1.Quantitative detection and correlation analysis of Txnip,Trx in aqueous humor of cortical cataract patients with type 2 diabetes
Lingfeng JIANG ; Dong SHI ; Qichang YAN
Recent Advances in Ophthalmology 2017;37(8):736-738,742
Objective To investigate the level and correlation between Thioredoxin (Trx) and Thioredoxin interacting protein (Txnip) in aqueous humor of cortical cataract patients with type 2 diabetes.Methods The Txnip and Trx levels were determined by Elisa in aqueous humor of 60 cataract patients with type 2 diabetes,which were divided into group A (control group,HbA1c <5.5%),group B (5.5% ≤HbA1c <6.5%) and group C (HbA1c>6.5%),and each group contains 20 patients.Results The level of Txnip in group B was higher than that in group A,while that in group C was the highest,there were statistical differences (all P < 0.05).Compared with group A,the level of Trx in group B was increased,while that in group C was the lowest,there were statistical differences (all P < 0.05).The level of Trx was positive correlated with Txnip in group A (r =0.810,P =0.000),but negative correlated with Txnip in group C (r =-0.809,P =0.000) and degree of cataract in group C (r =-0.727,P =0.001).Logistic regression analysis revealed that Txnip and HbAl c were risk factors of cortical cataract.Conclusion Trx and Txnip may play an important role in the development of diabetic cataract.
2.Identification of the binding region for interaction of optineurin with neural retina leucine
Chunxia, WANG ; Hongming, DONG ; Yu, ZHAO ; Qichang, YAN ; Jinsong, ZHANG
Chinese Journal of Experimental Ophthalmology 2014;32(8):677-681
Background Gene encoding optineurin (OPTN) is a causative gene for glaucoma and amyotrophic lateral sclerosis,with a more expression in retina.Our previous study isolated OPTN-interacting proteins and identified that the gene encode the basic leucine zipper (bZIP) transcription factor neural retina leucine (NRL) zipper,a causative gene for retinitis pigmentosa,and further study demonstrated the interaction between OPTN and NRL proteins in nuclei of cultured HeLaS3 cells.Objective This study was to determine the protein binding site of OPTN necessary for NRL binding.Methods A deletion series of OPTN-expression plasmids were constructed and co-expressed with hemagglutinin (HA)-tagged NRL in HeLaS3 cells,respectively.The cytoplasmic and nuclear fractions were used to perform co-immunoprecipitate (CoIP) and Western blot with anti-tag antibodies.Results In the nuclear fractions of cells transfected with the del1 st,del2nd or del3rd plasmid,a band of coimmunoprecipitated HA-labelled NRL (HA-NRL) was detected.However,the del4th plasmid did not produce a band.The NRL band was not found in cytoplasmic fractions from transfected cells with any of the deletion plasmids or with the whole-length OPTN plasmid.Conclusions The protein binding site of OPTN necessary for NRL binding is determined.This result demonstrates the binding of Flag-OPTN and HA-NRL in HeLaS3 cells.A series of partial-deletion OPTN plasmids demonstrated that the tail region (423-577 amino acids) of OPTN was necessary for binding with NRL.
3.SUMO Expression and Regulation in Oxidative Stress in Cultured Human Lens Epithelial Cells
Xiao HAN ; Xinling WANG ; Di WU ; Jinsong ZHANG ; Qichang YAN
Journal of China Medical University 2015;(3):193-198
Objective To observe the expression of small ubiquitin?related modifiers(SUMO)protein in normal cultured human lens epithelial cells(SRA01/04)and discuss regulation effects of SUMO protein on oxidative stress induced by high glucose. Methods The expression and local?ization of SUMO 1,2/3,4 was detected in normal cultured SRA01/04 cells through immunocytochemistry. The mRNA expression levels of SUMO 1?4 were examined by RT?PCR after the SRA01/04 cells treated with high glucose media at different concentrations and time points. Samples were grouped by medium concentrations(glucoses 5.5 mmol/L,12.5 mmol/L,25 mmol/L,50 mmol/L respectively for 24 h)and by treatment time(0 h, 6 h,12 h and 24 h respectively). After highly efficient transfection of GFP?SUMO2 into SRA01/04 cells,the survival and apoptotic rates of transfect?ed and un?transfected cells treated with high glucose was detected by CCK8 method and AV/PI double staining flow cytometry. Results The immu?nocytochemistry results showed that SUMO1,2/3,4 proteins were mainly located in the nucleus of SRA01/04 cells and part of SUMO2/3 was in the cytoplasm. RT?PCR results showed that compared with the low?glucose group,the mRNA expression of SUMO1?4 was increased along the increas?ing glucose concentration in the high?glucose group(P<0.05). Compared with 0 h,the mRNA expression of SUMO1?4 was enhanced at 6 h,12 h and 24 h(P<0.05)in the high?glucose group treated at 50 mmol/L concentration. Compared with the un?transfected cells,the survival rate was in?creased and the apoptotic rate was decreased in GFP?SUMO2 transfected cells in oxidative stress induced by high glucose(P<0.05). Conclusion SUMO protein was positively expressed in SRA01/04 cells and the expression of SUMO mRNA was affected by oxidative stress induced by high glu?cose.
