1.Regulation of miR-155 in H 2O 2-induced oxidative stress in lens epithelial cells via targeting SIRT1
Yantao NIU ; Li ZHANG ; Wenfang XIE
Chinese Journal of Experimental Ophthalmology 2022;40(5):404-413
Objective:To investigate the role of microRNA (miR)-155 in hydrogen peroxide (H 2O 2)-induced oxidative stress injury in lens epithelial cells (LECs) and its mechanism regulating silent information regulator factor related enzymes 1 (SIRT1). Methods:The HLE-B3 at the logarithmic growth phase was taken and cultured for 24 hours under different concentrations of H 2O 2 (0, 50, 100, 200, 400, 800 μmol/L), and the cell viability was detected by MTT assay to determine the optimal concentration of H 2O 2 for establishing an oxidative stress injury model.HLE-B3 cells were divided into 6 groups, untreated blank control group, model control group cultured with 100 μmol/L H 2O 2, miR-155 mimics group transfected with miR-155 mimics, miR-155 mimics negative control group transfected with miR-155 mimics negative control, miR-155 inhibitor group transfected with miR-155 inhibitor, and miR-155 inhibitor negative control group transfected with miR-155 inhibitor negative control.Transfected cells were cultured with 100 μmol/L H 2O 2.Cells in various groups were cultured for 24 hours, and cell morphology was observed under an inverted microscope.The relative expression of miR-155 and SIRT1 mRNA in cells was assayed by fluorescent quantitative PCR.Cell apoptosis rates were detected by flow cytometry.Reactive oxygen species (ROS) content was identified by 2', 7'-dichlorofluorescein diacetate (DCFH-DA) fluorescent probe method.Superoxide dismutase (SOD) activity and malondialdehyde (MDA) concentration were measured by ELISA method.The targeting of SIRT1 by miR-155 was tested by dual luciferase reporter gene system.Expressions of SIRT1, B-cell lymphoma/leukemia-2 gene (bcl-2), bcl-2 associated X protein (bax), cleaved-cysteine aspartase 3 (cleaved-Caspase-3) proteins were determined by Western blot. Results:With the increase of H 2O 2 concentration, the cell viability gradually decreased, and the differences in cell viability among different concentrations were statistically significant (all at P<0.05), and 100 μmol/L was selected as the experimental concentration.Cells in blank control group grew well adherently.The number of cells in model control group decreased, and the morphology of some surviving cells changed, and their boundaries were blurred.There were fewer cells in miR-155 mimics group than model control group, and the cell morphology changed.There were more cells in miR-155 inhibitor group than model control group, and the cells grew well.Compared with model control group, the relative expression level of miR-155, the apoptosis rate, ROS content, MDA concentration, as well as the relative expression levels of bax and cleaved-Caspase-3 proteins were increased, and the relative expression level of SIRT1 mRNA, the SOD activity, the relative expression of SIRT1 and bcl-2 proteins, as well as bcl-2/bax were decreased in miR-155 mimics group, and the differences were statistically significant (all at P<0.05). Compared with model control group, the relative expression of miR-155, the apoptosis rate, ROS content, MDA concentration, as well as the relative expression levels of bax and cleaved-Caspase-3 proteins were decreased, and the relative expression level of SIRT1 mRNA, SOD activity, the relative expression levels of SIRT1 and bcl-2 protein, as well as bcl-2/bax were significantly increased in miR-155 inhibitor group, and the differences were statistically significant (all at P<0.05). The relative luciferase activity of wild-type SIRT1 in cells transfected with miR-155 mimics was 0.41±0.07, which was significantly weaker than 1.00±0.11 in cells transfected with miR-155 mimics negative control, and the relative luciferase activity of wild-type SIRT1 in cells transfected with miR-155 inhibitor was 1.98±0.17, which was significantly higher than 1.00±0.12 in cells transfected with miR-155 inhibitor negative control, showing statistically significant differences ( t=7.838, 8.157; both at P<0.05). No obvious effect on the relative luciferase activity of mutant SIRT1 was found in transfected cells. Conclusions:miR-155 is involved in H 2O 2-induced oxidative damage of LECs, and its overexpression can target the expression of SIRT1 and play a role in cell injury.
