Jinfeng pills ameliorate premature ovarian insufficiency induced by cyclophosphamide in rats and correlate to modulating IL-17A/IL-6 axis and MEK/ERK signals.

ETHNOPHARMACOLOGICAL RELEVANCE: Jinfeng Pill (JFP) is a classical Chinese medicine formula and composed of 9 herbs, including Epimedium brevicornu Maxim (Yinyanghuo), Cervus elaphus Linnaeus (Lurong), Panax ginseng C.A.Mey. (Renshen), Equus asinus (EJiao), Ligustrum lucidum W.T.Aiton (Nvzhenzi), Reynoutria multiflora (Thunb.) Moldenke (Heshouwu), Curculigo orchioides Gaertn (Xianmao), Neolitsea cassia (L.) Kosterm. (Rougui) and Leonurus japonicus Houtt. (Yimucao). The formula is clinically used to regulate menstrual cycle and alleviate polycystic ovarian syndrome due to its capabilities of ovulation induction. It is therefore presumed that JFP could be used for the therapy of premature ovarian insufficiency (POI) but the assumed efficacy has not been fully substantiated in experiment. AIM OF STUDY: To evaluate the effectiveness of JFP on cyclophosphamide (CTX)-induced POI and preliminarily explore its potential mechanisms of action. MATERIAL AND METHODS: An experimental rat model of POI was established by using CTX induction to assess the efficacy of JFP. The potential targets of action for JFP alleviating POI were predicted by the combination of network pharmacology and transcriptomics and finally validating by RT-qPCR and Western blot. RESULTS: JFP alleviated the damages of ovarian tissue induced by CTX in the rat model of POI via significantly decreasing serum levels of FSH and LH and the ratio of FSH/LH and increasing the levels of E2 and AMH, accompanied with promoting ovarian folliculogenesis and follicle maturity and reversing the depletion of follicle pool. With the analysis of network pharmacology, pathways in cancer, proteoglycans in cancer, PI3K-AKT, TNF and FoxO signaling pathways were predicted to be influenced by JFP. The results of RNA-seq further revealed that IL-17 signaling pathway was the most important pathway regulated by both CTX and JFP, following by transcriptional misregulation in cancer and proteoglycans in cancer. Combining the two analytical methods, JFP likely targeted genes associated with immune regulation, including COX-2, HSP90AA1, FOS, MMP3 and MAPK11 and pathways, including IL-17,Th17 cell differentiation and TNF signaling pathway. Finally, JFP was validated to regulate the mRNA expression of FOS, FOSB, FOSL1, MMP3, MMP13 and COX-2 and decrease the release of IL-17A and the protein expression of IL-6 and suppress the phosphorylation of MEK1/2 and ERK1/2 in CTX induced POI rats. CONCLUSION: Jinfeng Pill is effective to ameliorate the symptoms of POI induced by CTX in the model of rats and its action is likely associated with suppressing IL-17A/IL-6 axis and the activity of MEK1/2-ERK1/2 signaling.

Metformin Enhances Nomegestrol Acetate Suppressing Growth of Endometrial Cancer Cells and May Correlate to Downregulating mTOR Activity In Vitro and In Vivo.

This study investigated the effect of a novel progestin and its combination with metformin on the growth of endometrial cancer (EC) cells. Inhibitory effects of four progestins, including nomegestrol acetate (NOMAC), medroxyprogesterone acetate, levonorgestrel, and cyproterone acetate, were evaluated in RL95-2, HEC-1A, and KLE cells using cell counting kit-8 assay. Flow cytometry was performed to detect cell cycle and apoptosis. The activity of Akt (protein kinase B), mTOR (mammalian target of rapamycin) and its downstream substrates 4EBP1 (4E-binding protein 1) and eIF4G (Eukaryotic translation initiation factor 4G) were assayed by Western blotting. Nude mice were used to assess antitumor effects in vivo. NOMAC inhibited the growth of RL95-2 and HEC-1A cells, accompanied by arresting the cell cycle at G0/G1 phase, inducing apoptosis, and markedly down-regulating the level of phosphorylated mTOR/4EBP1/eIF4G in both cell lines (p < 0.05). Metformin significantly increased the inhibitory effect of and apoptosis induced by NOMAC and strengthened the depressive effect of NOMAC on activity of mTOR and its downstream substrates, compared to their treatment alone (p < 0.05). In xenograft tumor tissues, metformin (100 mg/kg) enhanced the suppressive effect of NOMAC (100 mg/kg) on mTOR signaling and increased the average concentration of NOMAC by nearly 1.6 times compared to NOMAC treatment alone. Taken together, NOMAC suppressing the growth of EC cells likely correlates to down-regulating the activity of the mTOR pathway and metformin could strengthen this effect. Our findings open a new window for the selection of progestins in hormone therapy of EC.

