Ação antimicrobiana e antibiofilme dos extratos combinados de canela e própolis sobre cepas clínicas de Acinetobacter baumannii e Pseudomonas aeruginosa multirresistentes / Antimicrobial and antibiofilm action of combined extracts of cinnamon and propolis on multiresistant clinical strains of Acinetobacter baumannii and Pseudomonas aeruginosa

Os microrganismos resistentes a diferentes classes de agentes antimicrobianos têm se tornado cada vez mais comuns e atualmente são denominados como multirresistentes. Nos hospitais, tais microrganismos apresentam maior perigo, pois são causadores de infecções nosocomiais e a higienização bucal deficiente dos pacientes internados pode tornar a cavidade bucal um sítio para proliferação desses microrganismos multirresistentes. Diante do exposto, novos compostos com ação antimicrobiana precisam ser estudados. O objetivo deste estudo foi avaliar quimicamente o extrato hidroalcóolico de própolis verde de Baccharis dracunculifolia e de Cinnamomum verum (canela) que foram obtidos a partir da extração da matériaprima, analisar a atividade antimicrobiana e antibiofilme dos extratos isolados e combinados contra quatro cepas clínicas multirresistentes de Pseudomonas aeruginosa e Acinetobacter baumannii e verificar a citotoxicidade dos produtos vegetais in vitro em linhagem celular de queratinócitos humanos (HaCat). Para tanto, os extratos vegetais foram preparados a partir da matéria-prima da canela em casca e da própolis bruta. Em seguida, foram caracterizados quimicamente por cromatografia líquida de alta eficiência (HPLC-DAD) para identificação dos principais compostos e a análise do teor de sólidos solúveis dos extratos vegetais também foi realizada. Para avaliação antimicrobiana, foram performados o teste de microdiluição em caldo de acordo com a Clinical and Laboratory Standards Institute (CLSI) e a análise de Checkerboard, para avaliar o efeito combinado dos extratos. A atividade antibiofilme dos extratos combinados foi realizada por meio do teste de MTT, no qual diferentes tempos de contato (5 e 30 min) e diferentes modalidades (inibição na formação do biofilme bacteriano e erradicação do biofilme bacteriano já formado) foram testadas. Para ação citotóxica, as células foram cultivadas em meio DMEM e semeadas na placa de 96 poços. Após aderência inicial, aplicou-se os extratos em diferentes concentrações baseadas nas análises microbiológicas para avaliação da viabilidade celular por meio do teste de MTT. Os dados foram analisados por ANOVA e teste de Tukey, ou Kruskal-Wallis e Dunn, considerando um nível de significância de 5%. Os compostos identificados no extrato de própolis verde de B. dracunculifolia foram ácido clorogênico, derivado do ácido cinâmico e apigenina. O aldeído cinâmico foi o principal composto identificado no extrato de C. verum. Os extratos vegetais apresentaram ação bactericida sobre todas as cepas analisadas e, quando combinados, os extratos atuaram de modo aditivo e algumas combinações sinérgicas foram encontradas. O protocolo de inibição da formação do biofilme promoveu percentuais de redução superiores quando comparado ao protocolo de erradicação. Valores expressivos de 83,86% (p < 0,05) de inibição da formação de biofilme de uma cepa clínica de A. baumannii e 89,31% (p < 0,05) de inibição em uma cepa clínica de P. aeruginosa foram encontrados com a aplicação dos extratos combinados. A atuação dos produtos vegetais foi estatisticamente semelhante a atuação da clorexidina 0,12%. Em conclusão, os extratos de própolis verde e canela na forma isolada ou combinada apresentaram ação antimicrobiana e antibiofilme sobre cepas clínicas de A. baumannii e P. aeruginosa multirresistentes. Dessa forma, os produtos vegetais são promissores agentes antissépticos para futuras formulações odontológicas. (AU)

