Gu Sui Bu (Drynaria fortunei J. Sm.) antagonizes glucocorticoid-induced mineralization reduction in zebrafish larvae by modulating the activity of osteoblasts and osteoclasts.

ETHNOPHARMACOLOGICAL RELEVANCE: Gu Sui Bu (GSB), the dried rhizome of Drynaria fortunei J. Sm., is widely used in traditional Chinese medicine for treating fractures and osteoporosis. Although glucocorticoids are widely prescribed in modern medicine, the efficacy of GSB in treating glucocorticoid-induced osteoporosis (GIOP) remains unclear. AIM OF THE STUDY: GIOP is one of the most prevalent forms of osteoporosis and increases the risk of fracture, which can cause severe complications in elderly people. Safe, efficacious, and cost-effective treatment options for GIOP are thus warranted. The present study investigated the efficacy and mechanism of GSB for treating GIOP. MATERIALS AND METHODS: We established an efficient and robust in vivo GIOP model by optimizing zebrafish larvae rearing conditions and the dose and duration of dexamethasone treatment. Bone calcification was evaluated through calcein staining. To quantify the degree of vertebral mineralization in the larvae, we developed a scoring system based on the rate of vertebral calcification; this system reduced quantification errors among individual zebrafish caused by inconsistencies in staining or imaging parameters. Quantitative real-time polymerase chain reaction was used to access the expression levels of genes essential to the differentiation and function of bone cells. High-performance liquid chromatography was employed to identify naringin in the GSB extract. RESULTS: GSB significantly reversed the dexamethasone-induced calcification delay in zebrafish larvae. GSB enhanced osteoblast activity by increasing the expression of collagen I, osteopontin, and osteonectin and repressed bone resorption by decreasing the expression of matrix metalloproteinases (mmps), including mmp9 and mmp13a. We also identified naringin as one of the constituents of GSB responsible for the herbal extract's anti-GIOP activity. CONCLUSIONS: Using the in vivo zebrafish GIOP model that we established, the efficacy of traditional Chinese medicines in treating GIOP could be systematically investigated. GSB has an osteogenic effect and may thus be an efficacious and cost-effective treatment option for GIOP. Notably, bone resorption activity was found to be retained after GSB treatment, which would be beneficial for maintaining normal bone remodeling.

Drynaria fortunei improves lipid profiles of elderly patients with postmenopausal osteoporosis via regulation of Notch1-NLRP3 inflammasome-mediated inflammation.

BACKGROUND: Dyslipidemia is a common comorbidity in elderly patients with postmenopausal osteoporosis (PMOP). Drynaria fortunei (Rhizoma drynariae) is well-known in traditional Chinese medicine for its ability to improve bone mineral density (BMD). However, whether and how Drynaria fortunei improves plasma lipid profiles in elderly PMOP patients remains unclear. METHODS: Eighty elderly female patients with concurrent PMOP and hyperlipemia were randomly assigned to Drynaria fortunei 2(n = 40) or control (n = 40) groups. The clinical efficacies of Drynaria fortunei were evaluated. At 0, 3-, 6-, 9-, and 12-month of follow-up, plasma levels of IL-1ß, IL-18, TNF-α, IL-6, IL-8, and IL-10 were measured using ELISA, whereas PBMC levels of NLRP3, ASC, caspase-1, NF-κB, SIRT1, and Notch1 were measured using RT-qPCR. PBMC isolated from PMOP patients were cultured and treated with Drynaria fortunei to determine its influence on NLRP3 inflammasome and associated cytokines. RESULTS: Drynaria fortunei effectively improved patients' BMD and lipid profiles. IL-1ß, IL-18, TNF-α, IL-6, IL-8 levels, as well as inflammasome-molecules of NLRP3, ASC, caspase-1, and NF-κB increased over time in the control group, but were significantly attenuated with Drynaria fortunei administration. In vitro, Drynaria fortunei suppressed NLRP3 inflammasome and associated cytokines by increasing SIRT1 or decreasing Notch1. Drynaria fortunei had inhibitory effects on NLRP3 inflammasome and Notch1 even when SIRT1 expression was suppressed. CONCLUSIONS: Drynaria fortunei has been demonstrated to significantly improve lipid profiles for elderly PMOP patients. Drynaria fortunei may down-regulate Notch1 independently of SIRT1 to suppress NLRP3 inflammasome-mediated inflammation, thus improving plasma lipid profile.

