Objective To research the significanec of genes bel-2 and bax in chondrocyte apoptosis in experimental osteoarthritis in rabbits.Method Twelve New Zealand rabbits divided into two groups at random:model group and control group.Model group with osteoarthritis was duplicated by immobilizing with gypsum at extension position in right hind limb of rabbits.Rabbits of model group were executed after 6 weeks and chondrocytes were taken.Positive expression rate of bcl-2 and bax mRNA was measured by immunohisto- chemistry and Terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL).Results Positive expression rate of mRNA in bel-2 and bax of model group was obviously more than control group(P＜0.05).According to immunohistochemistry,and grey grade of positive expression of protein in bcl-2 and bax of chondrocytes in model group was lower and positive expression rate was gigher compared with control group (P＜0.05,P＜0.01).bcl-2 and bax of model group were lower than control group(P＜0.05).Conclusion Genes bcl-2 and bax participate and thus accelerate chondrocyte apoptosis of osteoarthritis together.

Background and purpose:EGFR-TKI (EGFR-tyrosine kinase inhibitors), represented by geiftinib and erlotinib, have exhibited signiifcant antiproliferative effects against non-small cell lung cancer (NSCLC) with low toxicity.EGFR gene mutation was discovered to be a predictive biomarker for EGFR-TKI treatment. Although the efifcacy of EGFR-TKI is limited toEGFR wild-type patients, it is still noticeable suggesting that some other mechanisms are responsible for it. The current study is aimed at evaluating the expression of phosphorylated EGFR in advanced NSCLC, investigating its relationship withEGFR mutations and EGFR-TKI efifcacy.Methods:EGFR gene mutations were detected by denaturing high performance liquid chromatography (DHPLC) in 205 stageⅢB-ⅣNSCLC patients. The expressions of phosphorylated tyrosine 1068 (pTyr1068) and 1173 (pTyr1173) were detected by immunohistochemistry.Results:The positive expressions of pTyr1068 and pTyr1173 were 80.0% (164/205) and 57.6% (95/165) respectively. None of them were related to clinical pathological characteristics (age, gender, pathological type, smoking status, disease stage).EGFR gene mutation rate was 44.9% (92/205), which was only related to smoking status (P=0.024) compared to other clinical pathological characteristics.EGFR gene mutations were poorly related to pTyr1068 expression (P<0.001) and not related to pTyr1173 expression (P=0.297). The objective response rate (ORR),disease control rate (DCR), and progressive free survival (PFS) of EGFR-TKI treatment in patients withEGFR mutations were 48.3% (43/89), 80.9% (72/89) and 8 months (95%CI: 6.11-11.42) respectively, which were signiifcantly higher than that ofEGFR wild-type patients [ORR=16.2% (17/105,P<0.001); DCR=56.2%(59/105,P<0.001); Median PFS: 2.1 months, (95%CI: 0.89-3.24;P=0.001)]. Superior ORR: DCR and PFS appeared in patients with pTyr1068 positive expression compared to negative [ORR: 37.7% (58/154)vs 5.0% (2/40,P<0.001); DCR: 74.7% (115/154)vs 40.0% (16/40,P<0.001); Median PFS: 7.0 monthsvs 1.2 months,P<0.001)]. Inversely, the patients with pTyr1173 positive expression had lower ORR, DCR and shorter PFS [ORR: 27.8% (25/90)vs 37.9%(25/66,P=0.123); DCR: 64.4% (58/90)vs 83.3% (55/66,P=0.007); Median PFS: 4.8 monthsvs 7.7 months (P=0.016)]. In subgroup ofEGFR wild-type patients, positive expression of pTyr1068 was 69.0% (69/100).EGFR wild-type patients with pTyr1068 positive expression had a prolonged PFS and elevated ORR and DCR compared to negative [median PFS: 3.6 monthsvs 1.2 months (P<0.001); ORR: 23.2%vs 3.2% (P=0.010); DCR: 69.6%vs 35.5% (P=0.001)]. Sixteen patients with pTyr1068 positive expression who responded to EGFR-TKI treatment in this subgroup had a remarkable PFS [median PFS: 15.6 months (95%CI:7.28-23.9)]. Multiple factor analysis showed that the expression of pTyr1068 was an independence predictor factor for EGFR-TKI treatment (OR=0.24, 95%CI: 0.16~0.37,P<0.001). Conclusion:Phosphorylation at Tyr1068 of EGFR might be a potential predictive factor for clinical response and survival of EGFR-TKI treatment in patients with advanced NSCLC, especially inEGFR wild-type patients.