Microorganisms resistant to different classes of antimicrobial agents have become increasingly common and are currently called multidrug resistant. In hospitals, such microorganisms are more dangerous, as they cause nosocomial infections and poor oral hygiene in hospitalized patients can make the oral cavity a site for the proliferation of these multiresistant microorganisms. Given the above, new compounds with antimicrobial action need to be studied. The objective of this study was to chemically evaluate the hydroalcoholic extract of green propolis from Baccharis dracunculifolia and Cinnamomum verum (cinnamon) that were obtained from the extraction of the raw material, to analyze the antimicrobial and antibiofilm activity of the isolated and combined extracts against four clinical strains multiresistant strains of Pseudomonas aeruginosa and Acinetobacter baumannii and verify the cytotoxicity of plant products in vitro in human keratinocyte cell lineage (HaCat). For this purpose, plant extracts were prepared from raw cinnamon bark and raw propolis. Then, they were chemically characterized by high performance liquid chromatography (HPLC-DAD) to identify the main compounds and the analysis of the soluble solids content of the plant extracts was also performed. For antimicrobial evaluation, the broth microdilution test according to the Clinical and Laboratory Standards Institute (CLSI) and the Checkerboard analysis were performed to evaluate the combined effect of the extracts. The antibiofilm activity of the combined extracts was performed using the MTT test, in which different contact times (5 and 30 min) and different modalities (inhibition of bacterial biofilm formation and eradication of already formed bacterial biofilms) were tested. For cytotoxic action, cells were cultured in DMEM medium and seeded in the 96-well plate. After initial adhesion, the extracts were applied at different concentrations based on microbiological analyzes to assess cell viability through the MTT test. Data were analyzed by ANOVA and Tukey's test, or Kruskal-Wallis and Dunn, considering a significance level of 5%. The compounds identified in the green propolis extract of B. dracunculifolia were chlorogenic acid, cinnamic acid derivative and apigenin. Cinnamic aldehyde was the main compound identified in the C. verum extract. The plant extracts showed bactericidal action on all strains analyzed and, when combined, the extracts acted additively and some synergistic combinations were found. The biofilm formation inhibition protocol promoted higher reduction percentages when compared to the eradication protocol. Significant values of 83.86% (p < 0.05) inhibition of biofilm formation in a clinical strain of A. baumannii and 89.31% (p < 0.05) inhibition in a clinical strain of P. aeruginosa were found with the application of the combined extracts. The performance of plant products was statistically similar to the performance of 0.12% chlorhexidine. In conclusion, extracts of green propolis and cinnamon, in isolated or combined form, showed antimicrobial and antibiofilm action on multiresistant clinical strains of A. baumannii and P. aeruginosa. Thus, plant products are promising antiseptic agents for future dental formulations. (AU)

Chemical constituents from the leaves of Cinnamomum camphora var. linaloolifera and their anti-inflammatory activities / 中国中药杂志

Thirteen compounds were isolated and purified from the leaves of Cinnamomum camphora by the macroporous resin,silica gel,and Sephadex LH-20 column chromatographies. Those compounds were further identified by IR,UV,MS,and NMR techniques:( 2 S)-1-( 3″,4″-methylenedioxy phenyl)-3-( 2',6'-dimethoxy-4'-hydroxyphenyl)-propan-2-ol( 1),( 2 R,3 R)-5,7-dimethoxy-3',4'-methylenedioxy flavanol( 2),9-hydroxysesamin( 3),sesamin( 4),piperitol( 5),kobusin( 6),(-)-aptosimon( 7),acuminatolide( 8),1β,11-dihydroxy-5-eudesmene( 9),lasiodiplodin( 10),vanillin( 11),p-hydroxybenzaldehyde( 12),and p-hydroxybenzoic acid ethyl ester( 13). Compound 1 was a novel compound,and compounds 2,6,7,9 and 10 were isolated from Cinnamomum plants for the first time. Compounds 4,7 and 10 were found to possess good inhibitory effect on IL-6 production in LPS-induced BV2 cells at a concentration of 20 μmol·L-1 in the in vitro bioassay,with inhibition rates of 51. 26% ± 4. 13%,67. 82% ± 3. 77% and85. 81%±1. 19%,respectively.

Avaliação in vitro da atividade de óleos essenciais sobre Candida auris / In vitro evaluation of the activity of essential oils about Candida auris

O objetivo deste trabalho foi avaliar a atividade in vitro dos óleos essenciais de Cinnamomum cassia, Eucalyptus citriodora; Eucalyptus globulus, Eugenia caryophyllus, Melaleuca alternifólia, Myristica fragans, Ocimum basilicum, Origanum majorana, Origanum vulgare, Rosamarinus officinalis, Salvia sclarea, Schinus terebinthifolius, Thymus vulgaris e Zingiber officinale, bem como dos compostos químicos desses óleos essenciais, com as menores concentrações fungicidas mínimas sobre três isolados biológicos de Candida auris de origem humana. Determinar e avaliar in vitro as concentrações fungicidas mínimas dos antifúngicos sintéticos anfotericina B, cetoconazol, flucitocina, fluconazol, itraconazol e voriconazol sobre Candida auris. Para avaliação da atividade antifúngica e obtenção da concentração fungicida mínima (CFM) utilizou-se a técnica de microdiluição em caldo, na base 2, em meio RPMI 1640, acrescido de tensoativo Tween20, e a confirmação da inibição em Agar Sabouraud dextrose, a 37ºC por 24hs. Para a avaliação da sensibilidade a anfotericina B, cetoconazol, flucitocina...(AU)