Identification of New Epididymal Luminal Fluid Proteins Involved in Sperm Maturation in Infertile Rats Treated by Dutasteride Using iTRAQ.

BACKGROUND: Spermatozoa become mature and acquire fertilizing capacity during their passage through the epididymal lumen. In this study, we identified new epididymal luminal fluid proteins involved in sperm maturation in infertile rats by dutasteride, a dual 5α-reductase inhibitor, in order to provide potential epididymal targets for new contraceptives and infertility treatment. METHODS: Male rats were treated with dutasteride for 28 consecutive days. We observed the protein expression profiles in the epididymal luminal fluids in infertile and normal rats using isobaric tags for relative and absolute quantitation (iTRAQ) technique. The confidence of proteome data was validated by enzyme-linked immunosorbent assays. RESULTS: 1045 proteins were tested, and 23 of them presented different expression profiling in the infertile and normal rats. The seven proteins were down-regulated, and 16 proteins were up-regulated. Among the seven proteins which were significantly down-regulated by dutasteride in the epididymal luminal fluids, there were three ß-defensins (Defb2, Defb18 and Defb39), which maybe the key proteins involved in epididymal sperm maturation and male fertility. CONCLUSIONS: We report for the first time that dutasteride influences the protein expression profiling in the epididymal luminal fluids of rats, and this result provides some new epididymal targets for male contraception and infertility therapy.

[Inhibitory effect of dutasteride on the expressions of epididymal Claudin1 and ß-catenin in male rats].

OBJECTIVE: To explore the molecular mechanism of dutasteride inhibiting fertility by studying its effects on the expressions of the epididymal epithelial junction proteins Claudin1 and ß-catenin in rats. METHODS: Sixteen 3-month-old SD male rats were equally divided into an experimental and a negative control group to be treated intragastrically with dutasteride at 40 mg/kg per day and the same dose of solvent, respectively, for 14 consecutive days. Then, the sperm motility and morphology of the rats were detected by computer-assisted sperm analysis, the serum levels of testosterone (T) and dihydrotestosterone (DHT) measured by ELISA, changes in the tight junction of epididymal cells observed under the transmission electron microscope, the protein and gene expressions of Claudin1 and ß-catenin determined by RT-PCR and immunohistochemistry, and the conception rate of the mated female rats calculated. RESULTS: Dutasteride significantly suppressed the serum DHT level, sperm motility, and fertility of the rats (P <0.05). Interspaces between epididymal epithelial cell tight junctions were observed, the volume of epididymal fluid obviously increased, and the expressions of Claudin1 and ß-catenin gene and protein remarkably downregulated in the experimental rats (P <0.05). CONCLUSION: Dutasteride can significantly inhibit the fertility of male rats by reducing the serum DHT level, suppressing Claudin1 and ß-catenin expressions, and damaging epididymal epithelial cell junctions.

The purine-carboxylate-containing coordination polymer poly[[µ4-1-(2-carboxylatoethyl)-6-oxo-6,9-dihydro-1H-purin-9-ido]zinc(II)].

The title compound, [Zn(C8H6N4O3)]n or [Zn(L)]n [H2L is 3-(6-oxo-6,9-dihydro-1H-purin-1-yl)propionic acid], crystallized as a nonmerohedral twin. The Zn(II) cation is four-coordinated, ligated by two carboxylate O atoms from two L ligands and two N atoms from another two ligands. Each ligand bridges four Zn(II) centres, extending the structure into a three-dimensional polymer with a 4-connected (6(5),4(1)) topological structure containing two-dimensional homochiral layers constructed from one-dimensional metal-organic helices. Investigation of the thermal stability of the compound shows that the network has very high thermostability and is stable up to 720 K.

Poly[bis[µ2-3-(pyridin-4-yl)benzoato-κ³N:O,O']lead(II)].

