Objective To explore the mechanism of apeptosis in rheumatoid arthritis synoviocyte induced by dihydroartemisinin. Methods Synovial tissues were cut from rheumatoid arthritis patients when who was under knee prosthesis. Apoptosis was detected with flow cytometry. Western blot was performed to assess ser473-phosphorylated Akt. EMSA (electrophoretic mobility shift assay) was used to ana-lyze NF-κB activation. Results Dihydroartemisin can induce apoptosis in rheumatoid arthritis synoviocyte in a dose-dependent manor from 2.5μmol/L to 10μmol/L. Rheumatoid Arthritis synoviocyte cultured with dihydroartemisinin in 5μmol/L or 10μmoL/L can significantly in-hibit serine 473 phosphorylation in Akt and activation of NF-κB. Conclusion Dihydroartemisinin can induce apoptosis in rheumatoid arthri-tis synoviocyte through Akt signal pathway.

Acute Disease ; Chromates ; poisoning ; Humans ; Male ; Occupational Diseases ; Young Adult

OBJECTIVE To investigate the ESBLs-producing isolates and their resistance and genotypes in Escherichia coli and Klebsiella in three hospitals of Wannan area.METHODS The MIC of 13 antimicrobials against 79 strains of E.coli and Klebsiella were determined by 2-fold agar dilution method.The confirmation test was used to detect ESBLs-producing strains according to CLSI 2005.PCR method was used to amplify beta-lactamases of ESBLs-producing isolates by 8 premiers,PCR products were purified and then sequencing was performed.RESULTS The ESBLs-producing rates in E.coli and Klebsiella in three hospitals of Wannan area were 61.5% and 45.0%,respectively.The resistance rates to CRO,CTX,CAZ,CIP and LEV of ESBLs-producing strains were 81.0%,81.0%,47.6%,71.4% and 50.0%,respectively,all isolates were susceptible to meropenem and imipenem.PCR indicated that CTX-M1,CTX-M13,TEM and OXA1 types of beta-lactamases were the four major genotypes of the three hospitals in Wannan area.CONCLUSIONS The isolation rates of ESBLs-producing E.coli and Klebsiella as well as their resistance rates to cephalosporins and quinolones in the three hospitals in Wannan area are quite high.Monitoring ESBLs-producing isolates should be strengthened in clinic and laboratory.

Objective To construct cDNA entry library and cDNA expression library of Armillifer agkistrodontis (A.) nymphs and make a preliminary immunoscreening for the cDNA expression library.Methods The nymphs were collected from the Kunming mice infected experimentally with A.agkistrodontis eggs and the total RNA were extracted from the nymphs using TRIzol Reagent.After purifying the mRNA,the synthesized cDNAs were cloned into the donor vector pDONR222 by BP reaction of Gateway technology and the recombinants were transformed into the DH10B cells by electroporation,the cDNA entry library was obtained.Next,the expression vector pDEST17 was ligated with entry clones by LR reaction,and the recombinants were transformed into the BL21 (DE3) cells.Hence,the cDNA expression library was constructed.Then,the expression library was immunoscreened with the mixed sera of mice infected with A.agkistrodontis,and the insertions of positive clones were sequenced.After that,the open reading frame(ORF) of positive slone sequence,the homology of the screened genes and their encoded proteins were analyzed by Finder and BLAST (basic local alignment search tool) program of National Center of Biotechnology Information(NCBI),and the discovered new genes were submitted into the GenBank.Besides,the physico-chemical properties,secondary structure and B cell epitopes of encoded proteins were also analyzed by bioinformatics software.Results The average titer and total clones of the cDNA entry library were 1.45 × 105 CFU/ml(colony-forming unit,CFU) and 1.74 × 106 CFU,respectively,and the range of fragment length of the inserted cDNA was between 0.2-4.0 kb,with an average of 1.4 kb.The total clones of cDNA expression library were 1.00 × 105 CFU,and the fragment length of the inserted cDNA was between 0.3-2.2 kb,with an average of 1.0 kb.Five positive clones,coded S1,S5,A1,D1 and F1,respectively,were obtained through preliminary immunoscreening.The sequence and homology of the five positive clones were sequenced and analyzed by BLAST program.No significant similarities were found in pentastomida species,which meant that they were all novel genes of A.agkistrodontis.The gene sequences were submitted to GenBank,with the accession number from JQ180451 to JQ180455.Also,results obtained by bioinformatics software showed that the predictive encoding proteins were all potential to be valuable recombinant diagnostic antigens.Conclusions The cDNA library of A.agkistrodontis nymphs is successfully constructed,and five new genes of A.agkistrodontis are discovered.The establishment of cDNA library and the discovery of the new genes will lay a foundation for further studying the gene functions and screening the immunodiagnostic antigens.

