Anoectochilus Formosanus Research Articles

Protective Effect of Anoectochilus formosanus Polysaccharide against Cyclophosphamide-Induced Immunosuppression in BALB/c Mice.

In this study, Anoectochilus formosanus polysaccharide (AFP) was acquired a via water extraction and alcohol precipitation method. The immunoregulatory activity of AFP was first evaluated on cyclophosphamide (Cy)-treated mice. Galacturonic acid, glucose and galactose were confirmed to be the main components of AFP. AFP demonstrated the ability to stimulate the production of TNF-α and IL-6 in RAW 264.7 macrophages. Not surprisingly, the activation of the NF-κB signaling pathway by AFP was validated via Western blot analysis. Furthermore, AFP could alleviate Cy-induced immunosuppression, and significantly enhance the immunity of mice via increasing the thymus index and body weight, stimulating the production of cytokines (IgA, IgG, SIgA, IL-2, IL-6 and IFN-γ). The improvement in the intestinal morphology of immunosuppressed mice showed that AFP could alleviate Cy-induced immune toxicity. These results have raised the possibility that AFP may act as a natural immunomodulator. Overall, the study of AFP was innovative and of great significance for AFP's further application and utilization.

Polysaccharide and ethanol extracts of Anoectochilus formosanus Hayata: Antioxidant, wound-healing, antibacterial, and cytotoxic activities

IntroductionPolysaccharide and alcohol extracts of Anoectochilus formosanus Hayata have attracted great attention as they exhibit noteworthy properties such as prebiotic and anti-hyperglycemic effects. However, the antioxidant and wound-healing activities of the polysaccharide extract as well as the antibacterial and cytotoxic effects of the ethanol extracts have not been thoroughly uncovered. Therefore, our study investigated these bioactivities of the two extracts prepared from Anoectochilus formosanus to broaden understandings of medical benefits of the plant. MethodsThe monosaccharide composition was analyzed by HPAEC-PAD. The antioxidant and wound-healing activities of the polysaccharide extract were evaluated by ABTS and scratch assays, respectively. The broth dilution method was used to determine the antibacterial ability of the ethanol extract. Additionally, the cytotoxic and mechanistic effects of this extract against hepatocellular carcinoma HUH-7 cells was assessed by MTT assay, qRT-PCR and Western blotting methods. ResultsThe polysaccharide extract possessed an effective free radical scavenging ability in an ABTS assay (IC50 = 44.92 μg/ml). The extract also ameliorated wound recovery in a fibroblast scratch assay. Meanwhile, the ethanol extract was able to inhibit the growth of Staphylococcus aureus (MIC = 2500 μg/ml), Bacillus cereus (MIC = 2500 μg/ml), Escherichia coli (MIC = 2500 μg/ml), and Pseudomonas aeruginosa (MIC = 1250 μg/ml). Additionally, it repressed the viability of HUH-7 cells (IC50 = 53.44 μg/ml), possibly through upregulating the expression of caspase 3 (CASP3), CASP8, and CASP9 at both mRNA and protein levels. ConclusionThe polysaccharide extract of A. formosanus exhibited the antioxidant and wound-healing properties whereas the ethanol extract showed the antibacterial activity and cytotoxicity against HUH-7 cells. These findings specify notable biological effects of the two extracts which could be of potential use in human healthcare.

In silico comparative analysis of the complete chloroplast genome sequences in different jewel orchid species

