In recent years, the advances in terahertz applications have stimulated interest in the biological effects associated with this frequency range. We study the gene expression profile in three types of cells exposed to ...In recent years, the advances in terahertz applications have stimulated interest in the biological effects associated with this frequency range. We study the gene expression profile in three types of cells exposed to terahertz radiation,i.e., human ARPE-19 retinal pigment epithelial cells, simian virus 40-transformed human corneal epithelial cells, and human MIO-M1 Müller cells. We find that the gene expression in response to heat shock is unaffected, indicating that the minimum temperature increases under controlled environment. The transcriptome sequencing survey demonstrates that 6-hour irradiation with a broadband terahertz source results in specific change in gene expression and also the biological functions that are closely related to these genes. Our results imply that the effect of terahertz radiation on gene expression can last over 15 hours and depends on the type of cell.展开更多
Acute liver failure(ALF),a fatal clinical disease featured with overwhelming hepatocyte necrosis,is a grand challenge in global health.However,a satisfactory therapeutic option for curing ALF is still absent,other tha...Acute liver failure(ALF),a fatal clinical disease featured with overwhelming hepatocyte necrosis,is a grand challenge in global health.However,a satisfactory therapeutic option for curing ALF is still absent,other than liver transplantation.Nanobiomaterials are currently being developed for the diagnosis and treatment of ALF.The liver can sequester most of nanoparticles from blood circulation,which becomes an intrinsic superiority for nanobiomaterials targeting hepatic diseases.Nanobiomaterials can enhance the bioavailability of free drugs,thereby significantly improving the therapeutic effects in ALF.Nanobiomaterials can also increase the liver accumulation of therapeutic agents and enable more effective targeting of the liver or specific liver cells.In addition,stimuli-responsive,optical,or magnetic nanomaterials exhibit great potential in the therapeutical,diagnostic,and imaging applications in ALF.Therefore,therapeutic agents in combination with nanobiomaterials increase the specificity of ALF therapy,diminish adverse systemic effects,and offer a multifunctional theranostic platform.Nanobiomaterial holds excellent significance and prospects in ALF theranostics.In this review,we summarize the therapeutic mechanisms and targeting strategies of various nanobiomaterials in ALF.We highlight recent developments of diverse nanomedicines for ALF therapy,diagnosis,and imaging.Furthermore,the challenges and future perspectives in the theranostics of ALF are also discussed.展开更多
Establishing the hepatoma cell-specific expression of human interferon gene mediated by retroviral vectors. Methods: Human interferon-β complementary DNA (IFN-β cDNA) was inserted into polylinker site of pMNSM retro...Establishing the hepatoma cell-specific expression of human interferon gene mediated by retroviral vectors. Methods: Human interferon-β complementary DNA (IFN-β cDNA) was inserted into polylinker site of pMNSM retroviral vector to construct recombinant retroviral vector pMNSIFNB, where the transcription of IFN-β gene was driven by SV40 early region promoter, and MNAIFNB, where the transcription of IFN-β gene was driven by SV40 early region promoter regulated by α-fetoprotein enhancer. The retroviral constructs were respectively introduced into PA317 amphotropic packaging cells by means of lipofectamine mediated gene transfer procedure. The plasmids transfection efficiency was among (4-25)ｘ103 colonies/μg DNA/106 PA317 cells. The retrovirus infection efficiency was among (4. 5-500)ｘ104 Colony Forming Units (CFU)/ml. The recombinant retroviruses were used to infect human hepatoma cells, renal cell carcinoma cells and melanoma cell lines in the presence of 4 μg/ml polybrene. Results: Dot hybridization of total RNA from the neomycin resistant colonies and interferon expression assay indicated that human α-fetoprotein enhancer induced efficient and specific transcription and expression of IFN-β gene driven by the promoter of different origin in human hepatoma cells by which α-fetoprotein was highly produced. Conclusion: Cis-active element of α-fetoprotein gene can drive IFN-β gene specifically expressed in human hepatoma cells, which presents some valuable materials for the hepatoma-specific immune gene therapy.展开更多
目的:探讨重组腺病毒载体在真核细胞中稳定表达阳离子抗菌肽——β-防御素2的可行性。方法:采用RT-PCR方法扩增得到大鼠β-防御素2(rBD 2)基因片段,定向插入到腺病毒穿梭质粒pShuttle-CMV中,重组质粒pShuttle-CMV-rBD 2转化E.coli B J51...目的:探讨重组腺病毒载体在真核细胞中稳定表达阳离子抗菌肽——β-防御素2的可行性。方法:采用RT-PCR方法扩增得到大鼠β-防御素2(rBD 2)基因片段,定向插入到腺病毒穿梭质粒pShuttle-CMV中,重组质粒pShuttle-CMV-rBD 2转化E.coli B J5183-AD-1后,与腺病毒基因组质粒pA dE asy-1发生同源重组,重组质粒pA dE asy-rBD 2转染293细胞进行腺病毒表达载体的包装。将包装成功的腺病毒感染cos-7细胞,并建立SD大鼠腺病毒感染模型,进行β-防御素2多肽的体外和体内表达,采用W estern b lot与免疫组化方法检测其表达。结果:重组腺病毒表达载体构建成功,包含大鼠β-防御素2基因,滴度达到109PFU/m l,W estern b lot与免疫组化方法分别检测到β-防御素2在SD大鼠体外、体内的表达。结论:重组腺病毒载体能在真核细胞中稳定表达阳离子抗菌肽——β-防御素2。展开更多
基金Project supported by the National Natural Science Foundation of China(Grant Nos.61675151 and 81570872)the Tianjin Municipal Science and Technology Commission Grants,China(Grant No.15JCYBJC24900)the Clinical Research Foundation of Tianjin Medical University Eye Institute,China(Grant No.16YKJS002)
文摘In recent years, the advances in terahertz applications have stimulated interest in the biological effects associated with this frequency range. We study the gene expression profile in three types of cells exposed to terahertz radiation,i.e., human ARPE-19 retinal pigment epithelial cells, simian virus 40-transformed human corneal epithelial cells, and human MIO-M1 Müller cells. We find that the gene expression in response to heat shock is unaffected, indicating that the minimum temperature increases under controlled environment. The transcriptome sequencing survey demonstrates that 6-hour irradiation with a broadband terahertz source results in specific change in gene expression and also the biological functions that are closely related to these genes. Our results imply that the effect of terahertz radiation on gene expression can last over 15 hours and depends on the type of cell.