Adolescent ; Adult ; Dermatitis, Occupational ; etiology ; therapy ; Drug Eruptions ; etiology ; therapy ; Female ; Humans ; Male ; Methylprednisolone ; therapeutic use ; Trichloroethylene ; adverse effects

OBJECTIVETo study the protective effect of hyperoxia solution on acute lung injury caused by phosgene poisoning by observing the changes of PaO2 and malondialdehyde (MDA) contents, superoxide dismutase (SOD) activity in serum and Glutathione (GSH/GSSG) contents in lung tissues.

METHODSThe rabbits were divided into normal control group, hyperoxia solution (H0) and balance salt (BS) groups. Group HO and Group BS inhaled phosgene and the former was given intravenously hyperoxia solution (which was replaced by balance salt solution in Group BS). The content of MDA and the activity of SOD in serum were observed at different time points, the amount of GSH and GSSG in lung tissue were also measured.

RESULTS(1) The serum MDA contents increased and PaO2, SOD activity decreased significantly in Group HO and Group BS along with time increasing as compared with control group. The contents of GSH in lung tissue decreased in two groups compared with that in control group, however the contents of GSSG ascended instead. (2) At 3 and 8 h of the experiment, PaO2 of Group HO [(9.91 +/- 0.49), (9.15 +/- 0.46) mm Hg respectively] were significantly higher than those of Group BS [(9.03 +/- 0.76), (8.11 +/- 0.57) mm Hg respectively] (P < 0.01). The contents of MDA of Group HO (3.66 +/- 0.35), (5.31 +/- 0.15) micromol/L respectively] were lower than those of Group BS [(4.32 +/- 0.26), (7.4 +/- 0.33) micromol/L respectively] (P < 0.01). SOD activity in Group HO [(237.37 +/- 29.96), (208.10 +/- 18.80) NU/ml respectively] were higher than those of Group BS [(195.02 +/- 21.44), (144.87 +/- 21.26) NU/ml respectively] (P < 0.05 or P < 0.01). The content of GSSG lung tissue in Group HO (423.67 +/- 38.21) micromol/L were lower than those of Group BS (523.85 +/- 43.14) mol/L (P < 0.01). There were no significant differences in the content of GSH in lung tissues between Group HO and group BS.

CONCLUSIONHyperoxia solution can reduce acute lung injury of rabbits following phosgene poisoning.

Acute Lung Injury ; etiology ; metabolism ; pathology ; Animals ; Glutathione Peroxidase ; metabolism ; Hyperoxia ; Lung ; drug effects ; metabolism ; pathology ; Malondialdehyde ; analysis ; Oxygen ; administration & dosage ; pharmacology ; Phosgene ; poisoning ; Rabbits ; Superoxide Dismutase ; metabolism

OBJECTIVETo investigate the relation between the progressive effects of chronic intermittent hypoxia (CIH) on cognitive function and the change of cholinergic neuron.

METHODSForty adult male Sprague-Dawley rats were randomly averagely divided into four groups: control group, CIH 1 week group, CIH 3 week group and CIH 5 week group. The cognitive function was assessed by the Morris Water Maze. The necrosis neurons in prefrontal cortex and hippocampus were observed and counted. The cholin acetyltransferase (ChAT) immunostained cells in prefrontal cortex and hippocampus were identified and quantitated.

