Comparison of catalytic functions and expression patterns of two pinene synthases from Wurfbainia villosa / 中国中药杂志

Wurfbainia villosa fruit is rich in volatile terpenoids, among which pinene is one of the main components and has anti-inflammatory, antibacterial, anti-tumor, and other pharmacological activities. This research group found that W. villosa fruits were rich in α-pinene by GC-MS, and terpene synthase(WvTPS63, formerly known as AvTPS1) with β-pinene as the main product was cloned and identified, but α-pinene synthase had not been identified. In this study, based on the genome data of W. villosa, we screened and found WvTPS66 with highly similar sequences to WvTPS63, identified enzyme functions of WvTPS66 in vitro, and performed a comparative analysis of sequence, catalytic function, expression pattern, and promoter with WvTPS63. Multiple sequence alignment showed that the amino acid sequences of WvTPS63 and WvTPS66 were highly similar and the conservative motif of terpene synthase was almost identical. In vitro enzymatic experiments on catalytic functions showed that both could produce pinene, and the main product of WvTPS63 was β-pinene, while that of WvTPS66 was α-pinene. Expression pattern analysis showed that WvTS63 was highly expressed in flowers, WvTPS66 was expressed in the whole plant, and the highest expression level was found in the pericarp, which indicated that it might be mainly responsible for the synthesis of α-pinene in fruits. In addition, promoter analysis revealed the presence of multiple regulatory elements related to stress response in the promoter regions of both genes. The findings of this study can provide a reference for the functional study of terpene synthase genes and new genetic elements for pinene biosynthesis.

Chemical compositions, larvicidal and antimicrobial activities of Zingiber castaneum (Škorničk. & Q.B. Nguyễn) and Zingiber nitens (M.F. Newman) essential oils

Abstract In this paper, the chemical constituents, larvicidal and antimicrobial activities of hydrodistilled essential oils from Zingiber castaneum Škorničk. & Q.B. Nguyễn and Zingiber nitens M.F. Newman were reported. The main constituents of Z. castaneum leaf were bicyclogermacrene (24.8%), germacrene D (12.9%), cis-β-elemene (11.2%) and β-pinene (10.3%), while sabinene (22.9%) and camphene (21.2%) were the significant compounds in the rhizome. However, the dominant compounds in the leaf of Z. nitens includes β-pinene (45.8%) and α-pinene (10.7%). Terpinen-4-ol (77.9%) was the most abundant compound of the rhizome. Z. castaneum rhizome oil displayed larvicidal activity against Aedes aegypti and Culex quinquefasciatus with LC50 values of 121.43 and 88.86 µg/mL, respectively, at 24 h. The leaf oil exhibited activity with LC50 values of 39.30 µg/mL and 84.97 µg/mL, respectively. Also, the leaf and rhizome oils of Z. nitens displayed greater larvicidal action towards Ae. aegypti with LC50 values of 17.58 µg/mL and 29.60 µg/mL, respectively. Only the rhizome oil displayed toxicity against Cx. quinquefasciatus with LC50 value of 64.18 µg/mL. All the studied essential oils inhibited the growth of Pseudomonas aeruginosa ATCC25923 with minimum inhibitory concentration (MIC) value of 50.0 µg/mL. This paper provides information on the larvicidal and antimicrobial potentials of Z. castaneum and Z. nitens essential oils.

Chemical Composition and Antioxidant Activity of Essential Oils from Populations of Baccharis dracunculifolia DC. in Southern Brazil

HIGHLIGHTS Essential oils from populations of B. dracunculifolia were investigated. β-pinene and (E)-nerolidol were the main compounds in B. dracunculifolia populations. The difference in the chemical profile of the essential oils is quantitative only. There is a negative correlation between the antioxidant activity and spathulenol.

