Cytotoxic effect of methanolic extracts of Fritillaria imperialis bulbs and Eryngium caucasicum leaves on hepatoma and colon cancer cells

Objective: To evaluate antitumor activities of Fritillaria imperialis and Eryngium caucasicum methanolic extracts on human hepatoma (HepG2) and colon cancer (HCT116) cell lines in comparison to human foreskin fibroblasts as the normal cells. Methods: Methanolic extracts of Fritillaria imperialis and Eryngium caucasicum were prepared by the maceration method. The effect of the extracts at various concentrations (100, 200, 400, 600, and 800 μg/mL) on cell survival was evaluated using the MTT method. Besides, fluorescence staining was used to evaluate death patterns of the cells. Results: MTT assay showed that Fritillaria imperialis significantly decreased the viability of all cell lines after 24 and 48 hours of treatments. However, Eryngium caucasicum extract did not show any significant cytotoxicity effect on the cell lines. Fluorescence staining revealed that Fritillaria imperialis induced apoptosis of HCT116 cells at 550 μg/mL. Conclusions: Fritillaria imperialis extract has antiproliferative and cytotoxic effects on HCT116 and HepG2 cancer cells and therefore, may serve as an anticancer agent.

Cytotoxic effect of methanolic extracts of Fritillaria imperialis bulbs and Eryngium caucasicum leaves on hepatoma and colon cancer cells

Objective: To evaluate antitumor activities of Fritillaria imperialis and Eryngium caucasicum methanolic extracts on human hepatoma (HepG2) and colon cancer (HCT116) cell lines in comparison to human foreskin fibroblasts as the normal cells. Methods: Methanolic extracts of Fritillaria imperialis and Eryngium caucasicum were prepared by the maceration method. The effect of the extracts at various concentrations (100, 200, 400, 600, and 800 μg/mL) on cell survival was evaluated using the MTT method. Besides, fluorescence staining was used to evaluate death patterns of the cells. Results: MTT assay showed that Fritillaria imperialis significantly decreased the viability of all cell lines after 24 and 48 hours of treatments. However, Eryngium caucasicum extract did not show any significant cytotoxicity effect on the cell lines. Fluorescence staining revealed that Fritillaria imperialis induced apoptosis of HCT116 cells at 550 μg/mL. Conclusions: Fritillaria imperialis extract has antiproliferative and cytotoxic effects on HCT116 and HepG2 cancer cells and therefore, may serve as an anticancer agent.

Construction of anti-BCMA chimeric antigen receptor (CAR-BCMA) modified T cells and its cytotoxicity against tumor cells / 中国肿瘤生物治疗杂志

@# Objective： ：To explore a novel chimeric antigen receptor (CAR)-T cell treatment to treat Multiple Myeloma (MM) via target B cell maturation antigen (BCMA). Methods： ：A CAR-BCMA molecular was constructed based on mouse originated BCMA scFv, and was packaged into lentiviral vector and transfected into T cells from healthy donors to construct CAR-BCMA-T cells. The BCMApositive cell lines A549-BCMA, A549-BCMAOFP and K562-BCMA were constructed as target cells. Then, the CAR-BCMA-T cells were co-incubated with the constructed target cells and human myeloma U266 cells, and the cytotoxic effects of CAR-BCMA-T cells were evaluated via CCK-8 and FACS. Finally, the CAR-BCMA-T cells originated from MM patients were constructed, and its cytotoxicity against A549-BCMA were examined; in addition, the IFN-γ release level in CAR-BCMA-T cells was evaluated by ELISA and FACS. Results: After 11 days’incubation, the CAR-BCMA-T cells originated from healthy donors amplified 300 times with a positive rate of 43%. The BCMApositive target cell lines were constructed successfully. Under an effector : target ratio of 5:1, the killing rates of CARBCMA-T cells against A549-BCMA, K562-BCMA and U266 were about 80%, 60%, and 80%, respectively, which were significantly higher than those against BCMA negative cells; and the cytotoxicity was related to the BCMA expression level in target cells. What’ s more, at the effector : target ratio of 20:1, the CAR-BCMA-T cells originated from MM patients were demonstrated to exhibit a killing rate of more than 95% againstA549-BCMApositive cells, and produced large amount of IFN-γ. Conclusion: CAR-BCMA-T cells originated from both healthy and MM donors were successfully constructed, and they can effectively and specifically kill BCMA positive tumor cells.

