OBJECTIVE To investigate the effects and mechanism of atractylodin on inflammatory injury of periodontal tissue and alveolar bone loss in periodontitis rats. METHODS A total of 144 SD rats were divided into control group （intragastric and intraperitoneal injection of normal saline）， model group （intragastric and intraperitoneal injection of normal saline）， atractylodin low-dose， medium-dose and high-dose groups （intraperitoneal injection of 6.665， 13.33， and 26.66 mg/kg atractylodin）， metronidazole group （positive control group， intragastric injection of 0.05 g/kg metronidazole， intraperitoneal injection of normal saline）， AMD3100 [stromal cell-derived factor-1 （SDF-1）/CXC chemokine receptor-4 （CXCR4） pathway inhibitor] group （intragastric injection of 1 mg/kg AMD3100， intraperitoneal injection of normal saline）， atractylodin high-dose+AMD 3100 group （intraperitoneal injection of 26.66 mg/kg atractylodin， intragastric injection of 1 mg/kg AMD3100）， with 18 rats in each group. Except for the control group， all other groups of rats were inoculated with Porphyromonas gingivalis to construct a periodontitis model. After successful modeling， they were given relevant medicine or normal saline， once a day， for 4 consecutive weeks. The gingival index of rats was detected； the levels of interleukin-6 （IL-6） and tumor necrosis factor α （TNF-α） in rat serum were also determined； alveolar bone resorption， periodontal histopathologic changes and the number of osteoclasts were detected by methylene blue staining， HE staining and TRAP staining， respectively. The expressions of osteoprotegerin （OPG）， receptor activator of NF-κB ligand （RANKL）， SDF-1 and CXCR4 proteins were determined. RESULTS Compared with the control group， serious pathological injury of periodontal tissue was found in the model group， the gingival index， the levels of IL-6 and TNF- α， alveolar bone absorption value， the number of osteoclasts， and the expression of RANKL protein were all increased significantly （P＜0.05）， while the expressions of OPG， SDF-1 and CXCR4 proteins were decreased significantly （P＜0.05）. Compared with the model group， pathological injury of periodontal tissue in rats was reduced； the gingival index， the levels of IL-6 and TNF-α， alveolar bone resorption value， osteoclast number and RANKL protein expression were decreased significantly， while protein expressions of OPG， SDF-1 and CXCR4 were increased significantly in atractylodin low-dose， medium-dose and high-dose groups and metronidazole group （P＜0.05）. The change trend of corresponding indexes in the AMD3100 group was opposite to the above （P＜0.05）. AMD3100 attenuated the inhibitory effect of high-dose atractylodin on inflammatory response and alveolar bone loss in rats with periodontitis （P＜0.05）. CONCLUSIONS Atractylodin may improve the inflammatory response and alveolar bone loss in periodontitis rats by activating the SDF-1/CXCR4 signaling pathway.

Converting two poorly water-soluble crystalline drugs to co-amorphous drug systems by ball milling, quench-cooling, or cryo-milling method can improve stability of the drug, enhance dissolution rates, and reduce adverse reactions of the single drug. Co-amorphous system has been used to solve problems of co-administration of medicines. Formation and intermolecular interactions of co-amorphous drug systems may be verified by differential scanning calorimetry (DSC), X-ray powder diffraction (XRPD), Raman spectroscopy (RS) and Fourier transform infrared spectroscopy (FT-IR). Stability of co-amorphous drug systems is influenced by their glass transition temperature (Tg) and intermolecular interactions. The theoretical Tg values and the interaction parameter x are calculated by Gordon-Taylor equation and the Flory-Huggins equation, respectively. Thus, co-amorphous drug systems are analyzed theoretically at molecular level. Co-amorphous drug systems provide a new sight for the co-administration of medicines.
Calorimetry, Differential Scanning ; Chemistry, Pharmaceutical ; methods ; Cimetidine ; chemistry ; Drug Combinations ; Drug Compounding ; Drug Stability ; Glipizide ; chemistry ; Indomethacin ; chemistry ; Naproxen ; chemistry ; Ranitidine ; chemistry ; Simvastatin ; chemistry ; Solubility ; Spectroscopy, Fourier Transform Infrared ; Spectrum Analysis, Raman ; Technology, Pharmaceutical ; methods ; Temperature ; X-Ray Diffraction

