1.Effect of epidermal growth factor on the expression of matrix metalloproteinase-9 and the signalling pathways involved in the trophoblast cell line JEG-3
Huaibin REN ; Ziyan JIANG ; Lizhou SUN ; Mingsong FAN ; Yanfen ZOU
Chinese Journal of Obstetrics and Gynecology 2011;46(7):521-526
Objective To investigate the effect of epidermal growth factor (EGF) on the expression of Matrix metalloproteinase-9 (MMP-9) and the signalling pathways involved in the trophoblast cell line JEG-3. Methods The JEG-3 trophoblast cell line was used in this study. (1) JEG-3 cells were cultured with various concentrations of EGF (0, 1, 10,20 ng/ml) for 24 hours and the expression of MMP-9 was tested by western blotting and reverse transcription PCR (RT-PCR). (2) Western blotting and RT-PCR were also used to investigate the expression of MMP-9 expression after incubation for 0,4,12 and 24 hours with EGF treatment (10 ng/ml) in JEG-3 cells. (3) According to the different added ingredients, JEG-3 cells were divided into some groups: control group (without EGF), EGF group (exposure to l0ng/ml EGF),EGF + inhibitors group (exposure to 10 ng/ml EGF +20 ng/ml SB203580 or exposure to 10 ng/ml EGF + 10ng/ml U0126) inhibitors group (exposure to 20 ng/ml SB203580 or exposure to 10 ng/ml U0126). Western blotting were used to investigate the expression levels of MMP-9, nuclear factor kappa B (NF-kB) ,p38MAPK, phospho-p38MAPK (p-p38MAPK) , extracellular -signal regulated kinase (ERK) and phospho-ERK (p-ERK) protein in JEG-3 cells after incubation for 24 hours. Results (1) The profiles of MMP-9mRNA were increased by various concentrations of EGF (0, 1 , 10, 20 ng/ml) in JEG-3 cells after 24hculture. The expression of MMP-9 mRNA in JEG-3 cells exposure at 1 ng/ml of EGF (0. 567 ±0. 056) , 10ng/ml of EGF (1. 392 ±0. 133) , 20 ng/ml of EGF (1. 971 ±0. 067) were significantly higher respectively (P <0. 05) , compared with 0 ng/ml of EGF treatment (0. 166 ±0. 015). Similarly, MMP-9 mRNAs were also increased with the increasing incubation time. Compared to EGF (10 ng/ml) stimulation for 0 h (0.253 ±0.044), the MMP-9 mRNA profiles were 0. 470 ±0. 026, 1.061 ±0. 115, 1. 453 ±0. 180 for 4,12 and 24 hours, respectively (P < 0. 05). (2) In accordance to the mRNA profiles, the expression of MMP-9 protein was also increased by different concentrations of EGF (0,1, 10, 20 ng/ml) in JEG-3 cells after 24 h-culture. The abundance of MMP-9 protein in the three groups was 0. 043 ±0. 012, 0. 085 ±0. 008, 0. 142 ±0. 015, with a significantly higher expression, compared with 0 ng/ml of EGF treatment (0. 004 ±0.001, P < 0.05) respectively. Similarly, MMP-9 proteins were also increased with the increasing incubation time. Compared to EGF(10 ng/ml) stimulation for 0 h (0. 030 ±0. 009) , the profiles of MMP-9 protein were 0. 137 ± 0. 010, 0. 240 ± 0. 010, 1.240 ±0.061 for 4, 12 and 24 hours, respectively (P < 0.05). (3) Both p38MAPK and ERK signalling pathways were activated by EGF in JEG-3 cells. The expression of p-p38MAPK was significantly higher (without or with 10 ng/ml EGF, 234. 1 ± 4. 1 vs.260. 9 ± 2. 5 , P < 0. 05) , however, the p38 MAPK inhibitor SB203580 markedly suppressed the increase in p-p38MAPK content induced by EGF(227. 9 ±2. 4 vs. 260. 9 ±2. 5, P<0. 05). Similarly, the expression of p-ERK was significantly higher with EGF treatment (812. 2 ±3. 5) vs. without EGF group (453.4±5.8) (P <0. 05) , while the ERK inhibitor U0126 significantly inhibited the increased p-ERK content in response to EGF treatment (71. 0 ± 1. 2 vs. 812. 2 ± 3. 5, P < 0. 05) . (4) The p38MAPK inhibitor SB203580 significantly reduced the expression of EGF-induced MMP-9 (0. 645 ± 0. 270 vs. 1. 476 ± 0. 452, P < 0. 05)and NF-kB (0.530 ± 0.026 vs. 0.959 ± 0. 017, P < 0. 05) . (5) The ERK inhibitor U0126 also significantly reduced the expression of EGF-induced MMP-9 (0. 623 ±0. 030 vs. 2. 112 ±0. 056, P <0. 05)and NF-kB (0. 325 ± 0. 082 vs. 0. 939 ± 0. 153, P < 0. 05). Conclusion EGF induced the expression of MMP-9 in a time and dose-dependant manner in JEG-3 cells. EGF enhanced MMP-9 expression through the activation of p38MAPK and ERK signalling pathways in JEG-3 cells.