Influência de TLR3 em modelo de Paracoccidioidomicose experimental / TLR3 influence in experimental PCM model

Os receptores do tipo Toll compreendem a família de receptores de reconhecimento de padrões melhor caracterizados, que podem ativar diferentes respostas imunes, dependendo de quais receptores e conjuntos de adaptadores são utilizados. Os TLRs, como TLR2, TLR4 e TLR9, e sua sinalização foram implicados no reconhecimento de P. brasiliensis e na regulação da resposta imune, no entanto, o papel do TLR3 ainda não está claro. Assim, a compreensão da função endossomal do TLR3 na PCM experimental é crucial. Utilizamos modelos in vitro e in vivo de infecção por P. brasiliensis, camundongos C57Bl/6 e TLR3-/-, para avaliar a contribuição da TLR3 no desenvolvimento da infecção. Mostramos que ausência de TLR3 leva o aumento de óxido nítrico e a capacidade fagocítica por macrófagos nas primeiras 4 horas de interação com leveduras P. brasiliensis. Mostramos ainda que os camundongos TLR3-/- desempenham papel protetor após 30 dias de infecção intratraqueal com P. brasiliensis, mostrando diminuição do aumento de CFU, perfil de resposta Th1 e Th17, bem como aumento de células citotóxicas T CD8+ produtoras de IFN-γ e IL-17. As células citotóxicas T CD8+ mostraram ser essenciais para o controle da infecção nos camundongos TLR3-/-, uma vez que a depleção dessas células levou a progressão da doença. Em estágios iniciais, 3 e 5 dias de infecção, observamos aumento do recrutamento de neutrófilos para o pulmão. Estudos recentes indicam que o TLR3 é um receptor importante para a resposta imune na micose e sua ausência favorece a infecção por fungos. Em contraste, nossos resultados mostram que, no caso do PCM, o TLR3 é prejudicial ao hospedeiro, sugerindo que a ativação do TLR3 pode ser um possível mecanismo de escape de P. brasiliensis

Toll-like receptors comprise the best-characterized pattern-recognition receptor family that can activate different immune responses, depending on which receptor and adaptor set are utilized. TLRs, such as TLR2, TLR4 and TLR9, and their signaling have been implicated in the recognition of P. brasiliensis and regulation of the immune response, however, the role of TLR3 remains unclear. Thus, understanding the endosomal function of TLR3 in experimental PCM is crucial. We used in vitro and in vivo models of infection by P. brasiliensis, C57Bl/6 and TLR3-/- mice, to assess the contribution of TLR3 on development of infection. We show that absence of TLR3 leads to increased nitric oxide and phagocytic capacity by macrophages in the first 4 hours of interaction with yeasts P. brasiliensis. We also showed that TLR3-/- mice play a protective role after 30 days of intratracheal infection with P. brasiliensis, showing a decrease in the CFU increase, Th1 and Th17 response profile, as well as an increase in cytotoxic CD8+ cells producing IFN-γ and IL-17. The cytotoxic T CD8+ cells were shown to be essential for the control of infection in TLR3-/- mice, since the depletion of these cells led to the progression of the disease. In the initial stages, 3 and 5 days of infection, we observed increased recruitment of neutrophils to the lung. Recent studies indicate that TLR3 is an important receptor for the immune response in mycosis and its absence favors fungal infection. In contrast, our results show that in the case of PCM, TLR3 is detrimental to the host, suggesting that TLR3 activation may be a possible escape mechanism of P. brasiliensis

In Vitro Cytotoxicity Test and Surface Characterization of CoCrW Alloy in Artificial Saliva Solution for Dental Applications

Abstract In order to evaluate its application as a dental prosthesis material, a CoCrW alloy was subjected to in vitro cytotoxicity test, surface characterization and electrochemical studies performed in artificial saliva and 0.15 mol.L-1 NaCl medium. The used techniques were: anodic polarization curves, chronoamperometric measurements, electrochemical impedance spectroscopy (EIS), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS) analysis and X-ray photoelectron spectroscopy (XPS). Cytotoxicity test was also performed. The electrochemical behavior of CoCrW alloy was compared in both studied media, from corrosion potential (Ecorr) to a 600 mV anodic overvoltage. From the electrochemical measurements it was observed that the CoCrW alloy in both media presents only generalized corrosion. SEM and EDS analysis showed that the alloy presents carbide niobium and silicon and manganese oxides as nonmetallic inclusions. XPS results indicated that cobalt does not significantly contribute to the passivating film formation. Cytotoxicity test showed no cytotoxic character of CoCrW alloy. These results suggest that the CoCrW alloy can be used as biomaterial to be applied as prosthesis in dental implants.

