Fibulin2: a negative regulator of BMSC osteogenic differentiation in infected bone fracture healing.

Bone fracture remains a common occurrence, with a population-weighted incidence of approximately 3.21 per 1000. In addition, approximately 2% to 50% of patients with skeletal fractures will develop an infection, one of the causes of disordered bone healing. Dysfunction of bone marrow mesenchymal stem cells (BMSCs) plays a key role in disordered bone repair. However, the specific mechanisms underlying BMSC dysfunction caused by bone infection are largely unknown. In this study, we discovered that Fibulin2 expression was upregulated in infected bone tissues and that BMSCs were the source of infection-induced Fibulin2. Importantly, Fibulin2 knockout accelerated mineralized bone formation during skeletal development and inhibited inflammatory bone resorption. We demonstrated that Fibulin2 suppressed BMSC osteogenic differentiation by binding to Notch2 and inactivating the Notch2 signaling pathway. Moreover, Fibulin2 knockdown restored Notch2 pathway activation and promoted BMSC osteogenesis; these outcomes were abolished by DAPT, a Notch inhibitor. Furthermore, transplanted Fibulin2 knockdown BMSCs displayed better bone repair potential in vivo. Altogether, Fibulin2 is a negative regulator of BMSC osteogenic differentiation that inhibits osteogenesis by inactivating the Notch2 signaling pathway in infected bone.

Effect of Alpinia officinarum Hance alcohol extracts on primary dysmenorrheal.

OBJECTIVE: To study the effect of Alpinia officinarum Hance (A. officinarum) 80% alcohol extract on the primary dysmenorrhea. METHODS: A. officinarum 80% alcohol extract were enriched by macroporous adsorption resins. Female mice of primary dysmenorrhea model were established by oxytocin induction; the effects of A. officinarum 80% alcohol extract on primary dysmenorrhea were observed by body twist method; and the homogenate level of prostaglandin F2α (PGF2α), prostaglandin E2 (PGE2) and Ca(2+) in the uterus were observed in oxytocin-induced female mice. RESULTS: The writhing frequency of primary dysmenorrhea mice was significantly decreased after treatment of A. officinarum 80% alcohol extract and the level of PGF2α, PGE2 and Ca(2+) in mice uterus was significantly decreased (P < 0.05, P < 0.01) in groups of mice treated with middle and high dosage of A. officinarum 80% alcohol extract compared with that of model group. CONCLUSIONS: These findings suggest that A. Officinarum 80% alcohol extract can significantly relieve primary dysmenorrhea.

Differential systemic exposure to galangin after oral and intravenous administration to rats.

BACKGROUND: Galangin (3,5,7-trihydroxyflavone) is present in high concentrations in herbal medicine such as Alpinia officinarum Hance. Galangin shows multifaceted in vitro and in vivo biological activities. The number and position of hydroxyl groups in this molecule play an important role in these biological activities. However, these hydroxyl groups undergo glucuronidation and sulfation in in vitro assay system. However, the systemic exposure to galangin after dosing in animals and/or humans remains largely unknown. Thus it is not clear whether the galangin exists in the body at concentrations high enough for the biological effects. Furthermore, the metabolite identification and the corresponding plasma pharmacokinetics need to be characterized. RESULTS: Two LC-MS/MS methods were developed and validated and successfully applied to analyze the parent drug molecules and aglycones liberated from plasma samples via ß-glucuronidase hydrolysis. Our major findings were as follows: (1) The routes of administration showed significant influences on the systemic exposure of galangin and its metabolites. (2) Galangin was preferentially glucuronidated after p.o. dosing but sulfated after i.v. medication. (3) Kaempferol conjugates were detected demonstrating that oxidation reaction occurred; however, both glucuronidation and sulfation were more efficient. (4) Oral bioavailability of free parent galangin was very low. CONCLUSIONS: Systemic exposure to galangin and its metabolites was different in rat plasma between oral and intravenous administration. Further research is needed to characterize the structures of galangin conjugates and to evaluate the biological activities of these metabolites. Graphical abstractGalangin was preferentially glucuronidated after p.o. dosing but sulfated after i.v. medication.

