Active constituents and mechanisms of Respiratory Detox Shot, a traditional Chinese medicine prescription, for COVID-19 control and prevention: Network-molecular docking-LC-MS analysis / 中西医结合学报

OBJECTIVE@#Lung-toxin Dispelling Formula No. 1, referred to as Respiratory Detox Shot (RDS), was developed based on a classical prescription of traditional Chinese medicine (TCM) and the theoretical understanding of herbal properties within TCM. Therapeutic benefits of using RDS for both disease control and prevention, in the effort to contain the coronavirus disease 2019 (COVID-19), have been shown. However, the biochemically active constituents of RDS and their mechanisms of action are still unclear. The goal of the present study is to clarify the material foundation and action mechanism of RDS.@*METHODS@#To conduct an analysis of RDS, an integrative analytical platform was constructed, including target prediction, protein-protein interaction (PPI) network, and cluster analysis; further, the hub genes involved in the disease-related pathways were identified, and the their corresponding compounds were used for in vitro validation of molecular docking predictions. The presence of these validated compounds was also measured in samples of the RDS formula to quantify the abundance of the biochemically active constituents. In our network pharmacological study, a total of 26 bioinformatic programs and databases were used, and six networks, covering the entire Zang-fu viscera, were constructed to comprehensively analyze the intricate connections among the compounds-targets-disease pathways-meridians of RDS.@*RESULTS@#For all 1071 known chemical constituents of the nine ingredients in RDS, identified from established TCM databases, 157 passed drug-likeness screening and led to 339 predicted targets in the constituent-target network. Forty-two hub genes with core regulatory effects were extracted from the PPI network, and 134 compounds and 29 crucial disease pathways were implicated in the target-constituent-disease network. Twelve disease pathways attributed to the Lung-Large Intestine meridians, with six and five attributed to the Kidney-Urinary Bladder and Stomach-Spleen meridians, respectively. One-hundred and eighteen candidate constituents showed a high binding affinity with SARS-coronavirus-2 3-chymotrypsin-like protease (3CL), as indicated by molecular docking using computational pattern recognition. The in vitro activity of 22 chemical constituents of RDS was validated using the 3CL inhibition assay. Finally, using liquid chromatography mass spectrometry in data-independent analysis mode, the presence of seven out of these 22 constituents was confirmed and validated in an aqueous decoction of RDS, using reference standards in both non-targeted and targeted approaches.@*CONCLUSION@#RDS acts primarily in the Lung-Large Intestine, Kidney-Urinary Bladder and Stomach-Spleen meridians, with other Zang-fu viscera strategically covered by all nine ingredients. In the context of TCM meridian theory, the multiple components and targets of RDS contribute to RDS's dual effects of health-strengthening and pathogen-eliminating. This results in general therapeutic effects for early COVID-19 control and prevention.

Expression of ?-smooth muscle actin in benign and malignant salivary gland tumors: An immunohistochemical study

Background and Objectives: Myoepithelial cells (ME) are known to contribute in the patterning of salivary gland neoplasms (SGN) and possess cytoplasmic smooth muscle actin (SMA) revealed by alpha SMA (?-SMA). The present study aimed to assess the expression of ?-SMA in selected benign and malignant SGN (pleomorphic adenoma printarticle.asp?issn=0377-4929;year=2018;volume=61;issue=4;spage=479;epage=484;aulast=Ravi, mucoepidermoid carcinoma (MEC), adenoid cystic carcinoma (ACC), and polymorphous low-grade adenocarcinoma (PLGA). Materials and Methods: The intensity and pattern of expression of ?-SMA were studied in 25 cases of SGN's ACC (n = 7), MEC (n = 8), PA (n = 8), and PLGA (n = 2), and correlated with the histological patterns. Results: Maximum expression of ?-SMA in the epithelial compartment was seen in ACC, followed by PA, whereas MEC and PLGA showed completely negative staining. The connective tissue expression was mild in ACC and MEC. The myxoid stroma of PA with “melting” pattern was weakly positive for ?-SMA. The stroma in PLGA showed complete negativity. In ACC, ?-SMA-positive cells were lining the cribriform spaces, small islands, and dispersed within large islands. Small nests showed complete positivity for ?-SMA. Interpretation and Conclusion: In ACC, ?-SMA expression supports the involvement of ME in epithelial organization explaining the histological patterns seen. In PA, the expression correlates with the predominantly secretory nature of ME. The absence of epithelial positivity in MEC and PLGA suggest that ME has less role to play in their histogenesis. The weak stromal positivity observed in MEC and ACC may be attributed to the positive immunoreactivity of myofibroblasts playing a role in modulating the course of SGN's.