A polysaccharide from the dried rhizome of Drynaria fortunei (Kunze) J. Sm. prevents ovariectomized (OVX)-induced osteoporosis in rats.

In the present study, a homogenous polysaccharide (DFPW) was isolated and purified from the dried rhizome of Drynaria fortunei, and its protective effect against osteoporosis was investigated in ovariectomized (OVX) rats. Histological analysis indicated that oral administration of DFPW (100 and 400 mg/kg) for 12 weeks significantly improved trabecular bone mass, as demonstrated by the increase in trabecular area, trabecular thickness and its number in OVX rats. Furthermore, the decline of bone mineral density and bone mineral content including Ca, P and Mg induced by OVX was reversed by the DFPW administration. This function was achieved by the decreased levels of the bone turnover markers, such as serum ALP, urinary deoxypyridinoline (DPD), Ca and P excretions. Besides, DFPW improved biomechanical parameters (maximum load, energy, Young's, modulus and maximum stress) to strengthen the hardness and strength femoral diaphysis in OVX rats. These results strongly suggested that DFPW might be a hopeful alternative therapeutics to treat postmenopausal osteoporosis.

Combined Treatment with Two Water Extracts of Eleutherococcus senticosus Leaf and Rhizome of Drynaria fortunei Enhances Cognitive Function: A Placebo-Controlled, Randomized, Double-Blind Study in Healthy Adults.

We previously found that the water extract of Eleutherococcus senticosus leaves (ES extract) enhanced cognitive function in normal mice. Our study also revealed that the water extract of rhizomes of Drynaria fortunei (DR extract) enhanced memory function in Alzheimer's disease model mice. In addition, our previous experiments suggested that a combined treatment of ES and DR extracts synergistically improved memory and anti-stress response in mice. Although those two botanical extracts are expected to be beneficial for neuropsychological function, no clinical data has ever been reported. Therefore, we performed a placebo-controlled, randomized, double-blind study to evaluate cognitive enhancement and anti-stress effects by the intake of a combined extract in healthy volunteers. The intake period was 12 weeks. The Japanese version of the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS) test was used for neurocognitive assessment. The combined treatment of ES and DR extracts significantly increased the figure recall subscore of RBANS (p = 0.045) in an intergroup comparison. Potentiation of language domain ((p = 0.040), semantic fluency (p = 0.021) and figure recall (p = 0.052) was shown by the extracts (in intragroup comparison). In anti-stress response, the anxiety/uncertainly score was improved by the extract in an intragroup comparison (p = 0.022). No adverse effects were observed. The combined treatment of ES and DR extracts appear to safely enhance a part of cognitive function in healthy adults.

Evaluation of selected traditional Chinese medical extracts for bone mineral density maintenance: A mechanistic study.

