Apoptotic and autophagic death union by Thuja occidentalis homeopathic drug in cervical cancer cells with thujone as the bioactive principle / 中西医结合学报

OBJECTIVE@#"Multi-targeting" drugs can prove fruitful to combat drug-resistance of multifactorial disease-cervical cancer. This study envisioned to reveal if Thuja homeopathic mother tincture (MT) and its bioactive component could combat human papillomavirus (HPV)-16-infected SiHa cervical cancer cells since it is globally acclaimed for HPV-mediated warts.@*METHODS@#Thuja MT was studied for its antiproliferative and antimigratory properties in SiHa cells followed by microscopic determination of reactive oxygen species (ROS) generation by 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) staining and loss in mitochondrial membrane potential (MtMP) by rhodamine 123 (Rh123) staining. Apoptosis and autophagy inductions were studied by acridine orange/ethidium bromide (AO/EB) staining and immunoblot analyses of marker proteins. The bioactive component of Thuja MT detected by gas chromatography-mass spectrometry was studied for antiproliferative and antimigratory properties along with in silico prediction of its cellular targets by molecular docking and oral drug forming competency.@*RESULTS@#Thuja MT showed significant antiproliferative and antimigratory potential in SiHa cells at a 50% inhibitory concentration (IC50) of 17.3 µL/mL. An increase in DCFDA fluorescence and loss in Rh123 fluorescence prove that Thuja MT acted through the burst of ROS and loss in MtMP respectively. AO/EB-stained cells under the microscope and immunoblot analyses supported Thuja-induced cellular demise via dual pathways-apoptosis and autophagy. Immunoblots showed cleavage of caspase-3 and poly(adenosine diphosphate-ribose) polymerase-1 (PARP-1) along with upregulation of Beclin-1, microtubule-associated protein 1 light chain 3B (LC3B)-II, and p62 proteins. Hence, the apoptotic cascade followed a caspase-3-dependent pathway supported by PARP-1 cleavage, while autophagic death was Beclin-1-dependent and mediated by accumulation of LC3BII and p62 proteins. Thujone, detected as the bioactive principle of Thuja MT, showed greater anti-proliferative and anti-migratory potential at an IC50 of 77 µg/mL, along with excellent oral drug competency with the ability for gastrointestinal absorption and blood-brain-barrier permeation with nil toxicity. Molecular docking depicted thujone with the strongest affinity for mammalian target of rapamycin, phosphoinositide 3-kinase, and protein kinase B followed by B-cell lymphoma 2, murine double minute 2 and adenosine monophosphate-activated protein kinase, which might act as upstream triggers of apoptotic-autophagic crosstalk.@*CONCLUSION@#Robust "multi-targeting" anticancer potential of Thuja drug and thujone for HPV-infected cervical cancer ascertained its therapeutic efficacy for HPV infections.

Tratamiento Homeopático de la Verruga Plantar. Caso Clínico Tratado en una Farmacia Comunitaria / Homeopathic Wart Treatment Plant. Clinical Case Treated in a Community Pharmacy

Las verrugas plantares son un tipo de infección causadas por el virus del papiloma humano (VPH). En muchos de los casos son muy dolorosas y difíciles de tratar; aunque generalmente desaparecen con tratamiento en unos meses, en otros casos persisten durante años. Se presenta el caso de un varón de 13 años que acudió a la oficina de farmacia tras el fracaso de un prolongado tratamiento convencional para una verruga plantar en el talón. Existen antecedentes documentados y experiencias favorables en el tratamiento homeopático de las verrugas plantares y de su utilización en farmacia. Se le pautaron Thuja occidentalis 15CH (10 gránulos semanalmente), Dulcamara 9CH (5-0-5), Antimonium crudum 15CH (5-0-5) y Nitricum acidum 15CH (5-0-5) durante 1 mes. Se observó una mejoría clínica importante al cabo de sólo 1 mes exclusivamente con tratamiento homeopático, que puede suponer una posibilidad de tratamiento eficaz para las verrugas plantares, incluso tras el fracaso de otras intervenciones convencionales.(AU)

Plantar warts are a type of infection caused by the human papillomavirus. In many cases they are very painful and difficult to treat, they usually disappear in a few months with the treatment but in several cases they will persist for years. The case of a 13 years old male is presented, who came to the pharmacy after a long and unsuccessful conventional treatment of a plantar wart on the heel of the left foot. The plantar warts have been successfully treated by homeopathic medicines in the bibliography and according to our own experience. The chosen medicines were Thuja occidentalis 15CH (10 granules weekly), Dulcamara 9CH 5 (5-0-5), Antimonium crudum 15CH (5-0-5) and Nitricum acidum 15CH (5-0-5). An important improvement was observed after 1 month of treatment. The homeopathic treatment can be an effective treatment option for plantar warts, including those where other conventional interventions have not succeeded. (AU)

