JPT2 Affects Trophoblast Functions and Macrophage Polarization and Metabolism, and Acts as a Potential Therapeutic Target for Recurrent Spontaneous Abortion.

Recurrent spontaneous abortion (RSA) is a pregnancy-related condition with complex etiology. Trophoblast dysfunction and abnormal macrophage polarization and metabolism are associated with RSA; however, the underlying mechanisms remain unknown. Jupiter microtubule-associated homolog 2 (JPT2) is essential for calcium mobilization; however, its role in RSA remains unclear. In this study, it is found that the expression levels of JPT2, a nicotinic acid adenine dinucleotide phosphate-binding protein, are decreased in the villous tissues of patients with RSA and placental tissues of miscarried mice. Mechanistically, it is unexpectedly found that abnormal JPT2 expression regulates trophoblast function and thus involvement in RSA via c-Jun N-terminal kinase (JNK) signaling, but not via calcium mobilization. Specifically, on the one hand, JPT2 deficiency inhibits trophoblast adhesion, migration, and invasion by inhibiting the JNK/atypical chemokine receptor 3 axis. On the other hand, trophoblast JPT2 deficiency contributes to M1 macrophage polarization by promoting the accumulation of citrate and reactive oxygen species via inhibition of the JNK/interleukin-6 axis. Self-complementary adeno-associated virus 9-JPT2 treatment alleviates embryonic resorption in abortion-prone mice. In summary, this study reveals that JPT2 mediates the remodeling of the immune microenvironment at the maternal-fetal interface, suggesting its potential as a therapeutic target for RSA.

[Role of Puerariae Lobatae Radix in Gegen Decoction for treatment of primary dysmenorrhea].

This study aimed to explore the characteristic role of Puerariae Lobatae Radix(PLR) in Gegen Decoction for the treatment of primary dysmenorrhea(PD). Estrogen(E_2) was combined with oxytocin to establish a mouse model of PD. The mice were randomly divided into a normal group, a model group, a Gegen Decoction group, a PLR-free Gegen Decoction group, a PLR group, and a positive drug group(ibuprofen). Writhing response times and writhing incubation of mice in each group were tested by behavio-ral assessment, and the serum levels of prostaglandin F_(2α)(PGF_(2α)), prostaglandin E_2(PGE_2), E_2, and progesterone(PROG) were detected by ELISA kits. Western blot method was adopted to detect cyclooxygenase-2(COX-2) and estrogen receptor alpha(ER_α) expression levels in uterine tissues. Doppler ultrasound was employed to detect changes in uterine artery blood flow in mice, including peak systolic blood flow velocity(maximum velocity), end-diastolic velocity(minimum velocity), peak systolic blood flow velocity/end-diastolic velocity(S/D), pulsatility index(PI), and resistive index(RI). Histopathological changes in the uterus were detected by HE staining. Based on the oxytocin-induced isolated uterine contraction model, the effects of Gegen Decoction, PLR-free Gegen Decoction, and PLR on the amplitude, frequency, and activity of isolated uterine contraction were compared to investigate the role of PLR in Gegen Decoction for the treatment of PD. The results showed that compared with the Gegen Decoction group, the PLR-free Gegen Decoction improved the indicators of PD except for E_2 content, ER_α expression, and uterine artery blood flow. PLR could significantly down-regulate the serum content of E_2 and the protein expression of uterine ER_α, and improve the uterine artery blood flow. The data suggested that PLR, as the sovereign drug of Gegen Decoction, might function in Gegen Decoction for the treatment of PD by mediating E_(2 )and improving the uterine artery blood flow.

Widely Targeted Metabolomics Analysis of Different Parts of Salsola collina Pall.

Salsola collina Pall has a long history of being used as a traditional medicine to treat hypertension, headache, insomnia, constipation and vertigo. However, only a few biologically active substances have been identified from S. collina. Here, the shoots and roots of S. collina, namely L-Sc and R-Sc, were studied. The primary and secondary metabolites were investigated using ultrahigh-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS). A total of 637 putative metabolites were identified and these metabolites were mainly classified into ten different categories. Correlation analysis, hierarchical clustering analysis, principal component analysis and orthogonal partial least squares discriminant analysis of metabolites showed that the L-Sc samples could be clearly separated from the R-Sc samples. Differential accumulated metabolite analysis revealed that most of differential primary metabolites were significantly lower in the L-Sc than in the R-Sc. Conversely, the major differential secondary metabolites had higher levels in the L-Sc than in the R-Sc. Further analysis indicated that the flavonoids were the major putative antioxidant components and most of putative antioxidant components exhibited higher relative concentrations in the L-Sc than the R-Sc. These results improve our understanding of metabolite accumulation and provide a reference for the study of medicinal value in S. collina.

