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In this study, we developed a repeatable in vitro micropropagation protocol for the medicinal plant Asparagus cochinchinensis, based on indirect organogenesis using leaf segments cut from seedlings of in vitro-germinated seeds. We obtained 85% callus induction from the leaf segments within 4–5 weeks when grown on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BAP, 1.0 mg/L) and 1-naphthaleneacetic acid (NAA, 0.5 mg/L). We found that MS media supplemented with Kn (1.0 mg/L) in combination with NAA (1.0 mg/L) and BAP (1.0 mg/L) combined with NAA (0.5 mg/L) were the most effective in promoting shoot regeneration, yielding plantlets with 6.72 and 6.48 shoots per culture, respectively. When cultured on PGR-free half-strength MS medium, regenerated plants developed root systems with an average 11.0 roots per shoot cluster and an average length of 36.14 mm at 9 weeks. During acclimatization, regenerated plantlets showed 96.4% survival and exhibited normal growth characteristics and morphology. We also made an attempt to directly regenerate A. cochinchinensis from shoot apices but it was futile. Histological analyses revealed the presence of crystal idioblasts in young leaves from the early stages of leaf differentiation. The leaf-based plant regeneration technique developed herein could be employed for large-scale propagation of the plants over a short time period, thereby substantially contributing to the germplasm preservation and rapid propagation of A. cochinchinensis. A repeatable in vitro micropropagation protocol for Asparagus cochinchinensis was developed based on indirect organogenesis. Histological analysis revealed crystal idioblasts for the first time in leaf primordia of this species.

This present study evaluated the effects of processed P. multiflorum on osteogenesis using Sarcoma osteogenic (SaOS-2) cell lines and osteoclastogenesis of bone marrow-derived macrophage cells (BMM) and to elucidate differences in effect on the expression of bone-related proteins between commercially sold P. multiflorum and patented, in vitro-propagated Korea Institute of Oriental Medicine (KIOM) P. multiflorum. Raw P. multiflorum and P. multiflorum that were stir-baked and steamed in black bean juice were compared, and western blotting analysis was performed to investigate the expression of bone remodeling-related proteins in SaOS-2 cells. In the cells treated with P. multiflorum steamed in black bean juice, the expression of RANKL was decreased, whereas that of osteoprotegerin, alkaline phosphatase, Runx2, and osterix was increased. Owing to these results, we conclude that processed P. multiflorum can be used as an alternative treatment for bone diseases such as osteoporosis, osteopenia, periodontitis, and Paget’s disease.

Background: The genus Prunus (Family Rosaceae) comprises over 400 plant species and exhibits vast biodiversity worldwide. Due to its wide distribution, its taxonomic classification is important. Anatomical characters are conserved and stable and thus can be used as an important tool in plant taxonomic characterization. Thus, this study aimed at examining and documenting P. africana leaf, stem, and seed anatomy using micrographs and photographs for possible use in identification, quality control, and phylogenetic studies of the species.Methods: P. africana leaves, stems, and seeds were fixed, dehydrated in ascending ethanol series (50–100 %), embedded in Technovit resin, and sectioned using a microtome for mounting histological slides for anatomical observation under a microscope and subsequent description.Results: The anatomical sections of a young stem revealed a cortex consisting of isodiametric parenchyma cells, druse crystals, primary vascular bundles, and pith. The mature stem bark consisted majorly of rhytidome with periderm densely arranged in multiple layers, a cluster of stone cells, and sclerenchyma. The sections of the leaf were hypostomatic with stomata size ranging between 18.90– (22.34)–26.90 × 15.41– (18.40)–21.22 μm. The leaf sections showed the presence of characteristic druse crystals, vascular bundles, and mesophyll layers. The pericarp showed the presence of epicarp, mesocarp, and endocarp with a thickness of approximately 350–400, 300–350, and 30–50 μm, respectively and a seed testa with a thickness of approximately 50–60 μm. Conclusion: The characteristic morphological and anatomical features observed in P. africana leaves, stems, and seeds in this study could provide useful data in taxonomical identification of this species.

