Adult ; Ammonia ; poisoning ; Female ; Humans ; Microscopy, Electron ; Poisoning ; complications ; Respiratory Function Tests ; Respiratory System ; drug effects ; pathology ; ultrastructure ; Time Factors

ADP-ribosyl Cyclase 1 ; metabolism ; Aged ; Epstein-Barr Virus Infections ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Interferon Regulatory Factors ; metabolism ; Ki-67 Antigen ; metabolism ; Male ; Nasal Cavity ; Nose Neoplasms ; metabolism ; pathology ; surgery ; virology ; Plasmacytoma ; metabolism ; pathology ; surgery ; virology

AIM: To explore the clinical effects of high frequency electrical capsulotomy in maturation period cataract surgery.
●METHODS: A total of 68 cases of maturation period cataract were selected and underwent the surgery of continuous circular capsulorhexis using the high frequency electrical capsulotomy.
●RESULTS: The success rate was 91% in 68 cases with the high frequency electrical capsulotomy.
● CONCLUSION: The high frequency electrical capsulotomy in maturation period cataract surgery has significant advantages and brilliant clinical values.

Objective To obtain recombinant human Smith D1 (Sm D1) antigen and establish detecting assay.Methods Human Smith D1 antigen was synthesized by PCR using human Leukemic cDNA. The prokaryotic expression vector pGEX-ST-Sm D1 was constructed and transformed into E.coli.BL21 cell.Protein expressed under the induction of IPTG.We established DIGFA for detecting anti-Sm D1 antibodies with purified Sm D1 antigens.Results Sequence and restriction analysis revealed Sm D1 gene was cloned in frame into pGEX-5T,SDS-PAGE profile showed a clear protein band with a relative molecular weight of 39 000 and western blotting indicated that the expressed product specifically reacted to polyclonal anti-human Sm D1 genes.There was no significant difference between DIGFA and IB.The agreement between DIGFA and IB was 91.7% as calculated by Kappa statistical method.The sensitivity and specificity of DIGFA were 100% and 83.3% repectively.Conclusions Human Sm D1 gene is successfully cloned、 expressed and purification.The DIGFA,using purified Sm D1 antigens,is as good as IB,rather simpler, more rapid and reliable assay.

To clone, express and primarily use human autoantigen Sm D1 in methylotrophic yeast Pichia Pastoris. The gene Sm D1 was cloned by PCR.The PCR product was inserted into the vector pPIC9k. The recombinant plasmid pPIC9k- Sm D1 was transformed into yeast SMD1168 by electroporation. The positive clones were screened in MD plates. The high copy number transformants were rapidly selected by using G418 and were induced by methanol. Supernatants after induction were analyzed by SDS-PAGE and im-munodot. The PCR product was showed about 360 bp in size which was in accordance with predicted. The pPIC9k-Sm D1 showed the same seqencing result with GenBank’s report and restriction enzyme analysis confirmed our prediction. The pPIC9k-Sm D1 positive clone produced an about 16 kD protein which had natural immunogenicity of human autoantigen Sm D1 by SDS-PAGE and immunodot. The sensitivity and specificity of immunodot were 96% and 100%, respectively. The agreement between immunodot and im-munoblot was 98%. Successfully cloning and high-level expression of human autoantigen Sm D1 in methy-lotrophic yeast Pichia pastoris laid a foundation for further research work.

Objective To investigate the effect of MDR1 and MDR3 gene silence by shRNA of human breast carcinoma cell line MCF-7/Adr,and explore the role of MDR1 and MDR3 in adriamycin-resistance of breast carcinoma cells. Methods shRNA plasmid vector specifically targeting MDR1 and MDR3 gene was transfected into cells. The control group was transfected with empty vector. The concentration of adriamycin was detected by the flow cytometry (FCM). Cell apoptosis was analysed by FITC-Annexin-V/PI double staining. Cell viability and the IC50 of adriamycin on MCF-7/Adr cells were determined by MTT method. MDR1 and MDR3 mRNA were assessed by RT-PCR. P-gp expression was detectedby immunochemistry. Results After treatment with ABCB1 and ABCB4 shRNA plasmid vector, the apoptosis of MCF-7/Adr cells was (30.21±1.65)%and (22.07±2.17)% respectively. Compared with untransfecedgroup and empty vector transfection group the difference was significant(P＜0.01). MDR1 and MDR3 shRNAcould increase cellular adriamycin accumulation of MCF-7/Adr cells. MCF-7/Adr cells viability and the IC50were significantly decreased after transfection. Compared with untransfeced group and empty vector transfectiongroup, the mRNA level of MDR1 and MDR3 in MCF-7/Adr cells were decreased by (89.5±0.8)%and(85.1±1.2)%, the reduction of MDR1 and MDR3 mRNA was in a time-dependent manner. Immunochemistry proved that the expression of p-gp was significantly inhibited. Compared with untransfeced group and empty vector transfection group the difference was significant (P＜0.05). Conclusion The shRNA can effectively and specifically silence the expression of MDR1 and MDR3 gene, reverse the adriamycin-resistance mediated by P-gp in MCF-7/Adr cells. The reversal effect of adriamycin-resistance by shRNA of MDR1 is more effective than that of MDR3.