Abstract. Nguyen MP, Trinh TH, Ngo TKA, Widiarsih S, Ho VT. 2023. In silico comparative analysis of the complete chloroplast genome sequences in different jewel orchid species. Nusantara Bioscience 15: 12-21. Jewel orchid is the common name of several orchid species which can be alike in morphological characteristics but variable in medicinal properties. As these plants are utilized to treat several diseases, their natural existence in the wild habitat is rapidly diminished. Therefore, a better understanding of the genetic information of this plant for better genetic conservation and development of these plants is necessary. In this study, a total of 18 published chloroplast genomes of 18 jewel orchid species determined by the next-generation sequencing method were retrieved from NCBI GenBank and targeted for genomic characterization and phylogenetic analyses. Different bioinformatics tools were utilized to characterize these genomes’ genomic structure, repetitive sequences, interspecific variation, divergence, and phylogenetic relationships. The obtained data revealed that the chloroplast genomes of different jewel orchid species varied in length between 151,414 (Anoectochilus formosanus MN880624.1) and 154,375 (Goodyera biflora OM314910.1). Each species contains 34-87 SSR loci which could be useful as molecular markers for further genetic diversity study of this plant. Structural variations in the expansion and contraction of inverted repeat regions were also considered. Phylogenetic analysis identified a close relationship among species belonging to the Goodyera genus, and this genus is distinctive from other genera such as Anoectochilus, Cystorchis, Dossinia, Ludisia, and Macodes. The obtained results show a high potential of deeper characterizing the chloroplast genome of jewel orchids for species classification, identification, molecular breeding, and evolutionary exploration of these important herbal plants.

Unravelling the therapeutic potential of orchid plant against cancer

Cancer is one of the deadly diseases all over the world which is defined as unregulated growth and division of the cells. Natural remedies, particularly derived from plants, were used to treat a range of illnesses for thousands of years, including cancer. Several orchid species have been reported as a cure to treat various types of cancers. Orchids are huge family of flower bearing plants. They are traditionally used in folk medicine for the cure of various infectious diseases and tumour. Many of bioactive compounds isolated from orchid plants such as Dendrobium longicornu Lindl., Dendrobium transparens, Rhyncostylish retusa , Vanda cristata, Anoectochilus formosanus etc. possess anticancer activities via modifying the biotransformation of potential carcinogens by xenobiotic-metabolizing enzymes, alteration of hormone synthesis, inhibition of cancer cell proliferation, suppression of protein expression, impede of cell cycle and impairment of cell growth. In actual circumstances active constituents present in orchidaceae family are not so explored. So, in this review we covered the article published till 2022 using different electronic databases PubMed/ MEDLINE, Scopus, Web of Science, and Google Scholar. The present review highlights the orchid plant family and its bioactive constituents with anticancer activity via various signalling pathways, as well as their possible mechanisms of action. • Cancer is a spectrum of disorders thats can leads to death. • Several orchid plants have been reported to cure various types of cancers. • Many bioactive compounds isolated from orchid plants such as Dendrobium longicornu, Dendrobium transparens, Rhyncostylish retusa, Vanda cristata, Anoectochilus formosanus etc. possess anticancer activities. • They showed anticancer activity against various types of cancer cell line. • In this study, we reviewed several anticancer properties identified in orchid plants via various signalling pathways, as well as their possible mechanisms of action.

Characterization of phenylalanine ammonia-lyase genes facilitating flavonoid biosynthesis from two species of medicinal plant Anoectochilus.

BackgroundAnoectochilus roxburghii and Anoectochilus formosanus, belong to the Anoectochilus genus, have been used for Chinese herbal drugs as well as health food. Phenylalanine ammonia-lyase (PAL), the key enzyme in primary metabolism and phenylpropanoid metabolism, produces secondary metabolites (flavonoids) in plants, which are beneficial for the biosynthesis of phenylpropanoid metabolites.MethodsThe PAL genes were cloned from A. formosanus and A. roxburghii according to our previous transcriptomic analysis. The PALs were introduced into pCAMBIA2300-35S-PAL-eGFP to generate 35S-PAL-eGFP. The constructs were further used for subcellular localization and transgenic Arabidopsis. The expression of AfPAL and ArPAL under precursor substance (L-Phe), NaCl, UV, and red-light were analyzed by real-time quantitative PCR (RT-qPCR).ResultsAfPAL and ArPAL , encoding 2,148 base pairs, were cloned from A. formosanus and A. roxburghii. The subcellular localization showed that the ArPAL and AfPAL were both localized in the nucleus with GPF. Quantitative RT-PCR analysis indicated that the ArPAL and AfPAL genes function in the phenylalanine pathway as well as response to induced conditions. Overexpression of the AfPAL and ArPAL could increase flavonoids and anthocyanin content in the transgenic Arabidopsis.DiscussionThe results suggest that AfPAL and ArPAL play a crucial role in the flavonoid biosynthesis in Anoectochilus. Also, our study provides new insights into the enrichment of secondary metabolites of traditional Chinese medicines A. formosanus and A. roxburghii, which can improve their medicinal active ingredients and be used for drug discovery in plants.

