Objective To investigate the effects of warming-needle acupuncture on the levels of matrix metalloproteinase-1 (MMP-1) and interleukin-1β (IL-1β) in the articular cartilage in knee osteoarthritis induced by immobilization in rabbits.MethodsA total of 30 adult male Japanese white rabbits were randomly assigned into three groups by random number table method: a normal group, a model group and a treatment group, with 10 rabbits in each group. Knee osteoarthritis was induced by 6 weeks of immobilization. The rabbits in the treatment group received warming-needle acupuncture after modeling at points of “Neixiyan” (EX-LE4), “Dubi” (ST35), “Zusanli” (ST36) and “Yanglingquan” (GB34) for 4 weeks. The level of MMP-1 and IL-1β in the articular cartilage of the knee were determined with radioimmunological assay.ResultsThere were significant difference in the levels of MMP-1(0.16 ± 0.02 ng/mg, 0.37 ± 0.02 ng/mg, and 0.28 ± 0.03 ng/mg;F=258.251) and IL-1β (0.21 ± 0.01 pg/mg, 0.34 ± 0.02pg/mg, and 0.31 ± 0.04 pg/mg;F=127.112) among the normal group, model group and treatment group. The levels of MMP-1 and IL-1β both in the model group and treatment group were significant higher than those in the normal group (allP<0.01), while the levels of MMP-1 and IL-1β in the treatment group were significant lower than those in the model group (allP<0.01).ConclusionWarming-needle acupuncture can effectively reduced the levels of MMP-1 and IL-1β in the articular cartilage in knee osteoarthritis induced by immobilization in rabbits.

UNLABELLEDOBJECTIVE To investigate the effect of Nepsilon-(carboxymethyl) lysine albumin (CMLs), a primary advanced glycation end products (AGEPs) isoform in diabetic body, on the function and angiogenesis of adipose tissue-derived stem cells (ADSCs) and the protective effect of Danhong Injection (DH). METHODS Human ADSCs were cultured and separated from human subcutaneous fatty tissue using enzymatic digestion and centrifugation. The morphology was observed using optical microscope and differentiation capacities assessed. Cells were exposed to 5 different interventions respectively for 24 h, i.e., PBS, 60 1 microg/mL BSA, 60 microg/mL CML-BSA, 100 microL/mL DH, and 60 micro./mL CML-BSA +100 microL/mL DH. Their effect on the proliferation, migration, apoptosis, and secretion were observed using WST-1 assay, Transwell assay, Annexin V-FITC/PI flow meter test reagent kit, human VEGF reagent kit, ELISA reagent kit, respectively. The effect on ADSCs angiogenesis was observed by in vitro angiogenesis test.

RESULTSCompared with the BSA group, the capacities of proliferation and migration could be significantly inhibited by CML-BSA, the apoptosis promoted, the secretion of VEGF reduced, and the angiogenesis of ADSCs weakened (P < 0.05). Compared with the blank control group, 100 microL/mL DH could significantly promote the proliferation and migration capacities of ADSCs, inhibit apoptosis of ADSCs, increase the secretion of VEGF, and improve the angiogenesis of ADSCs (P < 0.05). Compared with the CML-BSA group, the inhibition of CML-BSA on the proliferation and migration capacities of ADSCs could be significantly reversed, the promotion of CML-BSA on the apoptosis of ADSCs improved, the secretion of VEGF increased, and the angiogenesis of ADSCs elevated (P < 0.05).

CONCLUSIONclusion CMLs could significantly inhibit the proliferation and migration capacities of ADSCs, promote their apoptosis, and inhibit their angiogeneses, which could be improved by DH.

Adipose Tissue ; cytology ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Glycation End Products, Advanced ; pharmacology ; Humans ; Neovascularization, Pathologic ; drug therapy ; Stem Cells ; cytology ; drug effects

