Evaluation and application of an efficient plant DNA extraction protocol for laboratory and field testing / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Nucleic acids in plant tissue lysates can be captured quickly by a cellulose filter paper and prepared for amplification after a quick purification. In this study, a published filter paper strip method was modified by sticking the filter paper on a polyvinyl chloride resin (PVC) sheet. This modified method is named EZ-D, for EASY DNA extraction. Compared with the original cetyl trimethylammonium bromide (CTAB) method, DNA extracted by EZ-D is more efficient in polymerase chain reaction (PCR) amplification due to the more stable performance of the EZ-D stick. The EZ-D method is also faster, easier, and cheaper. PCR analyses showed that DNA extracted from several types of plant tissues by EZ-D was appropriate for specific identification of biological samples. A regular PCR reaction can detect the EZ-D-extracted DNA template at concentration as low as 0.1 ng/μL. Evaluation of the EZ-D showed that DNA extracts could be successfully amplified by PCR reaction for DNA fragments up to 3000 bp in length and up to 80% in GC content. EZ-D was successfully used for DNA extraction from a variety of plant species and plant tissues. Moreover, when EZ-D was combined with the loop-mediated isothermal amplification (LAMP) method, DNA identification of biological samples could be achieved without the need for specialized equipment. As an optimized DNA purification method, EZ-D shows great advantages in application and can be used widely in laboratories where equipment is limited and rapid results are required.

Color characterization and its correlation with quality index during ripening of Rubus chingii / 中国中药杂志

The color of Rubus chingii was characterized by digital method, and the content of water extract, alcohol extract, total flavonoids, total polysaccharides, total polyphenols, ellagic acid, linden glycoside, kaophenol-3-O-rutin were determined. Correlation regression was used to analyze the correlation between color and composition. The results showed that L~* was positively correlated with total polyphenols, kaophenol-3-O-rutin and tilide, and moderately positively correlated with total flavones, ellagic acid and aqueous extracts. The a~* value was negatively correlated with total polyphenols, kaophenol-3-O-rutin, and linden glycosides, while was moderately correlated with total flavones, aqueous extracts, and ellagic acid. The b~* value was negatively correlated with the water extract, and moderately correlated with the content of total polyphenols, total polysaccharides, alcohol extract and kaophenol-3-O-rutin, which showed that R. chingii mature color had a significant correlation with material composition in the process of dynamic change. According to the law of dynamic change in the color and quality indexes, it is determined that the appropriate harvest time is in late April to May 1, while the fruit is not turn yellow. The agronomic traits related to fruit was(12.49±0.56) mm in diameter,(14.25±1.19)mm in height,(1.20±0.14) g in weight, the chroma L~* value was 52.87±3.14,a~* value was 2.01±1.58, b~* values was 28.31±3.88. The results lay a foundation for establishing an objective quantitative evaluation model of R. chingii color from experience.

Breeding and variety characteristics of a new variety of Fritillaria thunbergii "Zhebei 3" / 中国中药杂志

A new variety "Zhebei 3(Zhejiao Pharmaceutical 2018002)" was selected and bred from multi seeded Fritillaria thunbergii mutants by systematic breeding method. From 2012 to 2016, the traits assessment, disease resistance appraisal, plot ratios and regional trials of the variety were continuously carried out. The results showed that "Zhebei 3" emerged early and had late seedlings. The average growth period was about 100 days, which was 6 days and 12 days higher than the "Zhebei 1" and "Zhebei 2". The average yield was 5 095.5 kg·hm~(-2), which was 14.42% and 17.71% higher than of the control respectively. The average proliferation rate of bulbs was 261.2%, which was 37.46% and 31.58% higher than that of the control, respectively. The propagation coefficient of bulbs was about 1∶2.6, and the total amount of peimine and peiminine was 0.172 2%, which was 4.49% and 29.47% higher than the control, respectively. The identification of disease resistance showed that it was resistance to bulb stem(soft) rot, better than the control. "Zhebei 3" has stable characters, high yield, good quality, strong disease resistance, and moderate propagation coefficient which is suitable for planting in Zhejiang province.

