1.Dynamic relationship between macrophage and extra cellular matrix in the kidney of rats with experimental nephritis
Changwu WAN ; Bing GUO ; Guozhong ZHANG ; Huazhen GUI ; Yali ZHANG
Chinese Journal of Pathophysiology 2000;0(08):-
AIM: To observe the changes in infiltrating macrophages (M?)and extracellular matrix (ECM) in the kidney in the progressive course of nephrotoxic nephritis (NTN). METHODS: NTN model was established with rabbit-anti-rat nephrotoxic serum. On day 3, 7, 15, 30 and 90, renal biopsies were performed. Renal histology was checked under light microscopy with HE and Masson's trichrome staining sections. M?, fibronectin (FN), type Ⅲ and Ⅳ collagen were examined with immunohistochemistry ABC method. RESULTS: Infiltration of M? appeared on day 3 of NTN and preceded changes of FN and collagen. On day 15 of NTN, all nephritic animals had significant proteinuria, increased serum creatinine, infiltration of M? and deposit of entracellular matrix. On day 90 of NTN, seven nephritic animals improved significantly, while other five developed renal scarring with diffuse infiltration of M? which positively paralleled to renal function and deposit of FN, type Ⅲ and Ⅳ collagen. CONCLUSION: M? infiltrating into renal tissue enhances deposit of ECM and therefore plays important roles in progression or improvement of NTN.
3.Analysis of the effects of miRNAs on the expression of cytokines by THP-1 cells using Luminex xMAP technology
Lian GUI ; Qianqian ZHANG ; Yan CAI ; Qi GUO ; Junqi HUANG
Chinese Journal of Microbiology and Immunology 2015;(11):799-805
Objective To investigate the effects of a miRNA family member, let-7e, and a combi-nation of miR-106b and miR-20a on the expression of cytokines by THP-1 cells with Luminex xMAP technol-ogy.Methods The efficiency of transfection was evaluated by immunofluorescence assay after transfecting THP-1 cells with micrONTM mimic negative control (Cy3) for 24 h, 36 h and 48 h.The three miRNA mim-ics (let-7e, miR-106b and miR-20a) were respectively used to transfect the THP-1 cells for 24 h, 36 h and 48 h and the expression of each miRNA was analyzed by qRT-PCR analysis for screening out the optimal transfection time.The transfected THP-1 cells were stimulated with1 mg/L of LPS for 1 h.The Luminex xMAP technology was used to detect the expression of IL-8, interferon-inducible protein-10 (IP-10), mono-cyte chemotactic protein 1 (MCP-1), IL-1α, IL-6, IL-10, TNF-α, IFN-αand IFN-βin the supernatants of cell culture.A statistical analysis was performed to analyze the data obtained by using SPSS16.0 software. Results More than 90% of the transfected THP-1 cells were labeled with red fluorescence.The optimal transfection times for let-7e mimic and miR-106b/miR-20a mimics were 48 h and 24 h, respectively.Com-pared with the corresponding negative control (NC), the expression of IL-8, IP-10 and MCP-1 by THP-1 cells were enhanced after the transfection with let-7e mimic, but were inhibited after the co-transfection with miR-106b and miR-20a mimics.Conclusion The expression of IL-8, IP-10 and MCP-1 were enhanced in let-7e transfected THP-1 cells, but were inhibited in miR-106b and miR-20a co-transfected THP-1 cells.