Determination of total flavonoid and polysaccharide content in Anoectochilus formosanus in response to different light qualities using hyperspectral imaging

This study demonstrated the capability of Vis/NIR hyperspectral imaging (400–1000 nm) in determining total flavonoid (TF) and total polysaccharide (TP) content in Anoectochilus formosanus that were cultivated in different light quality environments. We extracted spectra from the whole leaf (the control) and four zones (leaf tip, margin, middle and base) respectively. Partial least squares regression (PLSR) and support vector machine regression (SVMR) were used for building prediction models using the full spectral range. PLSR models outperformed SVMR models with determination of the coefficients of calibration (R2c) and validation (R2v) being over 0.9982 and the residual predictive deviations (RPD) being over 23.21. Comparing to the model for the control, the models for the four zones achieved similar performance; the model for the middle zone achieved slightly better performance than models for the other three zones (R2v = 0.9974, RMSEV = 0.14, RPD = 18.04 for TF; and R2v = 0.9982, RMSEV = 0.20, RPD = 23.56 for TP). We further selected effective wavelengths through the correlation coefficients between single wavelength, dual-wavelength combinations and measured content of TF and TP to simplify prediction models. New PLSR models were built based on the effective wavelengths with R2v, RMSEV and RPD of 0.8990, 0.85 and 3.07 for TF, and 0.8842, 1.71 and 2.79 for TP. Obtained results showed that hyperspectral imaging is promising for rapid and accurate determination of TF content and TP content in A. formosanus based on the PLSR models and the identified effective wavelengths.

Ethnomedicinal properties of orchidaceae by local communities in Lake Toba region, North Sumatra, Indonesia

Orchids are plant species with beauty and medicinal purposes, but, limited exploration has been conducted on potential utilization of Orchidaceae for medicinal purposes. This study aims to describe the local wisdom in utilization of Orchidaceae in traditional medicine and to identify the phyto-chemical content in particular orchid species which utilized for herbal medicine by local communities in the Lake Toba region. The study was conducted through exploration and in-depth interviews with 24 local communities that applied orchids in their traditional medicine in Karo, Simalungun and Toba of North Sumatra during 2020 to 2021. This study also reviewed various phyto-chemical content and safety issues of orchids utilization. The study identified about 23 orchid species from 15 genera as a main therapeutic agent. Various parts or entire plant are various parts of plant including roots, leaves, stems, flowers or entire plant contain phytochemical compounds with anti-inflammatory, anti-oxidant, antimicrobial, antipyretic properties, and applied for various medicinal. Some of well-known orchids are Anoectochilus formosanus and Calanthe triplicata. The entire plants are applied in traditional medicine to smooth the liver, to treat rheumatism, diabetes, bronchitis, wound infection, tonic or detoxification. These activities are due to various bio-active compounds, mainly alkaloids, stilbenoids, dimeric phenanthrenes, flavonoids, triterpenoids, etc. These information have been illustrated the excellent potential of Orchidaceae for novel constituents of therapeutic and herbal medicine.