OBJECTIVE: To investigate the development of a minimal traditional Chinese medicine (TCM) formula using selected TCM ingredients and evaluating their biological activity with bone-specific in vitro tests. Finally, determining if the minimal formula can maintain bone mineral density (BMD) in a low bone mass (LBM)/osteoporosis (OP) model system. METHODS AND RESULTS: Sixteen different TCM plant extracts were tested for estrogenic, osteogenic and osteoclastic activities. Despite robust activation of the full-length estrogen receptors α and ß by Psoralea corylifolia and Epimedium brevicornu, these extracts do not activate the isolated estrogen ligand binding domains (LBD) of either ERα or ERß; estrogen (17-ß estradiol) fully activates the LBD of ERα and ERß. E. brevicornu and Drynaria fortunei extracts activated cyclic AMP response elements (CRE) individually and when combined these ingredients stimulated the production of osteoblastic markers Runx2 and Bmp4 in MC3T3-E1 cells. E. brevicornu, Salvia miltiorrhiza, and Astragalus onobrychis extracts inhibited the Il-1ß mediated activation of NF-κß and an E. brevicornu/D. fortunei combination inhibited the development of osteoclasts from precursor cells. Further, a minimal formula containing the E. brevicornu/D. fortunei combination with or without a third ingredient (S. miltiorrhiza, Angelica sinensis, or Lycium barbarum) maintained bone mineral density (BMD) similar to an estradiol-treated control group in the ovariectomized rat; a model LBM/OP system. CONCLUSION: A minimal formula consisting of TCM plant extracts that activate CRE and inhibit of NF-κß activation, but do not behave like estrogen, maintain BMD in a LBM/OP model system.

Drynaria fortunei Promoted Angiogenesis Associated With Modified MMP-2/TIMP-2 Balance and Activation of VEGF Ligand/Receptors Expression.

Background and Purpose: Drynaria fortunei J. Sm (D. fortunei), known as Gu-Sui-Bu, is used in traditional Chinese medicine to treat common injuries, including bone fractures and bruising. The specific functional mechanisms of the angiogenic and endothelial cell migration properties of D. fortunei are currently unclear. Thus, the purpose of this study is to validate the potential angiogenic and cellular migration properties and related mechanisms by D. fortunei both in vivo and in vitro. Experimental Approach: The present study investigates, both in vivo and in vitro, the wound healing effects of D. fortunei as associated with angiogenesis, specifically by the modulation of matrix metalloproteinases (MMPs) and upregulation of vascular endothelial growth factor (VEGF) ligand/receptors. In order to determine the potential angiogenic effects of D. fortunei, in vivo neovascularization of chick chorioallantoic membranes (CAMs) assay, and directed in vivo angiogenesis assay (DIVVA) were performed, while in vitro scratch wound healing, migration, and matrix-induced tube formation assays were performed by using human umbilical vascular endothelial cells (HUVECs). Furthermore, we used qPCR to analyze the gene expressions and Western blot to observe protein expressions of MMP-2, MMP-14, TIMP-2, RECK, and VEGF/VEGFRs. Results: This study identified five major compounds from the water extract of D. fortunei: protocatechuic acid, caffeic acid 4-O-ß-D-glucopyranoside, 5,7-dihydroxychromone-7-O-rutinoside, neoeriocitrin, and naringin. D. fortunei was confirmed to activate in vivo angiogenesis by CAM and DIVVA assays. D. fortunei further exhibited in vitro angiogenic effects associated with cell migration, as demonstrated by the tube formation assay, transwell migration assay, and scratch wound healing assay. The extracellular MMP-2 activity was found to be dose-dependently augmented both in vitro and in vivo by D. fortunei. The mRNA and protein expressions of MMP-2, and MMP-14 were increased; while the tissue inhibitor metalloproteinase-2 (TIMP-2), and reversion-inducing cysteine-rich protein with kazal motifs (RECK) were both decreased. Furthermore, D. fortunei activated the gene and protein expressions of VEGF-A, -B, and VEGFR-2, -3. Conclusion: D. fortunei increased MMP-2 activity, thereby stimulating angiogenesis and cell migration, both in vivo and in vitro, as a result of MMP-2 and TIMP-2 balance modulation and the activation of VEGF/VEGFRs expression.

Immunohistopathologic evaluation of Drynaria fortunei rhizome extract in the management of Class II furcation defects in a canine model.

