1.Role of STAT3 in the regulation of autophagy in the glomerular mesangial cells of diabetic nephropathy
Yun CHEN ; Shuang YANG ; Lining WANG ; Dan SUN ; Xiuying WANG
Chinese Journal of Nephrology 2016;32(10):772-778
Objective To observe the changes of STAT3 signaling transduction pathway and autophagy activity in human glomerular mesangial cells cultured in high glucose, as well as the effect of STAT3 on autophagy, exploring whether SAT3 further influence extracellular matrix proteins type IV collagen secretion through the regulation of autophagy. Methods Culture human renal mesangial cells under different conditions, STAT3 pathway was inhibited with specific blocking agent S3I?201 and siRNA respectively. The experiment was divided into: (1) Control group: normal glucose concentration; (2) High glucose group: divided into 12 h, 24 h, 48 h, 72 h incubation group. (3) High glucose+S3I?201 group: pretreated cells with 30 μmol/L S3I?201 (Selleck S1155) for 1 h, then incubation with high glucose for another 24 hours. (4) High glucose+STAT3?siRNA group: siRNA transfection firstly, then incubation with high glucose for 24 hours. (5) High glucose+S3I?201+3?MA group: pretreated cells with 2 mmol/L 3?MA (Selleck S2767) and 30 μmol/L S3I?201 for 1 h, then incubation with high glucose for another 24 hours. Western blot was employed to detect the protein of STAT3, p?STAT3 and autophagy related protein LC3, p62 expressions. The changes of autophagosome quantity was observed with transmission electron microscope. The extracellular matrix protein collagen IV expression was measured with ELISA. Results Compared with the control group, glomerular mesangial cells cultured with high glucose for 24h, the expressions of STAT3 and p?STAT3 increased (P<0.01), while the expression of autophagy related proteins LC3II/LC3I decreased. The expression of p62 increased and the number of autophagosome reduced under transmission electron microscope, which all indicated the decrease of autophagy activity (P<0.05). Blocking STAT3 signaling pathway with S3I?201 and STAT3?siRNA respectively, compared with high glucose group, LC3II/LC3I was up?regulated and p62 was down?regulated, and the number of autophagosome was increased significantly, which all indicated the increase of autophagy activity (P<0.05). Extracellular matrix proteins collagen IV expression of cells cultured with high glucose was higher than the control group (P<0.05), and the application of S3I?201 blocking STAT3 pathway caused type IV collagen expression to decrease (P<0.05). The application of the autophagy inhibitor 3?MA could convert the result and lead to an increase of type IV collagen expression (P<0.01). Conclusions High glucose could active STAT3 signaling pathway of human renal mesangial cell and increase STAT3, p?STAT3 expression. High glucose could inhibit autophagy activity of human renal mesangial cells. Inhibition of STAT3 pathway activation may reduce the inhibitory effect of high glucose on autophagy of human renal mesangial cells. High glucose leads to an increase of type IV collagen secretion of human glomerular mesangial cells. The activation of STAT3 pathway may increase type IV collagen secretion through negative regulation of autophagy, which eventually leads to diabetic nephropathy.
2.Related Risk Factors between Diabetic Retinopathy and Diabetic Peripheral Neuropathy in Newly Diagnosed Type 2 Diabetes Mellitus
Min FANG ; Yun YU ; Zhenglong OUYANG ; Dan LIU ; Mei WANG
Journal of Sun Yat-sen University(Medical Sciences) 2017;38(2):315-320
[Objective]To analyze the clinical features and related risk factors in diabetic retinopathy(DR)and diabetic periph?eral neuropathy(DPN),the two micro-peripheral vascular diseases in newly diagnosed type 2 diabetes mellitus.[Methods]A retro?spective study of 211 cases of newly diagnosed type 2 diabetes mellitus inpatient from 2009 to 2012 ,and compare the two micro-peripheral vascular complications.[Results]The morbidity of DPN was(33.6%),which was higher than that of DR,some cases of DR co-existed with DPN,DR was related with DPN(r=0.158,P=0.020). Age and systolic blood pressure were the common risk factors in DR and DPN by single factor analysis.[Conclusion]It should be paid attention to the screening of both DR and DPN in each age group in newly diagnosed type 2 diabetes mellitus. Except for the control of blood glucose ,the control of the systolic blood pressure is important in the prevention and treatment in the two micro-peripheral vascular diseases.