Antimicrobial activity of Cannabis sativa, Thuja orientalis and Psidium guajava leaf extracts against methicillin-resistant Staphylococcus aureus / 中西医结合学报

<p><b>OBJECTIVE</b>This study examined the antimicrobial activity of Cannabis sativa, Thuja orientalis and Psidium guajava against methicillin-resistant Staphylococcus aureus (MRSA) and used a standardized purification protocol to determine the presence and abundance of bioactive compounds in the leaf extracts.</p><p><b>METHODS</b>In vitro antimicrobial activities of the ethanolic extracts of C. sativa, T. orientalis and P. guajava were tested against MRSA. The presence of bioactive molecules in these three leaves was evaluated using biochemical assays and high-performance thin-layer chromatography (HPTLC).</p><p><b>RESULTS</b>Resistance to methicillin, penicillin, oxacillin and cefoxitin was observed in each of the clinical and nonclinical MRSA isolates. However, they were still vulnerable to vancomycin. Used individually, the 50% extract of each plant leaf inhibited MRSA growth. A profound synergism was observed when C. sativa was used in combination with T. orientalis (1:1) and when P. guajava was used in combination with T. orientalis (1:1). This was shown by larger zones of inhibition. This synergism was probably due to the combined inhibitory effect of phenolics present in the leaf extracts (i.e., quercetin and gallic acid) and catechin, as detected by HPTLC.</p><p><b>CONCLUSION</b>The leaf extracts of C. sativa, T. orientalis and P. guajava had potential for the control of both hospital- and community-acquired MRSA. Moreover, the inhibitory effect was enhanced when extracts were used in combination.</p>

Doseamento de flavonoides totais das partes aéreas de Thuja occidentalis Linn. (Cupressaceae) / Assay of total flavonoids from aerial parts of Thuja occidentalis Linn. (Cupressaceae)

Thuja occidentalis é largamente utilizada na forma de tinturas, sendo os flavonoides metabólitos de interesse da espécie, visto as distintas atividades farmacológicas citadas na literatura. Contudo, não há descrito método para a quantificação desses, sendo objetivo desse trabalho desenvolver um método colorimétrico direto utilizando cloreto de alumínio (AlCl3) por espectrofotometria UV, a fim de proporciona a quantificação de flavonoides totais, expresso em rutina. Para tal, avaliaram-se os seguintes parâmetros: concentração da solução hidroalcoólica (SH) (20, 40 e 70%); interferentes lipofílicos; volume da solução de AlCl3 2,5% (1, 2 e 3 mL) e proporção de droga vegetal (0,5, 1, 1,5 g). Adicionalmente, procedeuse a validação. A partir da avaliação dos interferentes lipofílicos, selecionou-se a SH 40% utilizando 1 g da droga vegetal. A cinética de complexação, demonstrou que a formação do complexo flavonoides-Al+3 é lenta, sendo o tempo de leitura ideal de 30 min no λ=403 nm, após a adição de 2 mL de AlCl3 2,5%. A rutina foi utilizada a conversão de absorvância em teor de flavonoides totais, uma vez que a espécie possui a presença majoritária de derivados quercetínicos do grupo flavonol na sua forma glicosilada. O método atendeu a todos os requisitos indicados para validação de uma matriz vegetal, consideração os fatores inerentes da espécie, possibilitando sua utilização na prática laboratorial.

