In temporomandibular disorders (TMD), pain takes place when neuropeptides stimulate synovial tissue to produce several cytokines such as interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α, which activate neurons and glia of synovial membrane at the bilaminar regions of temporomandibular joint (TMJ). It has been reported that, after neurogenic differentiation, the synovial mesenchymal stem cells (SMSCs), deriving from TMJ, possess the same cytological features as the neuronal cells. This study examined the ability of substance P (SP) and calcitonin gene-related peptide (CGRP) to stimulate SMSCs and neurogenic SMSCs secreting inflammatory cytokines during TMD, evaluated the mutual effects of inflammatory cytokines and neuropeptides and tested the analgesic effect of hyaluronic acid (HA). The levels of IL-1β, IL-6 and TNF-α in SMSCs and neurogenic SMSCs in the presence of neuropeptides were measured by ELISA. SP and CGRP produced by SMSCs and neurogenic SMSCs were determined by RT-PCR and Western blotting. The results showed that the expression of SP and CGRP was significantly enhanced in the neurogenic SMSCs in response to IL-1β, IL-6 and TNF-α, and the effect was remarkably inhibited by HA. IL-1β, IL-6 and TNF-α, in return, could be enhanced in the neurogenic SMSCs upon stimulation by SP and CGRP. Neuropeptides and inflammatory cytokines might work mutually on the TMD pain. The HA-mediated analgesic effect may be implicated in the inhibition of SP and CGRP expression in neurogenic SMSCs.

A method of LC-QTOF/MS combining with chemical synthesis has been used to determine the structures of three novel bile acids from bear bile powder. Reference substances of tauroursodeoxycholic acid and taurochenodeoxycholic acid were oxidized by pyridinium chlorochromate. The products were analyzed by LC-QTOF/MS. Total 4 products including 3 isomers were predicted and identified according to the PCC oxidation theory and LC-QTOF/MS results. Bear bile powder samples were dissolved by methanol and analyzed by LC-QTOF/MS. Three unknown peaks were found and identified as 2-[[(3beta, 5beta)-3-hydroxy-7, 24-dioxocholan-24-yl]amino]-ethanesulfonic acid, 2-[[(5beta)-3, 7, 24-trioxocholan-24-yl]amino]-ethanesulfonic acid and 2-[[(5beta, 7beta)-7-hydroxy-3, 24-dioxocholan-24-yl]amino]-ethanesulfonic acid, separately, by matching their results with that of oxidation products above.
Animals ; Bile ; chemistry ; Bile Acids and Salts ; analysis ; chemistry ; Chromatography, Liquid ; methods ; Isomerism ; Molecular Structure ; Oxidation-Reduction ; Powders ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods ; Taurochenodeoxycholic Acid ; chemistry ; Ursidae

Objective To investigate the relationship between insulin resistance and erythrocyte in- sulin receptors in patients with gout associated with macroalbruminuria(MAU).Methods FBG,PBG,FINS, P2hINS,CH,TG,HDL,LDL-c,UA and erythrocyte insulin receptors were determined in 44 patients with MAU,62 patients with normal MAU(NMAU).Results In MAU,the levels of FINS,TG,LDL-c and HOMA-IR were(16?4)mU/L,(2.5?0.6)retool/L,(3.2?0.5)mmol/L and 3.6~1.2 respectively.While they were(13?3) mU/L,(2.3?0.8)mmol/L,(3.0?0.5)mmol/L and 3.0~0.4 in NMAU group.The levels of FINS,TG,LDL-c and HOMA-IR were significantly higher in the MAU patients than those in NMAU patients(P

Objective To investigate the application of two-factor theory of nursing students to improve the quality of teaching.Methods for nursing students of 2009,through the application of two-factor theory,to take effective measures to meet the health factors,and use incentives to improve nursing students to learn initiative and enthusiasm,for the nursing students of 2008,using the traditional clinical teaching methods.Results The 2008 nursing students theory test scores for (83.94 ±8.21 )points,operation test scores for (81.28 ±6.54)points,zongping scores for (82.47 ± 6.89 )points; The 2009 nursing students theory test scores for (87.17 ± 5.13 )points,operation test scores for (85.24 ± 4.61 )points,zongping scores for (86.27 ± 7.14)points,two groups of comparisons were statistically significant differences(t =-3.72,-4.14,-3.97;P ＜0.05).Conclusions Thetwo-factor theory can improve nursing students to learn initiative and enthusiasms to improve the quality of teaching.

