Objective： To evaluate the dltterentlatlon ot human umbilical cord blood ceils into hepatocyte-like cells. Methods： Mononuclear cells （MNCs） derived from human umbilical cord blood were isolated using Ficoll. The ...Objective： To evaluate the dltterentlatlon ot human umbilical cord blood ceils into hepatocyte-like cells. Methods： Mononuclear cells （MNCs） derived from human umbilical cord blood were isolated using Ficoll. The experiment was derived into 3 categories： （1） MNCs co-cultured with 50 mg minced liver tissue separated by a trans-well membrane and then collected at 0 h, 24 h, 48 h and 72 h; （2） MNCs cultured along supplemented with 100 ml/L FBS, 100 μ/ml penicillin, 100 μg/ml streptomycin, 4.7 μg/ml linoleic acid, 1×ITS, 10^-4 mol/L L-Ascorbic acid 2-P and a combination of FGF4 （100 ng/ml） and HGF （20 ng/mL）. Cells were then collected at 0 d and 16 d to examine the expression profile of hepatocyte correlating markers; （3） 0.2-0.3 ml of MNCs with a cell density of 2×10^7/ml were transplanted into prepared recipient mice [n=12, injected with 0.4 ml/kg （20%） CCl4 and 150 ng/kg 5-fluorouracil （5-Fu） prior the transplant 24 h and 48 h, respectively] via injection through tail vein. Mice were sacrificed 4 weeks after transplantation. The hepatocyte correlating mRNAs and proteins were determined by RT-PCR, immunohistochemical analysis and immunoflurence technique. Results： （1） After 72 h, a number of glycogen positive stained cells were observed with MNCs co-cultured with damaged mouse liver tissues. The expression of hepatocyte markers, human albumin （ALB）, α-fetal protein （AFP） and human GATA4 mRNA and proteins were detected by RT-PCR and immunohistochemistry as well. For the confirmation, the DNA sequencing of PCR products was performed. In control groups, MNCs co-cuhured with normal mouse hepatocytes or MNCs cultured alone, all markers remained negative. （2） In growth factor supplemented culture system, MNCs developed into larger volume with richer cytoplasm and binucleation after 16 d. Positive expression of ALB, AFP, CK18 and CK19 mRNA were detected with RT-PCR, and ALB positive staining was observed by immunocytochemistry as well. In contrast, MNCs cultured without exogenous growth factors scarcely attached to the culture dish and ALB mRNA was not detected. （3） In transplantation experiment, both of ALB and AFP mRNA were detected by RT-PCR and HSA, PCNA and ALB positive staining were observed in the livers of recipient mice by immunocytochemistry. Conclusion： MNCs from human umbilical cord blood could convert into hepatocyte-like ceils in 3 different ways, indicating their potential use in the clinic applications for the treatment of human liver diseases.展开更多
Alterations of mitochondria DNA(mtDNA)4977 bp common deletion(CD)and mtDNA copy number induced by ionizing radiation were observed in human different cell lines and total body irradiation patients.However,only few exp...Alterations of mitochondria DNA(mtDNA)4977 bp common deletion(CD)and mtDNA copy number induced by ionizing radiation were observed in human different cell lines and total body irradiation patients.However,only few experiments have evaluated the levels of the CD and mtDNA copy number in human peripheral blood exposed to ionizing radiation till now.The aim of this study is to analyze the mtDNA alterations in irradiated human peripheral blood from healthy donors as well as to explore their feasibility as biomarkers for constructing new biodosimeter.Peripheral blood samples were collected from six healthy donors,and exposed to 60Co gamma ray with the doses of 0 Gy,1 Gy,2 Gy,3 Gy,4 Gy and 5 Gy.Levels of the CD and mtDNA copy number in irradiated samples after 2h or 24h incubation were detected using TaqMan real-time PCR,and the CD ratio was calculated.The results showed that the mean of the CD ratio and the CD copy number exhibited a dose-dependent increase 2 h in the dose range from 0–5 Gy,and of the mtDNA copy number significantly increased 24 h in irradiated groups compared with 0 Gy group after irradiation.It indicates that the parameters in human peripheral blood may be considered as molecular biomarkers to applying construction of new biodosimeter.展开更多
The biomodulative and hematopoietic potentialities of IL-2 and IL-3 activatedbone marrow(ABM)cells from patients with lung adenocarcinoma were studied in vitro.Human bone marrow(BM)cells could be activated by IL-2 in ...The biomodulative and hematopoietic potentialities of IL-2 and IL-3 activatedbone marrow(ABM)cells from patients with lung adenocarcinoma were studied in vitro.Human bone marrow(BM)cells could be activated by IL-2 in culture for 7d.TheseIL-2 ABM cells had higher cytolytic activities against cells of H 7402 cell line and freshautologous adenocarcinoma cells and maintained the cytotoxicities longer than IL-2 acti-vated peripheral blood lymphocytes(APBLs),a point of possible importance in adoptiveimmunotherapy for cancer patients.The IL-2 ABM cells also had similar number ofBFU-E and CFU-GM to that had fresh BM cells if 1L-3 was added 48h alter IL-2 inculture.The IL-2 and IL-3 ABM cells might be used to eliminate tumor cells and tosupply reconstitutive elements of BM for autologous bone marrow transplantation.展开更多
Incorporation of synthetic heme(FeP) into recombinant human serum albumin(rHSA) provides an artificial hemoprotein(rHSA-FeP) which can bind and release oxygen reversibly under physiological conditions(in aqueous media...Incorporation of synthetic heme(FeP) into recombinant human serum albumin(rHSA) provides an artificial hemoprotein(rHSA-FeP) which can bind and release oxygen reversibly under physiological conditions(in aqueous media, pH 7.3, 37 ℃) like hemoglobin(Hb) and myoglobin. An rHSA host absorbs maximally eight FeP molecules, and the solution properties are almost identical to those of rHSA itself. The second-order structure and surface charge distribution of rHSA were always constant independent of the binding numbers of FeP. Its O 2-binding ability satisfies the initial clinical requirements for red cell substitute. Although the NO-binding affinity is 8-fold high compared to the Hbs, administration of this fluid into rats showed negligible change in the blood pressure. Physiological responses to exchange transfusion with this rHSA-FeP into anaesthetized rats have also been evaluated.展开更多
Cryopreservation of red blood cells(RBCs)provides great potential benefits for providing transfusion timely in emergencies.High concentrations of glycerol(20%or 40%)are used for RBC cryopreservation in current clinica...Cryopreservation of red blood cells(RBCs)provides great potential benefits for providing transfusion timely in emergencies.High concentrations of glycerol(20%or 40%)are used for RBC cryopreservation in current clinical practice,which results in cytotoxicity and osmotic injuries that must be carefully controlled.However,existing studies on the low-glycerol cryopreservation of RBCs still suffer from the bottleneck of low hematocrit levels,which require relatively large storage space and an extra concentration process before transfusion,making it inconvenient(time-consuming,and also may cause injury and sample lose)for clinical applications.To this end,we develop a novel method for the glycerol-free cryopreservation of human RBCs with a high final hematocrit by using trehalose as the sole cryoprotectant to dehydrate RBCs and using core–shell alginate hydrogel microfibers to enhance heat transfer during cryopreservation.Different from previous studies,we achieve the cryopreservation of human RBCs at high hematocrit(>40%)with high recovery(up to 95%).Additionally,the washed RBCs post-cryopreserved are proved to maintain their morphology,mechanics,and functional properties.This may provide a nontoxic,high-efficiency,and glycerol-free approach for RBC cryopreservation,along with potential clinical transfusion benefits.展开更多
基金Supported by the Shenzhen Science & Technology Planning Program (No. 200204109, No. JH200505270412B)
文摘Objective： To evaluate the dltterentlatlon ot human umbilical cord blood ceils into hepatocyte-like cells. Methods： Mononuclear cells （MNCs） derived from human umbilical cord blood were isolated using Ficoll. The experiment was derived into 3 categories： （1） MNCs co-cultured with 50 mg minced liver tissue separated by a trans-well membrane and then collected at 0 h, 24 h, 48 h and 72 h; （2） MNCs cultured along supplemented with 100 ml/L FBS, 100 μ/ml penicillin, 100 μg/ml streptomycin, 4.7 μg/ml linoleic acid, 1×ITS, 10^-4 mol/L L-Ascorbic acid 2-P and a combination of FGF4 （100 ng/ml） and HGF （20 ng/mL）. Cells were then collected at 0 d and 16 d to examine the expression profile of hepatocyte correlating markers; （3） 0.2-0.3 ml of MNCs with a cell density of 2×10^7/ml were transplanted into prepared recipient mice [n=12, injected with 0.4 ml/kg （20%） CCl4 and 150 ng/kg 5-fluorouracil （5-Fu） prior the transplant 24 h and 48 h, respectively] via injection through tail vein. Mice were sacrificed 4 weeks after transplantation. The hepatocyte correlating mRNAs and proteins were determined by RT-PCR, immunohistochemical analysis and immunoflurence technique. Results： （1） After 72 h, a number of glycogen positive stained cells were observed with MNCs co-cultured with damaged mouse liver tissues. The expression of hepatocyte markers, human albumin （ALB）, α-fetal protein （AFP） and human GATA4 mRNA and proteins were detected by RT-PCR and immunohistochemistry as well. For the confirmation, the DNA sequencing of PCR products was performed. In control groups, MNCs co-cuhured with normal mouse hepatocytes or MNCs cultured alone, all markers remained negative. （2） In growth factor supplemented culture system, MNCs developed into larger volume with richer cytoplasm and binucleation after 16 d. Positive expression of ALB, AFP, CK18 and CK19 mRNA were detected with RT-PCR, and ALB positive staining was observed by immunocytochemistry as well. In contrast, MNCs cultured without exogenous growth factors scarcely attached to the culture dish and ALB mRNA was not detected. （3） In transplantation experiment, both of ALB and AFP mRNA were detected by RT-PCR and HSA, PCNA and ALB positive staining were observed in the livers of recipient mice by immunocytochemistry. Conclusion： MNCs from human umbilical cord blood could convert into hepatocyte-like ceils in 3 different ways, indicating their potential use in the clinic applications for the treatment of human liver diseases.
基金Supported by grants from the building project for the National Key Clinical Special Department of China(No.2011-17)the Medical Science and Technology Foundation of Henan Province(No.201204123)the Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘Alterations of mitochondria DNA(mtDNA)4977 bp common deletion(CD)and mtDNA copy number induced by ionizing radiation were observed in human different cell lines and total body irradiation patients.However,only few experiments have evaluated the levels of the CD and mtDNA copy number in human peripheral blood exposed to ionizing radiation till now.The aim of this study is to analyze the mtDNA alterations in irradiated human peripheral blood from healthy donors as well as to explore their feasibility as biomarkers for constructing new biodosimeter.Peripheral blood samples were collected from six healthy donors,and exposed to 60Co gamma ray with the doses of 0 Gy,1 Gy,2 Gy,3 Gy,4 Gy and 5 Gy.Levels of the CD and mtDNA copy number in irradiated samples after 2h or 24h incubation were detected using TaqMan real-time PCR,and the CD ratio was calculated.The results showed that the mean of the CD ratio and the CD copy number exhibited a dose-dependent increase 2 h in the dose range from 0–5 Gy,and of the mtDNA copy number significantly increased 24 h in irradiated groups compared with 0 Gy group after irradiation.It indicates that the parameters in human peripheral blood may be considered as molecular biomarkers to applying construction of new biodosimeter.
文摘The biomodulative and hematopoietic potentialities of IL-2 and IL-3 activatedbone marrow(ABM)cells from patients with lung adenocarcinoma were studied in vitro.Human bone marrow(BM)cells could be activated by IL-2 in culture for 7d.TheseIL-2 ABM cells had higher cytolytic activities against cells of H 7402 cell line and freshautologous adenocarcinoma cells and maintained the cytotoxicities longer than IL-2 acti-vated peripheral blood lymphocytes(APBLs),a point of possible importance in adoptiveimmunotherapy for cancer patients.The IL-2 ABM cells also had similar number ofBFU-E and CFU-GM to that had fresh BM cells if 1L-3 was added 48h alter IL-2 inculture.The IL-2 and IL-3 ABM cells might be used to eliminate tumor cells and tosupply reconstitutive elements of BM for autologous bone marrow transplantation.
文摘Incorporation of synthetic heme(FeP) into recombinant human serum albumin(rHSA) provides an artificial hemoprotein(rHSA-FeP) which can bind and release oxygen reversibly under physiological conditions(in aqueous media, pH 7.3, 37 ℃) like hemoglobin(Hb) and myoglobin. An rHSA host absorbs maximally eight FeP molecules, and the solution properties are almost identical to those of rHSA itself. The second-order structure and surface charge distribution of rHSA were always constant independent of the binding numbers of FeP. Its O 2-binding ability satisfies the initial clinical requirements for red cell substitute. Although the NO-binding affinity is 8-fold high compared to the Hbs, administration of this fluid into rats showed negligible change in the blood pressure. Physiological responses to exchange transfusion with this rHSA-FeP into anaesthetized rats have also been evaluated.
基金the National Natural Science Foundation of China(No.82172114)the Anhui Provincial Natural Science Foundation for Distinguished Young Scholars(No.2108085J37).
文摘Cryopreservation of red blood cells(RBCs)provides great potential benefits for providing transfusion timely in emergencies.High concentrations of glycerol(20%or 40%)are used for RBC cryopreservation in current clinical practice,which results in cytotoxicity and osmotic injuries that must be carefully controlled.However,existing studies on the low-glycerol cryopreservation of RBCs still suffer from the bottleneck of low hematocrit levels,which require relatively large storage space and an extra concentration process before transfusion,making it inconvenient(time-consuming,and also may cause injury and sample lose)for clinical applications.To this end,we develop a novel method for the glycerol-free cryopreservation of human RBCs with a high final hematocrit by using trehalose as the sole cryoprotectant to dehydrate RBCs and using core–shell alginate hydrogel microfibers to enhance heat transfer during cryopreservation.Different from previous studies,we achieve the cryopreservation of human RBCs at high hematocrit(>40%)with high recovery(up to 95%).Additionally,the washed RBCs post-cryopreserved are proved to maintain their morphology,mechanics,and functional properties.This may provide a nontoxic,high-efficiency,and glycerol-free approach for RBC cryopreservation,along with potential clinical transfusion benefits.