4.Influence of different length corneal limbal incisions on the anterior and posterior corneal astigmatism after phacoemulsification
Yong-mou, ZHOU ; Ping-jun, CHANG ; Dan-dan, WANG ; Yun-e, ZHAO
Chinese Journal of Experimental Ophthalmology 2012;30(6):543-547
Background The different incisions in phacoemulsification,including the length,location and shape etc.,can cause surgery-induced astigmatism ( SIA ).But the SIA caused by 2.2 mm,3.0 mm corneal limbal incision after phacoemulsification,especially the change of posterior corneal surface astigmatism is still rarely reported. Objective This study was to investigate the anterior,posterior and total corneal SIA and compare their differences between phacoemulsification and foldable intraocular lens (IOL) implantation with 2.2 mm and 3.0 mm corneal limbal incisions. Methods Seventy-one eyes of 47 cases were randomly divided into two groups with matched age,visual acuity and astigmatism degree.Phacoemulsification and IOL implantation with 2.2 mm incision at the steepest corneal meridian was performed on the patients of 2.2 mm incision group,and the same surgery was adopted with 3.0 mm incision as 3.0 mm incision group.Corneal curvature radius and central corneal thickness were measured by Pentacam at 1 day before surgery and 1 week,1 month and 3 months after surgery respectively.The anterior and posterior corneal surface SIAs were calculated according to the flat axis and steep axis of corneal curvature and the air and the cornea refractive index.Based on the anterior and posterior surface SIAs,the total corneal SIA was then calculated using the vector analysis method.Jaffe/Clayman vector method was used to calculate the anterior and posterior and total corneal SIAs in the different time points,and the differences were compared between the two groups.Oral informed consent was obtained from each subject prior to the trial. Results The mean anterior and posterior surface corneal SIAs appeared to be lower in 2.2 mm incision group compared with 3.0 mm incision group at postoperative 1 day,1 week,1 month and 3 months but were not significantly different among groups at various time points ( anterior SIA:P =0.290 ; posterior SIA:P =0.740 ; total SIA:0.434 ).The mean anterior corneal surface SIAs were significantly lower at the postoperative 3 months than those at postoperative 1 day,1 week in both groups(2.2 mm incision group:P=0.020,0.036;3.0 mm incision group:P=0.006,0.023 ).The posterior corneal surface SIAs were (0.70±0.43 ) D and (0.75 ±0.54 ) D at 1 day in 2.2 mm incision group and 3.0 mm inscision group,respectively,and significantly decreased posterior corneal surface SIAs were found in postoperative 1 week,1 month and 3 months compared with 1 day in both groups ( 2.2 mm incision group:all P =0.001 ; 3.0 mm incision group:P=0.028,0.044,0.032).The total corneal surface SIA showed significant differences between 1 day and 1 week,1 month,3 months after surgery ( 2.2 mm incision group:P =0.015,0.002,0.002 ; 3.0 mm incision group:P =0.049,0.007,0.016 ). Conclusions There are no significant differences in the anterior,posterior and total corneal surface SIAs between 2.2 mm and 3.0 mm incisions after phacoemulsification with IOL implantation.The SIA is gradually reduced with the prolongation of postoperative time.
5.Total flavonoid from Glycyrrhizae Radix et Rhizoma and its ingredient isoliquiritigenin regulation M2 phenotype polarization of macrophages.