Effect of lactic fermentation, microwave, and ultrasonic extraction on the bioactive compounds from Anoectochilus formosanus Hayata

Anoectochilus formosanus (family Orchidaceae) is a perennial herb that contains many bioactive valuable. In this study, the extract efficiency of bioactive compounds from Anoectochilus formosanus Hayata by microwave-assisted extraction (MAE), ultrasonicassisted extraction (UAE) treatments, and lactic fermentation in individual and combined impacts were evaluated through the total phenolic content, total polysaccharide content, and antioxidant activity. The results showed that MAE and UAE treatments and lactic fermentation enhanced the effective extraction of bioactive compounds compared to the control samples. The bioactive compound contents from A. formosanus fluid trend differences in individual treatment factors. However, there was no significant difference between these treatment factors in which the fermentation process requires more time to reach the expected extraction efficiency. The ultrasonic pretreatment combined lactic fermentation process would bring many benefits; first, no need for more the MAE treatment; second, reduce fermentation time; third, bring a probiotic source as well as valuable metabolic products from the lactic fermentation process.

Improved 1D convolutional neural network adapted to near-infrared spectroscopy for rapid discrimination of Anoectochilus roxburghii and its counterfeits

Anoectochilus roxburghii (Wall.) Lindl. (Orchidaceae) is a rare traditional Chinese medicine. For seeking high profit, some traditional Chinese medicine sellers usually adulterated A. roxburghii with Goodyera Schlechtendaliana and Ludisia discolor or directly fake A. roxburghii using Anoectochilus formosanus. These counterfeits with similar appearance greatly influence the prescription efficacy. Therefore, there is an urgent need for an effective and fast authentication method to identify A. roxburghii and its counterfeits. In this paper, the near-infrared spectroscopy (NIRS) data of A. roxburghii and its counterfeits are mearsured. Then, an improved inception architecture based 1-dimensional convolutional neural network (Improved 1D-Inception-CNN) is designed for processing the NIRS data and identifying A. roxburghii and its counterfeits. The Improved 1D-Inception-CNN has less parameters and high calculation efficiency which makes the identification model more practical. The experimental results show that compared with traditional structured CNN models, the complexity of the Improved 1D-Inception-CNN is reduced by 40 %, the parameters are reduced by 50 % and the performances are improved by 1.01 %. Therefore, the Improved 1D-Inception-CNN model based on NIRS technology can effectively and quickly identify A. roxburghii and its counterfeits.

Biologically active composite based on fumed silica and Anoectochilus formosanus Hayata extract

Biologically active extract was obtained from the leaves of Anoectochilus formosanus Hayata plants grown in vitro, and the composite was prepared by adsorption/precipitation of this extract components on the fumed silica surface under fluidized bed conditions. The composite was characterized by means of ultraviolet–visible and Fourier transform infrared spectroscopy, thermogravimetric analysis–derivative thermogravimetric analysis, scanning electron microscopy, and nitrogen adsorption method. Antioxidant properties of the extract and the composite were studied using Folin–Ciocalteu method and 2,2-diphenyl-1-picrylhydrazyl (DPPH) test; nitric oxide radical and hydroxyl radical scavenging activities of the extract and the composite were also evaluated. It was shown that adsorption/precipitation of phytochemicals does not significantly affect the texture of silica powder, while the presence of silica does not reduce the antioxidant ability of the extract components. At the same time, inclusion in the composite was found to provide bioactive substances with elongated effect and enhanced storage stability; caking resistance of silica particles was improved, as well.

An integrated analysis of metabolomic and transcriptomic profiles reveals flavonoid metabolic differences between Anoectochilus roxburghii and Anoectochilus formosanus

