Herbal Textual Research on Alpiniae Officinarum Rhizoma in Famous Classical Formulas / 中国实验方剂学杂志

By reviewing ancient materia medica， medical books and modern literature， this paper conducted a systematic research on name， origin， scientific name evolution， producing area， quality， harvesting and processing methods of Alpiniae Officinarum Rhizoma. The results showed that Alpiniae Officinarum Rhizoma was first published in Mingyi Bielu， and its correct name was Gaoliangjiang. The mainstream origin of Alpiniae Officinarum Rhizoma used in the past dynasties is Alpinia officinarum， which is used to this day， while it used to be mixed with A. galanga because of the similar name and morphology. Alpiniae Officinarum Rhizoma produced in Danzhou and Leizhou was considered to be better in ancient times， and now it mainly produced in Guangdong， Guangxi and Hainan provinces. In addition， it has been concluded that Alpiniae Officinarum Rhizoma with reddish brown， sturdy and firm character， wrinkled skin， convex flesh， aromatic and spicy taste， and few branches is the best. In ancient times， Alpiniae Officinarum Rhizoma was commonly harvested in February and March， whereas it generally harvested in late summer or early autumn at present， and wild products are usually harvested before the rainy season in May. The main processing methods of Alpiniae Officinarum Rhizoma are cleansing and cutting， and some other methods are stir-frying or mixing with auxiliary materials. Based on the research results， it is suggested that the raw products of A. officinarum rhizomes or its processed products according to prescription requirements should be used in the development of famous classical formulas containing Alpiniae Officinarum Rhizoma.

The Development Strategy and Mode of Co-construction and Sharing Base to Improve the Chinese Herbal Medicine Quality / 世界科学技术-中医药现代化

Chinese medicinal materials occupy an important position in China's health industry. However, its overall quality needs to be improved and it is in urgent need of regulation. Exploring the formation of effective organizational mechanisms and industry models has become an urgent need of the industry. In this context, the alliance of coconstruction and sharing Chinese herbal medicine base came into being. The alliance is based on the pursuit of the quality of Chinese herbal medicines and continues to promote the construction of Chinese herbal medicines. The Alliance provides a platform for economic and scientific cooperation in the industry. Its purpose is to guide the promotion of the standardization of local varieties and the construction of modern Chinese medicine agricultural enterprises based on the development needs of Chinese herbal medicine resources and the common interests of all members. As an important content, we will strive to expand the new pattern of coordinated development of traditional Chinese medicine agriculture and industry, explore the establishment of a new organizational system for modern Chinese medicine agricultural production with controllable quality, output and price under the link of production and demand. For the sustainable, stable and healthy development of the Chinese medicine industry, it will serve 1.3 billion people and serve humanity, provide high-quality sustainable Chinese herbal medicine resources. Since its establishment six years ago, the alliance has carried out work on key aspects such as standardized production of Chinese herbal medicines, plant protection, decoction processing, supply and demand docking, medicinal materials standards, poverty alleviation, breeding, and provided technical support to enterprises. During this period, the alliance also proposed the concept of"three-no and one- all"requires the members to take the lead in achieving the standards of"sulfur- free processing, no aflatoxin pollution, pollution-free, traceable throughout the whole", setting a benchmark for the industry.

Application of CRISPR/Cas9 in plant biology

The CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins) system was first identified in bacteria and archaea and can degrade exogenous substrates. It was developed as a gene editing technology in 2013. Over the subsequent years, it has received extensive attention owing to its easy manipulation, high efficiency, and wide application in gene mutation and transcriptional regulation in mammals and plants. The process of CRISPR/Cas is optimized constantly and its application has also expanded dramatically. Therefore, CRISPR/Cas is considered a revolutionary technology in plant biology. Here, we introduce the mechanism of the type II CRISPR/Cas called CRISPR/Cas9, update its recent advances in various applications in plants, and discuss its future prospects to provide an argument for its use in the study of medicinal plants.

