Concomitant using of topical carrageenan-kappa and oral vitamin D against 7, 12-dimethylbenz[a]anthracene induced-oral cancer in rats: A synergism or an antagonism effects / Uso concomitante de carragenina tópica e vitamina D oral contra câncer de boca em ratos induzido por 7, 12-dimetilbenz(a) antraceno: efeitos sinérgicos ou antagonistas

Objective: κ-carrageenan is a food stabilizer agent which has an antiproliferative effect, while vitamin D is a prohormone acts on the nuclear receptor and has a cytotoxic against cancer. This study aimed to show the synergistic effect of using topical κ-carrageenan and oral administration of the vitamin D on the 7, 12-dimethylbenz[a] anthracene (DMBA)-induced oral cancer. Material and Methods: fifty four male albino rats were randomly divided into seven groups: Acetonetreated served as control (Group I), vitamin D (5000UI)-treated (Group II), κ-carrageenan (1%)- treated (Group III), DMBA (0.5%)-treated (Group IV), Acetone, κ-carrageenan and DMBA were administered topically on both cheeks and palate, five times weekly for 12 weeks, while the vitamin D was administered orally twice weekly for 12 weeks. Groups V, VI, and VII were animals treated with vitamin D, κ-carrageenan, and both vitamin D and κ-carrageenan for 8 weeks after induction of oral cancer. At the end of the study, blood samples were obtained by cardiac puncture for determination of TNF-α and EGFR. Results: In the groups III and IV, serum EGFR showed significant low levels compared with Group I. In the Group VII, serum EGFR showed a significantly (p=0.014) low level compared with Group IV (614.3±69.7 pg/ml versus 882.4±45.6 pg/ml, respectively). Higher percentages of high levels of TNF-α were observed in the Groups VI and VII, while a lower percentage of EGFR was observed in the Group VI. Conclusion: both κ-carrageenan and vitamin D have antiproliferative effect against DMBAinducing oral cancer by increasing the levels of TNF-α and suppressing the signaling pathway of EGFR. Concomitant using κ-carrageenan and vitamin D reduces the antiproliferative effect of each other.(AU)

Objetivo: κ-carragenina é um agente estabilizador de alimentos que tem efeito um antiproliferativo, enquanto a vitamina D é um pró-hormônio que atua sobre o receptor nuclear e possui efeito citotóxico contra o câncer. Este estudo teve como objetivo mostrar o efeito sinérgico do uso de κ-carragenina tópica e administração oral da vitamina D no câncer de boca induzido por 7, 12-dimetilbenz[a]antraceno (DMBA). Material e Métodos: cinquenta e quatro ratos albinos machos foram divididos aleatoriamente em sete grupos: tratado com acetona como controle (Grupo I), tratado com vitamina D (5000UI) (grupo II), tratado com κ-carragenina (1%) (grupo III), DMBA (0,5%) tratado (Grupo IV), acetona, κ-carragenina e DMBA foram administrados topicamente nas bochechas e no palato, cinco vezes por semana durante 12 semanas, enquanto a vitamina D foi administrada por via oral duas vezes por semana durante 12 semanas. Os grupos V, VI e VII foram animais tratados com vitamina D, κ-carragenina e No final do estudo, foram obtidas amostras de sangue por punção cardíaca para determinação do TNF-α e EGFR. Resultados: Nos grupos III e IV, o EGFR sérico mostrou níveis baixos significativos em comparação com o Grupo I. No grupo VII, o EGFR sérico mostrou um nível significativamente baixo (p = 0,014) em comparação com o Grupo IV (614,3 ± 69,7 pg / ml versus 882,4 ± 45,6 pg / ml, respectivamente). Maiores porcentagens de TNF-α foram observadas nos Grupos VI e VII, enquanto uma menor porcentagem de EGFR foi observada no Grupo VI. Conclusão: Tanto a κ-carragenina quanto a vitamina D têm efeito antiproliferativo contra o câncer de boca induzido por DMBA aumentando os níveis de TNF-α e suprimindo a via de sinalização do EGFR. O uso concomitante de κ-carragenina e a vitamina D reduz o efeito antiproliferativo um do outro (AU)

Comparison of modeling effects of two different 7, 12-dimethylbenza anthracene induced breast cancer models in tree shrew / 中华肿瘤杂志

