A Comparative Transcriptomic with UPLC-Q-Exactive MS Reveals Differences in Gene Expression and Components of Iridoid Biosynthesis in Various Parts of Gentiana macrophylla.

Gentiana macrophylla Pall. (G. macrophylla)-a member of the family Gentianaceae-is a well-known traditional Chinese medical herb. Iridoids are the main active components of G. macrophylla, which has a wide range of pharmacological activities such as dispelling wind, eliminating dampness, clearing heat and asthenic fever, hepatoprotective and choleretic actions, and other medicinal effects. In this study, a total of 67,048 unigenes were obtained by transcriptomic sequencing analysis of G. macrophylla. A BLAST analysis showed that 48.21%, 33.66%, 46.32%, and 32.62% of unigenes were identified in the NR, Swiss-Prot, eggNOG, and KEGG databases, respectively. Twenty-five key enzymes were identified in the iridoid biosynthesis pathway. Most of the upregulated unigenes were enriched in flowers and leaves. The trustworthiness of the transcriptomic data was validated by real-time quantitative PCR (qRT-PCR). A total of 22 chemical constituents were identified by ultra-high performance liquid chromatography-quadrupole-electrostatic field Orbitrap mass spectrometry (UPLC-Q-Exactive MS), including 10 iridoids. A correlation analysis showed that the expression of 7-DLH and SLS was closely related to iridoids. The expression of 7-DLH and SLS was higher in flowers, indicating that flowers are important for iridoid biosynthesis in G. macrophylla.

Association between the ABCA1 (R219K) polymorphism and lipid profiles: a meta-analysis.

Conflicting evidence was found about the relationship between lipid profiles and R219K polymorphism in adenosine triphosphate-binding cassette exporter A1 (ABCA1) gene. In this study, four meta-analyses were conducted to assess the effect of R219K on lipid levels, including high-density lipoprotein cholesterol (HDLC), low-density lipoprotein cholesterol, total cholesterol, and triglycerides (TG). A total of 125 samples of 87 studies (about 60,262 subjects) were included. The effect of each study was expressed using the standard mean difference (SMD) and 95% confidence interval (95% CI) and pooled by meta-analysis in the random-effects model. Subgroup and meta-regression analyses were conducted to explore potential heterogeneity sources. The overall pooled effect showed the following results. (1) The R219K was significantly associated with HDLC level (SMD = - 0.25 mmol/L, 95%CI - 0.32 to - 0.18, z = - 6.96, P < 0.01, recessive genetic model). People with different genotypes had significantly different HDLC levels under the recessive, codominant and dominant genetic models (all Ps < 0.01). (2) A weak and indeterminate relationship between R219K and TG level was observed (SMD = 0.18 mmol/L, 95%CI 0.06-0.30, z = 3.01, P < 0.01, recessive genetic model). These findings suggested that R219K was associated with HDLC and TG levels, which might implicate a promising clinical application for lipid-related disorders, though the influences of race, health status, BMI, and other heterogeneity sources should be considered when interpreting current findings. The protocol was registered at PROSPERO (registration number: CRD42021231178).

Study on effects of sweating treatment and different drying methods on the quality of Gentianae macrophyllae Radix.

To provide basis for improving the quality of Gentianae macrophyllae Radix and optimising the field processing method. Gentianae macrophyllae Radix samples were collected from Long County. Five main active iridoids were determined by using HPLC. The HPLC fingerprints were measured and analysed by the traditional Chinese medicine chromatographic fingerprint similarity evaluation system. Water, total ash, acid insoluble ash and ethanol extract contents were determined in accordance with Chinese Pharmacopoeia (2015). Colour of the surface and powdered samples were determined by using a colorimeter. Data were analysed by SPSS11.0. Notable differences were found among samples from different sweating and drying processes, in spite of the relatively consistent overall chemical characteristics. Sweating and drying methods influenced the contents of some active ingredients and colour very significantly; moreover, they also showed significant effects on the water, ash, and ethanol extract contents. The necessity of sweating treatment in the field processing of Gentianae macrophyllae Radix may be insufficient. Drying in the shade and oven drying are more profitable for preserving the active constituents. Gentianae macrophyllae Radix should be dried directly without sweating, and drying in the shade and oven drying should be adopted preferentially to improve the quality.

[Comparison research on quality of cultivated and wild Anemarrhena Rhizome from Yi County].