Spasmolytic Mechanism of Aqueous Licorice Extract on Oxytocin-Induced Uterine Contraction through Inhibiting the Phosphorylation of Heat Shock Protein 27.

Licorice derived from the roots and rhizomes of Glycyrrhiza uralensis Fisch. (Fabaceae), is one of the most widely-used traditional herbal medicines in China. It has been reported to possess significant analgesic activity for treating spastic pain. The aim of this study is to investigate the spasmolytic molecular mechanism of licorice on oxytocin-induced uterine contractions and predict the relevant bioactive constituents in the aqueous extract. The aqueous extraction from licorice inhibited the amplitude and frequency of uterine contraction in a concentration-dependent manner. A morphological examination showed that myometrial smooth muscle cells of oxytocin-stimulated group were oval-shaped and arranged irregularly, while those with a single centrally located nucleus of control and licorice-treated groups were fusiform and arranged orderly. The percentage of phosphorylation of HSP27 at Ser-15 residue increased up to 50.33% at 60 min after oxytocin stimulation. Furthermore, this increase was significantly suppressed by licorice treatment at the concentration of 0.2 and 0.4 mg/mL. Colocalization between HSP27 and α-SMA was observed in the myometrial tissues, especially along the actin bundles in the oxytocin-stimulated group. On the contrary, the colocalization was no longer shown after treatment with licorice. Additionally, employing ChemGPS-NP provided support for a preliminary assignment of liquiritigenin and isoliquiritigenin as protein kinase C (PKC) inhibitors in addition to liquiritigenin, isoliquiritigenin, liquiritin and isoliquiritin as MAPK-activated protein kinase 2 (MK2) inhibitors. These assigned compounds were docked with corresponding crystal structures of respective proteins with negative and low binding energy, which indicated a high affinity and tight binding capacity for the active site of the kinases. These results suggest that licorice exerts its spasmolytic effect through inhibiting the phosphorylation of HSP27 to alter the interaction between HSP27 and actin. Furthermore, our results provide support for the prediction that potential bioactive constituents from aqueous licorice extract inhibit the relevant up-stream kinases that phosphorylate HSP27.

Bioactive prenylated xanthones from the stems of Cratoxylum cochinchinense.

Cochinchinones M-U (1-9), together with 12 known compounds (10-21), were isolated from the stems of Cratoxylum cochinchinense (Lour.) Blume. Their structures were determined on the basis of extensive spectroscopic data analyses. In addition, their retinoid X receptor-α transcriptional activities were evaluated using an in vitro assay.

Fluoride adsorption properties of three modified forms of activated alumina in drinking water.

The study describes the removal of fluoride from drinking water using activated alumina (AA). AA was modified with H2SO4, FeCl3 and a combination of the two to enhance fluoride adsorption. The AA adsorbents were characterized using Brunauer-Emmett-Teller surface area analysis and X-ray fluorescence. The maximum adsorption capacity of H2SO4- and FeCl3-modified AA adsorbents was 4.98 mg/g, which is 3.4 times higher compared with that of normal AA. The results showed that the surface area of AA increased when modified with H2SO4. AA modified with FeCl3 enhanced fluoride adsorption ability through ion-exchange between chlorine ions and fluoride ions. The fluoride adsorption properties of AA modified with both H2SO4 and FeCl3 were consistent with the Langmuir model. The fluoride adsorption kinetics of the adsorbents were well described by the pseudo-second-order kinetic model.

[Lignans isolated from stems of Sambucus williamsii and their proliferation effects on UMR106 cells].

The present study aims to investigate the lignan constituents from Sambucus williamsii and their proliferation effects on osteoblast-like UMR106 cells. Seven compounds were isolated and purified by macroporous resin D101, silica gel, Sephadex LH-20, Toyopearl HW-40, ODS column chromatographies and Preparative HPLC(C-18). Their structures were elucidated by spectroscopic methods as threo-guaiacylglycerol-beta-0-4'-conifery ether (1), lirioresinol A (2), 1-hydroxypinoresinol (3), 5-methoxybalanophonin (4), balanophonin (5), 5-methoxy-trans-dihydrodehydrodiconiferyl alcohol (6), and p-hydroxybenzaldehyde (7). Compounds 3-7 were obtained from this genus for the first time. The proliferation effects of all isolated compounds on osteoblast-like UMR106 cells were determined. Compounds 1-7 (1 x 10(-12)-1 x 10(-7) mol x L(-1)) increased UMR106 cell proliferation to some extent.