Objective To prepare biphasic calcium phosphate/polyvinyl alcohol scaffolds by 3D printing at room temperature and explore the effect of 3D scaffolds on in vitro osteogenic differentiation of the bone marrow mesenchymal stem cells (BMSCs).Methods After biphasic calcium phosphate and polyvinyl alcohol solutions were mixed,the biphasic calcium phosphate/polyvinyl alcohol composite scaffolds were prepared by room temperature 3D printing combined with freeze drying technique.Non-printing scaffolds were prepared by injection molding.The surface microstructure,porosity,elastic modulus and hydrophilicity of the 2 sorts of scaffolds were measured.The cytological experiments were carried out in 3 groups (n =3):printed scaffold group,non-printed scaffold group and blank control group (no scaffold).After the BMSCs were seeded onto the scaffolds for 7 and 14 days,the 3 groups were compared in terms of cellular proliferation,alkaline phosphatase activity and expression levels of osteogenesis-related genes.Results 3D composite scaffolds with controllable pore size and porosity were prepared successfully,with an average porosity of 59.6％ ± 3.6％ and an average elastic modulus of 429.3 ± 54.3 kPa.After culture for 7 and 14 days,the cellular absorbance values in the printed scaffold group (0.987 ± 0.047 and 1.497 ± 0.076) were significantly higher than those in the non-printed scaffold group (0.767 ±0.063 and 1.181 ±0.098) (P ＜ 0.05) which were in turn significantly higher than those in the blank control group (0.532 ±0.046 and 0.895 ± 0.062) (P ＜ 0.05).After culture for 7 and 14 days,the ALP activity and expression levels of osteogenesis-related genes in the printed and non-printed scaffold groups showed no significant between-group differences (P ＞ 0.05),but were significantly higher than those in the blank control group (P ＜ 0.05).Conclusions Tissue-engineered composite biphasic calcium phosphate/polyvinyl alcohol scaffolds with controllable pore size and good connectivity can be prepared by freeze-drying and room temperature 3D printing techniques.Co-culture of the scaffolds and BMSCs in vitro promotes adhesion,proliferation and osteogenic differentiation of the cells.

OBJECTIVETo study the chemical constituents from Aeschynanthus longicaullis.

METHODThe column chromatographic techniques were applied to isolate the constituents. The spectroscopie methods were used of EI-MS and NMR to identify the structures of the separated compounds.

RESULTSeven compounds were isolated from the ethyl acetate extract of A. longicaullis, whose structures were elucidated as cryptomeridiol (1), 4 (15) -eudesmene-1beta, 6alpha-diol (2), 2,5-bornanediol (3), isovanillic acid (4), vanillic acid (5), stigmast-5 (6) , 22 (23)-diene-3beta-ol (6) and beta-sitosterol (7).

CONCLUSIONCompounds 1-5 were isolated for the first time from Gesneriaceae and 6-7 were isolated for the first time from this plant.

OBJECTIVETo study the chemical constituents from Corallodiscus kingianus.

METHODThe column chromatographic techniques were applied to isolate constituents. A combination of ESI-MS and NMR spectroscopy was used to identify structures.

RESULTSix compounds were isolated from the acetone extract of this plant, and the structures of them have been identified as castanopsol (1) , 3beta-hydroxy-9 (11), 12-oleanadien-28-oic acid (2), 2,3,7-trihydroxy-6-oxo-1,3,5 (10), 7-tetraene-24-nor-frie-delane-29-oic acid methylester (3), 3-epicyclomusalenol (4), palmitinic acid (5) and stearic acid (6).

CONCLUSIONCompounds 1-6 were isolated for the first time from Gesneriaceae.