4.Expression and clinical significance of Kif2a in breast carcinoma by the application of tissue microarray
Yan ZHU ; Qichang YANG ; Hongbin LIU ; Xiaojuan ZHANG ; Yi SHEN ; Yongqiang SUN
The Journal of Practical Medicine 2016;32(16):2663-2666
Objective To study the expression and clinical significance of Kif2a in breast carcinoma by the application of tissue microarray. Methods Tissue microarray and immunohistochemistry method (S-P) were used to examine the expression of Kif2a in 102 cases of breast carcinoma tissue and adjacent tissue. The prognosis of 62 cases followed up were studied by both univariate and multivariate analysis. Results The expression of Kif2a has significant difference between breast carcinoma tissue and adjacent tissue. Kif2a expression were associated with lymph nodal involvement and HER2 expression (P < 0.05). In the 62 cases with followed-up data, univariante analysis revealed that the expression of Kif2a, lymph nodal involvement and HER2 expression were correlated with the survival rate. Cox stepwise proportional hazards analysis showed that they were independently prognostic factors. Conclusions Kif2a expression in breast carcinoma were associated with lymph nodal involvement and HER2 expression (P<0.05). It is meaningful to examine Kif2a expression for prognosis of the breast carcinoma.
5.Prenatal diagnosis and outcome of congenital cardiac diventiculum
Shi ZENG ; Qichang ZHOU ; Jiawei ZHOU ; Ming ZHANG ; Qinghai PENG ; Xing YAN
Chinese Journal of Ultrasonography 2015;24(1):16-18
Objective To investigate the clinical outcome and prenatal diagnosis feasibility of fetal congenital cardiac diventiculum.Methods The data of 12 fetuses with congenital cardiac diventiculum were retrospectively reviewed.The prenatal and postnatal medical records,including the characteristics of diverticulum,presence of abnormalities,karyotype and the outcomes of each pregnancy were collected.Results The overall incidence of cardiac diventiculum was 0.03% (12/40 564) and the rate of incidence between left and right ventricle was 2 ∶ 1.Mean size of diventiculum was (69.75 ± 28.73)mm2,mean diameter of the diventiculum neck was (3.58 ± 0.80) mm and mean thickness of diventiculum wall was (1.54± 0.29)mm.Seven cases (58.4%) as an isolated malformation,5 cases (41.6%) combined with cardiac defect and extracardiac abnormalities,2 cases (16.7 %) with chromosomal abnormalities.Five cases underwent termination of pregnancy,1 case died in uterus and 6 cases were born live.The mean follow-up periods was (62.33±-36.52)month.Of the 6 follow-up cases,4 cases (66.7%) remained asymptomatic,one case underwent drug therapy because of arrhythmia and one case combined with VSD underwent operation.Conclusions Echocardiography could be an useful tool to demonstrate and monitor congenital cardiac diventiculum prenatally and postnatally.The outcome of cardiac diventiculum depends on the size,progression,and the combined abnormalities and complications.
