1.The negative effects of Trim34α on activation of luciferase reporter gene containing NF-κB promoter
Dakang SUN ; Zhenguo SU ; Xinye AN ; Rongjiao ZHOU ; Xiangqin SONG ; Yanting ZHAO
Chinese Journal of Microbiology and Immunology 2011;31(6):492-497
Objective To investigate the effects of Trim34α on the activation of luciferase reporter gene containing NF-κB promoter induced by adaptor proteins TAB2. Methods The total RNA was isolated from HeLa cells. After amplification with RT-PCR, the target sequences were cloned into 5'-Flag-pcDNA3.1 (+) vector. The recombinant vector was confirmed by restriction enzyme digestion, colony PCR and sequencing. It was transfected into HEK293T cells to detected Trim34α expression by Western blot. Simultaneously, the effects of Trim34α on the NF-κB activation induced by TAB2 were determined by dual-luciferase reporter assay. Results Restriction enzyme digestion, colony PCR and sequencing confirmed the vector was constructed successfully, furthermore it expressed Trim34α protein in HEK293T cells. Moreover, trim34α could form high-molecular-weight oligomeric protein, and here we called it trimsome. Interestingly, dual-luciferase assay showed that Trim34α could effectively block TAB2-induced NF-κB activation. Conclusion Trim34α was involved in negative regulation of TAB2-induced NF-κB activation and could form high-molecular-weight oligomer.
2.Evaluating the effect of early clinical exposure in improving medical students' cognitive aspects of the doctor-patient relationship
Huaxiang ZHAO ; Qiang LYU ; Rui ZHANG ; Beilei LIANG ; Yaqi FAN ; Wenzhi DU ; Liyuan Wang ; Xiangqin KONG
Chinese Journal of Medical Education Research 2014;(9):952-957
Objective This paper explored the effect of the early clinical exposure in im-proving medical students' cognitive aspects of the doctor-patient relationship. Methods (1)From 280 undergraduate students of Grade 2010 who participated in early clinical exposure of Xi'an Jiaotong University College of Medicine and 280 undergraduate students of Grade 2011 who did not participate in the early clinical exposure, we selected 140 students respectively to conduct a simple random sam-pling survey. The results were analyzed by SPSS after using Microsoft Excellsoftware for entry. Statis-tical methods selected χ2-test/Fisher exact test(P<0.05). (2)We had an interview to students,teachers leading the project, instructor,experts engaging in the study of the doctor-patient relationship by using focus group discussion and in-depth interview, and then using thematic analysis to analyze the data. (3)The main aspects of questionnaires and interviews were: details, activity participation/interest, general cognitive on the doctor-patient relationship, passion for profession, choice of career, the effect in improving medical students' cognitive aspects of the doctor-patient relationship and the suggestions and comments. Results The effective questionnaires collected from Grade 2010 were 134, and124 from Grade 2011.The participation rate of the undergraduates of Grade 2010(participating group) was 94.0%(126/134), while the interest rate of undergraduates of Grade 2011 (control group) was only 76.6%(95/124). Participating group had a better cognitive understanding of doctor-patient relationship than control group(P=0.0192). The activities had significant effects on choice of career(P=0.0002), and no effect on passion for profession(P=0.7372). There was statistically significant difference on their views of employment(P=0.0002). The key words for the interview were: not enough preparation before activity, not reasonable timing, teachers leading the project, stimulation of professional pride. Conclusions Early clinical exposure activities can be effective in improving medical students' awareness of the current doctor-patient relationship. Still we have some shortages in the activity, some more exploration and amelioration should be made in late stage.
3.Study on mechanism of ginsenoside Rg1-induced human neural stem cells differentiation by genechip.
Xiangqin ZHAO ; Yingbo LI ; Yinghong JIANG ; Di CHEN ; Rong JIANG ; Shali WANG
China Journal of Chinese Materia Medica 2012;37(4):515-519
OBJECTIVEThe molecular targets of ginsenoside Rg1-induced neural stem cells (NSCs) differentiation were screened by genechip.
METHOD7th day following ginsenoside Rg1 induced human neural stem cells to neurons the gene expression was observed by genechip. The purpose gene and signal transduction pathways were selected by the data calculations, and then confirmed by western blot and immunohistochemical method.
