1.Correlation between mutant mRNA expression of COL4A5 and phenotypes of XLAS females
Yunfeng WANG ; Jie DING ; Fang WANG
Chinese Journal of Nephrology 1997;0(01):-
Objective To understand the different phenotype based on COL4A5 mutations and to analyze the correlation between mutant mRNA expression of COL4A5 gene and phenotype variability of XLAS females. Methods Skin biopsy specimens were obtained from 6 XLAS females with confirmed COL4A5 deletion mutations. Total RNA was isolated from cultured skin fibroblasts and RT-PCR was performed to amplify the fragment including the mutation sequences of COL4A5 gene. The PCR products were electrophoresed with 8% polyacrylamide gel. Each fragment of PCR product revealed under polyacrylamide gel was further analyzed with the optical absorptance (A). The A ratio of mutant over total mRNA of COL4A5 gene (total mRNA means the mutant mRNA plus normal mRNA) was analyzed with different phenotypes. Results Besides microscopic hematuria, the females often had gross hematuria and persistent and severe proteinuria when the mutant a5 (IV) mRNA expression was more than the normal. Whereas the XLAS females didn't had gross hematuria and proteinuria or had discontinuous and mild proteinuria when the mutant ?5 (Ⅳ) mRNA expression was less than the normal. The proteinuria was positively correlated with the expression ratio of mutant over total mRNA of COL4A5 gene in the females. Conclusions The quantity of the mutant mRNA expression of COL4AS gene is correlated with the phenotypic severity of XLAS females. The XLAS females with much more mutant mRNA expression of COL4A5 gene show more severe phenotypes of AS.
2.Three Dimensional Contrast Enhanced Subtraction MRA:A New Technique in theDiagnosis of Body and Lower Extremity Blood Vessel Diseases
Xian DING ; Kai YANG ; Yunfeng ZHU ; Kuilun YU ; Zhongze ZHANG
Journal of Practical Radiology 2000;0(02):-
Objective To study the value of three dimensional contrast enhanced subtraction MRA(3D CES MRA)in the diagnosis of body and lower extremity blood vessel disease Methods Eighteen cases were studied by 3D ECS MRA and proved by operationResults 3D CES MRA images were of diagnostic quality without ghost for all 18 patients.3D CES MRA clearly showed normal vascular anatomy and various disorders.According to surgery,the sensitivity and specificity of 3D CES MRA in the diagnosis of body and lower extremity blood vessel diseases were both 100%.Conclusion 3D CES MRA is a new technique in diagnosing of body and lower extremity blood vessel diseases.It is proved to be a simple,noninvasive and commercial method in diagnosing blood vessel diseases.
3.Significance of amplitude integrated electroencephalogram in screening of neonatal seizures
Xinlin HOU ; Dandan ZHANG ; Yunfeng LIU ; Haiyan DING ; Congle ZHOU
Chinese Journal of Perinatal Medicine 2011;14(2):102-105
Objective To study the evaluative values of amplitude integrated electroencephalogram (aEEG) in screening neonatal seizures.Methods The aEEG and video electroencephalogram (VEEG) on 32 patients with seizures hospitalized in Peking University First Hospital was conducted on the same day.The parameters of aEEG including the integrated spectra band,background rhythm and primitive electro signal were analyzed and compared with VEEG.Results Seizures presented clinically by 12 cases were detected by VEEG,and their electric discharges were detected by aEEG.The sensitivity of aEEG was 100%.VEEG showed electric discharges during intermediate stage in twenty cases and thus the seventeen of this group were detected by aEEG with its sensitivity of 87% and 100% in the specificity.The positive and negative predictive values of aEEG were 80% and 100%,respectively.But aEEG could not discriminate the attack stage or intermediate stage electric discharges.The abnormal background activity was detected by VEEG in 13 cases and the sensitivity of aEEG was 100% and the specificity was 83%.The positive and negative predictive value by aEEG were 76% and 100% respectively.The electric discharges of 32 cases were detected by VEEG whenever they were in attack stage or intermediate stage.All electric discharges could be detected through primitive electro signal by aEEG.The electric discharges in 3 cases were not detected by integrated spectra band from aEEG,but were detected by primitive electro signal from aEEG.Conclusions The sensitivity and specificity in the screening of neonatal seizures by aEEG is useful and could be applied clinically as a simple and safe screening method for recognizing neonatal seizures.