Homoeopathic treatment of vitiligo: an open observational pilot study

Background: Vitiligo is a common hypopigmentation disorder with significant psychological impact. An evaluation of homeopathic treatment was performed in individuals with vitiligo in a hospital outpatient clinic in West Bengal, India. Methods: Thirty participants (median age 27 years old, 57% female) were recruited for a prospective open-label pilot study and treated with individualized homeopathic medicines for 6 months. Efficacy was assessed after 3 months and 6 months using Vitiligo Area Scoring Index (VASI) score, Vitiligo European Task Force (VETF) score, and Dermatological Life Quality Index (DLQI) score, which are validated outcome measures evaluating the area, intensity, spread of depigmentation of vitiligo lesions, and quality of life (QoL). Results: A total of 27 participants completed the trial; 3 dropped out. After 6 months of treatment, the median VASI total score improved significantly by 0.1 units (p=0.003), from 0.8 (0.5, 1.5) to 0.7 (0.3, 0.8) on a scale from 0 (no depigmentation) to 100 (completely depigmented). Similarly, the VETF median score improved by 2 units (p=0.0001) from 1 (0, 1) to –1 (–1, 0) and the staging score changed from 1 (1, 2) to 1 (0, 1), p=0.002. The total DLQI median score exhibited significant reduction from 21 (17, 22) to 13.6 (10, 17), change 7.4 (p=0.0001), as also did its components. Conclusions: Individualized homeopathic treatment associated with significant improvement of VASI, VETF and DLQI scores. The extent to which the observed effects were due to placebo needs clarification in future randomized double-blind clinical studies preceded by feasibility studies.

Introdução: Vitiligo é uma desordem de hipopigmentação de ocorrência comum, com impacto psicológico significativo. Foi avaliado o tratamento homeopático de indivíduos com vitiligo no ambulatório de um hospital em Bengala Ocidental, Índia. Métodos: Trinta participantes (idade mediana 27 anos, sendo 57% do gênero feminino) foram recrutados para o presente estudo piloto prospectivo aberto e tratados com medicamentos homeopáticos individualizados por 6 meses. A eficácia do tratamento foi avaliada aos 3 e 6 meses mediante Vitiligo Area Scoring Index (VASI) score, Vitiligo European Task Force (VETF), e Dermatological Life Quality Index (DLQI), que são medidas validadas de desfecho correspondendo à área, intensidade e disseminação da depigmentação causada por vitiligo, assim como a qualidade de vida (QV). Resultados: Um total de 27 participantes completou o estudo, sendo que 3 abandonaram. Depois de 6 meses de tratamento, a mediana do escore total do VASI apresentou melhora significativa de 0,1 unidade (p=0,003) passando de 0,8 (0,5, 1,5) para 0,7 (0,3, 0,8), numa escala de 0 (sem depigmentação) a 100 (depigmentação completa). De modo similar, a mediana do escore VETF de disseminação melhorou em 2 unidades (p=0,0001) de 1 (0, 1) a –1 (–1, 0), e o escore de estadiamento variou de 1 (1, 2) para 1 (0, 1), p=0,002. O escore total mediano de DLQI diminuiu de 21 (17, 22) a 13,6 (10, 17), com uma variação de 7,4 (p=0,0001); variação também foi constatada nos componentes de DLQI. Conclusões: O tratamento homeopático individualizado se associou com melhora significativa nos escores de VASI, VETF e DLQI. A parte dos efeitos possivelmente devida a efeito placebo precisa ser determinada em futuros estudos clínicos randomizados e duplo cego, após a realização de estudos de factibilidade.

Homeopathic Thuja 30C ameliorates benzo(a)pyrene-induced DNA damage, stress and viability of perfused lung cells of mice in vitro / 中西医结合学报

<p><b>OBJECTIVE</b>To examine if the ultra-highly diluted homeopathic remedy Thuja 30C can ameliorate benzo(a)pyrene (BaP)-induced DNA damage, stress and viability of perfused lung cells of Swiss albino mice in vitro.</p><p><b>METHODS</b>Perfused normal lung cells from mice were cultured in 5% Roswell Park Memorial Institute medium and exposed to BaP, a potent carcinogen, at the half maximal inhibitory concentration dose (2.2 μmol/L) for 24 h. Thereafter, the intoxicated cells were either treated with Thuja 30C (used against tumor or cancer) or its vehicle media, succussed alcohol 30C. Relevant parameters of study involving reactive oxygen species (ROS) accumulation, total glutathione (GSH) content, and generations of heat shock protein (hsp)-90 were measured; the cell viability and other test parameters were measured after treatment with either Thuja 30C or its vehicle media. Circular dichroism spectroscopy was performed to examine if Thuja 30C directly interacted with calf thymus DNA as target. For ascertaining if DNA damaged by BaP could be partially repaired and restituted by the remedy, 4',6-diamidino-2-phenylindole staining was performed.</p><p><b>RESULTS</b>Thuja 30C increased cell viability of BaP-intoxicated cells significantly, as compared to drug-untreated or drug-vehicle control. A minimal dose of Thuja 30C significantly inhibited BaP-induced stress level, by down-regulating ROS and hsp-90, and increasing GSH content. Thuja 30C itself had no DNA-damaging effect, and no direct drug-DNA interaction. However, it showed quite striking ability to repair DNA damage caused by BaP.</p><p><b>CONCLUSION</b>Thuja 30C ameliorates BaP-induced toxicity, stress and DNA damage in perfused lung cells of mice and it apparently has no effect on normal lung cells.</p>