Trichosanthes kirilowii Maxim. is a vital traditional herbal medicinal plant found in northeastern Asia. Its roots, fruits, and seeds are used as food and medicine. Roots harvested for medicinal use take over 3 yr to mature when the plant is grown in a traditional way through cultivation in the field. This coupled with uncertainty in identification of the plant when collected from the wild calls for a standard in vitro propagation system to meet the increasing demand for it. The purpose of this study was to develop a standard protocol for the in vitro micropropagation of T. kirilowii. Ten different media supplemented with different concentrations of plant growth regulators were evaluated. At 5 wk, De Greef and Jacobs medium supplemented with 0.1 mg L−1 kinetin led to optimal shoot growth, while the same medium supplemented with 0.5 mg L−1 indole 3-butyric acid induced optimal root growth, also at 5 wk. The micropropagated plants that were acclimatized for 8 wk in the greenhouse produced mature root tubers after planted in the field for 3 mo. Therefore, these findings provide a basis for future large-scale in vitro propagation of T. kirilowii.

Chronic inflammation is one of the causes of a number of non-infectious diseases in the world. Over the years, Tamarindus indica has played fundamental roles in traditional medicine as an anti-inflammatory and analgesic drug. It is a commercialized biocompatible medicinal plant species with a wide range of therapeutic window and with suggested LD50 greater than 5000 mg kg−1 body weight when administered to the Wistar rats. This review examined the anti-inflammatory and analgesic potential and mechanism of various extracts from T. indica pulp, leaves, seeds, stem bark, and roots. The preclinical studies provided strong pharmacological evidence for the anti-inflammatory and analgesic activities of the different parts of T. indica and this may be attributed to the various bioactive compounds in it including alkaloids, flavonoids, tannins, phenols, saponins, and steroids. The anti-inflammatory and analgesic effects of the extracts from the different parts of T. indica may be due to its ability to inhibit a number of biological processes including cyclooxygenase-2 (COX-2) expression, inducible nitric oxide synthase (iNOS), 5-lipoxygenase biosynthesis, and tumor necrosis factor-α. The analgesic activity of T. indica may also be through the activation of the opioidergic mechanism at both the peripheral and central levels. Although further pre-clinical studies still need to be conducted, these results demonstrated that T. indica has potent anti-inflammatory and analgesic activities and hence provides justification for its use in traditional medicine to treat body pain and other inflammatory related diseases including arthritis and offers a basis for future clinical studies and possible drug development.

Aspilia africana has been used for generations to treat many diseases in Africa. Its biological activities, including antioxidant and anti-inflammatory potential, are attributed to a number of secondary metabolites, including alkaloids and polyphenolics. The antioxidant activities of A. africana callus (CA), juvenile in vitro leaf (IL) and root (IR), ex vitro root (SR) and leaf (SL), and wild leaf (WL) dried samples were assessed based on their diphenylpicrylhydrazyl (DPPH) free radical scavenging abilities. The total phenolic and flavonoid content of different plant samples was compared. Further, high-pressure liquid chromatography (HPLC) was used to quantitatively determine chlorogenic acid content in the A. africana plant samples. Fourier transform near-infrared (FT-NIR) analysis was also carried out to compare the antioxidant phytochemical content in the A. africana plant tissues. Among the samples, IR, with the highest total phenolic content (167.84 ± 1.057 mg GAE/g), total flavonoid content (135.06 ± 0.786 mg RUE/g), and chlorogenic acid (5.23 ± 0.298 mg/g) content, had the most potent antioxidant activity (IC50 = 27.25 ± 5.028 μg/mL), followed by WL. The lowest polyphenolic content and antioxidant activity were observed in SR. The antioxidant activities of A. africana tissues were positively correlated with the total phenolic and flavonoid content in the samples. The differences in antioxidant activities of A. africana tissues could be attributed to the difference in their polyphenolic content. Our study reports, for the first time, the antioxidant activities of A. africana callus and roots (in vitro and ex vitro). The A. africana samples IR, CA, and WL could be valuable natural sources of antioxidants that could be further exploited for the development of useful pharmaceutical products.

