OBJECTIVETo study the leaf venation characteristics of Bidens pilosa and B. pilosa var. radiata and establish an identification method.

METHODLMVP (leaf morphological-venation pattern for identification Chinese herbs), and QAERM (quantitatively analyze and evaluate reliability for the method of identification Chinese herbs) were applied for the study.

RESULTUnder the transmission-light, the tertiary vein of B. pilosa was discrete, the color was darker, the size was bigger, the shape was short curve, short linear, spot-like and branch-like. However the tertiary vein of B. pilosa var. radiata was continuous linear and color lighter. With the mentioned key difference, the both plants could be successfully identified from each other, the accuracy of identification results (AC) was from 96.7% to 97.7%. The repeatability of identification results: agreement rate for observation (ARO) was 95.1% and Kappa value was 0.90.

CONCLUSIONThe established method is simple, rapid, economic and reliable.

Bidens ; anatomy & histology ; chemistry ; Pigmentation ; Plant Leaves ; anatomy & histology ; chemistry

To investigate the chemical constituents of the whole plants of Bidens bipinnata, the separation and purification of constituents were performed by chromatography on macroporous resin, silica gel, MCI and Sephadex LH-20. Their structures were elucidated by spectroscopic data as quercetin (1), quercetin-3-0-alpha-L-rhamnoside (2), keampferol-3-O-beta-D-glucopyranoside (3), keampferol-3-O-alpha-L-rhamnoside (4), 3', 5-dyhydroxy-3, 6, 4'-trimethoxyl -7-O-beta-D-glucopyranoside flavonoid (5), 7, 8, 3', 4'-tetraflavanone(6), (2S)- and (2R)-isookanin-7-O-beta-D- glucopyranoside (7a/7b), (2S)- and (2R)-3'-methoxy-isookanin-8-O-beta-D-glucopyranoside (8a/8b), 6, 7, 3', 4'-tetrahydroxyaurone(9), maritimetin (10), esculetin (11), 3-O-caffeoyl-2-methyl-d-erythrono-1, 4-lactone (12), (7S, 8R) balanophonin-4-O-beta-D-glucopyranoside (13), eugenyl-O-beta-apiofuranosyl-( 1"-6') -O-beta-glucopyranoside (14), and (+)-syringaresinol-4'-O-beta-D-glucopyranoside (15). Compounds 8, 13, 14, and 15 were isolated from this genus for the first time. Compounds 1 and 6 were potent inhibitors against HSC-T6 cells in vitro and compounds 1, 2, 6, and 7 were capable of decreasing the inflammatory cytokine production of macrophage cells in vitro.
Bidens ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

To explore the protective effect and mechanism of ethyl acetate extract from Bidens bipinnata on hepatocyte damage induced by endoplasmic reticulum stress. Tunicamycin was used to establish the damage model in L02 cells. Methyl thiazolyl tetrazolium(MTT) colorimetric assay was used to investigate the survival rate of ethyl acetate extract from B. bipinnata in L02 cells injury induced by endoplasmic reticulum stress; the protein expressions of endoplasmic reticulum stress-related molecule glucose regulated protein 78(GRP78), PKR-like ER kinase(PERK), eukaryotic initiation factor-2(eIF2α), activating transcription factor 4(ATF4), C/EBP homologous protein(CHOP), B-cell CLL/lymphoma 2(Bcl-2), Bal-2 associated X apoptosis regulator(Bax) were examined by Wes-tern blot. The expressions of the above proteins were also detected after endoplasmic reticulum stress inhibitor(4-phenyl butyric acid) and CHOP shRNA-mediated knockdowns were added. The expressions of GRP78, PERK, CHOP in L02 cells were observed by immunofluorescence method. The results showed that ethyl acetate extract from B. bipinnata could significantly increase the survival rate of L02 cell injury caused by endoplasmic reticulum stress in a dose and time-dependent manner(P<0.05 or P<0.01). The expression levels of GRP78, PERK, eIF2α, ATF4, CHOP and Bax in the drug treatment groups were significantly down-regulated(P<0.05 or P<0.01), while Bcl-2 was significantly up-regulated(P<0.01). After endoplasmic reticulum stress inhibitor and CHOP shRNA-mediated knockdowns were added, the expression levels of GRP78, PERK, eIF2α, ATF4, CHOP, Bax in the drug treatment groups were significantly down-regulated(P<0.01), whereas Bcl-2 was significantly up-regulated(P<0.01). Immunofluorescence results showed that the expressions of GRP78, PERK, CHOP were consistent with the Western blot method. In conclusion, ethyl acetate extract from B. bipinnata has a significant protective effect on the damage of L02 cells caused by endoplasmic reticulum stress. The mechanism may be related to the inhibition of endoplasmic reticulum stress and the down-regulation of apoptosis in cells through the PERK/eIF2α/ATF4/CHOP signaling pathway.
Acetates ; Apoptosis ; Bidens ; Endoplasmic Reticulum Stress ; Hepatocytes ; Transcription Factor CHOP/genetics* ; eIF-2 Kinase/genetics*