2.Ultrasonic characteristic analysis of non-lactation mastitis
Jun QIU ; Qin CHEN ; Jinglai XIE ; Xing GAO ; Wenfang ZHANG
Journal of Chinese Physician 2021;23(11):1690-1692,1698
Objective:To analyze and summarize the characteristics of ultrasonographic images of non-lactating mastitis, improve the diagnostic accuracy, and guide the clinical treatment.Methods:The ultrasonic images of 46 patients with non lactation mastitis diagnosed by pathology in People′s Hospital of Heyuan from January 2017 to November 2020 were retrospectively analyzed. The ultrasonic images were compared with the pathological results, and the ultrasonic characteristics of non lactation mastitis were analyzed.Results:Among the 46 cases of non lactation mastitis, there were 24 cases of chronic mastitis, 15 cases of granulomatous lobular mastitis, 5 cases of plasma cell mastitis and 2 cases of lymphocytic lobulitis. The ultrasonographic features of non lactation mastitis were patchy hypoechoic mixed with anechoic areas with poor sound transmission. Granulomatous lobular mastitis was easy to be misdiagnosed as malignant lesions because of its disordered echo.Conclusions:Mastering the ultrasound characteristics of non-lactation mastitis can help to improve the accuracy of diagnosis of non-lactation mastitis.
3.Sensitivity and specificity of nucleic acid testing in close contacts of COVID-19 cases in Guangzhou
Wenting ZHANG ; Dan LIU ; Chaojun XIE ; Dong SHEN ; Zongqiu CHEN ; Zhihao LI ; Yuan LIU ; Xiru ZHANG ; Peiliang CHEN ; Wenfang ZHONG ; Pei YANG ; Qingmei HUANG ; Lei LUO ; Chen MAO
Chinese Journal of Epidemiology 2021;42(8):1347-1352
Objective:To analyze the sensitivity and specificity of SARS-CoV-2 nucleic acid testing in 20 348 close contacts of COVID-19 cases in different prevention and control stages in Guangzhou and to provide scientific evidence for optimizing epidemic response strategies.Methods:A total of 20 348 close contacts of COVID-19 cases in Guangzhou were traced between February 21 and September 22,2020. All the close contacts were tested for the nucleic acid of SARS-CoV-2. The sensitivity and specificity of nucleic acid testing and diagnosis in the different prevention and control stages were compared.Results:In 20 348 close contacts, 12 462 were males (61.24%), the median ( P 25, P 75) of age of them was 31.0 years (23.0,43.0), the median number ( P 25, P 75) of nucleic acid testing for them was 2.0 (1.0,3.0), and the median ( P 25, P 75) of their quarantine days was 12.0 (8.0,13.0) days, respectively. A total of 256 COVID-19 cases were confirmed in the close contacts after seven nucleic acid tests. In the 1 st, 2 nd, 3 rd and 7 th nucleic acid testing, the sensitivity and specificity were 69.14% and 99.99% (177 cases confirmed), 89.84% and 99.99% (230 cases confirmed), 97.27% and 99.99% (249 cases confirmed), and 100.00% and 99.98%, respectively. In the three stages of COVID-19 prevention and control in China: domestic case stage, imported case stage, and imported case associated local epidemic stage, the sensitivity of the 1 st nucleic acid testing was 70.68%, 68.00% and 67.35%, and the specificity was 99.98%, 100.00% and 100.00%, respectively. Conclusions:The sensitivity of nucleic acid testing in the close contacts at the different stages were consistent with slight decrease, which might be related to the increased proportion of asymptomatic infections in the late stage of epidemic prevention and control with COVID-19 in Guangzhou. It is suggested to give three nucleic acid tests to improve the sensitivity and reduce false negative risk.
4.Shear wave elastography in the evaluation of fibrosis degree in renal allograft
Daopeng YANG ; Yan WANG ; Bowen ZHUANG ; Xiaoer ZHANG ; Guangliang HUANG ; Wenfang CHEN ; Gang HUANG ; Xiaohua XIE
Chinese Journal of Ultrasonography 2020;29(10):875-880
Objective:To explore the value of shear wave elastography imaging(SWE) in the diagnosis of renal allograft fibrosis and analyze its advantages and limitations.Methods:The renal allograft of 61 patients who underwent renal allograft biopsy from June 2019 to April 2020 in the First Affiliated Hospital of Sun Yat-sen University were included in this study. According to the Banff classification, there were 51 patients with mild-degree fibration(interstitial fibrosis/tubular atrophy, IFTA 0-Ⅰ), and 29 patients with moderate or severe-degree fibration(IFTA Ⅱ-Ⅲ). Two-dimensional ultrasound, color Doppler flow imaging, SWE and kidney function test performed. All the results were compared between the two groups. Diagnostic performance was evaluated by receiver operating characteristic (ROC) curve using pathology as gold standard. The sensitivity, specificity, positive predictive value and negative predictive value were calculated for the diagnosis of moderate or severe-degree fibration.Results:Compared to mild-degree fibration group, creatinine( P<0.001), glomerular filtration rate( P<0.001), RI of arcuate arteries( P=0.022) and SWE value( P<0.001) significantly increased in the moderate or severe-degree fibration group. There were significant correlations between IFTA and creatinine ( r s=0.488, P<0.001), glomerular filtration rate ( r s=-0.452, P<0.001), RI of arcuate arteries( r s=0.228, P=0.042), SWE value( r s=0.584, P<0.001). Taking the cutoff value of SWE value deduced by ROC curve as 21.7 kPa, the area under ROC curve was 0.827. The sensitivity, specificity, positive predictive value and negative predictive value were 86.2%, 74.5%, 61.0% and 89.7%, respectively. Conclusions:There is a good correlation between the SWE value and the degree of fibrosis in the transplanted kidney. SWE can be used to distinguish mild from moderate or severe fibrosis of renal allograft, providing a potential noninvasive method for the assessment of kidney allograft fibration.
