AIM:To investigate the relationship of autophagy and reactive oxygen species ( ROS) in multiple myeloma cell line RPMI-8226 stimulated with doxorubicin.METHODS:The RPMI-8226 cells were stimulated with doxo-rubicin at different doses, and untreated cells were used as control.The protein expression of beclin 1 and LC3 was detec-ted by Western blot.ROS production was analyzed by DCFH-DA fluorescence staining.After treated with or without 3-methyladenine (3-MA), the ROS production and apoptosis in RPMI-8226 cells were determined by DCFH-DA and flow cy-tometry, respectively.After treated with or without antioxidants tempol and N-acetyl-L-cysteine ( NAC) , the expression of beclin 1 and LC3 in RPMI-8226 cells was determined by Western blot.RESULTS:The protein levels of beclin 1 and LC3Ⅱ/LC3Ⅰwere increased in the RPMI-8226 cells stimulated with doxorubicin compared with untreated group.The ROS production was increased in the RPMI-8226 cells stimulated with 2 mg/L doxorubicin compared with untreated group.After treated with 3-MA, the ROS production and apoptosis in the RPMI-8226 cells stimulated with doxorubicin were increased compared with doxorubicin group.After treated with antioxidant NAC or tempol, the expression of beclin 1 and LC3 II/I in the RPMI-8226 cells stimulated with doxorubicin was decreased compared with doxorubicin group.CONCLUSION:The autophagy and ROS levels are increased in RPMI-8226 cells stimulated with doxorubicin.Inhibition of autophagy increases the ROS production and apoptosis of RPMI-8226 cells stimulated with doxorubicin.Inhibition of ROS production reduces doxorubicin-induced autophagy in multiple myeloma cells.

Objective To evaluate the efficacy of acting κ opioid receptor for prevention of high altitude pulmonary edema (HAPE) in rats.Methods Forty male Sprague-Dawley rats,aged 8 weeks,weighing 250-300 g,were randomly divided into 5 groups (n =8 each) using a random number table:control group (group C),hypobaric hypoxia group (group H),normal saline + hypobaric hypoxia group (group NH),U50488H (a selective kappa-opioid receptor agonist) + hypobaric hypoxia group (group UH),and nor-binaltorphimine (norBNI,a selective kappa-opioid receptor antagonist) + U50488H + hypobaric hypoxia group (group NUH).The rats were put into the hyperbaric chamber and exposed to hypobaric hypoxia (atmospheric pressure 355 mmHg,partial pressure of oxygen 74 mmHg) for 2 days to induce HAPE.At 3 days before HAPE,normal saline 0.5 ml,U50488H 1.25 mg/kg,and nor-BNI 2.0 mg/kg were injected intraperitoneally once a day in NH,UH,and NUH groups,respectively,and in addition U50488H 1.25 mg/kg was injected intraperitoneally 10 min later in NUH group.After 2 h exposure to hypobaric hypoxia,mean pulmonary artery pressure (mPAP) was detected,and arterial blood samples were collected for determination of serum malondialdehyde (MDA) and erythropoietin (EPO) levels.The rats were then sacrificed and lungs were removed for microscopic examination and for determination of the levels of nitric oxide (NO),inducible nitric oxide synthase (iNOS),MDA,superoxide dismutase (SOD),endothelin-1 (ET-1),thromboxane B2 (TXB2),and 6-keto-prostaglandin F1α (6-keto-PGF1α) in lung tissues.Lung water content and TXB2/6-keto-PGF1α ratio was calculated.Results Compared with group C,mPAP,lung water content,ET-1,MDA,TXB2 and 6-keto-PGF1α levels,TXB2/6-ketoPGF1α ratio,and serum MDA and EPO levels were significantly increased,and iNOS,NO and SOD levels were decreased in the other four groups (P ＜ 0.05).Compared with group H,mPAP,lung water content,ET-1,MDA,TXB2 and 6-keto-PGF1α levels,TXB2/6-ketoPGF1α ratio and serum MDA and EPO levels were significantly decreased,and iNOS,NO and SOD levels were increased in UH group (P ＜ 0.05),and no significant changes were found in the indexes mentioned above in NH and NUH groups (P ＞ 0.05).The pathological changes of lung tissues were significantly attenuated in group UH as compared with H group.Conclusion Acting κ opioid receptor can produce prevention for HAPE in rats,and inhibition of lipid peroxidation and correction of the imbalance between vasoconstrictive factors and vasodilative factors may be involved in the mechanism.

