 ,差异无显著性 (P >0 0 5 ) ,与另 4种抗体反应的阳性细胞率差异有显著性 ;5株克隆细胞对 10种抗体作免疫组化染色 ,其中克隆细胞E2G8、E2F4与 7种抗体反应阳性 ,E2C6与 8种抗体反应阳性 ,E1G5、E2B5与 6种抗体反应阳性。克隆细胞一旦与某种抗体反应阳性 ,阳性细胞的比例即在 85 %以上。结论　不同单位保存的EAC细胞株及克隆细胞株蛋白质表达有差异     

Chemical Constituents of Cape Aloe and Their Synergistic Growth-Inhibiting Effect on Ehrlich Ascites Tumor Cells

The constituents of cape aloe were investigated after a preliminary screening of the growth-inhibiting effect on Ehrlich ascites tumor cells (EATC) of several extracts of this plant. Ten compounds were isolated from the dichloromethane (CH₂Cl₂) extract that showed the strongest activity, and their structures were elucidated as aloe-emodin (1), p-hydroxybenzaldehyde (2), p-hydroxyacetophenone (3), pyrocatechol (4), 10-oxooctadecanoic acid (5), 10-hydroxyoctadecanoic acid (6), methyl 10-hydroxyoctadecanoate (7), 7-hydroxy-2,5-dimethylchromone (8), furoaloesone (9), and 2-acetonyl-8-(2-furoylmethyl)-7-hydroxy-5-methylchromone (10) based on MS and various NMR spectroscopic techniques. Compounds 2–7 were isolated for the first time from cape aloe. Compounds 4–7 and 10 showed a significant growth-inhibiting effect, and compound 1 exhibited a remarkable synergistic effect on compounds 8–10, which was not observed with the treatment by each compound alone on EATC. These results suggest that the strong growth-inhibiting effect of the CH₂Cl₂ extract was dependent not on one compound alone, but on the synergistic effect from the combination of compound 1 and the other compounds.     

Characterization of gangliosides from Ehrlich ascites tumour cells and their variants

Differences in the nature of the gangliosides present in two types of Ehrlich ascites tumour (EAT) cells, the adherent and non-adherent EAT cells, were studied. Gangliosides were isolated by DEAE Sephadex column chromatography and analysed by high-performance thin-layer chromatography (HPTLC). The non-adherent EAT (na-EAT) cells which grow in the peritoneal cavity of mice were selected for growth on basement membrane and tissue culture plastic to give the adherent EAT (a-EAT) cells. na-EAT cells contained 1.57 nmol lipid-bound sialic acid per mg protein and at least 12 different gangliosides, including major gangliosides such as GM3, GM2, GM1, GD3, GD1a and GT1b. On the other hand, the ganglioside pattern of a-EAT cells differed significantly from that of na-EAT cells, both quantitatively and qualitatively. The content of lipid-bound sialic acid in a-EAT cells was only 0.24 nmol per mg of protein. The gangliosides in a-EAT cells were characterized as GD1a and trisialogangliosides and, significantly, a-EAT cells did not contain monosialogangliosides. Neutral glycolipids were isolated from both cell lines and their patterns were compared. In contrast to the gangliosides pattern, their neutral glycolipid patterns were similar. Glucosylceramide and lactosylceramide were the major components in both types of cells. In addition to na- and a-EAT cells, a-EAT cells were passaged in mice by intraperitoneal injection, giving rise to a third variant (c/m EAT cells). We analysed the gangliosides in c/m EAT cells to determine whether there was a change in the ganglioside pattern found in na-EAT cells. After repeated passage of c/m EAT cells in mice, the pattern of gangliosides shifted to that of na-EAT cells. Alterations of ganglioside composition may be associated with the growth environment of the murine peritoneal cavity; alternatively, a selection process may have occurred.Abbreviations EAT cells Ehrlich ascites tumour cells - na-EAT cells non-adherent EAT cells - a-EAT cells adherent EAT cells - c/m EAT cells cultured a-EAT cells passaged in mice - HPTLC high-performance thin-layer chromatography - PBS 10 mM phosphate-buffered saline, pH 7.2, containing 0.15 M NaCl - EDTA ethylene-diaminetetraacetic acid - TFA trifluoroacetic acid - TG thioglycollate - Cer ceramide (N-fatty acyl sphingosine) - GM3 NeuAc2-3Gal1-4Glc-Cer - GM2 GalNAc1-4(NeuAc2-3)Gal1-4Glc-Cer - GM1a Gal1-3GalNAc1-4(NeuAc2-3)Gal1-4Glc-Cer - GD3 NeuAc2-8NeuAc2-3Gal1-4Glc-Cer - GD1a NeuAc2-3Gal1-3GalNAc1-4(NeuAc2-3)Gal1-4Glc-Cer - GT1b NeuAc2-3Gal1-3GalNAc1-4(NeuAc2-8NeuAc2-3)Gal1-4Glc-Cer - LacCer Gal1-4Glc-Cer - Gb3 Gal1-4Gal1-4Glc-Cer - Gb4 GalNAc1-4Gal1-4Gal1-4Glc-Cer This paper is dedicated to my esteemed colleague, Sen-itiroh Hakomori on the occasion of his 65th birthday.     