OBJECTIVE: To compare the quality of cultivated and wild Anemarrhena Rhizome from Yi County (Xiling Zhimu) based on contents analysis of active constituents. METHOD: Samples of cultivated Anemarrhena Rhizome from most townships of Yi County were analyzed and compared with wild ones. Six indexes belonged to three kinds active constituents of saponin, flavornoid and polysaccharide were adopted. HPLC-ELSD method with cholesterol as internal standard was adopted to determine the content of sarsasapongenin. HPLC-ELSD method was used to simultaneously determine the contents of anemasaponin C and Anemasaponin A III. Contents of neomangiferin and mangiferin were determined by HPLC-UV method. Total polysaccharide was determined by phenol sulfate method. RESULT: The mean content of sarsasapongenin in cultivated Anemarrhena Rhizome samples is slightly lower than the wild. The mean contents of anemasaponin C and Anemasaponin A III in cultivated Anemarrhena Rhizome samples are higher than the wild. There is no notable difference of these three index between the cultivated and the wild. The cultivated Anemarrhena Rhizome samples have a lower content of neomangiferin and a higher content of mangiferin than the wild. While the total content of these two flavonoids have no notable difference. The cultivated Anemarrhena Rhizome samples have a higher content of total polysaccharide than the wild samples. CONCLUSION: Contents of active constituents in cultivated Anemarrhena Rhizome from Yi County (Xiling Zhimu) are not notably different with the wild Anemarrhena Rhizome. They have similar good quality as the wild ones.

[Simultaneous determination of two saponnins in anemarrhenae rhizoma by HPLC-ELSD].

OBJECTIVE: To establish an HPLC-ELSD method for determination of Anemarsaponin C and Anemarsaponin A III in Anemarrhenae Rhizoma. METHOD: Kromasil C18 column(4.6 mm x 250 mm, 5 microm) was used as stationary phase. Mobile phase was methanol-water gradient with the flow rate of 1 mL x min(-1); the temperature of the drift tube and evaporation was 50 degrees C and 70 degrees C respectively. The gas pressure was 1.03 x 10(5) Pa. RESULT: There are good linearity in the range 0.310-3.10 microg of anemarsaponin C (lgA = 1.254 2lgM + 5.734 7, r = 0.999 5) and in the range 0.323-3.23 microg (lgA = 1.328 41gM + 5. 937, r = 0.999 6) of anemarsaponin A III. The average recovery of anemarsaponin C and anemarsaponin A III was 98.1% with RSD 2.1% and 97.3% with RSD 1.5% (n = 6) respectively. CONCLUSION: The method is rapid and accurate. It is suitable for quality control of Anemarrhenae Rhizoma. The result of determination reveals that the quality of Anemarrhenae Rhizoma from different places of north China are of notable difference.

[Study on the chemical constituents in roots of Gentiana dahurica].

OBJECTIVE: To systematically study the chemical constituents in the roots of Gentiana dahurica. METHODS: Various column chromatographic techniques were used for isolation and purification. The structures were elucidated on the basis of spectral data (UV, IR, MS, NMR) and identified by comparing with the authentic substance. RESULTS: Seven compounds were isolated and identified as: roburic acid (1), oleanolic acid (2), beta-sitosterol (3), daucosterol (4), gentiopicroside(5), swertiamarine (6), sweroside (7). CONCLUSION: Compounds 1, 2 and 4 are isolated from this plant for the first time.

Separation and purification of triterpene saponins from roots of Radix phytolaccae by high-speed countercurrent chromatography coupled with evaporative light scattering detection.

Coupled with evaporative light scattering detection, high-speed countercurrent chromatography was successfully applied for the first time to separation and purification of four triterpene saponins including esculentoside A, B, C and D from roots of Radix Phytolaccae. The separation was performed with an optimized two-phase solvent system composed of chloroform-methanol-water (4:4:2, v/v) using the lower phase as the mobile phase at a flow rate of 1.5 ml/min,. From 150 mg of crude extract 46.3 mg of esculentoside A, 21.8 mg of esculentoside B, 7.3 mg of esculentoside C, and 13.6 mg of esculentoside D were obtained at purities of 96.7%, 99.2%, 96.5% and 97.8%, respectively, as determined by HPLC analysis. The structures of the four triterpene saponins were identified by ESI-MS,(1)H NMR and (13)C NMR.

[Realization of algorithm on finishing optimization-tool-path generation for high-speed machining molar crown].