BACKGROUND: Management of furcation defects is still a challenging subject in periodontal therapy. Drynaria fortunei (Df) is a common type of traditional Chinese herb in the area of orthopedics and traumatology. In vitro and tissue engineering studies have shown that Df induces osteoblastic proliferation and promotes the differentiation of human periodontal ligament cells. This study investigated the management of Class II furcation defects in dogs using guided tissue regeneration (GTR) and Df granules mixed with ß-tricalcium phosphate (ß- TCP) alloplast. METHODS: Sixteen Class II critical-sized furcation defects were surgically created in four mongrel dogs: Eight defects were treated with GTR and Df granules mixed with (ß-TCP) alloplast served as the experimental group, while the other eight were managed with GTR and alloplast, served as control. Dogs were sacrificed at 4 and 8 weeks and the premolars were processed for the evaluation of treatment outcome including; osteoblastic count (OC), cementum layer thickness (CLT), percentage of collagen in bone matrix (CBM), and alkaline phosphatase (ALP) immunoreaction. RESULTS: Experimental group treated with Df showed a significant increase (P < 0.001) in the values of OC, CLT, CBM, and ALP immunoreactivity when compared with control at 4 and 8 weeks after treatment. CONCLUSION: Drynaria fortunei demonstrated increased regeneration and bone formation when used in the treatment of furcation defects in a canine model.

[Osteopractic total flavone promoting rat extra-articular tendon-bone healing through mTOR pathway].

OBJECTIVE: To explore function and related molecular mechanism of osteopractic total flavone (OTF) on tendon healing in rats. METHODS: Ten male rats aged for 8 weeks were collected and weighted from 180 to 220 g. Tendon stem cells were cultivated, the third tendon stem cells were used for experiment. OTP treated with 0, 0.1, 1, 10 ng/ml were added into tendon stem cells, and expression change of ALP, Runx2, OCN, VEGF, P-S6, P-4E/BP1 were detected after 14 days. Forty male rats aged for 8 weeks (weighted 180 to 220 g) were established extra-articular tendon-bone transplanting healing model, and divided into experimental group and control group. Experimental group were treated with OTF(100 mg·kg⁻¹·d⁻¹), while control group was treated by normal saline with the same volume. Tendon-bone healing degree were detected by biomechanical testing at 3 and 6 weeks after surgery, histological detection were applied to detect tendon-bone healing and number of new vessles. RESULTS: After treated by OTP, ALP staining and active index detection showed there were statistical differences among 0, 0.1, 1, 10 ng/ml group. After 14 days' cultivation, western blotting results showed mTOR downstream marker protein P-S6 protein expression were gradually increased with increase of density of OTP, expression of P-4E/BP1 was reduced, while expression of Runx2, OCN, VEGF were increased. Biological detection results showed that there was no significant difference in mechanical strength between experimental group(0.78±0.05) N/mm and control group (0.51±0.02) N/mm at 3 weeks after surgery, while mechanical strength in experimental group (1.36±0.09) N/mm was higher than control group (1.01±0.08) N/mm at 6 weeks after surgery. Histological results showed maturity of tendon-bone surface cell were higher at 3 and 6 weeks in experimental group, sharpey fiber growth more density, calcification extent of mesenchyme was high, and new bone, vessels were increased. CONCLUSIONS: OTF could promote osteogenic differentiation of tendon stem cells through mTOR signaling in vitro, and stimulate tendon-bone healing in bone tunnel and enhance connection quality between tendon and bone.

Chemical Constituents of Drynaria fortunei and Their Protective Effects on PC12 Cell / 中国药学杂志

OBJECTIVE: To study the chemical constituents from the rhizome of Drynaria fortunei and the protective effects of them on PC12 cells induced by Aβ25-35. METHODS: The compounds were isolated and purified by silica gel, Sephadex LH-20, ODS column chromatography, and their structures were identified on basis of spectroscopic METHODS:, such as MS and NMR. PC12 cells were treated with Aβ25-35 to establish the Alzheimer' s disease models. The compounds of different concentrations were added into culture medium to detect the protection. MTT assay was used to detect cell vitality and to observe the protective effects of compounds on PC12 cells induced by Aβ25-35. RESULTS: Nine compounds were isolated and identified as naringin(1), neoeriocitrin(2), 5,7-dihydroxychromone-7-neohesperidoside(3), (E)-4-O-β-D-glucopyranosyl caffeic acid(4), kaempferol(5), luteolin(6), protocatechoic acid(7), psoralen(8), and β-sitosterol(9). The cell experiments were performed on the compounds 1-8 and the RESULTS: showed they can promote the proliferation of PC12 cells. The cell vitality increase with concentration rising, and the difference is statistically significant (P<0.05). CONCLUSION: Compounds 1-8 play an important role in protecting Aβ25-35-induced injury in PC12 cells and they are the main active components of Drynaria fortunei in the protection of central nervous function.