The objective of this search was to evaluate the in vitro activity of the essential oils Cinnamomum cassia, Eucalyptus citriodora; Eucalyptus globulus, Eugenia caryophyllus, Melaleuca alternifolia, Myristica fragans, Ocimum basilicum, Origanum majorana, Origanum vulgare, Rosamarinus officinalis, Salvia sclarea, Schinus terebinthifolius, Thymus vulgaris and Zingiber officinale as well as the chemical compounds of these essencial oils with the lowest minimum fungicidal concentrations on three biological isolates of Candida auris of human origin. Determine and evaluate in vitro the minimum fungicidal concentrations of the synthetic antifungals amphotericin B, ketoconazole, flucitocin, fluconazole, itraconazole and voriconazole on Candida auris. To evaluate the antifungal activity and obtain the minimum fungicidal concentration (CFM), the broth microdilution technique was used in RPMI 1640 medium, plus Tween20 surfactant, and confirmation of inhibition in Sabouraud dextrose Agar, at 37ºC for 24 hours. For the evaluation of sensitivity to amphotericin B, ketoconazole, flucitocin, fluconazole, itraconazole and voriconazole, the commercial test E-test® was used. The three isolates were resistant to the oils Myristica fragans, Shinus terebinthifolius and Zenziber officinale. The other essential oils showed inhibitory activity on Candida auris. The three isolates were more sensitive to Cinnamomum cassia essential oil, with CFM of 0.00001 µg/mL for isolate 1 and CFM of 0.10 µg/mL for isolate 2 and CFM of 0.05 µg/mL for isolate 3. Thymus vulgaris also showed lower fungicidal concentration for isolates 1 and 2, with CFM results of 684.76 µg/mL for isolate 1 and CFM of 171.19 µg/mL for isolate 2. The three biological isolates evaluated were more sensitive to cinnamic aldehyde, isoeugenol and timol, with the following results being found for isolates 1, 2 and 3 respectively: Cinnamic aldehyde - CFM 0.026µg/mL, 0.83 µg/mL and 3, 33 µg/ml; Isoeugenol CFM of 1.60 µg/ml, 1.60 µg/ml and 12.82 µg/ml; timol CFM of 2.93 µg/ml, 1.46 µg/ml and 11.74 µg/ml. The three isolates were resistant to chemical compounds: α-Humulene, ßCaryophyllene, γ-Terpinene and P-Cimene. For synthetic antifungal agents, 7 the three isolates were resistant to fluconazole and voriconazole. The MIC of amphotericin B was 0.50 µg / ml for isolates 1 and 3 and 0.75 µg / ml for isolate 2; for ketoconazole, MIC was 3 µg / ml for isolates 1 and 2 and 4 µg / ml for isolate 3; the MIC of flucitocin was 2 µg / ml for isolate 1, 1 µg / ml for isolate 2 and 1.5 µg / ml for isolate 3; for itraconazole the MIC was 4 µg / ml for the three isolates. According to the conditions of this research, the essential oils of Cinnamomum cassia, Thymus vulgaris and Origanum vulgare and the chemical compounds cinnamic aldehyde, isoeugenol and thymol showed antifungal activity with the lowest concentrations of fungicides in biological isolates of Candida auris. (AU)

Advances in quality research of Cinnamomum cassia / 中国中药杂志

Cinnamomum cassis is one of the commonly used traditional Chinese medicines in China. Its genuine producing areas distribute in Guangdong and Guangxi provinces. As an important edible herb and export variety of China, the quality control and internationalization of quality standards of C. cassis is extremely significant. In the recent years, with the development of the cinnamon industry, relevant academic research and the upgrade of the international standards, it is necessary to summarize the quality-related progress of C. cassis. In the present review, the germplasm resources, specific quality marker(Q-marker) and quality standards of C. cassis were summarized on the basis of published research during the last 10 years.

Adaptação de Staphylococcus aureus ao óleo essencial de Cinnamomum cassia / Adaptation of Staphylococcus aureus to Cinnamomum cassia essential oil

O objetivo deste trabalho foi avaliar a capacidade de adaptação de Staphylococcus aureus ao óleo essencial (OE) de canela cássia (Cinnamomum cassia). Primeiro determinou-se a Concentração Mínima Bactericida (CMB) do OE de C. cassia utilizando a técnica de microdiluição. Em seguida as células de S.aureus foram expostas a concentrações subletais do OE de C. cassia(CMB/4 e CMB/8) por um período de 6h e testadas frente a diferentes concentrações do OE (CMB/2; CMB; 1,2CMB; 1,4CMB; 1,6CMB; 1,8CMB e 2CMB), estas foram incubadas e plaqueadas em TSA (Ágar Triptona de Soja) empregando a técnica de microgotas. As células de S.aureus apresentaram uma nova CMB, maior que a anterior, portanto foram classificadas como capazes de se adaptarem ao OE de C. cassia. Após a exposição a concentrações subletais do OE, a CMB foi de 0,0744%. Os resultados evidenciam a importância de se utilizar a concentração adequada do antimicrobiano para evitar a adaptação do microrganismo.