Abstract Baccharis dracunculifolia DC. is a Brazilian native plant, presenting wide chemical diversity and numerous pharmaceutical and industrial applications. This research assessed the yield, antioxidant activity and the chemical similarity of essential oils from 10 populations of B. dracunculifolia in the state of Paraná, southern Brazil. The extraction of the volatile compounds was carried out by hydrodistillation, the chemical composition was determined by GC/FID and GC/MS and the antioxidant activity by the DPPH method. The essential oil yield of wild B. dracunculifolia populations ranged from 0.14 to 0.87%. The oils were predominantly composed of oxygenated sesquiterpenes (34.16 - 51.01%), monoterpene hydrocarbons (18.02 - 46.17%) and sesquiterpenes hydrocarbons (9.60 - 17.70%). The major compounds found in all populations were β-pinene (7.65 - 29.8%) and (E)-nerolidol (9.11 - 21.68%). Essential oil solutions (20%) from different populations presented antioxidant capacity ranging from 27.78 to 91.67%. A negative correlation was found between the antioxidant activity and spathulenol (r = -0.696). Multivariate analyses separated the populations into three groups: (1) low concentrations of α-pinene (2.02 - 2.06%), (2) high concentrations of α-pinene (4.17 - 4.61%) and β-pinene (22.54 - 29.80%), and (3) intermediate concentrations of α-pinene (2.38 - 3.31%), β-pinene (12.77 - 19.03%) and spathulenol (6.02 - 9.06%).

The anticonvulsant effects of Ducrosia anethifolia (Boiss) essential oil are produced by its main component alpha-pinene in rats / Os efeitos anticonvulsivantes do óleo essencial de Ducrosia anethifolia (Boiss) são realizados pelo seu principal componente alfa-pineno em ratos

ABSTRACT Ducrosia anethifolia has been recommended as a remedy for neurological disorders. However, the anticonvulsant effects of D. anethifolia essential oil (DAEO) and its major constituent α-pinene have not yet been clarified. Methods: A rat model of pentylenetetrazole (PTZ)-induced convulsions was used. Oxidant and antioxidant parameters were assayed in the temporal lobe. Results: The data showed that DAEO (50, 100 and 200 mg/kg, i.p.) and α-pinene (0.2 and 0.4 mg/kg i.p.) delayed the initiation time, and reduced the duration of myoclonic and tonic-clonic seizures following PTZ injection. The PTZ produced oxidative stress so that malondialdehyde and hydrogen peroxide levels were increased and catalase and peroxidase activity decreased. Pretreatment with DAEO and α-pinene significantly inhibited the above-mentioned enzymatic changes in PTZ-treated animals. Conclusion: The results suggest that α-pinene, at teast in part, was responsible for the induction of the anticonvulsant and antioxidant effects of DAEO in rats.

RESUMO A Ducrosia anethifolia tem sido recomendada como remédio para os distúrbios neurológicos. No entanto, os efeitos anticonvulsivantes do óleo essencial de Ducrosia anethifolia (DAEO) e do seu principal constituinte atfa-pineno (α-pineno) ainda não foram clarificados. Métodos: Foi utilizado um modelo de rato de convulsões induzidas por pentilenotetrazol (PTZ). Os parâmetros oxidante e antioxidante foram ensaiados no lobo temporal do cérebro. Resultados: Os dados mostraram que DAEO (50, 100 e 200 mg / kg, i.p.) e α-pineno (0,2 e 0,4 mg / kg i.p.) retardaram o tempo de iniciação e reduziram a duração das crises mioclônicas e tônico-clônicas após a injeção de PTZ. O PTZ produziu estresse oxidativo, de modo que os níveis de malondialdeído (MDA) e de peróxido de hidrogênio aumentaram e a atividade da catalase e da peroxidase diminuiu. O pré-tratamento com DAEO e α-pineno inibiu significativamente as alterações enzimáticas mencionadas em animais tratados com PTZ. Conclusão: O resultado sugere que α-pineno, peto menos em parte, é responsável peta indução dos efeitos anticonvulsivantes e antioxidantes da DAEO em ratos.

Feasibility of vapor permeation technology for separating of Asari Radix et Rhizoma essential oil-bearing water / 中草药