Cassia alata leaf extract induces cytotoxicity in A549 lung cancer cells via a mechanism that is caspase 8 dependent / El extracto de la hoja de cassia alata induce citotoxicidad en las células A549 del cáncer pulmonar humano a través de un mecanismo dependiente de la caspasa 8

OBJECTIVE: To evaluate the cytotoxic effect of a hexane extract of Cassia alata leaves in A549 lung cancer cells. METHOD: Parental A549 lung cancer cells were exposed to various concentrations (100"180 µg/ml) of Cassia alata leaf extract for 24 hours. Following treatment, the cells were evaluated using the 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay to determine the cytotoxic effect of the extract. Caspase 8, 3 and 9 negative A549 cells were also prepared using lentiviral based shRNA knockdown of the caspase 8, 3 and 9 genes, respectively. The cytotoxic effect of Cassia alata leaf extract was then evaluated in these knockdown cells using the MTT assay. Chemical analysis was performed on the extract using high performance liquid chromatography (HPLC). RESULTS: Cassia alata extract was cytotoxic in parental and caspase-9 negative, but not caspase 3 and 8 negative A549 cells. The IC50 values were 143 µg/ml and 145 µg/ml in parental and caspase 9 negative A549 cells respectively. The flavanoid kaempferol was identified as a constituent of Cassia alata leaf extract. CONCLUSIONS: Cassia alata produces cytotoxicity in A549 cancer cells that is mediated by caspase 8 activation. This effect may be attributable to kaempferol.

OBJETIVO: Evaluar el efecto citotóxico de un extracto de hexano de hojas de Cassia alata en las células A549 del cáncer pulmonar. MÉTODO: Células A549 parentales del cáncer pulmonar fueron expuestas a varias concentraciones (100-180 µg/ml) de un extracto de la hoja de Cassia alatadurante 24 horas. Tras el tratamiento, las células fueron evaluadas usando el ensayo de bromuro de 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazolio (MTT) a fin de determinar el efecto citotóxico del extracto. También se prepararon células A549 negativas caspasa 8, 3 y 9 mediante silenciamiento génico vía ARN (shRNA knockdown) de los genes de las caspasas 8, 3 y 9 respectivamente, sobre la base de la inserción de vectores lentivirales. Entonces, usando un ensayo MTT se procedió a evaluar el efecto citotóxico del extracto de hojas de Cassia alataen éstas células genéticamente modificadas. Se realizó un análisis químico del extracto utilizando cromatografía líquida de alta eficacia. (HPLC). RESULTADOS: El extracto de Cassia alata resultó ser citotóxico en las células A549 negativas parentales y caspasa 9, pero no en las negativas caspasa 3 y 8. Los valores de IC50 fueron 143 µg/ml y 145 µg/ml en las células A549 negativas parentales y caspasa 9 respectivamente. El flavonol kaempferol fue identificado como un constituyente del extracto de las hojas de Cassia alata. CONCLUSIONES: La Cassia alata produce citotoxicidad en las células cancerosas A549, mediada por la activación de la caspasa 8. Este efecto puede ser atribuido al kaempferol.

Anti-inflammatory activities of cassia alata leaf extract in complete freund's adjuvant arthritis in rats / Actividades anti-inflamatorias del extracto de hojas de cassia alata en artritis inducida en ratas mediante adyuvante completo de freund

OBJECTIVE: To investigate the anti-inflammatory effects of a hexane extract of Cassia alata leaves in complete Freund's adjuvant (CFA) arthritis in rats. METHOD: A hexane extract of Cassia alata leaves was administered by oral gavage to CFA arthritic rats (500 mg/kg, n = 6). Controls received corn oil (2 ml, n = 6). The CFA arthritic model was induced by the injection of 0.5 ml (CFA) into the synovial cavity of the right knee joint of the hind leg of rats. The ability of the plant extract to reduce swelling as a sign of arthritic inflammation was assessed by obtaining the circumference of the knee joint before and for twenty eight days post arthritis induction. Reduction of leukocyte infiltration into the blood and synovial cavity of the arthritic rats were assessed using automated counting and Wrights method. Protection against cartilage erosion was also assessed histologically. RESULTS: Cassia alata extract significantly (p = 0.0032) reduced knee circumference (swelling) in the CFA arthritic rats. Total and differential leukocyte counts in both blood and synovial fluid from Cassia alata treated animals were significantly (p < 0.05) lower than in control animals. Protective effects against cartilage degradation on the femoral head of the knee joint were observed in Cassia alata treated animals, as normal cartilage structure and chondrocyte arrangement were maintained. CONCLUSIONS: The results suggest that Cassia alata exhibits anti-inflammatory activities that should be further examined and potentially exploited for anti-arthritic therapies.

