The Role of Traditional Chinese Formula Ding-Kun Pill (DKP) in Expected Poor Ovarian Response Women (POSEIDON Group 4) Undergoing In Vitro Fertilization-Embryo Transfer: A Multicenter, Randomized, Double-Blind, Placebo-Controlled Trial.

Objective: The primary objective of the study was to assess traditional Chinese formula DKP supplementation in terms of efficacy and safety on reproductive outcomes of expected poor ovarian responder (POR, POSEIDON Group 4) undergoing in vitro fertilization-embryo transfer (IVF-ET). Design Setting and Participants: Women eligible for IVF-ET were invited to participate in this randomized, double-blind, placebo-controlled, superiority trial at academic fertility centers of ten public hospitals in Chinese Mainland. A total of 462 patients (35-44 years) equally divided between DKP and placebo groups with antral follicle count (AFC) <5 or anti-müllerian hormone (AMH) <1.2 ng/ml were randomized. Interventions: All participants were given DKP or 7 g placebo twice daily on the previous menstrual cycle day 5 until oocyte retrieval, which took approximately 5 to 6 weeks. Main Outcome Measure: The primary outcome was the ongoing pregnancy defined as more than 20 gestational weeks of an intrauterine living fetus confirmed by pelvic ultrasonography. Results: Demographic characteristics were equally distributed between the study populations. Intention-to-treat (ITT) analysis revealed that ongoing pregnancy rate (OPR) was not significantly different between DKP and placebo groups [26.4% (61/231) versus 24.2% (56/231); relative risk (RR) 1.09, 95% confidence interval (CI) 0.80 to 1.49, P = 0.593]. No significant differences between groups were observed for the secondary outcomes. The additional per protocol (PP) analysis was in line with ITT results: OPR in DKP group was 27.2% (61/224) versus 24.1% (55/228) in placebo group [RR 1.13, 95%CI (0.82 to 1.55), P = 0.449]. After subgroup analysis the findings concluded that POR population of 35-37 years had a significantly higher OPR after 5-6 weeks of oral DKP (41.8%, 33/79) versus placebo (25.4%, 18/71) [RR 1.65, 95% CI (1.02 to 2.65), P = 0.034, P for interaction = 0.028]. Conclusion: This well-designed randomized controlled trial (RCT) offers new high-quality evidence to supplement existing retrospective literature concerning DKP performance in expected PORs. DKP could be recommended as a safe and natural remedy for expected PORs (aged 35-37 years) who fulfill the POSEIDON group 4 criteria. However, additional interventional clinical studies are undoubtedly required to be conducted in the future to validate this hypothesis. Clinical Trial Registration: www.chictr.org.cn, identifier ChiCTR1900026614.

[Preliminary research of deqi and the tissue depth of acupoint thread embedding therapy on weight reduction under ultrasonic guidance].

OBJECTIVE: To explore the impacts on weight reduction effect treated with acupoint thread embedding therapy at different tissue levels under ultrasonic guidance. METHODS: A total of 70 patients with overweight or obesity were randomized into a shallow-tissue thread embedding group (35 cases, 5 cases dropped off) and a deep-tissue thread embedding group (35 cases, 4 cases dropped off). Under ultrasonic guidance, the thread was embedded in the shallow tissue level and the deep tissue level respectively. The acupoints were Zhongwan (CV 12), Xiawan (CV 10), Shuifen (CV 9), Zhongji (CV 3), etc. The thread embedding therapy was exerted once every 2 weeks, totally for 3 times. Before and 2 weeks after treatment, body mass, body mass index (BMI), waist circumference and hip circumference were recorded in the patients of the two groups separately. After each treatment, the number and the property of blood vessels under each acupoint were detected by ultrasound. Besides, the needling sensation and the intensity were scored and the adverse events were observed after thread embedding therapy. RESULTS: After treatment, the reduction range of body mass, BMI and waist circumference in the deep-tissue thread embedding group were larger than those in the shallow-tissue thread embedding group successively (P<0.05). The scores of distention and fullness sensation, needling sensation and intensity in the deep-tissue thread embedding group were higher than those in the shallow-tissue thread embedding group successively (P<0.05). Of 29 cases in the shallow-tissue thread embedding group and 27 cases of the deep-tissue thread embedding group, under at least one acupoint, the vessels were found and distributed unevenly (P<0.05). There were no adverse events, i.e. bleeding and infection in the two groups. CONCLUSION: The deep-tissue thread embedding therapy achieves the stronger deqi (needling sensation) and better effect of weight reduction. The acupoint thread embedding therapy under ultrasonic guidance can accurately locate the tissue depth and reduce the incidence of adverse events of thread embedding treatment.