The prevalences of the fluke belonging to genus Metagonimus have been reported along the upper stream of inhabitants by several workers since 1980, however the taxonomical problems of the fluke was not yet settled. The larval flukes; cercaria and metacercaria as well as their intermediate hosts, and adult were studied in order to identify the Metagonimus in the areas. The results obtained are summarized as follows: The snails, Semisulcospira globus were collected from the three different localities along the upper stream of the River. The cercariae were found from 125(7.2%) out of 1,730 snails by natural emerging method, and were identified into 5 species including Metagonimus sp. (3.7%), Pseudexorchis major(1.4%), Cercaria nipponensis (0.9), Cercaria incerta (0.6%) and Cercaria yoshidae(0.6%). Cercariae of Metagonimus species had four to five oral spines on its anterior of the first line. The cercariae of Metagonimus were experimentally exposed to goldfish. Infection rate was 22.9% out of 105 goldfish, and the encysted metacercariae were found in fins(86.7%) and on scales (13.7%) of the fishes, but not in their muscle, head or visceral organs. Seven species of fish were caught in the Daecheong reservoir and the upper stream. Infestations with metacercaria of Metagonimus were found 100% in Opsariichtys bidens and the parasitized numbers of the metacercariae were observed from 250 to 2,400 per fish. In the upper stream, Zacco temmincki, Z. platypus and Pseudogobio esocinus were infected 100% with the metacercaria, on the other hand, the fishes caught in the reservoir showed the lower infestation rates, and a few metacercariae found in the fishes Carassius carassius and Cyprinus carpio in the reservoir and the stream. The majority of metacercariae was detected only on the scales of fishes. In order to know the infectivity and the distribution patterns in the intestine of hosts, rats and dogs were infected with the metacercariae obtained from O. bidens and Z. platypus. In addition the metacercariae obtained from Z. temmincki, P. esocinus and goldfish were given to the rats. The recovery rates of the worms in the small intestine of dogs were higher (63.3-65.8%) than those of the rats (3.5-31.6%). The flukes were found mostly in the middle and the lower part of small tntestine of the rats and the dogs, but no worm was collected in the upper part of the intestine of rats. The size of adult flukes varied by the hosts. In the adult flukes, oral sucker was smaller than ventral sucker, and the right and left testes were located diagonally, the uterine tubules circled around the upper left testis. The average egg size was 29.1 x 17.7 micro-meter. According to the above results, the flukes belonging to genus Metagonimus distributed along the Geum River was concluded to be identical with Miyata type of M. yokogawai as that Saito had proposed.
parasitology-helminth-trematoda ; Metagonimus sp. ; epidemiology ; cercaria ; metacercaria ; Zacco temmincki ; Zacco platypus ; Pseudogobio esocinus ; Opsariichtys bidens ; Carassius carassius ; Cyprinus carpio

OBJECTIVETo study the optimization process of microwave-assisted extraction for active ingredients in different parts of TMC.

METHODUsing the uniform design, the optimization processes of microwave-assisted extraction for Flavonoids and Paneonolum in Bidens and Cortex Moutan were gained.

RESULTThe optimization process of extraction for Flavonoids in Bidens can be described as following: microwave power was 850 W, radiation time was 30 min, solvent concentration was 40%, solvent volume was 13 times as the proportion of raw material, and dipping time was 60 min. The optimization process of extraction for Paneonolum in Cortex Moutan has also been gained which could be shown as following: 340 W as microwave power, 20 min as radiation time, 85% as solvent concentration, 1:5 as the proportion of raw material to solvent, and 30 min as the dipping time.

CONCLUSIONThese can prove it reasonable to get active ingredients in Bidens and Cortex Moutan with microwave-assisted extraction technology.