OBJETIVO: Investigar los efectos anti-inflamatorios del extracto de hexano de hojas de Cassia alata en artritis inducida por adyuvante completo de Freund (CFA) en ratas. MÉTODO: Un extracto de hexano de hojas de Cassia alata fue administrado por gavage oral a ratas artríticas por CFA (500 mg/kg, n = 6). Los controles recibieron aceite de maíz (2 ml, n = 6). El modelo artrítico de CFA fue inducido inyectando 0.5 ml (CFA) en la cavidad sinovial de la rótula derecha de la pata trasera de las ratas. La capacidad del extracto de la planta en cuanto a reducir la inflamación como signo de la inflamación artrítica, fue evaluada obteniendo la circunferencia de la rótula antes y durante veintiocho días posterior a la inducción de la artritis. La reducción de la infiltración de leucocitos en la sangre y la cavidad sinovial de las ratas artríticas fue evaluada usando el conteo automatizado y el método de Wright. También se evaluó histológicamente la protección contra la erosión del cartílago. RESULTADOS: El extracto de Cassia alata redujo significativamente (p = 0.0032) la circunferencia de la rodilla (inflamación) en las ratas artríticas por CFA. Los conteos totales y diferenciales de leucocitos tanto en la sangre como en el líquido sinovial de los animales tratados con Cassia alata fueron significativamente (p < 0.05) más bajos en los animales del control. Los efectos protectores contra la degradación del cartílago en la cabeza femoral de la rótula fueron observados en los animales tratados con Cassia alata, ya que se mantuvieron la estructura normal del cartílago y las disposición de los condrocitos. CONCLUSIONES: Los resultados sugieren que la Cassia alata exhibe propiedades anti-inflamatorias que deben ser examinadas ulteriormente y explotadas potencialmente para las terapias anti-artríticas.

Ameloblastoma in Jamaica - predominantly unicystic: analysis of 47 patients over a 16-year period and a case report on re-entry cryosurgery as a new modality of treatment for the prevention of recurrence / El ameloblastoma en Jamaica - predominantemente unicístico: análisis de 47 pacientes por un período de más de 16 años y un reporte de caso de criocirugía de reentrada como nueva modalidad de tratamiento para prevenir la recurrencia

OBJECTIVE: To analyse all the cases of ameloblastoma seen in the two major public Hospitals in Jamaica over a 16year period, and to compare the results with what has been previously documented by other authors in the literature. A new treatment modality for prevention of recurrence of ameloblastoma is described. SUBJECTS AND METHODS: The case files of patients histologically diagnosed to have ameloblastoma at both the Kingston Public Hospital and Cornwall Regional Hospital in Jamaica from 1980 to 1995 were retrieved and information about this odontogenic tumour was documented. The analysis revealed that uncystic ameloblastoma was predominant (95%) and solid ameloblastoma was about 5%. It was not therefore surprising that the average age in the study was 29.1 years, with a peak in the 10-19year age group. No peripheral ameloblastoma was diagnosed. RESULTS: A total of 47 new cases of ameloblastoma was recorded between 1980 and 1995. This number accounted for 16.03% of all jaw bone lesions in Jamaica and 38.2% of jaw bone lesions of odontogenic origin during this period. These cases of ameloblastoma accounted for 67% of odontogenic tumours with the exception of the odontomas. The mean age was 29.1 (range 13-67) years. The male: female ratio was 1: 1.14). Only 3 cases involved the maxilla. Of significance, is the case of bilateral ameloblastoma with no continuity to the anterior region, and also the case of unilateral involvement of the mandible and the maxilla in the same patient. Reentry cryosurgery (a procedure in which cryosurgery is done after a specified period from the primary surgical procedure) after excisional biopsy of a relatively small cystic ameloblastoma of the maxilla was negative for recurrence. CONCLUSION: The clinicopathological presentation of ameloblastoma in Jamaica is not different from what has been documented by other authors throughout the world; however, we report two unique cases - a case of bilateral ameloblastoma of the mandible and a case of unilateral involvement of the mandible and maxilla in the same patient. We suggest that reentry cryosurgery before recurrence be considered in the management of ameloblastoma for the prevention of recurrence.