A new lignan glycoside from Forsythia suspensa.

Phytochemical investigation on Forsythia suspensa (Thunb.) Vahl afforded ten compounds, including five lignan glycosides and five phenylethanoid glycosides. The compounds were isolated by using HP-20 macroporous resin, silica gel, octadecyl silica gel (ODS), size exclusion chromatography resin HW-40 chromatography, and preparative HPLC. The structures were established through application of extensive spectroscopic methods, including ESI-MS, 1D-and 2D-NMR spectroscopy. They were identified as forsythialanside E (1), 8'-hydroxypinoresinol-4'-O-ß-D-glucoside (2), 8'-hydroxypinoresinol (3), lariciresinol-4'-O-ß-D-glucoside (4), lariciresinol-4-O-ß-D-glucoside (5), forsythoside H (6), forsythoside I (7), forsythoside F (8), plantainoside B (9), and plantainoside A (10). Compound 1 was a new lignan glycoside.

New ursane-type triterpenoid saponins from the stem bark of Schefflera heptaphylla.

Phytochemical investigation on the stem bark of Schefflera heptaphylla led to the isolation of five new ursane-type triterpenoid saponins (1-5). Their structures were determined on the basis of spectroscopic and chemical methods. It is noteworthy in this study that the genins of all compounds are reported for the first time. All compounds isolated from this plant were evaluated for their inhibitory activities on lipopolysaccharide-induced nitric oxide production in RAW264.7 cells, and compounds 2 and 5 showed weak anti-inflammatory activities under their non-cytotoxic concentrations.

Triterpenoid saponins from the stem barks of Schefflera heptaphylla.

Nine new triterpenoid saponins, including four ursane-type triterpenoid saponins named heptursosides A-D (1-4), four oleanane-type triterpenoid saponins named heptoleosides A-D (5-8), and one dammarane-type triterpenoid saponin, heptdamoside A (9), along with two known saponins, asiaticoside D (10) and scheffoleoside B (11), were isolated from the stem barks of Schefflera heptaphylla. Their structures were determined on the basis of spectroscopic analysis and chemical methods. It is noteworthy in this study that the aglycone of 1-6 is reported for the first time, and to the best of our knowledge, this is the first report for the presence of the tetracyclic triterpenoid saponin from Schefflera. All the saponins were evaluated for their inhibitory effects on lipopolysaccharide-induced nitric oxide production in RAW264.7 cells, and 2, 6, 7, and 10 showed anti-inflammatory activities under their noncytotoxic concentrations.

Guava leaf extracts promote glucose metabolism in SHRSP.Z-Leprfa/Izm rats by improving insulin resistance in skeletal muscle.

BACKGROUND: Metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM) have been associated with insulin-resistance; however, the effective therapies in improving insulin sensitivity are limited. This study is aimed at investigating the effect of Guava Leaf (GL) extracts on glucose tolerance and insulin resistance in SHRSP.Z-Leprfa/Izm rats (SHRSP/ZF), a model of spontaneously metabolic syndrome. METHODS: Male rats at 7 weeks of age were administered with vehicle water or treated by gavage with 2 g/kg GL extracts daily for six weeks, and their body weights, water and food consumption, glucose tolerance, and insulin resistance were measured. RESULTS: Compared with the controls, treatment with GL extracts did not modulate the amounts of water and food consumption, but significantly reduced the body weights at six weeks post treatment. Treatment with GL extracts did not alter the levels of fasting plasma glucose and insulin, but significantly reduced the levels of plasma glucose at 60 and 120 min post glucose challenge, also reduced the values of AUC and quantitative insulin sensitivity check index (QUICKI) at 42 days post treatment. Furthermore, treatment with GL extracts promoted IRS-1, AKT, PI3Kp85 expression, then IRS-1, AMKP, and AKT308, but not AKT473, phosphorylation, accompanied by increasing the ratios of membrane to total Glut 4 expression and adiponectin receptor 1 transcription in the skeletal muscles. CONCLUSIONS: These data indicated that GL extracts improved glucose metabolism and insulin sensitivity in the skeletal muscles of rats by modulating the insulin-related signaling.

