2.Regulatory Effects of miRNA-31 on LATS2 and Cardiomyocyte Hypertrophy in Rat's Cardiomyocytein vitro
Junyi ZENG ; Wan ZHANG ; Lu DING ; Yunfeng WEI ; Zeqi ZHENG ; Tong WEN ; Yongnan FU
Chinese Circulation Journal 2017;32(2):177-182
Objective: To observe the regulatory effects of miRNA-31 (miR-31) on LATS2 and cardiomyocyte hypertrophy via down-regulating miR-31 expression in rat's cardiomyocytesin vitro. Methods: Rat's cardiomyocytes were isolated and cultured for 10 daysin vitro, according to different intervention methods, the cells were divided into 4 groups:①Blank control group,②AngII intervention group,③Lentivirus with miR-31 inhibitor infection group,④Negative lentivirus infection group. On day-8, gene expressions of MiR-31, LATS2, cardiac hypertrophy ANP and β-MHC were examined by qRT-PCR; on day-10, cell morphology was observed by fluorescence staining. LATS2 protein expression was examined by Western blot analysis. Dual luciferase reporter plasmids were transfected into 293T cells, then luciferase activity was detected to identify the targeting effect of miR-31 on LATS2. Results: Compared with Blank control group, AngII intervention group showed increased gene expressions of miR31, cardiac hypertrophy ANP and β-MHC,P<0.05, enlarged cardiomyocyte surface,P<0.05; while decreased gene and proteinexpressions of LATS2,P<0.05. Compared with AngII intervention group, Lentivirus with miR-31 inhibitor infection group had down-regulated expressions of miR31, cardiac hypertrophy ANP and β-MHC,P<0.05, reduced cardiomyocyte surface, P<0.05; while slightly increased LATS2 gene expression and obviously increased protein expression,P<0.05. Dual luciferase reporter assay presented that relative luciferase activity of TRAF6-3' UTR+miR-146b was significantly decreased than TRAF6-3' UTR+miR-NC,P<0.01 and relative luciferase activity of LATS2-3' UTR+ miR-31 was signiifcantly reduced than LATS2-3' UTR-NC+miR-31,P<0.01. Conclusion: Cardiomyocytes hypertrophy could be reversed at certain degree by down-regulating miR-31; the targeting effect of miR-31 on LATS2 was involved in cardiomyocyte hypertrophyregulation.
3.Genetic recombinant prokaryotic expression of augmenter of liver regeneration and preparation of polyclonal antibodies in rats
Zhengfang LIU ; Jianming WANG ; Lan WANG ; Xiaoyun ZENG ; Ling XIONG ; Zhixiu LUO ; Junyi WU
Chinese Journal of Tissue Engineering Research 2010;14(11):1943-1947
BACKGROUND:An abroad study repoRed the distribution and expression of augmenter of liver regeneration(ALR)in the central nervous system.There are few literatures on how to prepare and evaluate ALR protein polyclonal antibody in recombinant rats,and how to construct prokaryotic expression vector.There are no repots concerning ALR in the central nervous system in China.OBJECTIVE:TO express ALR fusion protein in E coli BL21 and prepare and identify polyclonal antibody.METHODS:RNA was extracted from the hippocampus of Sprague Dawley rats.The prokaryotic expression plasmid pET28a-ALR was constructed and the positive recombinant plasmid was transformed into BL21.Protein ALR was expressed by inducing transformed BL21 with Isopropyl-β-D-thiogalactopyranoside(IPTG)and purified by Ni~(2+)affinity chromatography column after immune the rabbit for 4 times.the serum of rabbits was extracted from hear as polyclonal antibody.The titer and specificity of the rabbit's antiserum was respectively measured by ELISA and Western blotting The following parameters were measured:construction of prokaryotic expression plasmid pET26a-ALR;pET28a-ALR recombinant enzyme digestion evaluation;results of ELISA and Western-blotting.RESULTS AND CONCLUSION:Expecting bands were obtained by double enzyme digestion electrophoresis,respectively 5.3 kb and 0.4 kb.Nucleotide sequence analysis verified that prokaryotic expression vector pET28a-ALR was successfully constructed.The 19 ku fusion protein was successfuIly expressed.The titer of the antiserum measured by ELlSA could achieve 1:2 000 This indicated that antibody and purified recombinant ALR had a good reaction.and high titer.could meet the experimental require.Western blotting analysis proved that the antibody could identify the prokaryotic expression product of ALR.Prokaryotic expression system expressed ALR fusion protein,prepared and purified polyclonal antibody of ALR protein,and could meet the experimental require of ALR immunoblotting.