Subcutaneous Growth of Staphylococcus aureus Concomitantly Inoculated with Ehrlich Ascites Tumor Cells

Intratumoral growth of Staphylococcus aureus Cowan I-derived AP332 was examined by subcutaneous inoculation of cocci in doses ranging from 18 to 1.8 × 10⁵ CFU with Ehrlich ascites tumor cells. Inoculation of 18 CFU AP332 resulted in staphylococcal growth in one of five mice, and the proportion of mice established intratumoral infection increased with the initial inocula. Six other strains of S. aureus also grew in the tumor tissue, and none of the three strains of coagulase-negative staphylococci grew at all. Ethanol-killed tumor cells did not promote staphylococcal growth as vigorously as the live tumor cells, especially when the initial inoculum of AP332 was smaller than 10⁴ CFU.     

Effect of weak combined static and extremely low‐frequency alternating magnetic fields on tumor growth in mice inoculated with the Ehrlich ascites carcinoma

It has been shown that the ultralow‐frequency extremely weak alternating component of combined magnetic fields (MFs) exhibits a marked antitumor activity. The parameters of this component have been found (frequency 1, 4.4, 16.5 Hz or the sum of these frequencies; intensity 300, 100, 150–300 nT, respectively) at which this MF in combination with a collinear static MF of 42 µT inhibits or suppresses the growth of Ehrlich ascites carcinoma (EAC) in mice. It was shown that the exposure of mice with EAC to combined MFs causes structural changes in some organs (liver, adrenal glands), which are probably due to the total degradation of the tumor tissue. In mice with transplanted EAC, the tumor tissue after exposure to weak MFs was practically absent, as distinct from control animals in which the invasion of the tumor into the adipose tissue surrounding the kidneys, mesenteric lymph nodes, and spermatic appendages was observed. In animals without tumors, no pathological deviations from the norm in the structure of organs and tissues occurred after exposure to weak MF, indicating that this factor per se is not toxic to the organism. Bioelectromagnetics 30:343–351, 2009. © 2009 Wiley‐Liss, Inc.     

Isolation and partial characterization of the ADP-ribosylated nuclear proteins from Ehrlich ascites tumor cells

The (ADP-ribose)_n protein conjugates formed by incubation of Ehrlich ascites tumor cell nuclei with 1 mM (³H)NAD were isolated by chromatography on boronate cellulose columns with a yield of >85%. Possible contamination by glycoproteins was excluded by rechromatography after specific release of the (ADP-ribose)_n residues from their acceptors. Dodecyl sulfate gel electrophoresis revealed numerous protein bands which coincided with the (³H)ADP-ribose bands obtained by fluorography of the gels. 40% of the acceptor proteins were identified as the nucleosomal core histones. Most of these histones, however, appeared in the non-histone fraction because of extensive modification by poly(ADP-ribose). Drastic changes in properties were also seen in the true non-histone proteins which comprised 60% of the total conjugated protein. Besides several prominent acceptor proteins (M_r = 12,000; 31,000; 125,000) numerous proteins were detected indicating a considerable heterogeneity of non-histone acceptors.     

