Transcriptome differences between 20- and 3, 000-year-old Platycladus orientalis reveal that ROS are involved in senescence regulation

Background: Platycladus orientalis has an extremely long life span of several thousands of years, attracting great interests in the mechanisms involved in such successful senescence regulation and resistance at physiological and molecular levels. Results: The levels of reactive oxygen species (ROS) were higher in 3,000-year-old than in 20-year-old P. orientalis, and the activities of GR and GSH demonstrated the same trend. We produced and analyzed massive sequence information from pooled samples of P. orientalis through transcriptome sequencing, which generated 51,664 unigenes with an average length of 475 bp. We then used RNA-seq analysis to obtain a high-resolution age­course profile of gene expression in 20- and 3,000-year-old P. orientalis individuals. Totally, 106 differentially expressed genes were obtained, of which 47 genes were downregulated and 59 upregulated in the old tree. These genes were involved in transcription factors, hormone-related responses, ROS scavengers, senescence-related responses, stress response, and defense and possibly play crucial roles in tackling various stresses in the 3,000-year-old P. orientalis during its life time. The expression patterns of genes related to ROS homeostasis further indicated that the high ability of ROS scavenging could be helpful for the 3,000-year-old P. orientalis to resist senescence. Conclusions: This study provides a foundation for the elucidation of senescence resistance through molecular studies and the discovery of useful genes in P. orientalis.

Isolation of high-quality RNA from Platycladus orientalis and other Cupressaceae plants

Platycladus orientalis has a lifespan of several thousand years in China, making it a good plant in which to study aging at the molecular level, but this requires sufficient quantities of high-quality P. orientalis RNA. However, no appropriate methods have been reported for total RNA isolation from P. orientalis leaves. The TRIzol method did not extract RNA, while cetyltrimethylammonium bromide, sodium dodecyl sulfate-phenol, and plant RNAout kit (Tianz, Inc., China) protocols resulted in low yields of poor quality RNA. Isolating total RNA using the Spectrum™ Plant Total RNA Kit (Sigma-Aldrich, St. Louis, MO, USA) resulted in a high-quality product but a low yield. However, the two-step removal of polyphenols and polysaccharides in the improved plant RNAout kit protocol resulted in the isolation of RNA with a 28S:18S rRNA ratio of band intensities that was ~2:1, the A260/A280 absorbance ratio was 2.03, and the total RNA yield from P. orientalis leaves was high. This protocol was tested on different P. orientalis tissues of different ages and on leaves of five other Cupressaceae plants. The total RNAs were successfully used in complementary DNA synthesis for transcriptome sequencing and would be suitable to use in additional experiments. The results of this study will benefit future studies in Cupressaceae plants.

Anti-listerial synergism of leaf essential oil of Metasequoia glyptostroboides with nisin in whole, low and skim milks

OBJECTIVE@#To examine the individual and synergistic anti-listerial effect of nisin and leaf essential oil of Metasequoia glyptostroboides (M. glyptostroboides) against one of the leading foodborne pathogens Listeria monocytogenes (L. monocytogenes) ATCC 19116 in milk samples.@*METHODS@#The whole (8%), low (1%) and skim (no fat content) milk samples were inoculated with L. monocytogenes ATCC 19116 along with leaf essential oil of M. glyptostroboides or nisin alone as well in combinations.@*RESULTS@#In this study, the leaf essential oil at the concentrations of 2% and 5% revealed strong anti-listerial effect against L. monocytogenes ATCC 19116 in all categories of milk samples. Nisin at the concentrations of 250 and 500 IU/mL displayed a strong inhibitory effect against ATCC 19116 as compared to the control group. Additionally, synergistic combinations of leaf essential oil (1%) and nisin (62.5, 125, 250 and 500 IU/mL) also had a remarkable anti-listerial synergism in all the tested milk samples including whole, low and skim milk after 14 days.@*CONCLUSIONS@#As a major finding, the leaf essential oil of M. glyptostroboides might be a useful candidate for using in food industry to control the growth of foodborne pathogenic bacteria as confirmed by its potent anti-listerial synergistic effect with nisin against L. monocytogenes ATCC 19116 in different milk samples.