Osteopractic total flavone promoting rat extra-articular tendon-bone healing through mTOR pathway / 中国骨伤

<p><b>OBJECTIVE</b>To explore function and related molecular mechanism of osteopractic total flavone (OTF) on tendon healing in rats.</p><p><b>METHODS</b>Ten male rats aged for 8 weeks were collected and weighted from 180 to 220 g. Tendon stem cells were cultivated, the third tendon stem cells were used for experiment. OTP treated with 0, 0.1, 1, 10 ng/ml were added into tendon stem cells, and expression change of ALP, Runx2, OCN, VEGF, P-S6, P-4E/BP1 were detected after 14 days. Forty male rats aged for 8 weeks (weighted 180 to 220 g) were established extra-articular tendon-bone transplanting healing model, and divided into experimental group and control group. Experimental group were treated with OTF(100 mg·kg⁻¹·d⁻¹), while control group was treated by normal saline with the same volume. Tendon-bone healing degree were detected by biomechanical testing at 3 and 6 weeks after surgery, histological detection were applied to detect tendon-bone healing and number of new vessles.</p><p><b>RESULTS</b>After treated by OTP, ALP staining and active index detection showed there were statistical differences among 0, 0.1, 1, 10 ng/ml group. After 14 days' cultivation, western blotting results showed mTOR downstream marker protein P-S6 protein expression were gradually increased with increase of density of OTP, expression of P-4E/BP1 was reduced, while expression of Runx2, OCN, VEGF were increased. Biological detection results showed that there was no significant difference in mechanical strength between experimental group(0.78±0.05) N/mm and control group (0.51±0.02) N/mm at 3 weeks after surgery, while mechanical strength in experimental group (1.36±0.09) N/mm was higher than control group (1.01±0.08) N/mm at 6 weeks after surgery. Histological results showed maturity of tendon-bone surface cell were higher at 3 and 6 weeks in experimental group, sharpey fiber growth more density, calcification extent of mesenchyme was high, and new bone, vessels were increased.</p><p><b>CONCLUSIONS</b>OTF could promote osteogenic differentiation of tendon stem cells through mTOR signaling in vitro, and stimulate tendon-bone healing in bone tunnel and enhance connection quality between tendon and bone.</p>

The effect and mechanism of naringin on callus angiogenesis of ovariectomized rats' fractures / 中华骨科杂志

Objective To explore the angiogenesis&neovascularization effects of naringin treatment in ovariectomized rats’fracture healing. Methods Upper 1/3 transverse tibial fracture model 4 weeks later after ovariectomized were estimated and randomly divided into the naringin group and control group. Microfil perfusion technique was used to analysis the angiogene?sis situation at two weeks after bone fracture. HE staining was used to evaluate the level of angiogenesis&neovascularization of tis?sue from histological point of view. The relative expression of VEGF in the callus was identified by real?time polymerase chain re?action. Immunohistochemical technique was used to observe the vessel endothelial growth factor?2 in the callus of the two groups. Maximum fracture load was tested by three?point bend test. Results The vascular volume and vascular density were more in nar?ingin group than control group. The HE staining of the 2 week group slices shows that the VA, VN2 of the unit of high magnifica?tion vision of the naringin group was significantly larger compared to the control group. Real?time PCR revealed that the compara?tive expression of VEGF is more in naringin group than in control group; the positive number of VEGFR?2 is more in naringin group than in control group. Naringin can promote the maximum load of the callus. Conclusion Naringin can promote ovariecto?mized rats’angiogenesis&neovascularization in the early process of fracture healing. It may be act on the signaling pathway of VEGF/VEGFR?2.