Combination of arsenic trioxide and cisplatin synergistically inhibits both hexokinase activity and viability of Ehrlich ascites carcinoma cells

Hexokinase‐2 is overexpressed in several carcinomas including breast cancer to sustain energy for rapidly dividing cells and associates with chemoresistance. However, the impact of chemo drugs (alone or in combination) on hexokinase activity and autophagic cell death is unclear. In this report, we used an in vivo murine adenocarcinoma model to validate the effects of As₂O₃ and cisplatin on hexokinase activity and autophagic cancer cell death. We found that the two drugs inhibit hexokinase activity and induce autophagic marker, beclin 1 expression. Interestingly, combining As₂O₃ with cisplatin synergistically enhanced these effects and alleviated oxidative stress often encountered in As₂O₃ treatment. Altogether, our data provide direct evidence that inhibition of hexokinase activity and induction of autophagic cell death are mediating the antineoplastic effects of As₂O₃ and cisplatin. Our findings raise the potential of combining As₂O₃ with cisplatin as an approach to augment cisplatin‐induced cell death and combat cisplatin chemoresistance in cancer.     

Antioxidant effect of zinc and zinc-metallothionein in the acute cytotoxicity of hydrogen peroxide in Ehrlich ascites tumour cells

This study was concerned with the role of zinc (Zn) and zinc-metallothionein (Zn-MT) in oxidative stress. Hydrogen peroxide-induced oxidative injury was examined in Ehrlich ascites tumour cells isolated from control host mice, mice pretreated with 10 mg/kg ZnSO4 (i.p.) to increase cellular Zn/Zn-MT levels, and mice exposed to Zn-deficient diet to reduce the cellular Zn/Zn-MT levels. The results of the present study showed that Ehrlich cells with seven-fold differences in Zn-MT concentrations could be obtained by manipulating the Zn status of host mice and that high Zn and Zn-MT levels can make Ehrlich cells more resistant to H2O2-induced oxidative injury (cell viability, lipid peroxidation, [Ca2+]i) while cells with reduced Zn/Zn-MT levels were more susceptible to this treatment. H2O2 treatment resulted in oxidation of MT thiolate groups and loss of its metal binding capacity with translocation of Zn released from oxidized MT to other cellular sites. Preincubation of Ehrlich cells with ZnSO4 in vitro also conferred some degree of resistance to H2O2 toxicity, suggesting the inherent antioxidative property of Zn ions. These data suggested that Zn-MT can be considered as an antioxidant by virtue of its thiolate groups and its Zn ions that are released in the presence of oxidative stress.     

Involvement of protein tyrosine phosphorylation and reduction of cellular sulfhydryl groups in cell death induced by 1' -acetoxychavicol acetate in Ehrlich ascites tumor cells

Elucidation of the mechanisms underlying potential anticancer drugs continues and unraveling these mechanisms would not only provide a conceptual framework for drug design but also promote use of natural products for chemotherapy. To further evaluate the efficacy of the anticancer activity of 1'-acetoxychavicol acetate (ACA), this study investigates the underlying mechanisms by which ACA induces death of Ehrlich ascites tumor cells. ACA treatment induced loss of cell viability, and Western blotting analysis revealed that the compound stimulated tyrosine phosphorylation of several proteins with 27 and 70 kDa proteins being regulated in both dose- and time-dependent manner prior to loss of viability. Protein tyrosine kinase inhibitor herbimycin A moderately protected cells from ACA-induced toxicity. In addition, cellular glutathione and protein sulfydryl groups were also significantly reduced both dose- and time-dependently during evidence of cell death. Replenishing thiol levels by antioxidant, N-acetylcysteine (NAC), an excellent supplier of glutathione and precursor of glutathione, substantially recovered the viability loss, but the recovery being time-dependent, as late addition of NAC (at least 30 min after ACA addition to cultures) was, however, ineffective. Addition of NAC to ACA treated cultures also abolished tyrosine phosphorylation of the 27 kDa protein. These results, at least partly, identify cellular sulfhydryl groups and protein tyrosine phosphorylation as targets of ACA cytotoxicity in tumor cells.     