Resumo Estudos eletroquímicos, caracterização de superfície e teste de citotoxicidade in vitro foram realizados da liga CoCrW em meios de saliva artificial e NaCl 0,15 mol.L-1, com o objetivo de avaliar a sua aplicação como material de prótese dentária. Foram usadas como técnicas, curvas de polarização anódica, medidas cronoamperométricas, espectroscopia de impedância eletroquímica (EIE), microscopia eletrônica de varredura (MEV), espectroscopia por energia dispersiva de raios X (EDS) e espectroscopia fotoeletrônica de raios X (XPS). O teste de citotoxicidade também foi realizado. O comportamento eletroquímico da liga CoCrW foi comparado nos dois meios estudados desde o potencial de corrosão (Ecorr) até uma sobretensão anódica de 600 mV. Foi observado, a partir de medidas eletroquímicas, que a liga CoCrW se encontra passivada em uma ampla faixa de potencial e que a sobretensões mais elevadas apresenta apenas corrosão generalizada nos dois meios. Análises por MEV e EDS mostraram que a liga apresenta inclusões não metálicas de carbeto de nióbio, de óxidos de silício e de manganês. Os resultados de XPS indicaram que o cobalto não contribui significativamente para a formação do filme passivo. O teste de citotoxicidade mostrou que a liga CoCrW não se apresenta citotóxica. Estes resultados sugerem que a liga estudada pode ser usada como biomaterial a ser aplicado como prótese sobre implantes dentários.

Bioactive lignans from Silybum marianum / 中国中药杂志

The seeds of Silybum marianum were extracted by hot water, and the extract was isolated by D101 macroporous resin, MCI resin, MPLC, HPLC, et al. As a result, 7 compounds including tricin 4'-O-[threo-β-guaiacyl-(7″-O-methyl)-glyceryl] ether(1), tricin 4'-O-[erythro-β-guaiacyl-(7″-O-methyl)-glyceryl] ether(2), 5'-methoxyhydnocarpin-D(3),palstatin(4),(8R,7'S,8'R)-5,5'-dimethoxy-7-oxolariciresinol 9'-O-D-xylopyranoside(5), 9-O-D-glucopyranoside(6), and(-)-haplomyrtoside(7) were isolated and identified for the first time. Compounds 1, 3, 4, and 5 exhibited activity against influenza A(H5N1)with IC₅₀ value of 0.65, 0.21, 0.32, and 0.56 μmol•L⁻¹, respectively. Compounds 1, 2, 6, and 7 exhibited cytotoxity against HepG-2 with IC₅₀ value of 0.35, 0.25, 0.53, 0.66 μmol•L⁻¹, respectively.

Cytotoxic triterpene glycosides from sea cucumber Holothuria leucospilota / 中国药学杂志

OBJECTIVE: To study the bioactive triterpene glycosides in sea cucumber Holothuria leucospilota. METHODS: Triterpene glycosides in the sea cucumber were isolated and purified by column chromatography on DA-101, silica gel, reversed-phase silica and reversed HPLC. Their chemical structures were identified on the basis of chemical properties and spectral data. The cytotoxicities of the compounds obstained were tested with human cervical carcinoma HeLa cells, gastric carcinoma BEL-7402 cells, and breast carcinoma MCF-7 cells. RESULTS: Three triterpene glycosides were obstained from the sea cucumber studied and their structures were identified as leucospilotaside D(1), holothurin B(2), and holothurin B2 (3). Their inhibition ratios for HeLa, BEL-7402, and MCF-7 cells were high. CONCLUSION: Compound 1 is obstained from Holothuria leucospilota for the first time. All glycosides show marked in vitro antitumor activity. Copyright 2012 by the Chinese Pharmaceutical Association.

Determinação da segurança biológica do xampu de cetoconazol: teste de irritação ocular e avaliação do potencial de citotoxicidade in vitro / Determination of the biological reactivity of ketoconazole in shampoo: draize eye test and cytotoxity test

Cetoconazol é um agente antifúngico, que pode ser incorporado em diferentes formas farmacêuticas, como, por exemplo, xampus e cremes. O objetivo do trabalho foi avaliar a segurança biológica in vivo (teste de irritação ocular) e in vitro (teste de citotoxicidade) do xampu de cetoconazol degradado sob ação da luz. Para tanto, a formulação foi exposta à radiação UV-C (254 nm) e foram empregados dois métodos para a determinação quantitativa do cetoconazol: CLAE e ensaio microbiológico. Os resultados demonstraram alteração do cetoconazol em presença da luz - presença de picos secundários no cromatograma e diminuição da atividade antifúngica - entretanto, não demonstraram alteração na segurança biológica entre xampu de cetoconazol e xampu de cetoconazol contendo produtos de degradação.