2.Effect of transforming growth factor β1 on the expression of matrix metalloproteinase 9, tissue inhibitor of metalloproteinase 1 and nuclear factor kappa B signalling pathway in the human amniotic cells WISH
Mingsong FAN ; Ziyan JIANG ; Yanfen ZOU ; Lin QU ; Xue ZHOU ; Lizhou SUN
Chinese Journal of Obstetrics and Gynecology 2013;(1):29-33
Objective To investigate the effect of transforming growth factor β1 (TGF-β1) on the expression of matrix metalloproteinase 9 (MMP-9),tissue inhibitor of metalloproteinase 1 (TIMP-1),nuclear factor kappa B(NF-κB) and the possible signalling pathways in human amniotic cells WISH.Methods The WISH cell line was cultured.WISH cells were added with TGF-β1 of different concentrations (0,2,10 and 20 ng/ml,respectively) for 24 hours.Then,reverse transcription (RT) PCR and western blotting were used to analyze the protein and mRNA expression of TIMP-1 and MMP-9; and the expression of NF-κB was analyzed by western blot.Results (1) The profile of TIMP-1 mRNA (0.413 ±0.036,0.623 ±0.058,1.392 ±0.124,1.387 ±0.102) in WISH cells elevated when the concentration of TGF-β1 increased (0,2,10,20 ng/ml).In accordance with TIMP-1 mRNA,the expression of TIMP-1 also elevated with the increase of TGF-β1 (0.357 ± 0.031,0.596 ± 0.048,1.243 ± 0.097 and 1.359 ± 0.121,respectively).And when 2,10 or 20 ng/ml of TGF-β1 was added,the TIMP-1 mRNA and protein were significantly higher than the TIMP-1 mRNA and protein when no TGF-β1 was added(P < 0.05).(2)In contrast with TIMP-1,MMP-9 mRNA (1.325 ±0.056,0.987 ±0.081,0.610 ±0.034,0.347 ±0.023) in WISH cells decreased when the concentration of TGF-β1 increased (0,2,10,20 ng/ml).The MMP-9 protein (1.119 ±0.064,1.008 ±0.052,0.578 ±0.041,0.401 ±0.015) also decreased with the increase of TGF-β1.And when 2,10 or 20 ng/ml of TGF-β1 was added,the MMP-9 mRNA and protein were significantly lower than the MMP-9 mRNA and protein when no TGF-β1 was added (P < 0.05).(3) The NF-κB protein (1.423 ±0.065,1.116 ± 0.045,0.796 ± 0.041,0.359 ± 0.021) was significandy reduced with the increase of TGF-β1 (0,2,10,20 ng/ml; P < 0.05).Conclusions The mRNA and protein expression of TIMP-1 decreased when TGF-β1 was low in WISH cells,whereas those of MMP-9 elevated when TGF-β1 was low.The unbalance of TIMP-1 and MMP-9 was related to the pathology of the premature rupture of membrane.And the NF-κB singalling pathway might be an important mechanism in the regulation of TIMP-1 and MMP-9 system.