Nocardia panacis sp. nov., a novel actinomycete with antiphytopathogen activity isolated from the rhizosphere of Panax notoginseng.

Strain YIM PH21724T was isolated from the rhizosphere of Panax notoginseng. Phylogenetic analyses based on 16S rRNA gene sequences showed that the strain exhibits close phylogenetic relatedness to Nocardia kroppenstedtii N1286T (97.70%), Nocardia farcinica NCTC 11134T (97.67%) and Nocardia puris DSM 44599T (97.40%). The menaquinones were identified as MK-9 (H4), MK-8 (H4, ω-cyclo) and MK-8 (H4), and the major fatty acids (> 10%) were identified as C16:0, C18:1 ω9c and C18:0 10-methyl. The polar lipids were found to be composed of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannosides and an unidentified lipid. The G + C content of the genomic DNA was determined to be 67.01 mol%. The phenotypic, chemotaxonomic, phylogenetic and genomic results clearly show strain YIM PH21724T should be classified in the genus Nocardia and represents a novel species, for which the name Nocardia panacis sp. nov. is proposed. The type strain is YIM PH21724T (= DSM 105904T = KCTC 49030T = CCTCC AA 2017043T).

Simultaneous analysis of 23 illegal adulterated aphrodisiac chemical ingredients in health foods and Chinese traditional patent medicines by ultrahigh performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry.

This paper presents an application of ultra high-performance liquid-chromatography-quadrupole-TOF high resolution mass spectrometry (UHPLC-Q-TOF HRMS) for simultaneous analysis of 23 illegal adulterated aphrodisiac type chemical ingredients in health foods and Chinese Traditional Patent Medicines (CTPMs). The mass spectrometer was operated in Information Dependent Acquisition (IDA) mode, which provides crucial information for the elemental composition analysis, structure elucidation and quantitative analysis simultaneously. Quantitative analysis was performed using the peak areas of the precursor ions in the XICs. The method validation included assessment of selectivity, sensitivity, calibration curve, accuracy, precision, recovery, matrix effect and stability. The results show good linear relationship with the concentrations of the analytes over wide concentration ranges (e.g., 0.05-10 µg/g for sildenafil) as all the fitting coefficients of determination r2 are >0.9984. The detection limits (LODs) were in the range of 0.002-0.1 µg/g. The recoveries were able to reach 82.5-103.6%, while the matrix effects ranged from 87.7 to 109.3%. The intra- and inter-day accuracies were in the range of 82.3-113.8%, while the intra- and inter-day precision ranged from 0.4 to 13.6%. Among 40 batches of health foods and 32 batches of CTPMs (including 28 capsules, 32 tablets, 10 liquid and 2 pills) samples, 28 batches of heath foods were positive. The detected chemical ingredients involved sildenafil, tadalafil, aildenafil and sulfoaildenafil. This method can be used for the screening, identification and quantification of illegal adulterated aphrodisiac chemical ingredients in health foods and CTPMs. Moreover, the LC-Q-TOF MS is very useful to structural elucidation of unknown compound.

Ligustilide Relieves Complete Freund's Adjuvant-induced Mechanical Hyperalgesia through Inhibiting the Activation of Spinal c-Jun N-terminal Kinase/c-Jun Pathway in Rats.

