A Sporadic Case of Epstein Syndrome: A Rare Cause of Refractory Thrombocytopenia / 대한내과학회지

A 37-year-old female presented to our hospital with a history of bleeding episodes (excessive bleeding after tooth extraction, gum bleeding, easy bruising, and excessive menstruation) and severe thrombocytopenia (2,000/µL). She had no family history of bleeding tendency or thrombocytopenia. No peripheral lymphadenopathy or splenomegaly was noted. The patient's white blood cell count was normal; hemoglobin was 9.7 g/dL. A peripheral blood smear showed markedly decreased platelets, with occasional giant or large platelets. Bone marrow examination found increased megakaryocytes. The patient also complained of hearing difficulty; a hearing test indicated sensory-neural hearing impairment. Her thrombocytopenia was refractory to treatment with glucocorticosteroids, intravenous gamma-globulin, and danazol. In the 13 years following her initial presentation, the patient required anti-hypertensive treatment, a hearing-aid for progressive hearing loss, and started maintenance kidney dialysis. Her clinical history of refractory thrombocytopenia, progressive hearing impairment, and renal failure suggested myosin heavy chain 9 gene-related congenital syndrome (Epstein syndrome), which was confirmed by the presence of a heterozygous deletion mutation, c.221_223del, (p.Lys74del) in peripheral leukocyte deoxyribonucleic acid.

Expression and identification of recombinant fusion protein of thrombopoietin mimetic peptide and growth hormone in Pichia pastoris / 第三军医大学学报

Objective To fulfill high-efficient expression of rTMP-GH,a recombinant fusion protein of thrombopoietin mimetic peptide and human growth hormone,in Pichia pastoris.Methods cDNA fragment coding for rTMP-GH was amplified by PCR,and subsequently cloned into the expression vector pPIC9K.The linearlized plasmid pPIC9K-rTMP-GH was transformed into Pichia pastoris GS115 cells and screened in MD/MM plates under pressure of G418.The recombinant fusion protein rTMP-GH was identified by Western blotting,and its biological activity was analyzed by its ability on colony formation of megakaryoblasts.Results The pPIC9K-rTMP-GH expression plasmid was successfully constructed.GS115 cells with high expression of rTMP-GH was screened out,and the rough purified rTMP-GH showed promotive effect on megakaryoblast colony formation.Conclusion High-efficient expression of rTMP-GH in Pichia pastoris is achieved.

Effects of a new recombinant fusion protein rTMP-GH on proliferation of megakaryocytes in vitro / 第三军医大学学报

Objective To investigate the effects of rTMP-GH, a recombinant fusion protein of thrombopoietin mimetic peptide (TMP) and human growth hormone (GH), on the proliferation and thrombocytopoiesis of cultured megakaryocytes. Methods After being treated with 100 ng/ml of rTMP-GH for 7 d, Dami cells, a kind of megakaryocyte cell line, were analyzed by observing the numbers of colony forming unit. Meanwhile, Western blotting and RT-PCR were applied to detect the expression changes of globin transcription factor-1 (GATA-1), which is a main regulator of thrombocytopoiesis in megakaryocytes. Results The numbers of colony forming unit were markedly increased in cultured Dami cells incubated with rTMP-GH. Compared with normal control and GH treatment groups, both the mRNA and protein levels of GATA-1 were up-regulated significantly in Dami cells treated with rTMP-GH. Conclusion rTMP-GH has a strong ability to promote the proliferation and thrombocytopoiesis of megakaryocytes.