4.Study on relationship between expression of PKC? in glomeruli and development of nephropathy in diabetic rats
Zhonghui JIA ; Guozhong ZHANG ; Bing GUO ; Huazhen GUI ; Changw WAN
Chinese Journal of Pathophysiology 1999;0(09):-
AIM: To observe the dynamic changes of expression of PKC? , TGF-? 1 and ?-SMA in glomeruli of diabetic rats induced by the alloxon and to invesitigate their roles in the diabetic nephropathy(DN). METHODS: Rats were randomly divided into four groups: normal control group (group A), diabetic group of one week (group B), diabetic group of one month (group C), diabetic group of two months (group D). Immunohistochemistry and Western blotting were used to detect the expression of PKC?, TGF-? 1 and ?-SMA in renal tissue of all groups. Blood glucose, triglycerides, cholesterol, creatinine and urine protein were analysed by chemical methods. The morphological changes of renal tissue were checked through microscopy. RESULTS: The expression of PKC? and TGF-? 1 in renal tissue of diabetic groups were increased comparing with those of nomal control group( P
5.Dynamical observation of the expression of TGF-?_1 and MAPK_(1/3) in the renal tubules of rats with diabetes
Bing GUO ; Ying XIAO ; Changwu WAN ; Huazhen GUI ; Guozhon ZHANG
Chinese Journal of Pathophysiology 1999;0(09):-
AIM: To observe the expression of transforming growth factor ? 1 (TGF-? 1), MAPK 1/3 and fibronectin (FN) in the development of renal tubulointerstitial disease. METHODS: Wistar male rats were randomly divided into normal control group, diabetic group of 1week, 2 weeks, 4 weeks and 8 weeks. Diabetic model was induced by peritoneal injection of streptozotocin. Immunohistochemistry was employed to detect the expression of TGF-? 1, MAPK 1/3 and FN in the kidney. TGF-? 1 protein in the renal cortex was checked by Western blot. BG, Scr and UP were analysed by biochemical methods, and the morphological changes in renal tubulointerstitium were also examined under microscopy on sections stained with HE and PAS. RESULTS: The expression of MAPK 1/3 and FN was observed, but not the expression of TGF-? 1 in normal renal tissue. Positive staining of TGF-? 1 was observed in the renal tubulo-interstitium in 1-week diabetic group and thereafter it increased in the course of diabetes. A continuous increase in the expression of MAPK 1/3 and FN was also observed in two - week diabetic rats. Chronologically the expression of TGF-? 1,MAPK 1/3 and FN and the ratio of KW/BW were positively correlative with each other in diabetic animals except one -week diabetic rats. There was also a positive correlation between MAPK 1/3 and FN in l -week diabetic rats. CONCLUSION: Our data suggest that TGF-? 1 appears in the renal tubulointerstitium in early period of diabetes and then its signal is mediated by MAPK 1/3 cascades to accelerate production of FN ,and in turn leads to renal hypertrophy and tubulointerstitial fibrosis. [
6.Effect of antisense transfection of monocarboxylate transporter on cell biological characteristics in human lung adenocarcinoma cells
Gui-Zhi ZHANG ; Gui-Jun HUANG ; Xian-Jian GUO ; Gui-Sheng QIAN
Journal of Third Military Medical University 2001;23(2):128-130
Objective To study the effect of transfecting anti-sense expression vector of the first subtype of the monocarboxylate transporter (MCT1) gene into human lung adenocarcinoma cells on intracellular pH (pHi) regulation, lactate transportation and cell growth. Methods MCT1 antisense gene recombinant vector pLXSN-MCT1 was introduced into human lung cancer cells A549 with electroporation. The cell colonies resistant to G418 were selected. Positive clones were examined by PCR to confirm the integration of genomic A549 DNA and antisene MCT1 gene. The changes of pHi and lactate transportation were detected with spectrophotometry. Cell growth was studied with cell growth curve. Results pHi and lactate transport were remarkably decreased in the transfected cells, and the cell growth was inhibited compared with the cells without transfection(P<0.001). Conclusion MCT1 gene may play an important role in pHi regulation, lactate transport and cell growth in lung tumor cells.
7.The inhibitory effect of decorin on proliferation of rabbit epithelial cell
Jian-nan, XIANG ; Gui-lan, ZHANG ; Hai-jiang, ZHANG ; Guo-hua, WANG ; Ming, HUO
Chinese Journal of Experimental Ophthalmology 2012;30(1):41-45
BackgroundResearches found that the posterior capsular opacification (PCO) after lensextraction is associated with the elevation of the transforming growth factor-β(TGF-β).To seek the drug for inhibitingproliferation of lens epithelial cells (LECs) is crucial in the treatment and prevention of PCO.ObjectiveThisstudy was to investigate the preventing effects of decorin on the proliferation of LECs.MethodsRabbit LECs wascultured and passaged.The LECs in growth phase were incubated in 96 well plate at the density of 8×106/L.Decorinwith the concentrations 0.1,1.0,10.0 mg/L was added into the medium for 24,48 and 72 hours respectively.0.1%DMSO was used at the same way as positive control group,and the regular cultured cells worked as blank controlgroup.The inhibitory rates of different concentrations of decorin on the growth of LECs were detected by MTT at 24,48and 72 hours after addition of decorin.The percentage of LECs in different cell cycles in various groups was assayedusing flow cytometry.TGF-β level in medium suspension was detected using ELISA.The expression of TGF-β mRNA in LECs was checked by RT-PCR,and α-SMA expression in LECs was determined using immunochemistry.Results ELISA assay showed a statistical difference in the TGF-β levels of different groups (F=39.24,P=0.03 ).The TGF-β levels in 1.0,10.0 mg/L decorin groups were significantly decreased in comparison with blank control group (P<0.01) and 0.1 mg/L decorin group (P<0.05 ).The inhibitory rates of decorin in the concentrations of ≥ 1.0 mg/L on the growth of LECs were higher than the blank control group,and those in various concentrations of decorin groups were considerably lower in 24 hours compared with 48 and 72 hours ( P<0.05 ) and so was the 48 hours compared with 72 hours (P<0.05 ).The percentages of LECs in G0/G1 phase were ascent in 0.1,1.0 and 10.0 mg/L decorin groups in comparison with G2/M and S phase (P<0.05).Immunochemistry revealed the weak expression of α-SMA in various decorin groups in comparison with control group. Conclusions Decorin can effectively inhibit LECs growth and induce LECs apoptosis in concentration- and time-dependent manner.It is suggested that decorin can be used in the prevention and treatment of after cataract.