Research Progress in Cymbopogon citratus (DC.) Stapf and Lemongrass Tea / 世界科学技术-中医药现代化

C.citratus has been usedin many countries with a long history.Traditionally,it is applied as a food seasoning in cooking.It is also used in tea beverage and folk medicine as well.Modern application of C.citratus is focused on the development of citronella oil,which can be used for food additives,disinfectants,cosmetics,drugs and etc.C.citratus is also a potential plant in landscaping.Its special lemony flavor contains chemical constituents,mainly including citral,myrcene,linalool,geraniol,nerol,citronellol,and etc.The modern research showed that C.citratus had the main effects of anti-microbial,anti-inflammation,analgesia,anti-oxidation,anti-tumor,anti-anxiety,anti-hypertension,antihyperglycemia,and etc.With further studies,some new pharmacological properties of C.citrates are going to be discovered gradually.It is worthy of further research and development to meet the needs of the health industry.

Syringaresinol-4---d-glucoside alters lipid and glucose metabolism in HepG2 cells and C2C12 myotubes

Syringaresinol-4---d-glucoside (SSG), a furofuran-type lignan, was found to modulate lipid and glucose metabolism through an activity screen of lipid accumulation and glucose consumption, and was therefore considered as a promising candidate for the prevention and treatment of metabolic disorder, especially in lipid and glucose metabolic homeostasis. In this study, the effects of SSG on lipogenesis and glucose consumption in HepG2 cells and C2C12 myotubes were further investigated. Treatment with SSG significantly inhibited lipid accumulation by oil red O staining and reduced the intracellular contents of total lipid, cholesterol and triglyceride in HepG2 cells. No effect was observed on cell viability in the MTT assay at concentrations of 0.1-10 μmol/L. SSG also increased glucose consumption by HepG2 cells and glucose uptake by C2C12 myotubes. Furthermore, real-time quantitative PCR revealed that the beneficial effects were associated with the down-regulation of sterol regulatory element-binding proteins-1c, -2 (), fatty acid synthase (), acetyl CoA carboxylase () and hydroxyl methylglutaryl CoA reductase (), and up-regulation of peroxisome proliferator-activated receptors alpha and gamma (and). SSG also significantly elevated transcription activity oftested by luciferase assay. These results suggest that SSG is an effective regulator of lipogenesis and glucose consumption and might be a candidate for further research in the prevention and treatment of lipid and glucose metabolic diseases.

Effect of agarwood produced by whole-tree agarwood-inducing technique on hypnotic and spontaneous activity inhibition of mice / 国际药学研究杂志

Objective To evaluate the sedative-hypnotic effect of agarwood different extracts produced by whole-tree agar?wood-inducing technique(Agar-Wit agarwood)to make scientific basis for the Agar-Wit agarwood exploration and usage in clinic. Method Collaborative pentobarbital sodium hypnosis experiment was employed to assess the index of fell asleep rate ,sleep latency time and sleep time. And locomotor activity assay was used to test the index of moved distance,travel time and average velocity. Then a functional comparison was made between Agar-Wit agarwood and commereial agarwood. Result The Agar-Wit agarwood ethanol ex?tract and essential oil could significantly increase sleep rate and prolong sleep time. At the same time ,essential oil could also decrease sleep latency time significantly. Additionally,both ethanol extract and oil significantly decreased the mice locomotor activity,includ?ing reduced total distance,movement,move time and average velocity. However,the water extract did not have significant effect. Conclusion The Agar-Wit agarwood ethanol extract and oil have significant effect on sedation in mice,whose function is similar to,or even better than that of the commerical agarwood.

Effect of agarwood extracts produced by the whole-tree agarwood-inducing technique on gastrointestinal motility and gastric ulcer / 国际药学研究杂志

