3.Proteins structure changes of COL4A5 gene point mutations and structure-phenotype relations in Alport syndrome.
Yun-Feng WANG ; Jie DING ; Fang WANG ; Ding-Fang BU
Chinese Journal of Pediatrics 2004;42(8):589-594
OBJECTIVEAlport syndrome (AS) is a progressive hereditary nephritis presented with hematuria and renal failure, frequently associated with sensorineural deafness and ocular lesions. So far, more than 300 gene mutations in AS have been identified which provides a better way to analyze the association between genotype and phenotype. It is hard to understand all the phenotype according to the gene mutations, because the structure and function changes of the relevant protein, alpha5(IV) chain, encoded by mutated COL4A5 gene are rare to know. This study aimed to detect the proteins structure encoded by COL4A5 gene with different missense mutations and to analyze the effect of gene mutations on the secondary structure of alpha5(IV) chain and structure-phenotype relations.
METHODSTwo X-linked AS patients with different missense mutations (g.3246G > T resulting in p.G1015V and g.3290G > A resulting in p.G1030S, respectively) diagnosed by clinical manifestations, family history and skin or renal biopsy examinations, as well as a control were included in this study. The fragments of cDNA with the two mutations, respectively, and that of corresponding cDNA from the control were expressed in E. coli. The secondary structure of the recombinant polypeptides were analyzed by using circular dichroism (CD) spectroscopy.
RESULTSCD spectra of the control exhibited a negative peak near 200 nm whereas that of the patient 1 exhibited a negative peak near 220 nm. Furthermore, the magnitude of the negative peak of patient 1 decreased from -9000 deg x cm2 x dmol(-1) to -4000 deg x cm2 x dmol(-1) as compared with that of the control. CD spectra of the patient 2 were slightly changed with the negative peak remaining near 220 nm but the magnitude increasing from -9000 deg x cm2 x dmol(-1) to -11000 deg x cm2 x dmol(-1) as compared with that of the control. In addition, the secondary structure of the control polypeptide was mainly composed of beta-sheet and random coil without alpha-helix, whereas that of the patient 1 presented 12.9% alpha-helix. Although the secondary structure of polypeptide of the patient 2 was also mainly composed of beta-sheet and random coil, the composition of beta-sheet reduced and random coil increased.
CONCLUSIONAlthough the glycine substitutions existed in the same domain of alpha5(IV) chain, the patient 1 with the severe AS phenotype and g.3246G > T mutation, and patient 2 with the mild AS phenotype and g.3290G > A mutation were revealed with different secondary structures of alpha5(IV) chain. Moreover, the secondary structure changes of alpha5(IV) chain were consistent with their corresponding phenotype severity.
Collagen Type IV ; genetics ; Humans ; Mutation, Missense ; Nephritis, Hereditary ; genetics ; Phenotype ; Point Mutation ; Protein Structure, Secondary ; genetics ; Spectrum Analysis
5.Measurement of the mRNA level of Polo-like kinase 1 in bronchoscopic bioptic specimens by realtime quantitative PCR
Ke-Jing YING ; Fang-Chun SHAO ; Bi-Yun YU ; Jin-Min WU ; Jia-Yi DING ;
Chinese Journal of Laboratory Medicine 2003;0(09):-
0.05)and the TNM staging (P=0.55).A mild elevated compared other pathological classification was found in small cell lung cancer (0.191?0.275).Conclusions The results showed that RFQ-PCR was suitable for measurement of the mRNA level of PLKI in bronchoscopic bioptic specimens.This study suggest elevated expression of PLK1 might play a important role in development of lung cancer,so that PLK1 might be a potential tumor marker for Lung cancers.Advanced studies will be needed to clarify that PLKI mRNA level do not relate to TNM staging and pathological classification.
6.Spatial distribution of wall shear stress in the common carotid artery of type 2 diabetes mellitus patients
Zhifang HUANG ; Ming CHEN ; Yuhui ZHANG ; Yun DONG ; Ying WEI ; Fang DING ; Qiuyun LIU
Chinese Journal of Ultrasonography 2014;23(5):393-397
Objective To investigate the wall shear stress(WSS) in the common carotid artery of type 2 diabetes mellitus(DM) patients,and analyzed the spatial distribution of WSS by using quantitative visualization of blood flow shear stress analysis software.Methods Eighteen male type 2 DM subjects were enrolled as DM group and 18 age-matched healthy subjects were selected as control group.None of the participants was hypertensive,hyperlipidemic or a cigarette smoker.Intimal-medial thickness (IMT),number and size of plaques in the common carotid artery were evaluated by high-resolution echo-Doppler.Color Doppler flow images of common carotid arteries in the two groups were extracted from DICOM files.WSS in the common carotid arteries was calculated by shear stress visualization quantitative analysis software,and the corresponding spatial distribution maps of WSS were designed.Results WSS of the common carotid arteries in the control group were ranged from 4 to 14 dyne/cm2.WSS of the common carotid arteries in the DM group were ranged from 2 to 8 dyne/cm2.Compared to mean WSS value [(6.96 ± 1.17)dyne/cm2] of common carotid arteries in the control group,mean WSS value [(3.14 ± 0.79)dyne/cm2] of common carotid arteries in the DM group was significantly lower (t =9.380,P =0.000).Six diabetic participants had a plaque in one carotid artery and no lesions in the contralateral carotid.Among these subjects,mean WSS was significantly lower in the side with lesion (t =7.324,P =0.000).Therefore,IMT of common carotid arteries in participants was significantly inversely related to WSS (r =-0.76,P <0.01).Conclusions The common carotid arteries of DM patients are more prone to atherosclerosis which is associated with reduction of WSS.The hemodynamic profile might represent an additional factor contributing to the increased prevalence and severity of carotid atherosclerosis in diabetic patients compared with general population.