Ketoconazole is an antifungal agent and can be incorporated into several dosage forms, as an example it could be mentioned shampoos and creams. The aim of this work was to assess the biological reactivity in vivo (Draize eye test) and in vitro (cytotoxity test) of ketoconazole in shampoo degradeted under action of light. The formulation was exposed to UV-C (254 nm) radiation and two methods were used for the quantitative determination of ketoconazole: HPLC and microbiological assay. The results showed alteration in ketoconazole in presence of light - secondary peaks in chromatogram and decrease in antifungal activity - however, no alteration on the biological reactivity between ketoconazole shampoo and ketoconazole shampoo containing degradation products was observed.

Resistance to CTL-mediated cytotoxicity of target cells transfected with Wee1 / 中国免疫学杂志

Objective:To study the effect of resistance to CTL mediated cytotoxity of target cells transfected with Wee1 and treated with anti perforin antibody Methods:Cytotoxic T lymphocyte was obtained by mixing lymphocyte NIT culture in vitro,and proliferation index (PI) of lymphocytes was detected(MTT method) The recombinant vector pCMV Wee1Hu/GFP was transfected into mammalian cells NIT With the function of anti perforin antibody, NIT and NIT transfected with Wee1 used as target cells, CTL mediated cytotoxity was detected by LDH method Results:CTL proliferation was induced by mixing cell culture With anti perforin antibody, the cytolysis quantity of NIT transfected with Wee1 was least Conclusion:Target cells transfected with Wee1 could resist CTL mediated cytotoxity, and anti perforin antibody obviously increased the effect

The Induction of Cytokine-induced Killer and its Cytotoxic Effect on Hepatic Carcinoma Cells / 中国医师杂志

Objective To prepare cytokine-induced killer(CIK) in vitro, and study the biological activities of CIK. Methods Lymphocytes were isolated from peripheral blood and umbilical blood, and induced by cytokine.The cell surface markers CD_3 and CD_~56 of CIK were analyzed by FACS. The proliferation of CIK was tested using 3H -TdR release method, and the cytotoxic effect of CIK on hepatic carcinoma cells was measured by MTT. Results CIK cells quickly proliferated from the second week, and expanded more than 60-fold at the 31th day after induction. The CD_3+ and CD_~56 + cells expanded more than 800 folds. The cytotoxic effect of CIK on hepatic carcinoma cells was obviously stronger than that of LAK cells, and CIK had not obvious cytotoxic effect on fetal liver cells. Conclusion CIK was a highly efficient cytotoxic cells against tumors, and had clinical application potentials.

Cardiotoxicity of ketamine in isolated myocardial cells of neonatal rat / 中华麻醉学杂志

Objective: To investigate ketamine cardiotoxicity profile. Method:Four day-old spontaneously contracting neonatal primary myocardial cell cultures obtained from 2-to 3-day-old Wistar rats were divided into 4 groups, group A as control and group B,C and D treated with ketamine(1?10~(-5), 1?10~(-4)and 1?10~(-3)mol/L)for 2 to 24 h. The contractility, morphology,cytoplasmic enzyme (LDH, AST and CK) release content of myocardial cell and the concentration of electrolytes (k~+, Na~+, Ca~(2+) and Cl~-) in the medium were measured 2,4,8 and 24 h following ketamine administration. Result:In group B the beating rates of neonatal myocardial cell cultures increased (P

Studies on T cell,T Cell Subpopulation and Natural Cell Activities of the Peripheral Blood Lymophocytes from Patients with Tumor / 中国免疫学杂志

In this paper,we reported that T cell,T cell subpopulation and natural killer(NK)cell activities of peripheral blood lymophocytes from the Fatients with tumor weredetected with OKT monoclonal antibody and ~(51)Cr release cytotoxity test.The resultsshowed that percentage of OKT3~+,OKT4~+ cell in the peripheral blood of patients withtumor are low,OKT8~+ is high and the ratio of OKT4/OKT8 is low.The results haveproved that the ratio of OKT4/OKT8 and NK cell activities were significantly low inthe peripheral blood of patients with tumor.However,the activities have restored tonormal or near normal level in the patients after surgical operation for more that twoyears with no recurrence. Our data suggest that detecting of the ratio of OKT4/OKT8 and activities of NKcell might be valuable to the prognosis of patients with tumor.