Objective@#To explore the feasibility of 7, 12-dimethylbenz[a] anthracene (DMBA) induced tree shrew breast cancer model, and compare the effects of two administration modes by gavage and mammary gland injection.@*Methods@#A total of 40 tree shrews were randomly divided into two groups (20 animals per group): DMBA gavage group and mammary gland injection group. DMBA was dissolved in edible vegetable oil. For gavage group, tree shrews were administered with DMBA solutions (15 mg/kg) by gavage once a day. For mammary gland injection group, DMBA solution (10 mg/kg) was injected into the mammary fat pad of tree shrews, and the injection was performed for a total of 3 times. From the first administration of DMBA, medroxyprogesterone acetate (MPA, 100 mg/kg) was intramuscularly injected into the muscles of the lateral thighs of tree shrews at the same time, for a total of 5 times. The tumorigenesis and survival of tree shrews were monitored. The tumor histological morphology was observed by HE staining. The expression of estrogen receptor (ER), progesterone receptor (PR), cytokeratin5/6 (CK5/6) and human epidermal factor receptor-2 (HER-2) was detected by immunohistochemical staining.@*Results@#In the gavage group, there were 10 deaths, and 4 tree shrews developed mammary tumors with 20.0% (4/20) tumor formation rate. The success rate of mammary cancer modeling was 10.0% (2/20), and the tumor formation time was 197.3±15.1 days. In the mammary gland injection group, there were 8 tree shrews died, and 9 tree shrews formed tumors with 45.0% (9/20) tumor formation rate. The success rate of mammary cancer modeling was 40.0% (8/20), and the tumor formation time was 71.8±19.0 days. There was no significant difference in mortality and tumor formation rate (P>0.05) between the two groups (all P>0.05). However, in the mammary gland injection group, the success rate of mammary cancer modeling was significantly higher than that in the gavage group (P<0.05), whereas the tumor formation time was markedly shorter than that in the gavage group (P<0.01). The pathological types in the gavage group included ductal hyperplasia, intraductal papilloma and ductal carcinoma in situ, while those in the breast injection group included intraductal papilloma and ductal carcinoma in situ. In both groups, immunohistochemical staining showed the negative expression of HER-2 but positive expression of ER, PR and CK5/6 with varying degrees.@*Conclusion@#Both the DMBA gavage and mammary gland injection can successfully establish the tree shrew breast cancer model, and the modeling effect of mammary gland injection is better than gavage.

Research progress on chemical composition and pharmacological effects of Rhei Radix et Rhizoma and predictive analysis on quality markers / 中草药

Rhei Radix et Rhizoma is a kind of traditional Chinese medicinal material in China, which has unique efficacy and rich resources. It is mainly distributed in the alpine mountains of temperate and subtropical Asia. Its genuine producing areas are Gansu Province, Qinghai Province, Sichuan Province, and Tibet Autonomous Region. In recent years, with the continuous in-depth study of Radix et Rhizoma by scholars at home and abroad, many kinds of chemical constituents, extensive pharmacological effects and application and development of Radix et Rhizoma have attracted much attention, and have broad application prospects and considerable market value. Based on the analysis of its chemical composition and traditional efficacy combined with modern pharmacological action, and according to the theory of quality marker, the quality markers of Radix et Rhizoma were predicted and analyzed from the aspects of the chemical composition and traditional medicinal properties, traditional efficacy, new clinical uses, plasma composition, measurable components, effective components in different compatibility, and the effect of storage time and so on, which provides a scientific basis for the quality evaluation of Radix et Rhizoma and establishment of its quality control system.

Direct large-volume injection analysis of polycyclic aromatic hydrocarbons in water / Análisis de inyección directa en gran volumen de hidrocarburos aromáticos policíclicos en agua / Análise de injeção direta de grande volume de hidrocarbonos aromáticos policíclicos em água

Abstract Due to the health risks for both humans and living beings caused by polycyclic aromatic hydrocarbons (PAHs), the monitoring of these compounds in environmental matrices is mandatory. This work proposes an analytical method for analyzing anthracene (AN) and benzo[a]pyrene (BaP), two of the most representative PAHs, at ultra-trace concentrations in water, employing direct injection of large volumes of samples coupled with reversed-phase high-performance liquid chromatography. For this purpose, principal component analysis was used to examine the behavior of AN and BaP within the chromatographic system. Results showed that AN and BaP chromatographic behavior can be described by three models representing their identification, the quantification of AN and that of BaP, respectively. The factors affecting the obtained models, such as the injection volume, column temperature, flow rate, strength of the mobile phase, and the excitation and emission wavelengths, were examined and optimized by means of design of experiments. Finally, the analytical method was validated, obtaining promising limits of detection and quantification. The developed analytical method was demonstrated to be useful for a sensitive analysis of the target analytes in relatively clean natural water matrices.

Resumen Los hidrocarburos aromáticos policíclicos (HAPs) causan problemas en la salud de los seres humanos y seres vivos, por lo que se requiere un monitoreo de estos compuestos en matrices ambientales. Este trabajo propone un método analítico para analizar el antraceno (AN) y el benzopireno (BAP), los hidrocarburos más representativos en concentraciones de ultra trazas en el agua, empleando inyección directa en grandes volúmenes en muestras acopladas a la fase inversa con cromatografía líquida de alto rendimiento. Por tal razón, se utilizó el análisis de componentes principales para examinar el comportamiento de AN y BAP en el sistema cromatográfico. Los resultados mostraron que el comportamiento cromatográfico de AN y BAP podría describirse por medio de tres modelos que representan su identificación, la cuantificación de AN y de BAP, respectivamente. Se examinaron los factores que afectan a los modelos obtenidos, como el volumen de inyección, la temperatura de la columna, la tasa de flujo, la fuerza de la fase móvil, y las longitudes de las ondas de excitación y emisión, y se optimizaron mediante el diseño de experimentos. Finalmente, se validó el método analítico, obteniendo límites de detección y cuantificación. Se demostró que el método analítico desarrollado fue útil para el análisis sensible de los analitos en matrices de agua natural relativamente limpia.