Single-supervisor system is the widespread practice for graduates of clinical medicine at present. But in terms of education to clinical medical graduate students, students rotate in the departments of various disciplines for a long time, and there are more rotary department, resulting in some students' unbalanced learning development. Combined with the charac-teristics of the department of hematology, multi-supervisors training model was carried out to the rotating masters of other disciplines when they rotated in hematology department. With supervisor equipment and teaching implementation, we applied the principle of integrating theory with practice and trained students' innovation ability and doctor-patient communication ability. Compared with traditional single supervisor training pattern, multi-supervisors system can effec-tively improve the students' theoretical level, practical skills and comprehensive ability when the clinical medical graduate students rotate in the department of hematology.

Objective:To study the significance of LDL immune complexes in the pathogenesis of acute coronary syndrome(ACS).Methods:Enzyme linked immunosorbent Assay was used to measure the plasma LDL-IC in 139 patients with coronary heart disease(76ACS;63NACS) and in 111 control subjects.The patients with coronary heart disease were divided into multi-vessel diseased group(2-vessel diseased group and 2 vessels more)and mono-vessel diseased group(1-vessel diseased group)by coronary angiography.Results:①LDL-IC level was higher in patients with ACS than that in patients with non-ACS[(2.78?1.08)AU vs(1.74?0.45)AU,P

Objective:To study the significance of low-density lipoprotein immune complexes(LDL-IC) in the pathogenesis of complicated coronary lesion.Methods:Enzyme-linked immunosorbent assay(ELISA) was used to measure the serum LDL-IC in 139 patients with coronary heart disease and 111 normal controls.The patients were divided by coronary angiography into a multi-vessel diseased group,an ambi-vessel diseased group,mono-vessel diseased group;diffuse lesion group,a located lesion group,a serious stenoses group and a light stenoses group.Results: The LDL-IC level was significantly higher in the multi-vessel diseased group than in the ambi-vessel diseased group([2.75?1.22]AU vs [2.35?0.83]AU,P

Objective To investigate the diagnosis and treatment of post-transplant lymphoproliferative disease (PTLD) after hematopoietic stem cell transplantation (HSCT).Methods Clinical data of one case of PTLD,including clinical manifestations,diagnosis and treatment,were retrospectively analyzed.And the related literatures were reviewed.Results From January 2007 to November 2012,one case developed PTLD among 196 patients who were followed up after HSCT in the First Affiliated Hospital of Chongqing Medical University.The incidence of PTLD was 0.5％.Clinical manifestations of the patient were not typical,including frequent fever,multiple lymphadenopathy and multiple pulmonary nodules.The patient was diagnosed as diffuse large B-cell lymphoma (B-DLCL) by pathological examination of the swollen lymph node.After withdrawal of immunosuppressants,the temperature returned to normal,and lymphadenopathy and pulmonary nodules disappeared completely.Conclusion PTLD is a severe complication of HSCT with distinctive morphologic and clinical characteristics.The origin of this disease may be associated with Epstein-Barr virus infection and imrnunosuppressive therapy.It needs the pathologic detection to make a definite diagnosis.Several different treatment strategies have been employed,and reduction of immunosuppressive therapy may lead to regression of PTLD.

An HPLC method to determine six alkaloids of the Coptidis for Chinese Pharmacopoeia of 2015 Edition was established through C18 column. The mobile phase was CH3CN-0.25 mol·L-1 NH4Ac (36:64) (containing 8 mmol·L-1 SDS and adjusting pH 9.3 with ammonia) at the flow rate of 1 mL·min-1, the detective wavelength was 270 nm and the column temperature was 35 oC. The linear ranges of jatrorrhizine hydrochloride, columbamine hydrochloride, epiberberine hydrochloride, coptisine hydrochloride, palmatine hydrochloride and berberine hydrochloride were 0.006 96-0.233, 0.004 75-0.152, 0.003 30-0.528, 0.006 31-1.010, 0.004 71-0.753, 0.017 8-2.884 μg·mL-1, respectively. The average recoveries were 99.65%, 98.59%, 98.49%, 98.66%, 98.64%, 98.63% and RSD were 0.03%、0.15%、0.21%、0.12%、0.28%、0.23%, respectively. The method is simple and accurate, and can be used to determine the contents of jatrorrhizine, columbamine, epiberberine, coptisine, palmatine and berberine in the Coptidis.