BACKGROUND: Ligustilide, an active ingredient in a traditional Chinese medicine, has anti-inflammatory and analgesic effects. The underlying mechanisms of the anti-inflammatory pain effects of ligustilide are not completely understood. OBJECTIVE: The aim of this study to investigate whether ligustilide conducts its analgesic effects on the complete Freund's adjuvant (CFA)-induced inflammatory pain through regulating the c-Jun N-terminal kinase (JNK)/c-Jun pathway in the spinal cord. MATERIALS AND METHODS: Paw withdrawal thresholds (PWTs) and paw withdrawal latencies (PWLs) were tested to examine the analgesic effect of ligustilide on CFA-induced inflammatory pain in rats. The change of spinal JNK/c-Jun activation was detected by western blotting after CFA injection with or without consecutive intrathecal ligustilide administration. After SP600125 (JNK inhibitor) was intrathecally injected in CFA rats, PWTs and PWLs were tested to investigate the change of ligustilide's analgesic effect. RESULTS: Repeated intravenous injection of ligustilide could attenuate the pain hypersensitivity induced by CFA. CFA caused increased activation of spinal JNK/c-Jun, which could be inhibited by ligustilide administration. Intrathecal injection of JNK inhibitor inhibited the CFA-induced mechanical hyperalgesia. CONCLUSION: Ligustilide could inhibit the upregulation of spinal p-JNK/p-c-Jun caused by CFA, and the inhibition of JNK/c-Jun activation is closely related to its anti-mechanical hyperalgesia effect in inflammatory pain. SUMMARY: Ligustilide, an active ingredient in a popular traditional Chinese medicine, has effective anti-inflammatory and analgesic effects. Ligustilide inhibits the complete Freund's adjuvant-induced activation of spinal c-Jun N-terminal kinase-(JNK)/c-Jun pathway in rats. The inhibition of JNK/c-Jun activation is closely related to the anti-mechanical hyperalgesia effect of ligustilide. Abbreviations used: CFA: Complete Freund's adjuvant, JNK: c-Jun N-terminal kinase, MAPK: Mitogen-activated protein kinase, PWT: Paw withdrawal threshold, PWL: Paw withdrawal latency.

Koningiopisins A-H, Polyketides with Synergistic Antifungal Activities from the Endophytic Fungus Trichoderma koningiopsis.

Eight new fungal polyketides named koningiopisins A-H (1-8) and four previously known polyketides (9-12) were isolated from the endophytic fungus Trichoderma koningiopsis YIM PH 30002. Their structures were elucidated using extensive spectral data interpretation, and their antifungal and synergistic activities were also evaluated. Koningiopisin C (3) exhibited in vitro antifungal activity against the phytopathogenic fungus Plectosphaerella cucumerina with an MIC of 16 µg/mL. Although the antifungal activities of single compounds were not obvious, a mixture of six compounds (4-9) exhibited potent synergistic antifungal activity against P. cucumerina with an MIC of 16 µg/mL, and the antifungal activity of the mixture of any two compounds with a 1:1 ratio was better than that observed from the individual compound. The synergistic biological activity of the metabolites in YIM PH 30002 demonstrates the significant ecological function of the endophyte for its host plant, and provides additional insight into the search for and development of agents for biological control.

Treatment with ginseng total saponins improves the neurorestoration of rat after traumatic brain injury.