6.Mechanism of Eaf2 Gene Regulating microRNA in Inhibiting the Genesis of Cataract
Yu QIN ; Jiangyue ZHAO ; Wenting LUO ; Jing LI ; Xinwei WU ; Jia LIU ; Qichang YAN ; Jinsong ZHANG
Journal of China Medical University 2015;(3):199-202
Objective To explore the expression of microRNA in Eaf2 knockout mice and the effect of Eaf2 on the apoptosis of human lens epithe?lial cells and the expression of microRNA in lens. Methods pEGFP?C1?Eaf2 was transfected into SRA01/04 cells using Lipofectamine 2000 to over express Eaf2 gene,and then the flow cytometry was used to detect cell apoptosis rate. And real time q?PCR was used to measure the expression of microRNA both in human lens epithelial cells and Eaf2 knockout mice. Results Compared with controls,the apoptosis rate of cells transfected with pEGFP?C1?Eaf2 was reduced,the expression of miR?125b and let?7a was significantly increased and miR?204 was decreased in cells transfect?ed with pEGFP?C1?Eaf2. Compared with controls,the expression of miR?125b and let?7a was lower and miR?204 was higher in Eaf2 knockout mice. Each result was statistically significant(P<0.01). Conclusion Eaf2 might inhibit apoptosis of human lens epithelial cells via regulating the expression of microRNA. Eaf2 may have a protective effect for the lens in the genesis of cataract.
7.Expression and clinicopathological significance of ADAM17 mRNA in esophageal squamous cell carcinoma
Hongbin LIU ; Qichang YANG ; Yi SHEN ; Yan ZHU ; Xiaojuan ZHANG ; Hao CHEN
Cancer Research and Clinic 2012;24(7):444-446,450
ObjectiveTo investigate the clinicopathologic significance of a disintegrin and metalloproteinase 17(ADAM 17) mRNA in esophageal squamous cell carcinoma (ESCC),and to evaluate its relation to clinicopathological features. MethodsThe expression of ADAM17 mRNA in 50 ESCC and 50 normal esophageal tissues were detected by RT-PCR. ResultsThe mRNA expression of ADAM17 in 50 ESCC and 50 normal esophageal tissues were 0.937±0.241 and 0.225±0.077,respectively.The expression of ADAM17 mRNA in the ESCC was much higher than in normal esophageal tissues (t=-19.899,P<0.01).The expression of ADAM 17 mRNA was positively correlated with lymph node metastasis(t=-4.703,P<0.01 ) and TNM staging (t=-2.652,P<0.05).There were no correlation between the expression of ADAM17 mRNA and sex,age and histological grade (t=0.299,-0.907,-3.163,all P>0.05).ConclusionADAM17 mRNA was higherly express in ESCC than in normal esophageal tissues and may play an important role in the development,invasion and metastasis of ESCC.It may be used as a prognostic factor.
8.Effect of Yirui Capsules on Serum Inflammatory Cytokines and Hepatic Alpha-7 Nicotinic Acetylcholine Receptor Expression in Hyperlipidemia Rats
Haisong ZHOU ; Lang YI ; Wenzhi LI ; Qichang LUO ; Qing WANG ; Yan DONG ; Peixun WANG
Journal of Guangzhou University of Traditional Chinese Medicine 2015;32(6):1047-1051,1146
Objective To study the relationship between the inflammatory cytokine levels and the expression of hepatic alpha-7 nicotinic acetylcholine receptor (α7 nAChR) in hyperlipidemia rats and to investigate the intervention effect of Yirui Capsules ( mainly composed of Radix Salviae Miltiorrhizae, Fructus Crataegi, Rhizoma Alismatis, etc). Methods Fifty male SD rats were randomly divided into blank control group ( fed with conventional diet, N=10) and high-fat diet group (fed with high-fat diet, N=40). After feeding for 14 d, the forty rats in high-fat diet group were further randomly divided into model control group, and low-, middle-, and high-dose Yirui Capsules groups (140, 280, 560 mg/kg, respectively), 10 rats in each group and the treatment lasting 30 days. And then, the serum levels of lipids were detected, C-reactive protein (CRP) and tumor necrosis factor (TNF) -α in the serum were detected by enzyme-linked immunosorbent assay ( ELISA) , and the expression of hepatic α7 nAChR was analyzed by immunohistochemical staining and reverse transcription-polymerase chain reaction (RT-PCR) . Results Yirui Capsules in different dosages could improve serum lipid levels in hyperlipidemia rats to various degrees. The contents of serum inflammatory cytokines CRP and TNF-α and the mRNA expression of hepatic α7nAChR were increased in the model control group as compared with those in the blank control group ( P<0.01). The contents of inflammatory cytokines CRP, TNF-αand the mRNA expression of hepatic α7nAChR were decreased in the three Yirui Capsules groups as compared with those in the model control group ( P<0.01). The results of immunohistochemical staining for α7nAChR expression were consistent with RT-PCR results. Conclusion The expression of hepatic α7nAChR is increased in hyperlipidemia rats. Yirui capsules are effective on decreasing inflammatory cytokine levels, improving lipid metabolism, and down-regulating α7nAChR expression.
