RESUMO
On October 5th, 2020, Drs. Harvey J. Alter, Michael Houghton and Charles M. Rice were rewarded with Nobel Prize in Physiology or Medicine for "the discovery of hepatitis C virus (HCV)". During the past 50 years, remarkable achievements have been made in treatment of HCV infection: it has changed from being a life-threatening chronic disease to being curable. In this commentary, we briefly summarized the milestone events in the "scientific journey" from the first report of non-A, non-B hepatitis and discovery of the pathogen (HCV) to final identification of efficacious direct-acting antivirals. Further, we address the challenges and unmet issues in this field.
Assuntos
Erradicação de Doenças/tendências , Hepacivirus/efeitos dos fármacos , Hepatite C/terapia , Hepacivirus/patogenicidade , Hepatite C/diagnóstico , HumanosRESUMO
AIM: To investigate human soluble PD-1 (sPD-1) biological activity sPD-1 gene be cloned and expressed in eucaryote cells. METHODS: sPD-1 gene was amplified from human PBMC mRNA by RT-PCR and cloned into eucaryotic expression vector pSG5-Fc. And the positive recombinant plasmid pSG5-Fc -sPD-1 was screened by enzyme digestion and sequencing.The correct sequencing of the recombinant plasmid pSG5-Fc-sPD-1 was transfected into COS-7 cells. The expressed recombinant protein in the supernatant was concentrated with protein A-agarose and analyzed by Western blot. The binding activity to PD-L1 which was expressed with prokaryotic cells was detected with Co-IP. RESULTS: The human sPD-1 fragment was obtained through RT-PCR. The plasmid pSG5-Fc-sPD-1 was constructed by double enzyme digestion and ligation of vector pSG5-Fc and fragment sPD-1. Sequenced sPD-1 gene was coincident with the theoretical sequence. sPD-1-Fc fusion protein in the supernatant was expressed in COS-7 cells and identified by Western blot. The activity of recombinant fusion protein sPD-1-Fc bound to PD-L1 had been detected with Co-IP. CONCLUSION: The human sPD-1 has been cloned and expressed in eucaryote cells successfully. The sPD-1-Fc fusion protein can be effective in binding to PD-L1, which can be used for further research in the function and clinical application of sPD-1.
Assuntos
Antígenos CD/metabolismo , Proteínas Reguladoras de Apoptose/metabolismo , Clonagem Molecular/métodos , Células Eucarióticas/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Animais , Antígenos CD/biossíntese , Antígenos CD/genética , Proteínas Reguladoras de Apoptose/biossíntese , Proteínas Reguladoras de Apoptose/genética , Células COS , Chlorocebus aethiops , Células Clonais , Células Eucarióticas/citologia , Vetores Genéticos , Humanos , Plasmídeos , Receptor de Morte Celular Programada 1 , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/genética , TransfecçãoRESUMO
AIM: To explore the effect of HIV-1 Nef on the ICAM-1 expression of endothelial cells ECV-304 and its signaling pathways. METHODS: ICAM-1 expression on the endothelial cells ECV-304 was detected by Western blot and FCM assay. Kinase inhibitor PD98059 was used in the cells to analyze the ERK signaling pathway. RESULTS: Western blot showed that ICAM-1 and p-ERK protein expression increased in Nef expressed ECV304 cells (ECV304-Nef) and FCM results showed that the percentage of ICAM-1 positive cells in ECV304-Nef and control cells was (35.3+/-2.2)% and (12.5+/-0.8)% respectively (P>0.01). p-ERK inhibitor PD98059 almost completely blocked the Nef up-regulation of the p-ERK and ICAM-1. When p-ERK inhibitor was added, the percentage of ICAM-1 positive cells in ECV304-Nef (11.4+/-1.1)% was reduced to the level of the control cells (10.4+/-1.5)% (P>0.05). CONCLUSION: Erk/Mapk signaling pathway may contribute to the over-expression of adhesion molecules ICAM-1 gene in HIV-1 Nef positive cells. These findings may provide the basis for further research on the mechanism and treatment of HIV-1 infection.
Assuntos
Células Endoteliais/metabolismo , Regulação da Expressão Gênica , Infecções por HIV/metabolismo , Molécula 1 de Adesão Intercelular/genética , Sistema de Sinalização das MAP Quinases , Produtos do Gene nef do Vírus da Imunodeficiência Humana/metabolismo , Células Endoteliais/enzimologia , Células Endoteliais/virologia , MAP Quinases Reguladas por Sinal Extracelular/genética , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Infecções por HIV/enzimologia , Infecções por HIV/genética , Infecções por HIV/virologia , HIV-1/genética , HIV-1/metabolismo , Humanos , Molécula 1 de Adesão Intercelular/metabolismo , Produtos do Gene nef do Vírus da Imunodeficiência Humana/genéticaRESUMO
OBJECTIVE: To investigate the effects of Chaenomeles speciosa broth on immunoregulation for anti-tumor chemotherapy. METHODS: Immunosuppressive model was induced by cyclophosphamide (CTX) in mice. The mice were treated with the broth for 15 days. The serum hemolysin was observed in mouse sera. Spleen lymphocyte transformation and gene transcription related to the immunoregulation in spleen lymphocytes were detected. RESULTS: After administrated the broth, the serum hemolysin and lymphocyte transformation rates significantly increased and the mRNA expression of foxp3, TGF-beta, PD1, Fas, Bax were downregulated compared with CTX-group. CONCLUSION: Chaenomeles speciosa broth has protective effects on the immunosuppressive mouse induce by CTX.