4.IgM nephropathy with nephrotic syndrome in children
Zhihui LI ; Ying WANG ; Cuirong DUAN ; Tianhui WU ; Yunfeng DING
International Journal of Pediatrics 2015;42(3):334-337
Objective To investigate the clinical charactors and treatment of IgM nephropathy with nephrotic syndrome(NS) in children.Methods Thirty-six IgM nephropathy patients hospitalized in Hunan Children's Hospital as research group(group A),from June 2005 to June 2012.One hundred and six patients with minimal change disease (MCD) as control group (group B),followed up for 1 ~ 8 years.Results (1) Hematuria at presentation of the two groups respectly 3.8% vs 30.6% (x2 =20.403,P < 0.05).(2) Renal pathology revealed that there were 26 (72.2%)patients with minimal change disease,9 cases (72.2 %)with moderate membranoproliferative glomerulonephritis,1 cases with focal segmental glomerulosclerosis.(3) According to renal pathology,group A patients were divided into two sub groups:mild group and moderate group.To compared with group B,the steroid-resistant incidence of the 3 group were respectly 12.3%,19.2%,77.8% (x2 =24.369,P < 0.05).There was no significant difference between control group and mild group.(4)The remission rate of proteinuria in steroid-resistant patients who combined to use mycophenolate mofetil (MMF) with the two groups were respectively 50% and 85.7 % (x2 =3.60,P > 0.05).Conclusion Incidence of hematuria is higher in IgM nephropathy patients and patients with renal pathology for moderate lesions have a high steroid-resistant,and need use immunosuppressive early.MMF may be a good immunosuppressive for theses patients.
5.Clinical diagnosis and treatment for pancreatic injury in children
Xiaohong WANG ; Yunfeng DING ; Weimin ZHOU ; Fengyuan GU ;
Chinese Journal of Emergency Medicine 2006;0(07):-
Objective To summarize the clinical features of pancreatic injury in children,and to explore the suitable methods for its diagnosis and treatment.Methods A retrospective analysis of children with pancreatic injury within the 15 years was carried out.Clinical injury severity and following-up data were collected.Correlationship between injury severity,treatment method and successful rate was analyzed.Results Eleven children aged 4 to 14 years old were diagnosed as pancreatic injury:two had GradeⅠinjury,three GradeⅡ,five GradeⅢ,and one GradeⅣ.CT scan was used in 7 children,and 5 had positive sign.Patients with GradeⅠand GradeⅡinjuries were successfully cured without surgery.Three children with GradeⅢinjury were initially treated without operation,but two developed a large symptomatic pseudocyst and were cared through operation,and another one died.The other two children with GradeⅢwho underwent a distal pancreatectomy cured without complications.A patient with GradeⅣinjury was treated by Berne operation and died. Conclusion The management of pancreatic injury in children should be individualized depending on diagnosis time and injury type.A repetitive CT scan was helpful within 24h after injury.Distal pancreatectomy was a safe and effective for those with GradeⅢinjury.