Resumo Devido aos riscos para a saúde tanto para humanos como para os seres vivos em geral causados pelos hidrocarbonos aromáticos policíclicos (HAPs), o monitoramento de estes compostos em matrizes ambientais é prioritário. Este trabalho propõem um método analítico para analisar antraceno (AN) e benzo[a]pireno (BaP), dois dos hidrocarbonos mais representativos, em concentrações de ultra traços em água, empregando injeções diretas de grandes volumes de amostra acoplada a cromatografia líquida de alta eficiência em fase reversa. Usando Análises por Componentes Principais e desenho experimental, foram avaliados os efeitos de diversos fatores que afetam o sistema cromatográfico, tais como o volume de injeção, a temperatura da coluna, fluxo, forca da fase móvel e comprimentos de onda de excitação e emissão. Os resultados demonstraram que o comportamento cromatográfico de AN e BaP pode ser descrito por meio de três que representam sua identificação, quantificação de AN e de BaP, respectivamente. Os resultados mostraram que o comportamento cromatográfico de NA e BAP poderia ser descrito por meio de três modelos que representam sua identificação, a quantificação de NA e de BAP, respectivamente. Examinaram-se os fatores que afetam aos modelos obtidos, como o volume de injeção, a temperatura da coluna, a taxa de fluxo, a forca da fase móvel, e as longitudes das ondas de excitação e emissão, e se otimizaram mediante o desenho experimental. Finalmente, se validou o m todo analítico, obtendo os limites de detecção e quantificação. O método analítico desenvolvido demonstrou ser útil para uma análise sensível para os compostos de interesse em matrizes de água natural relativamente limpas.

Presence of Eisenia fetida enhanced phytoremediation of anthracene by Lolium perenne / Presença de Eisenia fetida aumentou a fitorremediação do antraceno por Lolium perene

The efficiency of co-application of Eisenia fetida and ryegrass was evaluated in a process called earthworm-assisted phytoremediation. Anthracene was used as a model compound for polycyclic aromatic hydrocarbons (PAHs). The experiments were conducted on a loamy soil in greenhouse conditions. At the end of the experiment, the soil samples were analyzed for residual anthracene by HPLC. Results showed that, phytoremediation using ryegrass could remove 81% of anthracene; however, the rate of removal was 92% when E. fetida was applied simultaneously. E. fetida alone could also remove the initial concentration of anthracene by 40%. Although ryegrass itself could remove anthracene significantly, the employment of earthworm, together with plant was more efficient than each of them individually. The application of E. fetida could also enhance the growth parameters of ryegrass significantly. In comparison to the control, the presence of E. fetida increased plant dry weight (7.8%), root length (47%), shoots length (32%), and root volume (12%). The number of live earthworms was also increased in the planted pots, indicating the helpfulness of the plant for survival of the earthworm in the PAH-contaminated soil. Although plant and earthworm use completely different mechanisms for anthracene degradation, they improve efficiency and survival of the three-component-system.

A eficiência da co-aplicação de Eisenia fetida e azevém foi avaliada em um processo denominado fitorremediação assistida por minhocas. O antraceno foi usado como um composto modelo para hidrocarbonetos aromáticos policíclicos (PAHs). Os experimentos foram conduzidos em um solo argiloso em condições de estufa. No final da experiência, as amostras de solo foram analisadas quanto ao antraceno residual por HPLC. Os resultados mostraram que, a fitorremediação com azevém pode remover 81% do antraceno; no entanto, a taxa de remoção foi de 92% quando E. fetida foi aplicada simultaneamente. E. fetida sozinha também foi capaz de remover a concentração inicial de antraceno em 40%. Embora o próprio azevém pudesse remover significativamente o antraceno, o emprego da minhoca, juntamente com a planta, foi mais eficiente do que cada um deles individualmente. A aplicação de E. fetida também pode melhorar significativamente os parâmetros de crescimento do azevém. Em comparação com o controle, a presença de E. fetida aumentou o peso seco da planta (7,8%), o comprimento da raiz (47%), o comprimento da parte aérea (32%) e o volume radicular (12%). O número de minhocas vivas também aumentou nos vasos plantados, indicando a utilidade da planta para a sobrevivência da minhoca no solo contaminado com PAH. Embora plantas e minhocas usem mecanismos completamente diferentes para a degradação do antraceno, eles melhoram a eficiência e a sobrevivência do sistema de três componentes.

Simultaneous detection of 3 kinds of polycyclic aromatic hydrocarbons in the air of workplace by high performance liquid chromatography / 中国职业医学

OBJECTIVE: To establish a method for simultaneous detection of 3 kinds of polycyclic aromatic hydrocarbons(PAHs) including phenanthrene,anthracene and 3,4-benzo( a) pyrene in workplace air by high performance liquid chromatography( HPLC). METHODS: The phenanthrene,anthracene and 3,4-benzo( a) pyrene were collected in the air of the workplace using glass fiber filter paper. The membrane sample was added with 5. 00 m L of acetone,and the samples were extracted by ultrasonic wave for 70 mintues and eluted with acetonitrile-water gradient. Liquid chromatography-UV detector with tandem fluorescence detector was used for determination. RESULTS: The good linearity rang of phenanthrene,anthracene and 3,4-benzo( a) pyrene was 0. 050-1. 000 mg/L with the correlation coefficients ≥0. 999 5; the detection limits were 0. 010,0. 007 and 0. 008 mg/L,and the minimum detectable concentrations were 0. 130,0. 090 and 0. 110μg/m3 respectively( air collection of 375 L). The desorption efficiencies were 86. 10%-99. 20% of phenanthrene,88. 60%-96. 40% of anthracene,and 84. 80%-99. 60% of 3,4-benzo( a) pyrene,respectively. The within-run relative standard deviation( RSD) were 1. 29%-3. 25%,1. 90%-3. 61% and 1. 30%-4. 82% respectively,the between-run RSD were 2. 41%-4. 07%,2. 02%-5. 12% and 2. 08%-4. 77% respectively. The standard recovery rates were 95. 00%-106. 14% of phenanthrene,93. 14%-106. 50% of anthracene, and 92. 86%-105. 50% of 3,4-benzo( a) pyrene,respectively. The samples could be stored at 4 ℃ for 7 days. CONCLUSION: This method is simple,accurate and reliable for the simultaneous detection of phenanthrene,anthracene and 3,4-benzo( a) pyrene in the air of workplace.

