2.Survey on health-promoting lifestyle of kidney transplant recipients
Jiao GUO ; Xuefeng DUAN ; Hongxia FENG ; Guoxiang GUO ; Zhuo ZHANG ; Yan XIN
Chinese Journal of Organ Transplantation 2015;36(8):469-473
Objective By discussing the state of lifestyle as well as affecting factors of kidney transplant recipients,to provide more health education content and basis for their physical and psychological health.Method With internationally accepted Health-Promoting Lifestyle Profile (HPLP),we surveyed the lifestyle of the selected 104 long-term follow-up cases of kidney transplant recipients.At the same time,163 sex-and age-matched healthy volunteers,who had no obvious abnormalities in the medical test,were chosen.Result For HPLP scores among the 104 cases of recipients,20 cases were excellent,67 cases good,17 cases common,and none was inferior.As a whole,its excellent rate was 83.65%.As for 129 healthy volunteers,their overall excellent rate was 70.55%.The HPLP scores for the nutrition behavior ranked top in kidney transplant recipients,followed by healthy responsibility behavior,and lowest for exercise behavior.For the healthy volunteers,the HPLP scores for interpersonal support behavior ranked top,followed by nutrition behavior,and lowest for healthy responsibility.Correlation analysis revealed that the HPLP scores in kidney transplant recipients were significantly and positively correlated with age (r =0.307,P =0.002) and educational level (r=0.370,P =0.000),and not with gender,ethnicity,occupation and kidney sources (P>0.05).The HPLP scores in idney transplant recipients were higher than those in healthy volunteers,among which self-actualization and healthy responsibility showed statistically significant differences (P < 0.05),there was no significant difference in exercise,nutrition,interpersonal support and stress management between recipients and healthy volunteers (P>0.05).Conclusion The HPLP scores in kidney transplants was higher than in healthy volunteers,thereinto,stress management behavior and exercise behavior were relatively weak,which were the focused improvement projects of lifestyle of kidney transplant recipients.
3.A Progress on the Effects of Resveratrol on Tumor Cells
Junjun HAN ; Yuming ZHANG ; Xuefeng PAN ; Song XIE ; Fei DUAN ; Hongquan LI
World Science and Technology-Modernization of Traditional Chinese Medicine 2016;18(12):2176-2181
Resveratrol is a natural polyphenolic substance with a variety of bioactivities,such as anti-oxidation,anti-tumor and apoptosis induction.This paper summarized the recent research progress of resveratrol induced cancerous cell apoptosis,including human cervical cancer HeLa cells,leukemia cells,hepatoma cells and gastric cancer cells,and analyzed the molecular mechanism behind resveratrol-induced cell apoptosis in tumor.In addition,resveratrol was investigated by blocking the cell cycle,regulating the expressions of related genes and proteins and mitochondria-induced apoptosis pathways as a potential application of cancer drugs to clinical researches.
4.Site-specific integration and stable expression of exogenous protein at a novel site on CHO cell chromosome
Wanwan HU ; Xuefeng DING ; Yanfei CAI ; Yun CHEN ; Zuoying DUAN ; Jian JIN ; Huazhong LI
Journal of China Pharmaceutical University 2021;52(4):487-495
Finding stable expression sites on the chromosomes of Chinese hamster ovary (CHO) cells is an effective method to solve the problem of unstable expression of CHO cells in long-term culture. Our group used lentiviral transfection to integrate the tracer gene (Zsgreen1) into the chromosome of CHO cells and found multiple potential stable expression sites. This study verified the ability of one of the sites located in the 148052-148157 bp region on chromosome NW_003614241.1 to stably express exogenous proteins.The expression of Zsgreen1 gene was first observed, and CRISPR/Cas9 technology was then used to integrate the enhanced green fluorescent protein (EGFP) gene into this site. Three strains of EGFP gene integrated cells were obtained. After 60 generations of suspension culture, the fluorescence intensity of the cells had no significant changes, which proved that this site can stably express the EGFP gene. The same method was used to construct recombinant CHO cell lines expressing the human serum albumin (HSA) gene, and was verified by Western blot that this site could express and secrete HSA. It shows that the above-mentioned sites can be integrated and can stably express exogenous proteins.
5.Clinical study of Chinese herbal prescription on lipid metabolic disorders in hemodialysis patients
Guosheng ZHANG ; Mingliang DUAN ; Hongchao ZHU ; Panke ZHANG ; Xuefeng XU ; Gaiyan YUE
International Journal of Laboratory Medicine 2018;39(12):1417-1420
Objective To explore the clinical effect of traditional Chinese medicine on lipid metabolism dis-orders in hemodialysis patients.Methods 70 patients with hemodialysis received in the hospital from Novem-ber 2015 to April 2017 were enrolled in the study.They were randomly divided into control group and treat-ment group with 35 cases in each group.The control group was treated with simvastatin and and the treatment group was treated with self-made fat recipe combined with simvastatin.The levels of serum lipid ,serum in-flammatory cytokines ,renal function and liver function ,the total effective rate of treatment and the incidence of adverse reactions were compared between the two groups before and after treatment.Results There were no statistical significances in serum lipid indexes such as triglyceride (TG) ,total cholesterol (TC) ,high densi-ty lipoprotein cholesterol (HDL-C) ,low density lipoprotein cholesterol (LDL-C) ,C reactive protein (CRP) , interleukin 6 (IL-6) ,tumor necrosis factor α(TNF-α) and other serum inflammatory factors ,and renal func-tion indexes ,including urea nitrogen (BUN) ,creatinine (Scr) ,liver function indexes ,including alanine amin-otransferase (ALT) ,aspartate aminotransferase (AST) between two groups before treatment (P>0.05) ;af-ter treatment ,the TG ,TC ,LDL-C ,CRP ,IL-6 ,TNF-α ,BUN ,Scr ,ALT and AST levels of the patients in the treatment group were significantly lower than those in the control group ;the HDL-C level of the patients in the treatment group was significantly higher than that in the control group ;the total effective rate of the treat-ment group was significantly higher than that in the control group ;the incidence of adverse reactions was sig-nificantly lower than that of the control group ,and the differences were statistically significant (P>0.05). Conclusion The lipid-lowering effect of treatment of hemodialysis patients with traditional Chinese medicine lipid-lowering treatment is remarkable ,and worthy of promotion.
6.In vitro expression of human factor VIII gene induced by sodium butyrate.
Jun YIN ; Hongli WANG ; Xuefeng WANG ; Haiyan CHU ; Dao LI ; Hongbing CHEN ; Qihua FU ; Baohua DUAN ; Wenying KANG ; Qiulan DING ; Zhengwu QI ; Zhenyi WANG
Chinese Journal of Hematology 2002;23(9):463-465
OBJECTIVETo explore the effect and mechanism of sodium butyrate on expression of human clotting factor VIII in vitro.
METHODSMouse NIH/3T3 cell line was transfected with recombinant plasmid vector pRC/RSV-BDD-hFVIII, which enclosed B-domain deleted (760aa approximately 1 639aa) human factor VIII cDNA (BDD-hFVIII cDNA). Then cells were incubated in Dulbecco's modification of Eagle's medium (DMEM) containing sodium butyrate for 24 hours, hFVIII: C and hFVIII: Ag in the cell culture medium were measured by ELISA assay and one-stage method, respectively. In addition, the effect of sodium butyrate on transcription of cDNA encoding the whole hFVIII, heavy and light chain of hFVIII was also investigated by means of run-on assay.
RESULTSAfter stimulation of sodium butyrate, the levels of hFVIII: C and hFVIII: Ag increased 70% than those of control. Run-on assay showed that sodium butyrate enhanced the transcription of cDNA which encoded heavy chain of hFVIII.
CONCLUSIONSodium butyrate can improve the expression of hFVIII through enhancing the transcription of hFVIII heavy chain encoding cDNA. It demonstrated that sodium butyrate had potential utility in inducing the expression of hFVIII in vitro.
3T3 Cells ; Animals ; Butyrates ; pharmacology ; Factor VIII ; genetics ; Gene Expression Regulation ; drug effects ; Humans ; Mice
7.Study on plasma coagulation factor VII (FVII) levels and polymorphisms of FVII gene in patients with coronary heart disease.
Wenying KANG ; Hongli WANG ; Lifan XIONG ; Xuefeng WANG ; Haiyan CHU ; Bin QU ; Xiangfan LIU ; Jun YIN ; Baohua DUAN ; Jinde YU ; Zhenyi WANG
Chinese Journal of Hematology 2002;23(9):457-459
OBJECTIVETo investigate the plasma levels of coagulation factor VII (FVII) and polymorphisms of FVII gene in patients with coronary heart disease (CHD), and evaluate the effect of plasma FVII levels and FVII gene polymorphisms on CHD.
METHODSPlasma FVIIa, FVII: Ag and FVIIc were measured and polymorphisms of FVII gene were analyzed in 149 control cases and 60 CHD cases, including 33 acute myocardial infarction (AMI) cases by a combination of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and agarose gel electrophoresis.
RESULTSFVIIa, FVIIc in AMI group were significantly higher than that in control group, but FVII: Ag wasn't. There were no significant difference in plasma FVIIa, FVII: Ag and FVIIc between CHD and control group. The IVS7 genotypic frequency in AMI group was significantly different from that in control group. There was no significant difference in genotypic frequencies and allelic frequencies in other polymphism sites. FVII: Ag was significantly higher in -402A homozygote than that in -402G homozygote.
CONCLUSIONSIncreased FVII levels, especially FVIIa and FVIIc in plasma, may contribute to coronary artery thrombosis. There was significant difference in IVS7 genotype frequency between control and AMI groups, but the rest weren't significantly different. FVII: Ag was significantly higher in -402A homozygote than that in -402G homozygote. Polymorphism of -402 G/A may play an indirect role in AMI by regulating plasma FVII levels.
Coronary Disease ; blood ; genetics ; Factor VII ; analysis ; genetics ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length
8.Preliminary experimental research on gene therapy for hemophilia A.
Jun YIN ; Hongli WANG ; Yiqun HU ; Xuefeng WANG ; Bin QU ; Haiyan CHU ; Baohua DUAN ; Wenying KANG ; Zhengwu QI ; Zhenyi WANG
Chinese Journal of Hematology 2002;23(3):138-142
OBJECTIVETo accomplish a kind of therapeutic gene for hemophilia A, and observe the expression of human factor VIII (hF VIII) in vivo.
METHODSHuman clotting factor VIII cDNA with B-domain deleted (Delta760aa approximately 1639aa) was inserted into vector pRC/RSV to form pRC/RSV-hF VIII BD, which conjugated with in vivo liposome transfection reagent (DOTAP-Cholesterol) to accomplish a kind of therapeutic gene, pRC/RSV-hF VIII BD-DOTAP-Cholesterol. Mice were injected with pRC/RSV-hF VIII BD-DOTAP-Cholesterol i.m. and sacrificed 48 hours, 10 days, 20 days, 30 days, 40 days and 50 days later, respectively. Tissues such as heart, liver, spleen, lung, kidney and muscle were harvested, the distribution and transcription as well as expression of hF VIII BD cDNA were detected by means of PCR, RT-PCR and immunohistochemistry techniques. In addition, the antigen and antibody of hF VIII in plasma were measured.
RESULTSThere was high expression of hF VIII in plasma and tissues at the 48(th) hour after injection. On day 10, antigen level of hF VIII in plasma reached its peak, 17.55 ng/ml, and gradually reduced later. The antibody of hF VIII in plasma emerged on day 10 after injection, and increased and gradually reached 37.06 U/ml on day 50 after injection. PCR, RT-PCR and immunohistochemistry showed that hF VIII BD cDNA and its transcription as well as expression existed in all kinds of tissues, and lasted longer in spleen, lungs and kidneys than in heart, liver and muscle.
CONCLUSIONTherapeutic gene, pRC/RSV-hF VIII BD-DOTAP-Cholesterol, produced by combination of pRC/RSV-hF VIII BD and DOTAP-Cholesterol liposome can express human F VIII successfully in vivo, which lays an experimental foundation for curing hemophilia A by gene-drug in clinic.