ETHNOPHARMACOLOGICAL RELEVANCE: Ginseng, the root of Panax ginseng C.A. Meyer, is a traditional medicinal herb that has been widely used in Asia for the treatment of many diseases through its effects of reinforcing vitality, strengthening the bodily resistance to pathogenic factors, engendering body liquids and allaying thirst, relieving uneasiness of the body and mind and benefiting intelligence, reducing body weight and prolonging life. Ginsenosides are the most important biologically active substances in ginseng. Many reports have suggested that ginsenosides could exert prominent neuroprotective and neurotrophic effects, promote neural stem/progenitor cell (NSC) proliferation and promote neurite outgrowth and neuronal network formation. The present study aimed to investigate whether treatment with ginsenosides could facilitate NSC proliferation in the hippocampal formation after traumatic brain injury (TBI) and contribute to the recovery of neurological functions including learning and memory. MATERIALS AND METHODS: The modified Feeney׳s method was used to induce a TBI in rats. Ginseng total saponins (GTS) were treated intraperitoneally twice a day for 1 week after the TBI. The neurological functions, morphology of the hippocampus, expression of nerve growth-related factors and number of NSCs in the hippocampal formation ipsilateral to the trauma were determined. RESULTS: We determined 1) GTS (5-80 mg/kg) treatment after a TBI improved the recovery of neurological functions, including learning and memory, and reduced cell loss in the hippocampal area. The effects of GTS at 20, 40, 60, and 80 mg/kg were better than the effects of GTS at 5 and 10 mg/kg. 2) GTS treatment (20 mg/kg) after a TBI increased the expression of NGF, GDNF and NCAM, inhibited the expression of Nogo-A, Nogo-B, TN-C, and increased the number of BrdU/nestin positive NSCs in the hippocampal formation. CONCLUSIONS: GTS treatment in rats after a TBI alleviated the secondary brain injury and ameliorated the neurological functions with an effective dose limit of 5-80 mg/kg. GTS regulated the expression of nerve growth-related factors and improved the proliferation of neural stem/progenitor cells, which might facilitate neural regeneration and tissue repair, and might contribute to the recovery of neurological functions, including learning and memory. These effects of GTS might provide a foundation for the use of ginseng as a medicinal herb to enhance intelligence, reduce the aging process and prolong life in the traditional medicine.

Medelamine C, a new omega-hydroxy alkylamine derivative from endophytic Streptomyces sp. YIM 66142.

A new alkylamine derivative and a common fatty acid were isolated from Streptomyces sp. YIM 66142. On the basis of spectral data, including HRMS, NMR and 2D NMR, their structures were determined as medelamine C (1) and isomyristic acid (2). The omega-hydroxyl group in structure 1 is rare in a natural alkylamine. The possible biosynthetic pathway in the genus Streptomyces from isomyristic acid (2) to medelamines is proposed. Compound 1 showed no obvious cytotoxicity against HL-60, SMMC-7721, A-549, MCF-7, SW480 cell lines. The omega-hydroxyl and the acetyl at NH in compound 1 decreased its cytotoxicity in comparison with that of medelamine.

Blastococcus endophyticus sp. nov., an actinobacterium isolated from Camptotheca acuminata.

A novel endophytic actinobacterium, designated strain YIM 68236(T), was isolated from healthy leaves of Camptotheca acuminata. and characterized by using a polyphasic approach. Cells of this strain occurred singly, in pairs or in tetrads. It grew at 10-45 °C, at pH 5.0-8.0 (optimum pH 7.0) and in the presence of 0-3% (w/v) NaCl. The DNA G+C content was 71.6 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM 68236(T) belongs to the genus Blastococcus. However, it differed from its closest relatives, Blastococcus aggregatus DSM 4725(T), Blastococcus saxobsidens DSM 44509(T) and Blastococcus jejuensis DSM 19597(T) in many phenotypic characteristics. Moreover, the DNA-DNA relatedness values between the novel isolate and the three above-mentioned type strains were 49.0 ± 1.6%, 46.1 ± 3.2% and 39.8 ± 1.5%, respectively. Based on comparative analysis of physiological and chemotaxonomic data, strain YIM 68236(T) represents a novel species of the genus Blastococcus, for which the name Blastococcus endophyticus sp. nov. is proposed. The type strain is YIM 68236(T) ( =CCTCC AA 209045(T) =DSM 45413(T) =KCTC 19998(T)).

Identification of antifungal natural products via Saccharomyces cerevisiae bioassay: insights into macrotetrolide drug spectrum, potency and mode of action.

Since current antifungal drugs have not kept pace with the escalating medical demands of fungal infections, new, effective medications are required. However, antifungal drug discovery is hindered by the evolutionary similarity of mammalian and fungal cells, which results in fungal drug targets having human homologs and drug non-selectivity. The group III hybrid histidine kinases (HHKs) are an attractive drug target since they are conserved in fungi and absent in mammals. We used a Saccharomyces cerevisiae reporter strain that conditionally expresses HHK to establish a high-throughput bioassay to screen microbial extracts natural products for antifungals. We identified macrotetrolides, a group of related ionophores thought to exhibit restricted antifungal activity. In addition to confirming the use of this bioassay for the discovery of antifungal natural products, we demonstrated broader, more potent fungistatic activity of the macrotetrolides against multiple Candida spp., Cryptococcus spp., and Candida albicans in biofilms. Macrotetrolides were also active in an animal model of C. albicans biofilm, but were found to have inconsistent activity against fluconazole-resistant C. albicans, with most isolates resistant to this natural product. The macrotetrolides do not directly target HHKs, but their selective activity against S. cerevisiae grown in galactose (regardless of Drk1 expression) revealed potential new insight into the role of ion transport in the mode of action of these promising antifungal compounds. Thus, this simple, high-throughput bioassay permitted us to screen microbial extracts, identify natural products as antifungal drugs, and expand our understanding of the activity of macrotetrolides.