Anoectochilus roxburghii (Wall.) Lindl (Ar) and Anoectochilus formosanus Hayata (Af) are two species of valuable Chinese herbal medicines that belong to the genus Anoectochilus in Orchidaceae and have often been applied as the Chinese herbal medicine “Jin Xian Lian”. However, these species exhibit obvious differences in leaf color, and the difference in their medicinal value remains unclear. In this study, the metabolomes of leaf samples of Ar and Af were analyzed by high-performance liquid chromatography-mass spectrometry (LC–MS) in the positive and negative ion modes. The analysis of the leaves of both species in the positive ion mode identified 4013 metabolites enriched in 106 metabolic pathways, and the analysis in the negative ion mode found 1530 metabolites enriched in 98 metabolic pathways. A differential metabolic profiling analysis of the results obtained in positive ion mode found that 816 differentially accumulated metabolites were enriched in 80 metabolic pathways, whereas 297 differentially accumulated metabolites enriched in 55 metabolic pathways were obtained from the analysis in the negative ion mode. The number of flavonoids was higher than those of alkaloids and terpenoids. A quantitative analysis of the differentially accumulated metabolites showed that the most highly upregulated metabolites (Af/Ar) among the top 20 differentially accumulated metabolites in Af were mainly related to amino acid-related metabolic pathways, whereas the most highly downregulated metabolites (Af/Ar) in Af were mainly associated with flavonoid-related metabolic pathways. In addition, the transcriptome sequencing of Ar and Af leaf samples yielded 40.12 Gb of transcriptomic data. The analysis of differentially expressed genes (DEGs) classified 555 metabolism-related DEGs into 23 metabolism-related KEGG pathways, and the top DEGs were involved in the phenylpropanoid biosynthetic pathways, which included up to 138 DEGs associated with the flavonoid metabolism pathway. In addition, significant differences in the flavonoid-related KEGG pathways were found. A comprehensive analysis of the metabolomes and transcriptomes showed that the genes annotated as chalcone synthase (CHS), chalcone isomerase (CHI), flavonol synthase (FLS), caffeoyl-CoA O-methyltransferase (CCOAOMT) and anthocyanidin synthase (ANS) were downregulated in Af leaves compared with Ar leaves, which might account for the downregulation of flavonoid-related metabolites in Af. In summary, both the metabolomic and transcriptomic analyses showed significant differences in flavonoid metabolism between the Ar and Af leaves, and the mechanisms underlying the molecular regulation of these differences were preliminarily analyzed.

Genetic variation of Anoectochilus formosanus revealed by ISSR and AFLP analysis

Anoectochilus formosanus is an expensive medicinal folk herb in Taiwan. The plant has been used to treat hypertension, diabetes, and hepatitis by native people for a long time. In this study, 4 typical lines of A. formosanus, red-stem, green-stem, narrow-leaf, and round-leaf, were collected and screened to identify the line-specific SSR (single sequence repeat) markers. In addition, we found the narrow-leaf line exhibited the highest polysaccharide content (3.39 ± 0.74%, polysaccharide wt/plant dried wt) which is more than twofold of the lowest content of the red-stem line (1.27 + 0.41%). In the culture program construction, the extent of the somaclonal variation among 20 regenerated shoots from the same parental individual of a narrow-leaf line was revealed using the amplified fragment length polymorphism (AFLP) technique. The aim of this study is to preliminarily investigate the genetic variation of A. formosanus through polymorphic marker screening, which will benefit to the cultivation of A. formosanus with high polysaccharide content in future work. The results showed that 6 out of 81 primers screened, generating a total of 26 markers, were able to differentiate the 4 lines in the inter single sequence repeat (ISSR) analysis. On the other hand, 17 sets of AFLP primers were chosen to detect the somatic variation among the excised shoots derived from the same individual. The genetic variation among the regenerated shoots ranged from 0.00% to 5.43%. The primer pair B6 exerts the most powerful selection ability in the AFLP analysis among 20 somaclonal samples used, whereas the E4 primer pair the lowest. The AFLP primer pairs become useful tools for selecting the lowest somaclonal variation from the regenerated shoots of A. formosanus under the tissue culture program.

Evaluation of an in vitro virus culture system of anti-virus study of the Chinese herb

