Macrophorins H and L, two new HMG-conjugate macrophorins from rihzospheric Penicillium sp. NX-05-G-3.

Macrophorins H (4) and L (5), two rare HMG-conjugate macrophorins along with three known macrophorins (1-3), three DMOA-derived meroterpenoids (6-8) and two ergosterol derivates (9-10) were isolated from sterilized rice medium cultured Penicillium sp. NX-05-G-3. Their structures were elucidated by 1D and 2D NMR. The cytotoxicities of all compounds were evaluated, and compounds 1 and 2 showed extensive cytotoxicity against human cancer cell lines Hela, SCC15, MDA-MB-453 and A549, with IC50 values ranging from 17.6 to 32.8 µM.

[Investigation and application of Dendrobium resources in Qinba Mountains with a discussion on industrial poverty alleviation model].

The Dendrobium species are rare and endangered medicinal plants, and it is difficult to investigate their wild resources with conventional methods because of typical epiphytic herbaceous. We explored Dendrobium resources(include culture resource) of Qinba Mountains and the boundary Mountain area in Hubei, Chongqing using the methods of literatures and field investigation, and found that the cultural base of Dendrobium were profound in Qinba Mountains region. Furthermore, its germplasm resources of Dendrobium were established for the first time in Wanzhou Luotian town. In case the advantages of local rock resources and poverty alleviation demand, we have actively carried out the cultivating mode of Dendrobium which grow on rock, and the poverty alleviation model of local characteristic Dendrobium industry were established preliminarily. Our application case can provide reference for the mining and transformation of traditional Chinese medicine resources census results.

[A developmental research on wild Dipsacus asper in Chongqing Wulong district].

In order to study the distribution and dynamics growth of wild Dipsacus asper resources in the Wulong district of Chongqing, 9 sample plots were selected for 12 consecutive months in the natural distribution area of the D. asper in Wulong district by using the sample line + plot survey method to conduct a field survey. The results showed that D.asper was distributed in forest edge wasteland or shrub-grassland, and growbetter with loose yellow-brownsoil or red soil, and poor with lithologic soil or impounded surface water.The growth curve of the plant height from June to July and the ground fresh weight from July to August showed a turning point, it might consume large amounts of nutrients during its flowering period, resulting in the restriction of vegetative growth.The highest temperature in the distribution area of D.asperoides in Wulong district is less than 30 °C, the minimum temperature is about 0 °C, and the rainfall is 1 241-1 392 mm. Its growth environment is no severecold in winter, no heat in summer, and abundant rainfall.The main growth stage of D.asper is from July to October, and the range of root dry rate was 0.162 5-0.239 7 in Xiangkou, 0.154 9-0.223 6 in Baima Mountain, and 0.143 7-0.203 3 Xiannv Mountain. The vegetative growth and dry matter accumulation synchronized in the main growth stage, and the accumulation rate of dry matter was faster than that of vegetative growth. The correlation analysis between indicators and root fresh weight showed that the fresh weight of the aerial part and root fresh weight had the best correlation.

[Analysis on ecological factors and active components content of wild Dipsacus asper in Chongqing Wulong district].

An HPLC method was developed for the determination of iridoid glycosides (loganin acid, loganin, sweroside) and saponins (asperosaponin Ⅵ) in the wild Dipsacus asper. A total of 108 samples consecutive growing 12 month were collected in 9 plots in Wulong district of Chongqing. Subsequent analysis of the content of loganin acid, loganin, sweroside and asperosaponin Ⅵ was performed by HPLC to evaluate the quality. In addition, 20 climate data provided by the world climate database (http://www.worldclim.org/) was analyzed to deduce the correlation between the growing environment factors and the active ingredient content accumulation of D. asperoides and choose the apposite growing environment for D. asper. The range of active ingredient content in wild D. asper were 0.01%-3.80%(loganin acid), 0.08%-0.62%(loganin), 0.12%-0.78%(sweroside), 0.64%-5.26%(asperosaponin Ⅵ). The highest content of these active ingredients was concentrated from February to April, with 2.64% of loganin acid, 0.36% of loganin), 0.57% of sweroside, and 3.09% of asperosaponin Ⅵ. The method used for determination of the active ingredient content in D. asper was simple and convenient with accurate result. The selection of the quadrats is scientific and reasonable and can be used for the analysis of the contents of the wild D. asper, thus provide a reference for quality evaluation of D. asper and protection of D. asper resources.