Exacerbation of Experimental Trypanosoma cruzi Infection in Mice by Concomitant Malaria*

SYNOPSIS. The effect of malaria on the chronic phase of Chagas’disease was investigated in mice. The animals were given Plasmodium berghei-infected red blood cells 2 to 12 months after their initial inoculation with trypomastigotes of 3 different strains of Trypanosoma cruzi (Y, CL and Gilmar). In all the experiments carried out with one of the strains (CL), a somewhat variable but always considerable percentage of mice (average 39%) relapsed in to the acute phase of Chagas’disease. This relapse was characterized by a significant increase in the number of circulating trypomastigotes. Recrudescence was observed also with a 2nd strain of T. cruzi (Gilmar), which is similar in many aspects to the CL strain, e.g. the morphology of blood stages, curve of parasitemia and susceptibility to antibodies in vitro. In mice whose chronic phase was induced by trypomastigotes of the Y strain, malaria infections did not induce a typical acute phase with high parasitemia by T. cruzi. Bloodstream forms of Y parasites differ from those of CL and Gilmar strains morphologically as well as immunologically, i.e. only the Y strain is easily agglutinated and partly inactivated by specific immune serum. In light of this and other known characteristics of the strains used in the present work, the author speculates on mechanisms which allow malaria infections selectively to suppress acquired host resistance to certain strains of T. cruzi.     

不同FIGO分期子宫内膜癌患者外周血红细胞分布宽度、血清对氧磷酶-1表达水平差异及其诊断价值分析

摘要 目的：探讨不同国际妇产科学联合会（FIGO）分期子宫内膜癌患者外周血红细胞分布宽度（RDW）、血清对氧磷酶-1（PON-1）表达水平差异及其诊断价值。方法：选取我院2018年1月到2020年1月收治的80例子宫内膜癌患者作为研究对象,根据FIGO分期对所有患者进行分组,分为Ⅰ期组23例,Ⅱ期组22例,Ⅲ期组18例,Ⅳ期组17例。另选取同期来我院体检的30名健康志愿者作为对照组,对比所有受检者RDW、PON-1表达水平,并应用ROC曲线分析外周血红细胞分布宽度、血清对氧磷酶-1表达水平对不同FIGO分期子宫内膜癌的诊断价值。对80例子宫内膜癌患者进行2年随访,将2年内死亡的25例患者分为死亡组,将其余55例患者分为存活组,对比死亡组与存活组临床一般情况与RDW、PON-1表达水平,并应用logistic回归分析分析RDW、PON-1对子宫内膜癌的预后预测价值。结果：五组受检者RDW、PON-1水平对比差异显著,Ⅳ期组RDW水平高于Ⅲ期组、Ⅱ期组、Ⅰ期组和对照组,Ⅳ期组PON-1水平低于Ⅲ期组、Ⅱ期组、Ⅰ期组和对照组（P＜0.05）;RDW结合PON-1联合诊断的准确度、敏感度、阳性预测值高于RDW单一诊断和PON-1单一诊断（P＜0.05）。RDW诊断子宫内膜癌不同分期价值曲线下面积为89.63,最佳诊断着色界限值为75.73 %,PON-1的曲线下面积为78.89,最佳诊断着色界限值为82.53 %,两者联合的曲线下面积为84.26,最佳诊断着色界限值为87.57 %;存活组与死亡组患者性别、年龄、病理类型对比无明显差异（P＞0.05）,两组患者FIGO分期、基层浸润、组织分化程度、血清RDW与PON-1表达水平对比差异显著（P＜0.05）;logistic回归分析结果表明：基层浸润、RDW与PON-1为子宫内膜癌患者预后的独立影响因素（P＜0.05）。结论：RDW、PON-1联合对子宫内膜癌FIGO分期的诊断价值较高,且基层浸润、RDW与PON-1为子宫内膜癌患者预后的独立影响因素。临床上需针对基层浸润、RDW升高与PON-1水平降低的患者采取相关措施,改善治疗方案,降低患者死亡率情况。     