Antioxidant, lipid peroxidation inhibition and free radical scavenging efficacy of a diterpenoid compound sugiol isolated from Metasequoia glyptostroboides

OBJECTIVE@#To investigate the antioxidant efficacy of a biologically active diterpenoid compound sugiol isolated from Metasequoia glyptostroboides (M. glyptostroboides) in various antioxidant models.@*METHODS@#An abietane type diterpenoid sugiol, isolated from ethyl acetate extract of M. glyptostroboides cones, was analyzed for its antioxidant efficacy as reducing power ability and lipid peroxidation inhibition as well as its ability to scavenge free radicals such as 1,1-diphenyl-2-picryl hydrazyl, nitric oxide, superoxide and hydroxyl radicals.@*RESULTS@#The sugiol showed significant and concentration-dependent antioxidant and free radical scavenging activities. Consequently, the sugiol exerted lipid peroxidation inhibitory effect by 76.5% as compared to α-tocopherol (80.13%) and butylated hydroxyanisole (76.59%). In addition, the sugiol had significant scavenging activities of 1,1-diphenyl-2-picryl hydrazyl, nitric oxide, superoxide and hydroxyl free radicals in a concentration-dependent manner by 78.83%, 72.42%, 72.99% and 85.04%, when compared to the standard compound ascorbic acid (81.69%, 74.62%, 73.00% and 73.79%) and α-tocopherol/butylated hydroxyanisole (84.09%, 78.61%, 74.45% and 70.02%), respectively.@*CONCLUSIONS@#These findings justify the biological and traditional uses of M. glyptostroboides or its secondary metabolites as confirmed by its promising antioxidant efficacy.

Filtration of active fractions with hemostasis effect from platycladi cacumen carbonisatum / 中国中药杂志

<p><b>OBJECTIVE</b>To discuss the hemostasis of the different polarities of Platycladi Cacumen Carbonisatum (PCC) on the blood heat and hemorrhage syndrome rat model induced by dry yeast.</p><p><b>METHOD</b>The SD rats were divided into seven groups. Yunnan Baiyao was taken as the positive control drug. The rats in the control group and model group were fed with CMC-Na for 7 days, and the rats in other groups were fed with corresponding drugs simultaneously. On day 7, the blood heat and hemorrhage syndrome rat model was established. Indexes including the whole blood viscosity, plasma viscosity, thrombin time (TT), activated partial thromboplastin time (APTT), prothrombin time (PT), fibrinogen content (FIB), red blood cell (RBC), hemoglobin (HGB), hematocrit (HCT), blood platelet count (PLT), thrombocytocrit (PCT), mean platelet volume (MPV), platelet distribution width (PDW) and the rate of platelet aggregation induced by ADP were detected. Additionally, the pathological examinations of lungs among each group were compared.</p><p><b>RESULT</b>Compared with the control group, the RBC, HGB and HCT of rats in the model group increased significantly, with distinct increase in high, middle and low whole blood viscosity and plasma viscosity of rats in the model group; TT and APTT were notably prolonged, while PT was notably shortened, with significant increase in FIB content; PLT, PCT, MPV and PDW remarkably increased; Additionally, the rate of platelet aggregation induced by ADP significantly decreased. After ig administration of the ethyl acetate extract of PCC, the low whole blood viscosity and plasma viscosity remarkably decreased; TT and APTT were significantly shortened, with notable reduction in PDW and in FIB content Additionally, the rate of platelet aggregation induced by ADP significantly increased. The injury of lungs was also improved in ethyl acetate extract group. The rate of platelet aggregation induced by ADP of n-butanol extract group notablly increased. Plasma viscosity of water extract group remarkably decreased, with TT being significantly shortened. But the effects of n-butanol extract or water extract were weaker than that of ethyl acetate extract. And the effect of petroleum ether extract was the weakest.</p><p><b>CONCLUSION</b>Ethyl acetate extract is the active part of PCC, showing the effect of hemostasis by reducing the low whole blood and plasma viscosity, improving coagulation function mainly by acting on the endogenous coagulation, and ameliorating the function of platelet aggregation.</p>