Influenza A virus was selected for anti-virus study with a garlic extract and a Chinese herb, Anoectochilus formosanus Hay. The virus was inoculated into the human promonocyte cell line, HL-CZ, which had been estabished in this laboratory. The infected cells have shown viral antigens detected by immunofluorescence staining with both monoclonal antibodies against virus nucleoprotein (NP) and hemagglutinin (HA) after 1-3 days of virus inoculation. This in vitro virus culture system was used as a tentative model for evaluating anti-virus activity of Chinese herbs. The relative minimum concentration of cytotoxicity (MCC) of the herb preparation to the HL-CZ cells was observed and determined during 7 days of cultivation. The MCCs tested for garlic and A. formosanus Hay extracts were 1:10 dilution (from the original package of garlic essence) (10% w/v) and 4.0 mg/ml, respectively. In several experiments, the average number of cells infected with the influenza A virus was 59.7%; those of the cells simultaneously treated with sub-MCC of garlic A. formosanus Hay and amantadine solutions were 19.4%, 33.6%, and 30.6%, respectively. These results suggest that the expression of influenza viral antigens in cells was partially inhibited by the component in these herb extracts. In addition, the replication of viral RNA was measured by polymerase chain reaction (PCR) after the HA1 region of viral hemagglutinin gene was converted into cDNA with reverse transcription. It was shown that viral HA gene could be detected 12 hours post-inoculation even in the presence of Chinese herbs. This method could be applied to detect the replication of viral RNA when the virus-infected cells were treated with the extracts of Chinese herbs.

Coculture with two Bacillus velezensis strains enhances the growth of Anoectochilus plants via promoting nutrient assimilation and regulating rhizosphere microbial community

The potentiality of PGPR and biocontrol bacteria in agriculture is steadily increased as it offers an attractive way to replace the use of chemical fertilizers, pesticides and other supplements. This study aimed to investigate the effects and potential mechanism of inoculation with two Bacillus velezensis strains on the growth and quality improvement of Anoectochilus roxburghii (Wall.) Lindl. (MRH) and Anoectochilus formosanus (Wall.) Lindl. (YYB). The control treatment was non-inoculated (CK) and the coculture treatments were inoculated with Bacillus velezensis D2WM (D2), Bacillus velezensis ZJ-11 (ZJ-11), Bacillus velezensis D2WM and Bacillus velezensis ZJ-11 (D2+ZJ-11). The fresh weight, plant length, amount of active compounds, as well as chlorophyll content were significantly increased under D2, ZJ-11, and D2+ZJ-11 treatments compared with MRH and YYB plants in CK. In particular, D2+ZJ-11 treatment resulted in the greatest growth promotion. In MRH and YYB plants, the fresh weight increased by 82.6 % and 106.6 %, the kinsenoside content increased by 9.33 % and 21.65 % per gram, and the flavonoid content increased by 44.70 % and 21.07 % per gram, respectively. D2WM and ZJ-11 were capable of secreting siderophore, phytase, indole-3-acetic acid (IAA), and zeatin to promote plant nutrient assimilation and growth. Moreover, when D2WM and ZJ-11 were both inoculated with Anoectochilus roxburghii, the rhizosphere soil enzyme activities of urease, phosphatase, and invertase were significantly higher than control, thus helped to provide more nutrients to Anoectochilus. Further, the analysis of microbial community diversity indicated the increase of abundance of beneficial microorganisms, such as nitrogen-fixing bacteria, Basidiomycota and Ascomycota both of which are known to fuel their plant partners with mineral nutrients from the soil. Additionally, the abundance of pathogens belonging to Xanthobacteraceae as well as Cladophialophora and Penicillium were reduced. Hence, coculture with beneficial microorganisms can improve the growth and quality of Anoectochilus plants via promoting nutrient assimilation and regulating rhizosphere microbial community. This technique is of great application value in promoting the efficient cultivation and production of high-quality plant materials.

Effect of calcium-alginate bead and Anoectochilus formosanus Hayata extract fluid on the viability of Lactobacillus plantarum ATCC 8014 and bioactive compounds in fermented apple juice