Adolescent ; Adult ; Aged ; Antigens, CD ; biosynthesis ; Brain Neoplasms ; blood supply ; metabolism ; pathology ; Child ; Endoglin ; Female ; Glioma ; blood supply ; metabolism ; pathology ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; biosynthesis ; Immunohistochemistry ; Male ; Middle Aged ; Neovascularization, Pathologic ; metabolism ; pathology ; Receptors, Cell Surface ; biosynthesis ; Telomerase ; biosynthesis ; Young Adult

Chemical constituents of ethyl acetate extract of Sapium sebiferum leaves were isolated and purified by various chromatographic methods, including column chromatographies over silica gel, macroporous adsorption resin, and Sephadex LH-20, as well as preparative TLC and semi preparative HPLC. As a results, 15 compounds were separated from Sapium sebiferum leaves and their structures were examined by spectral analysis including NMR and MS data and identified as( + )-(7R,7'R,7"S,7'"S,8S,8'S,8"S,8'"S)-4", 4"'-dihydroxy-3,3',3",3',5,5'-hexamethoxy-7,9';7',9-diepoxy-4,8";4',8'"-bisoxy-8,8'-dineo-lignan-7",7"',9",9"'-tetraol(1) ,1-(4'- hydroxy-3'-methoxyphenyl)-2-[4"-(3-hydroxypropyl) -2", 6"-dimethoxyphenoxy] propane-1, 3-diol (2), Thero-2, 3-bis-(4-hydroxy-3- methoxypheyl)-3-methoxy-propanol(3) , threo-5-hydroxy-3,7-dimethoxyphenyl propane-8,9-diol (4), boropinol B (5), threo-8S-7-methoxysyringylglycerol(6), 5-hydroxymethylfurfural(7), 5-( methoxy-methyl)-1H-pyrrole-2-carbaldehyde (8), quercetin (9) , kaempferol (10), ethyl gallate(11), coniferaldehyde(12), vanillin(13), 7-hydroxy-6-methoxy-2H-1-henzopyran-2-one(14),and 1-heptacosanol (15). All compounds except for compounds 9-11,14 were separated from this plant for the first time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; Molecular Structure ; Plant Leaves ; chemistry ; Sapium ; chemistry

OBJECTIVETo compare effects of catgut embedding at "Zusanli" (ST 36) and "Shenshu" (BL 23) on Morphine analgesic tolerance and locomotor sensitization induced by chronic Morphine administration and the mechanism.

METHODSThe rats were randomly divided into a model group, a non-acupoint group, a Shenshu group and a Zusanli group. The rats, except those in the model group, were pretreated with acupoint catgut-embedding 10 days before the first Morphine injection. The Morphine-tolerance model was established and the pain threshold was detected by hot-plate test every day. Locomotor activities were recorded after the first Morphine injection and Morphine-challenging 1 week after withdrawal of Morphine. The positive neurons of nitric oxide synthetase (NOS) were showed by NADPH-d histochemical method.

RESULTSCompared with the non-acupoint group, catgut embedding at "Zusanli" (ST 36) could attenuate the Morphine analgesic tolerance and the increase of locomotor activities in rats. Meanwhile, the expression of NOS positive neurons in nucleus accumbens septi and dorsal striatum decreased in the Zusanli group. There were no significant differences between the Shenshu group and the non-acupoint group in the analgesic threshold and locomotor sensitization, but the expression of NOS positive neurons in the striatum region significantly decreased.

CONCLUSIONCatgut embedding at "Zusanli" (ST 36) can attenuate Morphine analgesic tolerance and reverse formation of locomotion sensitization induced by chronic Morphine administration, which are possibly related with inhibition of the expression of NOS positive neurons in nucleus accumbens septi and dorsal striatum.

Acupuncture Analgesia ; Acupuncture Points ; Analgesics, Opioid ; pharmacology ; Animals ; Catgut ; Drug Tolerance ; Male ; Medicine, Chinese Traditional ; Morphine ; pharmacology ; Motor Activity ; Rats ; Rats, Sprague-Dawley

Objective To develop a convenient method efficiently expands the frequency of specific CTLS.Methods We used different concentrations of CMV-speeific epitope peptides pp65 to stimulate PBMCs for expansion of CMV-specific CTLs.CMV-specifie CTLs were doubly labeled by tetramers-PE and CD_8-FITC for FACS analysis.Results The method expands CMV-speeific CTLs efficiently.CMV-specific CTLs were expanded from 1% to 20% of PBMCs quickly(namely 40% of CD_8~+ T cells).The method provided a large number of cells with tetramer staining of CD_8~+ T cells for FACS analysis from a single blood sampling.Conclusions Peptides stimulation methods are convenient,easy to operate and expanded CMV- specific CTLs efficiently.The increased frequencies of CMV-specific CTLs allowed the data of different individuals to be easily compared and sequentially evaluated.The methods lay the base for adoptive immunotherapy to prevent CMV disease.