Transient generation of hydrogen peroxide is responsible for carcinostatic effects of hydrogen combined with platinum nanocolloid,together with increases intracellular ROS,DNA cleavages,and proportion of G2/M-phase

In our previous study, we demonstrated that combined treatment with hydrogen (H₂) and platinum nanocolloid (Pt-nc) exerted markedly antiproliferative effects on cancer cells compared with each treatment alone. However, because the related mechanisms remain unclear, we investigated carcinostatic mechanisms of the combined treatment with H₂ + Pt-nc. Significant suppression of cell proliferation was confirmed at 52?h following combined treatment, and the similar effect was also observed by the 30- or 40-min transient treatment with H₂?+?Pt-nc. The transient treatments led to changes in cell size and morphology, loss of microvilli, and apoptosis-like cell death at 120 h after treatment. Moreover, transient combined treatment with H₂?+?Pt-nc induced cell-cycle arrest, as reflected by decreased proportions of G1-phase cells and accumulation of G2/M-phase cells. In contrast, intracellular peroxide levels were temporarily and significantly increased immediately after H₂?+?Pt-nc treatment but not after treatment with H₂ or Pt-nc alone. Additionally, combined treatment-induced carcinostatic effects were significantly diminished in the presence of catalase, and marked hydrogen peroxide (H₂O₂) generation was confirmed after mixing Pt-nc into cell culture media containing a high concentration of H₂. These changes are in agreement with the results that carcinostatic effects were induced after only 40 min of treatment with H₂?+?Pt-nc. Thus, transient and marked generation of H₂O₂ is responsible for the carcinostatic effects of combined treatment with H₂?+?Pt-nc.     

Measuring Bacterial Load and Immune Responses in Mice Infected with Listeria monocytogenes

Listeria monocytogenes (Listeria) is a Gram-positive facultative intracellular pathogen¹. Mouse studies typically employ intravenous injection of Listeria, which results in systemic infection². After injection, Listeria quickly disseminates to the spleen and liver due to uptake by CD8α⁺ dendritic cells and Kupffer cells^3,4. Once phagocytosed, various bacterial proteins enable Listeria to escape the phagosome, survive within the cytosol, and infect neighboring cells⁵. During the first three days of infection, different innate immune cells (e.g. monocytes, neutrophils, NK cells, dendritic cells) mediate bactericidal mechanisms that minimize Listeria proliferation. CD8⁺ T cells are subsequently recruited and responsible for the eventual clearance of Listeria from the host, typically within 10 days of infection⁶.Successful clearance of Listeria from infected mice depends on the appropriate onset of host immune responses⁶ . There is a broad range of sensitivities amongst inbred mouse strains^7,8. Generally, mice with increased susceptibility to Listeria infection are less able to control bacterial proliferation, demonstrating increased bacterial load and/or delayed clearance compared to resistant mice. Genetic studies, including linkage analyses and knockout mouse strains, have identified various genes for which sequence variation affects host responses to Listeria infection^6,8-14. Determination and comparison of infection kinetics between different mouse strains is therefore an important method for identifying host genetic factors that contribute to immune responses against Listeria. Comparison of host responses to different Listeria strains is also an effective way to identify bacterial virulence factors that may serve as potential targets for antibiotic therapy or vaccine design.We describe here a straightforward method for measuring bacterial load (colony forming units [CFU] per tissue) and preparing single-cell suspensions of the liver and spleen for FACS analysis of immune responses in Listeria-infected mice. This method is particularly useful for initial characterization of Listeria infection in novel mouse strains, as well as comparison of immune responses between different mouse strains infected with Listeria. We use the Listeria monocytogenes EGD strain¹⁵ that, when cultured on blood agar, exhibits a characteristic halo zone around each colony due to β-hemolysis¹ (Figure 1). Bacterial load and immune responses can be determined at any time-point after infection by culturing tissue homogenate on blood agar plates and preparing tissue cell suspensions for FACS analysis using the protocols described below. We would note that individuals who are immunocompromised or pregnant should not handle Listeria, and the relevant institutional biosafety committee and animal facility management should be consulted before work commences.     