Objective: To verify the feasibility of vapor-permeable membrane technology for the separation of water bodies containing essential oil of Asari Radix et Rhizoma (ARR) essential oil, and then to apply vapor permeate technology to the separation of more essential oils of traditional Chinese medicine. Methods: The polydimethylsiloxane/polyvinylidene fluoride (PDMS/PVDF) composite flat membrane and polyvinylidene fluoride (PVDF) flat membrane were collected as the membrane material. The oil-bearing water body of ARR volatile oil was separated by vapor permeate technology, and the oil penetration rate of two kinds of membranes was calculated. At the same time, the changes of the composition and content of the essential oil before and after the membrane were analyzed by gas chromatography-mass spectrometry (GC-MS). Results: The results showed that the essential oil penetration rate was significantly higher than that of PDMS/PVDF membrane when PVDF membrane was used as membrane material. GC-MS qualitative analysis results showed that the composition of the essential oil in the penetrants of the two membranes was basically the same as that of the essential oil obtained by the traditional steam distillation method. The content of α-pinene, β-pinene, 3,5-dimethoxytoluene, and methyl eugenol were determined by double internal standard method. The results showed that the content of each component in the PVDF membrane permeation was significantly higher than that of the PDMS/PVDF membrane permeation solution. Conclusion: It is feasible to separate the oil containing water from the essential oil of ARR by vapor permeation membrane technology. Compared with the PDMS/PVDF membrane, the PVDF membrane is more suitable for separating the oil containing water of the essential oil of ARR.

Simultaneous Determination of Contents of 6 Components in the Oil of Blumea balsamifera by GC / 中国药房

OBJECTIVE： To establish a method for simultaneous determination of contents of β-pinene， linalool， L-camphor， L-borneol， β-caryophyllene and xanthoxylin in the oil of Blumea balsamifera. METHODS： GC method was adopted. The determination was performed on RTX-1701 capillary column （programmed temperature）. The FID detector was controlled at 240 ℃. The inlet temperature was set at 240 ℃. The carrier gas was high-purity nitrogen 3 mL/min. The the sample size was 0.5 μL， and split ratio was 50 ∶ 1. RESULTS： The linear range of β-pinene， linalool， L-camphor， L-borneol， β-caryophyllene and xanthoxylin were 0.029 7-0.267 1 mg/mL （r＝0.999 9）， 0.024 3-0.218 9 mg/mL （r＝0.999 9）， 0.126 0-1.134 0 mg/mL （r＝0.999 9）， 0.217 2-1.954 8 mg/mL （r＝0.999 9）， 0.136 3-1.226 9 mg/mL （r＝0.999 9）， 0.044 5-0.400 3 mg/mL（r＝0.999 5）， respectively. The limits of quantitation were 0.028 5， 0.008 7， 0.018 6， 0.016 8， 0.014 5， 0.042 1 mg/mL； the limits of detection were 0.009 4， 0.002 9， 0.006 1， 0.005 5， 0.004 8， 0.013 9 mg/mL， respectively. RSDs of precision， stability， reproducibility and durability tests were all lower than 3％. The average recoveries were 98.13％-101.30％（RSD＝1.20％，n＝9），98.44％-101.81％（RSD＝1.28％，n＝9），98.26％-101.05％（RSD＝1.19％，n＝9），99.08％-101.58％（RSD＝0.89％，n＝9），98.66％-101.66％（RSD＝1.17％，n＝9），98.84％-103.60％（RSD＝0.96％，n＝9）， respectively. The contents of 6 components in the sample were 14.552-46.766， 16.951-22.096， 80.597-113.115， 205.224-242.537， 47.761-135.697， 26.493-45.771 mg/g， respectively. CONCLUSIONS： The established method is simple， accurate， precise and reproducible， which can be used for simultaneous determination of contents of 6 components in the oil of B. balsamifera. It can provide reference for comprehensive evaluation and extraction technology study of the oil of B. balsamifera.

Optimization of the Extraction Technology of Volatile Oil from Schizonepeta tenuifolia and Forsythia suspensa / 中国药房

