Change of serum zinc-α2-glycoprotein level before and after methimazol treatment in hyperthyroidism patients with Graves disease / 重庆医学

Objective To explore the roles and possible significance of zinc-α2-glycoprotein(ZAG) in Graves disease(GD) by detecting the change of serum ZAG level before and after methimazol(MMI) treatment in the patients with initial onset of Graves disease.Methods The enzyme linked immunosorbent assay(ELISA) was adopted to detect serum ZAG level in healthy population,patients with newly diagnosed GD and GD patients treated by 12-week MMI treatment.The serum thyroid hormones and thyroid related autoantibodies were determined by adopting the electrochemiluminescence immunoassay.The blood lipid levels were examined by enzymatic colorimetry.Furthermore the relationship between serum ZAG level with thyroid function,thyroid related autoantibodies and blood lipid in the patients with initial onset of GD was analyzed.Results The levels of ZAG,FT3,FT4,TT3,TT4,TGAb,TPO and TRAb in the patients with newly diagnosed GD were much higher than those in the age-and gender-matched normal healthy population,while the TSH level was significantly lower than that in the normal control group(P＜0.05);after 12-week MMI treatment in the patients with initial onset of GD,the levels of serum ZAG,FT3,FT4,TT3,TT4 and TRAb were markedly decreased,whereas the TSH level was significantly enhanced(P＜0.05 or P＜0.01).The correlation study showed that fasting serum ZAG level in the GD patients was positively correlated with FT3,FT4,TT3 and TT4(P＜0.05);and negatively correlated with TSH,FFA,TC and TG(P＜0.05).With fasting serum ZAG in the GD patients as the dependent variable,the multiple linear regression analysis showed that TSH,TC and TG were the independent related factors of plasma ZAG level.Conclnsion The serum ZAG level is closely correlated with the occurrence and development of GD,and might play an important role in the blood lipids metabolism of GD patients.

Venous Thromboembolism Following Dantrolene Treatment for Neuroleptic Malignant Syndrome

Neuroleptic malignant syndrome (NMS) is one of the most severe iatrogenic emergencies in clinical service. The symptoms including sudden consciousness change, critical temperature elevation and electrolytes imbalance followed by mutli-organ system failure were common in NMS. In addition to aggressive interventions with intravenous fluid resuscitation and antipyretics, several antidotes have been suggested to prevent further progression of the muscle damage. Dantrolene has been reported to be one of the most effective treatments for NMS. However, the adverse effects of dantrolene treatment for NMS have not yet been evaluated thoroughly. Here we report a young male patient with bipolar I disorder who developed NMS after rapid tranquilization with haloperidol. Dantrolene was given intravenously for the treatment of NMS. However, fever accompanied with local tenderness, hardness with clear border and swelling with heat over the patient's left forearm occurred on the sixth day of dantrolene treatment. Venous thromboembolism (VTE) over intravenous indwelling site at the patient's forearm was noted and confirmed by Doppler ultrasound. The patient's VTE recovered after heparin and warfarin thrombolytic therapy. To our knowledge, this is the first case report demonstrating the possible relationship between dantrolene use and VTE in a patient with antipsychotic treatment. Although the causal relationship and the underlying pathogenesis require further studies, dantrolene should be used with caution for patients with NMS.

Screening and Identification of The Proteins Interacting With NLS-RARα Protein / 生物化学与生物物理进展