Yuan-lai WANG ; Xi TAN ; Xiao-lu YANG ; Xiao-yun LI ; Ka BIAN ; Dan-dan ZHANG
China Journal of Chinese Materia Medica 2015;40(22):4475-4481
To study the mechanisms of total flavonoid from Glycyrrhizae Radix et Rhizoma (TFGR) and its ingredient isoliquiritigenin (ISL) on their regulation of M2 phenotype polarization of macrophages. IL-4 (60 μg x L(-1)) induced RAW264.7 cells for 6 h to establish the M2 macrophage model. TFGR and ISL restrained breast cancer cells migration with the aid of M2 macrophages in vitro. TFGR and ISL inhibited gene and protein expression of Arg-1, up-regulated gene of HO-1 and protein expression of iNOS, enhanced the expression of microRNA 155 and its target gene SHIP1, meanwhile down-regulated.the phosphorylation of STAT3 and STAT6. So TFGR and ISL were the bioactive fraction and ingredient in Glycyrrhizae Radix et Rhizoma to reverse M2 phenotype macrophages polarization. TFGR and ISL inhibited the promotion of M2 macrophages to breast cancer cells migration in vitro, STAT signal pathways and miR155 were partly involved.
Animals
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Cell Line, Tumor
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Cell Movement
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drug effects
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Cell Polarity
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drug effects
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Chalcones
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pharmacology
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Flavonoids
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pharmacology
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Glycyrrhiza
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chemistry
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Interleukin-4
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genetics
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metabolism
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Macrophages
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cytology
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drug effects
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metabolism
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Mice
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RAW 264.7 Cells
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Rhizome
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chemistry
6.18F-FDG PET/CT in diagnosis of malignant pericardial lesions
Dan, SHAO ; Shu-xia, WANG ; Chang-hong, LIANG ; Si-yun, WANG ; Wei-ping, XU
Chinese Journal of Nuclear Medicine 2010;30(5):333-335
Objective To assess the value of integrated 18 F-fluorodeoxyglucose (FDG) PET/CT in differentiation of malignant and benign pericardial effusion. Methods 18F-FDG PET/CT were performed in 23 patients with pericardial effusion. The detected soft tissue tumor or nodulous lession in pericardium or the thickened pericardium, with the maximum standardized uptake value( SUVmax ) ≥2.5, was defined as PET/CT-positive. The invaded lession in pericardium with SUVmax ≥2.5 was also as the positive. The difference of SUVmax of benign and malignant lesions was analyzed with two-independent-sample test of nonparametric tests. The final diagnosis was confirmed by biopsy or post-operative pathology. Results The diagnosis were confirmed with 14 malignant and 9 benign lesions. The median of SUVmax was 6.0 in malignancy group and 2.2 in benign group (z= -3. 279, P =0.001 ). According to the pathology results, there were one false negative case and two false positive cases with PET/CT imaging interpretation. The sensitivity, specificity,accuracy, positive predictive value ( PPV ) and negative predictive value ( NPV ) of 18 F-FDG PET/CT in diagnosis of benignity or malignance of pericardium effusion were 92.9% ( 13/14), 7/9, 87.0% (20/23),86.7% (13/15) and 7/8, respectively. Conclusion For the patients with pericardium effusion 18F-FDG PET/CT may be a helpful modality for malignancy differentiation
7.Impact of psychological intervention to compliance and quality of life for cancer patients and their primary caregivers
Li JIA ; Dan WANG ; Lin JIA ; Jianjun HAN ; Yun WANG ; Fei GAO ; Mi CHEN ; Han SU
Cancer Research and Clinic 2012;(12):828-830
Objective To investigate the impact of compliance and quality of life of psychological intervention for cancer patients and their primary caregivers,as well as the correlation between the psychological issues of patients and their primary caregivers.Methods The enrolled patients were randomly divided into intervention group and control group.The patients in intervention group were given to standardize anti-tumor therapy,while the patients and their primary caregivers were given psychological intervention once a week.The patients in the control group only received standard anti-tumor therapy.By TDL determination of quality of life,anxiety rating scale (SAS) and self-rating depression scale (SDS),in front of the psychological intervention after 8 weeks of intervention,the two groups of patients and their primary caregivers were questionnaired,and recorded the completion of the treatment plan.By SPSS 12.0 software,the statistics were completed.Results 51 cases in intervention group and 38 cases in control group were able to complete the number of people expected to treat there was a statistically significant (P < 0.05).TDL determination and quality of life scores in intervention group patients and their primary caregivers were significantly higher (P < 0.05).SAS and SDS score in intervention group patients and their primary caregivers were significantly lower than control group (P < 0.05).Conclusion The effective psychological intervention to cancer patients and their primary caregivers during the treatment of patients could improve the compliance of cancer patients,the quality of life of cancer patients and their primary caregivers.The psychological problems between the patients and primary caregivers are positive correlation.
