AIM:To investigate the effects of MG132,one of the proteasomal peptide aldehydes inhibitors,on lipopolysaccharide(LPS)-induced nuclear transcription factor-kappa B(NF-?B) activation,the production of nitric oxide(NO) and tumor necrosis factor-alpha(TNF-?),as well as the expression of inducible nitric oxide synthase(iNOS) in murine macrophage line RAW264.7.METHODS:Reporter gene assay was used to examine the activity of NF-?B by pNiFty-SEAP/HEK293 cells,which were transfected with the pNiFty reporter plasmid into human embryo kidney cells(HEK293).Fluorescence substrate DAF-2DA was used to testify NO level in Raw264.7 cell line induced by LPS.Furthermore,the secretion of TNF-? was examined by ELISA.Western blotting was used to reveal the expression of iNOS and I?B-?.RESULTS:MG132 significantly decreased the secretion of TNF-? induced by LPS,with the inhibitory rates of 36.7% and 60.4% to 5 ?mol/L and 10 ?mol/L MG132,respectively.The pro-inflammatory mediator NO production was decreased in a dose-dependent manner with the inhibitory rates increasing from 29.5%(2 ?mol/L) to 55.9%(10 ?mol/L).Pretreatment with MG132 reduced the expression of iNOS,but restored the I?B restrain caused by LPS treatment.Observed by a reporter gene assay,TNF-?-induced NF-?B activity was decreased gradually by addition of increasing concentration of MG132(2.5-10 ?mol/L).CONCLUSION:Our results suggest an anti-inflammation effect of MG132 by the suppression of LPS-induced the production of pro-inflammatory mediators including NO and TNF-?,and the expression of iNOS,which probably mediates the blockage of I?B degradation and NF-?B activation.

Many problems and challenges,which are closely related to the background of multiculture,exist in the current education of humanistic quality for medical students in China.Therefore,it is important to enhance the humanistic quality of medical students under the multi-cultural background.Following countermeasures can be taken to strengthen the humanistic quality of medical students,including establishing the curriculum for humanistic education,enriching humanistic knowledge of medical students,building dense on-campus cultural environment,transforming students′ recognition of humanistic education,and developing various social practices of humanistic quality.

It has been well known that apoptosis induction and cell cycle arrest are typical biological effects observed in cancer cells after proteasome inhibition. TH2 is a new natural xanthone analogue isolated from the resin of Garcinia hurburyi tree. Here, the cell growth inhibition of TH2 on human hepatocellular carcinoma cell line (Bel-7402) was evaluated in vitro using SRB assay. The treatment of 10 ?mol/L TH2 reduced the surviving fraction from 86% (12 h) to 17.2% (48 h). To assess whether TH2 induce apoptosis, the appearance of sub-G1 peak, a specific fraction for apoptosis was detected by flow cytometry analysis. Progressive increase in the percentage of apoptotic population was observed in a dose-and time-dependent manner. Furthermore, a cleavage of poly (ADP-ribose) polymerase (PARP), a marker of early apoptosis, was observed clearly when the cells exposed to 10 ?mol/L of TH2 for 24 h by immunoblotting analysis. In vitro activities of 20 S proteasome purified from human erythrocytes on fluorogenic peptide substrates revealed that TH2 inhibited the trypsin-like, chymotrypsin-like and peptidylglutamyl peptide hydrolyzing activities in dose-dependent manner. Moreover, the turnover of tumor suppressor p53, a sign of deregulation of cell cycle progression and apoptosis induction by classical proteasome inhibitors, was disrupted in Bel-7402 cells. All these data indicate that TH2 had inhibitory effect on the proliferation of Bel-7402 cells and induction of apoptosis, which might be related to its inhibition of proteasome.

Objective To investigate the protective effects of 1-hydroxy-2, 3, 5-trimethoxyxanthone (QGS) on acute lung injury of mice induced by ip lipopolysaccharide (LPS). Methods Mice were pretreated with QGS for 7 d. Murine models of acute lung injury were duplicated by injection of LPS 20 mg/kg intraperitoneally. In 12 h, the lung weight index was observed and the NO level in the bronchoalveolar lavage fluid (BALF) was measured with kits. The lung was also assessd for the expression of I-?B, inducible nitric oxide synthase (iNOS), and cyclooxygenase-Ⅱ (COX-2) using Western blotting analysis. Lung pathological changes were also observed by HE in each group. Results The lung weight index of injury lung in mice induced by LPS was decreased in 500 mg/kg QGS group (P

Objective To investigate the accuracy of functional magnetic resonance imaging(fMRI) and contrast-enhanced ultrasound on evaluation of the breast cancer patients after neoadjuvant chemotherapy.Methods Forty-eight patients with breast cancer who were confirmed by biopsy,undergoing 4 cycles weekly dose-concentrated programs (wPC) neoadjuvant chemotherapy,evaluated the efficacy by fMRI and contrastenhanced ultrasound.Results In pathology,complete remission was 12 cases (25.0％,12/48),non remission was 13 cases (27.1％,13/48).In contrast-enhanced ultrasound,complete remission was 20 cases (41.7％,20/48),non remission was 10 cases (20.8％,10/48).In fMRI,complete remission was 17 cases (35.4％,17/48),non remission was 11 cases (22.9％,11/48).The P values more than 0.05 showed no statistically significant among three methods,the Kappa test showed that fMRI were superior to contrastenhanced ultrasound on evaluating the efficacy of neoadjuvant chemotherapy.Conclusions fMRI and contrast-enhanced ultrasound can be used in evaluating the efficacy of neoadjuvant chemotherapy,however,the data shows fMRI is superior to contrast-enhanced ultrasound.Contrast-enhanced ultrasound must be as the supplementary method.