6.Preparation of hepatoblasts from fetal liver tissues
Erjian ZHU ; Yunfeng PIAO ; Baijing DING ; Jing LI
Chinese Journal of Tissue Engineering Research 2007;0(05):-
BACKGROUND:Hepatoblasts are one of high-grade cell sources for severe liver disease treatment.Efficient separation and purification of hepatoblasts from fetal liver has important practical significances.OBJECTIVE:To introduce the preparation of hepatoblasts from fetal liver,conduct a transversal comparison of these preparation methods,and point out their advantages and shortcomings,so as to provide the evidences for the experimental and clinical choices of hepatoblasts.RETRIEVAL STRATEGY:Using the keywords of "fetal liver progenitor/hepatoblasts,isolation/purification",we searched the articles about fetal liver hepatoblasts separation and purification in PUBMED between January 1997 and August 2007 in English.At the same time,we searched the relative articles in CNKI using the keywords of "liver,embryo,endothelial progenitor/hepatoblasts" between January 1997 and August 2007 in Chinese.In addition,some related books and foreign articles were checked in the laboratory of Jilin University.After the first examination,the articles published in the authority magazines within five years were given prior consideration.Exclusive criteria were those repetitive researches or Meta-analysis.Totally 245 articles were collected,we selected 33 representative articles about hepatoblasts separation and purification methods.LITERATURE EVALUATION:Among 31 included articles,10 ones introduced the study background,5 ones studied the fetal liver cell dissociation,9 ones referred to the hepatoblasts separation and purification,and 7 ones indicated the problems and prospect in this field.DATA SYNTHESIS:The hepatic epithelial cell precursor cells(also known as hepatoblasts) with hepatic and biliary epithelial cell differentiation potential isolated from fetal liver,could in vitro proliferate and differentiate,after transplantation in vivo they show a good homing,and the ability to integrate and regenerate.Hepatoblasts isolation technology is being improved daily,and the related researches on this field will boost cell-based liver therapy in clinical application.CONCLUSION:Improvements and combination of the various isolation methods,as well as new and high-specific antigen discovery will further improve the isolation efficiency of hepatoblasts from fetal liver.
7.Expression of imprinted gene PEG10 in human gastric adenocarcinoma tissues and significance
Kai WANG ; Yunfeng PIAO ; Dayong DING ; Ye FENG
Journal of Jilin University(Medicine Edition) 2006;0(02):-
Objective To investigate the expression of a novel imprinted gene,PEG10,in human gastric adenocarcinoma,and the effect of PEG10 on cell growth and proliferation of gastric cancer cells.Methods The PEG10 mRNA expressions in 40 human gastric adenocarcinoma,the corresponding adjacent normal tissues and 6 nomal gastric tissues were detected by RT-PCR.The expression vectors of PEG10 were constructed and transfected into gastric cancer cell line MKN45 which had no endogenetic PEG10 expression.Cell growth ability was measured by MTT assay.Results High PEG10 mRNA expression level was detected in 9 of 20(45.0%) human gastric adenocarcinoma which was significantly higher than those of the matched normal tissues(10.0%)(P
8.Effects of efaroxan on insulin release from pancreatic βcells
Yi ZHANG ; Yunfeng LIU ; Jingying GAO ; Yaqin DING
Chinese Pharmacological Bulletin 2015;(4):524-526,527
Aim To study the insulinotropic effects of Efaroxan and the underlying mechanism in rat βcells. Methods Pancreatic islets were isolated by college-nase p digestion.Radioimmunoassay was used to meas-ure insulin secretion and cAMP level in rat pancreatic islets.Results Efaroxan only potentiated insulin se-cretion at high glucose concentrations(8.3,1 1 .1 mmol ·L -1 )but not at low glucose concentrations.KU1 4R,an antagonist of Efaroxan,remarkably inhibited Efarox-an-potentiated insulin secretion;and similarly,KU1 4R significantly inhibited forskolin-induced and IBMX-in-duced insulin secretion.cAMP measurement showed that forskolin and IBMX significantly increased cAMP levels,but Efaroxan and KU1 4R had no effects on cAMP content in pancreatic islets.Conclusion The mechanism of Efaroxan-potentiated insulin secretion is related to downstream of cAMP signaling pathway, KU1 4R antagonized the downstream of cAMP signaling leading to its inhibitory effects on Efaroxan,forskolin and IBMX-induced insulin secretion.