Objective To evaluate the gastrointestinal regulative effect of agarwood extracts produced by the whole-tree agar?wood-inducing technique(Agar-Wit agarwood)to make scientific basis for the Agar-Wit agarwood exploration and usage in clinic. Methods Intestinal propelling effect assay marked by carbon powder,and gastric empting effect assay marked by methyl orange were used to evaluate the gastrointestinal effects of Agar-Wit agarwood on mice by single or constant repeated intragastric(ig)adminis?tration of water extract and ethanol extract,and then the index of intestinal peristaltic rate and methyl orange relative residual rate were calculated. Gastrtic ulcer model was established under water stress to appraise their protective function on rat stomach and to ob?tain the gastric ulcer index and inhibitory rate. Results The Agar-Wit agarwood ethanol extract significantly improved intestinal peri?stalsis and gastric empting function by single or constant repeated ig administration at the dose of 150 mg/kg. The ethanol extract of commercial agarwood also had the similar effect at the dose of 450 mg/kg. But the water extract did not have significant effect. The gas? tric ulcer assay results showed that the ethanol extract significantly inhibited gastric ulcer happening by single ig administration. Addi?tionally,constant repeated ig administration of ethanol extract showed the consistent results and the gastric ulcer inhibitory rate was (73.1±5.6)%at the dose of 300 mg/kg. Conclusion The Agar-Wit agarwood ethanol extracts have significant intestinal peristaltic ef?fect,gastric empting effect and gastric ulcer inhibitory function.

Cloning, expression and characterization of COI1 gene (AsCOI1) from Aquilaria sinensis (Lour.) Gilg

Aquilaria sinensis, a kind of typically wounding-induced medicinal plant with a great economical value, is widely used in the production of traditional Chinese medicine, perfume and incense. Coronatine-insensitive protein 1 (COI1) acts as a receptor in jasmonate (JA) signaling pathway, and regulates the expression of JA-responsive genes in plant defense. However, little is known about the COI1 gene in A. sinensis. Here, based on the transcriptome data, a full-length cDNA sequence of COI1 (termed as AsCOI1) was firstly cloned by RT-PCR and rapid-amplification of cDNA ends (RACE) strategies. AsCOI1 is 2330 bp in length (GenBank accession No. KM189194), and contains a complete open frame (ORF) of 1839 bp. The deduced protein was composed of 612 amino acids, with a predicted molecular weight of 68.93 kDa and an isoelectric point of 6.56, and was predicted to possess F-box and LRRs domains. Combining bioinformatics prediction with subcellular localization experiment analysis, AsCOI1 was appeared to locate in nucleus. AsCOI1 gene was highly expressed in roots and stems, the major organs of agarwood formation. Methyl jasmonate (MeJA), mechanical wounding and heat stress could significantly induce the expression level of AsCOI1 gene. AsCOI1 is an early wound-responsive gene, and it likely plays some role in agarwood formation.

Establishment of a cell suspension culture system of endangered Aquilaria sinensis (Lour.) Gilg / 药学学报

Aquilaria sinensis callus induced by stem tips were used to establish the suspension cell system. The results showed that the most suitable medium for callus induction and subculture is MS + 2.0 mg x L(-1) NAA + 1.0 mg x L(-1) 6-BA. After 12 times of subculture, the energetic and loose callus, which were appropriate for cell suspension culture, were cultured and shook in liquid medium MS + 2.0 mg x L(-1) NAA + 1.0 mg x L(-1) 6-BA + 500.0 mg x L(-1) casein hydrolysate (CH) to establish the suspension cell system. The growth curve of suspension cells showed a "S" type. At the beginning of the culture, cell density increased slowly; during 4 to 6 days, suspension cells reached logarithmic growth period; during 7 to 12 days, suspension cells were in the platform period; but after 12 days, cell density and activity went down obviously. Agarwood sesquiterpenes were not detected in the suspension cells during the growth period, however, they could be detected in MeJA treated suspension cells. In this study, a stable and active growing suspension cell system was established, which was a proper system to study the mechanism of agarwood sesquiterpene formation, and additionally provided a potential way to generate agarwood sesquiterpenes through application of cell culture.