AIM: Centrifuge training can improve forward acceleration (+Gz) endurance. This study was to analyzed the gene expression of rat heart affected by centrifuge, and to research the molecular mechanism of improving +Gz endurance by centrifuge training. METHODS: Differential expressed genes between high+Gz endurance (+16Gz) rats, of test group after trained 12 d and control were screened using suppression subtractive hybridization (SSH) and dot blot hybridization. The obtained expressed sequence tags (ESTs)were used as probes to perform RNA slot hybridization with heart total RNA isolated from each gruop of centrifuge training and high+Gz endurance and low+Gz endurance (+12Gz) rats, respectively. The positive ESTs were sequenced and analyzed using BLAST(nr) at NCBI.RESULTS: Three down-regulated ESTs were obtained from heart samples, all of them are new, and their expression levels were decreasing during centrifuge training. CONCLUSION: Centrifuge training can significantly affect the special gene expressions of rat heart, and the expression changes of these genes may be ralated to the mechainism that +Gz endurance can be improved by centrifuge training.

In our previous studies, DAZAP2 gene expression was down-regulated in untreated patients of multiple myeloma (MM). For better studying the structure and function of DAZAP2, a full-length cDNA was isolated from mononuclear cells of a normal human bone marrow, sequenced and deposited to Genbank (AY430097). This sequence has an identical ORF (open reading frame) as the NM_014764 from human testis and the D31767 from human cell line KG-1. Phylogenetic analysis and structure prediction reveal that DAZAP2 homologues are highly conserved throughout evolution and share a polyproline region and several potential SH2/SH3 binding sites. DAZAP2 occurs as a single-copy gene with a four-exon organization. We further noticed that the functional DAZAP2 gene is located on Chromosome 12 and its pseudogene gene is on Chromosome 2 with electronic location of human chromosome in Genbank, though no genetic abnormalities of MM have been reported on Chromosome 12. The ORF of human DAZAP2 encodes a 17-kDa protein, which is highly similar to mouse Prtb. The DAZAP2 protein is mainly localized in cytoplasm with a discrete pattern of punctuated distribution. DAZAP2 may associate with carcinogenesis of MM and participate in yet-to-be identified signaling pathways to regulate proliferation and differentiation of plasma cells.
Amino Acid Sequence ; Base Sequence ; Chromosomes, Human, Pair 12 ; genetics ; Chromosomes, Human, Pair 2 ; genetics ; Cytoplasm ; metabolism ; DNA Primers ; DNA, Complementary ; genetics ; Down-Regulation ; Gene Components ; Humans ; Likelihood Functions ; Models, Genetic ; Molecular Sequence Data ; Multiple Myeloma ; genetics ; metabolism ; Phylogeny ; Pseudogenes ; genetics ; RNA-Binding Proteins ; genetics ; metabolism ; Sequence Alignment ; Sequence Analysis, DNA

AIM: To explore the effect of pyrrolidine dithiocarbamate (PDTC), an NF-κB inhibitor, on the proliferation and apoptosis of human multiple myeloma U266 cells and its mechanisms.METHODS: The U266 cells were treated with PDTC at different concentrations (0, 25, 50, 100 and 200 μmol/L) in vitro.The growth inhibitory rate of the U266 cells was detected by CCK-8 assay and cell counting.The cell cycle of the U266 cells was determined by flow cyto-metry, and the apoptosis was examined by flow cytometry with Annexin V-FITC/PI staining.The effect of PDTC on the expression of DNA methyltransferase 1 (DNMT1) at mRNA and protein levels was measured by RT-qPCR and Western blot, respectively.The effects of PDTC on the protein levels of NF-κB (P65), DNMT1, Bcl-2, cyclin D1, cleaved caspase-3 and cleaved caspase-8 were determined by Western blot.RESULTS: The protein level of NF-κB (P65) was decreased after treatment with PDTC for 48 h or 72 h.PDTC inhibited the proliferation of U266 cells in both dose-and time-dependent manners.After treatment with PDTC for 48 h, the percentage of U266 cells in G2 phase increased compared with control group (P<0.05).PDTC induced the apoptosis of U266 cells in a dose-dependent manner.The expression of DNMT1 at mRNA and protein levels decreased (P<0.05).The results of Western blot showed that the expression of Bcl-2 in PDTC groups decreased, while the protein levels of cyclin D1, cleaved caspase-3 and cleaved caspase-8 were higher than those in control group (P<0.05).CONCLUSION: The NF-κB inhibitor PDTC inhibits the proliferation of U266 cells by inducing cell apoptosis.It may be related to the down-regulated expression of DNMT1, cell cycle arrest and activation of the apoptotic pathways.