RESULT7th day following Rg1-induced NSCs differentiation, there were about 675 different genes, 255 genes of which were up-regulated and 420 genes down-regulated obviously. Meanwhile the ERK1/2 (extracellular signal-regulated protein kinase) in MAPK (mitogen-activated protein kinase) pathway was related with the NSCs differentiation. The Western blot and immunohistochemistry detection confirmed that ERK 1/2 protein and its phosphorylation were significantly increased, which can be blocked by PD98059 (ERK1/2 inhibitor). In addition, differentiation rate of NSCs was also decreased obviously in ginsenoside Rg1-induced differentiated NSCs when ERK blocker PD98059 was used.
CONCLUSIONERK1/2 is an important molecular target in ginsenoside Rg1-induced NSC differentiation. The selected differentially expressed genes by genechip may provide new clues to study of ginsenoside Rg1-induced NSCs differentiation.
Cell Differentiation ; drug effects ; Cell Line ; Down-Regulation ; drug effects ; Flavonoids ; pharmacology ; Ginsenosides ; pharmacology ; Humans ; MAP Kinase Signaling System ; drug effects ; genetics ; Neural Stem Cells ; cytology ; drug effects ; metabolism ; Oligonucleotide Array Sequence Analysis ; Protein Kinase Inhibitors ; pharmacology ; Time Factors
4.The effect of assertive case management on relapse and health economic evaluation in schizophrenics living in communities
Jing ZHAO ; Qinyu LV ; Xiangqin GUO ; Guoqin HU ; Zuowei WANG ; Yaqin JIANG ; Jian CHEN ; Jubao LU ; Yi WU ; Hongtao XIE ; Zhenghui YI
Chinese Journal of Nervous and Mental Diseases 2014;(11):666-672
Objective To evaluate the effect of assertive case management on relapse and health economic evalua?tion in schizophrenics living in communities. Methods Two hundred outpatients were randomly divided into the study group (107 enrolled, 107 completed) which received assertive case management and the control group (93 enrolled, 91 completed) which received normal management treatment for 12 months. Clinical global impression scale (CGI) and the cost of treatment were assessed every three months. Medication adherence and family burden were evaluated before treat?ment and 3, 6 and 12 months after the treatment using Medication Adherence Rating Scale (MARS) and Family Burden Instructing, respectively. Results The study group was less likely to relapse compared with the control group over the 12-month follow-up and the relapse rates were 1.9%and 11.0%in study and control groups, respectively (P<0.01). The repeated-measures analysis of variance indicated that time main effect was significant in severity of illness factor score of CGI (P<0.01). The time main effect and group main effect in factor 1 and factor 3 scores of MARS were significant (all P<0.05) and there was an interaction effect in factor1 score of MARS (P<0.01). In the study group, time effect were signifi?cant in factor 1 score of MARS (P<0.01). The time main effects in indirect cost and total cost were significant and so were interaction effects in direct cost and total cost (P<0.05). In the study group, time effects were significant in direct cost and total cost (P<0.01). Comparison of FBI dimensions before and after the intervention showed that family relation?ship was much more decreased in the study group than in the control group (P<0.01). Conclusions Assertive Case Man?agement can reduce the recurrence of schizophrenia living in communities, improve compliance medication and family re?lationship as well as reduce the cost of treatment.
5.Effect of miR-125a-5p targeting Scarb1 gene on hypoxia/reoxygenation injury of cardiomyocytes and its mechanism.
Yintao ZHAO ; Jingjing SUN ; Yuan LIU ; Lu ZHENG ; Xiangqin ZHANG ; Haibo YANG
Chinese Journal of Medical Genetics 2020;37(9):980-986
OBJECTIVE:
To explore the effect and mechanism of miR-125a-5p targeted regulation of scavenger receptor B1 (Scarb1) gene on anoxia/reoxygenation injury of rat cardiomyocytes.
METHODS:
H9c2 rat cardiomyocytes were randomly divided into blank control group, hypoxia/reoxygenation group, transfection control group and mir-125a-5p transfection group. The expression of miR-125a-5p, cardiomyocyte viability, apoptosis rate, ATP content and the expression of Scarb1, Cyt C, Bax, Bcl-2 and NF-κB signaling pathway related proteins were determined. Target gene of miR-125a-5p was predicted with Targetscan software, and the targeting of miR-125a-5p on Scarb1 was verified by double luciferase reporter gene experiment.