8.Time-dependent change in expression of type-Ⅰ collagen protein gene during the development of restenosis after arterial injury
Ya-Wen ZHANG ; Guo-Yuan ZHANG ; Zong-Gui WU
Academic Journal of Second Military Medical University 2001;22(2):133-137
Objective: To observe the alteration of type- Ⅰ collagen protein gene expression after arterial injury and investigate its effect on the development of restenosis. Methods: Firstly, thee xperimental carotid arterial injury rabbit model was constructed. Then, Norther n blot, in situ hybridization and histomorphometric analysis were used to de tect the expression of procollagen mRNA and the accumulation of collagen protein 1,2,4 weeks after injury. Results: Type- Ⅰ collag en mRNA increased 1 week after injury, peaked 2 weeks later and decreased 4 week s later. The deposition of the collagen protein account for a high percentage o f space in neointima on histomorphometric analysis. Conclusion: Collagen protein may play an important role in the development of neointima and restenosis.
9.Temperature Sensation Threshold of Trunk Skin in Healthy Adults
Hui-li ZHANG ; Ming-ming GAO ; Hua-zhen GUO ; Gui-yun SONG ; Pu ZHANG
Chinese Journal of Rehabilitation Theory and Practice 2015;21(7):804-806
Objective To measure the temperature sensation threshold of trunk skin in healthy adults. Methods The threshold of cold sensation, warm sensation, cold pain sensation and heat pain sensation of trunk skin key points (T3, T7 and T11) were measured with Thermal Sensory Analyzer in 123 healthy adults. Results The thresholds of cold, warm, cold pain and heat pain sensations were obtained. The standard deviation of cold and warm threshold was less than that of heat pain. The range of cold sensation threshold was the largest. The heat pain sensation threshold increased with segmental declining and the sensation threshold increased with age. Conclusion Normal reference value should be established variously with the segment and age. The threshold of cold, warm varies less, while the threshold of cold pain and heat pain varies too much.
10.Model index observations in SIVmac251-infected rhesus macaques.
Yu ZHANG ; Jing WANG ; Xiang-mei LIU ; Fan-gui MIN ; Peng-jv GUO ; Ren HUANG
Chinese Journal of Virology 2014;30(6):675-682
In this study, five rhesus macaques were inoculated intravenously with SIVmac251 to establish a model of simian autoimmune deficiency syndrome (SAIDS). Peripheral blood samples were collected at different time points to monitor changes in the total T cell number and T lymphocyte subset. Plasma viral loads, cytokine expression levels and anti-SIV antibody levels were also assayed to acquire certain basic indexes to evaluate disease progression in the rhesus macaque SAIDS model. During the acute stage of infection, plasma viral loads reached a peak at week 1 post-inoculation and lasted for approximately 3 to 44 weeks. The CD3+ CD4+ T lymphocyte count in peripheral blood also transitorily decreased. During the same period, the level of interferon-gamma show an increasing trend, whereas IL-12 levels decreased; IL-2, IL-4, IL-10 and TNF-alpha were maintained at normal levels or could not be detected. During the asymptomatic and ARC phases, plasma viral loads persisted above 10(4) RNA copies/mL and either increased or declined during the later stages of disease; CD3+ CD4+ counts showed a steadily declining trend and the ratio of CD4 to CD8 decreased during late-stage disease. Moreover, antibodies against viral proteins were detected in the plasma and showed a significant increasing trend, while there were no apparently changes in the levels of IFN-gamma, IL-12, IL-2, IL-4, IL-10 and TNF-alpha. In conclusion, the characteristics of the SIV animal models in our study are similar to those of patients with AIDS. Therefore, the rhesus macaque SIVmac251 infection models can be applied for further studies into AIDS.
Animals
;
Antibodies, Viral
;
blood
;
CD4 Lymphocyte Count
;
CD4-Positive T-Lymphocytes
;
virology
;
Cytokines
;
genetics
;
immunology
;
Disease Models, Animal
;
HIV Infections
;
genetics
;
immunology
;
virology
;
HIV-1
;
physiology
;
Humans
;
Macaca mulatta
;
Male
;
Simian Acquired Immunodeficiency Syndrome
;
genetics
;
immunology
;
virology
;
Simian Immunodeficiency Virus
;
physiology
;
Viral Load