7.In vivo interleukin-10 gene transfer down-regulates myocardial matrix metalloproteinase and myocardial collagen expressions in rats with acute myocardial infarction.
Chun-Yang HU ; Wen-Hui DING ; Xiao-Ning HAN ; Song-Yun CHU ; Yan-Jie HAO ; Ding-Fang BU
Chinese Journal of Cardiology 2008;36(3):243-248
OBJECTIVEWe investigated the in vivo effects of recombinant adenovirus-associated virus type-2 (AAV-2) mediated interleukin-10 (IL-10) gene transfer on the expression of matrix metalloproteinase (MMP)-2, 9, tissue inhibitor of metalloproteinase (TIMP)-1, collagen type I and type III in a rat acute myocardial infarction model.
METHODMale Sprague-Dawley (SD) rats were randomly divided into three groups (each n = 6): sham operation group, MI/AAV2 group, and MI/AAV2-IL-10 group (10(10) vg/ml x 0.1 ml injection at peri-infarct regions immediately post MI). Five days later, the expressions of MMP-2 and MMP-9 were measured by RT-PCR, Western blot and zymography. The expression of TIMP-1 was measured by RT-PCR and Western blot. Collagen type I and type III were assessed by RT-PCR and immunohistochemical stain.
RESULTSThe myocardial expressions of MMP-2, MMP-9 and collagen contents in MI/AAV2 group were significantly increased than those in sham operation group. Myocardial expressions of MMP-2, MMP-9 were significantly decreased and the expression of TIMP-1 significantly increased in the MI/AAV2-IL-10 group than those in MI/AAV2 group. Moreover, the expressions of collagen type I, collagen type III and the ratio of I/III collagen in border zones of infarcted myocardium were decreased by 47.6% (P < 0.01), 23.6% (P < 0.05), and 17.9% (P < 0.05) respectively, while the expression of TIMP-1 increased by 73.1%(P < 0.05) in MI/AAV2-IL-10 group compared to MI/AAV2 group.
CONCLUSIONIn vivo myocardial IL-10 transfer reduced myocardial MMP and collagen expression and increasing the TIMP expression.
Animals ; Extracellular Matrix ; metabolism ; Gene Expression ; Genetic Therapy ; Interleukin-10 ; genetics ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Myocardial Infarction ; genetics ; metabolism ; physiopathology ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism ; Transfection ; Ventricular Remodeling
8.Clinical trial on treatment of acute cerebral infarction with TCM treatment according to syndrome differentiation combining Western medicine by staging.
Ding-Fang CAI ; Yun-Ke YANG ; Xi-Xi GU
Chinese Journal of Integrated Traditional and Western Medicine 2007;27(9):789-792
OBJECTIVETo observe the curative effect of integrated Chinese and Western medicine on acute cerebral infarction (ACI).
METHODSTwo hundred and seventy-nine ACI patients were assigned to two groups. The control group (140 cases) was treated with Western medicine by staging and the treated group (139 cases) was given TCM therapy according to syndrome differentiation on the basis of Western medicine. The end point was set at the 90th day of the administration. The curative effect was assessed by Chinese stroke scale (CSS), National Institutes of health stroke scale score (NIHSS), Rankin scale and Barthel index (BI).
RESULTSAt the end point of the trial, the total effective rate was 73.38% in the treated group and 61.43% in the control group, the former was superior to the latter (P<0.05). There were 66 cases (47.14%) in the control group and 80 cases (57.55%) in the treated group with improvement rate of NIHSS > or =40%, 65 cases (46.43%) with their Rankin scale within 0-2 grade in the control group and 78 cases (56.12%) in the treated group, 60 cases (42.86%) in the control group and 71 cases (51.08%) in the treated group with BI > or =85, 61 cases (43.57%) in the control group and 72 cases (51.80%) in the treated group with improvement rate of CSS > or = 46%, comparison between them showed significant differences (P < 0.05).
CONCLUSIONTCM therapy accord-ing to syndrome differentiation combined with Western medicine by staging shows better curative efficacy on ACI.
Acute Disease ; Adult ; Aged ; Anticoagulants ; therapeutic use ; Cerebral Infarction ; diagnosis ; drug therapy ; Diagnosis, Differential ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Phytotherapy ; Syndrome ; Treatment Outcome
10.An Intelligent Search Engine System Application for Disinfection Protection:Investigation Guiding
Jian FANG ; Xin-Jian LIU ; Ding-Hua SHEN ; Hong LV ; Jie ZHU ; Jing-Ya MIAO ; Li MA ; Yun SHAO ;
Chinese Journal of Nosocomiology 2004;0(10):-
OBJECTIVE To develop a non-paper,interactive,multiplex medium way,and to obtain the information disinfection protection on second level of time.METHODS Using the computer,the touchscreen,the personal digital assistant(PDA),the general packet radio service(GPRS),the pronounciation(man-machine conversation),the video frequency and the other advanced technology methods,to study and develop an electronic version of intelligent search engine system for disinfection protection.RESULTS Touching namely attained the applied and guided information,such as to fulfil exactly the disinfection technology standard for the medical workers in medical establishments,to provide the post work instruction;to correctly guide the disinfection protection for common family;and to build the communication platfrom with the enterprisers in order to meet the demand for disinfection protection appliances from the society,hospital and family.CONCLUSIONS The intelligent search engine system is feasible applying to use.