1.Bailing Capsule in Preventing Epithelial- Mesenchymal Transition in Rats with Tubulointerstitial Fibrosis
De-an, ZHAO ; Da-sheng, YANG ; Ling-yun, BI ; Hui-qin, XU ; Ying, ZHAO ; Xin-de, CHEN
Journal of Applied Clinical Pediatrics 2005;20(9):939-942
Objective To explore the effect of Bailing capsule on epithelial-mesenchymal transition( EMT) in rats with adenine-in-duced tubulointerstitial fibrosis. Methods Tubulointerstitial fibrosis animal models were established and SD rats were divided into mo-del group ( n = 30), treatment group ( n = 30) and control group( n = 30), randomly. Experimental rats were harvested at 7 w, 12 w,17 w after onset of experiment and functional evaluations were performed. Histology, immunohistology were examined to investigateboth histolopathology changes and the expression of bone morphogenic protein-7 (BMP-7), transforming growth factor-β1 (TGF-β1 )and a-smooth muscle actin (α-SMA) in kidneys at three time points mentioned above, respectively. Results Compared with controlgroup, 24 h urinary protein in model group lost increasingly and significantly difference appeared at three time points relative to controlgroup ( P < 0.01 ). Urinary NAG in model group was markedly higher than that in control group from 7 w after onset (P < 0.01 ) andwas increasingly raised at 12 w and 17 w (P<0.01). The value of blood BUN and Cr in model group increased at 7 w (P>0.05) rel-ative to control group. There was significant difference at 12 w and 17.w (P < 0.01 ). Histologically, kidneys in model group, at 7 w,exhibited tubular casts and gently tubular dilation, granuloma in cortex, mononuclear cells infiltration in tubulointerstitial areas, andmild interstitial fibrosis. At 12 w, the degree of tubular injury and tubulointerstitial fibrosis gradually aggravated. Up to 17 w, diffusetubular dilation or atrophy was observed and focal tubules disappear. Diffuse interstitial fibrosis was exhibited. In normal kidneys, im-munohistochemistry suggested that the light expression of BMP-7 was detected in proximal renal tubular epithelial cells and marked ex-pression was identified in distal tubule, collecting duct, and renal tubular epithelial in junction area between cortex and medulla. How-ever, the expression of BMP-7 in kidneys of model group significantly decreased with increasing tubulointerstitial fibrosis and was nega-tive correlation with the expression of TGF-β1(r = -0. 981 P<0.01) and α-SMA (r= -0.975 P<0.01). Bailing capsule ad-ministration protected the expression of BMP-7 and reduced TGF-β1 and α-SMA expression before 12 w(P< 0.01 ). Conclusions Ourstudy shows an anti-fibrotic reno-protective function of Bailing capsule in rats with tubulointerstitial fibrosis via prevention of epithelial-mesenchymal transition at early stage. However, the beneficial effect lost with increasing tubulointerstitial fibrosis.
2.Bailing Capsule in Preventing Epithelial -Mesenchymal Transitionin Rats with Tubulointerstitial Fibrosis
de-an, ZHAO ; da-sheng, YANG ; ling-yun, BI ; hui-qin, XU ; ying, ZHAO ; xin-de, CHEN
Journal of Applied Clinical Pediatrics 2004;0(09):-
Objective To explorethe effect of Bailingcapsule on epithelial-mesenchymal transition(EMT) inrats withadenine-in-duced tubulointerstitial fibrosis .Methods Tubulointerstitial fibrosis ani mal models were established and SDrats were dividedinto mo-del group (n=30) ,treatment group (n=30) andcontrol group(n=30) ,randomly .Experi mental rats were harvested at 7 w,12 w,17 wafter onset of experi ment and functional evaluations were performed. Histology ,i mmunohistology were examined to investigateboth histolopathology changes and the expression of bone morphogenic protein-7 (BMP-7) ,transforming growth factor-?1(TGF-?1)and a-smooth muscle actin (?-SMA) in kidneys at three ti me points mentioned above ,respectively .Results Compared with controlgroup ,24 h urinary proteinin model grouplost increasingly and significantly difference appeared at three ti me points relative to controlgroup(P0 .05) rel-ative to control group.There was significant difference at 12 wand 17 w(P
3.Observation of the Expression of HCV NS 5 Antigen in vitro by the SABC Immunological Techniques and Gold-labeled Colloid Electron Microscopy Method
Jin, YE ; Ling-lan, ZENG ; Mu-lan, YANG ; Duan-de, LUO ; Jin-song, GUO
Virologica Sinica 2001;16(1):88-91
To study the expression of HCV non-structure 5 antigen in vitro, a human HepG2 cell line was incubated with a HCV RNA positive serum. The S ABC i mmunological techniques and gold-labeled colloid electron microscopy method wer e employed to examine for the viral proteins in those cells. The HCV non-struct ure 5 antigen was first detected in the HepG2 cells at 72 hours post incubation. The antigen was continuously observed in the cytoplasm or on the membrane as we ll on the cell wall of the HepG2 cells even after 1, 2, 3 and 4 weeks post incub ation. The observation of HCV non-structure 5 antigen continuously expressed in the HepG2 cells strongly indicates that the cells may have been infected by HCV virus and the virus may have replicated in the cells. Therefore, the HepG2 cell line may be served as a potential host for establishment of HCV infection and p ropagation in vitro.