The aim of this study was to determinate the effect of the encapsulation by calciumalginate containing (MA sample) or non-containing Anoectochilus formosanus Hayata extracted fluid (M sample) on the survival of Lactobacillus plantarum ATCC 8014 in fermented apple juice for 60 hours. The antioxidant activity, total polyphenol, polysaccharide, pH values, and the density of L. plantarum were determined every 12 hours of fermentation. The fermented apple juice was stored at 4°C in 5 weeks. The pH value and the viable L. plantarum were evaluated during storage and in the simulation gastric medium after 4 weeks of storage. The results showed that bioactive compounds increased in the first 24 hours but decreased slowly in subsequent hours of fermentation in which the sample containing encapsulated bead had better results than free cells (F samples). The scavenging activity DPPH, total polyphenol, and polysaccharide of the MA sample were 6.58 mg Vit C/100mL; 304.65 mg GAE/100mL; and 2.98 mg Glu/100 mL, respectively. The viability of L. plantarum was maintained over 6 log CFU/mL for the encapsulated samples compared to 4 log CFU/mL for the F samples. The viability of encapsulated L. plantarum in A and MA samples was no significant difference during storage, but the survival rate of L. plantarum in MA sample was significantly higher than M samples in the SGF (Simulated gastric fluid) medium. The results indicated that adding the A. formosanus Hayata extract fluid into the calcium-alginate matrix protected L. plantarum cells during fermentation, storage and in the SGF medium.

MORFOGENESIS ANGGREK (Anoectochilus formosanus) SECARA IN VITRO

ABSTRACT Anoectochilus formosanus is one species of Orchids, known as a “Jewel Orchids” and have been used as a folk medicine in China and Taiwan.The aim of this study was to examine the response of using different types of explants and combination of growth regulators TDZ and BAP to morphogenesis of Anoectochilus formosanus Orchids by in vitro. The results showed that using different types of explants had very significant effect to the percentage of callus and shoot morphogenesis on 12 weeks after planting, the number of adventitious buds, adventitious shoot length, and callus diameter on three weeks until 12 weeks after planting. The best explant to callus morphogenesis was showing on shoot tip explant (50%) and to shoot morphogenesis was showing on auxillary buds and internode explant (100%). The best number of adventitious buds was observed in second internode explant with average number of shoots are 7.62 shoots. The best adventititous length was observed in axillary buds explant with average number of shoot length in 1.16 cm, and the best callus diameter was observed in shoot tip explant with average of diameter in 2.2 mm. The combination of plant growth regulator TDZ and BAP had a very significant effect to adventitious length on 10 weeks after planting, and a significant effect to adventitious length on 11 and 12 weeks after planting. The best adventitious length was observed on 0.25 mg L -1 TDZ + 0.75 mg L -1 BAP in 1.13 cm per explant on 12 weeks after planting. The best combination to callus and shoot morphogenesis was observed on 0.75 mg L -1 TDZ + 0.25 mg L -1 BAP. There was no interaction between used a different types of explant and combination plant growth regulator to morphogenesis Anoectochilus formosanus orchids by in vitro.

Validation of the Component Model for Prediction of Moisture Sorption Isotherms of Two Herbs and other Products

Sorption isotherm is an essential property for the processing of biological materials. In this study, a component model for the prediction of the sorption isotherm was evaluated. In order to validate this component model, the moisture sorption isotherms for Chrysanthemum morifolium flowers and the orchid Anoectochilus formosanus Hayata were determined. The sorption isotherm was measured by using the equilibrium relative humidity technique for five temperatures. Seven sorption isotherm models were selected with four quantitative criteria and residual plots to evaluate fitting ability and prediction performance for these products. The results indicated that the sorption temperature did not significantly affect the adsorption isotherm. The Caurie and Henderson equations could be used for C. morifolium flowers and A. formosanus Hayata. The isotherm data of raw bamboo, elecampe and three varieties of corn kernels from the literature were adopted to validate the component model. Comparing with the predicted values of component values and actual isotherm moisture, the component model has good predictive ability at the aw range smaller than 0.7. Considering the practical application, the aw range below 0.7 is the main range for the processing of agricultural products, and the predictive values of this component model could be helpful for food engineering and for the food industry.