ObjectiveTo explore the effects of different doses of PcTx1,a specific blocker of acid-sensing ion channel 1a,on global cerebral ischemia/repedfusion (I/R) injury in rats,MethodsSixty adult male Sprague Dawley rats (weighing 250-300 g) were randomly divided into 6 grups ( n =10 each):sham operation group (group S),I/R group,different doses of PcTx1 ( 10 ng/ml,group P1 ; 25 ng/ml,group P2 ; 50 ng/ml,group P3 ;and 500 ng/ml,group P4 ) groups.Global cerebral ischemia was induced by the modified procedure of Pulsinelli 4-vessel occlusion.In groups P1,P2,P3 and P4,different doses of PcTx1 ( 10,25,50 and 500 ng/ml),6 μl each,were respectively injected into the lateral cerebral ventricle at the initiation of reperfusion,while equal volume of double distilled water was injected instead in group I/R.Six rats in each group were sacrificed at 24 h of reperfusion,and the brains were immediately removed,Thereafter,the contents of malondialdehyde (MDA),reduced glutathione (GSH) and ritric oxide (NO),the activities of constitutive NO synthase (eNOS) snd inducible NO synthase (iNOS) were detected in hippocampus.Four rats in each group were sacrificed at 72 h of reperfusion,and hematoxylin and eosin staining was used to observe the pathomorphological changes of the hippocampal neurons.ResultsCompared with group S,the other groups showed decreases in the contents of GSH,while increases in the contents of MDA and NO and the activities of cNOS and iNOS ( P ＜ 0.05 or 0.01 ).The contents of GSH increased,while the contents of MDA and NO and the activities of cNOS and iNOS decreased in groups P2,P3 and P4 compared with group I/R ( P ＜ 0.05 or 0.01).Compared with group P1,the contents of GSH increased,the contents of MDA and the activities of cNOS decreased in groups P2,P3 and P4,and the contents of NO and the activities of iNOS decreased in groups P3 and P4 ( P ＜ 0.05 or 0.01 ).Compared with group P2,the activities of iNOS decreased in groups P3 and P4(P ＜ 0.05 or 0.01).The damage to neurons in hippocampal CAI was severe in groups I/R and P1,but it was attenuated in groups P3 and P4.ConclusionPcTx1 25,50 and 500 ng/ml (6 μl)injected into lateral cerebral ventricle can attenuate global cerebral I/R injury in rats,and the dose 50 ng/ml (6 μl) is more suitable.

A high performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) method was developed for determining degradation dynamics and final residues of myclobutanil 25% WG in ginseng root, stem, leaf and soil. The samples were extracted with acetonitrile, cleaned-up with primary secondary amine (PSA) solid phase extraction cartridge, separated by Kromasil Eternity-5-C18 (2.1 mm x 150 mm, 5 microm) column with a gradient of acetonitrile and 0.1% formate in water as mobile phases, and analyzed with the multiple reaction monitoring (MRM) in positive ion mode by employing the external standard method. The average recoveries and the relative standard derivations (RSDs) of myclobutanil at the spiked level of 0.01-0.20 mg x kg(-1) were 80.9%-90.7% and 5.54%-9.29%, respectively, and the limit of quantification (LOQ) was 0.005 mg x kg(-1). The method with good reproducible, high precision and low detection limit could meet the requirements of residual analysis on ginseng production. The half-lives of myclobutanil were from 6.25 days to 9.94 days in ginseng root, stem, leaf and soil at spraying dosage of 1 152 g x hm(-2) The final residues were below 0.060 1 mg x kg(-1) in root, below 0.081 7 mg x kg(-1) in stem, 0.006 0-0.102 2 mg x kg(-1) in leaf and below 0.037 6 mg x kg(-1) in soil at spraying dosage range from 576 to 1 152 g x hm(-2). It is recommended that the MRLs of myclobutanil in dried ginseng may be suggested to be 0.10 mg x kg(-1) temporarily, and the preharvest interval was set at 35 days.
Chromatography, High Pressure Liquid ; methods ; Drug Contamination ; Fungicides, Industrial ; chemistry ; Nitriles ; chemistry ; Panax ; chemistry ; Plant Leaves ; chemistry ; Plant Roots ; chemistry ; Plant Stems ; chemistry ; Soil Pollutants ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods ; Tandem Mass Spectrometry ; methods ; Triazoles ; chemistry

