ABSTRACT
Thalidomide and its derivatives have been used in the treatment of myelodysplastic syndrome (MDS) because of their anti-angiogenic and immunomodulatory effects. In recent years, some studies have found that thalidomide and its derivatives not only showed significant efficacy in lower-risk MDS patients with del (5q), but also showed advantages in non-del (5q) MDS patients. In addition, the discovery of its molecular targets and new substrates makes it possible to develop a new generation of immunomodulatory drugs (IMiDs) and to design IMiDs-based proteolysis-targeting chimeras. In this review, the new progress in mechanism and clinical application of thalidomide and its derivatives were summarized briefly, so as to provide a more scientific, reasonable and effective scheme to the treatment of MDS.
Subject(s)
Myelodysplastic Syndromes , Thalidomide , Humans , Immunomodulating Agents , Myelodysplastic Syndromes/drug therapy , Thalidomide/therapeutic useABSTRACT
AIM: To express DnaJ-homologous chaperon peripherin-binding protein(PBP) gene in E.coli and prepare the rabbit antibody against PBP. METHODS: The PBP cDNA was amplified from the human fetal brain tissue by RT-PCR. After confirmed by DNA sequencing, the PBP-cDNA was cloned into expression vector pET28a and then the PBP gene was expressed in E.coli under the IPTG induction. The expressed protein was purified through Ni-NTA affinity chromatography column. The rabbit antibody against PBP was prepared by immunizing two New Zealand white rabbits using the purified PBP as immunogen. The titer and specificity of the antisera were determined by Western blot. RESULTS: The 720 bp PBP gene was amplified, cloned, and expressed in E.coli. The expressed product existed in the bacterial inclusion body and the supernatant of the bacteria lysate. The purified PBP reached electrophoretic purity. The rabbit antibody against PBP was prepared and its titer was about 1:1,600. Western blot analysis showed that the antibody could bind to the expressed PBP protein specifically. CONCLUSION: The PBP protein was expressed in E.coli and rabbit antibody against PBP was prepared successfully, which lays the foundation for further study on the structure and biological function of PBP.
Subject(s)
Antibodies, Bacterial/immunology , Escherichia coli/genetics , HSP40 Heat-Shock Proteins/genetics , HSP40 Heat-Shock Proteins/immunology , Animals , Antibody Specificity , Cloning, Molecular , DNA, Complementary/genetics , Electrophoresis, Polyacrylamide Gel , Escherichia coli/cytology , Gene Expression , Genetic Vectors/genetics , HSP40 Heat-Shock Proteins/biosynthesis , HSP40 Heat-Shock Proteins/isolation & purification , Humans , Inclusion Bodies , RabbitsABSTRACT
AIM: To isolate and identify a human DnaJ homolog chaperon, PBP, from a human skeleton cDNA library, and to analyze its expression and distribution in transfected mammalian cells. METHODS: (32)p-dCTP labeled probe hybridization was used to screen the human skeleton cDNA library and sequence of the positive clones were analyzed. Then PBP gene was transfected into COS-7 cells using lipofectamin. PBP expressed in the cells were detected by Western-blot and indirect immunofluorescence staining. RESULTS: A full-length(1.5 kb) cDNA of peripherin-binding protein (PBP) was identified, which is identical with that of mrj. Full length PBP was mainly localized to cytoplasms of COS-7 cells in interphase, and to nuclei in mitosis. CONCLUSION: The results indicate that besides cooperating with DnaK (HSP70), PBP itself plays an important role as a member of DnaJ family. PBP may also be involved in the regulation of cell cycle.
Subject(s)
Carrier Proteins/genetics , Heat-Shock Proteins/genetics , Intermediate Filament Proteins/metabolism , Membrane Glycoproteins/metabolism , Molecular Chaperones/genetics , Nerve Tissue Proteins/metabolism , Amino Acid Sequence , Animals , CHO Cells , COS Cells , Carrier Proteins/analysis , Carrier Proteins/physiology , Cloning, Molecular , Cricetinae , HSP40 Heat-Shock Proteins , Heat-Shock Proteins/analysis , Heat-Shock Proteins/physiology , Molecular Chaperones/analysis , Molecular Sequence Data , PeripherinsABSTRACT
In order to understand insertion/delation (I/D) polymorphism of the angiotensin-converting enzyme (ACE) gene in pilots,and to explore the relationship between ACE gene I/D polymorphism and the performance of the pilots, the polymerase chain reaction (PCR) was used to determine the genotypes for an I/D polymorphism in intron 16 of the ACE gene in 118 pilots and 96 healthy subjects as controls. The result showed that the I/D polymorphism in intron 16 of the ACE gene was categorized into three genotypes: two deletion alleles (genotype DD), heterozygous alleles (genotype ID), and two insertion alleles (genotype II). The genotype II and I allele frequency were significantly higher in pilots (44.07% and 0.65) than that in healthy subjects (31.25% and 0.52). It is suggested that I gene of ACE may play a role in performance of the pilots.