Molar crown is very small and has not only thin-wall, but also complex profile, especially, the occlusal surface of each molar crown has many cusps, ridges and fossae being differently distributed. When conventional processing method is used, it is impossible to machine molar prosthesis rapidly and exactly. To enhance machining velocity and improve the surface precision of molar crown, an algorithm of entity rapid offset-based STL format is put forward. By the application of Zigzag toolpath planning and micro-machining cutter, the finishing toolpaths for high speed milling molar prosthesis are generated. In terms of Mikron UCP800 high-speed machine center, the molar all-crown made of alloy aluminum material is successfully machined. The test results show that the algorithm of tool-path generation works fast, the number of toolpaths is small, and the cutter feeds smoothly.

Changes of gentiopicroside synthesis during somatic embryogenesis in Gentiana macrophylla.

IN VITRO plant regeneration of Gentiana macrophylla Pall. and determination of gentiopicroside content during somatic embryogenesis are described in the present work. The highest percentage of embryogenic callus formation was observed in Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg/L 6-benzylaminopurine (BA). Calli were subcultured on MS medium containing 1.0 mg/L 2,4-D, 1.0 mg/L BA and 500 mg/L lactalbumin hydrolysate (LH) at intervals of 25 days. A higher frequency of somatic embryo maturation was achieved on MS medium containing B5 vitamins (MB) supplemented with different concentrations of 1-naphthaleneacetic acid (NAA) and BA than with a combination of NAA and kinetin (KT). Addition of AgNO(3) improved maturation of somatic embryos while thidiazuron (TDZ) promoted vitrification. The gentiopicroside contents of embryogenic calli and globular-, heart-, torpedo-, and cotyledon-shaped embryoids were determined by high-performance liquid chromatography (HPLC). Gentiopicroside was not detectable in embryogenic calli, but in all types of somatic embryos. The highest gentiopicroside content was observed in cotyledon-shaped embryoids, reaching more than 12 mg/g dry weight.

[Research of tool-path generation algorithm for NC machining dental crown restoration].

Seeing that the manual method to restore tooth has the disadvantages such as long "lead-time", assurance of quality highly depending on operator's technology, and real-time cure difficulty met by lots of dental patients coming up for tooth restoration, we put forward an algorithm of tool-path generation based on STL data model for roughing dental restoration. The algorithm can reconfigure the STL data of dental crown restoration quickly, can generates the multi-level offset wire-loop by the use of horizontal plane cutting triangle facets; and then on the basis of offset wire-loop, it can plan Zigzag and follow the contour machining tool path. The algorithm has been applied to Dental CAM software, through simulation machining, the result shows that it can not only generate interference-free tool path, but also save a lot of "lead-time" for dental restoration. Accordingly, the algorithm is of great value for reference in clinical application.

[Studies on the chemical constituents in root of Gentiana macrophylla from Shaanxi].

OBJECTIVE: To in vestigate the constituents in root of Gentiana macrophylla. METHOD: Various column chromatographic techniques were used for isolation and purification of the principles. The structures were elucidated on the basis of spectral data (UV, IR, MS, 1H-, 13C-NMR etc.) and identified by comparing with standard substance. RESULT: Eight compounds were identified. Four compounds isolated from the chloroform fraction are: 5-carboxyl-3,4-dihydrogen-1H-2-benzopyran-1-one (1), erythrocentauric acid (2), roburic acid (3), oleanolic acid (4). Water fraction gave four known secoiridoid glucosides. They were: gentiopicroside (5), swertiamarine (6), sweroside (7), 6'-O-beta-D-glucosylgentiopicroside (8). CONCLUSION: 1 is a novel compound. It was named as erythrocentauric acid. 2 was isolated from genus Gentiana and 8 was isolated from G. macrophylla for the first time.

[Analysis of volatile constituents from Anemarrhena asphodeloides by GC-MS].

OBJECTIVE: To study the chemical constituents of the volatile oil from the rhizome of Anemarrhena asphodeloides. METHOD: The volatile oil was steam distillation. Chemical constituents were separated and analyzed by GC-MS. The relative content of each component was determined by area nomalization. RESULT: 24 volatile compounds were isolated and identified for the first time, representing 70.83% of the total oil. CONCLUSION: The main constituents of this oil were aldehydes (31.15%), terpene and their oxide (20.66%), alkyls (8.35%), Furan heterocyclic compounds (6.41%), non terpene alcohol (4.26%). There are 12 compounds with contents over 3%. Among them, borneol has the highest content (9.35%).