Purification of polyphenols from Sabina vulgaris antoine and its antioxidant properties / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To purify polyphenols from Sabina vulgaris and to investigate its antioxidant properties.</p><p><b>METHODS</b>Polyphenols were purified from Sabina vulgaris Antoine with macroporous resin HPD-700, and the quantity of polyphenols was determined by Folin-Ciocalteu colorimetry. The antioxidant properties of polyphenols were evaluated by total antioxidant capacity (T-AOC) and its activities of scavenging DPPH (1,1 diphenyl-2-picry-hydrazyl) radicals, superoxide anion (O2·-), hydroxyl free radicals (OH·) and ferric reducing antioxidant power (FRAP).</p><p><b>RESULTS</b>After purification, the purity of polyphenols increased from 0.053% to 0.995%.The antioxidant properties study showed that its inhibition rate of scavenging DPPH radicals and FRAP was 151.83 U/ml and 204.59 U/ml. Its scavenging capacity for superoxide anion (O2·-) and hydroxyl free radicals (OH·) was 151.83 U/ml and 204.59 U/ml. The total antioxidant capacity was 72.68 U/ml.</p><p><b>CONCLUSION</b>Polyphenols from Sabina vulgaris Antoine have high antioxidant properties, suggesting that it worth further study of its pharmacological effects.</p>

DNA sequence variation of drought-response candidate genes in Austrocedrus chilensis

Background: Austrocedrus chilensis (D. Don) Pic. Ser. et Bizzarri commonly known as Patagonian cypress is a member of the Cupressaceae family, characterized by a high adaptive potential for growing in marginal areas and good timber quality. The species grows over a wide area and under a wide range of rainfall. This study assessed adaptive genetic variation at SNP level in candidate genes involved in response to drought stress. Results: A total of 18 single nucleotide polymorphisms (SNPs) were found among 1,428 bp. Average nucleotide diversity value (π = 0.00312) was similar to those previously reported in other Cupressaceae. The Fst average among genes and populations was 0.163 and the lowest differentiation was observed in continuous and humid populations. A number of neutrality tests were applied to find evidence of positive selection in our candidate gene set, but only AcAQP2 gene in Pedregoso and San Ramón populations revealed significant departures from neutrality with positive values suggesting balancing selection. Conclusions: In this study we report the levels of nucleotide diversity searched in some drought stress candidate genes in Austrocedrus chilensis and the selective factors that may be acting on this species.

Effects and mechanisms of platycladi cacumen carbonisatum on rats with blood-heat and hemorrhage syndrome / 中国中药杂志

<p><b>OBJECTIVE</b>To discuss the effect and mechanism of Platycladi Cacumen Carbonisatum (PCC) on rats with blood heat and hemorrhage syndromes.</p><p><b>METHOD</b>Rats were fed with 15 g x kg(-1) water decoctions of Zingiberis Rhizoma and 5% alcohol for 15 days to establish the blood-heat and hemorrhage syndrome model. Yunnan Baiyao was taken as the positive control drug, and PCC decoctions (5.0, 10.0 g x kg(-1)) were given simultaneously, in order to detect changes in general physical signs of rats, such as body weight, daily diet, volume of daily drinking and urine and stool, and rectal temperature. Automatic hematology analyzers was used to determine white blood cell (WBC), red blood cell (RBC), hemoglobin (HGB), and hematocrit (HCT), blood time by docking (BT). Blood rheometers was used to detect whole blood and plasma viscosities, thrombin time (TT), activated partial thromboplastin time (APTT), prothrombin time (PT) and fibrinogen content (FIB). Indexes related to thyroid functions, such as triiodothyronine (T3), tetraiodothyronine (T4), reverse triiodothyronine (rT3) and thyroid stimulating hormone (TSH) were measured by radio-immunoassay, and changes in lung tissues were observed by hematoxylin-eosin (HE) stain.</p><p><b>RESULT</b>After modeling, rats witnessed slow-down in weight growth rate, significant increase in daily diet, volume of daily drinking, urine and temperature, significant decrease in stools and their water content (P < 0.05, P < 0.01), rise in plasma T4 level, notable growth in T3 and rT3 concentrations (P < 0.05), decline in TSH concentration. Additionally, their WBC, RBC, HGB and HCT remarkably increased (P < 0.05, P < 0.01), with significant increase in high, middle and low whole blood viscosities and plasma viscosity (P < 0.01); their BT, TT, APTT were notably prolonged (P < 0.01), with significant increase in FIB content (P < 0.01). After oral administration of Yunnan Baiyao or PCC, rats of all groups showed significant improvement in blood heat syndromes (P < 0.05, P < 0.01), and their blood coagulation indexes including BT, TT, APTT, FIB, thyroid function indexes including T4, T3, rT3, TSH, WBC, RBC, HGB, HCT, whole blood viscosity and plasma viscosity were getting normal (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>PCC can ameliorate blood heat symptoms and pathologic hemorrhage among rats with blood heat and hemorrhage syndromes by inhibiting thyroid functions and correcting hemorheological and coagulation disorders.</p>