Cloning and expression analysis of HMG-CoA reductase from Aquilaria sinensis (Lour.) Gilg / 药学学报

3-Hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) is the first rate-limiting enzyme for sesquiterpene synthesis in the mevalonate (MVA) pathway. The specific primers were designed according to the transcript sequence of AsHMGR2 from the Aquilaria sinensis (Lour.) Gilg transcriptome database. The full-length cDNA of AsHMGR2 was cloned by RT-PCR and rapid amplification of cDNA ends (RACE) technology, and was analyzed at bioinformatics levels; AsHMGR2 expression profiles in different tissues and in responds to different treatments were analyzed by real-time PCR. The length of AsHMGR2 Open Reading Frame (ORF) was 1 749 bp, encoding 582 amino acids. The GenBank accession number is KC140287. Tissue expression analysis indicated that AsHMGR2 was mainly expressed in root and shoot tips, followed by stem, and was lowest in leaves. Inducible-experiments showed that the genes were induced by mechanical wound as well as chemical liquid induction, and reached the highest expression level at 6 h and 8 h, separately. The full-length cDNA of AsHMGR2 and its expression patterns will provide a foundation for further research on its function in agarwood sesquiterpene biosynthesis.

Study on changes and influence of humoral immunity in premature infants / 重庆医学

Objective To study changes of humoral immunity of the premature infants in different pathological conditions and detect the reason of the deficiency of humoral immunity in premature infants .Methods Two hundred and forty-six prematur were enrolled and 30 healthy neonates were selected as control group .The percentages of IgG ,IgA ,IgM and comp lement C3 ,C4 were detected by full automatic biochemical analyzer .Results The results showed that IgG ,IgM ,IgA ,C3 and C4 in the premature in-fants were lower than those in the normal term infants and there was a highly significant difference with the decrease of fetal age . IgG ,IgM ,IgA ,C3 and C4 of the group of the premature infants ranging from 32 to 36 weeks had reduced in different degree ,rela-tive to the groups of BW <2 000 g ,hypertension during pregnancy ,cesarean section(P<0 .05) .Conclusion The results showed that function of humoral immunity in the premature infants was depressed and low gestational age ,low birth weight ,cesarean sec-tion and hypertension during pregnancy may be the leading cause of the deficiency of humoral immunity .

Expression analysis of glycosyltransferase BcUGT1 from Bupleurum chinense DC. and its expression in E. coli and the target protein purification / 药学学报

The ORF sequence of glycosyltransferase gene BcUGT1 cloned from Bupleurum chinense DC. was analyzed and its three dimentional structure was predicted. Using qRT-PCR method, the expression characteristics of BcUGT1 after methyl jasmonate (MeJA) induction and in different plant tissues were investigated. The results showed that BcUGT1 may be involved in saikosaponin biosynthesis in B. chinense. Thereafter, the recombinant vectors of BcUGT1 were constructed for its expression in E. coli. The target protein was successfully expressed and purified. In the present study, three vectors, pRSET-A, pET-28a (+) and pET-30a (+), and three isolates of E. coli, BL21 (DE3) plysS, BL21A1 and BL21-CodonPlus (DE3)-RIPL were used under different induction conditions, such as different concentrations and during times of inducers (L-arabinose and IPTG) and different inducing temperatures. The results showed that in the condition of 0.5 or 1 mmol x L(-1) IPTG, 16 degrees C, 20 h, target protein expressed in BL21-CodonPlus (DE3)-RIPL with pET-28a (+) or pET-30a (+) as vector. Using PrepEase His-tagged protein purification kit, the target protein was purified. The present work will be helpful for follow-up bio-function analysis of BcUGT1.

Induction of hairy roots and plantlet regeneration of Bupleurum chinense DC / 药学学报

In this study, the induction of hairy roots of Bupleurum chinense DC. was explored and established after experiments at different conditions: A. rhizogenes A4 was used to infect the leaves bases of B. chinense tube seedlings. The explants were co-cultured on Phytagel-solidified media for 3 days and then, were turned into solid media, similar with the co-culture media except that bacteriostat was added. After 10 days, rootlets began to appear and after 4 to 5 weeks, rootlets can be converted into liquid shaking culture stage. Plants regeneration from hairy root was useful for the research of new germplasm production and the variety improvement breeding. In the present study, the regenerated plants were obtained. One approach was to continuously culture under light conditions the seedlings which parting off spontaneously from the hairy roots during liquid shaking culture. The other approach was to culture the callus-like tissues produced by hairy roots with the optimized regeneration media for the induction of regenerated plants. The results of present study provide a technique to induce hairy roots and plantlet regeneration of B. chinense and this technique is helpful for the researches on metabolism, especially on the Agrobacterium-mediated genetic transformation of B. chinense.