In the title compound, [Pb(C12H8NO2)2]n, the Pb atom sits on a crystallographic C2 axis and is six-coordinate, ligated by two chelating carboxylate groups from two 3-(pyridin-4-yl)benzoate (L) ligands and by two N atoms from another two ligands. Each ligand bridges two PbII centres, extending the structure into a corrugated two-dimensional (4,4) net. The ligand L is conformationally chiral, with a torsion angle of 27.9 (12)° between the planes of its two rings. The torsion angle has the same sense throughout the structure, so that the extended two-dimensional polymer is homochiral. Investigation of the thermal stability shows that the network is stable up to 613 K. In the absence of any stereoselective factor in the preparation of the compound, the enantiomeric purity of the crystal studied, based only on the torsional conformation of the ligand, implies that the bulk sample is a racemic conglomerate.

catena-Poly[[[diaquazinc]-bis{µ-N2,N6-bis[(pyridin-3-yl)methyl]pyridine-2,6-dicarboxamide}] dinitrate].

In the title compound, {[Zn(C(19)H(17)N(5)O(2))(2)(H(2)O)(2)](NO(3))(2)}(n), the Zn(II) cation is located at an inversion centre within a slightly distorted octahedron, ligated by four N atoms from four N(2),N(6)-bis[(pyridin-3-yl)methyl]pyridine-2,6-dicarboxamide (L) ligands occupying a plane about the Zn(II) atom with the two water O atoms perpendicular to that. In the complex molecule, the bidentate bridging L ligands display helical R and S conformers, and link the Zn(II) cations into a one-dimensional centrosymmetric double-chain structure containing 32-membered rings. The nitrate anions reside in these rings and are involved in multiple N-H...O hydrogen-bond interactions. On excitation at 390 nm, the title compound displays a strong blue emission centred at 449 nm. Investigation of the thermal stability shows that the network structure is stable up to 420 K.

catena-Poly[[[diaqua[3-(pyridin-4-yl)benzoato-κ2O,O']terbium(III)]-bis[µ-3-(pyridin-4-yl)benzoato-κ2O:O']] monohydrate].

In the title compound, {[Tb(C(12)H(8)NO(2))(3)(H(2)O)(2)]·H(2)O}(n), the Tb(III) cation is in an eight-coordinate environment, ligated by six carboxylate O atoms from five 3-(pyridin-4-yl)benzoate (L) ligands and by two O atoms from water molecules. The cations are bridged by the carboxylate O atoms of the L ligands to form a two-stranded polymeric chain which is assembled into a three-dimensional supramolecular network through regular interchain O-H···N hydrogen bonding. On excitation at 320 nm, the title compound displays a series of emissions, which were assigned to the characteristic electronic transitions of Tb(III).

Poly[[bis{µ2-N2,N6-bis[(pyridin-3-yl)methyl]pyridine-2,6-dicarboxamide}dichloridonickel(II)] N,N-dimethylformamide tetrasolvate].

In the title compound, {[NiCl(2)(C(19)H(17)N(5)O(2))(2)]·4C(3)H(7)NO}(n), the Ni(II) atom is located on an inversion centre and is in a six-coordinated octahedral geometry, formed by four pyridine N atoms from four N(2),N(6)-bis[(pyridin-3-yl)methyl]pyridine-2,6-dicarboxamide (BPDA) ligands occupying the equatorial plane and two chloride anions at the axial sites. The bidentate bridging BPDA ligands link the Ni(II) atoms into a two-dimensional corrugated grid-like flexible layer with a (4,4)-connected topology, which consists of left- and right-handed helical chains sharing the common Ni(II) atoms. Investigation of the thermal stability shows that the network is stable up to 573 K.

catena-Poly[[[diaqua-[3-(pyridin-4-yl)benzoato-κ(2)O,O']gadolinium(III)]-bis-[µ-3-(pyridin-4-yl)benzoato-κ(2)O:O']] monohydrate].

In the title coordination polymer, {[Gd(C(12)H(8)NO(2))(3)(H(2)O)(2)]·H(2)O}(n), the Gd(III) ion is ligated by one bidentate carboxyl-ate group, four monodentate bridging carboxyl-ate O atoms and two water mol-ecules. The resulting GdO(8) polyhedron approximates to a square anti-prism. The bridging ligands link the metal ions into a [100] chain, with each pair of adjacent metal ions being bridged by two ligands. Inter-chain O-H⋯O and O-H⋯N hydrogen bonds help to establish the packing.

catena-Poly[[diaqua-nickel(II)]-bis-(µ-2-{[5-(pyridin-4-yl)-1,3,4-oxadiazol-2-yl]sulfan-yl}acetato)].