Screening of some ethnobotanically important plants of Nagar area Gilgit-Baltistan, Pakistan

Certain species of ethnobotanical importance belonging to families Asteraceae, Berberidaceae, Cupressaceae, Elaeagnaceae, Gentianaceae, Salicaceae and Tamaricaceae, were collected from Gilgit during the months of June and July 2008 and were investigated for the presence of alkaloids, amino acids, anthraquinone [free and as glycosides] ascorbic acid, carbohydrates, coumarins, flavonoids, phenolics, proteins, saponins and steroids in their aqueous, ethanol and benzene extracts. Artemisia maritime L. [leaves] showed positive results for carbohydrates, coumarins, phenolics and proteins. Ephedra gerardiana Wall ex. Stapf. [stem] tested positive for alkaloids, ascorbic acid, coumarins, phenolics, proteins, saponins and steroids. Tamarix gallica L. tested positive for alkaloids, amino acids, anthraquinone as glycoside, ascorbic acid, carbohydrates, flavonoids, phenolics, proteins and steroids in stems, roots and leaves. Salix acmophylla Boiss. showed positive results for alkaloids, amino acids, anthraquinone [free and as glycosides] ascorbic acid, carbohydrates, flavonoids, phenolics, proteins, saponins and natural steroids. Hippophae rhamnoides L. showed positive results for alkaloids, amino acids, anthraquinone [free as glycosides] ascorbic acid, carbohydrates, coumarins, flavonoids, phenolics, proteins, saponins and steroids. Berberis glycocarpa Stapf. showed positive results for alkaloids, amino acids, ascorbic acid, carbohydrates, flavonoids, phenolics, proteins and steroids in stem, roots and leaves. Similarly Juniperus excelsa Wall ex. C.A. Meyer showed positive result for anthraquinone [both free and as glycosides], carbohydrates, phenolics, proteins, saponins and natural steroids

Analysis of flavonoids in platycladi cacumen by UPLC-MS / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the flavonoids in Platycladi Cacumen.</p><p><b>METHOD</b>The constituents in Platycladi Cacumen were determined by UPLC-MS. A Waters BEH C18 column (2.1 mm x 150 mm, 1.7 microm) was used with a gradient elution of methanol-water containing 0.2% formic acid. The mass spectrometer equipped with electrospay ionization source was used as defector and operated in data was collected under the negative ion modes.</p><p><b>RESULT</b>Eleven constituents were identified.</p><p><b>CONCLUSION</b>In this study, the main flavonoids in Platycladi Cacumen were separated by UPLC, and identified through the information of mass number and UV spectra. With the information of MS/MS from flavonoids, the modes of breaking up flavonoids were discussed. It is an accurate and effective method which can be applied for the constituent identification of Platycladi Cacumen.</p>

Optimization of supercritical CO2 extraction of plantaginis semen oil by response surface methodology and studies on its antioxidant activity / 中国中药杂志