[Dynamics of seed rain of Tripterygium hypoglaucum and soil seed bank].

Tripterygium hypoglaucum is an endangered species in arid areas of Xiannvshan Chongqing, China. The dynamic characteristics of seed rain and soil seed bank of T. hypoglaucum were studied in this paper．Results showed that T. hypoglaucum years of mature seeds distribution number up to October; the seed rain occurred from the last ten-day of September to in the first ten-day of November and the peak of scattered seed rain concentrated in the October．The numbers of soil seed bank at 2-5 cm soil layer,mainly concentrated in the 1.5-3.5 m range. T. hypoglaucum seeds to the wind as a force for transmission, the transmission ability is strong, but in the process of natural reproduction, full mature seed rate is low, the soil seed bank seeds seed short-lived factors these were unfavorable for the natural reproduction of T. hypoglaucum population.

[Analysis on difference of richness of traditional Chinese medicine resources in Chongqing based on grid technology].

After the end of the national and local medicine resources census work, a large number of Chinese medicine resources and distribution of data will be summarized. The species richness between the regions is a valid indicator for objective reflection of inter-regional resources of Chinese medicine. Due to the large difference in the size of the county area, the assessment of the intercropping of the resources of the traditional Chinese medicine by the county as a statistical unit will lead to the deviation of the regional abundance statistics. Based on the rule grid or grid statistical methods, the size of the statistical unit due to different can be reduced, the differences in the richness of traditional Chinese medicine resources are caused. Taking Chongqing as an example, based on the existing survey data, the difference of richness of traditional Chinese medicine resources under different grid scale were compared and analyzed. The results showed that the 30 km grid could be selected and the richness of Chinese medicine resources in Chongqing could reflect the objective situation of intercropping resources richness in traditional Chinese medicine better.

[Studies on difference of chemical compositions in plant species of Tripterygium genus].

We studied the content of chemical compositions and correlation among species of Tripterygium genus by principal component analysis(PCA) and variance analysis(ANOVA), and we also studied the difference among the 3 species.Using [BMIm]PF6 ionic liquid-based ultrasonic-assisted extraction, we determined the contents of 11 compounds including wilforgine, wilforzine, triptophenolide, wilforine, triptoquinone A, triptolide, tripterin, egallocatechin, epigallocatechin, catechin, and epicatechin in 28 batches of the Tripterygium species by HPLC and PCA. Partial least squares analysis (PLS) and ANOVA were also performed to verify the results.The analysis results of PCA and PLS showed that three species of Tripterygium genus were clustered into three regions respectively, and triptoquinone A was the important factor which affected the aggregation of these three species.There was a significant difference among the contents of 11 chemical components in the three species(P<0.000 1).These results indicated that there was a certain correlation between the chemical compositions and the classification of the species, and the difference of the chemical compositions among the three species was obvious. In this work, the content determination method is rapid and accurate, and the analysis method is simple and convenient, which provides a reference for the classification, the efficacy and the toxicity of the species.

Identification of Species in Tripterygium (Celastraceae) Based on DNA Barcoding.