Adult ; Coloring Agents ; adverse effects ; Diagnosis, Differential ; Humans ; Lymphadenitis ; chemically induced ; Male ; Melanoma ; secondary ; Tattooing ; adverse effects

This paper proposes a one-way micro valve with a simple structure and a simulation design for the engineering capsule. We have now got its design parameter selection method and its mechanic characteristic from experiments.
Capsules ; Computer Simulation ; Drug Delivery Systems ; methods ; Equipment Design ; Infusion Pumps, Implantable ; Technology, Pharmaceutical ; instrumentation ; methods

OBJECTIVETo investigate the relationship between activities of early growth response gene 1 (EGR-1) of p38 mitogen-activated protein kinase (MAPK) pathway and in the epirubicin resistance of breast carcinoma cells.

METHODSProtein expression of phosphorylated p38MAPK was detected by confocal spectral microscopy. Using specific inhibitor SB203580, the effect of p38MAPK on cell apoptosis was analyzed by FITC-Annexin-V/PI double staining. The concentration of epirubicin was detected by flow cytometry (FCM). The 50% inhibition concentration (IC50) of epirubicin on MCF-7/Adr cells was determined by MTT method. Electrophoretic motility shift assay (EMSA) was performed to examine the affinity of EGR-1. EGR-1 mRNA was assessed by RT-PCR. The expression levels of p-glycoprotein, phosphorylated p53 and p38 were detected by Western blot.

RESULTSAfter treatment with SB203580 (15 micromol/L) 24 h and 48 h, (1) the early and late apoptosis of MCF-7/Adr cells expressing the phosphorylated p38MAPK protein was (25.36 +/- 1.17)% and (38.21 +/- 1.25)%, respectively, P < 0.05. And the tendency was in a time-dependent manner. (2) The average fluorescence intensity of MCF-7/Adr cells expressing the phosphorylated p38MAPK protein was (32.45 +/- 2.36) and (41.66 +/- 3.12), higher than the blank group (14.17 +/- 1.45) and DMSO group (16.28 +/- 0.63), P < 0.01. The epirubicin resistance of MCF-7/Adr cells significantly decreased. (3) SB203580 demonstrated a significantly higher level of EGR-1 activity. The IC50 was (21.53 +/- 2.17) and (8.77 +/- 1.02), lower than the DMSO group (40.74 +/- 2.56). MCF-7/Adr cells treated with SB203580 down-regulated the p38MAPK pathway activity, but up-regulated the EGR-1 mRNA expression. SB203580 significantly increased the cellular phosphorylated p53 protein level, but decreased the p-glycoprotein level in MCF-7/Adr cells.

CONCLUSIONSThere is a close relationship between p38MAPK pathway activity and the epirubicin resistance of breast carcinoma cells. The activation of EGR-1 mediated by p38MAPK pathway plays a critical role in epirubicin resistance.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Apoptosis ; drug effects ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; Early Growth Response Protein 1 ; genetics ; metabolism ; Enzyme Inhibitors ; pharmacology ; Epirubicin ; pharmacology ; Female ; Humans ; Imidazoles ; pharmacology ; Phosphorylation ; Pyridines ; pharmacology ; RNA, Messenger ; metabolism ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; metabolism

Diagnosis, Differential ; Humans ; Infant ; Male ; Neuroectodermal Tumors, Primitive ; pathology ; Orbital Neoplasms ; pathology