4.Effects of nitric oxide,endothelin-1, and liver sieve on biliary fibrosis in rats
Wenjun YANG ; Zhengming LEI ; Ling MA ; Daiyu LI ; Jing LI ; De SHI
Chinese Journal of General Surgery 2001;0(10):-
Objective To study the effects of nitric oxide (NO) and endothelin-1(ET-1) on cholestasis liver fibrosis. Methods Forty-eight Wistar rats were randomily divided into 3 groups:the control group,the common bile dute ligation(CBDL)group,and the CBDL plus lactulose group.The animals were killed on the 3rd,7th,14th, and 21th days respectively to determine the contents of plasma endotoxin,NO,ET-1, and transaminases(ALT,AST).The degree of hepatic fibrosis was observed by microscopy and the ultrastructural changes of liver were observed by electron microscope. Results After common bile dute ligation,the contents of plasma ET-1 and NO increased significahtly,the plasma endotoxin level also increased and there were positive correlation between the levels of ET-1,and NO and the level of endotoxin.Plasma transaminases also increased greatly compared with those of the control group.The hepatic sinusoidal steonsis and widen Disse space were found.The diameter and number of fenestrate on hepatic sinusoid greatly decreased.Treatment with lactulose could partially alleviate all the pathological changes noted above. Conclusions NO and ET-1 may damage the "liver sieve"to cause the cholestasis liver fibrosis.
5.A study of histopathological changes of small bowel allograft during acute rejection in rats
Shicheng LUO ; De SHI ; Hui YANG ; Ling LI ; Jun ZHU ; Jianhu GUO
Chinese Journal of General Surgery 2000;0(11):-
Objective To investigate the features and significance of pathologic changes in apoptosis of small bowel allograft during acute rejection in rats. Methods All 24 recipients were equally divided into four groups ; group A: nonoperative control; group B: allograft ; group C: isograft, group D: treatment control. The graft samples were harvested on day 3, 5, 7, 10 after transplantation, and subjected to histologic examination . Mucosal thickness, villous height and crypt depth were measured, and apoptotic cells of intestinal mucosa of grafts on day 3,5 and 7 after transplantation were examined. Results The mucosal structure was normal in group A; The degree of the inflammatory infiltrated cells ,intestinal mucosa cell apoptosis and structural injury of mucosa in group B were significantly severe compared with groups C and D. As the post-transplanted time increased, the number of musocal apoptotic cells and the degree of mucosal structural injury were significantly increased. The degree of mucosal structural injury in group C was milder than in group B. A few infiltrated cells and mild edema of mucosa occurred in group D , but no mucosal structural injury was found. Conclusions Inflammatory cell infiltration, mucosal epithelial cell apoptosis and mucosal structural damage are the main pathologic features of small bowel allograft during acute rejection. Dynamic observation of the pathologic changes and cell apoptosis of small bowel graft is of certain value in the diagnosis of acute rejection of small bowel graft and in assessment of the degree of small bowel injury.
6.CLONING OF NATTOKINASE GENE AND EXPRESSION IN E. COLI
Li-Xin LUO ; Zhi-Li HUANG ; Ru-De YANG ; Jun-Jian LING ; Shi-Zhong LIANG ;
Microbiology 1992;0(03):-
In this study, nattokinase gene was amplified by PCR using bacillus subtilis chromosomal DNA as template and cloned into expressed vector pBV220. After transforming recombinant plasmid into E.coli HB101, the recombinant strain was yielded. It was proved that expression products was secretive and expression protein was 12% of total cell protein by SDS-PAGE. Optimum culture time and inducing time was determined as 6h and 5h respectively. The plasmid stability studies showed that recombinant plasmid has excellent segregational stability but the structural stability was not good in the host cell.