Effects of estrogen and phytoestrogens on endometrial leakage in ovariectomized rats and the related mechanisms.

Phytoestrogens, a group of plant-derived non-steroidal compounds that can behave as estrogens by binding to estrogen receptors, have drawn great attention for their potentially beneficial effects on human health. However, there are few studies investigating the potential side effects of phytoestrogens on the reproductive system. The present study was to elucidate the effects of 17ß-estradiol (E2), progesterone (P4), and phytoestrogens genistein (Gen), resveratrol (Res), and phloretin (Phl) on eosinophilic infiltration of the ovariectomized rat uterus and endometrial vascular permeability, and to analyze the underlying mechanisms. The ovariectomized rats received daily subcutaneous injections of E2, E2+P4, P4, Gen, Res, Phl, or an equivalent volume of vehicle for 21 days, and sham-operated animals (Sham rats) were used as the controls. Hematoxylin-eosin staining revealed a marked increase in uterine eosinophilic infiltrations in ovariectomized rats treated with E2, E2+P4 or P4, which was associated with increased expression of vascular endothelial growth factor (VEGF), nuclear factor-κB (NF-κB), and tumor necrosis factor-α (TNF-α) proteins as determined by immunohistochemical and Western blot analysis. However, all three phytoestrogens had no markedly effect on the uterine eosinophilic infiltration and the expressions of VEGF, NF-κB, and TNF-α in the uterus of ovariectomized rats. Our data demonstrate that E2 alone or in combination with P4 increases uterine eosinophilic infiltration which is related with vascular hyperpermeability caused by VEGF, NF-κB and TNF-α, whereas phytoestrogens Gen, Res, and Phl, have no such an effect.

Targeting truncated retinoid X receptor-α by CF31 induces TNF-α-dependent apoptosis.

A truncated version of retinoid X receptor-α, tRXR-α, promotes cancer cell survival by activating the phosphoinositide 3-kinase (PI3K)/AKT pathway. However, targeting the tRXR-α-mediated survival pathway for cancer treatment remains to be explored. We report here our identification of a new natural product molecule, CF31, a xanthone isolated from Cratoxylum formosum ssp. pruniflorum, and the biologic evaluation of its regulation of the tRXR-α-mediated PI3K/AKT pathway. CF31 binds RXR-α and its binding results in inhibition of RXR-α transactivation. Through RXR-α mutational analysis and computational studies, we show that Arg316 of RXR-α, known to form salt bridges with certain RXR-α ligands, such as 9-cis-retinoic acid (9-cis-RA), is not required for the antagonist effect of CF31, showing a distinct binding mode. Evaluation of several CF31 analogs suggests that the antagonist effect is mainly attributed to an interference with Leu451 of helix H12 in RXR-α. CF31 is a potent inhibitor of AKT activation in various cancer cell lines. When combined with TNF-α, it suppresses TNF-α activation of AKT by inhibiting TNF-α-induced tRXR-α interaction with the p85α regulatory subunit of PI3K. CF31 inhibition of TNF-α activation of AKT also results in TNF-α-dependent activation of caspase-8 and apoptosis. Together, our results show that CF31 is an effective converter of TNF-α signaling from survival to death by targeting tRXR-α in a unique mode and suggest that identification of a natural product that targets an RXR-mediated cell survival pathway that regulates PI3K/AKT may offer a new therapeutic strategy to kill cancer cells.

[Study on the HPLC fingerprint and simultaneous determination of three triterpene glycosides in Schefflera heptaphylla].

OBJECTIVE: To establish the HPLC fingerprint and simultaneously determination three triterpene glycosides in Schefflera heptaphylla. METHODS: The separation was carried out on an Agilent Zorbax SB-C18 (4.6 mm x 250 mm, 5 microm) column eluted with methanol--0.1% glacial acetic acid as the mobile phase, and samples were detected by an evaporative light scattering detector (ELSD). The similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine (Version 2004 A) was used in data analysis. RESULTS: Schefflera heptaphylla from different regions were similar and the separation of three triterpene glycosides was good. CONCLUSION: This method is simple and reliable. The HPLC fingerprint and contents of three kinds of triterpene glycosides of Schefflera heptaphylla can be used for their quality control.

A new phenylpropanoid glucoside from the aerial parts of Lygodium japonicum.