Cloning of UGT gene of Bupleurum chinense and construction of over expressing and RNAi transgenic vectors / 中国中药杂志

<p><b>OBJECTIVE</b>To clone the full-length cDNA of a uridine diphosphate glycosyltransferase (UGT) gene which may be involved in saikosaponin biosynthesis of Bupleurum chinense, and construct the transgenic vectors for over expression and RNAi of the cloned UGT. These works will provide foundation for further its function analysis by transgene study.</p><p><b>METHOD</b>RAGE and LD-PCR were used to clone the full-length cDNA of the UGT, on the basis of its partial cDNA sequence obtained from our previous 454-sequenced dataset. The ORF and partial sequences of the UGT were PCR cloned using primers with corresponding restriction enzymes cutting sites. The PCR products were digested with corresponding restriction enzymes and then were inserted into pCAMBIA-SUPER 1 300 and pHANNIBAL. The recombinant pHANNIBAL were digested with Not I and then were inserted into a binary vector, pART27. Finally, the transgenic vectors for over expression and RNAi of the cloned UGT were constructed.</p><p><b>RESULT</b>The full-length cDNA of a UGT were cloned from B. chinense. The recombinant vectors for over expression and RNAi of the UGT were obtained.</p><p><b>CONCLUSION</b>Our works on full-length cDNA cloning and transgenic vectors construction provide a substantial foundation for follow-up biofunction analysis of the UGT through transgenic research.</p>

Factors Affecting Embryogenic Callus Production and Plant Regeneration in Anther Culture of Bupleurum chinense / 中草药·英文版

Objective To evaluate the influences of the genotypes,anther developmental stages,and cultural conditions on the efficiency of embryogenic callus induction and plant regeneration in the anthers culture of Bupleurum chinense.Methods The different effects such as four genotypes,plant growth regulators,and temperature condition were compared in the experiments.The histological study was performed with the process of the anther culture.Results The highest inducing rate of embryogenic calli were achieved for the genotypes Zhongcaiyihao(ZCYH),Z4,and Z5 at the early-to middle-uninucleate stages,except for genotype ZPM1 at the tetrad stage.Cold pretreatment increased the production of the embryogenic callus,in which 4-day cold pretreatment improved the production of embryogenic callus from 0% to 2.2% and 5.0% for genotypes ZPM1 and ZCYH,respectively.No embryogenic callus was induced in the medium containing less than 0.75 mg/L 2,4-dichlorophenoxyacetic acid(2,4-D).The highest regeneration rate (34.6%)was obtained in 1/2 MS salts regeneration medium supplemented with 0.1 mg/L 6-benzylmaminopurine (BA).The low concentration of BA was able to promote the embryogenic callus formation and subsequent plantlet regeneration via somatic embryogenesis.Chromosome counting of regenerated plantlets showed mostly diploid plant (2n = 12)with only one haploid plant(n = 6).Because of the low rate of microspore embryo formation,we only tracked the process of embryogenesis from the connective tissue,instead of microspore by histological observations.Conclusion This study establishes an efficient system for embryogenic callus induction and plant regeneration system.This is the first report on the haploid plantlet through the anther culture orB.chinense.

Characteristics and abortion cytological pathways of male sterility of Bupleurum chinense / 中国中药杂志

<p><b>OBJECTIVE</b>To study the characteristics of male sterility of Bupleurum chinense and further explore the developmental period and reason of abortion for the male sterile plants.</p><p><b>METHOD</b>The morphological characteristics of B. chinense male sterile and normal plants were investigated and compared. The anther development process and pollen viability of two types of plants were examined by microscopic assay.</p><p><b>RESULT AND CONCLUSION</b>The shapes and sizes of anther and filament were different between the male sterile and the normal plants. For the male sterile plant's, the filament size was no more than 1/2 of that of normal plants and the anthers were shriveled, failed to dehisce and pollinate naturally, and the pollen grains in the anthers had no vitality. Other morphological characteristics were similar between two types of plants. The main reason leading to male sterility of B. chinense was the abnormal development of tapetum cells with two circumstances. The one is that the tapetum cells degraded early during the period of microsporocyte phase to tetrad phase and the other is that the tapetum cells proliferated with delayed degradation in the tetrad to uninucleate phase.</p>