Correlation analysis between quality and habitat phenotype of Hovenia acerba / 中国中药杂志

The contents of dihydromyricetin and total flavonoids of Hovenia acerba seeds were detected by HPLC-DAD and UV spectrophotometer. And then the correlation between the habitat, phenotype and quality of H. acerba seeds were deeply studied. There were big differences in both appearance and quality among the H. acerba seeds from different places. It showed that the content of dihydromyricetin in H. acerba seeds was 0.41-9.81 mg•g⁻¹, and the content of total flavonoids was 5.52-21.98 mg•g⁻¹. The cluster analysis showed that the quality of H. acerba seeds was related to the habitat. The samples from Jianghan Plain Area showed relatively stable and excellent quality. According to the correlation analysis, there was a significant positive correlation between 1 000-seeds weight, red-black seeds ratio and the content of dihydromyricetin, while the content of total flavonoids was not related to the 1 000-seeds weight and red-black seeds ratio. So the 1 000-seeds weight and red-black seeds ratio could be used as an initial judgment of the quality of H. acerba seeds. As dihydromyricetin and total flavonoids content was not related, both dihydromyricetin and total flavonoids should be taken into account when the quality of H. acerba seeds was studied.

Study on combined effects of chemical components for different flowers blossoming degree of yellow medicinal Chrysanthemum morifolium from Zhejiang / 中国中药杂志

The ingredients of five kinds of Zhejiang's yellow Chrysanthemum morifolium with different flower blossoming stages were comparatively analyzed. Polysaccharides, total flavonoids, volatile oil, alcohol extract, water extract, chlorogenic acid, luteolin, 3,5-O-dicaffeoyl quinic acid and fingerprint of the ingredient were determined as indicators. During flower blossoming stages, the ingredients of Ch. morifolium showed a big difference with a certain variation. At the early opening stage, the contents of flavonoids and volatile oil were higher, the content of chlorogenic acid, luteolin, 3,5-O-dicaffeoyl quinic acid were higher in the middle of the flowers 50% -80% fowers blossoming degree is the optimal time for harvest.

Study on method of paeonia radix alba producing and concocting integration processing / 中国中药杂志

<p><b>OBJECTIVE</b>To study the origin pre-treating and processing integration techniques of Paeonia Radix Alba.</p><p><b>METHOD</b>Different processing integration techniques were adopted and compared with traditional processing techniques to determine drying rate, aqueous extracts and peoniflori content.</p><p><b>RESULT</b>Half-dry slices baked at 100 degrees C for 20 min and steamed at 100 degrees C for 10 min had the highest peoniflori contents. Half-dry slices baked at 100 degrees C for 20 min had the highest content of aqueous extracts. Products processed with conventional method and sulfur-fumigation had the lowest content of aqueous extracts.</p><p><b>CONCLUSION</b>The origin processing integration techniques of Paeonia Radix Alba lose less active ingredients than conventional processing methods.</p>

Study on method of Corydalis yanhusuo producing and concocting integration processing / 中国中药杂志

<p><b>OBJECTIVE</b>To establish integration processing method for pretreating and vinegar producing Corydalis yanhusuo.</p><p><b>METHOD</b>Different processing methods were contrasted with the traditional processing technology, and contents of corydalis B, water extract and ethanol extract in samples of different processing products were determined.</p><p><b>RESULT</b>The content of corydalis B were best in the samples of vacuumizing C. yanhusuo chips scaked in rice vinegar for twice or soaked in rice vinegar after chip drying. The water extract was highest in the samples of chip soaked in rice vinegar after drying, followed with chip vacuumizing twice, and there were no remarkable difference between the other samples and the traditional process. The difference of ethanol extract was not remarkable in all the samples.</p><p><b>CONCLUSION</b>The study provide the feasibility of C. yanhusuo producing and concocting integration processing.</p>

Effect of environmental factors on growth of Huperzia serrata / 中国中药杂志

<p><b>OBJECTIVE</b>To establish and optimize the cultivation systems for wild Huperzia serrata, through assessing the effects of environmental factors on its growth.</p><p><b>METHOD</b>H. serrata was planted under natural field condition, the growth status of H. serrata under different light intensities, temperatures, relative humidity (R. H.), fertilizer levels, and so on were investigated and analyzed.</p><p><b>RESULT</b>H. serrata grew nicely under following environmental conditions: temperature ranging from 18.9-26.3 degrees C, R. H. ranging from 81%-90% and 1330-3000 lx light intensity. A cultivation system, in which H. serrata was planted in the bamboo forest and did not occupy the arable land, was established through the study.</p><p><b>CONCLUSION</b>Concluded that light intensity and R. H are the two factors which greatly confine the growth of H. serrata. As long as the levels of the above two factors meets the demands, H. serrata could be cultivated.</p>

Study on correlation between Atractylodes macrocephala viability and germination rate / 中国中药杂志