Three new compounds isolated from the seeds of Vaccaria segetalis.

Two new capsaicin analogs, N-(3-methoxy-4-hydroxyphenethyl)-tetracosanamide (1) and N-(3,4-dihydroxyphenethyl)-tetracosanamide (2), along with one new flavonoidal glycoside pinnatifin E (3) were isolated from the ethanolic extract of the seeds of Vaccaria segetalis. Their structures were elucidated on the basis of spectroscopic methods including 1D, 2D NMR, MS, and other spectroscopic techniques, as well as by comparison with the relevant literatures. All compounds were evaluated for their coagulation Factor Xa inhibition activities.

Quantitative evaluation of Radix Paeoniae Alba sulfur-fumigated with different durations and purchased from herbal markets: simultaneous determination of twelve components belonging to three chemical types by improved high performance liquid chromatography-diode array detector.

In this study, a improved high performance liquid chromatography-diode array detector (HPLC-DAD) method for simultaneous quantification of twelve major components belonging to three chemical types was developed and validated, and was applied to quantitatively compare the quality of Radix Paeoniae Alba (RPA) sulfur-fumigated with different durations and purchased from commercial herbal markets. The contents of paeoniflorin, benzoylpaeoniflorin, oxypaeoniflorin, benzoic acid and paeonol decreased whereas that of paeoniflorin sulfonate increased in RPA with the extending of sulfur-fumigation duration. Different levels of paeoniflorin sulfonate were determined in ten of seventeen commercial RPA samples, indicating that these ten samples may be sulfur-fumigated with different durations. Moreover, the relative standard deviation of the contents of each component was higher in the commercial sulfur-fumigated RPA samples than that in commercial non-fumigated RPA samples, and the percentage of the total average content of monoterpene glycosides in the determined analytes was higher in the decoctions of commercial sulfur-fumigated RPA than that in commercial non-fumigated RPA samples. All these results suggested that the established method was precise, accurate and sensitive enough for the global quality evaluation of sulfur-fumigated RPA, and sulfur-fumigation can not only change the proportions of bioactive components, but also cause the reduction of the quality consistency of both raw materials and aqueous decoctions of RPA.

Matrix metalloproteinase-9 was involved in the immuno-modulatory defect of mesenchymal stem cell from chronic myeloid leukemia patients.

BACKGROUND: Overwhelming evidences on chronic myeloid leukemia (CML) indicate that patients harbor quiescent CML stem cells that are responsible for blast crisis. While the hematopoietic stem cell (HSC) origin of CML was first suggested over 30 years ago, recently CML-initiating cells beyond HSCs are also being investigated. METHODS: We have previously isolated fetal liver kinase-1-positive (Flk1(+)) cells carrying the BCR/ABL fusion gene from the bone marrow of Ph(+) patients with hemangioblast property. In this study, we isolated CML patient-derived Flk1(+)CD31(-)CD34(-) mesenchymal stem cells (MSCs) and detected their biological characteristics and immunological regulation using fluorescence in situ hybridization (FISH) analysis, fluorescence activated cell sorting (FACS), enzyme-linked immunoadsorbent assay, mixed lymphocyte reaction assays; then we compared these characters with those of the healthy donors. RESULTS: CML patient-derived Flk1(+)CD31(-)CD34(-) MSCs had normal morphology, phenotype and karyotype while appeared impaired in immuno-modulatory function. The capacity of patient Flk1(+)CD31(-)CD34(-) MSCs to inhibit T lymphocyte activation and proliferation was impaired in vitro. CONCLUSIONS: CML patient-derived MSCs have impaired immuno-modulatory functions, suggesting that the dysregulation of hematopoiesis and immune response may originate from MSCs rather than hematopoietic stem cells (HSCs). MSCs might be a potential target for developing efficacious treatment for CML.

A new phenolic glycoside from prunella vulgaris.

A new phenolic glycoside was isolated from the spikes of Prunella vulgaris. Its structure was elucidated as gentisic acid 5-O-beta-D-(6'-salicylyl)-glucopyranoside by spectroscopic evidence and chemical analysis.