Triglyceride accumulation inhibitory effects of new chromone glycosides from Drynaria fortunei.

Two new chromone glycosides, drynachromosides C (1) and D (2), along with five known chromones (3-7), were isolated from the rhizomes of Drynaria fortunei. The structures of the two new compounds were elucidated on the basis of physico-chemical property and spectroscopic data. Triglyceride (TG) accumulation inhibitory effects of the obtained chromones on 3T3-L1 cells were investigated. The results showed that 1, 2 and 5 exhibited inhibitory activity on TG accumulation. Effects of compounds 1 and 2 on mRNA expression of PPARγ, C/EBPα and aP2 in 3T3-L1 cells were also investigated.

Drynaria fortunei J. Sm. improves the bone mass of ovariectomized rats through osteocalcin-involved endochondral ossification.

AIM OF THIS STUDY: Our previous study showed that Drynaria fortunei J. Sm. (Kunze), a traditional Chinese medical herb, can promote osteoblast differentiation and maturation. This study was further aimed to confirm the traditional effects of Kunze on the bone mass of ovariectomized rats. MATERIALS AND METHODS: Female Wistar rats were given an ovariectomy and then administered the water extract of Kunze (WEK). Systemic and tissue toxicities of WEK were assessed. A biomechanical test, bone mineral contents, and bone histomorphometry were analyzed to determine the effects of the WEK on the bone mass. Levels of osteocalcin (OCN) in bone tissues were determined by immunohistochemistry and immunoblotting. The effects of naringin, one of the bioactive compounds of the WEK, on the bone mass were evaluated. RESULTS: A bilateral ovariectomy in rats caused a time-dependent decrease in levels of serum 17ß-estradiol. Exposure of ovariectomized rats to the WEK at 0.5 and 1g/kg body weight/day for 1, 2, 3, and 6 months did not induce systemic or tissue toxicities. Biomechanical testing and a bone mineral content analysis showed that the ovariectomy decreased the bone torsion force and bone ash in time-dependent manners. In comparison, after exposure to the WEK, the ovariectomy-induced reductions in the bone torsion force and bone ash were significantly alleviated. In parallel, results of a bone histomorphometric assay further revealed that the ovariectomy caused significant diminution in the production of prehypertrophic chondrocytes and trabecular bone but enhanced hypertrophic chondrocyte numbers in the growth plate. However, exposure to the WEK lowered ovariectomy-induced changes in these cellular events. As to the mechanism, the WEK increased OCN biosynthesis in bone tissues of ovariectomized rats. Administration of naringin to ovariectomized rats caused significant amelioration of the bone strength, bone mineral contents, and trabecular bone amounts. CONCLUSIONS: This study shows that the WEK can translationally promote the bone mass in ovariectomized rats through stimulating OCN-involved endochondral ossification.

On the inhibitory effect of Drynaria fortunei extract on human myeloma SP20 cells.

The objective of the study was to investigate the inhibitory effect of Drynaria fortunei extract on human myeloma SP2 cells. Three different total extracts of Drynaria fortunei were obtained by reflux extraction method using different organic solvents including ethanol, methanol and petroleum ether. Their anticancer activities on SP20 cells were tested, and the maximum inhibition rate was obtained. The inhibitory effects on tumour cells at 12 h, 24 h, 36 h and 48 h were tested, and the inhibition curves at different time periods were plotted. The results showed that the methanol and ethanol extracts have similar inhibition rates at 24 h, which are around 55%. On the other hand, the maximum inhibition rate of petroleum ether extract is only 36% within 24 h. Moreover, within the time periods of 36 h and 48 h, its inhibition rates are all below 10%.