<p><b>OBJECTIVE</b>To screen the optimized methods for detection seed viability and germination rate determination of Atractylodes macrocephala, and determine the relationship between seed viability and germination rate.</p><p><b>METHOD</b>There were four methods, which including 2,3,5-triphynel tetrazolilum chloride (TTC) staining, red ink staining, BTB staining and Nongjia method, to evaluate the 12 A. macrocephala local varieties'seed viability and measure their germination rate.</p><p><b>RESULT</b>Seed viability of A. macrocephala using TTC staining ranked the first compared to that of other three methods. Seed viability was significantly related with germination rate using TTC method. Their correlation coefficient reached 0.915 and regression equation was also found out between seed viability (X) and germination rate (Y), which was Y = -0.083 4 + 0.995 4X.</p><p><b>CONCLUSION</b>TTC staining was the optimal method to determine A. macrocephala seed vitality. Furthermore, seed viability was significant related with germination rate of A. macrocephala.</p>

Mutagenic effects of gamma-rays on Coix lacryma-jobi var. ma-yuen / 中国中药杂志

<p><b>OBJECTIVE</b>To study the mutagenic effect of gamma-rays on Coix lacryma-jobi var. ma-yuen.</p><p><b>METHOD</b>Physiological and mutagenic effects of gamma-rays on C. lacryma-jobi var. ma-yuen dormant seeds were studied. The germination percentage, seeding survival, seeding height and root length of M1 plants and the frequency of chlorophyll mutation in M2 generation were selected as criteria.</p><p><b>RESULT</b>The gamma-rays showed obvious inhibitory action to the seedling growth, and a strong ability in inducing the chlorophyll mutation.</p><p><b>CONCLUSION</b>The gamma-rays is one kind of C. lacryma-jobi var. ma-yuen effective mutagen. The appropriate dose of gamma-rays is 450 Gy for C. lacryma-jobi var. ma-yuen dormant seeds.</p>

GC-MS analysis of essential oil from anthodiums of Chrysanthemum morifolium processed by microwave-airflow and steam calefaction / 中国中药杂志

<p><b>OBJECTIVE</b>Analysis of the constituents of the essential oil extracted from the flowers of the Chrysanthemum morifolium processed by the microwave-airflow and the calefaction after steam process from the State Chrysanthemum GAP bases in Zhejiang Province and to provide scientific basis for quality control.</p><p><b>METHOD</b>The essential oil was extracted by water-steam distillation and separated by GC capillary column chromatography. The components were quantitatively determined with normalization method, and identified by GC-MS.</p><p><b>RESULT</b>From the microwave-airflow dried sample and the calefactively dried sample after steam process, 119 and 175 components were detected, among them fifty and fifty-five components were identified, which were composed of 67.89% and 63.64% of the total essential oil, respectively.</p><p><b>CONCLUSION</b>The yield of the essential oil extracted from the microwave-airflow dried sample was 0.40%, and that from the calefactively dried sample after steam process was 0. 19%. The former is 2.1 times higher than the latter. The components of the essential oil are similar and there are differences between the content of corresponding compounds in the two samples. The results showed that application of microwave-airflow combined drying technology remained original and essential constituents. The production benefit was improved greatly. The grade and quality of C. morifolium enhanced, and it's market selling price was increased compared to the traditional drying calefaction after steam process.</p>

Study on growth characteristics of Curcuma wenyujin / 中国中药杂志

<p><b>OBJECTIVE</b>To study on growth characteristics of Curcuma wenyujin, and provide theoretical basis for the development of high-quality and high-yield medical material.</p><p><b>METHOD</b>The morphological change of the plant was observed periodically, the content of volatile oil and dry matter in leaves, rhizome, root tuber was determine.</p><p><b>RESULT</b>The growth of C. wenyujin could be divided into 5 stages, i. e. seed germination, seedling, leaf growth, rhizome expansion, accumulation of dry matter, respectively. Before the stage of rhizome expansion, over 70% dry matter was accumulated in the aerial part of the plant, and during the stage of leaf growth, the maximum increase rate of dry matter in aerial part was 3.90 g/p/d. During the stage of rhizome expansion, the ratio of dry matter of rhizome increased quickly and reached above 33% , and the increase rate of dry matter of rhizome rise up to 3.83 g/p/d, in the end of the stage, the content of volatile oil in the rhizome also rose up to 1.20 mL x 100 g(-1).</p><p><b>CONCLUSION</b>During the whole growth stage, there are two growth centers, when the two curves of dry matter increase of aerial part and rhizome intersect, it is regarded as a signal that the growth transformed form the aerial part to rhizome. When the rate of dry matter from rhizome rise, the content of volatile oil in rhizome rises quickly with the increase of dry matter in rhizome. The optimal harvest time is in mid-December.</p>

Effects of agmatine on the electrical activity of subfornical organ neurons / 生理学报