RESULTSThe spatial learning and memory impairments progressed from 1 to 5 5 weeks in rats. Compared with the control group, the cognitive impairments in CIH5w group were significant (P < 0.05). The degeneration or necrosis neurons in prefrontal cortex and hippocampus were significantly increased in CIH rats, and worsen gradually along with the hypoxia. The ChAT immunostained cells in prefrontal cortex and hippocampus were gradually reduced. The ChAT immunostained cells of prefrontal cortex and hippocampus in CIH3w group and CIH5w group were less than that in control group (P < 0.05).

CONCLUSIONChronic intermittent hypoxia induced slowly progressive spatial learning and memory impairments in rats, which maybe associated with the damage of neurons and the reduction of ChAT in prefrontal cortex and hippocampus.

Animals ; Cholinergic Fibers ; pathology ; physiology ; Cholinergic Neurons ; pathology ; physiology ; Cognition ; physiology ; Hippocampus ; cytology ; physiopathology ; Hypoxia ; physiopathology ; Male ; Maze Learning ; physiology ; Memory Disorders ; etiology ; physiopathology ; Prefrontal Cortex ; cytology ; physiopathology ; Rats ; Rats, Sprague-Dawley

Alcoholism ; metabolism ; Animals ; Brain ; metabolism ; Chemokine CCL2 ; genetics ; metabolism ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, CCR2 ; genetics ; metabolism

Alcoholism ; physiopathology ; prevention & control ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Glucosides ; pharmacology ; Male ; Maze Learning ; drug effects ; Memory Disorders ; prevention & control ; Prefrontal Cortex ; metabolism ; physiopathology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; genetics ; metabolism ; Stilbenes ; pharmacology

Background Some drugs with inhibitory effect on the proliferation of lens epithelial cells have a limiting application in clinic because of their adverse response.To screen the effective and less side-effect drug for supressing LECs growth is very inportant for the prevention and treatment of after cataract.Objective This study was to explore the effects of docetaxel on LECs growth and compare its role with epirubicin hydrochloride,pirarubicin hydrochloTide and rahitrexed.Methotis Immortalized human LECs line (SRA01/04) were cultured and passaged.Different concentrations of docetaxel,epirubicin hydrochloride,pirarubicin hydrochloride and rahitrexed were added into the medium respectively for 24.48 and 72 hours.The proliferation of LECs was detect by M1Yr.Flow cytometry analysis Was used to analyze the influence of different concentrations of docetaxel on cellular cycle at 48 hours after addition of docetaxel,and Annexin V-FITC/PI marking method was used to assesse the apoptosis of LECs under the action of docetaxel.Expression of bcl-2 protein in LECs Was evaluated by Westeru blot. Result The growth rate of LECs Wag 100%in 8-519 pmol/L doeetaxel groups with the normal cell shape.Majority of abnormal cells and low growth rate were found in 66 nmoVL docetaxel group at 48 and 72 hours.The IC50 of docetaxel was lowest in 48 and 72 hours in docetaxel group in comparison to epirubicin hydrochloride and pirarubicin hydrochloride. However,no evident inhibition on LECs growth in 23.22-523.56 μmol/L of raltitrexed.At 48 hours,the percentage of LECs in G2/M phase increased as the asccnte of concentration of docetaxel,showing a significant difference among 4 groups(F=2633.05,P＜0.01).The percentage of early apoptotic cells increased to 22.4%(χ2=20.00,P＜0.01) and 27.9%(χ2=42.68,P＜0.01)from normal control 3.1% at 48 hours after LECs exposed to 8.3 nmol/L and 266 nmol/L docetaxe.The expression of bcl-2 protein in LECs was obviously weakened after addition of docetaxel,especially 8.3 nmol/L docetaxel group. Conclusion Docetaxel,epirubicin hydrochloride and pirarubicin hydrochloride can inhibit the proliferation of human LECs in vitro.But there is no supression on LECs growth inraltitrexed.Docetaxel is proved to have a strongly arrested effect on the proliferation of LECs in comparison with epirubicin hydrochloride and pirarubicin hydrochloride and play its role at concentration-and time-dependent manner.