OBJECTIVE： To establish the method for content determination of volatile oil of Schizonepeta tenuifolia and Forsythia suspensa， and to optimize the extraction technology of the volatile oil. METHODS： The contents of β-pinene and pulegone were determined by GC method. The determination was performed on Hp-5 capillary column. The detector was hydrogen flame ion detector with programmed temperature. The sample size was 0.5 μL， the split ratio was 70 ∶ 1， the carrier gas was nitrogen， the inlet temperature was 250 ℃， the detector temperature was 280 ℃， the air flow rate was 390 mL/min， the hydrogen flow rate was 36 mL/min， the tail flow rate was 15 mL/min， and the nitrogen flow rate was 1 mL/min. Based on single factor test， orthogonal test combined with information entropy method were used to optimize the extraction technology of S. tenuifolia and F. suspensa using soaking time， extraction time， material-liquid ratio and forsythia grain size as factors， with the extraction amount of volatile oil， the content of β-pinene and pulegone and their comprehensive score as indexes.  RESULTS： The linear range of β-pinene and pulegone 1.575-7.875（r＝0.999 9） and 1.892-9.46 μg（r＝0.999 7）， respectively. The limits of quantitation were 0.10 and 0.25 μg； the limits of detection were 0.03 and 0.08 μg； RSDs of precision， stability and reproducibility tests were less than 2％ （n＝6）； the recoveries were 97.77％-100.01％ （RSD＝0.93％，n＝9） and 96.47％-99.00％（RSD＝0.89％， n＝9）. The optimal extraction technology was soaking 2 h， extracting for 6 h， 10-fold water （mL/g）， half a clove of granularity. Under this condition， the extraction amount of volatile oil， the contents of β-pinene and pulegone were 3.6 mL， 1 450.4 mg， 127.6 mg， respectively. RSD were 1.62％， 0.20％， 1.42％. CONCLUSIONS： Established method is simple， accurate and reproducible， and the optimal extraction technology is stable and feasible.

Phytochemical and Antimicrobial Evaluation of the Essential Oil of Croatian Salvia brachyodon Vandas

Backgraund: This study was designed to evaluate the phytochemical profile and the antimicrobial potential of the essential oil of Salvia brachyodon Vandas growing wild in Croatia. Short tooth sage (S. brachyodon Vandas),an endemic species that grows in the south east areas of the Adriatic coast in Croatia, Bosnia and Herzegovina, and Montenegro. It is a perennial plant that grows up to 70-80 cm in height and flowers from July to September. Methodology: The phytochemical components of the essential oil were identified by gas chromatography-mass spectrometry (GC-MS) analysis. The antimicrobial activity was assessed against a panel of representative Gram-positive and Gram-negative bacteria as well as fungi. The antimicrobial activities of the oil against pathogenic microorganisms were determined by using agar disc diffusion and broth microdilution methods. Results: From the thirty-eight identified constituents representing 95.7% of the oil, 1,8-cineole (16.7%), β-pinene (19.7%) and α-pinene (7.6%), were the major components. The levels of oxygenated monoterpenes such as camphor (5.6%), borneol (4.2%), myrtenol (2.4 %) and terpinen-4-ol were significant. Other important compounds were sesquiterpenes hydrocarbons β-caryophyllene (6.6%), α-humulene (4.9%), viridiflorol (3.0%), spathulenol (2.9%) and aromadendrene. Preliminary antimicrobial screening revealed that the oil exhibited a very interesting antimicrobial profile. The oil exhibited moderate in vitro antibacterial activity after it was tested against twenty pathogenic bacteria and fungal strains, but high antimicrobial activity observed against medically important pathogens such as E. coli O157:H7, Listeria monocytogenes and Candida albicans. Conclusion: Results presented here may suggest that the essential oil of S. brachyodon possess antimicrobial properties, and is, therefore, a potential source of antimicrobial ingredient in food and pharmaceutical industry. The obtained results are preliminary and a further research is needed in order to obtain information regarding the practical effectiveness of essential oil to prevent the growth of foodborne and spoilage microbes under specific application conditions.

Evaluation of the chemical composition and variability of the volatile oils from Trembleya parviflora leaves

Abstract Trembleya parviflora (D. Don) Cogn., Melastomataceae, also known as "quaresmeira-branca", is a subshrub that is commonly used to treat verminosis, scabies, dermatoses, rheumatism, vaginal infections, ulcerations and wounds. The aim of this work was to perform a morphological study of T. parviflora, evaluate the composition and chemical variability of the volatile oils from the leaves, perform phytochemical screening of the powder from the leaves and to define parameters for quality control of the plant material. Macroscopic characterization of T. parviflora was carried out by naked eye in Serra dos Pireneus, Pirenópolis, Goiás for 12 months. Volatile oils were subjected to hydrodistillation with Clevenger apparatus and analyzed by gas chromatography-mass spectrometry. Phytochemical screening and ash and volatile compound content determination were performed by conventional techniques. T. parviflora has simple, oppositely crossed and petiolate leaves. The inflorescence of this plant is a cyme. The presence of coumarins, steroids, triterpenes, flavonoids and tannins was observed. The total ash content was 4.05 ± 0.02%; the insoluble ash content was 0.10 ± 0.03%; and the volatile compound content was 9.53 ± 0.02%. The major compounds present in the volatile oils were α-terpineol (2.7-16.5%), α-pinene (0.6-25.4%), β-pinene (2.7-23.1%), sabinene (1.2-14.1%), acetoxyeudesman-4-α-ol (0.6-6.3%) and 2,4a-8,8-tetramethyldecahydrocyclopropanaphtalene (2.4-24.4). Two clusters were identified: Cluster I represented the period with low levels of rainfall, and Cluster II represented the period with high levels of rainfall. This study provides data that can be applied for the quality control of powdered leaves and is the first description of the chemical composition and variability of the volatile oils from the leaves of T. parviflora.