An endophytic Pseudonocardia species induces the production of artemisinin in Artemisia annua.

Endophytic actinobacteria colonize internal tissues of their host plants and are considered as a rich and reliable source of diverse species and functional microorganisms. In this study, endophytic actinobacterial strain YIM 63111 was isolated from surface-sterilized tissue of the medicinal plant Artemisia annua. We identified strain YIM 63111 as a member of the genus Pseudonocardia. A. annua seedlings grown under both sterile and greenhouse conditions were inoculated with strain YIM 63111. The growth of A. annua seedlings was strongly reduced when YIM 63111 was inoculated at higher concentrations under sterile conditions. However, no growth inhibition was observed when A. annua was grown under greenhouse conditions. Using an enhanced green fluorescent protein (EGFP) expressing YIM 63111 strain, we also observed the endophytic colonization of A. annua seedling using confocal laser-scanning microscopy. The transcription levels of the key genes involved in artemisinin biosynthesis were investigated using real time RT-PCR, revealing that cytochrome P450 monooxygenase (CYP71AV1) and cytochrome P450 oxidoreductase (CPR) expression were up-regulated in A. annua upon inoculation with strain YIM 63111 under certain conditions. The up-regulation of these genes was associated with the increased accumulation of artemisinin. These results suggest that endophytic actinobacteria effectively stimulate certain plant defense responses. Our data also demonstrate the use of Pseudonocardia sp. strain YIM 63111 as a promising means to enhance artemisinin production in plants.

Methods for the study of endophytic microorganisms from traditional Chinese medicine plants.

Plant endophytes are very numerous and widely distributed in nature, their relationships being described as a balanced symbiotic continuum ranging from mutualism through commensalism to parasitism during a long period of coevolution. Traditional Chinese medicines have played a very important role in disease treatment in China and other Asian countries. Investigations show that these medicinal plants harbor endophytes with different kinds of ecological functions, and some of them have potential to produce bioactive small-molecule compounds. This chapter will focus on the selective isolation methods, the diversity of some endophytes (actinobacteria and fungi) isolated from Traditional Chinese Medicine (TCM) plants, and the bioactive compounds from selected endophytic actinobacteria reported in the past 3 years.

Abundant and diverse endophytic actinobacteria associated with medicinal plant Maytenus austroyunnanensis in Xishuangbanna tropical rainforest revealed by culture-dependent and culture-independent methods.

Endophytes are now considered as an important component of biodiversity. However, the diversity of endophytic actinobacteria associated with tropical rainforest native medicinal plants is essentially unknown. In this study, the diversity of endophytic actinobacteria residing in root, stem and leaf tissues of medicinal plant Maytenus austroyunnanensis collected from tropical rainforest in Xishuangbanna, China was investigated with a combination of cultivation and culture-independent analysis on the basis of 16S rRNA gene sequencing. By using different selective isolation media and methods, a total of 312 actinobacteria were obtained, and they were affiliated with the order Actinomycetales (distributed into 21 genera). Based on a protocol for endophytes enrichment, three 16S rRNA gene clone libraries were constructed and 84 distinct operational taxonomic units were identified and they distributed among the orders Actinomycetales and Acidimicrobiales, including eight suborders and at least 38 genera with a number of rare actinobacteria genera. Phylogenetic analysis showed that 32% of the clones in the libraries had lower than 97% similarities with related type strains. Interestingly, six genera from the order Actinomycetales and uncultured clones from Acidimicrobiales have not, to our knowledge, been previously reported as endophytes. Our study confirms abundant endophytic actinobacterial consortium in tropical rainforest native plant and suggests that this special habitat represents an underexplored reservoir of diverse and novel actinobacteria of potential interest for bioactive compounds discovery.