Objective To investigate the roles of dCK Ser-74 in radiation-induced cell death in breast cancer cells.Methods Different phenotypes of dCK plasmids were transfected into MCF-7 cells by liposome transfection,including dCK-Vector,dCK-WT (wild type),dCK-S74A (non-phosphorylation) and dCK-S74E (hyper-phosphorylation).All these cells were irradiated by 0,2,4,6,8 Gy X-rays,respectively.The transcriptional and translational level of dCK were detected with real time-PCR and Western blot,respectively.Radiosensitivity was analyzed using cell counting kit (CCK-8) and colony formation assays.Monodansylcadaverine staining (MDC) and flow cytometry were used to detect autophagy and apoptosis,respectively.Results Four phenotypes of dCK cell models were established successfully.After irradiation,the cell viabilities of MCF-7 and dCK-Vector decreased significantly as compared with mock group (t =14.469 and 9.357,P ＜ 0.05),the cell viabilities of dCK-WT,dCK-S74A and dCK-S74E showed no changes (P ＞ 0.05).The total mortalities of dCK-WT and dCK-S74E decreased significantly as compared with dCK-Vector (x2 =3.857-3.971,P ＜ 0.05),but no changes in dCK-S74A cells (P ＞0.05).The apoptosis rates in dCK-S74A,dCK-Vector and control group were up-regulated after irradiation (t =-4.531,-3.688 and-7.076,P ＜ 0.05),and the irradiation-induced apoptosis was reversed in dCK-WT and dCK-S74E (66％ and 68％ of the increase level in dCK-Vector group).The autophagy in dCK-WT and dCK-S74E increased by 22％ and 26％ (t =-9.051 and-8.411,P ＜0.01),but no changes were observed in dCK-S74A,dCK-Vector and control groups (P ＞ 0.05).Conclusions The dCK-WT and dCK-S74E could reverse the irradiation-induced apoptosis,increase the autophagy occurence,and decrease the total mortality,indicating that the phosphorylation of dCK at Ser-74 sites is related to the radiosensitivity of MCF-7 cells.

Although current synthetic anti-gout drugs have significant therapeutic effects in reducing serum uric acid levels, they have serious side effects such as allergic reactions and liver and kidney damage. Natural products with a wide range of uric acid-lowering and high safety have played a critical role in anti-gout drug discovery and development. This paper reviews the natural products with uric acid-lowering or anti-gout pharmacological effects and the investigation on their mechanisms of action, to provide information for drug discovery and development.

Objective To analyze the worldwide advances on bacterial quantitative proteomics over the past fifteen years with bibliometric approach.Methods Literature retrieval was conducted throughout the databases of Pubmed,Embase and Science citation index (SCI),using bacterium and quantitative proteomics as the key words.The deadline is July 2013.We sorted and analyzed these articles with Endnote X6 from the aspects of published year,the first author,name of journal,published institution,cited frequency and publication type.Results 932 English articles were included in our research after deleting the duplicates.The first article on bacterial quantitative proteomics was reported in 1999.The maximal publications were 163 related articles in 2012.Up till July 2013,authors from more than 23 countries and regions have published articles in this field.China ranks the fourth.The main publication type is original articles.The most frequently cited article is entitled with Absolute quantification of proteins by LCMSE:a virtue of parallel MS acquisition by Silva JC,Gorenstein MV,Li GZ,et al in Mol Cell Proteomics 2006.The most productive author is Smith RD from Biological Sciences Division,Pac.Northwest National Laboratory.The top journal publishing bacterial quantitative proteomics is Proteomics.Conclusion More and more researchers pay attention to quantitative proteomics which will be widely used in bacteriology.

Objective To investigate the combination effect using iron chelators deferasirox and cisplatin on A549 cell proliferation and apoptosis and to provide evidences to explore an effective way to treat lung cancer. Methods Lung adeno?carcinoma cells were cultured by conventional way, with administration of different concentrations of deferasirox and cisplat?in. Cell growth inhibition was observed under an inverted microscope. Proliferation inhibition was evaluated by MTT assay. Morphological changes of cell apoptosis was detected using DAPI,AO/EB straning and flow cytometry. Results After a cer?tain time of incubation with different concentrations of the combined drugs,the cell number reduce significantly, which was counted under invert microscope. Cells were dispersed with each other and adherent cells appear shrunken and poor in re?fractivity. MTT assay showed that inhibition of cell proliferation was in a concentration-time-dependent manner. Chromatin condensation, nuclear condensation and other typical apoptotic morphology were detected after DAPI and AO/EB straning. Flow cytometry showed that apoptosis increased with rising drug concentration. So combination therapy was significantly pro-apoptotic. Conclusion Deferasirox has the ability to inhibit proliferation of A549 cells and can promote tumor cell apoptosis and enhances cancer cell tolerance when combined with cisplatin. It can also reduce the amount and toxicity of cis?platin. It provides a basis for finding an effective way to treat lung cancer.

