Effect of Semen vaccariae and Taraxacu mogono on Cell Adhesion of Bovine Mammary Epithelial Cells

The aim of this study is to reveal the regulation mechanism of the effect of Semen vaccariae and Taraxacu mogono on the cell-cell adhersion molecule, E-cadherin and β-catenin on the proliferation role and secretion function of bovine mammary epithelial cells cultured in vitro. Firstly, the epithelial character of bovine mammary epithelial cells was authenticated using immunofluorescence, then the cell grow curve was observed and investigated after S. vaccariae and T. mogono treatment. On the effect of S. vaccariae and T. mogono, cell adhesion molecules E-cadherin, β-catenin and CycinD1 mRNA and protein were detected by qRT-PCR and Western blotting, respectively. The results showed that the cellular keratin 18 expressed positively and proliferfated vigorously after S. vaccariae and T. mogono treament. The mRNA and protein levels of E-cadherin and CycinD1 were remarkably higher （P〈0.05） in 36 h after S. vaccariae and T. mogono treatment. The cell proliferation at 36 h was increased significantly （P〈0.05）. In conclusion, S. vaccariae and T. mogono have a positive impact on the cell proliferation and an effect on the adhesion molecules E-cadherin, β-catenin and CycinD1 in the Wnt signaling pathway.

Influence on Cellular Signal Transduction Pathway in Dairy Cow Mammary Gland Epithelial Cells by Galactopoietic Compound Isolated from Vaccariae segetalis

The galactopoietic mechanism of Vaccaria segetalis is still unknown. Understanding dibutyl phthalate （DBP） separated from Vaccaria segetalis on the expression of lactation signal transduction genes of mammary gland epithelial cells, including prlr, erα, akt1, socs2, pparγ and elf5, will be helpful to reveal the molecular mechanism. Western blot and qRT- PCR were used to study the change of prlr, erα, akt, socs2, pparγ, and elf5 expression at mRNA and protein level. Co- localization expression of prolactin receptor （PRLR） and estrogen receptor α （ERα） was observed by immunofluorescence; the expression changes of miRNAs （21, 125b, 143, and 195） and the secretion of β-casein and lactose were detected by qRT-PCR and RP-HPLC. The results showed that Vaccaria segetalis active compound had similar fuctions as estrogen and/or prolactin （PRL） in dairy cow mammary gland epithelial cells （DCMECs）, increased the expressions of prlr, erα, akt1, and elf5 genes, while repressed pparγ expressions. DBP promoted socs2 mRNA expression, but its protein expressions were repressed. Furthermore, both DBP and PRL could repress the expressions of miRNA-125b, miRNA-143 and miRNA- 195 in DCMECs. DBP could repress the expression of miRNA-21, while the influence of PRL on miRNA-21 was not certain. DBP could promote the lactation ability of DCMECs by regulating the ER and PRLR cellular signal transduction pathway.

Metabolic Regulation of Mammary Gland Epithelial Cells of Dairy Cow by Galactopoietic Compound Isolated from Vaccariae segetalis

In previous experiment, we isolated a compound dibutyl phthalate （DBP） from Vaccaria segetalis which had galactopoietic function on mammary gland epithelial cells of dairy cow （DCMECs）. In this experiment, we ascertained the metabolic regulation function of DBP on DCMECs. Many genes related to lactation including Stat5, AMPK, b-casein, Glut1, SREBP-1, PEPCK, and ACC were detected by real-time PCR. Furthermore, Stat5 and AMPK were detected by Western blot and immunofluorescence co-localization, respectively. The results showed that DBP stimulates the expression of Stat5 and p-Stat5, thus activates Stat5 cell signal transduction pathway and stimulates b-casein synthesis. DBP also raises the activities of Glut1 and AMPK to stimulate glucose uptake and glycometabolism and activates the expression of AMPK downstream target genes PEPCK and ACC and expression of SREBP-1 to stimulate milk fat synthesis. In addition, the activities of HK, G-6-PDH, ICDH, ATPase, and energy charges were stimulated by DBP to increase the energy metabolism level of DCMECs. The results showed DBP stimulates energy metabolism related to galactopoietic function in DCMECs.