Rhodococcus artemisiae sp. nov., an endophytic actinobacterium isolated from the pharmaceutical plant Artemisia annua L.

A Gram-positive, non-motile actinobacterium, designated YIM 65754T, was isolated from the stem of Artemisia annua L., collected from Yunnan province, south-west China. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain YIM 65754T comprised an evolutionary lineage within the genus Rhodococcus. The isolate clustered with Rhodococcus pyridinivorans PDB9T, Rhodococcus gordoniae W 4937T and Rhodococcus rhodochrous DSM 43241T, with which it shared 98.4, 97.9 and 97.8 % 16S rRNA gene sequence similarities, respectively. However, DNA-DNA relatedness demonstrated that strain YIM 65754T was distinct from its closest phylogenetic neighbours. The cell-wall peptidoglycan contained meso-diaminopimelic acid, arabinose, galactose, mannose and glucose (cell-wall chemotype IV). The major menaquinone was MK-8(H2) and the predominant fatty acids were C16:0 (27.83 %), iso-C15:0 2-OH and/or C16:1ω7c (20.21 %) and 10-methyl C18:0 (17.50 %). The DNA G+C content was 66.2 mol%. On the basis of phenotypic, chemotaxonomic and phylogenetic evidence, the isolate represents a novel species of the genus Rhodococcus, for which the name Rhodococcus artemisiae sp. nov. is proposed; the type strain is YIM 65754T (=CCTCC AA 209042T=DSM 45380T).

Promicromonospora xylanilytica sp. nov., an endophytic actinomycete isolated from surface-sterilized leaves of the medicinal plant Maytenus austroyunnanensis.

A novel xylan-degrading actinomycete, strain YIM 61515(T), was isolated from surface-sterilized leaves of the medicinal plant Maytenus austroyunnanensis. Cells were Gram-positive and non-spore-forming, produced primary branches and formed white to yellowish white colonies on the media tested. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM 61515(T) was most similar to Promicromonospora aerolata V54A(T) and Promicromonospora vindobonensis V45(T) (99.4 and 99.1% sequence similarity, respectively). The isolate formed a separate lineage in a cluster containing P. aerolata V54A(T). 16S rRNA gene sequence similarity between the isolate and other members of the genus Promicromonospora ranged from 96.3 to 98.4%. Chemotaxonomic data, including major menaquinones, fatty acid compositions and polar lipid profiles, supported the placement of strain YIM 61515(T) in the genus Promicromonospora. DNA-DNA relatedness, physiological and biochemical data showed that strain YIM 61515(T) could be distinguished from members of all known species of the genus Promicromonospora and therefore represented a novel species. The name Promicromonospora xylanilytica sp. nov. is proposed, with YIM 61515(T) (=DSM 21603(T)=CCTCC AA 208046(T)) as type strain.

[Screening cold-acclimation differential expression candidate genes in the brain of common carp (Cyprinus carpio)].

In this study, 26 candidate genes were quantified and normalized in the brain cDNA of common carp (Cyprinus carpio) at 23°C and 6°C using double-standard curve method of real-time quantitative PCR. The results showed that five candidates up-regulated in the samples at 6°C (P<0.01) and quantified 2.11, 13.9, 2.52, 7.38, and 1.83 times more than in the samples at 23°C, respectively. Gene function searching indicated that the protein products of these five candidates were elongation of very long chain fatty acids protein, Acyl-CoA desaturase, Transcription initiation factor IIB, Myo-inositol- 1-phosphate synthase, and Blood-brain barrier HT7 antigen individually. Moreover, seven down-regulated candidates were also identified in the same samples at 6°C (P>0.05), and their expression levels were decreased by 21.8%, 25.9%, 16.6%, 23.7%, 15.8%, 16.3%, and 42.5%, respectively, in comparison with the samples at 23°C. These seven down-regulated candidates mainly participated in the inhibition of glycolysis, improvement of cell apoptosis, and intervention of synapse remodeling based on the results of function searching. The five cold-induced genes identified in this study will be used as important elements for fish with cold sensitive through transgenic technology in future.