9.Relationship between the co-expression of CD105 and cyclin D1 and lymph node metastasis and prognosis of esophageal squamous cell carcinoma
Hongbin LIU ; Junbo QIAN ; Qichang YANG ; Xinli TU ; Yan ZHU ; Yi SHEN
Chinese Journal of Digestive Surgery 2010;09(4):280-282
Objective To investigate the relationship between the co-expression of CD105 and cyclin D1 and lymph node metastasis and prognosis of esophageal squamous cell carcinoma ( ESCC ). Methods Eighty cases of ESCC tissue were collected at The First People's Hospital of Nantong. The expression of CD105 and cyclin D1 of ESCC was detected by immunohistochemistry. The relationship between the co-expression of CD105 and cyclin D1 and lymph node metastasis and prognosis of ESCC was analysed. Eighty normal esophageal tissues were selected as controls. All data were analysed by t test, chi-square test, and Pearson correlation analysis. Survival was analysed by the Kaplan-Meier survival curve. Microvessel density (MVD) was used to indicate the expression of CD105 in the form of -x±s. Results The expression of CD105 in ESCC tissues was higher (36±8) than that in normal esophageal tissues ( 11±3) (t =25. 129, P<0.05). The positive rate of cyclin D1 expression in ESCC tissues was 61% (49/80), which was significantly higher than 23% (18/80) in normal esophageal tissues ( x2 =4.972, P<0.05). The MVD value of 44 patients was ≤36 (LCD105), and nine of them had lymph node metastasis. The MVD value of the remaining 36 patient was > 36 ( HCD105 ), and 26 of them had lymph node metastasis. Twenty-eight patients with positive expression of cyclin D1 had lymph node metastasis, while seven patients with negative expression of cyclin D1 had lymph node metastasis. The results of Pearson correlation analysis revealed that a high expression of CD105 and a positive cyclin D1 expression were correlated with lymph node metastasis (x2 =21.562, 9.217, P<0.05). The survival times of 28 patients with positive cyclin D1 and HCD105,21 patients with positive cyclin D1 and LCD105, eight patients with negative cyclin D1 and HCD105, and 23 patients with negative cyclin D1 and LCD105 were (31±6) months, (47±7) months, (51±9) months and (61±5) months, respectively, with a significant difference among the four groups (F = 11.76, P < 0. 05 ).Conclusion Co-expression of CD105 and cyclin D1 may be used as a prognostic factor of ESCC.
10.MR imaging and dynamic contrast enhancement findings of choroidal hemangioma
Qinghua CHEN ; Zhenchang WANG ; Junfang XIAN ; Qichang TIAN ; Fei YAN ; Bentao YANG ; Zhonglin LIU
Chinese Journal of Radiology 2009;43(7):735-738
Objective To characterize the regular and the dynamic contrast enhancement MR imaging in choroidal hemangioma.Methods MR imaging findings of 30 cases (31eyes, 32 lesions) with choroidal hemangioma confirmed by follow-up results were retrospectively analyzed.Among them, postcontrasted T1-weighted imaging was performed in 30 patients and dynamic contrast enhancement scanning was performed in 26 cases.MRI findings and the time-intensity curve of dynamic contrast enhancement were analyzed.Results Among the 32 choroidal hemangiomas, 26 of them were at the temporal side of optic disc and 28 lesions were fusiform.Before enhancement, 23 lesions showed isointense T1-weighted signal and 31 lesions were isointense on T2-weighted imaging.All the lesions showed strong enhancement on postcontrast T1-weighted imaging, including 31 homogenously enhanced lesions and one heterogeneously enhanced lesion.Retinal detachments were found in 18 eyes.Fill-in sign were observed in 12 lesions during dynamic contrast enhancement.The time-intensity curve of dynamic contrast enhancement in 28 lesions suggested a pattern with rapid enhancement and slow washout, time to peak (91.00±25.27) s, slope ratio 3.03±1.13, the median of washout ratio 17.06%, enhancement ratio 2.87±0.79.Conclusion MRI showed a few features of the location, shape, signal characteristics, and enhancement pattern in choroidal hemangioma, which may contribut to diagnosis and treatment plan of this disease.