In the title compound, [Ni(C(9)H(6)N(3)O(3)S)(2)(H(2)O)(2)](n), the Ni(II) atom, located on an inversion center, is ligated in an octa-hedral geometry by two carboxyl-ate O atoms from two 2-{[5-(pyridin-4-yl)-1,3,4-oxadiazol-2-yl]sulfan-yl}acetate (L) ligands and two O atoms from water mol-ecules in the equatorial plane, and two pyridine N atoms from other two L ligands at the apical sites. Two L ligands bridge pairs of metal atoms in an anti-parallel manner, forming centrosymmetric dinuclear quasi-recta-ngular units which are linked into infinite double-stranded chains parallel to [100]. O-H⋯O hydrogen bonds between the coordinating water mol-ecules and the carboxyl-ate groups of the L ligand as well as interchain S⋯N inter-actions [2.726 (2)-3.363 (2) Å] lead to the formation of a layer structure parallel to (001).

Bis{N(2),N(6)-bis-[(pyridin-3-yl)meth-yl]pyridine-2,6-dicarboxamide-κN}bis-(methanol-κO)bis-(thio-cyanato-κN)cobalt(II).

In the title compound, [Co(NCS)(2)(C(19)H(17)N(5)O(2))(2)(CH(3)OH)(2)], the Co(II) atom lies on an inversion center and is coordinated by two isothio-cyanate N atoms, two O atoms of methanol mol-ecules and two pyridine N atoms in a slightly distorted octa-hedral environment. Inter-molecular O-H⋯O and N-H⋯N hydrogen bonds join the complex mol-ecules into layers parallel to the bc plane.

Tetra-aqua-bis-[3-(pyridin-4-yl)benzoato-κN]manganese(II).

In the title compound, [Mn(C(12)H(8)NO(2))(2)(H(2)O)(4)], the Mn(2+) ion lies on a twofold rotation axis and has a distorted N(2)O(4) octa-hedral coordination geometry formed by four water O atoms in the equatorial plane and two apical pyridyl N atoms. A three-dimensional network is formed in the crystal structure by multiple O-H⋯O hydrogen bonds between the coordin-ating water molecules and the free carboxylate groups.

Tetra-aqua-bis-[3-(pyridin-4-yl)benzoato-κN]cobalt(II).

In the title compound, [Co(C(12)H(8)NO(2))(2)(H(2)O)(4)], the Co atom lies on a twofold rotation axis and has an N(2)O(4) octa-hedral coordination environment formed by four O atoms of water mol-ecules in the equatorial plane and two apical N atoms of pyridine groups. An intricate three-dimensional supra-molecular network is formed by multiple O-H⋯O hydrogen bonds between the coordinated water mol-ecules and the uncoordinated carboxyl-ate groups.

Tetra-aqua-bis-(2-{[5-(pyridin-4-yl)-1,3,4-oxadiazol-2-yl]sulfan-yl}acetato)-cobalt(II) monohydrate.

In the title compound, [Co(C(9)H(6)N(3)O(3)S)(2)(H(2)O)(4)]·H(2)O, the two 2-{[5-(pyridin-4-yl)-1,3,4-oxadiazol-2-yl]sulfan-yl}acetate ligands are monodentate. One coordinates the metal atom via the pyridyl N atom whereas the other coordinates via the carboxyl-ate O atom. The Co(II) atom adopts a slightly distorted octa-hedral coordination geometry with four O atoms of the coordinated water mol-ecules located in the equatorial plane and the N and O atoms of the two POA ligands in axial positions. In the crystal, the components are connected through O-H⋯O and O-H⋯N hydrogen bonds into a three-dimensional framework.

Tetra-aqua-bis-(2-{[5-(pyridin-4-yl)-1,3,4-oxadiazol-2-yl]sulfan-yl}acetato)-iron(II).

In the title compound, [Fe(C(9)H(6)N(3)O(3)S)(2)(H(2)O)(4)] or [Fe(POA)(2)(H(2)O)(4)], the Fe(II) atom is located on an inversion center and is ligated by four O atoms of coordinated water mol-ecules in the equatorial plane while two POA ligands acting as monodentate ligands occupy the axial positions through their pyridyl N atoms, completing a slightly distorted octa-hedral coordination geometry. A three-dimensional supra-molecular network is formed by multiple O-H⋯O hydrogen-bonding inter-actions between the coordinated water donors and the uncoordinated carboxyl acceptors.

