2.The effects of mitogen-activated protein kinase on EGF-induced expression of integrin α_5 in human RPE cells
Zhen, CHEN ; Yiqiao, XING ; Changzheng CHEN
Chinese Ophthalmic Research 2010;28(1):62-65
Background Human retinal pigment epithelial(RPE) cells play a critical role in the pathogenesis of proliferative vitreoretinopathy(PVR) and other related ocular diseases.Research demonstrated that epidermal growth factor(EGF) stimulates activation of RPE cells and therefore causes PVR,and integrin α_5 mediates the adhesion of cells and EGF.Objective This study aims to investigate the role of mitogen-activated protein kinase(MAPK) in regulation of EGF on integrin α_5 expression in human RPE cells.MethodsHuman RPE cells strain(CRL2302) was cultured in DMEM/F12 with 10% calf serum and passaged in 0.25% trypsin.Cultured cells were divided into DMEM/F12control group,20μmol/L PD98059 +10 ng/mL EGF+DMEM/F12(PD98059) group and 10 ng/mL EGF+DMEM/F12(EGF) group.The expression of integrin α_5 protein in CRL2302 cells was detected by RT-PCR and calculated as α_5 mRNA/β-actin mRNA,and the expression of integrin α_5 mRNA in CRL2302 cells was detected evaluated by flow cytometry.The MAPK phosphorylation level in each group of human RPE cells was tested by Western blot.ResultsThe expression value of integrin α_5 mRNA was 0.93±0.06 in the control group,1.06±0.07 in PD98059 group and 1.97±0.09 in EGF group,showing a significant difference among the three groups(F=458.896,P<0.01).The fluorescence intensity of integrin α5 protein in CRL2302 cells after 24 hours was 1.94±0.22,4.56±0.25,2.39± 0.14 in three groups respectively with a significant difference(F=21.05,P<0.05).After 30 minutes of culture,Western blot result showed that the strongest phosphorylation levels of ERK1/2 activation in EGF group and the weakest phosphorylation levels of ERK1/2 activation in the control group;While that in PD98059 group was significantly stronger than control group and weaker than EGF group(F=143.14,P<0.01).ConclusionEGF stimulates activation of ERK1/2 pathway in human RPE cells and the expressions of integrin α_5 mRNA and protein in human RPE cells in vitro.
3.Rationality of the Preventive Use of Antibiotics in Ophthalmologic Patients with Type Ⅰ Incision:Analysis of 1273 Cases
Ying XING ; Jianhua LIANG ; Jiancun ZHEN
China Pharmacy 2007;0(32):-
OBJECTIVE:To analyze the rationality of the preventive use of antibiotics in Chinese ophthalmologic patients with type Ⅰ incision.METHODS:By means of retrospective study,a total of 1 273 ophthalmologic cases from 113 hospitals of 2006~2007 from National Monitoring Network of Ministry of Health for Clinical Application of Antibiotic with type Ⅰ incision involving preventive administration of antibiotics via different routes were analyzed.RESULTS:The local administration (eye drops and local injection) was rational on the whole,but the systemic administration (intravenous or oral administration) was irrational to some degree in which the drug grade was on the high side,the antibacterial spectrum was inappropriate,and the medication time was delayed.CONCLUSION:Preoperative local administration and rational use of dosage forms should be emphasized in the preventive use of antibiotics for patients undergoing ophthalmologic surgery.
4.SIMPLIFICATION URETEROSCOPE TREATMENT THE OBSTRUCT IN URETER (REPORT OF 768 CASES)
Xing ZHOU ; Chunxiao LIU ; Shaobo ZHEN
China Journal of Endoscopy 2001;7(1):10-11
Objective:To study and sum up the experience of diagnosis and treatment of ureter obstruct by Simplification ureteroscope.Methods:the patients of 750 cases for ureterolith underwent ureteroscopic lithotomy and pressune orbit lithotripsy by using caudal anesthesia in outpatient the 11 cases ureterostensis were treated by watery capus expand,ureteroscopic hard expand and ureterotectomy by using of self-made electrode.Then,transurethral electro incisions of ureterocles were carried out for 2 patients.Results:Successful rate of ureteroscopic lithotomy is 98.4%;12 patient of upper ureter stone received ESWL because of stone entering the renal pelvis,and 3 cases complicated ureteral perforation;11 cases ureterostensis and 7 cases ureterocele were cured.Conclusions:It is frist chosen for ureteroscope to ureteral stone,ureteral stricture,and ureteroceles with a diameter of ≤3cm,and the method have ideal cure effect and the patients will receive.
5.Integrin α5 expression induced by epidermal growth factor affect proliferation and migration of human retinal pigment epithelium cells
Zhen CHEN ; Yiqiao XING ; Changzheng CHEN
Chinese Journal of Ocular Fundus Diseases 2010;26(3):267-270
Objective To observe the effect of epidermal growth factor (EGF) on integrin α5 expression and its influence on human retinal pigment epithelium (RPE) cells. Methods Human RPE cells were treated in vitro with 0.1, 1.0, 10.0, 20.0 and 100.0 ng/ml of EGF, the mRNA and protein of integrin α5 was measured by reverse transcription polymerase chain reaction(RT-PCR)and flow cytometry. Human RPE cells were cultured under 4 conditions including DMEM/F12, DMEM/F12 + 10 ng/ml EGF, DMEM/F12 + 10 ng/ml EGF+ rabbit anti-human integrin α5 antibody (1: 100), DMEM/F12 + 10 ng/ml EGF+ rabbit anti-human vimentin antibody (1: 100), and their proliferation and migration were measured by methyl-thiazole tetrazolium(MTT)and Boyden chamber. Results The integrin α5 mRNA level of human RPE cells was not changed after 12 hours of EGF stimulation (F=0.618, P = 0. 687), however it was induced in a dose-dependent manner after 24 and 48 hours of EGF stimulation (F=465. 303, 212. 340; P= 0. 000, 0. 000). The protein level of integrinα5 was higher in 10 ng/ml EGF stimulation compared with the control group and 0. 1 ng/ml group (P<0. 01). MTT and Boyden chamber showed that the integrin α5 expression increased the proliferation and migration of human RPE cells. Conclusion EGF can induce integrin α5 expression, thus increase the proliferation and migration of human RPE cells.
7.Research of expression of aldehyde dehydrogenase 1 and breast cancer resistance protein and their correlation in breast cancer
Xing WANG ; Zhen JIANG ; Changgang SUN
China Oncology 2014;(3):175-181
Background and purpose:Resistance to antitumor agents is a major cause of treatment failure in patients with breast cancer. Research has shown that, tumor stem cell marker aldehyde dehydrogenase 1 (ALDH1) is related with some anticancer drugs (such as cyclophosphamide, cisplatin) resistance, and the content of ALDH1 in tumor cells after treatment is higher than that before treatment. Breast cancer resistance protein (BCRP) is not expressed in normal tissues, but high expressed in breast cancer of after treatment, it may be associated with the mechanism of tumor drug resistance. This study was to investigate the correlation between expression and the relationship between these two kinds of protein ALDH1, BCRP and clinical pathological characteristics. Methods:Immunohistochemistry was performed to detect the expression of ALDH1 and BCRP in breast inifltrating ductal carcinoma tissues, and whether there is a correlation between and explore their relationship with clinical pathological features and their expression. Results:The expression of ALDH1 protein and BCRP protein in breast cancer and paracarcinoma breast tissues has signiifcant difference(χ2=14.685, P=0.000;χ2=12.243, P=0.000).The expression of ALDH1 with patients age, pathologic stage, axillary lymph node metastasis, histological grading, ER, PR, and HER-2 state were not relevant(P>0.05). HER-2, BCRP protein, expression was higher in cancer tissue (χ2=5.289, P=0.021). There were no relevant with the expression of BCRP with patients age, pathologic stage, axillary lymph node metastasis and histological grading, estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor (HER-2) (P>0.05). Conclusion:ALDH1 proteins may be an independent factor compared with occur drug resistant protein, and participate breast cancer drug resistance in the chemotherapy and tumor invasion and metastasis of malignant biological behavior.
8.Changes of visual development in critical period produced by monocular chronic atropinization in rats
Ya-Zhen, WU ; Yong-Xin, XING ; Hong, YAN
International Eye Science 2015;(4):592-595
AIM:To investigate the changes of visual development produced by monocular atropinization in rats.
METHODS: Twenty normal SD rats were randomly divided into two groups: control group ( n = 10 ) and atropinization ( experimental) group ( n=10 ) . All the left eyes were selected as the experimental eyes, and the right eyes served as the normal eyes. The left eyes in atropinization group was produced by 1% atropine, 3 times a day and the right eyes in control group was treated with normal saline, 3 times a day. The flash visual evoked potentials ( F-VEP ) and retinoscopy refraction of the rats'both eyes were detected at five time points:0, 7, 14, 21, and 28d after atropinization, respectively. After 28d, six rats were randomly selected from both groups and each group had three rats. The expression of the c- fos mRNA was observed in both visual cortexes. Another six rats were chosen for the same test after 2d dark environment with 2h light later. The expression of c-fos mRNA was detected again.