基金supported by the National Key Research and Development Program of China(2019YFA0111300)the National Natural Science Foundation of China(21907113,51903256,32001012)the Guangdong Province Science and Technology Innovation Special Fund(International Scientific Cooperation,2018A050506035).
文摘Acute liver failure(ALF),a fatal clinical disease featured with overwhelming hepatocyte necrosis,is a grand challenge in global health.However,a satisfactory therapeutic option for curing ALF is still absent,other than liver transplantation.Nanobiomaterials are currently being developed for the diagnosis and treatment of ALF.The liver can sequester most of nanoparticles from blood circulation,which becomes an intrinsic superiority for nanobiomaterials targeting hepatic diseases.Nanobiomaterials can enhance the bioavailability of free drugs,thereby significantly improving the therapeutic effects in ALF.Nanobiomaterials can also increase the liver accumulation of therapeutic agents and enable more effective targeting of the liver or specific liver cells.In addition,stimuli-responsive,optical,or magnetic nanomaterials exhibit great potential in the therapeutical,diagnostic,and imaging applications in ALF.Therefore,therapeutic agents in combination with nanobiomaterials increase the specificity of ALF therapy,diminish adverse systemic effects,and offer a multifunctional theranostic platform.Nanobiomaterial holds excellent significance and prospects in ALF theranostics.In this review,we summarize the therapeutic mechanisms and targeting strategies of various nanobiomaterials in ALF.We highlight recent developments of diverse nanomedicines for ALF therapy,diagnosis,and imaging.Furthermore,the challenges and future perspectives in the theranostics of ALF are also discussed.
文摘Establishing the hepatoma cell-specific expression of human interferon gene mediated by retroviral vectors. Methods: Human interferon-β complementary DNA (IFN-β cDNA) was inserted into polylinker site of pMNSM retroviral vector to construct recombinant retroviral vector pMNSIFNB, where the transcription of IFN-β gene was driven by SV40 early region promoter, and MNAIFNB, where the transcription of IFN-β gene was driven by SV40 early region promoter regulated by α-fetoprotein enhancer. The retroviral constructs were respectively introduced into PA317 amphotropic packaging cells by means of lipofectamine mediated gene transfer procedure. The plasmids transfection efficiency was among (4-25)ｘ103 colonies/μg DNA/106 PA317 cells. The retrovirus infection efficiency was among (4. 5-500)ｘ104 Colony Forming Units (CFU)/ml. The recombinant retroviruses were used to infect human hepatoma cells, renal cell carcinoma cells and melanoma cell lines in the presence of 4 μg/ml polybrene. Results: Dot hybridization of total RNA from the neomycin resistant colonies and interferon expression assay indicated that human α-fetoprotein enhancer induced efficient and specific transcription and expression of IFN-β gene driven by the promoter of different origin in human hepatoma cells by which α-fetoprotein was highly produced. Conclusion: Cis-active element of α-fetoprotein gene can drive IFN-β gene specifically expressed in human hepatoma cells, which presents some valuable materials for the hepatoma-specific immune gene therapy.
文摘目的:探讨重组腺病毒载体在真核细胞中稳定表达阳离子抗菌肽——β-防御素2的可行性。方法:采用RT-PCR方法扩增得到大鼠β-防御素2(rBD 2)基因片段,定向插入到腺病毒穿梭质粒pShuttle-CMV中,重组质粒pShuttle-CMV-rBD 2转化E.coli B J5183-AD-1后,与腺病毒基因组质粒pA dE asy-1发生同源重组,重组质粒pA dE asy-rBD 2转染293细胞进行腺病毒表达载体的包装。将包装成功的腺病毒感染cos-7细胞,并建立SD大鼠腺病毒感染模型,进行β-防御素2多肽的体外和体内表达,采用W estern b lot与免疫组化方法检测其表达。结果:重组腺病毒表达载体构建成功,包含大鼠β-防御素2基因,滴度达到109PFU/m l,W estern b lot与免疫组化方法分别检测到β-防御素2在SD大鼠体外、体内的表达。结论:重组腺病毒载体能在真核细胞中稳定表达阳离子抗菌肽——β-防御素2。