The Revised Edition of Korean Calendar for Allergenic Pollens

The old calendar of pollens did not reflect current pollen distribution and concentrations that can be influenced by changes of weather and environment of each region in South Korea. A new pollen calendar of allergenic pollens was made based on the data on pollen concentrations obtained in eight regions nationwide between 1997 and 2009. The distribution of pollen was assessed every day at 8 areas (Seoul, Guri, Busan, Daegu, Jeonju, Kwangju, Kangneung, and Jeju) for 12 years between July 1, 1997 and June 30, 2009. Pollens were collected by using Burkard 7-day sampler (Burkard Manufacturing Co Ltd, UK). Pollens which were stained with Calberla's fuchsin staining solution were identified and counted. Pine became the highest pollen in May, and the pollen concentrations of oak and birch also became high. Ragweed appeared in the middle of August and showed the highest pollen concentration in the middles of September. Japanese hop showed a high concentration between the middle of August and the end of September, and mugwort appeared in the middles of August and its concentration increased up until early September. In Kangneung, birch appeared earlier, pine showed a higher pollen concentration than in the other areas. In Daegu, Oriental thuja and alder produced a large concentration of pollens. Pine produced a large concentration of pollens between the middle of April and the end of May. Weeds showed higher concentrations in September and mugwort appeared earlier than ragweed. In Busan the time of flowering is relatively early, and alder and Oriental thuja appeared earliest among all areas. In Kwangju, Oriental thuja and hazelnut appeared in early February. Japanese cedar showed the highest pollen concentration in March in Jeju. In conclusion, update information on pollen calendar in South Korea should be provided for allergic patients through the website to manage and prevent the pollinosis.

Effect of homeopathic medicines on transplanted tumors in mice.

Ultra low doses used in homeopathic medicines are reported to have healing potential for various diseases but their action remains controversial. In this study we have investigated the antitumour and antimetastatic activity of selected homeopathic medicines against transplanted tumours in mice. It was found that Ruta graveolens 200c and Hydrastis canadensis 200c significantly increased the lifespan of Ehrlich Ascites Carcinoma and Dalton's Lymphoma Ascites induced tumour-bearing animals by 49.7%, and 69.4% respectively. Moreover there was 95.6% and 95.8% reduction of solid tumour volume in Ruta 200c and Hydrastis 200c treated animals on the 31st day after tumour inoculation. Hydrastis 1M given orally significantly inhibited the growth of developed solid tumours produced by DLA cells and increased the lifespan of tumour bearing animals. Some 9 out of 15 animals with developed tumors were completely tumour free after treatment with Hydrastis 1M. Significant anti-metastatic activity was also found in B16F-10 melanoma-bearing animals treated with Thuja1M, Hydrastis 1M and Lycopodium1M. This was evident from the inhibition of lung tumour nodule formation, morphological and histopathological analysis of lung and decreased levels of gamma-GT in serum, a cellular marker of proliferation. These findings support that homeopathic preparations of Ruta and Hydrastis have significant antitumour activity. The mechanism of action of these medicines is not known at present.

Ineficiência da Thuja occidentalis no tratamento dos poxvirus aviários / Thuja occidentalis ineficiency in avian poxviruses treatment

Medicações à base de Tuia (Thuja occidentalis) são comuns em tratamentos homeopáticos e fitoterápicos, na medicina humana e veterinária. Suas propriedades imunoestimulantes e antivirais são descritas e sua utilização empírica no tratamento das poxviroses aviárias é recomendada por criadores e veterinários. Para avaliar o potencial terapêutico da Tuia sobre estas infecções, dois grupos (controle e teste) de dez aves jovens foram inoculados por escarificação, com uma amostra de campo de poxvírus aviário. As aves inoculadas foram examinadas diariamente, quanto ao início e evolução das lesões, registrando-se o número e tamanho destas, até a cicatrização. Após o surgimento das lesões, o grupo teste recebeu a tintura alcoólica diluída na água de consumo. A análise experimental indicou que, nas condições empregadas, a utilização da Tuia não favoreceu a regressão ou restrição no desenvolvimento das lesões de pele promovidas pela infecção experimental.