3.Clinical research on COX-2 ,NF-κB and VEGF expression in triple negative breast cancer
Lingdi MA ; Yanfen DONG ; Qian LIU ; Jing FAN ; Lijia JIANG ; Yahong CHEN
International Journal of Laboratory Medicine 2016;37(4):436-437,440
Objective To study the cyclooxygenase (COX-2) ,nuclear factor-κB (NF-κB) and vascular endothelial growth factor (VEGF) expression and clinical significance in triple negative breast cancer .Methods From January 2010 to December 2014 ,breast cancer treatment in our hospital 100 patients for the study ,50 patients with triple negative breast cancer ,50 cases of non-triple neg-ative breast cancer was detected by immunohistochemistry ,100 cases of breast cancer were detected by immunohistochemistry in or-ganizations COX-2 ,NF-κB and VEGF expression of lymphatic vessel invasion and lymph vessel density and D2-40 mark detection , statistical analysis of relevant clinical and pathological information .Results COX-2 in triple negative and non-triple negative breast cancer were 76% ,70% ,the difference was not statistically significant (P>0 .05) ,VEGF triple negative breast cancer and non-triple negative breast lesions in cancerous lesions positive expression rates of 60% and 36% ,respectively ,which had significant difference (P<0 .05) .NF-κB in triple negative breast cancer lesions and non-triple negative breast lesions positive expression rate was 66%and 32% ,respectively ,which had significant difference (P< 0 .05) .Triple negative breast cancer NF-κB and VEGF ,COX-2 and VEGF expression was significantly positively related to breast cancer .Conclusion Radiation and chemotherapy is a major means of triple negative breast cancer postoperative treatment ,while inhibiting the NF-κB ,the expression of VEGF and COX-2 is expected to become the new target for treatment of triple negative breast cancer ,is worth exploring .
4.Effects of different frequencies of pricking-bloodletting at auricular points plus auricular point sticking therapy on the serum levels of related factors in patients with acne vulgaris
Man ZHANG ; Mingjian ZHANG ; Yanfen SHE ; Yayu GAO ; Hao CHEN ; Jin LIU ; Xisheng FAN ; Jun LIU ; Juncha ZHANG
Journal of Acupuncture and Tuina Science 2023;21(1):74-81
Objective: To observe the effects of different frequencies of pricking-bloodletting at auricular points plus auricular point sticking therapy on the levels of the serum immunoglobulin G (IgG), immunoglobulin M (IgM), testosterone (T), and insulin-like growth factor-1 (IGF-1) in patients with acne vulgaris (AV). Methods: Ninety patients with AV were randomly assigned to treatment group 1, treatment group 2, and treatment group 3 according to the random number remainder grouping method, with 30 cases in each group. All three groups received pricking-bloodletting at auricular points plus auricular point sticking therapy. Treatment group 1 was treated once a week, treatment group 2 was treated twice a week, and treatment group 3 was treated 3 times a week. Four-week treatment was taken as 1 treatment course, and 3 treatment courses were observed. On the day before the start of the study and on the day next to the end of each course, the global acne grading system (GAGS) score was recorded, and 3 mL of blood from the median cubital vein was collected to test the serum levels of IgG, IgM, T, and IGF-1. After 3 courses of treatment, the efficacy index was calculated according to the GAGS score, and the serum indicators in patients with effective treatment (efficacy index ≥20%) and complete sampling were analyzed. Results: A total of 60 patients were included in the final blood indicator analysis. After 3 courses of treatment, the intra-group comparisons showed that the serum IgG level increased significantly in patients in treatment group 1 (P<0.01); the serum T level decreased significantly in the female patients in treatment group 2 (P<0.05); the IGF-1 level significantly decreased in the patients in all three groups (P<0.05 or P<0.01). There were no significant differences in the serum levels of IgG, IgM, T, and IGF-1 among the three groups at each time point (P>0.05). Conclusion: Pricking-bloodletting at auricular points plus auricular point sticking therapy can affect the levels of serum IgG, T, and IGF-1 in AV patients. The level of the serum IGF-1 can be reduced by treatment once a week, twice a week, or 3 times a week. Treatment once a week can increase the patients' serum IgG level, and treatment twice a week can significantly decrease the serum T level in female patients. Reducing the serum IGF-1 level may be one of the action mechanisms of pricking-bloodletting at auricular points plus auricular point sticking therapy in treating AV.