Dampness-Heat Accelerates DMBA-Induced Mammary Tumors in Rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the impact of dampness-heat (DH) on the development of mammary tumors in 7,12-dimethylbenz(a)anthracene (DMBA)-induced rats.</p><p><b>METHODS</b>Forty rats were randomly divided into 3 groups in a randomized block design, including the control group (n=13), DMBA group (n=14), and DMBA plus DH group (n=13). Rats in the DMBA group and DMBA plus DH group were intragastrically administrated with DMBA (100 mg/kg) for twice, once per week, while rats in the control group were treated with equivalent volumes of sesame oil. After DMBA administration, rats in the DMBA plus DH group were exposed to a simulated climate chamber with ambient temperature (33.0±0.5°C) and humidity (90%±5%) for 8 weeks, 8 h per day. The body weight, time of tumor formation, and number of tumors were measured weekly to calculate tumor incidence, average latency period, average number of tumors, and average tumor weight. At the end of the experiment, the levels of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinases 1 (TIMP-1) in serum, and the contents of tumor necrosis factor-α (TNF-α) and interleukin (IL)-1β in serum and tumor tissue were measured, respectively. Some tumor tissues were processed for hematoxylin-eosin staining to determine the histopathological changes.</p><p><b>RESULTS</b>Compared with DMBA, DMBA plus DH significantly increased the average number of tumors, average tumor weight, levels of serum MMP-9, TIMP-1, TNF-α and IL-1β, and contents of tumor tissue TNF-α and IL-1β (P<0.05 or P<0.01).</p><p><b>CONCLUSION</b>DH could accelerate the development of mammary tumors through increasing the expressions of MMP-9, TIMP-1, TNF-α and IL-1β in DMBA-induced rats.</p>

Genotoxicidad de los hidrocarburos aromáticos policíclicos extraídos mediante el sistema diclorometano-etanol-tolueno en muestras del aire de Cúcuta, Norte de Santander, Colombia / Genotoxicity of polycyclic aromatic hydrocarbons extracted by dichloromethane-ethanol-toluene system air samples from Cúcuta, Norte de Santander, Colombia

Los contaminantes del aire han sido y siguen siendo, los principales factores que contribuyen a las enfermedades crónicas como el asma y enfermedades cardiovasculares. La contaminación del aire por material particulado (PM) es un problema mundial y en los últimos años, el PM se ha convertido en un tema importante de investigación ya que tiene un impacto negativo significativo en la salud humana; el PM es generado por las actividades industriales y tubos de escape de vehículos de motor. Sin embargo, diversos componentes nocivos del PM, como los hidrocarburos aromáticos policiclicos (HAP) en general, son sos­pechosos de ser carcinogénicos. Este trabajo tiene como objetivo identificar los HAP presentes en el PM2.5 del aire de Cúcuta, extraídos por primera vez, mediante el sistema diclorometano-etanol-tolueno e investigar la importancia del fraccionamiento de la materia organica del PM2.5 para detectar los HAP presentes en las fracciones del PM2.5. La identificación de los HAP considerados como contaminantes prioritarios y reconocidos por su afectación a la salud de la población se realizó, mediante cromatografía de gases con detector FID. Los efectos genotoxicos de la materia orgánica del PM2.5 extraída con una mezcla de DCM-etanol-tolueno fueron evaluados mediante el ensayo Cometa.

Air pollutants have been and still are the main factors that contribute to chronic diseases such as asthma and cardio­vascular disease. Air pollution by particulate matter (PM) is a global problem and in recent years, the PM has become an important research topic since it has a significant negative impact on human health; the PM is generated by industrial activities and exhaust pipes of motor vehicles. However, various harmful components of PM such as polycyclic aromatic hydrocarbons (PAHs) in gen­eral, are suspected of being carcinogenic. This work aims to identify the PAHs present in the PM 2.5 air Cúcuta, first extracted by the dichloromethane-ethanol-toluene system and investigate the importance of organic matter fractionation of PM 2.5 to detect PAHs present in the fractions of PM 2.5. The identification of PAHs considered as priority pollutants and recognized for their effects on health of the population was performed by gas chromatography with FID detector. The genotoxic effects of PM2.5 organic mat­ter, extracted with a mixture of DCM-ethanol-toluene, was evaluated by the Comet assay.