OBJETIVO: Analizar todos los casos de ameloblastoma vistos en los dos principales hospitales públicos en Jamaica durante un período de 16 años, y comparar los resultados con la información previamente documentada por otros autores en la literatura. Se describe una nueva modalidad de tratamiento para prevenir la recurrencia del ameloblastoma. SUJETOS Y MÉTODO: Se recuperaron los expedientes de los pacientes diagnosticados histológicamente con ameloblastoma en el Hospital Público de Kingston y el Hospital Regional de Cornwall en Jamaica desde 1980 a 1995, y se obtuvo información documentada acerca de este tumor odontogénico. El análisis reveló que el ameloblastoma unicístico era predominante (95%), en tanto que el ameloblastoma sólido fue de alrededor del 5%. Por lo tanto, no fue sorprendente que la edad promedio en el estudio fuera 29.1 años, con un pico en el grupo etario de 10 a 19 años. No se diagnosticó ningún ameloblastoma periférico. RESULTADOS: Entre 1980 y 1995, se registró un total de 47 nuevos casos de ameloblastoma. Este número representaba el 16.03% de todas las lesiones óseas de mandíbula en Jamaica y 38,2% de las lesiones de hueso de la mandíbula de origen odontogénico durante este período. Estos casos de ameloblastoma representaron el 67% de los tumores odontogénicos con la excepción de los odontomas. La edad promedio fue 29.1 años (rango 13-67). La proporción hombre:mujer fue de 1: 1.14. Sólo tres casos implicaron el maxilar. Resulta particularmente importante el caso del ameloblastoma bilateral sin ninguna continuidad a la región anterior, así como el caso de la participación unilateral de la mandíbula y el maxilar en el mismo paciente. La criocirugía de reentrada (un procedimiento en el cual la criocirugía se realiza después de un período especificado a partir del procedimiento quirúrgico primario) después de la biopsia por escisión de un ameloblastoma quístico del maxilar relativamente pequeño, fue negativa para la recurrencia. CONCLUSIÓN: La manifestación clínicopatológica del ameloblastoma en Jamaica no difiere de lo que ha sido documentado por otros autores en todo el mundo. Sin embargo se reportan dos casos únicos: un caso de ameloblastoma bilateral de la mandíbula y un caso de compromiso unilateral de la mandíbula y el maxilar en el mismo paciente. Sugerimos que se tenga en cuenta la criocirugía de reentrada a la hora de tratar el ameloblastoma para prevenir la recurrencia.

Síndrome respiratorio agudo grave. El laboratorio de salud pública frente a una emergencia global / Severe acute respiratory syndrome: the public health laboratory in a global emergency

By the end of year 2002 there was an outbreak of atypical pneumonia in Southeast Asia which soon spread to other continents. This new severe acute respiratory syndrome (SARS) was produced by a novel coronavirus. Due to the severity of the situation and risk of introduction of this pathology in our country, the need to arrange specific laboratory diagnostic tests arose. Classic techniques, such as the electron microscopy and molecular biology test such as retrotranscription followed by the polymerase chain reaction (RT-PCR) were implemented. The araldit included cells infected with bovine coronavirus which allowed the viral particles to be visualized easily but it took more time in comparison with the negative staining of free particles from viral cultures. RT-PCR was able to detect RNA of isolated viruses from cases in Hong Kong and Germany.

A fines del año 2002 se inicia un brote de neumonía atípica en el Sudeste asiático el cual se extiende posteriormente a otros continentes. El nuevo síndrome respiratorio agudo grave (SARS) era producido por un coronavirus novedoso. Debido a la gravedad de la situación y al riesgo de introducción de esta patología en Argentina, se implementaron técnicas de diagnóstico clásicas como la microscopía electrónica, y moleculares como una reacción de retrotranscripción seguida de una reacción en cadena de la polimerasa (RT-PCR). La inclusiónen araldita de células infectadas con un coronavirus bovino permitió visualizar más fácilmente las partículas virales, pero requirió más tiempo en comparación con la coloración negativa de partículas libres de cultivos virales.La RT-PCR implementada fue capaz de detectar ARN de cepas de casos de Hong Kong y de Alemania.