5.Seeking specific response points from the three Yin meridians of foot using laser speckle contrast imaging in patients with primary dysmenorrhea
Xisheng FAN ; Panpan WEI ; Xuliang SHI ; Xiaodan SONG ; Mingjian ZHANG ; Juncha ZHANG ; Jun LIU ; Lijia PAN ; Xiaoyi DU ; Yanfen SHE ; Jue HONG
Journal of Acupuncture and Tuina Science 2023;21(5):405-412
Objective:To seek specific response points on the body surface of patients with primary dysmenorrhea(PD)by observing blood perfusion unit(PU)at different points of the three Yin meridians of foot using laser speckle contrast imaging(LSCI). Methods:Eighty PD patients were recruited as a PD group,and 80 healthy female undergraduates were taken as a normal group.During one menstrual cycle(before menstruation,during menstruation,and 3 d after menstruation),each participant was examined using the LSCI system to determine PU at bilateral Taixi(KI3),Taibai(SP3),Taichong(LR3),Shuiquan(KI5),Diji(SP8),Zhongdu(LR6),Sanyinjiao(SP6),and Xuehai(SP10)and non-acupuncture points.The researchers in charge of point location,operation,and statistical analysis were not aware of grouping.PU at the detection spots was taken as the outcome measure. Results:Compared with the normal group,the PD group showed increases in PU at right Taixi(KI3)before menstruation(P<0.05)and at bilateral Zhongdu(LR6)and right Diji(SP8)during menstruation(P<0.05).At the other time points,significance was not found between the two groups in comparing PU at the detected spots. Conclusion:Compared with healthy participants,PD patients present specific changes in PU at Taixi(KI3),Diji(SP8),and Zhongdu(LR6)at specific time points during the menstrual cycle,which provides a reference for acupuncture-moxibustion treatment of PD in clinical settings.
6.Effect of arotinolol on right ventricular function in patients with dilated cardiomyopathy
Hong YANG ; Li XU ; Yongkang TAO ; Zhimin XU ; Xiuqing DU ; Naqing LU ; Jinglin ZHAO ; Xianqi YUAN ; Yanfen ZHAO ; Rongfang SHI ; Chaomei FAN
Journal of Geriatric Cardiology 2007;4(3):170-173
Objective Dilated cardiomyopathy (DCM) is generally considered to be accompanied by both left and right ventricular dysfunction,but most studies only analyze the left ventricular function. In this study, we evaluated the effect of arotinolol on right ventricular function in patients with DCM. Methods Right ventricular ejection fraction (RVEF) and right ventricular diameter (RVD) were measured by two-dimensional echocardiography (2-DE) in 33 DCM patients; RVEF measured by first-pass radionuclide angiography (FPRA) was compared with that by 2-DE. Results The treatment with arotinolol for one year resulted in a reduction in the right ventricular diameter (baseline, 23.0 ± 8.3 mm vs after one-year treatment, 20.7 ± 5.4 mm; P=0.004 ) and an associated increase in ejection fraction (baseline, 36.9 ± 10.3% vs after one-year treatment, 45.8 ± 9.6%; P < 0.001 ); there is a high correlation between the 2-DE method and radionuclide ventriculographic method. The correlation coefficient is 0.933 (P<0.001). Conclusion Arotinolol therapy could not only improve left ventricular function, but also improve right ventricular function in DCM patients.
7.Studies on the correlation of 489 cases of bronchial asthma complicated with allergic rhinitis in Datong.
Yanfen BIAN ; Yaoguo ZHU ; Shangde GUO ; Jianbin WANG ; Zhilin XUE ; Xuemin FAN ; Jie ZHANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2009;23(2):67-69
OBJECTIVE:
To discuss the disease incidence of bronchial asthma (BA) complicated with allergic rhinitis (AR) and the correlation of their age, classification, concomitant symptom.