[Enhanced decomposition of 1, 4-benzoquinone ring by DSA electrode under ultraviolet irradiation].

Commercial DSA electrode was employed for photoelectrochemical degradation of a model pollutant 1,4-benzoquinone. The effect of electrolyte, electrical field intensity and solution pH conditions were investigated. The results proved that the TOC removal by photoelectrochemical oxidation was 1.25 times that of photocatalytic oxidation and electrocatalytic oxidation alone, indicating a synergetic effect. Addition of electrolyte and application of external electrical field have effectively enhanced the photoelectrochemical oxidation efficiency within a certain range of concentration and intensity. Acidic and neutral pH conditions are favorable for ring opening of 1,4-Benzoquinone.The photoelectrochemical oxidation mechanism is also discussed. It suggests that the efficiency of electrochemically assisted photoelectrochemical oxidation process can be manipulated, accompanied by the generation of hydroxyl radicals, which will be more adaptable for water and wastewater treatment.

Lutetium(III)-dependent self-assembly study of ciliate Euplotes octocarinatus centrin.

Ciliate Euplotes octocarinatus centrin (EoCen) is a member of the EF-hand superfamily of calcium-binding proteins, which often associated with the centrosomes and basal bodies. To explore the possible structural role of EoCen, we initiated a physicochemical study of the self-assembly properties of the purified protein in vitro. The native PAGE results indicate that only the integral protein shows multimers in the presence of Lu(3+). The dependence of Lu(3+) induced self-assembly of EoCen on various chemical and physical factors, including temperature, protein concentration, ionic strength and pH, was characterized using resonance light scattering (RLS). Control experiments with different metal ions suggest that Ca(2+) and Lu(3+) bindings to the N-terminal domain of EoCen are all positive to the self-assembly of the protein, and Lu(3+) exhibits the stronger effect, however, Mg(2+) alone cannot take the same effect. The experiments of 2-ptoluidinylnaphthalene-6-sulfonate (TNS) binding and ionic strength demonstrate that the lutetium(III)-dependent self-assembly is closely related to the exposure of hydrophobic cavity. Control experiment on pH value with EoCen and the fragments of it, N-terminal domain of EoCen (N-EoCen), indicates that the electrostatic effect is of small tendency to be served as the main driving force in the self-assembly of EoCen. The specific oligomerization form of the protein was exhibited by cross-linking experiment.

Investigation on the binding of TNS to centrin, an EF-hand protein.

The interaction between 2-p-toluidinylnaphthalene-6-sulfonate (TNS) and ciliate Euplotes Octocarinatus centrin (Cen) has been studied by fluorescence spectroscopy. The binding constants of TNS with Cen were measured at different temperature in the 0.01M Hepes, pH 7.4. The binding process is exothermic and involves a positive entropy change. The negative value of enthalpy predominately contributes to the negative free energy of binding between TNS and Cen. The salt (KCl) increases the association constant of TNS and Cen. These results and resonance light scattering experiment suggest that the binding force between TNS and Cen is hydrophobic. The distance (r) between TNS and tryptophan of mutant G115W, which sheds more insight into the binding of TNS to Cen, was determined as 4.85nm based on Förster non-radiative energy transfer theory.

Process conditions for preparing methanol from cornstalk gas.

The low-heat-value cornstalk gas produced in the down-flow fixed bed gasifier was tentatively used for methanol synthesis. The cornstalk gas was purified and the technical procedures such as deoxygenation, desulfurization, catalytic cracking of tar, purification and hydrogenation were studied. The catalytic experiments of methanol synthesis with cornstalk syngas were carried out in a tubular-flow integral and isothermal reactor. The effect of reaction temperature, pressure, catalyst types, catalyst particle size, syngas flow at entering end and composition of syngas was investigated. The optimum process conditions and yield of methanol from cornstalk syngas were obtained. The experimental results indicated that the proper catalyst for the synthetic reaction was C301 and the optimum catalyst size was 0.833 mm x 0.351 mm. The optimum operating temperature and pressure were found to be 235 degrees C and 5 MPa, respectively. The suitable syngas flow 0.9-1.10 mol/h at entering end was selected and the best composition of syngas were CO 10.49%, CO2 8.8%, N2 37.32%, C(n)H(m) 0.95% and H2 40.49%. The best methanol yield was 0.418 g/g cornstalk. This study provided the technical support for the industrial test of methanol production from biomass (cornstalk) gas.