Segetoside L, A New Triterpenoid Saponin from Vaccaria segetalis

A new triterpenoid saponin, named segetoside L, was isolated from the seeds of Vaccaria segetalis. On the basis of spectral data and chemical reaction, its structure was established as 28-O-b-D-glucopyranosyl-(16)-b-D-glucopyranosyl oleanolic acid 3-O-b-D- glucopyranosyl-(13)-[b-D-galactopyranosyl-(12)]-b-D-galactopyranoside.

A New Triterpenoid Saponin from the Seeds of Deccaria segetalis

A new triterpenoid saponin, named segetoside k, has been isolated from the seeds of Vaccaria segetalis. On the basis of chemical reaction and spectral analysis, the structure of segetoside K was established as:olean-12-ene-23a, 286-dioic acid 36, 16a-dihydroxy-28-O- [βD-glucopyranosyl-(1 →3)]-β-D-glucopyra-nosyl-(l →2)-β-D-glucopyranosyl-(l→6)-β-D- glucopyranoside(1).

OBSERVATION ON 30 CASES WITH TEMPOROMANDIBULAR ARTICULAR DYSFUNCTION SYNDROME TREATED BY AURICULAR-POINT-PRESSING THERAPY

The writer has treated 30 cases of temporomandibular articular dysfunction syn-drome with only auricular pressing therapy. The method: A seed of vaccaria segetalis is used to tapethe Antitratic Apex point and the Shenmen point of ear, then the practitioner presses the seed withhand to produce sensations, such as distension, numbness, heat, and a little sweating etc.. Exchangtwo sides of the ears alternatively. The treatment is performed once every two or three days and threetimes are constituted one course. In one or three courses we can obtain the therapeutic effectiveness as50% cured, 43% improved and 6.7% failed.

Proteomic Identification of Differentially Expressed Proteins in Vaccaria segetalis-treated Dairy Cow Mammary Epithelial Cells

To investigate the potential effects of Vaccaria segetalis, we employed the proteomic approach on the dairy cow mammary gland epithelial cells （DCMECs）. A total of 35 differentially expressed nuclear proteins were visualized by 2-DE and silver nitrate staining. Of these, five proteins also displayed significant expression changes upon treatment of the water decoction from Vaccaria segetalis, and such alterations were further confirmed by RT-PCR. Together, at both the mRNA and protein levels, the water decoction from Vaccaria segetalis increased the expression of proteins ethylmalonic encephalopathy 1 （ETHE1）, vesicle amine transport protein 1 homolog （VAT1）, parkinson disease protein 7 homolog （Protein DJ-1）, proteasome subunit alpha type-2 （PSMA2） and SUMO-activating enzyme subunit 1 （SAE1）. This study would enable a better understanding of the molecular mechanisms underlying this water decoction effects at the protein level.

Galactopoietic Activity of Dibutyl Phthalate Isolated from Vaccaria segetalis

A galactopoietic compound, identified as dibutyl phthalate（DBP）, was isolated from Vaccaria segetalis. The activity of DBP on lactation ability of dairy cow mammary gland epithelial cells（DCMECs） cultured in vitro and dairy cow was evaluated. Results showed that DBP could promote cell viability, proliferation ability, lactose and β-casein secretion of DCMECs, which could also raise the milk yields of dairy cows significantly.