Objective To determine whether the susceptibility to ischemia-reperfusion injury in aged heart is higher than that in adult heart and,if so,to clarify the mechanisms underlying this change.Methods Wister rats(5-or 20-month-old)were randomly divided into 4 groups(6 animals in each group).The rats were subjected to 30 minutes of myocardial ischemia via ligating the left anterior descending coronary artery,followed by 3 hours of reperfusion(Young-MI/R group and Old-MI/R group);A silk suture around the left anterior descending coronary artery was not ligated in young and old rats(Young-sham group and Old-sham group).Myocardial apoptosis was detected by terminal deoxynueleotidyl transferase biotin-d UTP nick end labeling(TUNEL)staining and caspase-3 activity was detected by using a caspase-3 colorimetrie assay.Nitrotyrosine content,a footprint of in vivo ONOO~-formation,and total NO content were determined by ELISA and chemiluminescence method respectively.Results A significantly exacerbated cardiac reperfusion injury was found in Old-MI/R group as evidenced by increased TUNEL positive myocytes[(19.0?2.1)% vs.(14.6?1.7)%],and increased myocardial caspase-3 activity[(436?35)?mool/mg vs.(340?32)?mol/mg] compared with Young-MI/R group(P＜0.05).Aged hearts had a markedly increased basal NOx level compared with young adult hearts.Marked higher myocardial nitrotyrosine content was found in OId-MI/R group[(7.25?0.18)nmol/g]than that in Young-MI/R group[(4.68?0.15)nmol/g] (P＜0.05).Conclusions In aged hearts,high levels of NO might form highly toxie derivant, ONOO~-,and its subsequent nitrified protein.This may attribute to the increased susceptibility of the aged heart to isehemic-reperfusion injury.

Fatigue is a common complication of type 2 diabetes mellitus (T2DM). We examined the relationship between T2DM fatigue and the skeletal muscle 5-hydroxytryptamine (5-HT) system. In animal experiments, a T2DM model was established in mice by feeding a high-fat diet with intraperitoneal injection of streptozotocin. The mice were treated with the 5-HT_2A receptor antagonist sarpogrelate hydrochloride (SH) and the 5-HT synthesis inhibitor carbidopa (CDP) (separately and in combination). In cell culture experiments, C2C12 cells were stimulated with D-glucose, palmitic acid or 5-HT. 5-HT_2AR, 5-HT synthesis and 5-HT degradation were inhibited by SH, CDP, or monoamine oxidase A (MAO-A) inhibitor. The animal experiments were in accordance with the regulations of the Animal Ethics Committee of China Pharmaceutical University. The results showed that 5-HT_2AR, 5-HT synthase and MAO-A were expressed in mouse skeletal muscle and C2C12 cells. The expression of these proteins was significantly up-regulated in T2DM mice or when C2C12 cells were exposed to palmitic acid and D-glucose; palmitic acid was a stronger stimulant of their expression than D-glucose. Rotating rod experiments and biochemical index tests have shown that T2DM fatigue is associated with an increase in skeletal muscle 5-HT_2AR, 5-HT synthesis and 5-HT degradation. 5-HT_2AR mediates the expression of MAO-A and the synthesis of 5-HT, which indirectly regulates the degradation of 5-HT. MAO-A regulates cell inflammation, mitochondrial ROS production and membrane potential depolarization by mediating 5-HT degradation. MAO-A also inhibits the expression of peroxisome proliferator-activated receptor γ coactivator-1 (PGC-1), carnitine palmitoyltransferase-1 (CPT1) and ATP synthase-6 (ATP6), thus inhibiting mitochondrial functions such as fatty acid β oxidation and ATP synthesis. SH and CDP can effectively treat T2DM fatigue, and can also reduce blood glucose and blood lipid, and the combination of SH and CDP has a clear synergistic effect.

Antioxidants ; pharmacology ; Ascorbic Acid ; pharmacology ; Carbon Disulfide ; adverse effects ; antagonists & inhibitors ; Cells, Cultured ; Endothelium, Vascular ; drug effects ; Humans ; Umbilical Veins ; cytology ; drug effects ; metabolism

OBJECTIVETo study the chemical constituents possessing cytotoxicity activity from Elaeagnus pungens.

METHODThe constituents were separated through repeated chromatographic methods and their structures were elucidated by spectral analysis.

RESULTFive compounds were isolated from the ethyl acetate ether extract of leaves of E. pungens. Their structures were elucidated as 4-hydroxybenzoic acid (1), 3, 3'-dimethoxyquercetin (2), caffeic acid methyl ester (3), methyl 3, 4-dihydroxybenzoate (4), spingic acid (5), 4-methoxylbenzoic acid (6), 3-methylkaempferol (7), kaempferol-3-O-beta-D-glucoside (8), dausosterol (9).

CONCLUSIONCompounds 1-8 were isolated from this plant for the first time.

Caffeic Acids ; chemistry ; isolation & purification ; pharmacology ; Cell Proliferation ; drug effects ; Elaeagnaceae ; chemistry ; HeLa Cells ; cytology ; Humans ; Kaempferols ; chemistry ; isolation & purification ; pharmacology ; Parabens ; chemistry ; isolation & purification ; pharmacology ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Quercetin ; analogs & derivatives ; chemistry ; isolation & purification ; pharmacology