Acute promyelocytic leukemia (APL) is characterized by the generation of the prototypic promyelocytic leukemia-retinoicacid receptor alpha (PML-RARα), an oncogenic fusion protein due to chromosomal translocation. In a human myeloid cell line,PML-RARα is cleaved by neutrophil elastase (NE) to produce the mutational PML [nuclear localization signal (NLS) deletion andRARα (NLS-RARα, containing NLS of PML), both of which may play an important role in APL pathogenesis. The yeast two-hybridtechnique was used to screen the intracellular proteins interacting with NLS-RARα, which may be involved in NLS-RARα signaling. The NLS-RARα coding sequence was amplified by polymerase chain reaction method and was cloned into the bait plasmid pGBKT7vector, which, after the confirmation by sequencing, was transformed into yeast AH109 and the subsequent expression of bait plasmidwas proved by Western-blot. The transformed yeast AH109 was mated with yeast Y187 (containing leukemia cDNA library plasmidspACT2) in medium. Diploid yeast was plated on synthetic dropout nutrient medium containing X-α-gal for screening. After beingreintroduced into yeast AH109 and sequenced to verify the expression of ORF, eight positive colonies were obtained, among whichonecontaining JTV-1 was cloned. The interaction between NLS-RARα and JTV-1 was further supported by indirect immunofluorescence,GST pull-down and co-immunoprecipitation, respectively. These findings brought some new clues for the further exploration ofNLS-RARα signaling to APL.

Screening and identification of proteins interacting with RAR alpha-V via yeast two-hybrid system / 中华血液学杂志

<p><b>OBJECTIVE</b>To screen the protein interacting with retinoic acid receptor variant protein (RAR alpha-V) via the yeast two-hybrid technique (YTHT) and to find out the targets protein and study its biological function.</p><p><b>METHODS</b>The bait vector of pGBKT7-RAR alpha-V was constructed for screening proteins interacting with RAR alpha-V in K562 cell cDNA expression library via YTHT. The protein-protein interaction was confirmed with re-transformation in yeast and GST pull-down in vitro.</p><p><b>RESULTS</b>The bait vector was successfully constructed without toxicity, leakage and self-activation. Sixteen proteins were screened by YTHT and eight positive clones were identified by re-transformation in yeast. The interaction between RAR alpha-V and JTV-1 was confirmed by GST pull-down in vitro.</p><p><b>CONCLUSIONS</b>There are some kinds of proteins interacting with RAR alpha-V in cell. The biological dysfunction caused by certain protein-protein interaction may be involved in the pathogenesis of leukemia.</p>

Effects of adiponectin siRNA on the glucose transport in 3T3-L1 adipocytes / 中国病理生理杂志

AIM:To construct the adenovirus vector with adiponectin (Acrp30) siRNA, and to observe its effect on the Acrp30 expression and glucose transport in 3T3-L1 adipocytes. METHODS: Mouse Acrp30 siRNA fragment was designed, synthesized and cloned into the adenovirus vector. 3T3-L1 cells were infected with the two recombinant adenoviruses, respectively. The mRNA expression and protein levels of Acrp30 in these cells were evaluated by RT-PCR and ELISA. Glucose transport was measured by 2-Deoxy-[3H]-D-glucose incorporation method. RESULTS: The recombinant adenoviruses were successfully constructed. They remarkably downregulated the expression of Acrp30 at both mRNA and protein levels in 3T3-L1 cells, and decreased the glucose transport in 3T3-L1 adipocytes (P

Construction and identification of bait expression vector of glucocorticoid receptor binding domain / 第三军医大学学报

Objective To construct the bait expression plasmid pGBKT7-GR of glucocorticoid receptor(GR)binding domain.Methods The fragments of GR binding domain was amplified by RT-PCR,and then was cloned into pMD18-T.After being verified by sequencing,it was subcloned into the bait expression vector pGBKT7.Then the bait vector pGBKT7-GR was transformed into AH109 yeast cells and the expression of the bait protein was analyzed by Western blot.Toxicity and self-activation of the bait protein were detected.Results GR binding domain was amplified and cloned into pMD18-T and pGBKT7 successfully.The bait vector was transformed into AH109 yeast cells successfully,without toxicity or self-activation.The expression of the bait protein was confirmed by Western blot.Conclusion The successful construction of bait expression vector of glucocorticoid receptor binding domain lays the foundation for constructing small molecule ligand yeast three-hybrid system.