9.Expression of the partial protein encoded by mutated COL4A5 gene and analysis of the structure by circular dichroism
Yunfeng WANG ; Jie DING ; Dingfang BU ; Fang WANG ;
Journal of Peking University(Health Sciences) 2004;0(02):-
T mutation resulting in p.G1015V from an X linked Alport patient, and that of corresponding cDNA from a control were expressed in E.coli. The recombinant and mutant polypeptide was a fragment of COL4A5 , containing 158 amino acid residues with a glycine to valine substitution mutation in it. The secondary structure of the two recombinant proteins was analyzed using circular dichroism(CD) spectroscopy. Results: CD spectra of the control exhibited a negative peak near 200 nm whereas that of the patient exhibited a negative peak near 220 nm. The magnitude of the negative peak of the patient decreased as compared with that of the control. Furthermore, secondary structure of the control polypeptide was mainly composed of ? sheet and random coil without ? helix, whereas that of the patient presented 12.9% ? helix. Conclusion: Not only local structure of the substitution site but also folding kinetics of the entire ?5 chain may be changed due to Gly→Val substitution in Alport syndrome. We speculate that the abnormally folded polypeptide chain may not be assembled into the triple helix and the network of type Ⅳ collagen, or may be assembled into loosen triple helix then degraded easily, resulting in the pathognomonic ultrastructural changes of the glomerular basement membrane.
10.Regulatory Effects of miRNA-31 on LATS2 and Cardiomyocyte Hypertrophy in Rat's Cardiomyocytein vitro
Junyi ZENG ; Wan ZHANG ; Lu DING ; Yunfeng WEI ; Zeqi ZHENG ; Tong WEN ; Yongnan FU
Chinese Circulation Journal 2017;32(2):177-182
Objective: To observe the regulatory effects of miRNA-31 (miR-31) on LATS2 and cardiomyocyte hypertrophy via down-regulating miR-31 expression in rat's cardiomyocytesin vitro. Methods: Rat's cardiomyocytes were isolated and cultured for 10 daysin vitro, according to different intervention methods, the cells were divided into 4 groups:①Blank control group,②AngII intervention group,③Lentivirus with miR-31 inhibitor infection group,④Negative lentivirus infection group. On day-8, gene expressions of MiR-31, LATS2, cardiac hypertrophy ANP and β-MHC were examined by qRT-PCR; on day-10, cell morphology was observed by fluorescence staining. LATS2 protein expression was examined by Western blot analysis. Dual luciferase reporter plasmids were transfected into 293T cells, then luciferase activity was detected to identify the targeting effect of miR-31 on LATS2. Results: Compared with Blank control group, AngII intervention group showed increased gene expressions of miR31, cardiac hypertrophy ANP and β-MHC,P<0.05, enlarged cardiomyocyte surface,P<0.05; while decreased gene and proteinexpressions of LATS2,P<0.05. Compared with AngII intervention group, Lentivirus with miR-31 inhibitor infection group had down-regulated expressions of miR31, cardiac hypertrophy ANP and β-MHC,P<0.05, reduced cardiomyocyte surface, P<0.05; while slightly increased LATS2 gene expression and obviously increased protein expression,P<0.05. Dual luciferase reporter assay presented that relative luciferase activity of TRAF6-3' UTR+miR-146b was significantly decreased than TRAF6-3' UTR+miR-NC,P<0.01 and relative luciferase activity of LATS2-3' UTR+ miR-31 was signiifcantly reduced than LATS2-3' UTR-NC+miR-31,P<0.01. Conclusion: Cardiomyocytes hypertrophy could be reversed at certain degree by down-regulating miR-31; the targeting effect of miR-31 on LATS2 was involved in cardiomyocyte hypertrophyregulation.