To investigate the induction of apoptosis of mouse colonic adenocarcinoma CT26 cells by recombinant Clostridium difficile toxin B (rTcdB), CT26 cells were exposed to different concentrations of rTcd B. Inhibition of cell proliferation was measured by MTT assay. The activation of Caspase 3 was measured by colorimetric method. Cell morphological analysis and flow cytometry were performed to confirm cell apoptosis. rTcd B inhibited the proliferation of CT26 cells in a timeand dose-dependent manner. Caspase 3 activity in CT26 cells was elevated remarkably after rTcd B exposure for 6 h, 12 h, 18 h or 24 h, as compared with the control group. Morphological changes were observed by fluorescence microscopy. The exposure of rTcd B to CT26 cells induced a timeand dose-dependent apoptotic cell death as determined by flow cytometry analysis. The results showed that recombinant Clostridium difficile toxin B induced apoptosis of CT26 cells.

A stimulate method for determination of polybrominated diphenyl ethers ( PBDEs) and derivatives (OH-PBDEs and MeO-PBDEs), tetrabromobisphenol A (TBBPA), hexabromocyclododecane (HBCD) in egg samples was developed by gel permeation chromatography ( GPC) and dispersive solid phase extraction ( DSPE) combined with liquid chromatography tandem mass spectrometric ( HPLC-MS/MS) and gas chroma-tography-negative chemical ionization mass spectrometry ( GC-NCI/MS ) . The analytes were extracted with mixture of hexane and dichloromethane (1∶1, V/V) by accelerated solvent extraction (ASE), and purified by 100 mg C18 dispersive solid phase extraction ( SPE) sorbents followed with gel permeation chromatography (GPC) , and then analyzed by liquid chromatography tandem mass spectrometric (HPLC-MS/MS) and gas chromatography-negative chemical ionization mass spectrometry (GC-NCI/MS), respectively. The quantita-tion was carried out external standard method. The recoveries of objects were 64. 5%-97. 2% and 65. 6%-109 . 2% ( except BDE85 was 54 . 8%, OH-BDE-137 was 47 . 4%) spiked at 1 . 0 μg/kg or 5 . 0 μg/kg in egg white and egg yolk, respectively. The relative standard deviations (RSDs) were less than 20. 2%. The limits of quantitation (LOQ) for the object were 0. 01-0. 2 μg/kg.

Objective To explore the effect of different indexes on evaluating insulin resistance in obese children. Methods Oral glucose tolerance test (OGTT) was performed in obese children (n= 61) and age - matched normal volunteers( n= 23) Serum glucose and insulin levels were determined at 0,30,60,120,180 min after OGTT, insulin resistance was estimated by homeostasis model assessment of insulin resistance (HOMA-IR). Indexes such as the ratio of area under the curve of glucose(AUCG)/area under the curve of insulin(AUCI), the ratio of fasting blood sugar(FBG) and fasting blood insulin (FINS) were meanwhile calculated. Results The level of serum FINS was significantly higher in obese children(P

Background Eicosapentaenoic acid(EPA)function as the critical lipid mediators involved in several biological events in human body and play important role in suppressing the genesis of vascular endothelial growth factor (VEGF),migration and proliferation of vascular endothelial cells.Many ocular diseases were proved to be associated with neovascularization.Objecfive The purpose of this study was to investigate the inhibitory effect of EPA on the proliferation of human umbilical vein endothelial cells (HUVEC) indueed by VEGF. Methods HUVEC strain was cultured and passaged,and difierent concentrations of EPA were added to the medium with and without VEGF.The cultured cells were identified by antiofactor Ⅷ polyclonal antibody.The suppressing role of different concentrations of EPA on the proliferation of VEGF-induced or-uninduced HUVEC was assessed by MTT method.The influence of difierent concentrations of EPA on the cellular cycle of VEGF-induced HUVEC was assayed using flow eytometry.The expression of Flk-1,a receptor of VEGF,in the HUVEC Was detected by immunohistochemistry. Results Cultured HUVEC showed the ftlsiform in shape and presented with the cobblestone-like arrangement with the positive response for Ⅷ factor-related antigen.Various concentrations of EPA showed obviously inhibitory effect on VEGF-induced or-unindueed HUVEC at a dose-dependent manner (F=23.072.P=0.000).The inhibitory ability of EPA on VEGF-induced HUVEC was stronger than VEGF-uninduced HUVEC(F=41.417,P=0.000).In 24,48 and 72 hours,the action of EPA on the proliferation of HUVEC was gradually enhanced with the prolong of time(F=1.495,P=0.236).Cell cycle analysis indicated that EPA arrested VEGF-induced HUVEC in G0/G1 phase.The ratio of HUVEC in G0/G1 phase in EPA group was(75.83±1.56)%,and that in control groups was(68.62±1.44)%,showing a significant difference between them(t=-5.88,P=0.00),and no apoptosis of HUVEC was found in both groups.Flk-1 was strongly expressed in the cellular nucleus and cytoplasm in control group.However,the positive expressing intensity of Flk-1 in the HUVEC weakened,and the positive cell number was evidently less in EPA group. Conclusion EPA can inhibit the proliferation of VEGF induced HUVEC through arresting the synthesis of DNA of HUVEC and downregulate the expression of Flk-1 in HUVEC.These results suggest that EPA might exert an antiangiogenic effect.