Objective To assess the clinical threatment results of pilon fractures managed with arthroscopyassisted minimally invasive percutaneous plate osteosynthesis (MIPPO).Methods 33 patients with pilon fractures were classified into 3 groups according to the Ruedi-Allgower classification:type Ⅰ in 26 cases,type Ⅱ in 5 cases,type Ⅲ in 2 cases,including 29 males and 4 females,aged 22 to 51 years,mean 31.5 years of age.All patients were treated with arthroscopy-assisted MIPPO with the postoperative follow-up time of 12 to 84 months.Results The clinical surgery efficacy according to Mazur's criterion was evaluated as excellent in 22 cases,good in 8 cases,fair in 3 cases.The excellent and good rate was 90.9％.Conclusion Arthroscopy-assisted MIPPO surgical treatment is an effective method for Pilon fractures with the advantages of good healing,minimal trauma and less complications,it is worthy of clinical application.

Objective To prepare a bionic artificial bone scaffold using a room temperature three dimensional (3D) printing technique and evaluate its biocompatibility and bioactivity in vitro.Methods A room temperature 3D printing technique was applied to fabricate 3D bionic artificial bone scaffolds using collagen/hydroxyapatite.The physico-chemical structure,porosity and mechanical strength of the scaffolds were assessed.The extract liquid of scaffolds was cocultured with bone mesenchymal stem cells (BMSCs) to evaluate the toxicity of scaffolds.There were 3 experimental groups:blank control with no scaffolds,printed scaffolds group and non-printed scaffolds group.The condition of BMSCs on the scaffolds was observed via scanning electron microscopy(SEM) and immunostaining.3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay and SEM were applied to monitor the proliferation of BMSCs on the scaffolds.At last,alkaline phosphatase (ALP) activity and mRNA expression levels of osteogenesis-related genes were detected to assess the osteoinductive property of the scaffolds.Results The 3D printed scaffolds fabricated in the present study were characterized by highly interconnected pores which were controllable and even in size.The cross section of the scaffolds presented an irregular honeycomb-like microstructure.The porosity of printed 3D scaffolds (71.14％ ± 2.24％) was significantly higher than that of non-printed scaffolds (59.04％ ±2.98％) (P ＜ 0.05).The physico-chemical structures of the materials were preserved after printing without additional cytotoxicity.The MTT results at 7 and 14 days revealed that the printed scaffolds had a significantly more cell numbers than the non-printed scaffolds(P ＜ 0.05).SEM showed that the BMSCs adhered well onto the printed scaffolds and proliferated and migrated through the pores.Compared with the blank control,the printed scaffolds showed obviously better osteogenic outcomes.Conclusion The 3D bionic artificial bone scaffolds of collagen/hydroxyapatite manufactured by a room temperature 3D printing technique can provide a good extracellular matrix for BMSCs to proliferate and differentiate.

To study the heating treatment of Angelica dahurica under different temperature and time conditions on the stability of coumarins and tyrosinase activity. HPLC method was used to determine the contents of imperatorin and isoimperatorin, and tyrosinase activity was assayed by measuring the oxidation rate of L-DOPA in vitro. After heated, the contents of imperatorin increased. Expect for being heated at 90 ℃ for 2 h, the content of isoimperatorin was higher than crude one. Before and after being heated, A. dahurica showed an activating effect on tyrosinase. In the same temperature and time conditions, the activation rate increased with the rise of concentration of tyrosinase extracts. Heating process for A. dahurica could change the contents of imperatorin and isoimperatorin, mainly increasing their concentrations.