RESULTS: After 14d anisometropia was observed in experimental group, the difference was 3. 9D ( P<
0.0 5 ) , F-VEP P1 wave of the rats left in experimental group was reached to 88. 9±1. 889ms at 21d, there was statistical difference compared with the right eye ( P<0.05). After 28d, c-fos mRNA expression in the left visual cortex of rats in the experimental group was higher than that of the right side, but there was no significant difference. But when underwent 2h light stimulation after in the darkroom 2d, the c-fos mRNA expression in in the left visual cortex of rats in the experimental group was 5 times higher than that of the right side, there was a statistically significant difference (P<0. 05).
CONCLUSION: In the critical period of visual development, monocular chronic atropine in rats can form anisometropia, may delay the transmission of the optic nerve, hinder the normal development of the visual cortex. Monocular atropinization in rats can be used as the model of anisometropia.
9.Activation of Rho Kinase in Lung Tissue of Hypoxic Pulmonary Hypertension in Rats in Different Stages and Its Significance
xing-zhen, SUN ; xiang-yang, TIAN ; da-wei, WANG
Journal of Applied Clinical Pediatrics 1986;0(01):-
Objective To study the expression of Rho kinase and its functional activation in lung tissue from hypoxic pulmonary hypertension(HPH) rat model,and the effects of fasudil on HPH.Methods Seventy-two male Spraque-Dawley rats were randomly divided into the control group,hypoxic model group,and fasudil-intervention group[group with hypoxia and fasudil for 15 mg/(kg?d)],respectively.Mean pulmonary arterial pressure(mPAP) and right ventricle hypertrophy index(RVHI) were measured.Expression of Rho kinase mRNA and protein were examined by reverse transcriptase-polymerase chain reaction(RT-PCR) and Western blot,respectively.The phosphorylation of binding subunit of myosin phosphatase(MBS)-a substrate of Rho kinase was detected by Western blot and defined,as the mark of functional activation of the kinase.Results The expression of Rho kinase mRNA in hypoxic model group was markedly upregulated even before the onset of the third day after the experiment(HPH),and it was much lower in rats of fasudil group than that of hypoxic model group.The phosphorylation of MBS was significantly higher in hypoxic model group than that in control group,and it was positively correlated with the mPAP and RVHI(all P
10.Correlation between Single Nucleotide of Polymorphisms of -238,-308 G/A in Tumour Necrosis Factor-? Gene Promoter and Juvenile Idiopathic Arthritis
xing-gang, ZHEN ; shao, PENG ; yu-ju, CAO
Journal of Applied Clinical Pediatrics 2004;0(09):-
Objective To study the correlation between single nucleotide polymorphisms of -238,-308 G/A in promoter region of tumor necrosis factor -?(TNF-?) gene and the type of juvenile idiopathic arthritis (JIA).Methods Clinical data and blood preparation of 127 children with JIA and 106 healthy children were evaluated.Subgroups of JIA were defined according to the Edmonton criteria.The -238 G/A and -308 G/A polymorphisms in DNA analysis in this study were extracted from the whole blood.The restricted fragment length polymorphisms were determined in the cases of all JIA children and control group.Results 1.The TNF-?-238 G/A allele frequencies of JIA group and control group:allele frequency of JIA group was 92.9% and 7.1%,and the control group was 95.3% and 4.7%.The distribution of allele frequencies was no significantly different between JIA group and control group(?2=1.149 P=0.284).But there were significant difference between polyarticular JIA (RF negative) and control group(?2=7.621 P=0.006).2.The TNF-?-308 G/A allele frequencies of JIA group and control group:allele frequency of JIA group was 94.1% and 5.9%,the control group was 95.3% and 4.7%.The distributions of allele frequencies was no significantly different between JIA group and control group(?2=0.322 P=0.571).There were significantly difference between polyarticular JIA (RF negative) and control group (?2=7.621 P=0.006).Conclusions The TNF-?-238,-308 polymorphisms of A in the-238 and-308 TNF-? gene are important to the joint destruction of JIA.The study will be beneficial to provide indirect support to the application of anti-TNF drugs to the treatment of JIA.