OBJECTIVETo establish the comparative proteomic profiles of retinoid acid (RA) resistant human acute promyelocytic leukemia (APL) NB4-R1 cells before and after apoptosis induced by realgar (tetra-arsenic tetra-sulfide, As4S4).

METHODSFirst a serial of assays were performed using MTT, transmission electron microscopy, Annexin V FITC/PI double-stain, flow cytometry and confocal laser scanning microscopy to qualitatively and quantitatively observe the in vitro apoptosis inducing effect of realgar on RA-resistant cells. Then the comparative proteomic profile before and after NB4-R1 apoptosis was established using high-resolution two-dimensional electrophoresis system.

RESULTSThe inhibition effect of realgar on NB4-R1 cell growth was dose and time dependent. The 24-h 50% inhibiting concentration (IC50) was 24.06 +/- 0.19 micromol/L, and the 48-h IC50 9.50 +/- 0.13 micromol/L, and 72-h IC50 6.55 +/- 0.03 micromol/L, respectively. 24 h and 48 h were the early and late phase of major NB4-R1 apoptotic cell populations induced by 25 micromol/L realgar respectively. Differential proteomic profiles before and after realgar induced NB4-R1 apoptosis were successfully established. Averagely 1069, 975 and 893 spots could be detected of the untreated group (R0), the 24-h treatment group (R24), and the 48-h treatment group (R48), respectively by ImageMaster 2D Platinum Software. The matching rate between R24 and R0 was 79.94% and that between R48 and R0 69.33%, and that between R24 and R48 71.91%.

CONCLUSIONDifferential proteomic profiles of realgar induced NB4-R1 apoptosis were successfully established for the first time, which provided a basis for comprehensively understanding the signal transduction of realgar induced apoptosis in RA-resistant APL cells, also for screening new bio-markers and drug targets of hematopoietic malignant tumor.

Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; drug effects ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Leukemia, Promyelocytic, Acute ; pathology ; Proteome ; analysis ; Sulfides ; pharmacology ; Tretinoin ; pharmacology

Objective To evaluate the efficacy of first-line CHOP or CHOP-like regimen on patients with advanced staged angioimmunoblastic T-cell lymphoma (AITL).Methods Between Aug 2006 and Sep 2014,twenty-nine AITL patients who were newly diagnosed without prior treatment were included in study.The clinical features,efficacy and survival were analyzed retrospectively.Results Median age of these patients was 59 years old.All patients had stage Ⅲ/Ⅳ disease.17 (58.6 ％) cases presented with B symptoms.26 (89.7 ％) cases had an international prognostic index (IPI) score ≥2,and 20 (69.0 ％) cases had elevated LDH,9 (31.0 ％) cases had ≥2 extranodal involvements.The median follow-up time was 20 months.Overall response rate was 69.0 ％ (20/29).Five (17.2 ％) patients achieved complete remission (CR+CRu),15 (51.7 ％) patients achieved partial remission,and 3 (10.3 ％) patients had stable disease (SD),6 (20.7 ％) patients had progressive diseases(PD).Median progression-free survival (PFS) was 6 months.1-and 2-year PFS rates were 39.0 ％ and 20.0 ％.1-,2-and 5-year overall survival (OS) rates for all patients were 76.8 ％,53.4 ％ and 17.1％,respectively.PFS was significantly better in chemotherapy-sensitive patients (P ＜ 0.001).The responses to chemotherapy had a tendency of affecting the OS,but it failed to reach statistical significance (P ＞ 0.05).Conclusions The CHOP or CHOP-like regimen maybe induce unfavorable efficacy in AITL patients.Further therapeutic options are required to improve the outcome.

Objective: To study the antibacterial activity in vitro of Xiyanping injection combined with cefoxitin, cefuroxime, ceftriaxone, cefmenoxime, amikacin, levofloxacin and meropenem against staphylococcus aureus, escherichia coli, klebsiella pneumoniae, pseudomonas aeruginosa and acinetobacter bauman.Methods: Minimum inhibitory concentration (MIC) of the 7 kinds of antimicrobial agents used individually or combined with Xiyanping injection on germs were determined respectively on 96 plates by using the agar dilution method and the checkerboard method.Fractional inhibitory concentration index (FICI) was calculated to evaluate the antibacterial activity of Xiyanping injection combined with the 7 kinds of antimicrobial agents in vitro.Results: MIC of Xiyanping injection combined with cefoxitin was the same as that of cefoxitin used individually on escherichia coli.MIC of levofloxacin combined with Xiyanping injection showed no difference with that of levofloxacin used individually on klebsiella pneumoniae.Besides, Xiyanping injection could increase the MIC of the other antimicrobial agents on germs.Conclusion: Xiyanping injection combined with antimicrobial agents on staphylococcus aureus, escherichia coli, klebsiella pneumoniae shows antagonistic and irrelevant relationship.