7.Isolation,Identification and Degradation Characteristics of a DMP-degrading Strain
De-Cai JIN ; Xue-Ling WU ; Ren-Xing LIANG ; Qin-Yun DAI ; Yang-Yang WANG ; Yu YANG ;
Microbiology 2008;0(09):-
A bacterial strain which could grow well on the substrate of PAEs as the sole source of carbon and energy was isolated from contaminated sludge in the river of WeiFang in ShangDong province and it was designated as JDC-3. Based on the morphology,biophysical and biochemical properties as well as molecular characteristics,this isolate was preliminarily identified as Delftia sp.. A fragment of phthalate dioxygenase gene was successfully amplified from the genus of Delftia for the first time using a set of degenerate primers. Meanwhile,the degradation capability of JDC-3 was determined by HPLC using DMP as test substrate. The results showed that the optimal pH and temperature were at 7.0~8.0 and 30?C~35?C respectively. The degradation kinetics of JDC-3 was studied in different initial DMP concentration under optimal conditions. The results indicated that the degradation dynamic equation was ln C =-0.06837 t + A when DMP concentration was lower than 300 mg/L,with half life of 12.48 h. The degradation rate decreased and half life of JDC-3 prolonged as the initial concentration kept on increasing.
8.Effects and Safety of Hemoperfusion Therapy on Infants with Tetramine Poisoning
de-xuan, WANG ; rui-xia, LIN ; qing, YANG ; guo-quan, PAN ; chun-xue, YAN ; ling-wei, JIN
Journal of Applied Clinical Pediatrics 1992;0(06):-
Objective To explore the effects and safety of hemoperfusion(HP) therapy on tetramine poisoning in infants.Methods Thirty-five infants with tetramine poisoning were divided into two groups: HP group(n=18) and non HP group(n=17).The changes of blood tetramine concentration and clinical symptom improving of both groups after the treatment were observed together with the adverse effects of HP group.Results The average blood tetramine concentration of HP group was higher than that of non HP group(342.2?333.4 vs 117.9?50.8 ?g/L,P
9.Relationship between the polymorphism of lipoprotein lipase gene and serum lipid levels in the Guangxi Hei Yi Zhuang population
Yong WANG ; Rui-Xing YIN ; Guang-Qin CHEN ; Wei-Xiong LIN ; De-Zhai YANG ; Shang-Ling PAN ;
Chinese Journal of Laboratory Medicine 2003;0(09):-
0.05 for all).Conclusions The allelic frequencies of LPL gene at Pvu Ⅱ locus in Hei Yi Zhuang were different from those in Han,but the genotypie frequencies in Hei Yi Zhuang were not different from those in Han.There was no significant correlation between the polymorphism of LPL gene at Pvu Ⅱ site and the serum lipid levels in two ethnic groups.
10.MicroRNA alterations in senescent endothelial progenitor cells induced by remnant-like lipoproteins.
De-Guang YANG ; Ling LIU ; Sheng-Hua ZHOU
Chinese Medical Journal 2012;125(19):3479-3484
BACKGROUNDRemnant-like lipoproteins (RLPs) have been demonstrated to accelerate the onset of endothelial progenitor cells (EPCs) senescence. Recent study has determined that microRNAs (miRNAs) were closely associated with cellular proliferation and senescence. This study aimed to examine whether RLPs lead to an alteration of miRNAs in senescent EPCs.
METHODSRLPs were prepared from plasma samples with immunoaffinity method. After 8 days of culture, EPCs were identified by flow cytometry analysis. Cells were incubated with RLPs for 72 hours. The senescent markers p16INK4a and senescence-associated beta-galactosidase (SA-β-gal) were detected by Western blotting analysis and β-gal staining assay, respectively. A human miRNA microarray containing 723 miRNAs was used to detect the expression profile of miRNAs in control and senescent EPCs. The result from the above microarray was qualified by RT-PCR assay.
RESULTSRLPs dose-dependently up-regulated the protein level of p16(INK4a) in EPCs, and RLPs at a concentration of 100 µg/ml induced a significant increase in the percentage of SA-β-gal-positive EPCs. Of 723 miRNAs, four miRNAs expressed differentially and significantly in RLPs-treated EPCs (P < 0.05), then their changes in expression were validated by real-time RT-PCR. Among them miR-148b and miR-155 were upregulated while miR-574-3p was down-regulated significantly when compared with control (P < 0.01).
CONCLUSIONSRLPs result in the onset of EPCs senescence. Senescent EPCs induced by RLPs exhibit a different profile of miRNAs. These three miR-148b and miR-155 and miR-574-3p reach a significant difference when compared with control, indicating that microRNA might take part in RLPs-induced EPCs senescence.
Blotting, Western ; Cells, Cultured ; Cellular Senescence ; drug effects ; genetics ; Cyclin-Dependent Kinase Inhibitor p16 ; metabolism ; Endothelial Cells ; cytology ; drug effects ; Flow Cytometry ; Humans ; Lipoproteins ; pharmacology ; MicroRNAs ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cells ; cytology ; drug effects