Species of genus Tripterygium (Celastraceae) have attracted much attention owing to their excellent effect on treating autoimmune and inflammatory diseases. However, due to high market demand causing overexploitation, natural populations of genus Tripterygium have rapidly declined. Tripterygium medicinal materials are mainly collected from the wild, making the quality of medicinal materials unstable. Additionally, identification of herbal materials from Tripterygium species and their adulterants is difficult based on morphological characters. Therefore, an accurate, convenient, and stability method is urgently needed. In this wok, we developed a DNA barcoding technique to distinguish T. wilfordii HOOK. f., T. hypoglaucum (LÉVL.) HUTCH, and T. regelii SPRAGUE et TAKEDA and their adulterants based on four uniform and standard DNA regions (internal transcribed spacer 2 (ITS2), matK, rbcL, and psbA-trnH). DNA was extracted from 26 locations of fresh leaves. Phylogenetic tree was constructed with Neighbor-Joining (NJ) method, while barcoding gap was analyzed to assess identification efficiency. Compared with the other DNA barcodes applied individually or in combination, ITS2+psbA-trnH was demonstrated as the optimal barcode. T. hypoglaucum and T. wilfordii can be considered as conspecific, while T. regelii was recognized as a separate species. Furthermore, identification of commercial Tripterygium samples was conducted using BLAST against GenBank and Species Identification System for Traditional Chinese Medicine. Our results indicated that DNA barcoding is a convenient, effective, and stability method to identify and distinguish Tripterygium and its adulterants, and could be applied as the quality control for Tripterygium medicinal preparations and monitoring of the medicinal herb trade in markets.

[Research progress on breeding standard of medicinal animals and discussion on several key problems].

Medicinal animal breeding standards is regarded as the law to normalize relevant production that can guarantee the quality of traditional Chinese medicine of animal category. The article summarized the medicinal animal resources in our country and the present condition of medicinal animal breeding standards. It considered the current animal breeding standards system was in adequate, not only the quantity of breeding standards, the standard content and index were also uncomprehensive, which is not conducive to the scientific and orderly development and utilization of medicinal animal resources. The article pointed out that the development of the basic standards, environmental control, feed quality, raising management, inspection and quarantine should be included into the medicinal animal breeding standards, and the medicinal animal breeding standards content framework was introduced. Meanwhile, animal welfare, biological safety and file management should be concerned during the process of research. Hope the article has good reference value to medicinal animal breeding standards establishment and production management.

[Evaluation of crude drugs by means of colorimeter (I). Study on correlation between color and alkaloids content of coptidis rhizoma].

OBJECTIVE: To examine the correlation between alkaloids content and L*, a* and b* color indices of Rhizoma Coptidis for quality control. METHODS: A colorimeter was used for the measurement of reflected light from sieved powder samples using the CIE 1976 L* a* b* color system. The content of six alkaloids were determined by HPLC. The correlation between alkaloids content and color indices of Rhizoma Coptidis was analyzed. RESULTS: When the particle size of Rhizoma Coptidis was less than 355 microm and the colorimeter parameters were set as measurement diameter of 3 mm, observation degree of 10, and light source of F2 and F7, the measured color was significantly correlated with total alkaloids content (r = 0.793, P < 0.05). As light source of F11, the measured color was significantly correlated with berberine content (r = 0.867, P < 0.01). CONCLUSION: The correlation between the color of powdered Coptidis Rhizoma and its alkaloids contents was found in this study. Measurment of the color of Coptidis Rhizoma can be used to assess its quality.

[Community characters of Lophatherum gracile in Yongchuan district of Chongqing].

To understand Lophatherum gracile plant community's structural characteristics, a survey of community structure and species diversity was conducted through quadrat sampling in Yongchuan district of Chongqing. The results showed that there were 386 species vascular plants, belonging to 117 families and 229 genera. Based on habitat, community structure and species composition, L. gracile were found in three community types: Pinus massoniana community, banboo community, shurb community. Vertical structure was composed of three layers, including tree layer, shrub layer and herb layer. Species in shrub layer was the richness. P. massoniana is the only dominant species of the community, it can not regenerate naturally, the shrub layer has a greater effect on the community of L. gracile in the future. In addition, the banboo community and shurb community is not stable because of human's activity. Therefore, the community characters of L. gracile should be taken care of conservation when the resources are utilized.