Objective To determine the usefulness of real time three-dimensional transesophageal echocardiography(RT-3D TEE) in the preoperative assessment of mitral valve (MV) pathology by comparing images with surgical findings and to evaluate the function of MV postoperatively. Methods Nineteen consecutive adult patients with established diagnosis of mitral regurgitation(MR) scheduled for surgical correction were enrolled. Intraoperative 2D and 3D transesophageal echocardiography(TEE) were performed. All the 3D images were compared with findings obtained from direct surgical inspection. Postoperative RT-3D TEE was performed immediately to evaluate outcomes of mitral annuloplasty. Results Superb 3D-TEE en face views of the MV were obtained in all patients. Correct diagnoses of MV pathology in agreement with the surgical findings were made in 16 of 19 patients (84. 2%), however in the remaining 3 patients, the diagnoses were incorrect despite good image quality. In one, RT-3D TEE diagnosed prolapse of A2 segment and P2 scallop,which was not confirmed at surgery, but MV degeneration and annular dilatation were observed. In another one, RT-3D TEE revealed prolapse of P3 scallop, while at surgery prolapse involved P2 and P3 with ruptured chordae. In the rest one,surgically detected prolapse of P2 was missed by RT-3D TEE. On the other hand with 2D TEE,the diagnoses correlated poorly with surgical findings,only 10 patients were accurately diagnosed (52.6%). In some patients,2D TEE was able to identify the MVP, but it had difficulty in defining the exact location of the prolapsed segment or scallop. The severity of MR decreased significantly after surgery. MV repair was successful in all patients except one(5.1%), in whom moderate to severe MR was still present and MV replacement was conducted subsequently as an alternative.Conclusions RT-3D TEE provides excellent imaging of MV components, including the anterior and posterior leaflets, as well as annulus and subvalvular structures, which can be especially helpful in planning the most appropriate surgery strategy. RT-3D TEE offers exact anatomic characteristics of mitral annuloplasty rings and bands, providing additional information for the evaluation of surgical outcomes.

OBJECTIVETo explore whether the constructed vector of short haprin in vivo can induce human glioma cell line BT325 to produce RNAi duplexes and reverse the expression of MDR1 gene.

METHODSThree 62nt oligonucleotide fragments (shRNA) were constructed according to GenBank MDR1 sequence and were cloned to the retrovirus-delivered vectors. After transfected these vectors directly into the human malignant glioma BT325 cells by lipofectamine 2000 with enhanced green fluorescence protein (EGFP) co-transfecting, the MDR1 gene silence effects were detected by the changing level of mRNA and P-glycoprotein including real time PCR (RT-PCR), Northern blot and Western blot analysis. To assess the multidrug resistance against adriamycin (ADR) and VCR, cell proliferation assays were performed by cell counting kit-8.

RESULTSThe RNAi plasmid vectors were constructed successfully. RT-PCR showed MDR1 mRNA was significantly reduced (P < 0.05). Northern blot analysis showed that the gene silence became most intense at 48 hours after transfection. Western blot analysis demonstrated that P-gp expression was reduced at different time to 12.9%, 30.3% and 4.8%, respectively. The chemosensitivity assays indicated that the transfected cells showed an enhanced sensitivity to ADR and VCR. Based on the value of IC(50), BT325 cells had significantly increased sensitivity to the drugs.

CONCLUSIONThe sequence specific RNAi can inhibit MDR1 mRNA and P-gp expression in the glioma cell line. It may reverse multidrug resistance phenotype, therefore, may provide promising therapeutic modalities in the treatment of human glioma.

ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; genetics ; Antibiotics, Antineoplastic ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Glioma ; metabolism ; pathology ; Humans ; Plasmids ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; pharmacology ; Transfection ; Vincristine ; pharmacology

Objective To explore the different apoptotic gene expressions and apoptotic signaling transduction of human rhabdomyosarcoma (RD) cells infected by enterovirus 71 (EV71) in different stage.Methods The survival of EV71-infected RD cells was observed by trypan blue staining.The apoptotic morphology and rates of RD cells were surveyed and detected by Annexin V-FITC/PI staining and flow cytometry,respectively.PCR array was employed to analyze 88 apoptotic gene expressions from EV71-infected RD cells at 8 h and 20 h postinfection (p.i),respectively.Results After RD cells were infected with EV71 (MOI =5) at 8 h p.i,the viability was significantly decreased.Flow cytometry data demonstrated that the apoptotic rates of EV71-infected RD cells had increased to 18.0％ and 19.1％ at 20 h p.i in early and later stage respectively.RT-PCR array studies revealed significant variations in the expression of apoptotic genes.Among 88 genes,only the expression of IFN-β1 was upregulated 5.22 folds,whereas 47 genes including ACIN1,Akt,Apaf1,caspase and CIDEB were found to be downregulated that were lower than 2 folds at 8 h p.i.However,28 genes including FasL,CD40L,TNF,caspase-10 and caspase-3 were induced more than 2 folds after EV71 infection at 20 h.Conclusion The downregulation of apoptosis-related genes is associated with viral apoptosis-suppressing effect in RD cells in the early stage of EV71 infection.The death receptor signaling pathways including Fas/FasL and TNF/TNFR are activated to induce cell apoptosis in the late stage of EV71 infection.Moreover,host cell can also inhibit apoptosis by regulating signal pathway of CD40/CD40L,NF-κB/RelA and PI3K/Akt activation.