METHOD:
Four hundred-nine cases of AR were identified by means of random sampling, physical examination, laboratory test and questionnaire. According to the diagnostic criteria of BA, they were divided into two groups: group I ( control group) including 298 cases suffering from only AR, group II ( experimental group) 191 cases of BA concomittent with AR.
RESULT:
1) This study showed that BA concomittent with AR account for 39% of all cases; 2) Incidence of aspirin triad syndrome (asthma, rhinopolyps and aspirin intolerance) was significantly different in two groups (P<0.01); 3) As for age group distribution, there was significant difference between groups in age segment 10-19 and 40-49 (P<0.05); 4) Disease classification: there were significant differences in the incidence of moderate to severe intermittent AR, mild continuous AR, and moderate to severe continuous AR between two groups (P<0.05); 5) Other major concomitant diseases and symptoms distribution; the concomittant occurrence of allergic pharyngitis, sinusitis, conjunctivitis, secretory tympanitis, hypertrophic rhinitis, dermatosis, discomfort of gastrointestinal tract, and headache of unknown origin were significantly different between two groups (P<0.05). The difference above showed that the occurrence in experimental group was higher than that in control group.
CONCLUSION
the incidence of BA complicated with AR were relatively high in Datong, with a peak incidence at age 10 to 19 and 40 to 49. The common types of disease were moderate to severe intermittent AR, mild continuous AR, and moderate to severe continuous AR. The number of patients with BA complicated with AR were growing.
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8.Analysis of global research hotspots in the field of pediatrics based on Essential Science Indicators and its revelation
Yanfen FAN ; Jianhua LI ; Weihua SHAN ; Yumei SHEN ; Zhenzhen LIAN ; He ZANG ; Jiaqin WANG ; Hongling FANG
Chinese Journal of Applied Clinical Pediatrics 2022;37(22):1756-1760
Objective:To analyze the global research hotspots in the field of pediatrics based on the Essential Science Indicators (ESI) database and explore the inspiration to domestic editors and pediatrics researchers.Methods:The journal distribution, country (region) distribution, cooperation, organization distribution, funding, publication language, hot topic words and other data of highly cited papers in the field of pediatrics in ESI database were collected and analyzed.Results:A total of 682 highly cited pediatrics papers were collected from 77 pediatrics journals included in Science Citation Index(SCI). Most of the highly cited pediatrics papers (182) were found to be published in Pediatrics.All 682 paper were published in English and frequently, characterized by multiple authors, institutions and fund support.Of 682 highly cited pediatrics papers, 435 papers were published in the United States(the first), 123 papers in England(the second) and 86 paper in Canada(the third). Novel coronavirus pneumonia, coronavirus, SARS coronavirus, autism and multiple system inflammatory syndrome are the main frontiers of global pediatric research at present.Specifically, focal pediatric system diseases mainly include respiratory system diseases, digestive system diseases, cardiovascular diseases, etc. Conclusions:ESI-based analysis of global research hotspots in the field of pediatrics provides reference materials for domestic and foreign pediatrics researchers to understand the global academic frontiers and development trends in the field of pediatrics and select topics for future scientific research.More importantly, this analysis can help domestic editors of pediatrics journals to plan topics and organize hot papers, so as to improve the academic quality and international influence of the journals.