8.Effects of Bcl-2 associated with athanogene-1 gene silencing on heat shock protein 70 expression and human neuroblastoma cell apoptosis induced by hypoxia
Yankun SONG ; Zhi LI ; Fengtao WANG ; Haiyan LIU ; Yan QU ; Yun WANG ; Chunyu XIE ; Dan HU
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2017;24(1):68-72
Objective To investigate the protective effects of Bcl-2 associated with athanogene-1 (BAG-1) gene on human neuroblastoma cells (SH-SY5Y) injury induced by hypoxia/reoxygenation,and its influence on heat shock protein 70 (HSP70) expression.Methods The SH-SYSY cells at logarithmic growth phase were collected.Lenti virus mediated RNA interference (RNAi) technology was used to suppress the BAG-1 expression.The cells screened out can be divided into four groups:the cell control group with no lentivirus infection,lentivirus control group (containing only fluorescein protein lentivirus infection),BAG-1 siRNA group (BAG-1 siRNA silencing group),including BAG-1 siRNA-α group and BAG-1 siRNA-β group with lentivirus containing fluorescein protein (GFP) but at different BAG-1 siRNA target sites of silencing.Western Blot was used to detect the protein expression of BAG-1 and HSP70 in target cells after infectious recombination lentivirus for 72 hours;the Cell Counting Kit-8 (CCK-8) was used to detect the activity of four different group cells after hypoxia;the flow cytometry was used to detect the cell apoptosis;the HSP70 mRNA transcription level were detected by real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-qPCR) respectively in each group.Results After lentiviral infection for 72 hours,the Western Blot results showed that in the two BAG-1 siRNA silencing groups,the interference effect of BAG-1 siRNA-β group was superior to that of BAG-1 siRNA-α group.The cell viability of each group showed an increase followed by a decrease with the prolongation of hypoxia time,and reaching the peak at 8 hours.After hypoxia for 8 hours being given,the cell viability in BAG-1 siRNA-β group was significantly lower than that of the cell control group,lentivirus control group and BAG-1 siRNA-α group (A value:0.59 ±0.09 vs.0.94±0.12,0.90± 0.11,0.91± 0.14,P < 0.01);the cell apoptosis rate was obviously higher in BAG-1 siRNA-β than that in the above three groups [(34.63 ± 3.46)% vs.(14.83 ± 3.75)%,(19.93 ± 6.49)%,(16.40± 1.18)%,all P < 0.01].There were no statistically significant differences in the HSP70 protein level and mRNA transcription level between BAG-1 siRNA-3 grroup,and the cell control group,lentivirus control group and BAG-1 siRNA-α group respectively (all P > 0.05).Conclusion BAG-1 gene can play a role in protection of hypoxia nerve cells,reduce the apoptosis,and its protective effect can be independent of HSP70 gene.