Extraction of Bioactive Component from Herbal Anoectochilus formosanus Hayata by Microwave, Ultrasound and Lactic Fermentation

Anoectochilus formosanus Hayata was demonstrated to have a benefit healthy due to containing active pharmaceutical ingredients. However, A. formosanus is usually processed to produce tea bags which would destroy the bioactive compounds because of the processing procedure. The aim of this study was to evaluate the influence of extracted methods including microwave-assisted extraction (MAE), ultrasound-assisted extraction (UAE), and fermentation by Lactobacillus acidophilus ATCC-4356 to extract the active pharmaceutical ingredients from A. formosanus. The extracted liquid was analyzed total phenolics, total polysaccharide, and antioxidant activity. The results showed that three methods have a positive effect on the extraction of bioactive compounds of A. formosanus in which the fermentation showed the best result. The total phenolic content, total polysaccharide content and antioxidant capacity that extracted by the fermentation method were 11.762 mg GAE/g; 48.914 mg GE/g, and 1.582 mgVit C/g compare to MAE and UAE which were 7.818 mg and 8.128 GAE/g samples; 41.22 and 37.91mg GE/g samples; 1.032 and 1.163 mgVit C/g respectively. The A. formosanus fermentation method by L. acidophilus promotes bioactive compounds of high biological value. This study would suggest a novel use of lactic fermenting A. formosanus in the production of functional foods.

Inhibitory Effect of Anoectochilus formosanus Extract on Hyperglycemia-Related PD-L1 Expression and Cancer Proliferation.

Traditional herb medicine, golden thread (Anoectochilus formosanus Hayata) has been used to treat various diseases. Hyperglycemia induces generation of reactive oxygen species (ROS) and enhancement of oxidative stress which are risk factors for cancer progression and metastasis. In this study, we evaluated hypoglycemic effect of A. formosanus extracts (AFEs) in an inducible hyperglycemia animal model and its capacity of free-radical scavenging to establish hyperglycemia-related carcinogenesis. AFE reduced blood glucose in hyperglycemic mice while there was no change in control group. The incremental area under blood glucose response curve was decreased significantly in hyperglycemic mice treated with AFE in a dose-dependent manner. AFE and metformin at the same administrated dose of 50 mg/kg showed similar effect on intraperitoneal glucose tolerance test in hyperglycemic mice. Free-radical scavenger capacity of AFE was concentration dependent and 200 μg/ml of AFE was able to reduce more than 41% of the free radical. Treatment of cancer cells with AFE inhibited constitutive PD-L1 expression and its protein accumulation. It also induced expression of pro-apoptotic genes but inhibited proliferative and metastatic genes. In addition, it induced anti-proliferation in cancer cells. The results suggested that AFE not only reduced blood glucose concentration as metformin but also showed its potential use in cancer immune chemoprevention/therapy via hypoglycemic effect, ROS scavenging and PD-L1 suppression.

Antidiabetic activities of polysaccharides from Anoectochilus roxburghii and Anoectochilus formosanus in STZ-induced diabetic mice

Six polysaccharides were extracted from different parts (whole plants, roots and leaves) of Anoectochilus roxburghii and Anoectochilus formosanus (ARPs, ARPs-1, ARPs-2, AFPs, AFPs-1 and AFPs-2). Their primary characteristics were identified and their antidiabetic activities were evaluated in STZ-induced diabetic mice. Root polysaccharides (ARPs-1and AFPs-1) and leaf polysaccharides (ARPs-2 and AFPs-2) were distinct from each other in terms of the Mws, monosaccharide compositions and functional groups. Notably, the primary structures of ARPs and AFPs were similar to those of ARPs-1and AFPs-1, respectively. In animal experiment, after feeding with polysaccharides samples, the body weight, blood glucose, glycogen, insulin, total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), malondiadehyde (MDA) and antioxidant enzyme activities in liver and kidney of mice were tested to investigate the antidiabetic activities. All polysaccharides exhibited varying antidiabetic activities (antihyperglycemic, antioxidant and antihyperlipidemic activities), which were closely related to their primary characteristics. Furthermore, root polysaccharides with higher Mws and glucose content in both A. roxburghii and A. formosanus, exhibited better antidiabetic activities than leaf polysaccharides, and ARPs which showed the best antidiabetic activities had the potential to be used as functional food or medicine for diabetes treatment.