Lignans and flavonoids from rhizome of Drynaria fortunei / 中草药

Objective: To study the chemical constituents from the rhizome of Drynaria fortunei. Methods: Silica gel, ODS, Sephadex LH-20 column chromatography, and semi-preparative HPLC were used to isolate pure compounds. The compounds were identified on the basis of their physicochemical properties and spectroscopic data. Results: Fourteen compounds were isolated from the rhizome of D. fortunei, including three lignans (1-3) and eight flavonoids (4-11). By spectroscopic techniques, such as 1H-NMR, 13C-NMR, and ESI-MS, these compounds were identified as (7′R,8′S)-dihydrodehydrodiconiferyl alcohol 4′-O-β-D-glucopyranoside (1), lariciresinol 4′-O-β-D-glucopyranoside (2), (-)-secoisolariciresinol 4-O-β-D-glucopyranoside (3), eriodictyol (4), eriodictyol 7-O-β-D-glucopyranoside (5), neoeriocitrin (6), naringin (7), luteolin (8), luteolin 7-O-β-D-glucopyranoside (9), luteolin 8-C-β-D- glucopyranoside (10), 2′,4′-dihydroxydihydrochalcone (11), maltol 3-O-β-D-glucopyranoside (12), β-sitosterol (13), and daucosterol (14). Conclusion: Compounds 1-3, and 11 are isolated from the plants in Polypodiaceae family for the first time, and compounds 5 and 8-10 are isolated from the plants of Drynaria (Bory) J. Sm. for the first time. Compound 1 is present as rotamers at room temperature.

Antimicrobial Activity of Extract and Fractions from Drynaria fortunei Against Oral Bacteria

One of the traditional Korean medicine, Drynaria fortunei (D. fortunei) is one of candidates known to be effective for the treatment of inflammation, hyperlipemia, arteriosclerosis, rheumatism, and gynecological diseases such as osteoporosis and bone resorption. The present study investigated the antimicrobial activity of methanol (MeOH) extract and n-butanol (n-BuOH), chloroform (CHCl3), and ethyl acetate (EtOAc) fractions of D. fortunei against oral bacteria. The n-BuOH and CHCl3 fractions (MICs, 0.0078 to 0.3125 mg/ml; MBCs, 0.019 to 0.625 mg/ml) were demonstrated as strong antibacterial activity than the MeOH extract and EtOAc fraction. The combination effects of n-BuOH fraction with ampicillin or gentamicin were synergistic against some oral bacteria. We suggest that D. fortunei could be employed as a natural antibacterial agent in oral care products.

Study on acute toxicity and analgesic, sedative effects of Drynaria fortunei - Polypodiaceae on experimentation

The acute toxicity and analgesic, sedative effects of Drynaria fortunei - Polypodiaceae were studied on experimental mice and determined LD50 by Litchfield-Wilcoxon method. Results: this drug in dosage of 4g/kg has analgesic and sedative effect. It hasn't acute toxicity in dosage 64-fold treating dosage

Study on the anti inflammatory effect of cot toai bo in experimental animals

The anti-flammatory effect of Drynaria fortunei was studied on rat’s hind paw edema induced by carrageenin. Results showed that its aqueous extract in the dose of 4g/kg body weight exerts anti-inflammatory effect. Acute effect of Drynaria fortunei is exhibited through the decreased quantity of exudation and the number of leucocyte in the exudation.

Preparation of neoeriocitrin and naringin reference substances from Drynaria fortunei / 中草药

Objective To establish a separation method for neoeriocitrin and naringin reference sub- stances from Drynaria fortunei.Methods The extract of D.fortunei was isolated and purified by macro porous resin,silica gel column chromatography,neutro-aluminum oxide adsorbing,Sephadex LH-20 co- lumn chromatography,and recrystallizations.The structures of neoeriocitrin and naringin were identified by MS,IR,UV,~1H-NMR,and ~(13)C-NMR.Results The contents of neoeriocitrin and naringin were 99.5% and 99.3%,respectively.Conclusion The developed method is simple with lower cost,by which neoeriocitrin and naringin can be used as reference substances for the qualitative and quantitative analysis of Chinese herbal medicine.