Larvicidal potential of Nerium indicum and Thuja oriertelis extracts against malaria and Japanese encephalitis vector.

Ethanolic and acetone extracts of Nerium indicum and Thuja orientelis have been studied against III instar larvae of Anopheles stephensi and Culex quinquefasciatus. Ethanolic extract of N. indicum is found more effective than its acetone extract against anopheline larvae with LC50 values of 185.99 and 148.05 ppm for former and 229.28 and 149.43 ppm for the later after 24 and 48 hrs of exposure. The acetone extract with LC50 values of 209.00 and 155.97 ppm is more effective in case of culicine larvae than its ethanolic extract with LC50 494.07 and 194.49 ppm after 24 and 48 hours of treatment. Ethanolic extract of T. orientelis is more effective against both the larval species with LC50 values of 13.10 and 9.02 ppm after 24 and 48 hours for anopheline and 22.74 and 16.72 ppm against culicine larvae. The acetone extract showed LC50 values of 200.87 and 127.53 ppm against anopheline and 69.03 and 51.14 ppm against culicine larvae. Thus ethanolic extract of T. orientelis is an ideal potential larvicide for both types of mosquito larvae.

Growth Inhibitory Effects of Various Herbal Extracts on Metronidazole Resistant Strain of Trichomonas vaginalis

BACKGROUND: Metronidazole has been known as the most effective drug for treatment of Trichomonas vaginalis-related diseases. However, it has been reported that metronidazole has adverse effects and incidence of metronidazole-resistant T. vaginalis (CDC085) has increased. Development of new drug, which is effective against metronidazole-resistant T. vaginalis and showing no adverse effects, has been required. METHODS: The purpose of this study was to investigate effects of various extracts from herbs such as Quisqualis indica, Gleditsia sinensis, Prunus armeniaca, Morus alba, Platycodon grandiflorum, Ailanthus altissima, Stemona japonica, Biota orientalis, Dryobalanops aromatica, and Cimicifuga heracleifolia on metronidazole resistant strain of T. vaginalis in vitro (CDC085). RESULTS: Anti-Trichomonas activities were observed in T. vaginalis treated with G. sinensis, P. armeniaca, and P. grandiflorum on the growth and fine structure of metronidazole resistant strain of T. vaginalis. Of the three standard extracts that showed the most effective anti-trichomonas activity, G. sinensis was the most effective. The inhibitory effects of fraction extracts of this drug were shown on the growth of T. vaginalis. The fine structure of the cytoplasm was changed after application of G. sinensis extract. The number of polyribosome and hydrogenosome decreased whereas the number of food vacuole and vacuole in the cytoplasm increased, compared with that of untreated control group. CONCLUSION: The results of our study indicate that G. sinensis may induce the inhibition of cell multiplication as well as impairment of protein synthesis of metronidazole resistant strain of T. vaginalis in vitro.

Growth Inhibitory Effects of Various Herbal Extracts on Metronidazole Resistant Strain of Trichomonas vaginalis

BACKGROUND: Metronidazole has been known as the most effective drug for treatment of Trichomonas vaginalis-related diseases. However, it has been reported that metronidazole has adverse effects and incidence of metronidazole-resistant T. vaginalis (CDC085) has increased. Development of new drug, which is effective against metronidazole-resistant T. vaginalis and showing no adverse effects, has been required. METHODS: The purpose of this study was to investigate effects of various extracts from herbs such as Quisqualis indica, Gleditsia sinensis, Prunus armeniaca, Morus alba, Platycodon grandiflorum, Ailanthus altissima, Stemona japonica, Biota orientalis, Dryobalanops aromatica, and Cimicifuga heracleifolia on metronidazole resistant strain of T. vaginalis in vitro (CDC085). RESULTS: Anti-Trichomonas activities were observed in T. vaginalis treated with G. sinensis, P. armeniaca, and P. grandiflorum on the growth and fine structure of metronidazole resistant strain of T. vaginalis. Of the three standard extracts that showed the most effective anti-trichomonas activity, G. sinensis was the most effective. The inhibitory effects of fraction extracts of this drug were shown on the growth of T. vaginalis. The fine structure of the cytoplasm was changed after application of G. sinensis extract. The number of polyribosome and hydrogenosome decreased whereas the number of food vacuole and vacuole in the cytoplasm increased, compared with that of untreated control group. CONCLUSION: The results of our study indicate that G. sinensis may induce the inhibition of cell multiplication as well as impairment of protein synthesis of metronidazole resistant strain of T. vaginalis in vitro.