<p><b>OBJECTIVE</b>The optimum conditions of SFE-CO2 extraction of Plantaginis Semen oil (SPO), the composition of SPO and its antioxidant activities of SPO were all investigated in this paper.</p><p><b>METHOD</b>Response surface method (RSM) was used to establish the mathematical model of SFE-CO2 extraction of SPO to obtain the optimum conditions based on Single factor experiments. Fatty acid compositions and contents of SPO were tested by GC-MS, and antioxidant activities of SPO were studied by DPPH and ABTS free radical elimination method.</p><p><b>RESULT</b>The optimum conditions obtained through RSM analysis were as follows: extraction tempreture 70 degrees C, extraction pressure 30 MPa, extraction time 120 min and flow rate 30 L x h(-1). Under the optimal condition, predicted value was 35.91%, while the experimental value was 35.07%. The experimental values agree with the predicted from the regression model with a relative error less than 5%. The main components of SPO were 9,12,15-octadecatrienoic acid, 8,11-octadecadienoic acid, octadecanoic acid and hexadecanoic acid. Most of the fatty acids were polyunsaturated fatty ones, whose quantities were obtained more than 88%. The IC50DPPH and IC50ABTS were 1.13, 3.57 g x L(-1) respectively in DPPH and ABTS assay.</p><p><b>CONCLUSION</b>Optimization of the extraction process by RSM of SPO is convenient and feasible. SPO has good antioxidant activity and is worth to develop for application.</p>

Baking method of Platycladi Cacumen Carbonisatum based on similarity of UPLC fingerprints / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a baking method of Platycladi Cacumen Carbonisatum for providing a new idea to Carbonic Herbs' research.</p><p><b>METHOD</b>Samples were prepared in an oven for different time at different temperatures separately. Then the fingerprints of the samples were determined by UPLC. According to the standard fingerprint, the similarities of the samples' fingerprints were compared.</p><p><b>RESULT</b>The similarities of 3 samples, which were baked at 230 degrees C for 20 min, 30 min and at 240 degrees C for 20 min, were above 0.96.</p><p><b>CONCLUSION</b>According to the similarities of the fingerprints and in view of the appearances, Platycladi Cacumen Carbonizing should be baked at 230 degrees C for 20 min.</p>

Antiviral activity of obtained extracts from different parts of Cupressus sempervirens against herpes simplex virus type 1

The aim of this study was to search for new antiviral agents from herbal medicines. Ethanol extracts of C. semipervirens, C. semipervirens var. horizontalis and C. semipervirens cv. Cereiformis were used in experiments to test their influence on herpes viruses [HSV-1]. HeLa cells monolayers were infected with herpes viruses [HSV-1]. Antiviral activity of the plant extracts assessed using Hematoxylin and Eosin method and observed under a light microscope. All tests were compared with a positive control, acyclovir. showed that all three plants have antiviral activity against HSV-1 virus. The most active extract was the obtained extract from C. semipervirens. Among the different parts of this medicinal plant tested, the fruit's extract appeared to possess the strongest anti- HSV activity. In, of the extracts tested in this survey all showed significant antiviral potency

Determination of total saponin in Semen Platycladi and defatted powder of Semen Platycladi / 中国中药杂志

<p><b>OBJECTIVE</b>To determine the contents of total saponin in the unprocessed Semen Platycladi and defatted powder of Semen Platycladi.</p><p><b>METHOD</b>Through contrasting different coloration, proper condition was selected by using orthogonal test design. Spectrophotometry was established to determine the contents of total saponin in the unprocessed Semen Platycladi and defatted powder of Semen Platycladi.</p><p><b>RESULT</b>Coloration system of vanillin-acetic acid-sulfuric acid was determined. The proper condition of coloration was that sulfuric acid was 80%, water bathed 85 and heated for 10 minutes. The contents of total saponin in unprocessed Semen Platycladi was 0.142% the content of total saponin in the defatted powder of Semen Platycladi was 0. 319%.</p><p><b>CONCLUSION</b>This method, stable, simple and quick, is applicable to the determination of total saponin in Semen Platycladi. A Little of total saponin is lost in the processing course of Semen Platycladi.</p>

Anatomical studies on cones of Cupressus sempervirens Linn.