Assembly and comparative analysis of the first complete mitochondrial genome of a traditional Chinese medicine Angelica biserrata (Shan et Yuan) Yuan et Shan.

Duhuo, a member of the Angelica family, is widely used to treat ailments such as rheumatic pain. It possesses a diverse array of bioactivities, including anti-tumor, anti-inflammatory, and analgesic properties, as recent pharmacological research has revealed. Nevertheless, the mtDNA of Angelica species remains relatively unexplored. To address this gap, we sequenced and assembled the mtDNA of A. biserrata to shed light on its genetic mechanisms and evolutionary pathways. Our investigation indicated a distinctive multi-branched conformation in the A. biserrata mtDNA. A comprehensive analysis of protein-coding sequences (PCGs) across six closely related species revealed the presence of 11 shared genes in their mitochondrial genomes. Intriguingly, positive selection emerged as a significant factor in the evolution of the atp4, matR, nad3, and nad7 genes. In addition, our data highlighted a recurring trend of homologous fragment migration between chloroplast and mitochondrial organelles. We identified 13 homologous fragments spanning both chloroplast and mitochondrial genomes. The phylogenetic tree established a close relationship between A. biserrata and Saposhnikovia divaricata. To sum up, our research would contribute to the application of population genetics and evolutionary studies in the genus Acanthopanax and other genera in the Araliaceae family.

Tandem Vinylogous Aldol and Intramolecular [2 + 2] Cycloaddition toward Benzocyclobutenes by UV Light Photocatalysis.

A facile and efficient radical tandem vinylogous aldol and intramolecular [2 + 2] cycloaddition reaction for direct synthesis of cyclobutane-containing benzocyclobutenes (BCBs) under extremely mild conditions without using any photocatalysts is reported. This approach exhibited definite compatibility with functional groups and afforded new BCBs with excellent regioselectivity and high yields. Moreover, detailed mechanism studies were carried out both experimentally and theoretically. The readily accessible, low-cost, and ecofriendly nature of the developed strategy will endow it with attractive applications in organic and medicinal chemistry.

Comparative and phylogenetic analysis of Asparagus meioclados Levl. and Asparagus munitus Wang et S. C. Chen plastomes and utility of plastomes mutational hotspots.

Tiandong is a vital traditional Chinese herbal medicine. It is derived from the tuber root of the Asparagus cochinchinensis according to the Pharmacopoeia of the people's republic of China (2020 Edition). On account of the similar morphology, Asparagus meioclados and Asparagus munitus were used as Tian-Dong in southwest China. Chloroplast (cp) genomes are highly active genetic components of plants and play an extremely important role in improving the efficiency of the identification of plant species. To differentiate the medicinal plants belonging to the genus Asparagus, we sequenced and analyzed the complete plastomes (plastid genomes) of A. meioclados and A. munitus and obtained two plastomes whose length changed to 156,515 bp and 156,381 bp, respectively. A total of 111 unique genes have been detected in plastome, which included 78 protein-coding genes, 29 tRNA genes and 4 rRNA genes. In plastomes of A. meioclados and A. munitus, 14,685 and 14,987 codons were detected, among which 9942 and 10,207 had the relative synonymous codon usage (RSCU) values higher than 1, respectively. A. meioclados and A. munitus have 26 SSRs patterns, among which A. meioclados was 25 and A. munitus 21. The average Ka/Ks value was 0.36, and positive selection was detected in genes of the photosynthetic system (ndhF and rbcL) in Asparagus species. To perform the comparative analysis of plastomes, the two newly sequenced plastomes of the A. meioclados and A. munitus species were compared with that of A. cochinchinensis, and 12 hotspots, including 5 coding regions and 7 inter-genomic regions, were identified. Based on the whole plastome of Asparagus, 2 divergent hotspots (accD and rpl32-trnL-UAG) and 1 international barcode fragment (rbcL) were screened, which may be used as particular molecular markers for the identification of Asparagus species. In addition, we determined the phylogenetic relationship between A. meioclados and A. munitus in the genus Asparagus. This study enriches our knowledge of the molecular evolutionary relationships of the Asparagus genus and provides treasured data records for species identification, molecular breeding, and evolutionary analysis of this genus.