RESULTS:
Compared with the blank control group, the expression of miR-125a-5p, Bax, Cyt C and the apoptotic rate of cardiomyocytes in the hypoxia/reoxygenation group were significantly increased (P<0.05), while the expression of Scarb1, Bcl-2 and the content of ATP were significantly decreased (P<0.05). Compared with the control group, the situation of mir-125a-5p transfection group was just the opposite. Double luciferase reporter gene experiment has confirmed Scarb1 to be the target of miR-125a-5p. Hypoxia/reoxygenation can promote the expression of NF-κB p65, C-myc and Cyclin D1 in cardiomyocytes, while down-regulating the expression of miR-125a-5p can inhibit the expression of such proteins.
CONCLUSION
Hypoxia/reoxygenation can induce the expression of miR-125a-5p in rat cardiomyocytes. Inhibition of miR-125a-5p can protect cardiomyocytes from hypoxia/reoxygenation by up-regulating the expression of Scarb1. The mechanism may be related to the inhibition of activation of NF-κB signaling pathway.
6.Multicenter ultrasound screening for the results of carotid atherosclerotic lesions in a Chinese population with high-risk of stroke:a preliminary analysis
Yang HUA ; Yunlu TAO ; Mei LI ; Qiang YONG ; Wen HE ; Hui ZHAO ; Yan LUO ; Yan ZHANG ; Tao PENG ; Delin YU ; Xudong PAN ; Chunxia WU ; Xiaoyuan NIU ; Fengyun HU ; Xiangqin HE ; Jianjun YUAN ; Wen CHU ; Fengzhen TANG ; Hong AI ; Jinchuan CUI
Chinese Journal of Cerebrovascular Diseases 2014;(12):617-623
Objectives To screen the high-risk population of stroke in China using color Doppler flow imaging (CDFI)and to establish a stroke risk prediction model in Chinese population in order to prevent and treat stroke early. Methods Forty-one base hospitals and 715 286 people in the project areas of the first 6 provinces of China conducted routine physical examinations and investigated the related risk factors for cardiocerebrovascular diseases from July 2011 to April 2012 using a cross-sectional study,among them 61 860 patients underwent carotid CDFI screening,and 49 386 of them were high-risk population (exposed to≥3 risk factors). The bilateral common carotid interma-media thickness (IMT),the number of plaques and the degree of carotid stenosis were screened and documented. And whether carotid IMT thickening or not,with or without carotid plaques,and degree of carotid artery stenotic rate 0-49% and≥50% were performed by multivariate logistic regression analysis with the risk factors for stroke,respectively. Results (1)Logistic regression analysis showed that hypertension,atrial fibrillation,smoking,and lack of physical exercise were the independent risk factors for carotid IMT thickening (hypertension:OR,1. 17;95%CI 1. 12-1. 22;atrial fibrillation:OR,1. 15;95%CI 1. 09-1. 21;smoking:OR,1. 13;95%CI 1. 08-1. 17;and lack of physical exercise:OR,1. 12;95%CI 1.08-1. 16). (2)Hypertension,atrial fibrillation, smoking,and diabetes were the independent risk factors for carotid plaque and carotid artery stenosis rate≥50%(carotid plaque,hypertension:OR,1. 55;95%CI 1. 47-1. 62;atrial fibrillation:OR,1. 13;95%CI 1.06-1. 21;smoking:OR,1. 16;95%CI 1. 11-1. 22;and diabetes:OR,1. 30;95%CI 1. 24-1. 37). Carotid stenosis rate≥50%,hypertension:OR,1. 78;95%CI 1.55-2. 03;atrial fibrillation:OR,1. 59;95%CI 1. 39-1. 81;smoking:OR,1. 33;95%CI 1. 20-1. 48;and diabetes:OR,1. 30;95%CI 1. 17-1. 45. Simple obesity did not increase the incidences of carotid atherosclerotic plaque and carotid artery stenosis ≥50%(OR,0. 78, 0.83;95%CI 0. 75-0. 82 ,0. 75-0. 92,respectively). Conclusions Neck vascular ultrasound can be used as a valuable means for screening high-risk population and detecting risk factors of stroke. It has an important clinical significance for the early diagnosis and treatment of carotid atherosclerosis disease.