Improvement in histology, enzymatic activity, and redox state of the liver following administration of Cinnamomum zeylanicum bark oil in rats with established hepatotoxicity / 대한해부학회지

Formaldehyde (FA) is an environmentally-available pollutant. Since the liver acts as a detoxifier in the human body, it is the first and most affected organ in individuals exposed to higher-than-normal amounts of FA. FA mainly alters oxidant/antioxidant status and initiates oxidative stress, and by means, causes functional damage to the liver. Thus, it is important to identify natural bioactive compounds with antioxidant properties in order to be used as food additives. Cinnamon (Cinnamomum zeylanicum) is a popular flavor and also a medicinal plant with a variety of beneficial effects. In the present original study, cinnamon essential oil (CEO) has been administrated at doses of 10, 20, and 100 mg/kg, orally, to hepatotoxicity rat models caused by FA (10 mg/kg, intraperitoneally). Liver enzymes and its histology were assessed and oxidative stress biomarkers in the liver tissue were also examined. CEO administration caused a significant increase in superoxide dismutase, glutathione peroxidase, and catalase and a prominent decrease in nitric oxide levels in the liver tissue. Also, in serum samples, CEO significantly reduced the elevated amounts of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase. When assessed histologically, portal area and central vein fibrosis alongside with the hepatocytes' hypereosinophilia and swelling, focal inflammation, and necrotic areas were found to be prominently decreased in the CEO group. In conclusion, our study suggested that the CEO may have the potential for being used against FA-induced hepatotoxicity.

Molecular mechanism of Cinnamomi Ramulus-Paeoniae Radix Alba drug pair against sepsis based on integrative pharmacology platform of traditional Chinese medicine / 中国中药杂志

Guizhi Decoction is a resolving agent,which is a classic prescription for traditional Chinese medicine. It is effective in the treatment of sepsis in clinical practice. However,due to the complexity of the prescription,its anti-sepsis mechanism is difficult to be clarified. The " Cinnamomi Ramulus-Paeoniae Radix Alba" drug pair,as the classic compatibility for medicinal and medicinal herbs,is the core of Guizhi Decoction. In this study,Cinnamomi Ramulus-Paeoniae Radix Alba drug pair was used as the research object and the molecular mechanism of its treatment of sepsis was investigated by analyzing the chemical compositions with integrative pharmacology platform( TCMIP,http://www.tcmip.cn/),predicting disease target,analyzing gene function and pathway of " Cinnamomi Ramulus-Paeoniae Radix Alba" in treatment of sepsis,and establishing a multi-dimensional network relationship of " Chinese medicine-chemical components-core targets-key pathways". The prediction results of " Cinnamomi Ramulus-Paeoniae Radix Alba" drug pair showed that its anti-sepsis effect was associated with 45 active components,and the active components played an anti-sepsis role through multiple targets and pathways,involving inflammatory targets such as PF4,MyD88,TLR4,BDKRB2,CD14,and NOS3. The sepsis was relieved mainly by regulating Toll like signaling pathway,Fox O signaling pathway,chemokines signaling pathway,thyroid and insulin endocrine signaling pathways and biological processes. This study provides a scientific basis for further development of Cinnamomi Ramulus-Paeoniae Radix Alba drug pair and Guizhi Decoction against sepsis.

Mechanism of Cinnamomi Ramulus improving rat model of intrahepatic cholestasis induced by ANIT by regulating FXR pathway / 中国中药杂志