Objective To study influencing factors for giving up chemotherapy during initial treatment in elderly patients with multiple myeloma.Methods A total of 156 elderly patients diagnosed as multiple myeloma from 2000-2010 were retrospectively analyzed.And 123 young patients with multiple myeloma were selected as control group.The causes for giving up the chemotherapy and its related factors were recorded and analyzed.Results The rate of giving up the initial chemotherapy was higher in the elderly than in young patients [58.3％ (91/156) vs.35.0％ (43/123),P＜0.05].51.7％ of the elderly patients did not accept chemotherapy after diagnosis.The main factors for giving up the initial chemotherapy in elderly patients were critically ill,economic difficult and the low tolerance.However,the economic difficulty was the first cause in the young group.Conclusions The rate of giving up the chemotherapy in the elderly patients is high.Old age,severe conditions,economic difficulty,and low tolerance are the independent factors.

Objective To investigate the role of acid-sensing ion channel 1a(ASIC1a) in global cerebral ischemia-reperfusion injury in rats.Methods Forty male SD rats weighing 250-300 g were randomly divided into 4 groups (n =10 each): sham operation group (group S),cerebral ischemia-reperfusion group (group I/R),solvent control group (group SC) and group PcTX1 (a ASIC1 a blocker,group P).Global cerebral ischemia-reperfusion was induced by four-vessel occlusion.PcTX1(500 ng/ml)6 μl or solvent 6 μl was injected into the crerbral ventricular at the begining of reperfusion in groups P and SC respectively.The rats were sacrificed at 24 h of reperfusion,and then the hippocampi were removed for determination of Caspase-3,Bcl-2 and Bax protein expression and microscopic examination.Results Compared with group S,the expression of Caspase-3,Bcl-2 and Bax protein was up-regulated in groups I/R,SC and P (P ＜ 0.05).Compared with group I/R,the expression of Caspase-3 and Bax was down-regulated,and the expression of Bcl-2 was up-regulated in group P ( P ＜ 0.05).There was no significant difference in Caspase-3,Bcl-2 and Bax protein expression between groups I/R and SC (P ＞ 0.05).The histopathologic damage was ameliorated in group P as compared with group I/R.Conclusion ASIC1a can induce global cerebral ischemia-reperfusion injury in rats by up-regulating Caspase-3 and Bax expression,and down-regulating Bcl-2 expression and inducing apoptosis.

Objective To investigate the expression and the significance of toll-like receptor 3 (TLR-3)in placenta,tumor necrosis factor-α(TNF-α)in maternal and cord blood of idiopathic fetal growth restriction(IFGR),and their correlation with the pathogenesis of symmetric and asymmetric IFGR.Methods From April 2008 to April 2009,42 primiparae of singleton pregnancy and their IFGR babies,who delivered at term through cesarean section, in the Third Affiliated Hospital of Zhengzhou University were enrolled. All subjectects were divided into symmetric IFGR group (n=20) and asymmetric IFGR group (n =22). Another 42 non-IFGR pairs were randomly selected as the control group. The polink-2 plus polymerized horseradish peroxidase (HRP) immunohistochemical method and the enzyme linked immunosorbent assay (ELISA) were applied to detect TLR-3 and TNF-α levels. Results (1) The expression of TLR-3 protein were observed in all maternal placenta of the three groups. TLR-3 essentially expressed in syncytiotrophoblasts and hofbouer cells in the symmetric IFGR and control group, but expressed mostly in hofbouer cells and less in syneytiotrophoblasts in the asymmetric IFGR group. (2) The expression of TLR-3 in the syncytiotrophoblasts of the symmetric and asymmetric IFGR group was significantly lower than in the control group (111±14 and 118±11 vs. 156 ± 9, P<0. 01). The number of TLR-3 positive in Hofbourer cell in the symmetric IFGR group was lower than the control group (8. 9±2. 8 vs 17.5±2. 8, P <0. 01 ), but the number in the asymmetric IFGR group was higher (23.8±3.7) compared with the control group (P <0. 01). (3) The TNF-α levels in the maternal and cord blood of the symmetric and the asymmetric group were higher than that of the control group [maternal : (90±10) μg/L and ( 86±11 ) μg/L vs. (73±9) μg/L;cord blood: (92±12) μg/L and (96±8) μg/L vs. (79±9) μg/L;P<0.01]. (4) Neither symmetric nor the asymmetric IFGR group showed any correlations between the maternal and cord blood levels of TNF-α (P>0. 05). (5) Significant correlation was found between the TNF-α level of the cord blood and TLR-3 expression in the placenta in both the symmetric and asymmetric IFGR group(P<0. 05),but no relationship was found between the maternal blood TNF-α level and TLR-3 expression in the placenta (P>0. 05). Conclusions The variantions of TLR-3 expression in placenta and the increased expression of TNF-α in cord blood are associated with the genesis IFGR. The reduced expression of TLR-3 may related to symmetric IFGR, while the increased TLR-3 level in hofbouer cells may lead to asymmetric IFGR.