[Establishment of seedling classification criteria of Coptis chinensis by dynamic clustering method].

OBJECTIVE: To establish seedling classification criteria of Coptis chinensis. METHOD: The height of plant, leaf number, leaf length, leaf wide, weight of leaf, weight of root were measured, the main measurement indexes of seedlings of C. chinensis were chosen through correlation and regression analysis. The seedling classification criteria were formulated by dynamic clustering analysis. RESULT: The criteria of the 1st-grade seedlings were as follows: leaf number above 8 leaves, height of plant between 12 and 14 cm. The criteria of the 2nd-grade seedlings were as follows: leaf number between 6 and 8 cm, height of plant between 9 and 12 cm. The criteria of the 3rd-grade seedlings were as follows: leaf number between 4 and 6 cm, height of plant above 9 cm. CONCLUSION: The seedling classification criteria of C. chinensis was scientific and feasible, and can be used for the quality control standard of C. chinensis.

The localization of the alkaloids in Coptis chinensis rhizome by time-of-flight secondary ion mass spectrometry.

Background: Understanding the spatial distribution of active compounds can effectively evaluate the quality of decoction pieces of traditional Chinese medicine (TCM). Traditional methods are economical and practical but lack chemical information on the original distribution. Time-of-flight secondary ion mass spectrometry (TOF-SIMS), with the advantage of non-destructive detection of samples, can directly analyze the distribution of chemical compounds on the surface of various samples. Methods: In this study, TOF-SIMS image analysis technology was used to detect TCM for the first time. Taking Coptis rhizome (CR) as an example, a commonly used TCM, the distribution of the compounds in the cross-section of CR was studied. Meanwhile, ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLCQQQ-MS/MS) was used to verify the results of TOF-SIMS. Results: The distribution of nine active compounds: berberine, epiberberine, coptisine, palmatine, columbamine, jatrorrhizine, tetrahydricheilanthifolinium, and oxyberberine, was well imaged in the cross-section of CR by TOF-SIMS. The content of berberine and epiberberine was the highest; Palmatine distribution in the pith was more than that in other parts; Oxyberberine was mainly concentrated in the cork and xylem rays. Normalization analysis showed contents of these compounds increased along with the growth years. The result was consistent with UPLC-QQQ-MS/MS. Conclusion: The TOF-SIMS method can display the spatial distribution status of the active compounds of herbs, providing a basis for selecting the medicine site with non-destructive and fast detection.

[Study on the net photosynthesis rate of Coptis chinensis from different types and production places].

OBJECTIVE: Evaluate the photosynthesis characteristics of Coptis chinensis and provide theoretical basis for the optimization of Coptis chinensis germplasm resources and variety. METHODS: The daily variation of the net photosynthetic rate, transpiration rate, stomatal conductance and intercellular CO2 density in leaf blade were determined by Photosynthesis analyzer, and the net photosynthesis rate of Coptis chinensis from different plant ages, types and production places was compared. RESULTS: The light saturation point in Coptis chinensis was about 500 micromol/m2 x s, light compensation point was about 12.04 micromol/(m2 x s), apparent quantum yield was 0.011. When the temperature was above 33 degrees C, the transpiration rate increased but the net photosynthesis rate decreased. There was no significant difference (P > 0.05) among the net photosynthesis rate of Coptis chinensis from different plant ages. The net photosynthesis rate of the type DA-YE and ZHI-HUA was significantly higher than that of other types (P < 0.05); The net photosynthesis rates of specimens from Fubao mountain in Lichuan county of Hubei and Fengmu village in Shizhu county of Chongqing were significantly higher than that of others. CONCLUSIONS: The net photosynthesis rate of Coptis chinensis decreased under hight temperation and hight light. Different germplasm resources of Coptis chinensis, the net photosynthetic rate had certain differences.