Prunus africana is an endangered medicinal plant and hence new propagation methods are urgently required to increase its populations. Unfortunately, propagation through seeds is challenging due to its long flowering cycle and recalcitrant seeds. We developed a protocol for micropropagation using nodal segment explants. A woody plant medium supplemented with vitamins, 15 g L−1 sucrose, and 1.0 mg L−1 6-benzylaminopurine (BAP) supported the optimum rate (100%) of axillary shoot initiation. Supplementation with 15 g L−1 sucrose and 1.5 mg L−1 indole-3-acetic acid (IAA) provided the optimum rate (75%) of root initiation. Rooted plantlets were successfully planted in sterilized horticultural soil containing perlite (2:1 v/v) and the survival rate was 98% following acclimatization. The photosynthetic rate assessed using FlourPen FP110 series showed that the ratio of variable fluorescence to maximum fluorescence mean value for in vitro regenerated P. africana (0.830 ± 0.0008) was similar to that of the maternal P. africana plant (0.825 ± 0.005), indicating similarity in their photosynthetic performance; a pivotal process for growth and development. The Fourier transform near-IR (FT-NIR) spectrometer analysis of the in vitro regenerated and the maternal P. africana plant samples exhibited homogeneity in the absorbance peaks at 8,273, 6,344, and 4,938–4,500 cm−1 associated with lipids, starch, and proteins. The genetic fidelity of regenerated plants was confirmed using the randomly amplified polymorphic DNA (RAPD) technique. Our protocol is suitable for use in large-scale P. africana to meet the increasing demands for it in the global market.

Polygonum multiflorum Thunb. is a perennial plant that belongs to Polygonaceae. Root tissues are the main plant parts used as medicinal herbs in Korean oriental medicine. The P. multiflorum tuber is well known for its medicinal properties in Korean oriental medicine, and it contains a number of useful substances (secondary metabolites of emodin, 2,3,5,4′-tetrahydroxystilbene-2-O-β-d-glucoside (TSG), etc.) that are increasing in demand, as several studies show that they have beneficial effects on the human body. In this study, the production volumes and useful material content differences between cultured P. multiflorum seedlings (culture seedlings: CSs), which had been grown using a tissue culture technique under optimized conditions, and existing varieties in circulation (seed seedlings: SSs) were determined using a long-term field test. The growth characteristics of the underground parts were investigated by harvesting the tuberous roots (medicinal parts) after 1 year, and the results showed that the fresh and dry weights of the CS tubers were higher than those of the SS tubers. However, the SS rootlets had higher fresh and dry weights than the CS rootlets. A liquid chromatography-mass spectrometry component analysis of the P. multiflorum tubers and a Fourier transform near-infrared spectrophotometer analysis of the roots were undertaken. The results showed that the levels of TSG, which is a medicinal substance produced by P. multiflorum, were higher in the CSs than in the SSs, but the differences were not significant. The CS results from this study will inform future studies on the mass production of P. multiflorum in the field because the medicinal area was greater in CSs than in SSs.