The aim of this study was to investigate the effects of agmatine (Agm) on the electrical activity of neurons in subfornical organ (SFO) slices using extracellular recording technique. The results are as follows. (1) In response to the application of Agm (1.0 micromol/L) into the superfusate for 2 min, the discharge rate of 24/28 (85.7%) subfornical neurons was decreased significantly, while the discharge rate of 4/28 (14.3%) neurons were not affected. (2) Pretreatment with L-glutamate (0.3 mmol/L) led to a marked increase in the discharge rate of 19/24 (79.2%) subfornical neurons in an epileptiform pattern and the activity of the remaining 5/24 (20.8%) neurons was unaffected. By application of Agm (1.0 micromol/L) into the superfusate for 2 min, the epileptiform dicharge of 15/19 (78.9%) neurons was suppressed significantly, while that of the other 4 (21.1%) neurons was not inhibited. (3) In 12 neurons, perfusion of the selective L-type calcium channel agonist, Bay K-8644 (0.1 micromol/L), induced a significant increase in the discharge rate of 10/12 (83.3%) neurons, while the other 2 (16.7%) neurons showed no change. The increased discharge of 8/10 (80%) neurons was reduced by application of Agm (1.0 micromol/L) into the superfusate and that of 2/10 (20%) neurons was not affected. (4) Application of nitric oxide synthase (NOS) inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME, 50 micromol/L) into the superfusate also significantly increased the discharge rate of 6/9 (66.7%) neurons, and that of 3/9 (33.3%) neurons had no response. Agm (1.0 micromol/L) applied into the superfusate reduced the increased discharge of all 6/6 (100%) neurons. These results suggest that Agm can inhibit the spontaneous discharge, and L-glutamate, Bay K-8644- or L-NAME-induced discharge of neurons in SFO. These inhibitory effects of Agm may be related to the blockade of NMDA receptors and reduction in calcium influx in SFO neurons.

Effects of agmatine on neuronal discharges in rat hippocampal CA1 area / 生理学报

The effects of agmatine (Agm) on the discharges of neurons in CA1 area of hippocampal slices were examined by using extracellular recording technique. The results are as follows. (1) In response to the application of Agm (0.1-1.0 micromol/L) into the superfusate for 2 min, the spontaneous discharge rates (SDR) of 38/47 (80.9%) neurons were decreased significantly in a dose-dependent manner, while that of 9/47 (19.1%) neurons showed no change in discharge rate; (2) pretreatment with L-glutamate (L-Glu, 0.2 mmol/L) led to a marked increase in SDR of 9/12 (75%) neurons in an epileptiform pattern and that of 2/12 (25%) neurons were not affected, then after Agm (1.0 micromol/L) was applied into the superfusate for 2 min, the epileptiform discharges were suppressed significantly; (3) in 7 neurons, perfusion of the selective L-type calcium channel agonist, Bay K-8644 (0.1 micromol/L), induced an increase in the SDR of 6/7 (85.7%) neurons, while that of 1/7 (14.3%) neuron showed no change, and the discharges were also decreased by application of Agm (1.0 micromol/L) into the superfusate; and (4) application of NOS inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME, 50 micromol/L) into the superfusate 5 min later also significantly increased the SDR in all 13 (100%) neurons; then Agm (1.0 micromol/L) applied into the superfusate inhibited the discharges of 11/13 (84.6%) neurons, while those of 2/13 (15.4%) neurons were not affected. These results suggest that agmatine can inhibit the spontaneous discharges and L-glutamate-, Bay K-8644- and L-NAME-induced discharges of hippocampal CA1 neurons. These inhibitory effects of agmatine may be related to the blockade of NMDA receptors and a reduction in calcium influx in hippocampal neurons

Determinatin of Stachydrine Hydrochloride in Fresh Medicinal Material of Herba Leonuri by HPLC / 中药新药与临床药理

Objective To establish a HPLC method for the separation and determination of stachydrine hydrochloride in fresh medicinal material of Herba Leonuri.Methods The NH2 column(250 mm? 4.6 mm,5 ? m) was adopted with acetonitrile-water(80 ∶ 20) as mobile phase at a flow rate of 1.0 mL/min.The detection wavelength was at 192 nm.The column temperature was at 22 ℃.The solution for content determination was obtained after extracted with ethanol and then filtered and purified through an activated charcoal-aluminium oxide column with ethanol as eluant.Results Stachydrine hydrochloride in fresh Herba Leonuri can be separated and determined on NH2 column.A good linear range was within 2.29～ 11.46 ? g.The average recovery for stachydrine hydrochloride was 98.26 % and RSD was 1.52 %(n=5).Conclusion The method is steady and with good repeatability,and can be used to determine the content of stachydrine hydrochloride in fresh Herba Leonuri.