The cones of Cupressus sempervirens Linn., belonging to the family Cupressaceae were collected from Govt. Botanical Garden, Ootacamund and dried under shade. The dried cones were powdered and used for the determination of physico-chemical constants. The macroscopical and microscopical characters were also evaluated by using fresh cones and reported. The anatomical parameters of the cones were studied for the first time from this plant

Studies on quality control of Xueyakang capsule / 中国中药杂志

<p><b>OBJECTIVE</b>To establish the quality control standard for Xueyakang capsule.</p><p><b>METHOD</b>Radix Rehmanniae, Cacumen Platycladi, Folium Artemisiae Argyi, Folium Nelumbinis were identified by TLC, and the content of quercitrin was determined by HPLC.</p><p><b>RESULT</b>The TLC sports developed was fairly clear, the HPLC method showed good repeatability, and the average recovery of quercitrin was 100.7% with RSD 2.0%.</p><p><b>CONCLUSION</b>The method is simple, accurate and can effectively control quality of Xueyakang capsule.</p>

Protective effects of total flavones of metasequosia on cerebral ischemia-reperfusion injury in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the protective effects of total flavones of metasequosia (TFM) on cerebral ischemia-reperfusion injury in rats.</p><p><b>METHOD</b>The bilateral vertebral arteries of rats were occluded under anesthesia, and 4-5 h later the carotid arteries of rats in the conscious condition were occluded for 30 min followed by reperfusion for 90 min. The effects of TFM on the contents of water, Na+, Ca2+ in cortex, malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in brain hemispheres, as well as the EEG activities were observed.</p><p><b>RESULT</b>After 30 min ischemia and 90 min reperfusion, the contents of water, Na+ , Ca2+ and MDA were increased, and the SOD activity was reduced with abnormal EEG activity and ischemic injury in the brain tissues. TFM 25-100 mg x kg(-1) ip 30 min before the carotid arteries were occluded, decreased the elevated water, Na+, Ca2+ and MDA contents, increased the SOD activity, reduced the ischemic injury of brain tissue, and promoted the recovery of EEG activities.</p><p><b>CONCLUSION</b>TFM has a protective effect on cerebral ischemia-reperfusion injury in rats.</p>

Aerobiological Study for Airborne Plooern and Mold in Kuri-shi, Kyunggi-Do / 소아알레르기및호흡기학회지

In a large number of a1lergic cases, inhalant allergens, particularly airborne pollens and mo1ds am important causative, triggering agents in respiratory, even in skinanergy. It is essential for diagnosis and treatment of airborne allergy to survey the airborne pollen and mo1d in the patient's living area. Rotorod samp1er was installed on the well-ventilated rooftop of Hanyang Kuri Hospital in Kuri-shi, Kyunggi-do as the vicinity of Seoul. Airborne allergens were collected from October 1, 1995 to September 30, 1996 daily and were stained with Calberla`s fuchsin and were identified, counted under the light microscope (400Xfor pollen, 1000Xfor mold). The data were compared with the local daily weather report. The airborne pollen was found from 12th February to 22nd December. The maximal period for pollen collection was 7th-25th May (Maximal count:492 grains/ August. There were Alnus, Betula, Pinus, Quercus, Acer, Ulmus, PopuluS, Cupressaceae, Ginkgo, Corylus, Salk in the tree season from middle of February to middle of July. The grass season ranged from middle of March to end of November. There were Artemisia, Ambrosia, HumuluS, Chenopodium-Amaranthus, rumex, Plantago, Trifoium, Humulus, Chrysanthemum, Cyperus in the weed season ranged from middle of July to end of December. Airborne molds were collected all through the year except JanuaIgamma but, counted maximally from the middle of June to end l August. There were CladosPorium, Alternaria, ascospore of Leptospheria, Periconia, Myxomycete, Drechs1era, Rust urediniospores, Basidiospore, ToruIa, Xylariaceae, IOidimn, Nigrospom, Stempi1y1ium, Amphispheda, Fusarium, Aspergil1us. In Korea, there are great number of the allergic patients, but we have had a little records and data about study of relationship between the allergic diseases and airborne environmental allergens. we will reported those results completely in future.