Vaccariside A, Novel Saponin from Vaccaria segetalis (Neck.) Garcke

A new triterpenoid saponin, vaccariside A 1 was isolated from the seeds of l'accaria segetalis using various chromatographic methods. The structure of compound 1 was elucidated by comprehensive spectroscopic analysis as 3-O-beta-D-galactopyranosyl-(1-2)-[beta-D-quinovpyranosyl-(1-4)-beta-D-xylopyranosyl-(1-3)]-beta-D-glucuronopyranosyl quillaic acid 28-O-beta-D-glucopyranosyl-(1-3)-[beta-D-xylopyranosyl-(1-4)]-alpha-L-rhamnopyranosyl-(1-2)-3-O-acetyl-beta-D-fucopyranoside.

A comparative study on the anti-bacterial effect of Echinops persicus, Cardamine uliginosa and Vaccaria oxyodontha Boiss on Staphylococcus aureus and Escherichia coli bacteria in vitro

Background:Medicinal herbs have always been closely associated with human beings,and no one deny their medicinal effects and uses.There are over 7,500 plant species in the rich flora of Iran.This study,intends to investigate the anti-bacterial properties of three different plant species from Iranian flora,including Echinops persicus,Cardamine uliginosa,and Vaccaria oxyodontha Boiss.Materials and methods:This cross-sectional study was performed after confirmation and identification of collected plants.The plants were dried in the shade for extraction.Then the methanol extract of these plants was extracted by the classical extraction method.The plants’antimicrobial effects of the plants were evaluated by agar disk diffusion,minimum inhibitory concentration,and minimum bactericidal concentration.The mentioned plants’anti-bacterial activity was compared with the antibiotic discs of gentamicin(10 mg)and vancomycin(30 mg).SPSS software v20 was used for data analysis of Kruskal-Wallis and Dunn’s tests(P<0.05).Results:The lowest minimum inhibitory concentration obtained from the methanol extract of Cardamine uliginosa on standard strains of Staphylococcus aureus and Escherichia coli at 15.62 mg/mL for methanol extracts of Echinops persicus and Vaccaria oxyodontha Boiss on S.aureus and E.coli at 31.25 mg/mL and 125 mg/mL,respectively.The highest diameter of inhibition zone caused by methanol extract of Cardamine uiliginosa was against S.aureus and E.coli with an inhibition zone of 23.9±0.29 mm and 20.4±0.43 mm,respectively.Compared with the antibiotics gentamicin and vancomycin,the Cardamine uliginosa showed a stronger inhibitory effect(P<0.05).Conclusion:Echinops persicus,Cardamine uliginosa,and Vaccaria oxyodontha Boiss have anti-bacterial properties.The anti-bacterial property of Cardamine uilignosa is higher than the other two herbal medicine.

Effect of auricular point sticking therapy on infantile amblyopia

Objective To compare the differences of effect on infantile amblyopia treated with auricular point sticking therapy and regular comprehensive therapy of western medicine.Methods Two hundreds and thirty cases of amblyopia patients were randomly divided into two groups,namely the group of auricular point sticking therapy,which had 120 cases（212 eyes）,and the group of comprehensive therapy,which had 110 cases（194 eyes）.The seed of Semen Vaccariae was applied at the ear point such as the eye,ear Shenmen,the liver,the spleen and the kidney.Regular comprehensive therapy of western medicine included covering method,vision corrected with glasses and fine exercise.Vision improvements of the two groups were observed with a three-year follow up.Results The effective rates of the auricular point sticking therapy group and comprehensive therapy group of ametropic amblyopia were 81.0%（64/79） and 52.2%（36/69）,respectively.The effective rates of anisometropic amblyopia of the two groups were 73.1%（49/67） and 47.7%（31/65）,respectively.And the effective rates of the two groups of strabismic amblyopia were 71.2%（47/66） and 45.0%（27/60）,respectively.The therapeutic effects of auricular point sticking therapy on all kinds of amblyopia were better than those of the comprehensive therapy（all P0.05）.Conclusion The effect of auricular point sticking therapy on infantile amblyopia is better than that of regular comprehensive therapy of western medicine,which can obviously improve the vision of the patients with easy manipulations.