To study the mechanism and action of Cinnamomi Ramulus in ameliorating intrahepatic cholestasis induced by α-isothiocyanate( ANIT) in rats by regulating FXR pathway. Forty SD rats were randomly divided into normal group,model group,positive control( ursodeoxycholic acid) group( 60 mg·kg~(-1)),Cinnamomi Ramulus treatment( 60 mg·kg~(-1)·d~(-1)) group,and Cinnamomi Ramulus treatment( 20 mg·kg~(-1)·d~(-1)) group,with 8 rats in each group. Except for the normal control group,the other groups were intragastrically administered with the corresponding concentrations of continuous aqueous solution( 0. 005 m L·g~(-1)),once a day,for 7 days.Except for the normal group,the other groups were treated with ANIT( 100 mg·kg~(-1)),once a day,for 3 days. Blood was taken from the abdominal aorta 24 hours after the last administration,and serum alanine aminotransferase( ALT),aspartate aminotransferase( AST),total bilirubin( TBi L),and total bile acid( TBA) were measured. 1. 5-2 cm of rat liver tissue was taken. After fixation with10% formaldehyde,paraffin-embedded sections were taken,HE staining was performed,and immunohistochemistry( IHC) was used to analyze the expression of FXR. RNA and protein were extracted from rat liver tissue to detect FXR mRNA expression,as well as bile acid synthesis and detoxification,transport related SHP,UGT2 B4,BSEP protein expressions at downstream of FXR. Compared with the normal group,serum ALT,AST,TBi L,and TBA levels were elevated in the model group( P<0. 01),liver damage was severe,FXR protein's optical density decreased,FXR mRNA expression decreased,and SHP,UGT2 B4,BSEP protein expressions were decreased( P<0. 05,P<0. 01). Compared with the model group,the drug group could reduce serum ALT,AST,TB,TBA levels to different degrees( P<0. 05,P<0. 01),alleviate liver tissue damage,increase the optical density of FXR protein,and promote the expressions of FXR mRNA and FXR,SHP,BSEP and UGT2 B4 proteins( P<0. 05,P<0. 01). Cinnamomi Ramulus can alleviate ANIT-induced intrahepatic cholestasis,and reduce hepatocyte injury and serum ALT,AST,TBi L and TBA levels. The mechanism may be through FXR-SHP,FXR-UGT2 B4,FXR-BSEP signaling pathways. Therefore,in the pathogenesis of intrahepatic cholestasis,we can try to further explore in alleviating intrahepatic cholestasis with Cinnamomi Ramulus,so as to provide effective drugs for clinical treatment of intrahepatic cholestasis.

Transcriptome analysis of Cinnamomum longepaniculatum by high-throughput sequencing

Background: Cinnamomum longepaniculatum is an important commercial crop and the main source of volatile terpenoids. The biosynthesis of key bioactive metabolites of C. longepaniculatum is not well understood because of the lack of available genomic and transcriptomic information. To address this issue, we performed transcriptome sequencing of C. longepaniculatum leaves to identify factors involved in terpenoid metabolite biosynthesis. Results: Transcriptome sequencing of C. longepaniculatum leaves generated over 56 million raw reads. The transcriptome was assembled using the Trinity software and yielded 82,061 unigenes with an average length of 879.43 bp and N50 value of 1387 bp. Furthermore, Benchmarking Universal Single-Copy Orthologs analysis indicated that our assembly is 91% complete. The unigenes were used to query the nonredundant database depending on sequence similarity; 42,809 unigenes were homologous to known genes in different species, with an annotation rate of 42.87%. The transcript abundance and Gene Ontology analyses revealed that numerous unigenes were associated with metabolism, while others were annotated in functional categories including transcription, signal transduction, and secondary metabolism. The Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that 19,260 unigenes were involved in 385 metabolic pathways, with 233 unigenes found to be involved in terpenoid metabolism. Moreover, 23,463 simple sequence repeats were identified using the microsatellite identification tool. Conclusion: This is the first detailed transcriptome analysis of C. longepaniculatum. The findings provide insights into the molecular basis of terpenoid biosynthesis and a reference for future studies on the genetics and breeding of C. longepaniculatum.

Effect of a mixture of Galla rhois and Cinnamomum cassia extracts on susceptibility to the colonization of Campylobacter jejuni in broiler chickens

The present study evaluated the effects of a mixture of Galla rhois and Cinnamomum cassia extracts (GCE) (1 : 1, w/w) on susceptibility to the colonization of Campylobacter (C.) jejuni in broilers. Eighty two-week-old broilers (n = 20 per group) were used to estimate the efficacy of GCE against C. jejuni infection via drinking water. Antibacterial activity testing revealed that the minimum bactericidal concentration of GCE against C. jejuni was 2.5 mg/mL. Broilers challenged with C. jejuni were administered 0.0 (Non-GCE), 2.5 (GCE-2.5), 5.0 (GCE-5.0) and 10.0 g/L (GCE-10) GCE for 7 days, and the cecal contents were collected from five broilers per group on the 1st, 3rd, 5th, and 7th day post-treatment. On day 3 post-administration, the number of C. jejuni in GCE-5.0 (p < 0.05) and GCE-10 (p < 0.01) was significantly decreased relative to Non-GCE, while on day 7 those in all GCE-treated groups were significantly decreased compared to the Non-GCE group (p < 0.001). Hematological and blood biochemical analysis revealed no significant differences in parameters between the Non-GCE and GCE-treated groups. Based on the results of the present study, GCE was identified as a safe and alternative candidate to suppress C. jejuni colonization in broilers.

Antibacterial activity of α-terpineol may induce morphostructural alterations in Escherichia coli

The antibacterial effect of α-terpineol from Cinnamomum longepaniculatum (Gamble) N. Chao leaf essential oils were studied with special reference to the mechanism of inhibiting the standard strain of Escherichia coli (CMCC (B) 44102) growth at ultrastructural level. Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) and time-kill curves of α-terpineol were determined; Escherichia coli was treated with α-terpineol and observed under a transmission electron microscope. The MIC and MBC values of α-terpineol were all 0.78 µL/mL, and time-kill curves showed the concentration-dependent. Under the transmission electron microscopy (TEM), Escherichia coli exposed to MIC levels of α-terpineol exhibited decreased cell size and irregular cell shape, cell wall and cell membrane were ruptured, nucleus cytoplasm was reduced and nuclear area gathered aside. Results suggest that α-terpineol has excellent antibacterial activity and could induce morphological changes of Escherichia coli.

In vitro culture of immature embryos of Cinnamomum tamala Nees.— The role of different factors.

Seed characteristics and in vitro culture of C. tamala embryos were studied. Embryos desiccated below 50% (fresh weight) exhibited poor morphogenetic response in vitro and confirmed the recalcitrant nature of seeds. The immature embryos of various developmental ages (4-16 week after flowering, WAF) were cultured on different strengths of MS medium. Morphogenesis responses were recorded after 10 days of culture. The best culture responses were achieved from the immature embryos of 12 WAF on MS medium with sucrose (3%, w/v), polyvinyl pyrollidone (100 mg L-1) and benzyl adenine (12 µM). Under optimum condition ~60% explants responded; and ~7.3 shoots buds developed per explants after 35 days of culture initiation. The shoot buds could be converted into micro-shoots on MS medium with sucrose (3%) and kinetin (3 µM). About 5.3 micro-shoots/shoot buds sprouted per sub-culture. The micro-shoots were rooted by maintaining them on MS medium with α-naphthalene acetic acid (3 µM) where within 6-8 wk of culture ~8-10 roots developed. The rooted plantlets were acclimatized in vitro before they were transferred to community potting mix and maintained in the poly-shade ca 75% shading. The transplants registered ~70% survival after two months of transfer.

Study on multi-target optimization of prescription dose of Mahuang decoction / 中国中药杂志

<p><b>OBJECTIVE</b>To optimize the prescription dose of Mahuang decoction in a multi-target manner, in order to provide reference for the quantitative optimization of the prescription dose of the traditional Chinese medicine compound.</p><p><b>METHOD</b>The number of diaphoretic spots in rats, the tracheal antispasmodic rate in guinea pigs and the writhing times by acetic acid in mice were taken as the indexes for evaluating the diaphoretic, antispasmodic and analgesic effects. According to the experimental results of the 16 orthogonal combination prescriptions, a mathematical dose-effect model was built by support vector regression (SVR) and quadratic response surface regression (RSR) respectively. The multi-target optimization was achieved by elitist non-dominated sorting genetic algorithm (NSGA-II) and entropy weight TOPSIS method.</p><p><b>RESULT</b>The optimal dose of Mahuang decoction after being optimized by SVR modeling contained 17.71 g of Ephedrae Herba, 9.57 g of Cinnamomi Ramulus, 11.75 g of Armeniacae Semen Amarum and 4.39 g of Glycyrrhizae Radix et Rhizoma Praeparata Cum Melle. The optimized result by RSR modeling contained 13.37 g of Ephedrae Herba, 11.61 g of Cinnamomi Ramulus, 11.98 g of Armeniacae Semen Amarum and 5.67 g of Glycyrrhizae Radix et Rhizoma Praeparate Cum Melle. SVR was superior to RSR in both of the forecast capacity and optimization results.</p><p><b>CONCLUSION</b>SVR-NSGA-II-TOPSIS method could be adopted for the multi-target optimization for the dose of Mahuang decoction and other traditional Chinese medicine compounds. It is proved to be the optimal prescription with the best efficacy, and could provide scientific quantitative basis for determining the dose of traditional Chinese medicine compound prescriptions and developing new traditional Chinese medicines.</p>

Study on preparation process and stability of beta-cyclodextrin inclusion compound in volatile oil of Cinnamomum longepaniculatum leaves / 中国中药杂志

To study the optimum preparation process and stability of beta-cyclodextrin inclusion compound in volatile oil of Cinnamomum longepaniculatum leaves. The saturated aqueous solution method was adopted to prepare inclusion compounds for an orthogonal test. The inclusion compound productivity and the inclusion rate were taken as indexes for screening the inclusion processes. The inclusion effect was evaluated with the infrared spectrophotometry and TLC, and the stability under conditions of high temperature, high humidity and strong light was detected. Under optimum preparation conditions for inclusion, the ratio between volatile oil and beta-cyclodextrin was 1: 8 (mL: g), that between beta-cyclodextrin and water was 1: 15, the inclusion temperature was 40 degrees C, and the inclusion time was 3 h. The results of spectrophotometry and TLC showed that the optimum conditions can generate beta-cyclodextrin inclusion compound in volatile oil of C. longepaniculatum leaves with certain light resistance, thermo-stability and hygro-stability. Therefore the optimum inclusion process features simple operation and stable inclusion compounds.

Preparation of cinnamomi cortex oil microspheres based on porous silicon dioxide and its property characterizations / 中国中药杂志

To determine the optimum process for preparing Cinnamomi Cortex oil microspheres based on porous silicon dioxide. After porous silica dioxide adsorbed Cinnamomi Cortex oil, Cinnamomi Cortex oil microspheres were prepared by the dropping method, with sodium alginate as the skeleton materials. The preparation process was optimized through the L(9) (3(4)) orthogonal test design, with microspheres diameter, distribution, drug loading capacity and entrapment efficiency as the indexes. The cinnamon volatile oil microspheres were characterized by scanning election microscope (SEM), thermogravimetric analysis (TGA), and infrared (IR) spectroscopy. An in vitro drug release experiment was conducted. The results showed that the microspheres prepared with the optimal process parameters were in good shape, even in size and good in dispersibility, with an average diameter of 1.61 mm, an average drug loading capacity of 32.85%, an entrapment efficiency of 94.79%. The maximum drug release capacity reached 72.6%, 95.0%, 97.4%, respectively, under pH 4.0, 6.8, 7.4 in 6 hours. Meanwhile, microsphere generation was tested by IR, TGA and other methods. The established optimum process for preparing Cinnamomi Cortex oil microspheres was proved to be stable and practical.

Chemical constituents of Cinnamomum cebuense / 中国天然药物

AIM@#To investigate the chemical constituents of Cinnamomum cebuense, an endemic and critically endangered tree found only in Cebu, Philippines.@*METHODS@#The compounds were isolated by silica gel chromatography. The structures of the isolates were elucidated by NMR spectroscopy.@*RESULTS@#The dichloromethane (DCM) extract of the bark of C. cebuense afforded a new monoterpene natural product 1 and a new sesquiterpene 2, along with the known compounds, 4-hydroxy-3-methoxycinnamaldehyde (3), 4-allyl-2-methoxyphenol (4), α-terpineol (5) and humulene (6). The DCM extract of the leaves of C. cebuense yielded 6, β-caryophyllene (7), squalene (8), and a mixture of α-amyrin (9), β-amyrin (10) and bauerenol (11). The structures of 1-7 were elucidated by extensive 1D and 2D NMR spectroscopy, while the structures of 8-11 were identified by comparison of their (13)C NMR data with those reported in the literature.@*CONCLUSION@#The bark of C. cebuense afforded monoterpenes, sesquiterpenes and phenolics, while the leaves yielded sesquiterpenes and triterpenes.

Study on fingerprints of chemical constituents of cinnamomi ramulus and cinnamomi cortex / 中国中药杂志

<p><b>OBJECTIVE</b>To compare the difference of chemical constituents of Cinnamomi Ramulus and Cinnamomi Cortex,and study the characteristics of their chemical constituents, in order to provide basis for clinical application and quality control.</p><p><b>METHOD</b>HPLC fingerprint method was used to analyze chemical constituents of Cinnamomi Ramulus and Cinnamomi Cortex compare the difference between.</p><p><b>RESULT</b>Cinnamomi Ramulus and Cinnamomi Cortex had similar HPLC fingerprint profiles, but with difference in peak areas of their chemical constituents. Specifically, the average peak areas of cinnamyl alcohol, cinnamic acid and 2-methoxy cinnamic acid were higher than that of Cinnamomi Cortex, whereas the average peak areas of 2-hydroxyl cinnamaldehyde, coumarin, cinnamaldehyde and 2-methoxy cinnamaldehyde in Cinnamomi Cortex were higher than that of Cinnamomi Ramulus.</p><p><b>CONCLUSION</b>This method can be used for evaluating quality of Cinnamomi Ramulus and Cinnamomi Cortex. The ratio between the peak areas of cinnamaldehyd and cinnamic acid can be used to discriminate most Cinnamomi Ramulus samples (<23) and Cinnamomi Cortex samples (>23). Cinnamomi Ramulus and Cinnamomi Cortex have the similar types of chemical constituents but with difference in content. This study provides reference for the pharmacodynamic difference of Cinnamomi Ramulus and Cinnamomi Cortex.</p>