5.Diagnostic value of contrast-enhanced ultrasound combined with tumor markers in benign and malignant ovarian tumors
Yiyun XU ; Mang LI ; Yan CAI ; Yongjin PENG ; Yingying YUAN ; Wenfang WU ; Chongwen XIE
Journal of Chinese Physician 2019;21(8):1146-1148
Objective To explore the diagnostic value of contrast-enhanced ultrasound combined with tumor markers in benign and malignant ovarian tumors.Methods The clinical data of 100 patients with ovarian tumors treated in our hospital from May 2016 to May 2017 were analyzed retrospectively.The parameters of contrast-enhanced ultrasonography (CEUS) for benign and malignant ovarian tumors were analyzed.The levels of tumor markers were measured and the ultrasound imaging was analyzed.Analysis of the detection rate of benign and malignant ovarian tumors by contrast-enhanced ultrasound alone or in combination with tumor markers.Results Compared with benign ovarian tumors,ovarian malignant tumors have the characteristics of uneven enhancement,early onset and peak time of contrast enhancement,and higher rate of enhancement (P < 0.05);compared with benign ovarian tumors,serum CA125 and CA199 levels in patients with ovarian malignant tumors were higher,with significant difference (P < 0.05).The detection rates of contrast-enhanced ultrasound combined with tumor markers were higher than those of ultrasound or tumor markers alone,and were in accordance with pathological criteria.Conclusions The application of combined contrast-enhanced ultrasound and tumor markers in the diagnosis of ovarian benign and malignant tumors has a high detection rate,can significantly improve the accuracy of clinical detection,and provide a strong basis for treatment.
6.Study on Chemical Constituents of Ethanol Extract from Yao Medicine Cissampelopsis spelaeicola
Shuang LIANG ; Wenfang MA ; Kewen LIANG ; Lichun ZHAO ; Gang DENG ; Dongyang XIE
China Pharmacy 2019;30(20):2835-2839
OBJECTIVE: To study the chemical constituents of ethanol extract from Yao medicine Cissampelopsis spelaeicola. METHODS: The petroleum ether, ethyl acetate and n-butanol fraction from 75% ethanol extract of C. spelaeicola were isolated and purified by silica gel, SephadexLH-20 gel column and AB-8 macroporous resin column, etc. The structures of the compounds were analyzed and identified by physicochemical properties and spectral data (mass spectrometry, hydrogen spectrum, carbon spectrum). RESULTS & CONCLUSIONS: Twelve compounds were isolated and identified from 75% ethanol extract of C. spelaeicola. β-sitosterol(Ⅰ) and Stigmasterol(Ⅱ) were isolated from petroleum ether fraction; p-hydroxybenzoic acid(Ⅲ), β-daucossterol(Ⅳ), protocatechuic acid(Ⅴ), 6β-hydroxyeremophi-7(11)-en-12,8β-olide(Ⅵ), 10β-hydroxyeremophil-7(11)-en-8,12-olide(Ⅶ), 10β-hydroxyeremophi-7(11),8(9)-dien-8,12-olide(Ⅷ), Quercetin(Ⅸ), Hyperin(Ⅹ) and 4α-hydroxy- eudesman-11-ene(Ⅺ) were isolated from ethyl acetate fraction; quercetin-3-O-robinobioside(Ⅻ) was isolated from n-butanol fraction. Compounds Ⅰ-Ⅻ are isolated from C. spelaeicola for the first time. The study can lay material foundation for activity stady of C. spelaeicola.
7. Clinical and research progress of congenital adrenal hyperplasia with adrenal rest tumor
Lingli XIE ; Qing ZHANG ; Wenfang XIA
Chinese Journal of Endocrinology and Metabolism 2019;35(9):795-800
Adrenal rest tumor (ART) is a well-known complication in congenital adrenal hyperplasia (CAH), which may occur in all age groups, especially in adolescence and adulthood. Tumors that occur in the testicular known as testicularadrenal rest tumor (TART), in the ovarian and parovarian tissue known as ovarian adrenal rest tumor (OART). ART is one of the major causes of reproductive dysfunction in adult with CAH. Early prevention and treatment is the key to avoid irreversible damage to gonadal function. In the past, we paid more attention to TART in children, while ignored TART and OART in adults. This article will review the origin of tumor cells, pathogenesis, epidemiology, diagnosis, treatment and prognosis of CAH associated with ART, which aims to guide clinical work through the profound understanding of CAH with ART.