Acetophenones ; isolation & purification ; Bidens ; chemistry ; Flavonoids ; isolation & purification ; Microwaves ; Paeonia ; chemistry ; Plants, Medicinal ; chemistry ; Reproducibility of Results ; Technology, Pharmaceutical ; instrumentation ; methods

This study aimed to investigate the effect and possible mechanism of Bidens pilosa decoction on non-alcoholic fatty liver disease(NAFLD) induced by high fat and high glucose in mice. Bald/c mice were randomly divided into normal group, model group, metformin(200 mg·kg~(-1)) treatment group, Bidens pilosa decoction(10 g·kg~(-1)) treatment group, metformin and B. pilosa decoction(100 mg·kg~(-1)+5 g·kg~(-1)) treatment group. Except for the normal group, mice in the other four groups were fed with high-fat and high-glucose diet for 8 weeks to establish the non-alcoholic fatty liver model. After 4 weeks of treatment, blood was collected from the eyeballs, the mice were sacrificed, and relevant indicators were detected. The results showed that compared with the model group, blood lipid and blood glucose levels of each treatment group were significantly lower(P<0.05); HE staining results showed that liver pathological damage in each treatment group was significantly improved; oil red O staining results showed fat distribution in each treatment group significantly reduced(P<0.01); immunohistochemical staining showed that glucose regulated the protein expression of protein 78(GRP78) in liver tissues of each treatment group was also significantly reduced(P<0.01); Western blot results showed that endoplasmic reticulum stress signal pathway-related factors GRP78, phosphorylated-protein kinase R-like ER kinase(p-PERK), eukaryotic translation-initiation factor 2α(eIF2α), activating transcription factor 4(ATF4), C/EBP homologous protein(Chop), inositol requiring 1α(IRE1α), and cleaved-cysteinyl aspartate specific proteinase 12(cleaved-caspase-12) were significantly reduced(P<0.01). The results of the combined drug treatment group were better than those of the single drug treatment group. These results showed that B. pilosa decoction had the effect in improving non-alcoholic fatty liver, and its mechanism may be related to the down-regulation of the expression of endoplasmic reticulum stress(ERS)-related factors, and the reduction of the apoptosis of hepatocytes caused by ERS and the down-regulation of blood lipid and blood glucose levels.
Animals ; Apoptosis ; Bidens ; Endoplasmic Reticulum Stress ; Endoribonucleases ; Glucose ; Mice ; Non-alcoholic Fatty Liver Disease ; Protein-Serine-Threonine Kinases

OBJECTIVETo explore the protective effect and mechanism of total flavones of Bidens pilosa L. (TFB) on IgA1 induced injury of venous endothelial cells in Henoch-Schönlein purpura (HSP) children patients. METHODS Human umbilical venous endothelial cells (HUVECs) were taken as subject. They were intervened by normal IgA1 and HSP children patients' serum IgA1, and added with different concentrations TFB at the same time. Then they were divided into the blank control group, the normal control group, the HSP IgA1 group, and HSP IgA1 plus TFB (1.0, 0.5, 0.25 mg/mL) groups. Levels of TNF-α and IL-8 in supernate were detected by ELISA. The NO level was detected by nitrate reductase method. mRNA and protein expressions of NF-κB and ICAM-1 in HUVECs were detected by fluorescent quantitative PCR and Western blot respectively.

RESULTSCompared with the normal control group and the blank control group, levels of IL-8, TNF-α, and NO all significantly increased in the HSP group (P < 0.05). Compared with the HSP group, levels of IL-8, TNF-α, and NO significantly decreased after intervention of TFB (1.0 and 0.5 mg/mL; P < 0.05, P < 0.01). Results of fluorescent quantitative PCR and Western blot showed, as compared with the blank control group and the normal control group, mRNA and protein expressions of NF-κB and ICAM-1 in HSP children patients' serum IgA1 induced venous endothelial cells significantly increased with statistical difference (P < 0.05, P < 0.01). Compared with the HSP group, mRNA and protein expressions of NF-KB and ICAM-1 were obviously down-regulated after intervention of TFB (1.0, 0.5, 0.25 mg/mL), with statistical difference (P < 0.05, P < 0.01).

CONCLUSIONTFB could protect vascular damage by inhibiting in vivo high expression of NF-κB, reducing the production of IL-8, TNF-α, and NO in vascular endothelial cells of HSP children patients.

Bidens ; chemistry ; Child ; Flavones ; pharmacology ; Human Umbilical Vein Endothelial Cells ; drug effects ; Humans ; Immunoglobulin A ; blood ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-8 ; metabolism ; NF-kappa B ; metabolism ; Nitric Oxide ; metabolism ; Purpura, Schoenlein-Henoch ; blood ; RNA, Messenger ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism