1.Complement mediated killing of human glomerular mesangial cell by Fcα/μ receptor
Lian SHEN ; Xiaoyan WANG ; Qing ZHAO ; Wei ZHANG
Basic & Clinical Medicine 2010;30(5):471-475
Objective To study whether Fcα/μ receptor(Fcα/μR)can mediate complement killing of human glomerular mesangial cells.Methods Fcα/μR cDNA contained plasmid,pcDNA3.1-Fcα/μR was transfected into a human glomerular mesangial cell(NHMC).Fcα/μR expression was detected by Western blot and laser scanning eonfocal microscopy.Binding of IgM-immune complexes(IgM-IC)to the Fcα/μR on cell membrane was detected by flowcytometry and laser scanning confocal microscopy.Killing of cells by complement was shown by Trypan blue exclusion assay.Results NHMC cells transfected with Fcα/μR could bind IgM and IgM-IC.After treatment with complement,added IgM-IC,the death rate of pcDNA3.1-Fcα/μR transfected cell was significant higher than the control groups of wild type cell,pcDNA3.1 transfected cell and the pcDNA3.1-Fcα/μR transfected cell without IgM-IC.Conclusion IgM-IC can bind to the Fcα/μR expressed NHMC cells and mediate complement killing of the cells.
2.Changes of body composition in children with acute leukemia during different treatment stages
Yi FENG ; Xiaoyan YU ; Jingchao LIAN ; Li HONG
Chinese Journal of Clinical Nutrition 2011;19(2):88-92
Objective To investigate the changes of body composition in the children with acute leukemia during different treatment stages.Methods From January 2009 to April 2010,56 children with acute leukemia hospitalized in Shanghai Children's Medical Center for chemotherapy were enrolled.Meanwhile,56 healthy children with matched age and sex were enrolled as the control group.The body compositions of children in the control group and the children with acute leukemia at the end of the first course of each treatment stages ware detected by segmental multiple-frequency bioelectrical impedance analysis.The distribution of body compositions was recorded asthe percentage of each body composition to the body weighL Results Among 56 children with acute leukemia,41were with acute lymphoblastic leukemia and 15 with acute nonlymphoblasdc leukemia.Twenty-three cases were in remission-induction chemotherapy stage,15 in consolidation chemotherapy stage,and 18 in maintenance chemotherapy stage.Compared with children in the control group,children with acute leukemia showed a reduction in the percentage of intracellular fluid (P = 0.000), extracellular fluid (P - 0.005), protein (P = 0.000), mineral (P = 0.001), skeletal muscle mass (P = 0.000), body cell mass (P = 0.000), and active cell mass (P = 0.000), while an increase in body fat mass [(26.2 ± 8.3)% vs.(20.3±3.8)%, P=0.000].The body weight (P = 0.001), the percentage of intracellular fluid (P = 0.005), protein (P = 0.004), body cell mass (P = 0.001), and active cell mass ( P = 0.020 ) in the children during remission-induction chemotherapy stage were significantly lower than those of the healthy children.However, the parameters of the consolidation chemotherapy stage were significantly lower than those of the control group, including the percentage of intracellular fluid (P = 0.000), extracellular fluid (P =0.000), protein (P = 0.000), mineral (P =0.001), body fat mass (P =0.000), non-fat mass (P = 0.000), skeletal muscle mass (P = 0.000), body cell mass (P = 0.000), and active cell mass (P = 0.000).Most body compositions in the maintenance chemotherapy stage were lower than those of the healthy children ( P < 0.05) except for body mass index (P = 0.127) and the percentage of extracellular fluid (P = 0.097).Conclusions Body compositions remarkably change in children with acute leukemia after chemotherapy.Therefore, the nutritional status of children with acute leukemia should be closely monitored, and proper nutritional support should be provided when necessary to improve the prognosis.
3.Endothelial nitric oxide synthase traffic inducer in the umbilical vessels of the patients with pre-eclampsia.
Wenpei, XIANG ; Hanping, CHEN ; Lian, HU ; Xiaoyan, XU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(2):243-5
The expression of endothelial nitric oxide synthase traffic inducer (NOSTRIN) was examined in the umbilical vessels of the patients with pre-eclampsia (PE) to explore its possible role in the pathogenesis of PE. The NOSTRIN mRNA in umbilical tissues was determined by RT-PCR. The eNOS activity in umbilical vessels was spectrophotometrically detected. NO2-/NO3-, the stable metabolic end products of NO, was measured by using nitrate reductase. RT-PCR showed that the expression level of NOSTRIN was significantly higher in women with PE than in the normal group (P<0.01). The activity of eNOS was significantly decreased in PE group [(12.83+/-3.61) U/mg] than in normal group [(21.72+/-3.83) U/mg] (P<0.01). The level of NO2-/NO3- in PE patients (27.53+/-7.48) micromol/mg was significantly lower than that of normal group (54.27+/-9.53) micromol/mg (P<0.01). The significant negative correlation existed between the expression of NOSTRIN and the activity of eNOS in umbilical vessels of women with PE (r=-0.58, P<0.01). It was concluded that the level of NOSTRIN expression was increased in umbilical vessel of women with PE, indicating that it may be involved in the pathogenesis of PE.
Intracellular Signaling Peptides and Proteins/genetics
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Intracellular Signaling Peptides and Proteins/*metabolism
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Pre-Eclampsia/*enzymology
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Pre-Eclampsia/etiology
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RNA, Messenger/genetics
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RNA, Messenger/metabolism
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Umbilical Arteries/cytology
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Umbilical Arteries/*enzymology
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Umbilical Veins/cytology
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Umbilical Veins/*enzymology
4.Dual-Sensitive Probe of 2-( Benzoacridin) ethyl-imidazole-1-carboxylate for Determination of Aliphatic Amine with Fluorescence and Online Atmospheric Pressure Chemical Ionization Mass Spectrometry Identification
Yanyan FU ; Xiaoyan LI ; Zhiwei SUN ; Xueqin QIN ; Lian XIA ; Yourui SUO ; Yulin LI ; Jinmao YOU
Chinese Journal of Analytical Chemistry 2010;38(1):8-14
With comparison of three different methods for the marking of amines compound, an optimal deri vatization method was selected.5-(2-Hydroxyethyl) benzoacridine (HBA) reacts with coupling agent N,N'-carbonyldiimidazole(CDI) to form an activated amide intermediate 2-(benzoacridin) ethyl-imidazole-1-carbox-ylate(BAEIC).BAEIC, which is dual-sensitive probe, reacts preferably with amino compounds at 80 ℃ in the presence of 4-dimethylaminopyridine(DMAP) catalyst in N,N-dimethylformamide(DMF) solvent to give the corresponding sensitively fluorescent derivatives with an excitation maximum at λ_(ex) of 280 nm and an emis sion maximum at λ_(em) of 510 nm.BAEIC-amine derivatives simultaneously exhibited high ionization potential with percent ionization (changing from 5.62% to 58.08% in aqueous acetonitrile and from 2.14% to 56.58% in aqueous methanol.Derivatives were not only sensitive to fluorescence but also to MS ionizable potential.The fluorescence detection limits(5/iV = 3) were 0.12-0.59 μg/L.The online APCI-MS detection limits were 1.9-14 μg/L(S/N=5).
5.Sex hormone-binding globulin of gestational diabetes mellitus pregnant women with well-controlled glucose and pregnancy outcomes
Zhen JIN ; Xinshu CHI ; Weiping TENG ; Xiaoyan WANG ; Qiwei XU ; Pingting WANG ; Wanting AI ; Siyu LIAN
Chinese Journal of Obstetrics and Gynecology 2011;46(6):422-426
Objective To explore the relationship between sex hormone-binding globulin (SHBG) of gestational diabetes mellitus ( GDM ) pregnant women with well-controlled glucose and pregnancy outcomes. Methods Two hundred and fifty-one GDM pregnant women of 24 - 28 weeks in Shengjing Hospital of China Medical University were recruited from Mar. 2005 to Mar. 2010. Two hundred and sixteen cases of GDM with well-controlled glucose were defined as glycemic satisfied group, and they were treated by diet therapy ( 169 cases) or insulin therapy (47 cases) . Thirty-five cases with unsatisfied glucose were defined as glycemic unsatisfied group. One hundred and ninety-two healthy pregnant women of 24 - 28 weeks were defined as healthy control group. Serum SHBG and homeostasis model analysis of insulin resistance ( HOMA-IR) at 24 - 28 weeks and above 36 weeks were measured. GDM was diagnosed bytwo-step method according to the National Diabetes Data Group ( NDDG) criteria. The pregnancy outcomes and complications of the three groups were recorded. Results ( 1 ) Comparison of pregnancy outcomes and complications: glycemic satisfied group was less likely to develop hypertensive disorders in pregnancy ( 10. 6% ) , premature birth(8. 3% ) ,large for gestational age ( LGA) (8. 8% ) , neonatal asphyxia(3. 7% ) and neonatal hypoglycemia ( 2. 3% ) compared to glycemic unsatisfied group ( 42. 9% , 34. 3% , 31. 4% , 22. 9% and 11. 4% ,respectively). And the difference was statistically significant (P <0. 05 or P <0. 01). There was no significant difference for incidence of polyhydramnios, pueperal infection, postpartum hemorrhage, neonatal hyperbilirubinemia between the two groups ( P> 0. 05 ) . When compared to healthy control group(7. 3% ,2. 1% ,4. 2% ,2. 1% and 1. 6% ) ,no significant difference was found for incidence of premature birth( 8. 3% ) , pueperal infection ( 3. 2% ) , postpartum hemorrhage (5. 1% ) , neonatal asphyxia (3. 7% )and neonatal hypoglycemia(2. 3% ,P >0. 05). (2) Comparison of results of 24 - 28 weeks and above 36 weeks: serum SHBG of glycemic satisfied group [( 384 ± 88 ) , (457 ± 48 ) nmol/L]was significantly higher than that of glycemic unsatisfied group[(313 ±45) ,(401 ±73) nmol/L];HOMA-IR of glycemic satisfied group (5. 3 ±1.1,5.5 ±1.1) was significantly lower than that of glycemic unsatisfied group (7. 0 ± 1. 3 ,7. 6 ± 1. 7 ; P < 0. 01). Serum SHBG of glycemic satisfied group was significantly lower than that of healthy control group [( 492 ± 95 ) , (565 ± 40 ) nmol/L]; and HOMA-IR of glycemic satisfied group(5. 3 ± 1. 1,5. 5 ± 1. 1) was significantly higher than that of healthy control group (3. 6 ±0. 6,3. 9 ± 0. 5 ;P < 0. 01 ) . FPG of glycemic satisfied group [( 5. 84 ± 0. 28 ) , ( 5. 16 ± 0. 13 ) mmol/L]was significantly lower than that of glycemic unsatisfied group [(6. 13 ± 0. 16 ) , ( 5. 68 ± 1. 14) mmol/L; P < 0. 01]. FINS of glycemic satisfied group [( 20. 4 ± 2. 1 ) , ( 24. 1 ± 4. 2 ) mmol/L]was significantly lower than that of glycemic unsatisfied group [(24. 7 ± 4. 5 ) , ( 29. 9 ± 2. 7 ) mmol/L; P < 0. 01]. ( 3 ) Correlation analysis. Between 24 - 28 weeks, SHBG was negatively correlated with HOMA-IR in the three groups ( r = -0. 952, P <0. 01) ; and SHBG was negatively correlated with HOMA-IR in glycemic satisfied group ( r = -0. 903, P <0. 01). Conclusions Well-controlled glucose can not completely improve maternal and fetal outcomes of GDM pregnant women. High insulin resistance and low serum SHBG can influence pregnancy outcomes.
6.Genotypic diagnosis of long QT syndrome by analysis of candidate genes
Jiangfang LIAN ; Chen HUANG ; Xiaoyan HUANG ; Ying WANG ; Shijun GE ; Jianqing ZHOU
Journal of Pharmaceutical Analysis 2009;21(4):222-224,229
Objective To diagnose 6 LQTS families by genetic analysis. Methods A total aof 6 LQTS pedigrees with 43 family members were brought together for genetic diagnosis by using short-sequence tandem-repeat (SIR) markers or sequencing. Genomic DNA was extracted from blood samples by standard procedure. STR markers or KCNQ1, KCNH2 and SCN5A were amplified. The haplotype analysis for LQTS was performed. If the family got the negative haplotype analysis, the sequencing was performed. Results LQTS patients were always linkaged with the SCNSA gene in family 1. KCNH2 was linkaged with the disease in family 2 to 5.21 gene carriers were identified from these 5 families. A mutation (A561V-KCNH2) was only found in the proband of family 6 and an SNP (G1691A) was found in all the members of the family. Conclusion Genetic diagnosis can not only improve presymptomatic diagnosis,bnt also provide the basis for personal therapy and research on disease-causing mutations.
7.A novel mutation-L539fs/47 of hERG in a Chinese long QT syndrome family
Jiangfang LIAN ; Xiaoyan HUANG ; Weifeng XU ; Xi YANG ; Ying WANG ; Di LI ; Jianqing ZHOU
Journal of Pharmaceutical Analysis 2010;22(3):188-191
Objective To identify the mutation of human ether-a-go-go-related gene (hERG) and analyze the clinical characteristics of a Chinese family with long ST syndrome (LQTS). Methods The electrocardiogram and DNA samples were obtained from a Chinese LQTS family of 26 members. Genotype was performed with polymorphic short tandem repeat (STR) markers at the known LQT1, LQT2, and LQT3 loci. SSCP analysis was used to find aberrant conformers. hERG mutation was confirmed by cloning and sequencing. Results Three gene carriers were linked to chromosome 7q35-36, where the potassium channel gene hERG was encoded. A 19-base pair deletion was identified. The mutation was located at nucleotide position 1 619-1 637 between transmembrane domains S4 and S5. Furthermore, A1692G polymorphism was found both in the normal control and patients. Conclusion A novel 19 bp deletion mutation of hERG is identified in a Chinese family. All gene carriers are demonstrated to be typical LQT2 ECG phenotype.