8.Curative effect of ozone hydrotherapy for pemphigus
Fuqiong JIANG ; Danqi DENG ; Xiaolan LI ; Wenfang WANG ; Hong XIE ; Yongzhuo WU ; Chunyan LUAN ; Binbin YANG
Journal of Central South University(Medical Sciences) 2018;43(2):152-156
Objective:To determine clinical curative effects of ozone therapy for pemphigus vulgaris.Methods:Ozone hydrotherapy was used as an aid treatment for 32 patients with pemphigus vulgaris.The hydropathic compression of potassium permanganate solution for 34 patients with pemphigus vulgaris served as a control.The main treatment for both groups were glucocorticoids and immune inhibitors.The lesions of patients,bacterial infection,usage of antibiotics,patient's satisfaction,and clinical curative effect were evaluated in the 2 groups.Results:There was no significant difference in the curative effect and the average length of staying at hospital between the 2 groups (P>0.05).But rate for the usage of antibiotics was significantly reduced in the group of ozone hydrotherapy (P=0.039).The patients were more satisfied in using ozone hydrotherapy than the potassium permanganate solution after 7-day therapy (P>0.05).Conclusion:Ozone hydrotherapy is a safe and effective aid method for pemphigus vulgaris.It can reduce the usage of antibiotics.
9.Research on screening carbapenemase-producing Enterobacteriaceae by modified Hodge test
Chunbao XIE ; Jiangrong LUO ; Liangmin CHUAN ; Daiwen XIAO ; Hua YU ; Yongchang YANG ; Wei JIANG ; Wenfang HUANG
International Journal of Laboratory Medicine 2017;38(15):2034-2035,2039
Objective To discuss the application value of modified Hodge test(MHT) for screening carbapenemase-producing Enterobacteriaceae.Methods The 24 Enterobacteriaceae reduced susceptibility to carbapenems were detected by MHT.At the same time,polymerase chain reaction(PCR) was used to detect carbapenemase genes of KPC,NDM,IMP,SIM and VIM.PCR products were sequenced and the results were compared with the sequences of Gen Bank database.Comprehensive analysis the application value of MHT and PCR to detect carbapenemase.Results Among these 24 strains,13 stains appeared to produce carbapenemase by MHT,5 positive strains were found to carry carbapenemase genes by PCR.By comparing with the sequences of Gen Bank database 1 strain were confirmed to KPC-2 and 4 strains were confirmed to IMP-4.We found that 4 strains of Enterobacteriaceae,detected carbapenemase by MHT and PCR at the same time.9 strains of MHT were positive,but we couldn′t detect the carbapenemase genes.1 strain of MHT was negative,but carbapenemase gene was found in the strain.Conclusion The value of MHT to screen carbapenemase-producing Enterobacteriaceae is necessary to further study.
10.Analysis on detection results of whole blood trace elements among 786 children aged 0-6 years old in Chengdu City
Min ZHONG ; Chunbao XIE ; Liangmin CHUAN ; Wenfang HUANG
International Journal of Laboratory Medicine 2016;37(16):2244-2246
Objective To know the contents and changes situation of trace elements among children aged 0-6 years old in Chengdu are‐a .Methods The BH5100T atomic absorption spectrometry was adopted to detect the levels of trace elements Cu ,Zn ,Ca ,Mg ,Fe in 786 children aged 0-6 years old undergoing the physical examination in our hospital from April 2015 to November 2015 .The detection results were statistically analyzed .Results The deficiency rates of Cu ,Zn ,Ca ,Mg and Fe in Chengdu area were 0 .4% ,5 .0% ,18 .1% ,9 .5% and 8 .3% respectively ,in which the deficiency rates of Ca in children aged <1 ,1 ,2 ,3 ,≥ 4 years old were 47 .4% ,27 .5% ,12 .6% ,9 .9% and 11 .9% respectively ,indicating that the Ca deficiency rate in children < 1 years old was higher .The Ca deficiency rate had statistical differ‐ence among different age groups(P<0 .05) ,and there were no statistically significant differences in the deficiency rates of several trace ele‐ments among different age groups of male and female children .Conclusion The abnormality rate of trace elements among children aged 0-6 years old in Chengdu area is higher ,in which the Ca deficiency rate is highest ,meanwhile the Mg and Fe were lack too .The trace element content in children is closely related with the feeding habit .The 0-6 years old children in this area should pay attention to the supplement of trace elements ,especially supplement of Ca ,meanw hile breastfeeding is advocated .

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