7.Risk factors for early acute kidney injury after classic orthotopic liver transplantation
Jiahong CHEN ; Ying XU ; Songzhe HE ; Tao LI ; Yunhao BAI ; Xiangqin SONG ; Tiantian ZHAO ; Wei LI ; Hongtao JIANG ; Yi WANG
Chinese Journal of Organ Transplantation 2023;44(5):269-274
Objective:To explore the risk factors affecting the incidence of acute kidney injury(AKI)after liver transplantation(LT).Methods:From November 2019 to November 2022, clinical data were retrospectively reviewed for 105 recipients of classic orthotopic LT.There are 89 males and 16 females with an age range of(50.52±10.35)years.They are assigned into two groups of AKI(66 cases)and non-AKI(39 cases)according to the AKI diagnostic and staging criteria of Global Kidney Disease Prognosis Organization in 2012.General profiles and clinical data(e.g.previous medical history, MELD score, total bilirubin, albumin, serum creatinine level, coagulation function, anhepatic phase and time to surgery)of two groups of recipients are compared.The factors with statistically significant differences are included into multivariate Logistic regression analysis for obtaining independent risk factors for early AKI post-LT.Results:Among them, 66 patients developed AKI within 7 days post-operation with an incidence rate of 62.86%(66/105).The clinical stages of AKI are Ⅰ(46 cases, 69.70%), Ⅱ(10 cases, 15.15%)and Ⅲ(10 cases, 15.15%).Statistically significant inter-group differences exists in age, abdominal surgery history, preoperative serum level of creatinine, operative duration, anhepatic phase and intraoperative plasma transfusion(all P<0.05).Multivariate Logistic regression analysis indicated that abdominal surgery history( OR=5.803, 95% CI: 1.008~33.401, P=0.049), anhepatic phase( OR=1.054, 95% CI: 1.008~1.101, P=0.020)and preoperative serum level of creatinine( OR=0.968, 95% CI: 0.943~0.994, P=0.016)are independent risk factors for early AKI after classical orthotopic LT recipients. Conclusions:Abdominal surgery history, anhepatic phase, and preoperative serum level of creatinine are independent risk factors for early AKI in classic orthotopic LT recipients.
8.Prokaryotic expression of fowl adenovirus serotype 11 Fiber protein and prepara-tion of its polyclonal antibody
Shuaifeng LIU ; Yifan MA ; Xiangqin WANG ; Xiaoran GUO ; Xiaona LIU ; Baishi LEI ; Wanzhe YUAN ; Fuxing CHEN ; Kuan ZHAO
Chinese Journal of Veterinary Science 2024;44(7):1387-1393
In order to obtain polyclonal antibodies against the fibrillar(Fiber)protein of fowl ade-novirus serotype 11(FAdV-11)and investigate its cross-reactivity to different serotypes of FAdV Fiber,the gene encoding the FAdV-11 Fiber protein was cloned into a prokaryotic expression vec-tor pET-32a by homologous recombination technology,then the plasmid was transformed into BL21(DE3)receptor cells,and the purified recombinant protein was used as an immunogen to im-munize rabbits to prepare polyclonal antibody after induced expression,and the cross-reactivity of the polyclonal antibody against different serotypes of FAdV Fiber proteins was identified by West-ern blot and indirect immunofluorescence(IFA).The results showed that the His-FAdV-11-Fiber recombinant protein was mainly expressed as inclusion bodies and was well expressed.Western blot and IFA showed that the prepared polyclonal antibody reacted with the Fiber proteins of FAdV-8a,FAdV-8b,and FAdV-11,but did not with the 2 Fiber of FAdV-4(Fiber 1 and Fiber 2)proteins.In conclusion,in this study,we successfully prepared rabbit polyclonal antibodies against FAdV-11 Fiber and showed that it specifically recognized the Fiber proteins of FAdV-8a,FAdV-8b and FAdV-11,which lays the foundation for further establishment of serological differential diag-nosis of FAdV-11.