Composición química del aceite esencial de las hojas de Hedyosmum luteynii Todzia (Chloranthaceae)

El objetivo de este trabajo fue la caracterización química del aceite esencial de Hedyosmum luteynii, a partir de muestras recolectadas en el bosque natural Jacarón, cantón Colta, provincia de Chimborazo, Ecuador. El aceite esencial se extrajo por hidrodestilación; el análisis de la composicion se realizó mediante un cromatógrafo de gases acoplado a un espectrómetro de masas; la identificación de los componentes se realizó por comparación de sus espectros de masas y de los índices de Kováts reportados en la literatura. Se identificaron 28 compuestos, correspondientes al 98.62% del total de los constituyentes, siendo la mayoría monoterpenos hidrocarburos naturales (83.21%). Los constituyentes más abundantes fueron: α-felandreno (32.72%), α-pineno (13.20%), (Z)-β-ocimeno (10.99%), silvestreno (6.51%), biciclogermacreno (5.05%), 1.8-cineol (4.95%), (E)-β-ocimeno (3.88%) y germacreno D (3.20%). Es la primera vez que se reporta al silvestreno como un componente importante en el aceite esencial de una de las especies del genero Hedyosmum. Este resultado hace evidente una marcada diferencia en la composición química del aceite esencial de H. luteynii respecto a otras especies del género Hedyosmum.

In this work, essential oil of Hedyosmum luteynii is characterize from samples collected in the Jacarón natural forest, Colta, Chimborazo, Ecuador. The essential oil was extracted by hydrodistillation; analysis of the composition was carried out by gas chromatograph, coupled to a mass spectrometer; identification of the components was made by comparing their mass spectra and the Kováts indexes reported in the literature. Twenty-eight compounds were identified, 98.62% of the total components. Natural hydrocarbon monoterpenes were the highger (83.21%). The most abundant components were: α-phellandrene (32.72%), α-pinene (13.20%), (Z)-β-ocimene (10.99%), silvestrene (6.51%), bicyclogermacrene (5.05%), 1.8-cineol (4.95%), (E)-β-ocimene (3.88%) and germacrene D (3.20%). For first time, silvestrene is reported as an important component of essential oil in one of the species of the genus Hedyosmum. This result is a important difference in the chemical composition of the essential oil of H. luteynii respect to other species of Hedyosmum.

Synthesis, biological activity, computer aided drug design of alpha-pinene derivatives / 中国中药杂志

Based on the anticancer mechanism of biological alkylating agent, we designed and synthesized two alpha pinene derivatives:(1R,5S)-(6,6-dimethylbicyclo[3,1,1]hept-2-en-2-yl)methyl benzenesulfonate and (1R,5S)-(6,6-dimethylbicyclo[3,1,1]hept-2-en-2-yl)methyl 4-methylbenzenesulfonate, of which structures were confirmed by ¹H-NMR, HPLC and MS date. These two compounds showed a good inhibition of tumor cells' proliferation. Further, the computer siuulation of molecular docking and metabolic kinetics indicated that these two copounds may have stable molecular complexation with protein CDK2, which closely related to the cell cycle.