A new compound, 4-O-caffeoyl-D-glucopyranose (1), and a new natural product, 3-O-caffeoyl-D-glucopyranose (2), together with six known compounds (3-8) were isolated from the aerial parts of Lygodium japonicum. Their structures were elucidated on the basis of extensive spectroscopic data analyses. The oxygen radical absorbance capacity assay was applied to evaluate their antioxidative capacities in vitro, which revealed that 1-8 showed strong antioxidative properties.

A new ursane-type triterpenoid from Schefflera heptaphylla (L.) Frodin.

A new ursane-type triterpenoid (1), together with 15 known compounds (2-16), was isolated from the barks of Schefflera heptaphylla (L.) Frodin. The structure of the new compound was determined on the basis of extensive spectroscopic data including IR, HR-ESI-MS, 1D and 2D NMR, and further confirmed by single-crystal X-ray diffraction. Compounds 2-6 were isolated from Schefflera genus for the first time.

Xanthone and benzophenone glycosides from the stems of Cratoxylum formosum ssp. pruniflorum.

Two new xanthone glycosides, namely pruniflorosides A and B (1, 2), a new benzophenone glycoside, prunifloroside C (3), and a new xanthone, pruniflorone S (4) were isolated from the stems of Cratoxylum formosum ssp. pruniflorum, along with six known xanthones (5-10). Their structures were determined on the basis of extensive spectroscopic analysis. In addition, their retinoid X receptor α (RXRα) transcriptional activities were evaluated in vitro.

[Study on the chemical constituents of roots and stems of Nardostachys chinensis].

OBJECTIVE: To study the chemical constituents of roots and stems of Nardostachys chinensis. METHODS: The chemical constituents were isolated and purified by column chromatography including silica gel, Diaion HP-20, Sephadex LH-20, ODS, and HPLC. Their structures were elucidated on the basis of spectral data analyses. RESULTS: Six compounds were isolated and identified as 8-hydroxypinoresinol-4-O-beta-D-glucopyranoside (1), 8-hydroxypinoresinol-4'-O-beta-D-glucopyranoside (2), 8-hydroxypinoresinol-4-O-(beta-D-glucopyranosyl) 4'-O-beta-D-glucopyranoside (3), cycloolivil 6-O-beta-D-glucopyranoside(4), ferulic acid (5), chlorogenic acid methyl ester (6). CONCLUSION: All the compounds are isolated from this genus for the first time.

Regulation of expression and activity of selenoenzymes by different forms and concentrations of selenium in primary cultured chicken hepatocytes.

The expression and activity of selenoenzymes are regulated by Se. In the present study, the effects of different forms and concentrations of Se on the regulation of glutathione peroxidase (GPx) activity and phospholipid hydroperoxide GPx (GPx4) and type I deiodinase (D1) mRNA levels in chicken hepatocytes were evaluated. Primary cultured chicken hepatocyte monolayers derived from male White Leghorn chickens (aged 30-40 d) were incubated for 24 h with 0 (control), 0.5, 1, 1.5, 2, 3, 4 or 5 µmol/l of Se supplied as dl-selenomethionine (Se-Met), κ-selenocarrageenan (Se-Car) or sodium selenite (Na2SeO3). Compared with the control, Se significantly increased GPx activity in all the hepatocytes, but the activity was not increased in the hepatocytes treated with 5 µmol/l of Na2SeO3, with maximal effects being observed at 2 µmol/l of Se-Met or Se-Car and at 1.5 µmol/l of Na2SeO3, respectively. Significant decreases in GPx4 mRNA levels were observed in all the hepatocytes treated with Se (v. control). The D1 mRNA levels were significantly increased in all the groups treated with Se (v. control), with maximal effects being observed at 1.5 µmol/l of Se-Met and at 0.5 µmol/l of Se-Car or Na2SeO3, respectively. Se-Met at doses of 1.5-5 µmol/l had a greater effect on D1 mRNA than Se-Car and Na2SeO3 at equivalent doses. After resulting in a maximal effect, higher Se supplementation led to a dose-dependent reduction in GPx activity and D1 mRNA levels in all the hepatocytes treated with Se. These results suggest that in chicken hepatocytes, the regulations of GPx and D1 by different forms and concentrations of Se vary.

Bioactive xanthones from the stems of Cratoxylum formosum ssp. pruniflorum.

Six new compounds, pruniflorones M-R (1-6), together with 19 known compounds (7-25) were isolated from the stems of Cratoxylum formosum ssp. pruniflorum. The structures of the new compounds were established on the basis of extensive spectroscopic data interpretation. In addition, their RXRalpha transcriptional activities were evaluated using an in vitro assay.