Caudatin-2,6-dideoxy-3-O-methy-ß-D-cymaropyranoside 1 induced apoptosis through caspase 3-dependent pathway in human hepatoma cell line SMMC7721.

Caudatin-2,6-dideoxy-3-O-methy-ß-D-cymaropyranoside (CDMC), the C-21 steroidal glycoside recently extracted from the traditional Chinese medicinal plant, the root of Cynanchum auriculatum Royle ex Wight (Asclepiadaceae), has been shown to possess potent antitumor properties. However, the bioactivities of CDMC are still largely unknown, especially the antitumor effect and its mechanism. This study investigated the CDMC antitumor effects on human hepatoma cell line SMMC7721 cells by analysis of cell viability, cell cycle phases and apoptosis. The results showed that CDMC inhibited the growth of SMMC7721 cells in a time- and dose-dependent manner and resulted in cell cycle arrest in G(0)/G(1) phase. Furthermore, CDMC induced SMMC7721 cell apoptosis rather than necrosis through caspase 3 activation, and a caspase 3 inhibitor, Ac-DEVD-CHO, could attenuate the apoptosis induced by CDMC. The results suggested that the anticancer activity of CDMC could be attributed partially to its inhibition of cell proliferation and induction of apoptosis associated with caspase 3 activation.

Four anthraquinones from Hedyotis diffusa.

A new anthraquinone has been isolated from the 95% EtOH extract of Hedyotis diffusa and characterized as 2-hydroxy-3-methoxy-6-methyl-9,10-anthraquinone (1) by extensive spectral analysis. The known compounds isolated for the first time from this plant have been identified as 2-hydroxy-3-methoxy-7-methyl-9,10-anthraquinone (2), 2-hydroxy-6-methylanthraquinone (3), and 1,3-dimethoxy-2-hydroxy-9,10-anthraquinone (4).

Antitumor activity of C-21 steroidal glycosides from Cynanchum auriculatum Royle ex Wight.

The antitumor activities of six C-21 steroidal glycosides isolated from the root tuber of Cynanchum auriculatum Royle ex Wight were performed according to a microculture tetrazolium (MTT) method on human tumor cell lines SMMC-7721, MCF-7 and Hela. Of these compounds, caudatin-2,6-dideoxy-3-O-methy-ß-D-cymaropyranoside and caudatin were found to be of the highest effects against human tumor cell line SMMC-7721 with IC(50) values of 13.49 and 24.95 µM, respectively. Then the in vivo assay further showed that caudatin-2,6-dideoxy-3-O-methy-ß-D-cymaropyranoside and caudatin significantly inhibited the growth of transplantable H(22) tumors in mice.

[Chemical constituents from Hedyotis diffusa].

OBJECTIVE: To investigate the chemical constituents from Hedyotis diffusa. METHOD: The compounds were isolated and purified by various chromatographic techniques and identified by their physicochemical properties and spectral data. RESULT: Eight compounds were isolated and identified as octadecyl (E)-p-coumarate (1), p-E-methoxy-cinnamic acid (2), ferulic acid (3), scopoletin (4), succinic acid (5), aurantiamide acetate (6), rubiadin (7), robustaquinone D (8). CONCLUSION: Compounds 1-8 were obtained from genus Hedyotis for the first time.

A new alkaloid from Salsola collina.

Salsola collina is widely distributed in droughty and semi-droughty area, which is used as a kind of folk remedy in traditional Chinese medicine for treatment of hypertension. The study is on the chemical constituents of this herb from its aerial parts to obtain its active constituents. Dried and crushed aerial parts of this herb were extracted three times with 95% EtOH at reflux. The ethanol extracts were combined and concentrated under reduced pressure at 70 degrees C to yield residue, which was suspended in water and successively partitioned with light petroleum, chloroform and n-butanol. The chloroform and n-butanol fractions were treated by various chromatographic techniques, such as silica gel, C18 reversed-phase silica gel and macroporous resin column chromatography. Compounds were elucidated by their physicochemical properties and spectroscopic analysis. In the course of our study on searching biological active components from this herb, a new alkaloid together with three known alkaloids were isolated and identified as N-transferuloyl-3-methyldopamine (1), 3-[4-(beta-D-glucopyranosyloxy)-3-methoxyphenyl]-N-[2-(4-hydroxyl-3-methoxyphenyl) ethyl]-2-propenamide (2), salsoline A (3), salsoline B (4). Compound 4 is a new compound and named as salsoline B, while compound 2 was obtained in Salsola collina for the first time.