Simultaneous Determination of α-pinene,β-pinene and Linalool in Volatile Oil from Hedychium flavum by GC / 中国药房

OBJECTIVE:To establish a method for simultaneous determination of α-pinene,β-pinene and linalool in volatile oil of Hedychium flavum. METHODS:The volatile oil was extracted from H. flavum according to steam distillation stated in Chinese Pharmacopoeia(2015 edition volume Ⅳ). GC method was adopted to determine the content. The determination was performed on HP-5 capillary chromatographic column(30 m×0.032 mm×0.25 μm)with hydrogen flame ion detector(FID),nitrogen gas as carrier gas,with injector temperature of 200 ℃,and detector temperature of 250 ℃,column flow rate of 0.8 mL/min, samples size of 1 μL and split ratio of 40:1 by programmed temperature. RESULTS:The linear range of α-pinene,β-pinene and linalool were 0.090 5-2.413 3 mg/mL(r=0.999 9),0.098 3-2.620 0 mg/mL(r=0.999 9),0.169 1-4.510 0 mg/mL(r=0.999 8), respectively. RSDs of precision,stability(12 h),reproducibility tests were no more than 2.0%(n=6). The average recoveries were 99.84%(RSD=0.49%,n=6),100.24%(RSD=1.38%,n=6),99.41%(RSD=1.67%,n=6),respectively.The contents of α-pinene,β-pinene and linalool ranged 0.214 4-1.325 0,0.766 2-3.172 1,0.357 4-1.518 7 mg/g in volatile oil from 23 batches of H. flavum produced in Guizhou province. CONCLUSIONS:The method established in this experiment is rapid,simple and accurate.It can be used for the content determination of α-pinene,β-pinene and linalool in volatile oil from H.flavum.

Anti-cariogenic Properties of α-Pinene, a Monoterpene in Plant Essential Oil

Dental caries is the most common chronic disease in the dental field. Streptococcus mutans (S. mutans) is the most important bacteria in the formation of dental plaque and dental caries. In a previous study, we confirmed that the essential oil of Chrysanthemum boreale has antibacterial activity against S. mutans. Alpha-pinene is one of the major chemical components of Chrysanthemum boreale essential oil. In the present study, we investigated the inhibitory effects of α-pinene on cariogenic properties such as growth, acid production, biofilm formation, and bactericidal activity on S. mutans. Alpha-pinene at a concentration range of 0.25-0.5 mg/mL significantly inhibited the growth of S. mutans and acid production of S. mutans. Biofilm formation was significantly inhibited at < 0.0625 mg/mL α-pinene, similar to the data from scanning electronic microscopy. Under confocal laser scanning microscopy, the bacterial viability was decreased by α-pinene in a dose-dependent manner. These results suggested that α-pinene may be a useful agent for inhibiting the cariogenic properties of S. mutans.

Comparative Analysis on Simultaneous Determination of α-pinene, β-pinene, Eucalyptol and Linalool in Baeckea Frutescens Preparations by Reference Substances Method and Reference Extract Method / 中国中医药信息杂志

Objective To determinate the contents of α-pinene, β-pinene, eucalyptol and linalool in Baeckea frutescens by reference substances method and reference extract method respectively; To explore the feasibility of replacing single component reference by control extracts in assay. Methods The GC system consisted of a quartz column DB-5 (60 m×0.25 mm×0.25 μm); The temperature programming rose from 80 ℃ (15 min) to 90 ℃ by 1 ℃/min, lasting 2 min, then 10 ℃/min to 110 ℃, then 25 ℃/min to 240 ℃, lasting 8 min in the end; The temperature of the entrance of capillary vessel column was 250 ℃, and the temperature of the detector was 250 ℃. Results α-pinene, β-pinene, eucalyptol and linalool were in a good linear relationship within each concentration scope (r≥0.999). The average recovery rates were in the range of 96.5%–102.2%. The results of t-test demonstrated that there is no significant difference between the two methods. Conclusion The reference extract method can be used as a quality evaluation pattern for Baeckea frutescens.

Curative effect of Eucalyptol on the treatment of chronic rhinosinusitis / 中国耳鼻咽喉头颈外科

OBJECTIVE To explore the curative effect of Eucalyptol on the treatment of chronic rhinosinusitis (CRS). METHODS CRS patients were randomly divided into treatment group (conventional medications plus Eeucalyptol) and control group (conventional medications). The therapeutic effect of Eucalyptol were objectively and subjectively evaluated after treatment. RESULTS Evaluation at 3, 6 and 12 month after treatment, we found that the curative effect of the treatment group was better than that of the control group. There was a significant difference. CONCLUSION Eucalyptol as a kind of mucus dissolving medication, it has good treatment effect for CRS and is worth popularizing.