9-[(E)-2-feniletenil]antraceno Y 9-[(E)-2-(naftalen-2-il)etenil] antraceno COMO TRAMPAS PARA OXÍGENO SINGULETE: OXIDACIÓN FOTOSENSIBILIZADA Y EFECTO FOTODINÁMICO SOBRE PARÁSITOS Leishmania tarentolae / 9-[(E)-2-phenylethenyl]anthracene AND 9-[(E)-2-(naphthalen-2-yl)ethenyl] anthracene AS TRAPS FOR SINGLET OXYGEN: PHOTOSENSITIZED OXIDATION AND PHOTODYNAMIC EFFECT ON Leishmania tarentolae PARASITES

Introducción: El oxígeno singulete es una especie reactiva que se obtiene mediante transferencia energética usando un fotosensibilizador. Su cuantificación directa requiere de instrumentación costosa, por lo cual es necesario recurrir a métodos indirectos que tengan suficiente selectividad y bajo costo. Estos procedimientos se basan en la interceptación química del oxígeno singulete produciendo una especie que se pueda detectar por métodos analíticos convencionales. En este artículo se describe la utilización del 9-[(E)-2-feniletenil] antraceno 1 (PEA) y del 9-[(E)-2-(naftalen-2-il) etenil]antraceno 2 (NEA), como alternativas viables y económicas para la cuantificación indirecta del oxígeno singulete, en medios acuosos. Su ventaja radica en la fácil detección de la desactivación de su fluorescencia una vez son oxidados por el oxígeno singulete. Materiales y Métodos: Los compuestos se sintetizaron y caracterizaron siguiendo procedimientos previamente reportados. Su capacidad para atrapar oxígeno singulete se determinó siguiendo su oxidación fotosensibilizada en solución de H2O/THF y en parásitos de Leishmania tarentolae, empleando azul de metileno o rosa bengala como fotosensibilizadores. Las muestras experimentales se iluminaron con una lámpara de emisión de luz visible, y se utilizaron métodos espectroscópicos (absorción UV-Vis, fluorescencia, RMN-1H) y espectrometría de masas para monitorear el atrapamiento y fotooxidación. Resultados y Discusión: Las pruebas espectroscópicas demostraron la capacidad que tienen los compuestos PEA 1 y NEA 2 para atrapar oxígeno singulete en solución acuosa y dentro de parásitos de L. tarentolae. Estudios de viabilidad parasitaria demuestran que PEA 1 es citotóxico en la oscuridad y cuando los cultivos son expuestos a la luz, mientras que NEA 2 no es citotóxico en la oscuridad, pero sí lo es cuando el cultivo es expuesto a la luz. En conclusión, los compuestos estudiados pueden servir como sondas para detectar y medir la producción de oxígeno singulete en medio acuoso y potencialmente en cultivos celulares, aunque es recomendable evaluar su actividad citotóxica en la oscuridad y bajo iluminación en estos casos.

Introduction: Singlet oxygen is a reactive species obtained via energy transfer using a photosensitizer. Its direct quantification requires expensive instrumentation, so it is necessary to use indirect methods having sufficient selectivity and low cost. These procedures are based on the chemical interception of singlet oxygen producing a species that can be detected using conventional analytical methods. This article describes the utilization of 9-[(E)-2-phenylethenyl]anthracene 1 (PEA) and 9-[(E)-2-(naphtalen-2-yl)ethenyl]anthracene 2 (NEA) as suitable and economic alternatives for the indirect quantification of singlet oxygen in aqueous media. Their advantage is the easy detection of their fluorescence once they are oxidized by singlet oxygen. Materials and Methods: Compounds were synthesized and characterized following procedures previously reported. Their capacity to trap singlet oxygen was determined by monitoring their photosensitized oxidation in either a H2O/THF solution or within Leishmania tarentolae parasites, utilizing methylene blue or rose bengal as photosensitizers. Experimental samples were illuminated with a lamp emitting visible light, while spectroscopical techniques (absorption, fluorescence, 1 H-NMR) and mass spectrometry were used to monitor trapping and photooxidation. Results and Discussion: Spectroscopical evidence demonstrates that both PEA 1 and NEA 2 are capable of trapping singlet oxygen in both aqueous media and within L. tarentolae parasites. Viability studies demonstrate that PEA 1 is cytotoxic in the dark and when parasite cultures were exposed to light, while NEA 2 does not show dark cytotoxicity, but is toxic when cultures were exposed to light. It can be concluded that both compounds under study may be utilized as probes to detect and quantify the production of singlet oxygen in aqueous media and potentially in cell cultures, although it is recommended to evaluate their cytotoxic activity both in the dark and upon light exposure in these cases.

Effects of focused ultrasound on grade i cervical intraepithelial neoplasia in mice / 肿瘤

Objective: To investigate the effect of focused ultrasound on murine cervical intraepithelial neoplasia (CIN) and its possible mechanism. Methods: The mice bearing grade I CIN were orthotopically induced by dimethl benz anthracene and randomly divided into treatment group (n = 20) and control group (n = 20). The mice in the treatment group were treated with focused ultrasound, while the mice in the control group were treated with the same method but without power output. After two weeks, the pathological changes and the expressions of p16, Ki67, FasL and Fas in murine cervical tissues were detected by HE staining and immunohistochemistry, respectively. Results: The model of grade I CIN in mice was successfully established. In the treatment group, the cervical tissues returned to normal in 15 mice after treatment, 4 mice remained grade I CIN, and 1 mouse died. The response rate of treatment group was 75% (15/20). In the control group, the cervical tissues remained grade I CIN in 11 mice after treatment, 6 mice developed to grade II CIN, 1 mouse developed to grade III CIN, and 2 mice died. The response rate of the control group was 0% (0/20). There was a significant difference between the treatment group and the control group (P < 0.01). The strong positive expression rates of p16, Ki67 and FasL in cervical tissues in the treatment group were lower than those in the control group, while the strong positive expression rate of Fas was opposite (all P < 0.01). Conclusion: The focused ultrasound can effectively treat the grade I CIN in mice. This effect may be related to the changes of p16, Ki67, Fas and FasL expressions.