9.Preliminary study on the value of pericoronary adipose tissue histogram parameters for the differentiation of acute coronary syndrome and stable coronary artery disease
Qing TAO ; Weijie ZOU ; Yanfen FAN ; Hailin SHEN ; Hongdi DU ; Chunhong QIAN ; Feng ZHU ; Su HU ; Guangyu HAO ; Shaofeng DUAN ; Chunhong HU
Chinese Journal of Radiology 2020;54(3):192-197
Objective:To investigate the value of pericoronary adipose tissue histogram parameters based on coronary CT angiography (CTA) images for the differentiation of acute coronary syndrome and stable coronary artery disease.Methods:The clinical data and CTA images of 93 patients with coronary CTA examination in Suzhou Kowloon Hospital from 2013 to 2018 were analyzed retrospectively. There were 39 patients with acute coronary syndrome (acute coronary syndrome group) and 54 patients with stable coronary artery disease (stable coronary artery disease group). A region of interest (ROI) was drawn around the stenosis of the coronary arteries, with CT attenuation ranging from-190 to -30 HU to exclude non-adipose tissue. The CT attenuation of ROI excluding non-adipose were measured and histogram analysis was performed. The obtained parameters included the mean value, median value and the 5th, 10th, 45th, 55th, 70th and 95th percentiles. The differences in histogram parameters between the two groups were compared, and then the value of each parameter in differentiating acute coronary syndrome and stable coronary artery disease was evaluated based on receiver operating characteristic (ROC) analysis. The stepwise regression of multivariate logistic regression analysis was used to identify the useful features and establish the final prediction model. The ROC curve of the final model was calculated and its value was analyzed.Results:The mean, median and the 5th, 10th, 45th, 55th,70th and 95th percentile differences between the acute coronary syndrome group and the stable coronary artery disease group were statistically significant (all P<0.05). The ROC curve for the median and the 95th percentile had the same area under curve (AUC) of 0.73. The sensitivity, specificity and AUC of the diagnostic model established by multiple logistic regression were 82.1%, 89.1% and 0.90 respectively. Conclusion:CT attenuation histogram of pericoronary adipose tissue is of high value in differentiating acute coronary syndrome from stable coronary artery disease.
10.The value of gadolinium-ethoxybenzyl-diethylenetriamine pentaacetic acid enhanced MRI nomogram model for predicting Ki-67 expression of hepatocellular carcinoma
Mengjie HU ; Yixing YU ; Yanfen FAN ; Ximing WANG ; Su HU ; Chunhong HU
Chinese Journal of Radiology 2020;54(12):1185-1190
Objective:To explore the value of gadolinium-ethoxybenzyl- diethylenetriamine pentaacetic acid (Gd-EOB-DTPA) enhanced MRI nomogram model for preoperative prediction of Ki-67 expression in hepatocellular carcinoma (HCC).Methods:Data of 85 patients of HCC confirmed by postoperative pathology, who underwent preoperative Gd-EOB-DTPA enhanced MRI between September 2016 and August 2019 in the First Affiliated Hospital of Soochow University were retrospectively evaluated. According to postoperative immunohistochemistry Ki-67 index, the 85 patients were divided into Ki-67 low expression group(Ki-67 index ≤10%, n=20) and Ki-67 high expression group (Ki-67 index >10%, n=65). Clinical data (hepatitis, cirrhosis, etc.), qualitative imaging parameters (tumor margin, capsule, etc.) were compared by χ 2 test and quantitative parameters [lesion-to-normal parenchyma ratio-arterial phase (LNR-AP), lesion-to-normal parenchyma ratio-portal phase (LNR-PP), lesion-to-normal parenchyma ratio-equilibrium phase (LNR-EP) and lesion-to-normal parenchyma ratio-hepatobiliary phase (LNR-HBP)] were compared by independent sample t test. The above statistically significant parameters were included in multivariate logistic regression to identify the independent predictors of Ki-67 high expression and then the nomogram model for predicting Ki-67 expression of HCC was established. Results:alpha-fetoprotein (AFP) tumor margin, arterial rim enhancement between the Ki-67 low expression group and the Ki-67 high expression group had significant differences (χ 2 were 8.196, 10.538 and 4.717, respectively, P<0.05). LNR-AP, LNR-PP, LNR-EP and LNR-HBP between the two groups had significant differences ( t were 2.929, 2.773, 2.890 and 3.437, respectively, P<0.05).The result of multivariate logistic regression revealed that AFP≥20 μg/L, non-smooth tumor margin and low LNR-HBP were the independent predictors of Ki-67 high expression (odds ratio were 4.090, 3.509 and 0.042, respectively, P<0.05).The Gd-EOB-DTPA enhanced MRI nomogram model for predicting Ki-67 expression of HCC was established successfully. The Area under the receiver operating characteristic curve of the nomogram was 0.837 and the corrected predictive curve fitted the ideal curve, which suggested the model had a good predictive efficiency. Conclusion:Gd-EOB-DTPA enhanced MRI nomogram model has great value in preoperative prediction of Ki-67 expression of HCC, which provided a personalized prediction method for Ki-67 expression in patient with HCC.