Objective Survey the classification on diseases of patients in hospital. Discuss personnel arrange-ment of nursing staff. Methods Survey and star sickbed number,CD rate/month,nurse number accounted on nursing level and sickbed-nurse ratio in 2007, discuss personnel arrangement of nursing staff. Results It is different that the nurse number accounted by two means, Z=2.234,P=0.025. The correlation about CD rate and nurse number in theories: r=0.782,p=0.004, nurse number in theories= CD ratex0.51-17.11, F=16.543,p=0.003.Conclusion CD rate should be reasonable personnel arrangement of nursing staff.

Parthenogenetic activation is a procedure that an oocyte at meiosis II stage is activated into mitosis by some chemical or physical stimulation other than a sperm and the embryo is formed in the absence of any contribution from a male gamete. The activation of oocyte is the result of calcium ion oscillations and deactivation of some cytokines such as maturation promoting factor, mitogen-activated protein kinase and cytostatic factor. Parthenogenetic activation is artificially induced by various kinds of physical and/or chemical methods. The main activation method of human oocyte is chemical methods. The rates of activation and cleavage depend on the age, origin,and culture conditions of the oocyte.
Adenine ; analogs & derivatives ; pharmacology ; Animals ; Calcium ; metabolism ; Cycloheximide ; pharmacology ; Cytokines ; metabolism ; Female ; Humans ; Oocytes ; drug effects ; growth & development ; metabolism ; Parthenogenesis ; drug effects

Objective:To compare the efficacy of a single injection of pegylated filgrastim with daily doses of filgrastim as pro-phylaxis for chemotherapy-induced neutropenia in Chinese cancer patients. Methods:Single-institution data from a phase 2 study and a phase 3 trial on pegylated filgrastim were combined to analyze the efficacy and safety parameters. In the two randomized crossover tri-als, patients with previously untreated cancers received two cycles of chemotherapy with identical regimen. In the study cycle, the pa-tients received a single subcutaneous injection of pegylated filgrastim (100 μg/kg), whereas those in the control cycle received daily subcutaneous injections of filgrastim (5μg/kg). Results:Among the 56 patients enrolled, 53 were evaluable for efficacy. These patients received one cycle with pegylated filgrastim prophylaxis and one cycle with filgrastim support each. Results indicated that 94.3%(50/53) of the cycles with pegylated filgrastim or filgrastim support did not develop grade 4 neutropenia. Moreover, febrile neutropenia did not occur in the cycles. The incidence rates of antibiotic administration were 7.5%(4/53) and 3.8%(2/53) in the pegylated filgrastim and filgrastim cycles, respectively (P=0.678). The median duration of filgrastim administration was 10 days (3-14 days). Generally, the safety profile of pegylated filgrastim is similar to that of filgrastim, including skeletal pain, pain at the injection site, palpitation, fever, and fatigue. Conclusion:A single dose of pegylated filgrastim demonstrated comparable efficacy with 10 consecutive doses of filgras-tim as prophylaxis for chemotherapy-induced neutropenia.

ObjectiveTo evaluate the effect of Bone mesenchymal stem cells(BMSCs) intraarticular injection on the pathological development of collagen-induced arthritis(CIA) rats.MethodsCIA Wistar rats were divided into 2 groups:the saline injection control group and the BMSCs injection group.BMSCs were isolated from normal rats and cultured in vitro till P2 generation,which were injected into the affected the ankle joint consequently.After 2,4,8,12 weeks of processing,the following indexes were observed:arthritis score and X-ray.Twelve weeks later,all of the rats were sacrificed and the following histological indexes were evaluated:cartilage destruction and cartilage extracellular matrix secretion.Paired samples t-test was used for the statistical comparison.ResultsAll the arthritis indexes of the BMSCs group were much lower.than those of the saline control group.The arthritis scores were 3.18±0.62 vs 3.84±0.35 (at week 2),3.45±0.28 vs 5.96±0.48(at week 4),3.86±0.23 vs 6.75±0.36(at week 8),4.23±0.43 vs 7.86±0.66 (at week 12) respectively.X-ray scores were 2.04±0.21 vs 2.72±0.15(at week 2),2.52±0.47 vs 4.06±0.38 (atweek 4),3.56±0.29 vs 4.35±0.36 (at week 8),3.73±0.43 vs 4.86±0.62(at weekl2) respectively and the histological evaluation indexes were 2.34±0.22 vs 3.52±0.55 (at week 12).ConclusionBMSCs injection may inhibit the pathological development of CIA rats,and it may be a promising approach for the treatment of rheumatoid arthritis.