8.Application of secretary luciferase labeled orthotopic transplant model of hepatocellular carcinoma to evaluate tumor response to interferon-beta gene therapy.
Gang WANG ; Zhonghui LIAN ; Wenhong TIAN ; Xiaoyan DONG ; Jie YUCHI ; Xiaobing WU
Chinese Journal of Biotechnology 2012;28(10):1236-1244
To establish an orthotopic transplant mouse model of hepatocellular carcinoma (HCC) labeled with secretary luciferase and to study its response to anti-tumor treatment with interferon-beta gene therapy. We labeled the murine hepatoma Hepal-6 cells with secretary Gaussia princeps luciferase (Gluc), and then injected Gluc labeled Hepal-6 cells intrasplenically in C57BL/6 mice. We monitored blood Glue to evaluate the tumor development and anti-tumor effects of hydrodynamic injection with interferon-beta expressing plasmid. We successfully established the orthotopic mouse model of HCC by intrasplenic injection of Glue labeled Hepal-6 cells. The Glue blood assay could reflect the amount of cancer cells in vivo, tumor progression, as well as anti-tumor effect of interferon-beta gene therapy. In conclusion, Gluc labeled orthotopic transplant mouse model of HCC can ex vivo real-time monitor the tumor development and tumor response to treatments.
Animals
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Carcinoma, Hepatocellular
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pathology
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therapy
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Cell Line, Tumor
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Genes, Reporter
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Genetic Therapy
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Interferon-beta
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genetics
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therapeutic use
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Liver Neoplasms
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pathology
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therapy
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Luciferases
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blood
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genetics
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secretion
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Mice
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Mice, Inbred C57BL
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Neoplasm Transplantation
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Treatment Outcome
9.A novel mutation of the KCNH2 gene in a family with congenital long QT syndrome.
Jiangfang LIAN ; Jianqing ZHOU ; Xiaoyan HUANG ; Ying WANG ; Xi YANG ; Di LI
Chinese Journal of Medical Genetics 2010;27(1):77-80
OBJECTIVETo perform mutation analysis in a family with long QT syndrome.
METHODSThe medical record of the affected child and his parents were collected. The locus of gene associated with the long QT syndrome was mapped by linkage analysis. Mutation analysis was done by PCR-single strand conformation polymorphism (SSCP) and direct sequencing.
RESULTSA mutation (L539fs/47) and a SNP (L564L) were found in exon 7 of the KCNH2 gene of the proband. The mutation was from the father.
CONCLUSIONA novel mutation of L539fs/47 in the KCNH2 gene was identified in the LQTS family, which might be the disease-causing mutation for the family.
Base Sequence ; ERG1 Potassium Channel ; Ether-A-Go-Go Potassium Channels ; genetics ; Female ; Frameshift Mutation ; Humans ; Long QT Syndrome ; congenital ; genetics ; Male ; Molecular Sequence Data ; Pedigree ; Polymorphism, Single Nucleotide ; Young Adult
10.Ten years of follow-up study on committed suicide and attempted suicide in Jinuo nationality
Xiaoyan LIU ; Xiaobin YANG ; Qiuyuan LI ; Zuxin ZHU ; Hongyi LI ; Lian LIU ; Hongchun GENG ; Liqin XIE ; Peikai LI ; Yong ZENG ; Jianzhong YANG
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(4):339-341
Objective To investigate the recent decade (1999 to 2009) incidence of committed suicide and attempted suicide of the Jinuo nationality in Jinuo Mountain and study the clinical characteristics of it. Methods 10 610 subjects at 44 Jinuo villagers committees were investigated by the method of door-to-door inquiry under the help of clues and collected the data of larithmics and medical history. Results 32 committed suicide patients were found in total, and the average incidence rate per year in the recent decade was 30. 16/100 000. 5 attempted suicide patients were found and the average incidence rate per year in the recent decade was 4.71/100 000. Conclusion The Jinuo committed suicide incidence rate is higher than nationwide level. The attempted suicide incidence rate is lower than nationwide level.