Hypercoagulable state and thrombosis are major lethal causes of ulcerate colitis (UC). The aim of the present study is to explore the change and role of protein C (PC) system in UC thrombosis. 4% dextran sulfate sodium (DSS) was used to induce the UC model, and the body weight, the length of colon, and the weight of spleen were measured after intake of DSS as drinking water for 1 week. The macroscore and microscore were examined. The quantity of macrophage in colon smooth muscle was observed by immunofluorescence, and TNF-α and IL-6 levels in plasma were evaluated by ELISA. Intravital microscopy was applied to observe colonic mucosal microvascular circulation, activities of PC and protein S (PS) were determined by immunoturbidimetry, endothelial cell protein C receptor (EPCR) and thrombomodulin (TM) expressions were detected by immunohistochemistry. In vitro, TNF-α and IL-6 levels were tested in supernatant of macrophage separated from colonic tissue. After stimulation of mouse colonic mucosa microvascular endothelial cells by TNF-α and IL-6 respectively, the activities of PC, PS, activated protein C (APC) were evaluated, and the expressions of EPCR and TM were detected by Western blotting. The results revealed that compared with control, the DSS mouse showed weight loss (P < 0.05), a shortened colon (P < 0.05), and swelled spleen (P < 0.05), accompanied by higher histological score (P < 0.05), as well as infiltration of macrophages, elevated TNF-α and IL-6 levels in plasma (P < 0.01). The intravital microscopy results revealed that compared with control, DSS mice showed significantly enhanced adhesion of leukocytes and colonic mucosal microvascular endothelial cells (P < 0.01), meanwhile, decreased activity of PC and PS in plasma (P < 0.01 or P < 0.05), and down-regulated expression of EPCR (P < 0.01). The degree of inflammation was negatively correlated with the PC activity. In vitro, TNF-α and IL-6 levels were increased in the supernatant of macrophages from DSS mice colonic tissue (P < 0.05), and after incubation of TNF-α or IL-6 with colonic mucosal microvascular endothelial cells, the APC activity was decreased (P < 0.05 or P < 0.01), and expression of EPCR was down regulated (P < 0.05). These results suggest that PC system is inhibited in UC mouse. Presumably, the mechanism may be due to the secretion of cytokines from macrophages and subsequential influence on the function of endothelia cells. Furthermore, enhancement of PC system activity may serve as a new strategy for the treatment of UC.
Animals ; Blood Coagulation Factors ; metabolism ; Colitis, Ulcerative ; chemically induced ; physiopathology ; Dextran Sulfate ; Immunohistochemistry ; Inflammation ; Interleukin-6 ; blood ; Intestinal Mucosa ; pathology ; Macrophages ; cytology ; Mice ; Protein C ; metabolism ; Receptors, Cell Surface ; metabolism ; Spleen ; pathology ; Tumor Necrosis Factor-alpha ; blood

The study is aimed to explore the molecular mechanism of the treatment of apocynin in dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) mice. 5% DSS was used to mimic the UC model, and 2% apocynin was applied to treat the UC mice. HE staining was used for histopathological evaluation. Chemiluminescence technique was used to measure reactive oxygen species (ROS) production, and the rate of consumption of NADPH inhibited by DPI was detected to determine the NADPH oxidases (NOXs) activity. Western blot was applied to identify the level of p38MAPK phosphorylation, Griess reaction assay to analyze NO production, immunoenzymatic method to determine prostaglandin E2 (PGE2) production, real time RT-PCR and Western blot to identify the expression of iNOS and COX2, and enzyme linked immunosorbent assay to detect inflammatory cytokines TNF-α, IL-6, IFN-γ, IL-1β. Rat neutrophils were separated, and then ROS production, NOXs activity, NO and PGE2 production, NOX1 and p-p38MAPK expression were detected. Compared with the UC group, apocynin decreased ROS over-production and NOXs activity (P < 0.01), reduced p38MAPK phosphorylation, inhibited NO, PGE2 and cytokines production (P < 0.01). Apocynin also decreased NOXs activity and ROS over-production (P < 0.01), inhibited p38MAPK phosphorylation and NOX1 expression, and reduced NO and PGE2 production (P < 0.01) in separated neutrophils from UC mice. Therefore, apocynin could relieve inflammation in DSS-induced UC mice through inhibiting NOXs-ROS-p38MAPK signal pathway, and neutrophils play an important role.
Acetophenones ; pharmacology ; Animals ; Colitis, Ulcerative ; chemically induced ; drug therapy ; Cytokines ; metabolism ; Dextran Sulfate ; Inflammation ; drug therapy ; MAP Kinase Signaling System ; Mice ; NADH, NADPH Oxidoreductases ; metabolism ; Neutrophils ; metabolism ; Rats ; Reactive Oxygen Species ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism