1.Hemophilia B replacement therapy drugs.
Chinese Journal of Biotechnology 2016;32(2):164-171
Hemophilia B is an X chromosome linked hereditary hemorrhagic disease, which is caused by the lose function mutation of factor IX (FIX), and significantly affects the patients' lifespan and life quality. The severity of hemophilia B depends on the FIX level in the plasma. By referring to the relevant literatures, we reviewed and summarized hemophilia B replacement therapies. Specifically, we focus on recombinant factor IX products on the market and those in the pipeline, especially on the long-acting factor IX drugs, to provide the basis for researches of new hemophilia B drugs.
Factor IX
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genetics
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therapeutic use
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Hemophilia B
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drug therapy
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Humans
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Mutation
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Recombinant Proteins
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therapeutic use
2.Prevention of pancreatitis after endoscopic retrograde cholangiopancreatography with different methods:a Meta analysis
Wenfei LIAO ; Hongling WANG ; Ping JIANG ; Dingyu PAN ; Fanyi ZENG
Chinese Journal of Digestive Surgery 2014;13(9):702-708
Objective To evaluate the efficacy of different methods in preventing pancreatitis after endoscopic retrograde cholangiopancreatography (ERCP).Methods Databases including PubMed,EMBASE,Cochrane Library,Chinese Journal Full-text Database,China Biomedicine Database were searched with key words including endoscopic retrograde cholangiopancreatography,ERCP,post-ERCP pancreatitis,pancreatitis,pancreatic duct stent,non-steroid anti-inflammatory drugs,indometacin,diclofenac,protease inhibitors,nafamostat,ulinastatin,gabexate,somatostain,内镜逆行胰胆管造影,内镜逆行胰胆管造影术后胰腺炎,胰腺炎,胰管支架置入,非甾体类抗炎药,吲哚美辛,双氯芬酸,抑酶剂,萘莫司他,乌司他丁,加贝酯and生长抑素.Literatures published between January 2000 and January 2014 were searched.Randomized controlled studies on prevention of pancreatitis after ERCP which were enrolled in this study were analyzed by 2 independent reviewers.The quality of the literatures was evaluated.All data were analyzed using the RevMan 5.0 software.Data were expressed in odds ratio (OR) and 95% confidence interval (95% CI).The heterogeneity of the studies was analyzed using the I2 test.Results Twenty-seven literatures were enrolled in the study.There were 4 701 patients in the experimental group (including patients who were treated by pancreatic stent installation,non-steroidal antiinflammatory drugs,nafamostat,ulinastatin,gabexate,intravenous infusion of somatostain for more than 6 hours,intravenous infusion of somatostain for less than 6 hours,bolus injection of somatostain) and 3 592 patients in the control group (including patients treated without pancreatic duct installation or placebo).The results of Meta analysis showed that pancreatic stent installation,non-steroid anti-inflammatory drugs,nafamostat,intravenous infusion of somatostain for more than 6 hours and bolus injection of somatostain could significantly decrease the incidence of pancreatitis after ERCP (OR =0.18,0.45,0.31,0.33,0.25,95% CI:0.09-0.35,0.33-0.61,0.19-0.52,0.20-0.56,0.11-0.55,P < 0.05).Conclusion Pancreatic stent installation,non-steroid anti-inflammatory drugs,nafamostat,intravenous infusion of somatostain for more than 6 hours and bolus injection of somatostain could effectively prevent the incidence of pancreatitis after ERCP.
3.An Expedient Reliable Double Fluorescent Reporter System for φC31 Integrase Function Evaluation
Huanyu XU ; Qingwen MA ; Zhaorui REN ; Zhijuan GONG ; Shuzhen HUANG ; Fanyi ZENG ; Yitao ZENG
Progress in Biochemistry and Biophysics 2009;36(7):929-933
A reporter system for φC31 integrase was developed in NIH3T3 cells. The reporter plasmid coding green fluorescent protein (GFP) coupled with red fluorescent protein (RFP) was eo-transfected with the plasmid coding φC31 integrase, to show the activity of integrase in the cells. Fluorescence activated cell sorter (FACS) was used to measure the proportion of the cells containing red and green fluorescence. The increment of green cells was positively related to the increase in the transfection with plasmid coding φC31 integrase. Approximately 90% of green cells were observed under a ratio of [plasmid-φC31-integrase]/[reporter plasmid] at 10 : 1. This suggests that the φC31 integrase reporter system provides a probe for the function of φC31 integrase in cells.
4.An Expedient Reliable Double Fluorescent Reporter System for ?C31 Integrase Function Evaluation
Huanyu XU ; Qingwen MA ; Zhaorui REN ; Zhijuan GONG ; Shuzhen HUANG ; Fanyi ZENG ; Yitao ZENG
Progress in Biochemistry and Biophysics 2006;0(07):-
A reporter system for ?C31 integrase was developed in NIH3T3 cells.The reporter plasmid coding green fluorescent protein(GFP) coupled with red fluorescent protein(RFP) was co-transfected with the plasmid coding ?C31 integrase, to show the activity of integrase in the cells.Fluorescence activated cell sorter(FACS) was used to measure the proportion of the cells containing red and green fluorescence.The increment of green cells was positively related to the increase in the transfection with plasmid coding ?C31 integrase.Approximately 90% of green cells were observed under a ratio of plasmid-?C31-integrase/reporter plasmid at 10∶1.This suggests that the ?C31 integrase reporter system provides a probe for the function of ?C31 integrase in cells.
5.Establishment and application of flow cytometry in detecting human cells in human/goat chimera models
Juan WANG ; Meijue CHEN ; Zhijuan GONG ; Zhaorui PEN ; Fanyi ZENG ; Shuzhen HUANG
Chinese Journal of Laboratory Medicine 2009;32(4):441-445
Objective To establish the methodology of flow cytometry for detecting human cells in human/goat chimerisra.Methods Human hemopoietic stem/progenitor cells (CD+34 cells) or MIG-tranadueed-GFP CD+34 cells were transplanted into the peritoneal cavity of fetal goats in utero to obtain human/goat chimera modeL The peripheral blood cells from the chimeras were labeled with multiple mouse anti-human antibodies and the monoelonal antibodies that were specific for human but had not or only minimal cross-reaction with goat were screened as the primary antibodies for routine analysis in flow cytometry.Human cord blood was proportionally (25% ,50% ,75%,100%) added into the blood of the untransplanted goats and the cells were labeled with CD+34 monoclonal antibody.The region and size of the "gate" were chosen based on to the distribution of CD+34 cells or human cord blood.One human/goat chimera marked with GFP (MIG goat) was sacrificed and the substantial liver cells from its perfused liver were analyzed for the GFP+cells percentage and DNA contents by flow cytometry.Results CD7,CD15,CD38,CD45CD20CD34CD14and GPA monoclonal antibodies were chosen as the primary antibodies in rou tine detection by flow cytometry.The size and area of the "gate" were also defined.29.1% (29100/100 000 ) of the substantial liver cells from the MIG goat expressed GFP.DNA content analysis showed that the GFP+ cells obtained from the liver of MIG goat mainly manifested two peaks that were correspond to those of human.Conclusions Flow cytometry is rapid,simple and effective for the investigation of differentiation,homing and biological characteristics of stem cells in vivo.The selections of suitable surface antibodies and the "gate" are very important for detecting human cells accurately in the human/goat chimerism.
6.Isolation and gene modification of amniotic fluid derived progenitor cells.
Chenmin YANG ; Shuyue FAN ; Huixiang TANG ; Zhijuan GONG ; Xiuli GONG ; Zhaorui REN ; Fanyi ZENG
Chinese Journal of Biotechnology 2014;30(3):492-503
We established methods to isolate human amniotic fluid-derived progenitor cells (hAFPCs), and analyze the ability of hAFPCs to secrete human coagulation factor IX (hFIX) after gene modification. The hAFPCs were manually isolated by selection for attachment to gelatin coated culture dish. hFIX cDNA was transfected into hAPFCs by using a lentiviral vector. The hFIX protein concentration and activity produced from hAFPCs were determined by enzyme-linked immunosorbent assay (ELISA) and clotting assay. The isolated spindle-shaped cells showed fibroblastoid morphology after three culture passages. The doubling time in culture was 39.05 hours. Immunocytochemistry staining of the fibroblast-like cells from amniotic fluid detected expression of stem cell markers such as SSEA4 and TRA1-60. Quantitative PCR analysis demonstrated the expression of NANOG, OCT4 and SOX2 mRNAs. Transfected hAFPCs could produce and secrete hFIX into the culture medium. The observed concentration of secreted hFIX was 20.37% +/- 2.77% two days after passage, with clotting activity of 16.42% +/- 1.78%. The amount of hFIX:Ag reached a plateau of 50.35% +/- 5.42%, with clotting activity 45.34% +/- 4.67%. In conclusion, this study established method to isolate and culture amniotic fluid progenitor cells. Transfected hAFPCs can produce hFIX at stable levels in vitro, and clotting activity increases with higher hFIX concentration. Genetically engineered hAFPC are a potential method for prenatal treatment of hemophilia B.
Amniotic Fluid
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cytology
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Blood Coagulation
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Cell Culture Techniques
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Cell Separation
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methods
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DNA, Complementary
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Factor IX
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biosynthesis
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Genetic Engineering
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Genetic Vectors
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Humans
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Stem Cells
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cytology
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metabolism
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Transfection
7.Regulation on the expression of bovine prolactin gene by different promoters.
Chinese Journal of Biotechnology 2008;24(10):1776-1782
To compare the regulation effects by different promoters on bovine prolactin gene expression in different cell lines, three recombinant bovine prolactin expression vectors were constructed using different promoters, i.e., CMV promoter, bovine prolactin gene promoter and goat beta-casein gene promoter, respectively named pCMV, pPRLP and pP1A3, which were transfected into two cell lines, mouse pituitary tumor cell strain (AtT20) and mouse mammary epithelial cell strain (HC11), respectively. RT-PCR and real-time RT-PCR were used to investigate the expression level of the above three vectors in both cell lines, pCMV vector was effectively expressed in both cell lines, pPRLP vector had a similar expression level to that of pCMV in both cell lines, pP1A3 was expressed in HC11 but not in AtT20. pP1A3 was tissue-specific to mammary gland, pPRLP was able to express with a significant level in pituitary and mammary glands, while its tissue-specific characteristics in other tissues need further investigation.
Animals
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Cattle
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Cell Line
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Cell Line, Tumor
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Epithelial Cells
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cytology
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Gene Expression Regulation
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Genetic Vectors
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Mice
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Pituitary Neoplasms
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pathology
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Prolactin
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biosynthesis
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genetics
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Promoter Regions, Genetic
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Transfection
8.Autologous peripheral blood hematopoietic stem cell transplantation in the treatment of three cases of severe pemphigus
Kang ZENG ; Lu YAN ; Jing SUN ; Fanyi MENG ; Ledong SUN ; Qifa LIU ; Dan XU ; Qian WANG ; Kuan LAI ; Zaigao ZHOU
Chinese Journal of Dermatology 2012;45(6):435-437
ObjectiveTo explore the clinical efficacy and safety of autologous peripheral blood stem cell transplantation(APBSCT) in the treatment of pemphigus.MethodsTotally,3 patients with pemphigus vulgaris who responsed poorly to 6-month treatment with glucocorticoids or immunosuppressants or experienced aggrevation of disease and developed treatment-related complications,received APBSCT and were followed up for more than 5 years.There were 1 male and 2 females with an average age of 27.3(21-39) years.The mobilization program included cyclophosphamide (CTX) 4 g/m2,recombinant human granulocyte colonystimulating factors(G-CSF) and Rituximab 375 mg/m2,and the preconditioning regimen included intravenous CTX (50 mg/kg per day on days -6,-5,-4,-3),antithymocyte globulin at 2.5 mg/kg per day(on days -3,-2,-1 and 0) and Rituximab (600 mg/d on days 0 and 7).ResultsAll the 3.patients were successfully engrafted.The mean time for peripheral reconstruction:white blood cells 13.3 days (from day 11 to 16),platelet 16.3 days (from day 16 to 17).Monitoring of immunity indices and related antibodies showed no abnormality and the immune system was well reconstructed.No serious complications occurred during the follow up,and the patients' quality of life was obviously improved.ConclusionAPBSCT may be an effective and safe option for the treatment of pemphigus.
9.Detection of autoimmune antibody in patients with SLE after autologous hematopoietic stem cell transplantation
Jianhua LI ; Jing SUN ; Xuebiao PENG ; Dan XU ; Ledong SUN ; Liang HUANG ; Fanyi MENG ; Jie ZHAO ; Kang ZENG
Journal of Chinese Physician 2002;0(S1):-
Objective To study changes of the autoimmune antibody level in patients after autologous hematopoietic stem cell transplantation(CD_(34)~+).Methods Twelve patients with SLE received autologous hematopoietic stem cell transplantation.All they survived and their immune system were recoverd after a period of time.The serum autoimmune antibody levels were measured before and after the transplantation,Results The antibody levels became normal 6 months after transplantation.Conclusion Autologous hematopoietic stem cell transplantation can effectively reduces the level of autoimmune antibody in patients with SLE.
10.Directed hepatic differentiation from embryonic stem cells.
Protein & Cell 2011;2(3):180-188
The liver is the largest internal organ in mammals, and is important for the maintenance of normal physiological functions of other tissues and organs. Hepatitis, cirrhosis, liver cancer and other chronic liver diseases are serious threats to human health, and these problems are compounded by a scarcity of liver donors for transplantation therapies. Directed differentiation of embryonic stem cells to liver cells is a promising strategy for obtaining hepatocytes that can be used for cell transplantation. In vitro hepatocyte differentiation of embryonic stem cells requires a profound understanding of normal development during embryonic hepatogenesis. Here we provide a simple description of hepatogenesis in vivo and discuss directed differentiation of embryonic stem cells into hepatocytes in vitro.
Animals
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Cell Differentiation
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Embryonic Stem Cells
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cytology
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Hepatocytes
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cytology
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metabolism
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Humans
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Liver
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cytology
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growth & development
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Signal Transduction