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کاربرد نوع شرط:
- جایگاه : پژوهشی
- مجله: Iranian Journal of Basic Medical Sciences
- نوع مقاله: Journal Article
- کلمات کلیدی: 1,Cell therapy Checkpoint protein CRISPR,Cas9 Induced tolerance PD,1 PDl
- چکیده:
- چکیده انگلیسی: Objective(s): Checkpoint blocking is considered a revolutionary method in cancer treatment. This method eliminates cancer cells by maintaining the sensitivity of immune cells. Today, cell therapy through checkpoint blocking is known as the most efficient method of cancer treatment. The programmed cell death protein-1(PD-1), as an immune check protein, has a vital role in weakening the immune responses by reducing the number of stimulated T cells. In normal situations, a decline in the immune responses can cause induced tolerance and prevent autoimmune diseases.
Materials and Methods: In this study, to reduce the induction of tolerance due to PDL-1 binding to PD-1, the PD-1 gene was destroyed in PBMCs by the means of CRISPR-Cas9 and dual-transfection of two plasmids containing the Cas 9 gene and two different sgRNAs specific to two region of PD-1 gene in order to produce a deletion mutation. Six different sgRNA were designed and cloned in PX-458 plasmid vector, and PBMCs were transfected using lipofectamine 2000 and electroporation. Indels were evaluated by gel electrophoresis and Sanger sequencing.
Results: We showed the PD-1 gene in PBMCs was knocked out successfully by CRISPR-Cas9 and dual-transfection of two sgRNAs. The minimum interval between the two sgRNAs was 448 nucleotides.
Conclusion: The results of this research demonstrated that the use of dual-transfection of CRISPR-Cas9 sgRNA is a suitable method to knock out the PD-1 gene and prevention of inducing tolerance in PBMCs.- انتشار مقاله: 18-02-1399
- نویسندگان: Seyed Ali Alambeladi,Seyed Ebrahim Hosseini,Mojtaba Jafarinia,Mehdi Dianatpour
- مشاهده
- جایگاه : پژوهشی
- مجله: Iranian Journal of Basic Medical Sciences
- نوع مقاله: Journal Article
- کلمات کلیدی: 1,Cell therapy Checkpoint protein CRISPR,Cas9 Induced tolerance PD,1 PDl
- چکیده:
- چکیده انگلیسی: Objective(s): Checkpoint blocking is considered a revolutionary method in cancer treatment. This method eliminates cancer cells by maintaining the sensitivity of immune cells. Today, cell therapy through checkpoint blocking is known as the most efficient method of cancer treatment. The programmed cell death protein-1(PD-1), as an immune check protein, has a vital role in weakening the immune responses by reducing the number of stimulated T cells. In normal situations, a decline in the immune responses can cause induced tolerance and prevent autoimmune diseases.
Materials and Methods: In this study, to reduce the induction of tolerance due to PDL-1 binding to PD-1, the PD-1 gene was destroyed in PBMCs by the means of CRISPR-Cas9 and dual-transfection of two plasmids containing the Cas 9 gene and two different sgRNAs specific to two region of PD-1 gene in order to produce a deletion mutation. Six different sgRNA were designed and cloned in PX-458 plasmid vector, and PBMCs were transfected using lipofectamine 2000 and electroporation. Indels were evaluated by gel electrophoresis and Sanger sequencing.
Results: We showed the PD-1 gene in PBMCs was knocked out successfully by CRISPR-Cas9 and dual-transfection of two sgRNAs. The minimum interval between the two sgRNAs was 448 nucleotides.
Conclusion: The results of this research demonstrated that the use of dual-transfection of CRISPR-Cas9 sgRNA is a suitable method to knock out the PD-1 gene and prevention of inducing tolerance in PBMCs.- انتشار مقاله: 18-02-1399
- نویسندگان: Seyed Ali Alambeladi,Seyed Ebrahim Hosseini,Mojtaba Jafarinia,Mehdi Dianatpour
- مشاهده
- جایگاه : پژوهشی
- مجله: Iranian Journal of Basic Medical Sciences
- نوع مقاله: Journal Article
- کلمات کلیدی: Liver,Human,Rabbit,Mesenchymal Stem Cells,Wharton jelly,Xenotransplantation
- چکیده:
- چکیده انگلیسی: Objective(s): End-stage hepatic failure is a potentially life-threatening condition for which orthotopic liver transplantation is the only effective treatment. However, a shortage of available donor organs for transplantation each year results in the death of many patients waiting for liver transplantation. Xenotransplantation, or the transplantation of cells, tissues, or organs between different species, was proposed as a possible solution to the worldwide shortage of human organs and tissues for transplantation. The purpose of this preliminary study was to reconstruct human liver tissue by xenotransplantation of human Wharton jelly mesenchymal stem cells (hWJ-MSCs) into fetal rabbit.
Materials and Methods: Isolation and confirmation of hWJ-MSCs from human umbilical cord was performed. Eight rabbits at gestational day 14 were anesthetized. All rabbits carried pregnancies to term yielding 40 rabbit fetuses. Intrauterine injection of hWJ-MSCs was performed in 24 fetuses. Twenty-seven fetuses were born alive. Ten liver samples from injected fetuses were sampled, eight rabbits 3 days after birth and two rabbits 21 days after birth. The non-injected fetuses served as positive control. Fetuses of non-injected rabbits were negative controls. Using real-time polymerase chain reaction (RT-PCR), mRNA expression of albumin (ALB), α-fetoprotein (AFP), hepatic nuclear factor 4 (HNF4), and CYP2B6 (CYP) were detected in liver samples.
Results: The human ALB, AFP, HNF4, and CYP mRNAs were expressed in the injected sampled fetuses by hWJ-MSCs into fetuses of rabbits in utero.
Conclusion: Developing xenotransplantation of hWJ-MSCs into rabbit uterus can introduce an applied approach for producing human liver tissue in rabbits.- انتشار مقاله: 02-04-1396
- نویسندگان: Leila Rezaeian,Seyed Ebrahim Hosseini,Mehdi Dianatpur,Mohammad Amin Edalatmanesh,Nader Tanideh,Asghar Mogheiseh,Amin Tamadon
- مشاهده
- جایگاه : پژوهشی
- مجله: Advanced Herbal Medicine
- نوع مقاله: Journal Article
- کلمات کلیدی: Artichoke,Insulin,Blood sugar,Liver enzymes
- چکیده:
- چکیده انگلیسی: Background and aims: Artichoke (Cynara scolymus L.) is a plant of the Asteraceae family. This plant is used in herbal medicine. This study was conducted to evaluate the effect of artichokeextract on insulin, glucagon, glucose, alkaline phosphatase (ALP), alanine amino transferase (ALT) and aspartate amino transferase (AST) in streptozotocin-induced diabetic rats. Methods: In this experimental study, 40 adult male Wistar rats weighting 220-200 g were used. The rats were divided into 4 equal groups including, control, diabetic and 2 diabetic experimental groups. Two experimental groups respectively received 100 and 300 mg/kg of artichoke hydro ethanol extract as intraperitoneal for 6 days. Diabetes was induced by intraperitoneal injection of 60 mg/kg streptozotocin. At the end of the sixth day, blood was taken from the heart of animals. Afterward serum concentration of insulin, glucagon, glucose, ALP, ALT and AST were measured. Then, the parameters measured in the different groups were compared with a control group. Data were evaluated by ANOVA, Duncan test and Student’s t-test. Results: Results showed that artichoke extract significantly reduced the levels of glucagon, glucose, ALP, ALT, AST and increased insulin level compared to control and diabetic groups. It is worth noting that before the experiment, parameters tested were equal in all groups. Conclusion: Artichoke reduces blood sugar so it has a role in treatment of diabetes. Additionally, this herb is effective in control of blood glucose by liver enzymes reducing.
- انتشار مقاله: 04-05-1393
- نویسندگان: Seyed Ebrahim Hosseini,Soghra Mousaei,Farnaz Tavakoli
- مشاهده
- جایگاه : پژوهشی
- مجله: Advanced Herbal Medicine
- نوع مقاله: Journal Article
- کلمات کلیدی: Artichoke,Insulin,Blood sugar,Liver enzymes
- چکیده:
- چکیده انگلیسی: Background and aims: Artichoke (Cynara scolymus L.) is a plant of the Asteraceae family. This plant is used in herbal medicine. This study was conducted to evaluate the effect of artichokeextract on insulin, glucagon, glucose, alkaline phosphatase (ALP), alanine amino transferase (ALT) and aspartate amino transferase (AST) in streptozotocin-induced diabetic rats. Methods: In this experimental study, 40 adult male Wistar rats weighting 220-200 g were used. The rats were divided into 4 equal groups including, control, diabetic and 2 diabetic experimental groups. Two experimental groups respectively received 100 and 300 mg/kg of artichoke hydro ethanol extract as intraperitoneal for 6 days. Diabetes was induced by intraperitoneal injection of 60 mg/kg streptozotocin. At the end of the sixth day, blood was taken from the heart of animals. Afterward serum concentration of insulin, glucagon, glucose, ALP, ALT and AST were measured. Then, the parameters measured in the different groups were compared with a control group. Data were evaluated by ANOVA, Duncan test and Student’s t-test. Results: Results showed that artichoke extract significantly reduced the levels of glucagon, glucose, ALP, ALT, AST and increased insulin level compared to control and diabetic groups. It is worth noting that before the experiment, parameters tested were equal in all groups. Conclusion: Artichoke reduces blood sugar so it has a role in treatment of diabetes. Additionally, this herb is effective in control of blood glucose by liver enzymes reducing.
- انتشار مقاله: 04-05-1393
- نویسندگان: Seyed Ebrahim Hosseini,Soghra Mousaei,Farnaz Tavakoli
- مشاهده