Osteoporosis is one of the main health problems in the world today characterized by low bone mass and deterioration in bone microarchitecture. In recent years, the use of natural products approach to treat it has been in the increase. In this study, in vitro antiosteoporosis activity and hepatotoxicity of P. jamasakura bark extracts were evaluated. Methods. Mouse bone marrow macrophage (BMM) cells were incubated with tartrate-resistant acid phosphate (TRAP) buffers and p-nitrophenyl phosphate and cultured with different P. jamasakura bark extracts at concentrations of 0, 6.25, 12.5, 25, and 50 μg/ml in the presence of the receptor activator of nuclear factor kappa-Β ligand (RANKL) for 6 days. The osteoclast TRAP activity and cell viability were measured. Nitric oxide (NO) assay was conducted using murine macrophage-like RAW 264.7 cells treated with P. jamasakura ethanolic and methanolic bark extracts at concentrations of 0, 6.25, 12.5, 25, 50, 100, and 200 μg/ml. For hepatotoxicity assessment, zebrafish larvae were exposed to P. jamasakura bark extracts, 0.05% dimethyl sulfoxide as a negative control, and 5 μM tamoxifen as a positive control. The surviving larvae were anesthetized and assessed for hepatocyte apoptosis. Results. TRAP activity was significantly inhibited (p P. jamasakura extracts compared to the control treatment. At 50 μg/ml, both ethanolic and methanolic extracts of P. jamasakura exhibited significant (p P. jamasakura ethanolic and methanolic extracts had significant inhibitory (p μg/ml and exhibited significant (p p Conclusion. The TRAP activity of P. jamasakura bark gives a foundation for further studies to enhance future development of antiosteoporosis drug.

Rehmannia glutinosa (Gaertn.) DC is a perennial plant belonging to the family Scropulariidae. The root of R. glutinosa is used in oriental medicine and mainly grown using rootstock rather than seed cultivation, which gives rise to several problems including root rot, and results in a low productivity and poor quality. To solve the challenges involved in R. glutinosa seed cultivation, our team previously used the formative features and genetic analysis of R. glutinosa to determine the optimal in vitro tissue culture conditions for producing sterile culture seedlings and rootstocks of R. glutinosa. The aim of the present study was to identify differences between R. glutinosa standard rootstock seedlings (SR), R. glutinosa culture rootstock seedlings (CR), and culture seedlings (CS) under field conditions. The reproductive characteristics of the aerial part were more robust while the area and length of leaves were smaller for SR than those for CR and CS. The characteristic that differed the most in SR was flowering, which did not occur in CR and CS. In addition, the fresh and dry weights of the subterranean parts of CR and CS were two-fold greater than those of SR. Fourier transform near-infrared (FT-NIR) analysis showed only slight differences between the chemical constituents of SR and its culture products, which was confirmed by measuring the content of catalpol, an indexing substance. Catalpol had a reduced content in the culture products compared to SR. However, this difference was not significant. Our findings will be useful for the identification of the best seedling type of R. glutinosa to enable its mass production.

Warburgia ugandensis Sprague (Family Canellacea) commonly known as Ugandan greenheart or pepper bark tree, is a highly valued medicinal plant in traditional medicine with a broad spectrum of antimicrobial activity whose parts especially the leaves and stem bark have for long been used in the treatment and management of many diseases and health conditions such as stomachache, cough, toothache, fever, malaria, oral thrush, measles and diarrhea in African communities where the plant occurs. This review focused on the phytochemical contents, medicinal uses and antimicrobial activities of W. ugandensis based on published peer reviewed articles. This review established that the high therapeutic value of W. ugandensis is attributed to the abundance of drimane sesquiterpenes in its stem bark and leaves. These chemicals have also made the plant to have potent antibacterial and antifungal activities. However, more pre-clinical and clinical trials need to be done to further validate the traditional medicine applications of W. ugandensis for possible drug discovery. Due to its high demand, W. ugandensis has been over exploited and hence its population is in drastic decline. Consequently, there is need for development of advanced and more rapid propagation techniques to increase its population and distribution in its natural environment to meet the ever-increasing demand. Key words: Antimicrobial, Canellaceae, medicinal uses, phytochemicals, Warburgia ugandensis.