The research on apoptosis of human laryngeal cell line Hep-2 induced by 1,4-bis[2-(benzylselanyl) ethoxy] anthracene / 中国癌症杂志

Background and purpose:Selenium is one of the essential trace elements for human activities, and plays an incomparable role in maintaining human health. It was reported that selenium compound 1,4-bis[2-(benzylse-lanyl)ethoxy] (BSEA) anthracene has antiseptic and antiphlogistic effects. However, the mechanisms underlying anti-cancer effects of BSEA are rarely reported. BSEA-induced apoptosis in human laryngeal carcinoma Hep-2 cells and its mechanisms were studied.Methods:Methyl thiazolyl tetrazolium (MTT) assay was used to determine inhibition ratio of Hep-2 cells 24 hours after Hep-2 cells were treated with different concentrations of BSEA. Fluorescence microscope was used to observe the morphology change of apoptosis in Hep-2 cells. The apoptosis was detected by Annexin Ⅴ-FITC. Mi-tochondrial membrane potential was assayed by JC-1. Microplate reader detected the activity of caspase-3 and caspase-8. The mRNA and protein levels of Bax and XIAP were measured by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot.Results:The results showed that BSEA caused a dose-dependent inhibition of the growth of human laryngeal carcinoma cell line Hep-2in vitro, andIC50 was 35.74μmol/L. The apoptotic bodies were distinctly observed at a concentration of 80μmol/L of BSEA by AO fluorescence staining. This study found that the eversion of phosphatidyl serine intensified, and mitochondrial membrane potential also began to decline. The activity of caspase-3 appeared the tendency of dependence on dosage, while the activity of caspase-8 did not change significantly. The mRNA and protein expression level of Bax increased, whereas the mRNA and protein expression level of XIAP de-creased.Conclusion:Therefore, BSEA could obviously inhibit human laryngeal carcinoma Hep-2 cells proliferation and induce apoptosis via the mitochondrial pathway.

In situ Determination of Anthracene Adsorbed onto Mangrove Root Surface Micro-zone Using Microscopic Fluorescence Spectrometric Analysis System / 分析化学

A microscopic fluorescence spectrometric analysis ( MFSA) system was set in the laboratory. A novel method for in situ determination of Anthracene ( Ant) adsorbed onto root surface micro-zone of two kinks of mangrove plant, named Kandelia obovata ( K. obovata) and Avicennia marina ( A. marina) by MFSA was established. Fluorescence spectra of Ant adsorbed on root surface micro-zone were obtained by synchronous scanning mode. The signal to noise (S/N) of Ant (5. 3 pg/μm2) adsorbed on K. obovata and A. marina root surface micro-zone increased up to 5. 5 and 6. 8 while wavelength offset (△λ) both were at 60 nm, respectively. The linear dynamic ranges of established method were 5. 3-63. 2 pg/μm2 for K. obovata and 10. 5-52. 6 pg/μm2 for A. marina, with the detection limits of 1. 1 pg/μm2 and 5. 5 pg/μm2 , respectively. The relative standard deviations were both less than 12 . 5% ( n=9 ) , and the recoveries were 98 . 1% -117. 0% and 81. 2%-110. 9%, respectively. The result showed that the MFSA system had ability to obtain quantitative information of fluorescence spectra and fluorescence image of Ant adsorbed onto plant roots surface micro-zone.

Establishment of a mouse model of cutaneous squamous cell carcinoma induced by 7,12-DMBA, croton oil and narrow-band ultraviolet B irradiation / 中国比较医学杂志

Objective To establish a mouse model of cutaneous squamous cell carcinoma induced by 7,12-dimethylbenz(a)anthracene (7,12-DMBA)/croton oil and narrow-band ultraviolet B (NB-UVB) irradiation.Methods A total of fifty 6-8-week old BALB/c mice (male:female 1:1) were randomly divided into three experimental groups.The group A was treated with chemical carcinogens alone, group B was treated with NB-UVB alone, and group C was treated with chemical carcinogens plus NB-UVB.The general status and skin appearance of mice were observed during the experiment.The survival rate and tumor formation rate of each group was calculated at weeks 5, 10, 15, and 20. Pathological examination was carried out to observe the histological changes of skin lesions.Results Papules measuring≥l mm in diameter began to develop in some mice of the group C at 5 weeks after the first treatment with chemical carcinogens.The tumor formation rates at 20 weeks after treatment were 86.67%, 7.14%, 94.12%in the groups A, B, C, respectively.Pathological examination revealed characteristic changes of squamous cell carcinoma in 13.34%, 0%, 70.59%of the mice in the group A, B, C, respectively.Conclusions Establishment of a mouse model of cutaneous squamous cell carcinoma induced by 7,12-DMBA/croton oil and NB-UVB is a better method than treated with chemical carcinogens alone or NB-UVB alone.This method can increase the tumor formation rate and incidence rate of SCC, and within a shorter period.

Evaluation of anti-tumor activity of Momordica cymbalaria fenzl.