Isolation, diversity, and antimicrobial activity of rare actinobacteria from medicinal plants of tropical rain forests in Xishuangbanna, China.

Endophytic actinobacteria are relatively unexplored as potential sources of novel species and novel natural products for medical and commercial exploitation. Xishuangbanna is recognized throughout the world for its diverse flora, especially the rain forest plants, many of which have indigenous pharmaceutical histories. However, little is known about the endophytic actinobacteria of this tropical area. In this work, we studied the diversity of actinobacteria isolated from medicinal plants collected from tropical rain forests in Xishuangbanna. By the use of different selective isolation media and methods, a total of 2,174 actinobacteria were isolated. Forty-six isolates were selected on the basis of their morphologies on different media and were further characterized by 16S rRNA gene sequencing. The results showed an unexpected level of diversity, with 32 different genera. To our knowledge, this is the first report describing the isolation of Saccharopolyspora, Dietzia, Blastococcus, Dactylosporangium, Promicromonospora, Oerskovia, Actinocorallia, and Jiangella species from endophytic environments. At least 19 isolates are considered novel taxa by our current research. In addition, all 46 isolates were tested for antimicrobial activity and were screened for the presence of genes encoding polyketide synthetases and nonribosomal peptide synthetases. The results confirm that the medicinal plants of Xishuangbanna represent an extremely rich reservoir for the isolation of a significant diversity of actinobacteria, including novel species, that are potential sources for the discovery of biologically active compounds.

Glycomyces scopariae sp. nov. and Glycomyces mayteni sp. nov., isolated from medicinal plants in China.

Two actinomycete strains, designated YIM 56256(T) and YIM 61331(T), were isolated from the roots of Scoparia dulcis and Maytenus austroyunnanensis, two Chinese medicinal plants, and their taxonomic status was established based on a polyphasic investigation. The organisms were found to have chemical and morphological markers typical of members of the genus Glycomyces. 16S rRNA gene sequence analysis showed that they were closely related to each other and to Glycomyces sambucus E71(T). A battery of physiological characteristics and levels of DNA-DNA relatedness indicated that strains YIM 56256(T) and YIM 61331(T) represent two novel species, clearly different from the related known Glycomyces species. On the basis of the data presented, it is evident that each of these strains represents a novel species of the genus Glycomyces, for which the names Glycomyces scopariae sp. nov. (type strain YIM 56256(T) =KCTC 19158(T) =DSM 44968(T)) and Glycomyces mayteni sp. nov. (type strain YIM 61331(T) =KCTC 19527(T) =CCTCC AA 208004(T)) are proposed.

Pseudonocardia endophytica sp. nov., isolated from the pharmaceutical plant Lobelia clavata.

An endophytic actinomycete strain, designated YIM 56035(T), was isolated from the inner tissue of the traditional Chinese medicinal plant Lobelia clavata. The strain stained Gram-positive, was aerobic and exhibited branching, white aerial mycelium and yellowish-brown substrate mycelium. The strain formed spore chains on aerial hyphae. The G+C content of the genomic DNA was 70.3 mol%. On the basis of morphological, physiological, chemotaxonomic and phylogenetic characteristics, strain YIM 56035(T) was assigned to the genus Pseudonocardia. Analysis of 16S rRNA gene sequences showed 98.5, 97.3, 97.3 and 97.1 % similarity to the closely related type strains Pseudonocardia kongjuensis LM 157(T), Pseudonocardia autotrophica IMSNU 20050(T), Pseudonocardia ammonioxydans H9(T) and Pseudonocardia compacta IMSNU 20111(T), respectively. The results of DNA-DNA hybridizations and comparison of some phenotypic characteristics revealed that the strain represents a novel species of the genus Pseudonocardia. The name Pseudonocardia endophytica sp. nov. is proposed, with the type strain YIM 56035(T) (=DSM 44969(T) =CCTCC AA 206026(T) =KCTC 19150(T)).