[Dynamic accumulation of effective components and biomass of Coptis chinensis in Hongya county].

OBJECTIVE: To study the dynamic accumulation of the effective components and biomass of Coptis chinensis, so to provide the experimental date of optimal harvest time for C. chinensis in Hongya county. METHOD: The samples of three to five years were gathered from the same field and time. The biomass was analyzed by weighed. The jatrorrhizine, columbamine, epiberberine, coptisine, palmatine and berberine in C. chinensis were analyzed by HPLC. RESULT: With the increasing of years of growth, the rootstalk biomass of C. chinensis was increasing continuously. The biomass growth of four-year-old C. chinensis was the fastest in the year. From September to October was the fastest season of the growth of rootstalk. The dynamic accumulation in rootstalk C. chinensis had regularity in the certain extend. The contents of six alkaloids and all alkaloids in 4-year-old C. chinensis were more than that in 3-years-old and 5-year-old. The contents of six alkaloids were mostly highest in August. From July to December, there is no significant difference in the contents of columbamine, epiberberine, coptisine, palmatine, berberine and all alkaloids in 4-years-old C. chinensis. CONCLUSION: According to the biomass and the accumulation pattern of the effective components in the C. chinensis, the optimal harvest time is from September to October of 4-year-old C. chinensis.

The complete chloroplast genome of Saussurea medusa maxim. (Asteraceae), an alpine Tibetan herb.

Saussurea medusa is an important traditional Tibetan medicinal plant in China. In this study, we assembled the complete chloroplast (cp) genome of S. medusa. The complete S. medusa chloroplast genome is a circular molecular structure of 152,257 bp in length with coding GC 37.93%, consisting of two inverted repeats (25,204 bp) separated by a large single-copy region (83,334 bp) and a small single-copy region (18,515 bp). The complete chloroplast genome of S. medusa contained 130 genes, including 87 protein-coding genes, 35 tRNA genes, and eight rRNA genes. Phylogenetic analysis shows that S. medusa is most closely related to Saussurea inversa and Saussurea pseudoleucoma. The complete chloroplast genome sequence of S. medusa facilitates the phylogenetic studies of Asteraceae.

[Determination of 2,3,5,4'-tetrahydroxystilbene-2-O-beta-D-glucopyranoside and anthraquinon simultaneously in Polygonum multiflori of different growing stage and gathering periods by HPLC].

OBJECTIVE: A HPLC method was established for the determination of 2, 3, 5, 4'-tetrahydroxystilbene-2-O-beta-D-glucopyranoside and anthraquinon simultaneously in Polygonum multiflori. The contents variation from different growing stages in P. multiflori were observed. METHOD: The HPLC method was used to measure the contents of 2, 3, 5, 4'-tetrahydroxystilbene-2-O-beta-D-glucopyranoside and anthraquinon simultaneously in 14 years old P. multiflori which were harvested in July to December. Chromatographic conditions: The chromatography column was C18 with a mobile phase composed of methanol-0.1% water phosphoric acid. The detection wavelength was 320 nm and 254 nm respectively, and the flow rate was 1.0 mL x min(-1). RESULT: In a certain range, 2, 3, 5, 4'-tetrahydroxystilbene-2-O-beta-D-glucopyranoside, emodin and physcion showed good linearity, precision, reproducibility, stability and average recovery. CONCLUSION: The developed method is sensitive, accurate. The contents of 2, 3, 5, 4'-tetrahydroxystilbene-2-O-beta-D-glucopyranoside and anthraquinon in P. multiflorum rapidly accumulated in one to two years, and in three year that was the highest. The samples collected in November accumulate much higher amounts of 2, 3, 5, 4'-tetrahydroxystilbene-2-O-beta-D-glucopyranoside and anthraquinon than those which were collected in other months.