Breeding of new varieties "zhongchai no. 2" and "zhongchai no. 3" of Bupleurum chinense / 中国中药杂志

<p><b>OBJECTIVE</b>To breed new varieties with better uniformity and commercial quality as well as higher saikosaponin contents.</p><p><b>METHOD</b>The excellent germplasm resources were selected from "zhongchai no. 1" population. Single plant method was applied to get better varieties. All the breeding material was investigated according to morphological characters, agronomic characters and the contents of saikosaponin a and saikosaponin d. The experiments of comparative test and varieties regional test were carried out.</p><p><b>RESULT</b>The bred new varieties of "zhongchai No. 2" and "zhongchai No. 3" had better uniformity. The dark brown roots ratios of the two varieties were 83.2%, 89.9%, respectively. The contents of saikosaponins (a + d) of the two varieties reached 1.31%, 1.02%, respectively.</p><p><b>CONCLUSION</b>"zhongchai No. 2" and " zhongchai No. 3" both had the advantages of better uniformity, darker brown roots and higher saikosaponin contents.</p>

Floral dynamic and pollination habit of Fritillaria cirrhosa / 中国中药杂志

<p><b>OBJECTIVE</b>To find the sexual reproduction characteristics and improve the seed propagation and new variety breeding of Fritillaria cirrhosa.</p><p><b>METHOD</b>Flower, anther and pistil development were observed. Pollinating in different development period and bagging were used to measure stigma maturity situation and natural outcrossing rate.</p><p><b>RESULTS AND CONCLUSION</b>It took 12 days from floral bud emergency to finished flowering. It was observed that anther opened in longitudinal direction and pollen was ejected for 2-4 days continuously. Pistil matured earlier, and chapiter could be fertilized from middle bud stage the third day after flowering, but the most suitable time was 2-3 day after the corolla opening. The natural outcrossing rate was 81.9%. F. cirrhosa is a typical xenial plant.</p>

Construction of genetic linkage map of Bupleurum chinense DC. using ISSR and SSR markers / 药学学报

Abstract: Molecular genetic map is a fundamental organizational tool for genomic research. However, a genetic linkage map for Bupleurum chinense DC. has not been developed. In this study, with the theory of pseudo-testcross, 96 F1 plants from an intraspecific cross of B. chinense were used as mapping populations. Twenty eight ISSR (inter-simple sequence repeat) primers and 44 SSR (simple sequence repeat) primers were used to detect the polymorphisms between the parental plants, and of them, 28 ISSRs and 14 SSRs were selected to analyze the F1 populations. The map consisted of 13 linkage groups which included 80 (72 ISSRs and 8 SSRs) loci, and covered 2 633.9 cM with an average density of 33.4 cM. All 13 linkage groups consisted of 2-31 loci ranging in length from 15.4-1295.7 cM. This map will provide a basis for studies on gene mapping, map-based cloning and maker-assisted selection of important traits in B. chinense.

Comparison on agronomy and quality characters of selective strain of Schizonepeta tenuifolia / 中国中药杂志

<p><b>OBJECTIVE</b>With the purpose of selecting adequate quality and high production of Schizonepeta tenuifolia, the comparative experiments were carried out on different strain of S. tenuifolia in 2007.</p><p><b>METHOD</b>The test fields were divided into blocks randomly, and the agronomy characters were investigated in harvest time; the content of volatile oil was measured by steam distillation and the pulegone were determined by HPLC.</p><p><b>RESULT</b>The yield of S4 was 18.63% and 29.99% higher than that of CK1 and CK2, respectively. The contents of volatile oil and pulegone were also higher than those of CK and other strains in this test.</p><p><b>CONCLUSION</b>S4 shows the advantages of high production, strong disease resistance and high active components. S4 would be extended as the good breed in production.</p>