Revealing the Effects of the Herbal Pair of Euphorbia kansui and Glycyrrhiza on Hepatocellular Carcinoma Ascites with Integrating Network Target Analysis and Experimental Validation

Although the herbal pair of Euphorbia kansui (GS) and Glycyrrhiza (GC) is one of the so-called "eighteen antagonistic medicaments" in Chinese medicinal literature, it is prescribed in a classic Traditional Chinese Medicine (TCM) formula Gansui-Banxia-Tang for cancerous ascites, suggesting that GS and GC may exhibit synergistic or antagonistic effects in different combination designs. Here, we modeled the effects of GS/GC combination with a target interaction network and clarified the associations between the network topologies involving the drug targets and the drug combination effects. Moreover, the "edge-betweenness" values, which is defined as the frequency with which edges are placed on the shortest paths between all pairs of modules in network, were calculated, and the ADRB1-PIK3CG interaction exhibited the greatest edge-betweenness value, suggesting its crucial role in connecting the other edges in the network. Because ADRB1 and PIK3CG were putative targets of GS and GC, respectively, and both had functional interactions with AVPR2 approved as known therapeutic target for ascites, we proposed that the ADRB1-PIK3CG-AVPR2 signal axis might be involved in the effects of the GS-GC combination on ascites. This proposal was further experimentally validated in a H22 hepatocellular carcinoma (HCC) ascites model. Collectively, this systems-level investigation integrated drug target prediction and network analysis to reveal the combination principles of the herbal pair of GS and GC. Experimental validation in an in vivo system provided convincing evidence that different combination designs of GS and GC might result in synergistic or antagonistic effects on HCC ascites that might be partially related to their regulation of the ADRB1-PIK3CG-AVPR2 signal axis.     

DC-CIK治疗对晚期肝细胞癌患者免疫功能及循环肿瘤细胞的影响

目的:研究DC-CIK细胞治疗对晚期肝细胞癌患者甲胎蛋白、免疫功能及循环肿瘤细胞数的影响。方法:选取2013年至2015年我院收治的26例肝细胞癌并伴有复发或转移的患者,对其治疗前后甲胎蛋白、免疫功能及循环肿瘤细胞数的数据进行分析,比较DC-CIK细胞治疗前后甲胎蛋白、循环肿瘤细胞、免疫功能的变化。根据细胞治疗次数分为1次组及1次组,比较两组免疫功能的变化。结果:DC-CIK细胞治疗前甲胎蛋白为(603.32±155.78)ng/mL细胞回输后为(571.24±147.49)ng/mL差异无统计学意义(P0.05)。DC-CIK细胞治疗前及细胞回输后CTC检测阳性率分别为81.8%、36.4%治疗前及回输后循环肿瘤细胞数量分别为(8.36±10.642)、(1.55±2.464),细胞治疗前后比较差异均有统计学意义(P0.05)。细胞治疗前1天T细胞亚群CD3+、CD3~+CD4~+、CD3~+CD8~+、CD4/CD8分别为(66.05±15.31)%、(41.89±12.33)%、(23.15±8.05)%、(2.10±0.77),回输后分别为(69.69±12.91)%、(44.80±11.11)%、(23.13±7.12)%、(2.29±0.91)治疗前后比较差异无统计学意义(P0.05)。细胞治疗次数1次组和1次组免疫功能变化差异无统计学意义(P0.05)。结论:DC-CIK细胞治疗可减少肝细胞癌伴复发或转移患者循环肿瘤细胞的数量,但对甲胎蛋白和免疫功能无明显影响。