9.Effects of lentivirus-mediated heat shock protein 70 gene on endoplasmic reticulum calcium homeostasis and calcium channels of PC12 cells induced by ischemia and hypoxia
Yuan LIU ; Chun GUAN ; Lulu GUO ; Qingshu LI ; Yun WANG ; Chunyu XIE ; Dan HU ; Yan QU
Chinese Critical Care Medicine 2016;28(3):205-210
Objective To investigate the effects of lentivirus-mediated heat shock protein 70 (HSP70) gene on calcium homeostasis and calcium channels of PC12 cells induced by ischemic and hypoxia and its mechanisms. Methods PC12 cells at logarithmic phase were collected, and they were divided into recombined lentiviral infection group [infected by lentivirus containing HSP70 and green fluorescent protein (GFP) fluorescin gene], lentivirus control group (infected by lentivirus containing GFP without HSP70 gene) and non-infection group. PC12 cells were subjected ischemia/hypoxia for 4, 8, 12, 24 hours, and the cell activity was determined by methylthiazolyl tetrazolium (MTT) assay test inorder to determine the best time for ischemia/hypoxia. The mRNA expressions of HSP70, muscle/endoplasmic reticulum Ca2+-ATP isoforms (SERCA2a, SERCA2b), ryanodine receptor 2 (RyR2), and inositol 1,4,5-triphosphate receptor 1 (IP3R1) were determined by real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR), and the protein expressions of HSP70, SERCA, and IP3R were determined by Western Blot at 8 hours after ischemic/hypoxia. Flow cytometry was used to determine the levels of intracellular reactive oxygen (ROS) and intracellular Ca2+ ([Ca2+]i). Results With the prolongation of time of ischemia/hypoxia, the cell viability in all groups showed an increase followed by a weakening, and peaked at 8 hours. The cell viability at 8 hours in lentiviral infection group was significantly higher than that of the non-infection group and lentivirus control group [A value (×10-2): 20.3±2.2 vs. 14.1±2.1, 15.0±1.6, both P < 0.01], the mRNA and protein expressions of HSP70 and SERCA in lentiviral infection group were significantly increased [HSP70 mRNA (2-ΔΔCt ): 0.785±0.018 vs. 0.428±0.019, 0.423±0.023; HSP70 protein (gray value): 2.72±0.20 vs. 1.56±0.36, 1.63±0.41; SERCA2a mRNA (2-ΔΔCt ): 0.971±0.037 vs. 0.367±0.014, 0.347±0.012; SERCA2b mRNA (2-ΔΔCt ): 8.869±0.162 vs. 3.015±0.091, 2.941±0.091; SERCA protein (gray value): 2.84±0.18 vs. 1.48±0.26, 1.52±0.29], and IP3R2 mRNA and protein expressions were significantly declined [IP3R2 mRNA (2-ΔΔCt ): 0.183±0.020 vs. 0.439±0.020, 0.433±0.040; IP3R2 protein (gray value): 1.15±0.12 vs. 1.91±0.20, 1.83±0.19], with statistically significant differences (all P < 0.01); no significant difference in RyR mRNA was found [2-ΔΔCt (×10-3): 1.97±0.63 vs. 2.02±0.22, 2.01±0.09, both P > 0.05]; the relative fluorescence intensity of ROS and [Ca2+]i in lentiviral infection group was significantly reduced (ROS: 30.54±1.23 vs. 58.03±1.97, 57.72±2.35; [Ca2+]i: 34.50±2.05 vs. 48.20±3.02, 46.80±2.75, all P < 0.01]. Conclusion Exogenous HSP70 can maintain calcium homeostasis in the endoplasmic reticulum of PC12 cells, affect the Ca2+ channel protein regulated by calcium channel IP3R and calcium pump SERCA, which may cause hypoxia/ischemia intracellular injury.
10.Clinical Analysis of Infantile Cytomegalovirus Hepatitis with Cholestasis
qing-wen, SHAN ; lin-lin, WANG ; xiang, YUN ; dan-mei, LIANG
Journal of Applied Clinical Pediatrics 2003;0(10):-
Objective To investigate the clinical features of infantile cytomegalovirus(CMV) hepatitis with cholestasis.Methods The clinical features of infantile CMV hepatitis with cholestasis were retrospected in 48 cases and the liver function were detected before and after therapy in 23 cases.Results Forty-eight cases of infantile cytomegalovirus hepatitis with cholestasis had different degrees of jaundice, hepatosplenomegaly and abnormal liver functions. After therapy of 23 cases, the clinical symptoms and serum biochemical parameters such as bilirubin, alanine aminotransferase were improved , but serum parameters of aspartate aminotransferase ,alkaline phospholipids and ?-glutamyltransferase had no improvement.Conclusion The theraphy of CMV hepatitis with cholestasis shall be individual.