[Chemical constituents from roots Pseudostellaria heterophylla].

OBJECTIVE: To study the chemical constituents of Pseudostellaria heterophylla. METHOD: The compounds were isolated by silica gel and Sephadex LH-20 column chromatography and purified by recrystallization. Their structures were elucidated on the basis of physicochemical properties and spectral analysis. RESULT: Seven compounds were identified as 2-minalin (1) , 3-furfuryl pyrrole-2-carboxylate (2), ursolic acid (3), acacetin (4), luteolin (5), acacetin 7-O-beta-D-glucopyranosyl (6-->1)-alpha-L-rhamnopyranoside (6). CONCLUSION: Among them, compounds 3-6 were isolated from the plant for the first time.

[Studies on chemical constituents of Prunella vulgaris].

OBJECTIVE: To study the chemical constituents of Prunella vulgaris. METHOD: To separate the constituents of P. vulgaris by using various kinds of chromatography and identify their structures on the basis of spectral analysis. RESULT: Seven compounds were isolated from the spikes of P. vulgaris. Their structures were established as autantiamide acetate (1), rhein (2), tanshinone I (3), danshensu (4), stigmast-7, 22-dien-3-one (5), 3, 4, alpha-trihydroxy-methyl phenylpropionate (6), butyl rosmarinate (7). CONCLUSION: Compounds 1-4 were isolated from this genus for the first time.

[Studies on chemical constituents of Salsola collina].

OBJECTIVE: To study the chemical constituents of Salsola collina. METHOD: The EtOH extract from the whole plant of S. collina were isolated and purified by column chromatography. These compounds were identified by their physical properties and spectroscopic data. RESULT: Eleven compounds were isolated and identified as ferulic acid (1), p-coumaric acid (2), salicylic acid (3), tricin (4), selagin (5), acanthoside D (6), tricin -7-O-beta-D-glucopyranoside (7), tricin-4'-O-beta-D-apioside (8), isorhamnetin-7-O-beta-D-glucopyranoside (9), isorhamnetin-3-O-beta-D-glucopyranoside (10), isorhamnetin-3-O-alpha-L-arabinopyranosyl (1-->6) -beta-D-glucopyranoside (11). CONCLUSION: wherein compound 1, 2, 3, 5, 6 and 9 were isolated from the Salsola for the first time.

[Studies on chemical constituents in root tuber of Cynanchum auriculatum].

OBJECTIVE: To study chemical constituents from Cynanchum auriculatum. METHOD: The chemical components were isolated and purified by silca gel, sephedex-LH-20 and ODS column chromatography. The chemical structures were elucidated on the basis of physicochemical properties and spectral data. RESULT: Thirteen compounds were isolated and identified as: caudatin (1), metaplexigenin (2), cynauricuoside A (3), succinic acid (4), azelaic acid (5), wilforibiose (6), sucrose (7), 1-O-hexadecanolenin (8), beta-amyrin acetate (9), cynanchone A (10), acetylquinol (11), beta-sitosterol (12), daucosterol (13). CONCLUSION: Compounds 4-9 were obtained from this plant for the first time.

[Isolation and characterization of phenylethanoid glycosides from Clerodendron bungei].

AIM: To study the chemical constituents from Clerodendron bungei Steud. METHODS: The compounds were isolated and purified by various chromatographic techniques and identified by their physicochemical properties and spectral data. RESULTS: Ten phenylethanoid glycosides were isolated and identified as clerodendronoside (1), acteoside (2), isoacteoside (3), cistanoside C (4), jionoside C (5), leucosceptoside A (6), cistanoside D (7), campneoside I (8), campneoside II (9), cistanoside F (10). CONCLUSION: Compound 1 is a new phenylethanoid glycoside, while compounds 4-10 are obtained from this plant for the first time.