In temporomandibular disorders (TMD), pain takes place when neuropeptides stimulate synovial tissue to produce several cytokines such as interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α, which activate neurons and glia of synovial membrane at the bilaminar regions of temporomandibular joint (TMJ). It has been reported that, after neurogenic differentiation, the synovial mesenchymal stem cells (SMSCs), deriving from TMJ, possess the same cytological features as the neuronal cells. This study examined the ability of substance P (SP) and calcitonin gene-related peptide (CGRP) to stimulate SMSCs and neurogenic SMSCs secreting inflammatory cytokines during TMD, evaluated the mutual effects of inflammatory cytokines and neuropeptides and tested the analgesic effect of hyaluronic acid (HA). The levels of IL-1β, IL-6 and TNF-α in SMSCs and neurogenic SMSCs in the presence of neuropeptides were measured by ELISA. SP and CGRP produced by SMSCs and neurogenic SMSCs were determined by RT-PCR and Western blotting. The results showed that the expression of SP and CGRP was significantly enhanced in the neurogenic SMSCs in response to IL-1β, IL-6 and TNF-α, and the effect was remarkably inhibited by HA. IL-1β, IL-6 and TNF-α, in return, could be enhanced in the neurogenic SMSCs upon stimulation by SP and CGRP. Neuropeptides and inflammatory cytokines might work mutually on the TMD pain. The HA-mediated analgesic effect may be implicated in the inhibition of SP and CGRP expression in neurogenic SMSCs.
Calcitonin Gene-Related Peptide ; biosynthesis ; Cells, Cultured ; Cytokines ; biosynthesis ; Female ; Humans ; Male ; Mesenchymal Stromal Cells ; metabolism ; pathology ; Substance P ; biosynthesis ; Synovial Membrane ; metabolism ; pathology ; Temporomandibular Joint ; metabolism ; pathology ; Temporomandibular Joint Disorders ; metabolism ; pathology

OBJECTIVETo study the protective effect of Naomaitong on inflammatory cascade response after focal cerebral ischemia reperfusion in aged rats.

METHODWe duplicated focal cerebral ischemia model with MCAO, with ischemia 3 h and I/R 1, 3, 6, 12 d points. The effect of Naomaitong on the nervous dysfunction score, the water content of cerebral constitution and the expression of TNF-alpha, VCAM-1, ICAM-1 and its mRNA were observed, and the group with nimodipine was as control.

RESULTThe nervous dysfunction score (I/R1, 3, 6 d), the water content of cerebral constitution (all the time points), the expression of TNF-alpha, VCAM-1 (I 3 h, I/R 1, 3, 6 d), ICAM-1 (I 3 h,I/R 1, 3, 6 d) and its Mrna (all the time points) in model group were higher than those of the sham-operated group; The nervous dysfunction score, the water content of cerebral constitution (I/R 3, 6, 12 d), the expression of TNF-alpha, VCAM-1 (I 3 h, I/R 1, 3 d), ICAM-1 and its mRNA (I 3 h, I/R 1, 3 d) in model group were decreased compared with that of model group. The nervous dysfunction score (I/R 6, 12 d), the expression of TNF-alpha, ICAM-1 (I/R 1d) and its mRNA (I/R 1, 3 d) in Naomaitong group were higher than that of Nimodipine group.

CONCLUSIONNaomaitong could protect brain cell from damage after focal cerebral ischemia reperfusion injury by inhibiting the expression of TNF-alpha, adhesion molecule.

Animals ; Brain ; metabolism ; pathology ; Brain Ischemia ; etiology ; metabolism ; pathology ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Infarction, Middle Cerebral Artery ; chemically induced ; Intercellular Adhesion Molecule-1 ; biosynthesis ; genetics ; Male ; Neuroprotective Agents ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; metabolism ; pathology ; Tumor Necrosis Factor-alpha ; metabolism ; Vascular Cell Adhesion Molecule-1 ; metabolism

Objective To develop a method for the simultaneous determination of 15mycotoxins in peanuts by ultra high performance liquid chromatography-tandem mass spectrometry with QuEChERS EMR-Lipid approach and stable isotope dilution. Methods The samples were extracted by 2% formic acid acetonitrile-water (50 : 50, V/V) and then purified with QuEChERS EMR-Lipid approach.The mycotoxins were fully separated on a pentafluorophenyl column under a gradient elution with methonal-0.01%formic acid aqueous solution.The mycotoxins were analyzed by UPLC-MS/MS with multiple reaction monitoring (MRM) mode and quantified by isotope internal standard method. Results Fifteen mycotoxins had good linear relationship in the certain correlation ranges with the correlation coefficients all above 0.995 and the detection limits were 0.1－10 μg/kg.The mean recoveries ranged from 81.2% to 115.3% with RSD (n=6) varying from 2.1% to 10.7%. Conclusion The method is simple, highly sensitive, practical, and proves to be suitable for quantitative analysis of 15 mycotoxins in peanuts.