Background: The aim of the current study is to evaluate the anti-tumor activity of saponins isolated from the roots of Momordica cymbalaria (MC) against dimethylbenz[a]anthracene (DMBA) induced rats. Methods: A steroidal saponin MC (SMC) was isolated from MC fenzl and purified by preparative high-performance liquid chromatography. Breast cancer was induced in 50-day-old female Sprague-Dawley rats by injecting DMBA (6 mg/kg intravenous) in three doses on day 50, 54, and 57. The rats were randomized into four groups; control, DMBA, SMC (100 mg/kg), and tamoxifen (6.6 mg/kg) to DMBA breast cancer rats. The tumor size, volume, hormonal, antioxidant, and whole mount parameters were estimated. Results: Mean tumor size and volume, luteinizing hormone, and progesterone with superoxide dismutases, catalase, and glutathione levels increased significantly (p<0.001); serum estradiol, follicle stimulating hormone with lipid peroxidation decreased significantly (p<0.001) in DMBA-induced breast cancer and vice versa in SMC and tamoxifen. Terminal end buds, terminal ducts, alveolar buds, and lobules decreased significantly (p<0.001) in DMBA-induced breast cancer whereas increased significantly in SMC and tamoxifen. Histological necrosis and hemorrhage along with focal desmoplastic reaction in DMBA-induced breast cancer; ductile elongation and hyperplasia of both ducts and alveoli were prominent, with increased secretory activity in SMC group. The results confirmed the chemopreventive effect of SMC and tamoxifen in DMBA-induced breast cancer. Conclusions: The SMC exhibited anti-tumor activity against mammary cancer, which may be due to its anti-estrogenic, antioxidant activity.

Synthesis and antibacterial activity of some 3-(5-aryl-4H-pyrazol-3-yl).

A series of 3-(5-aryl-4H-pyrazol-3-yl)anthracen-10(9H)-ones were synthesized from anthracen-10(9H)-one (1) and studied for their in vitro antibacterial activity. Anthracen- 10(9H)-one after Friedel crafts acetylation with acetyl chloride yielded 3-acetylanthracen- 10(9H)-one (2) which on further reaction with substituted aromatic aldehydes in the presence of catalytic amount of sodium hydroxide in water and ethanol furnished the corresponding 3-(3-arylacryloyl)anthracen-10(9H)-ones (3a-g) as intermediate compounds, which on further reaction with hydrazine hydrate in absolute ethanol formed the title compounds 3-(5-aryl-4H-pyrazol-3-yl)anthracen-10(9H)-ones (4a-g). These compounds were characterized by elemental analysis, IR, Mass and 1H-NMR spectral data. All the compounds were evaluated for their in vitro antibacterial activity against two gram positive strains (Bacillus subtilis and Staphylococcus aureus) and two gram negative strains (Escherichia coli and Pseudomonas aeruginosa) taking ciprofloxacin as a standard drug. Some of the compounds showed significant antibacterial activity.

Azadirachta indica mitigates DMBA-induced hepatotoxicity: A biochemical and radiometric study.

The hepatoprotective potential of aqueous Azadirachta indica leaf extract (AAILE) was assessed against DMBA-induced hepatotoxicity. DMBA  (7,12-dimethylbenz[a] anthracene) treatment (40 mg/kg body weight, ip) to male Balb/c mice resulted in the derailment of liver function as revealed by extremely slow clearance of 99mTc-mebrofenin from liver, elevated levels of alkaline phosphatase (ALP) and alanine transaminase (ALT), compared to control group. In addition, elevated micronuclei score and high apoptotic index indicated hepatogenotoxicity in DMBA-treated mice. DMBA treatment also upregulated cytochrome P450 (CYP), cytochrome b5 (Cyt b5) and decreased glutathione-S-transferase activity in hepatic tissue, compared to control group. Enhanced lipid peroxidation (LPO) levels along with decreased reduced glutathione (GSH) level were also observed in DMBA group, compared to control group. AAILE co-treatment (200 mg/kg body weight, po, thrice a week) for 8 weeks followed by DMBA injection showed significant improvement in hepatic status, as revealed by normalization of 99mTc-mebrofenin clearance rate, decreased ALP and ALT levels, reduced genotoxicity in terms of micronuclei score and apoptotic index. Levels of LPO were significantly decreased along with increased hepatic GST and GSH levels in AAILE + DMBA group, compared to DMBA group. However, no significant change was observed in hepatic CYP and Cyt b5 levels, compared to DMBA group. The results indicated that AAILE effectively ameliorated DMBA-induced hepatotoxicity.

Modulatory effects of Azadirachta indica leaf extract on cutaneous and hepatic biochemical status during promotion phase of DMBA/TPA-induced skin tumorigenesis in mice.

The modulation in biochemical status of skin and hepatic tissue at the time point of commencement of promotion stage of skin carcinogenesis in mice and its intervention with aqueous Azadirachta indica leaf extract (AAILE) were investigated. 7,12-Dimethylbenz(a)anthracene (DMBA, 500 nmol/100 ul of acetone) was applied topically for 2 weeks (twice weekly), followed by phorbol-12-myristate-13-acetate (TPA, 1.7 nmol/100 ul) twice weekly for 6 weeks on the depilated skin of mice and AAILE was administered orally at a dose level of 300 mg/kg body wt thrice a week for 10 weeks. DMBA/TPA treatment upregulated the phase I enzymes in skin and hepatic tissue, as revealed by the increased cytochrome P450 (CYP) and cytochrome b5 (cyt b5) levels and aryl hydrocarbon hydroxylase (AHH) activity when compared to the control group and differentially modulated the activities of phase II enzymes like glutathione-s-transferase (GST), DT-diaphorase (DTD) and uridine diphosphate glucuronosyltransferase (UDP-GT). AAILE treatment decreased the DMBA/TPA-induced increase in cutaneous CYP level and enhanced the DTD and UDP-GT activities when compared with DMBA/TPA group. In the hepatic tissue of AAILE + DMBA/TPA group, an increase in UDP-GT activity was observed when compared to DMBA/TPA group. DMBA/TPA treatment did not alter the skin lipid peroxidation (LPO) level when compared to control group, however, in the animals that received AAILE treatment along with DMBA/TPA, a significant increase in LPO was observed when compared to control group. This was associated with a decrease in cutaneous reduced glutathione (GSH) level of AAILE + DMBA/TPA group. Enhanced LPO level was observed in the hepatic tissue of DMBA/TPA and AAILE + DMBA/TPA groups when compared to control group. However, no alteration was observed in their hepatic GSH levels. The micronuclei score in hepatic tissue did not exhibit significant inter-group differences. The results of the present study suggest that apart from skin, liver may be affected during DMBA/TPA-induced skin tumorigenesis. AAILE treatment has the ability to modulate these changes potentially influencing the process of tumor formation. These findings seem to be important to carcinogenesis and its intervention with anti-cancer agents.