Rehmannia glutinosa is an important medicinal crop that has been widely used globally to treat various ailments such as hematinic deficiencies, diabetes, and adrenal disorders. To provide a sustainable supply to meet large demands, alternative cultivation methods, apart from conventional ones, are indispensable. This study developed an optimized condition for R. glutinosa adventitious root culture and a scale-up production system using a bioreactor. A woody plant medium enriched with vitamins, 30 g/L sucrose, and 2.0 mg/L 1-naphthaleneacetic acid (NAA) supported the optimum growth rate of adventitious roots (growth index of 25.67). The application of methyl jasmonate (MeJa) at a concentration of 200 μM showed the best enhancement of acteoside biosynthesis, with an approximately 2.53-fold increase (14.37 mg/g dry weight [DW]) compared with the control (5.67 mg/g DW). However, all treatments with MeJa and salicylic acid reduced the biomass of R. glutinosa adventitious roots compared with control. To validate the optimized protocol, the culture system was scaled up by comparing shake flask cultures with 2- and 20-L balloon type (air lift) bubble bioreactors (BTBB) along with elicitation using 200 μM MeJa. The 20-L BTBB cultures produced good-quality R. glutinosa roots with higher biomass (15.94 g DW), acteoside content (22.13 mg/g DW), and antioxidant activity (84.22 %) than shake flask cultures. In addition, fourier transform near-IR (FT-NIR) analysis showed no significant difference in the chemical composition of the in vitro adventitious roots and those that were commercially available. Thus, these findings will provide a basis for the development of large-scale R. glutinosa adventitious root production to meet the demands of pharmaceutical and oriental medicine industries without affecting its availability in nature.

Prostate cancer remains one of the major causes of death worldwide. In view of the limited treatment options for patients with prostate cancer, preventive and treatment approaches based on natural compounds can play an integral role in tackling this disease. Recent evidence supports the beneficial effects of plant-derived phytochemicals as chemopreventive and chemotherapeutic agents for various cancers, including prostate cancer. Prunus africana has been used for generations in African traditional medicine to treat prostate cancer. This review examined the potential roles of the phytochemicals from P. africana, an endangered, sub-Saharan Africa plant in the chemoprevention and chemotherapy of prostate cancer. In vitro and in vivo studies have provided strong pharmacological evidence for antiprostate cancer activities of P. africana-derived phytochemicals. Through synergistic interactions between different effective phytochemicals, P. africana extracts have been shown to exhibit very strong antiandrogenic and antiangiogenic activities and have the ability to kill tumor cells via apoptotic pathways, prevent the proliferation of prostate cancer cells, and alter the signaling pathways required for the maintenance of prostate cancer cells. However, further preclinical and clinical studies ought to be done to advance and eventually use these promising phytochemicals for the prevention and chemotherapy of human prostate cancer.

Wounds remain one of the major causes of death worldwide. Over the years medicinal plants and natural compounds have played an integral role in wound treatment. Aspilia africana (Pers.) C. D. Adams which is classified among substances with low toxicity has been used for generations in African traditional medicine to treat wounds, including stopping bleeding even from severed arteries. This review examined the potential of the extracts and phytochemicals from A. africana, a common herbaceous flowering plant which is native to Africa in wound healing. In vitro and in vivo studies have provided strong pharmacological evidences for wound healing effects of A. africana-derived extracts and phytochemicals. Singly or in synergy, the different bioactive phytochemicals including alkaloids, saponins, tannins, flavonoids, phenols, terpenoids, β-caryophyllene, germacrene D, α-pinene, carene, phytol, and linolenic acid in A. africana have been observed to exhibit a very strong anti-inflammatory, antimicrobial, and antioxidant activities which are important processes in wound healing. Indeed, A. africana wound healing ability is furthermore due to the fact that it can effectively reduce wound bleeding, hasten wound contraction, increase the concentration of basic fibroblast growth factor (BFGF) and platelet derived growth factor, and stimulate the haematological parameters, including white and red blood cells, all of which are vital components for the wound healing process. Therefore, these facts may justify why A. africana is used to treat wounds in ethnomedicine.