Effects of α-pinene extracted from pine needle on expression of miR-221 and its potential target genes in human hepatocellular carcinoma cells / 中国中药杂志

To investigate the anti-hepatoma mechanism of α-pinene, HepG2 cell was treated with α-pinene and the change of cell cycle was examined by flow cytometry. The expression of miR-221, which was related the regulation of G₂/M phase, was detected by quantitative Real-time PCR. Meanwhile, TargetScan and other online bioinformatics methods were used to analyze and predict the target genes of miR-221, then the expression level of related target genes were detected by quantitative Real-time PCR. The results showed that α-pinene inhibited the proliferation of HepG2 cells in dose-dependent manner. It was also proved that HepG2 cells were arrested at G₂/M phase by α-pinene (P<0.05). The expression of miR-221 was down-regulated in α-pinene treated HepG2 cell. The bioinformatics analysis showed that CDKN1B/P27 and CDKN1C/P57 may be the protential targets of miR-221 and both of them were significantly up-regulated（P<0.001,P<0.05）by α-pinene treatment. According to these results, it was believed that α-pinene may inhibit the proliferation of hepatocellular carcinoma cells through arrest the cell at G₂/M phase, which may be associated with the down-regulate of the miR-221 expression and up-regulate of the CDKN1B/P27 and CDKN1C/P57 expression.

Composition of the essential oil from leaves of Smallanthus quichensis (Asteraceae) from Costa Rica / Composición química del aceite esencial de las hojas de Smallanthus quichensis (Asteraceae) de Costa Rica

Smallanthus is a genus of flowering plants in the Asteraceae family, which has about 24 species, ranging mostly from southern Mexico and Central America to the Andes in South America. The aim of the present study was to identify the chemical composition of leaf essential oil of S. quichensis, growing wild in Costa Rica. The extraction of the oils was carried out by the hydrodistillation method, using a modified Clevenger type apparatus. The chemical composition of the oils was analyzed by capillary GC-FID and GC-MS using the retention indices on DB-5 type capillary column. A total of 100 compounds were identified, accounting for about 90 percent of the total amount of the oils. Smallanthus quichensis leaf produced a monoterpenoid-rich oil, whose composition was dominated by αlfa-pinene (64.5 percent) and 1,8-cineole (9.7 percent) or, in a different sample, by αlfa-pinene (35.5 percent) with moderate amounts of p-cymene (11.5 percent), beta-phellandrene (9.2 percent), alfa-phellandrene (9.0 percent) and limonene (5.8 percent). This is the first report of the chemical composition of the essential oil obtained from this plant species.

Smallanthus es un género de plantas perteneciente a la familia Asteraceae que contiene aproximadamente 24 especies, la mayoría ubicadas desde el sur de México, América Central y hasta la cordillera de los Andes en América del Sur. El objetivo del presente estudio fue el de identificar la composición química del aceite esencial de las hojas de S. quichensis. La extracción se realizó mediante el método de hidrodestilación, empleando un instrumento de tipo Clevenger. Se analizó la composición de los aceites mediante CG-FID y CG-EM, utilizando índices de retención obtenidos en una columna capilar tipo DB-5. Se identificaron 100 compuestos, correspondientes a un 90 por ciento de los constituyentes totales. Los aceites están constituidos principalmente por monoterpenoides (ca. 85 por ciento). Los componentes mayoritarios se identificaron como alfa-pineno (64.5 por ciento), acompañado por 1,8-cineol (9.7 por ciento) o, en otra muestra distinta, αlfa-pineno (35.5 por ciento) junto a cantidades moderadas de p-cimeno (11.5 por ciento), beta-felandreno (9.2 por ciento), alfa-felandreno (9.0 por ciento) y limoneno (5.8 por ciento). Este es el primer informe acerca de la composición química de aceites esenciales obtenidos de S. quichensis.