Properties of catechol 1, 2 dioxygenase in the cell free extract and immobilized extract of Mycobacterium fortuitum

Polycyclic aromatic hydrocarbons (PAH) are carcinogenic compounds which contaminate water and soil, and the enzymes can be used for bioremediation of these environments. This study aimed to evaluate some environmental conditions that affect the production and activity of the catechol 1,2-dioxygenase (C12O) by Mycobacterium fortuitum in the cell free and immobilized extract in sodium alginate. The bacterium was grown in mineral medium and LB broth containing 250 mg L-1 of anthracene (PAH). The optimum conditions of pH (4.0-9.0), temperature (5-70 ºC), reaction time (10-90 min) and the effect of ions in the enzyme activity were determined. The Mycobacterium cultivated in LB shown higher growth and the C12O activity was two-fold higher to that in the mineral medium. To both extracts the highest enzyme activity was at pH 8.0, however, the immobilized extract promoted the increase in the C12O activity in a pH range between 4.0 and 8.5. The immobilized extract increased the enzymatic activity time and showed the highest C12O activity at 45 ºC, 20 ºC higher than the greatest temperature in the cell free extract. The enzyme activity in both extracts was stimulated by Fe3+, Hg2+ and Mn2+ and inhibited by NH4+ and Cu2+, but the immobilization protected the enzyme against the deleterious effects of K+ and Mg2+ in tested concentrations. The catechol 1,2-dioxygenase of Mycobacterium fortuitum in the immobilized extract has greater stability to the variations of pH, temperature and reaction time, and show higher activity in presence of ions, comparing to the cell free extract

Effect of Paclitaxel and Paclitaxel-Di Allyl Sulfide Combination on 7, 12 Dimethyl Benz (A) Anthracene (DMBA) Induced Skin Cancer in Wistar Rats: A Comparative Study.

Cancer is a cellular tumor that unlike benign tumor can metastasize and invade the surrounding tissues. In the present study the anticancer effect of paclitaxel was evaluated on 7, 12 Di Methyl Benz (a) Anthracene induced skin cancer in wistar rats and results were compared with normal, paclitaxel and paclitaxel-Di allyl sulfide combined alternative chemotherapy. By analyzing the various biochemical parameters (lipid Profile and lipid metabolizing enzymes) and Marker enzymes (squamous cell antigen). Skin cancer was induced in rats by 7, 12 Di methyl benz(a) anthracene (DMBA) at the dosage of 5 μg was dissolved in 100μl and administered into experimental animals for 28 weeks. In this study, we demonstrated that combination of paclitaxel and Di allyl sulfide protects the rats from a lethal dose of DMBA for 30 days. Total cholesterol (TC), free cholesterol (FC), phospholipids (PL) and triglycerides (TG) were found to be significantly increased whereas ester cholesterol (EC) and free fatty acids (FFA) were significantly decreased when compared with cancer bearing group II animals. From the present study, the effect of Paclitaxel- Di allyl sulfide combination proved to be a more significant chemotherapeutic agent against DMBA induced skin cancer in wistar rats compared to that of paclitaxel by analyzing the lipid profile and lipid metabolizing enzymes and marker enzymes.

Chemopreventive potential of geraniol in 7,12-dimethylbenz(a) anthracene (DMBA) induced skin carcinogenesis in Swiss albino mice.

The present study investigated the chemopreventive potential of geraniol, an acyclic monoterpene alcohol, by monitoring the tumor incidence and analyzing the status of phase II detoxification agents, lipid peroxidation by products and antioxidants in 7,12-dimethylbenz(a)anthracene (DMBA) induced mouse skin carcinogenesis. Skin tumor was developed by painting DMBA (25 μg in 0.1 ml acetone mouse-1) in the shaved back of the mice, twice weekly for 8 weeks. We noticed 100% skin tumor formation in mice treated with DMBA alone. The status of phase II detoxification agents and antioxidants were decreased where as lipid peroxidation by products were increased in tumor bearing mice. Oral administration of geraniol at a dose of 250 mg kg-1 body weight significantly prevented the tumor formation as well as brought back the status of phase II detoxification agents, lipid peroxidation by products and antioxidants to near normal range in DMBA treated mice. Present results suggest that geraniol might have inhibited abnormal cell proliferation occurring in skin carcinogenesis by modulating the activities of phase II detoxification agents and through its free radical scavenging potential.