In vitro antibacterial effects of five volatile oil extracts against intramacrophage Brucella abortus 544

Brucella abortus is a gram-negative facultative intracellular bacterium that can cause a highly contagious disease in sheep, goats, cattle and one-humped camels. It is responsible for one of the most important zoonosis in human. The aim of this study was to evaluate the role of Mentha piperita, Origanum majorana, Citrus lemon, Cinnamomum verum and Myristica fragrans essential volatile oil extracts on human macrophages infected by B. abortus 544. Essential volatile oil extracts from M. piperita, O. majorana, C. lemon, C. verum and M. fragrans were extracted. Human macrophages were cultured at a density of 2_10[5] cells per well in sterile 96-well microtiter plates, and infected with B. abortus 544 at a ratio of 1:100 bacteria/cell. Then essential volatile oil extracts were added at a concentration of 1%. At specified times; cells were washed, lysed with 0.1% Triton, and plated on 2YT agar to determine the number of intracellular bacteria. Cinnamomum verum volatile oil at a concentration of 1% had the highest antibacterial activity against B. abortus 544 inside human macrophages. Its inhibitory effect observed from 24 h and continued till 144 h after the infection. Moreover, C. verum [0.1%] in combination with 1% concentration of M. piperita, O. majorana, C. lemon or M. fragrans volatile oil extracts produced a synergistic inhibitory effect against B. abortus 544. The results indicate that, among the five selected oil extracts, C. verum volatile oil applied either separately or in combination with other oil extracts had the most effective antimicrobial activity against Brucella

Researches on relationship between genetic differentiation and chemical variation of Cinnamomum migao / 中国中药杂志

<p><b>OBJECTIVE</b>To study the relationship between the genetic diversity and chemical variation of Cinnamomum migao.</p><p><b>METHOD</b>ISSR marker technique was used to research the genetic structure of 9 population, GC-MS was used to analyze the main ingredients of the volatile oil in C. migao.</p><p><b>RESULT</b>The analysis on the main ingredients of the volatile oil showed that there were significant or extremely significant differences in 9 populations. The minimum variation index of population was Yunnan Funing and the maximum variation index of population was Guangxi Yueye. ISSR marker analysis showed that the average of polymorphic loci percentage (P) was 42.41%, expected heterozygosity (H) was 0.181 0, Shannon's information index (I) was 0.293 8, the Nei's genetic diversity (H(s)) in the group was 0.188 9, genetic differentiation index (G(st)) was 2.269 1. The relationship between the genetic diversity and chemical variation showed that there was no significant correlation between the main ingredients of the volatile oil and 4 indexes of genetic structure of C. migao.</p><p><b>CONCLUSION</b>The genetic diversity of C. migao was relatively high at the population levels, while it is low within the population levels, the relationship between chemical variation and genetic diversity was not obvious, that may indicate that other factors causes the chemical variation of C. migao.</p>

UPLC simultaneous determination of five active components in Cinnamomi Ramulus / 中国中药杂志

A rapid UPLC method for simultaneous determination of protocatechuic acid, coumarin, cinnamic alcohol, cinnamic acid and cinnamaldehyde in Cinnamomi Ramulus was established. The optimal conditions of separation and detection were achieved on an HSS T3 Column (2.1 mm x 100 mm, 1.8 microm) eluted with a gradient of acetonitrile and 0.05% aqueous phosphoric acid, at a flow rate of 0.5 mL x min(-1), detected at 254 nm. The linear response ranges of protocatechuic acid, coumarin, cinnamic alcohol, cinnamic acid and cinnamaldehyde were 0.359-3.59 mg x L(-1) (r = 0. 999 3), 2. 834-28.34 mg x L(-1) (r = 0.999 8), 0.574-5.74 mg x L(-1) (r = 0.999 8), 2.400-24.00 mg x L(-1) (r = 0.999 9), 32.57-325.7 mg x L(-1) (r = 0.999 8), respectively (n = 6). The mean recoveries (n = 9) of the five components were 96.7%-101.0%, RSD < 2.3%. The assay demonstrates that the method has adequate accuracy and selectivity to measure the concentration of protocatechuic acid, coumarin, cinnamic alcohol, cinnamic acid and cinnamaldehyde in Cinnamomi Ramulus.