Chemical composition and antibacterial activities of seven Eucalyptus species essential oils leaves

BACKGROUND: In this paper, we have studied the essential oils chemical composition of the leaves of seven Eucalyptus species developed in Tunisia. Eucalyptus leaves were picked from trees growing in different arboretums in Tunisia. Choucha and Mrifeg arboretums located in Sedjnene, region of Bizerte (Choucha: E. maideni, E. astrengens et E. cinerea; Mrifeg : E. leucoxylon), Korbous arboretums located in the region of Nabeul, North East Tunisia with sub-humid bioclimate, (E. lehmani), Souiniet-Ain Drahem arboretum located in region of Jendouba (E. sideroxylon, E. bicostata). Essential oils were individually tested against a large panel of microorganisms includingStaphylococcus aureus (ATCC 6539), Escherichia coli (ATCC 25922), Enterococcus faecalis (ATCC29212), Listeria ivanovii (RBL 30), Bacillus cereus (ATCC11778). RESULTS: The yield of essential oils ranged from 1.2% to 3% (w/w) for the different Eucalyptus species. All essential oils contain α-pinene, 1,8-cineol and pinocarveol-trans for all Eucalyptus species studied. The 1,8-cineol was the major compound in all species (49.07 to 83.59%). Diameter of inhibition zone of essential oils of Eucalyptus species varied from 10 to 29 mm. The largest zone of inhibition was obtained for Bacillus cereus (E. astrengens) and the lowest for Staphylococcus aureus (E. cinerea). The essential oils from E. maideni, E. astrengens, E. cinerea (arboretum of Bizerte), E. bicostata(arboretum of Aindraham) showed the highest antibacterial activity against Listeria ivanovii and Bacillus cereus. CONCLUSION: The major constituents of Eucalyptus leaves essential oils are 1,8-cineol (49.07 to 83.59%) and α-pinene (1.27 to 26.35%). The essential oils from E. maideni, E. astrengens, E. cinerea, E. bicostatashowed the highest antibacterial activity against Listeria ivanovii and Bacillus cereus, they may have potential applications in food and pharmaceutical products.

Effectiveness of mucolytic agent treating with chronic rhinosinusitis:a randomized clinical research / 中国基层医药

Objective To investigate the effect of mucolytic agents during endoscopic sinus surgery.Methods 84 patients of chronic rhinosinusitis were employed in the study,which were randomly divided into the control group(41 subjects) and study group(43 subjects) according to the digital table.The patients in the control group adopted regular therapies,and study group were received regular therapies as well as mucolytic agents(eucalyptol-limonene-pinene soft capsule,300 mg,three times a day) application for 8 weeks.Followed visiting lasted for 6 months.The VAS scores,Lund-Mackay scores and efficient rate were compared.Results The VAS scores and Lund -Mackay scores in the control group after treatment were (3.63 ± 1.71)points and (8.34 ±4.68) points,and which in study group were (2.03 ± 1.67) points and (6.48 ± 3.59) points,and the difference was statistic significant (t =4.338 and t =2.049,P ＜ 0.05).According to CPOS-2012,in the control group,19 cases were controlled,14 cases were partly controlled,and 8 cases were uncontrolled,the whole efficient rate was 80.5％.in the study group,24 cases were controlled,17 cases were partly controlled,and 2 cases were uncontrolled,the whole efficient rate was 95.3％.Compared to the control group,the efficient rate of the study group was higher,the result showed statistical significance (x2 =4.420,P ＜ 0.05).Conclusion Using mucolytic agent after surgery can improve the controlled rate and affect the success of treatment of chronic rhinosinusitis.

Biotransformation of (-)-alpha-Pinene by Whole Cells of White Rot Fungi, Ceriporia sp. ZLY-2010 and Stereum hirsutum

Two white rot fungi, Ceriporia sp. ZLY-2010 (CER) and Stereum hirsutum (STH) were used as biocatalysts for the biotransformation of (-)-alpha-pinene. After 96 hr, CER converted the bicyclic monoterpene hydrocarbon (-)-alpha-pinene into alpha-terpineol (yield, 0.05 g/L), a monocyclic monoterpene alcohol, in addition to, other minor products. Using STH, verbenone was identified as the major biotransformed product, and minor products were myrtenol, camphor, and isopinocarveol. We did not observe any inhibitory effects of substrate or transformed products on mycelial growth of the fungi. The activities of fungal manganese-dependent peroxidase and laccase were monitored for 15 days to determine the enzymatic pathways related to the biotransformation of (-)-alpha-pinene. We concluded that a complex of enzymes, including intra- and extracellular enzymes, were involved in terpenoid biotransformation by white rot fungi.