OBJECTIVETo investigate the effect of high glucose on mitochondrial respiratory chain function in INS-1 cells.

METHODSThe pancreatic beta cell line INS-1 was divided into the normal control (NC), high glucose (HG), and N-acetyl-L-cysteine (NAC) pretreatment groups, which were cultured for 72 h in the presence of 5.5 mmol/L glucose, 16.7 mmol/L glucose, and 16.7 mmol/L glucose with 1.0 mmol/L NAC, respectively. The activities of the enzyme complexes I and III of the respiratory chain in the cells were assessed with spectrophotometry, the ATP levels were examined using a luciferinluciferase kit, and insulin levels detected by radioimmunoassay.

RESULTSThe activities of the respiratory chain enzyme complexes I and III were 1.53-/+0.24 and 1.08-/+0.22 micromol.mg(-1).min(-1) in high glucose group, respectively, significantly lower than those in the normal control group (2.31-/+0.33 and 1.92-/+0.39 micromol.mg(-1).min(-1), P<0.01). ATP and insulin levels also decreased significantly in high glucose group as compared with those in the normal control group (P<0.01). The addition of NAC partially inhibited high glucose-induced decreases in the enzyme complex activities, ATP levels and insulin secretion (P<0.05).

CONCLUSIONThe respiratory chain function is positively correlated to insulin secretion in INS-1 cells, and exposure to high glucose causes impairment of the two enzyme complexes activities through oxidative stress, resulting in the mitochondrial respiratory chain dysfunction. High glucose-induced damages of the mitochondrial respiratory chain function can be partially inhibited by NAC.

Cell Respiration ; drug effects ; Cells, Cultured ; Glucose ; pharmacology ; Humans ; Insulin-Secreting Cells ; cytology ; physiology ; Mitochondria ; physiology ; Oxidative Stress ; drug effects

OBJECTIVETo analyze the expression level of candidate tumor suppressor gene N-Myc downstream-regulated gene 2 (NDRG2) in human colon cancer.

METHODSThirty samples of colon cancer tissues with matched normal colon tissues were collected. The NDRG2 mRNA level was detected by semi-quantitive RT-PCR and the NDRG2 protein level was examined by Western blot and immunohistochemistry.

RESULTSTwelve samples of colon cancer tissues had low NDRG2 mRNA level and low protein level. The positive rates of NDRG2 in normal tissues and the tumorous colon tissues were 90.0%(27/30) and 53.3%(16/30) by immunohistochemistry respectively. There was a significant difference between two groups (P<0.05). The NDRG2 expression was not correlated with age, sex, metastasis of lymph node, depth of infiltration, as well as the Dukes staging(P>0.05), while it was correlated to the histology grading. The positive rate of NDRG2 in the well- and moderate-differentiation group was higher than that in the poor-differentiation group(P<0.05).

CONCLUSIONThe expression of NDRG2 is low in some colon cancer tissues, which indicates that the low level of NDRG2 expression may be engaged in the development of colon cancer.

Blotting, Western ; Colonic Neoplasms ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Proteins ; genetics ; metabolism

OBJECTIVETo identify and characterize mesenchymal stem cells from synovial membrane of temporomandibular joint in vitro.

METHODSSynovial mesenchymal stem cells (SMSCs) were obtained by limited dilution method and expanded in 25 ml flasks. Methyl thiazolyl tetrazolium (MTT) method was used to determine the cell growth cycles. The expressions of vimentin and keratin were respectively detected with immunocytochemistry, while the expressions of CD8, CD34, CD44, vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) were determined by flow cytometry.

RESULTSPure mesenchymal stem cells were of spindle shape and uniform in size, which were intensively positive in vimentin, but negative in keratin. The expression of CD44, VCAM-1 and ICAM-1 were also verified by flow cytometry.

CONCLUSIONSMesenchymal stem cells could be purified from adult synovial membrane of temporomandibular joint.

Cell Differentiation ; Cell Separation ; methods ; Cells, Cultured ; Flow Cytometry ; Humans ; Mesenchymal Stromal Cells ; cytology ; Synovial Membrane ; cytology ; Temporomandibular Joint