Animals ; DNA, Complementary ; Disease Models, Animal ; Factor VIII ; biosynthesis ; genetics ; therapeutic use ; Gene Expression ; Genetic Therapy ; Genetic Vectors ; Hemophilia A ; therapy ; Humans ; Liposomes ; Mice ; Mice, Inbred BALB C ; Tissue Distribution ; Transfection
9.Molecular analysis of two pedigrees with hereditary F VII deficiency.
Haiyan CHU ; Hongli WANG ; Xuefeng WANG ; Xuemei GUO ; Bin QU ; Baohua DUAN ; Jun YIN ; Wenying KANG ; Zhenyi WANG
Chinese Journal of Hematology 2002;23(3):130-133
OBJECTIVETo identify the mutation of coagulation factor VII (F VII) gene in two pedigrees with hereditary F VII deficiency.
METHODSF VII gene mutations were analysed in two propositi and their family members by direct DNA sequencing. Allele specific PCR and PCR combined with restricted enzyme digestion were used to confirm the detected mutations.
RESULTSTwo gene mutations were detected in the propositus of pedigree A: G to C transition at position 6390 resulting in Trp40Cys and G to A at 11496 resulting in Arg353Gln, both are heterozygotes. The heterozygosity for polymorphism Arg353Gln was confirmed with the restriction enzyme Msp I digestion in his mother. In the propositus of pedigree B, there was a T to G transition at position 11482 resulting in His348Gln, heterozygosity of which was confirmed with Nsp I digestion in the propositus and his daughter. G to T transition at position 11514 resulting in Thr359Met was also found in the propositus of pedigree B, and the heterozygosity for Thr359Met was confirmed with allele specific PCR in the propositus and his son.
CONCLUSIONThree missense mutations were found in two pedigrees with hereditary F VII deficiency. A novel Trp40Cys mutation was reported for the first time.
Factor VII ; genetics ; Factor VII Deficiency ; genetics ; Female ; Heterozygote ; Humans ; Male ; Mutation, Missense ; Pedigree ; Point Mutation ; Polymerase Chain Reaction ; methods
10.Identification of two novel mutation in two Chinese hereditary coagulation factor XIII deficiency families.
Baohua DUAN ; Hongli WANG ; Haiyan CHU ; Xuefeng WANG ; Bin QU ; Dao LI ; Hong WANG ; Jun YIN ; Wenying KANG ; Zhenyi WANG
Chinese Journal of Hematology 2002;23(3):117-120
OBJECTIVETo explore gene defect of hereditary coagulation factor XIII deficiency.
METHODSPCR and gene sequencing or ARMS-PCR were used to detect the FXIIIA gene of peripheral white blood cell (PBC) from two Chinese hereditary coagulation factor XIII deficiency family members and 60 normal subjects respectively. The level of FXIIIA gene mRNA was tested by RT-PCR.
RESULTS(1) Nucleotide sequence analysis of the two probands' and their family members' DNA revealed that all of the three patients had homozygous missense mutation in FXIII A subunit gene. Proband 1 had a C to G transition at nucleotide (nt) 1 241 in exon 10 and proband 2 and his sister a C to T transition at nt 232 in exon 3 of FXIII A gene, which resulted in the substitution of Ser413 with Trp and Arg 77 with Cys, respectively. Family study showed that the two mutations were inherited from the parents who were correspondingly heterozygotes at nt 1 241 or nt 232. (2) The two mutations were not found in the normal subjects. (3) The FXIIIA gene mRNA level in the two probands was a little decreasing.
CONCLUSIONIt is the two novel mutations that results in FXIIIA deficiency. The two mutations of FXIIIA gene may affect its function or alter protein folding. The defective FXIII which is unstable and degraded rapidly in cytoplasm may be the main cause of FXIII deficiency.
Blood Coagulation Disorders, Inherited ; genetics ; Child ; Exons ; genetics ; Factor XIII ; genetics ; Factor XIII Deficiency ; genetics ; Female ; Heterozygote ; Humans ; Pedigree ; Point Mutation ; Polymerase Chain Reaction ; methods