The yeast strain Metschnikowia persimmonesis Kang and Choi et al., sp. nov. [type strain KIOM_G15050 = Korean Collection for Type Cultures (KCTC) 12991BP] was isolated from the stalk of native persimmon cultivars (Diospyros kaki Thumb) obtained from different regions of South Korea and was characterized phenotypically, genetically, and physiologically. The isolate grew between 4 and 40 °C (optimum temperature: 24–28 °C), pH 3–8 (pH optimum = 6.0), and in 0–4% NaCl solution (with optimal growth in absence of NaCl). It also exhibited strong antibiotic and antimicrobial activities. Morphologically, cells were characterized by the presence of long, needle-shaped ascospores. Based on 18S ribosomal DNA gene sequence analysis, the new species was found to belong to the genus Metschnikowia as a sister clade of Metschnikowia fructicola. We therefore conclude that this yeast isolate from D. kaki is a new member of the genus Metschnikowia and propose the name M. persimmonesis sp. nov. This strain has been deposited in the KCTC for future reference. This discovery provides a basis for future research on M. persimmonesis sp. nov., including its possible contribution to the medicinal properties of the host persimmon plant.

Background Natural products play a significant role in human health in relation to the prevention and treatment of inflammatory conditions. One of the plants with great medicinal potentials is Diospyros kaki which is mainly cultivated in Asian countries including Korea, Japan, and China. Astringent D. kaki is a wild species with an astringent taste until they are Ripened. kaki calyx is a traditional Korean medicine (TKM) made from the stalks of astringent D. kaki and is used in treating bed-wetting, vomiting, and hiccupping. The present study was designed to investigate the potential anti-inflammatory activities of astringent D. kaki stalks based on cultivar types and stages of maturity. Methods The anti-inflammatory effects of the stalk extracts of local astringent D. kaki cultivar species were evaluated on RAW 264.7 cells. Cell viability was measured using a Cell Counting Kit-8 (CCK8) method. The anti-inflammatory effects were determined by measuring the nitric oxide (NO) concentration of the supernatant. Cellular signaling pathways were determined by quantitative polymerase chain reactions of inducible nitric oxide synthase (iNOS). Protein expression of iNOS and phospho-p65 was determined using western blot, and the nuclear localization of p65 was determined using confocal imaging in RAW 264.7 cells. Results We found that the stage 1 (8–9 month) samples all showed a high percentage of tannic acid content and Gojongsi (Hamyang) stalks had the highest content. The stage 1 samples also showed the highest inhibition of NO production. Decreases in the expression of iNOS and phosphorylated p65, and in the nuclear localization of p65, were dose-dependent. All the extracts were nontoxic under 100 μg/ml concentration. Conclusion This study provides insight into the changes in tannic acid content in astringent D. kaki and their anti-inflammatory effects, in relation to their stage of maturity. These results are expected to be useful in the verification of the efficacy of oriental medicine and the timing of proper harvest for medical use.

Medicinal plants have been an integral part of the healthcare system for centuries. Prunus africana, an evergreen plant of the family Rosaceae in sub-Saharan Africa, has been used to treat numerous diseases and disorders since time immemorial. Over the past decades, several ethnomedicinal surveys were conducted in many communities across Africa which validated the use of P. africana in traditional medicine. The increased interest in P. africana across the globe has led to several pharmacological studies that have further validated the continued use of this plant in traditional medicine. The aim of this study was therefore to review the various evidence based ethnopharmacological studies on the use of P. africana in traditional medicine in treatment and management of benign prostatic hyperplasia, prostate cancer, diabetes, malaria, chest pain, gastrointestinal conditions, wound healing, and skin infections based on peer-reviewed electronic databases. Scientific studies attributed the effectiveness of P. africana in the treatment and management of these diseases to the class of bioactive compounds contained in it, including tannins, saponins, alkaloids, flavonoids, terpenoids, phytosterols, and fatty acids. The anti-inflammatory, antimicrobial, antiandrogenic, antiangiogenic, antioxidant, anti-dipeptidyl peptidase-4, analgesic, and astringent effects of these phytochemicals makes P. africana very effective in the treatment and management of the above diseases. This review therefore provides the foundation of further preclinical and clinical studies on P. africana to enhance future production of high quality, safe, and efficacious medicines for treatment of benign prostatic hyperplasia, prostate cancer, diabetes, malaria, chest pain, gastrointestinal conditions, wound healing, and skin infections.