4.Elastic registration method to compute deformation functions for mitral valve.
Jinyu YANG ; Wan ZHANG ; Ran YIN ; Yuxiao DENG ; Yunfeng WEI ; Junyi ZENG ; Tong WEN ; Lu DING ; Xiaojian LIU ; Yipeng LI
Journal of Biomedical Engineering 2014;31(5):1135-1138
Mitral valve disease is one of the most popular heart valve diseases. Precise positioning and displaying of the valve characteristics is necessary for the minimally invasive mitral valve repairing procedures. This paper presents a multi-resolution elastic registration method to compute the deformation functions constructed from cubic B-splines in three dimensional ultrasound images, in which the objective functional to be optimized was generated by maximum likelihood method based on the probabilistic distribution of the ultrasound speckle noise. The algorithm was then applied to register the mitral valve voxels. Numerical results proved the effectiveness of the algorithm.
Algorithms
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Heart Valve Diseases
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diagnostic imaging
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Humans
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Likelihood Functions
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Mitral Valve
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diagnostic imaging
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pathology
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Patient Positioning
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Probability
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Ultrasonography
5.Analysis of the Relevance between t-BHP Induced Apoptosis and STATs mRNA Expression in HEI-OC1 Cells
Zihao ZENG ; Xuan LUO ; Yuheng YAO ; Junyi WANG
Journal of Audiology and Speech Pathology 2018;26(2):162-166
Objective To study the biological effects of (Janus Kinase-Signal transducers and activators of transcription,AK-STATs)path-way in hair cell apoptosis by analyzing the expression level of signal transducers and activators of transcription (STATS) of apoptotic HEI-OC1 cells line induced by oxidative stress injury.Methods Using different poisoning concentrations (0,20,40,60μM) of(Tert-Butyl hydroperoxide,T-BHP) built the apoptotic HEI-OC1 cells model while 0 μM was used as the control group.The apoptosis level at different poisoning concentration groups was detected by Annexin V-FITC/PI and the expression level of STATs mRNA by real-time quantitative PCR detecting system.Results The apoptosis rates in different contamination groups(0,20,40,60 μM) were 3.9%,7.4%,32.0%,and 91.2%,respectively.Compared with control group,STAT1 mRNA,STAT3 mRNA,STATS mRNA,and STAT6 mRNA expression levels in different poisoning concentration groups declined significantly (F=5 534.302,P<0.01;F=146.038,P<0.01;F=685.929,P<0.01;F=516.11,P< 0.01).No statistically significant differences were found among the poisoning concentration groups (P>0.05).Compared with the control group,the STAT2 mRNA expression levels changed significantly (F=1 259.148,P< 0.01).The STAT2 mRNA expression level in 20 μM contamination group decreased while 40,60 μM contamination group increased significantly (P<0.01).No expression of STAT4 in HEI-OC1 cell was noted.Conclusion The JAK-STAT2 signal pathway has the biological effects on leading oxidative stress injury apoptosis and JAK-STAT1/STAT3/STAT5/STAT6 has the biological effect of inhibit oxidative stress injury apoptosis.
6. The influence of STAT2 siRNA inhibition on tert-butyl hydroperoxide-induced apoptosis in HEI-OC1 cells
Shoukai LIN ; Zihao ZENG ; Min ZHANG ; Junyi WANG
China Occupational Medicine 2019;46(05):581-585
OBJECTIVE: To analyze the influence of signal transducers and activators of transcription 2(STAT2) on the apoptosis of oxidative stress-induced HEI-OC1 hair cells. METHODS: With random number table method,HEI-OC1 hair cells were divided into interference group and non-interference group. The artificial synthesis design targeted STAT2 small interfering RNA(siRNA) and opti-modified Eagle medium(opti-MEM) were added into the interference group, and only the opti-MEM was added into the non-interfere group. The 0, 20, 40, 60 μmol/L of tertiary butyl peroxide hydrogen(t-BHP) was added into the interference and non-interference group. Western blotting was used to detect the relative expression of STAT2 and cysteine protease(caspase)-3. The apoptotic rate of hair cells was detected by flow cytometry. RESULTS: In the interference group and non-interference group, the apoptotic rate of HEI-OC1 cells and the relative expression level of STAT2, Caspase-3 increased with the dose of t-BHP(P<0.01). The apoptotic rate of HEI-OC1 cells and the relative expression level of Caspase-3 in the interference group with 20, 40, 60 μmol/L of t-BHP were higher than that of the 20, 40 μmol/L group(P<0.01). The apoptotic rate of HEI-OC1 cells in the interference group was higher than those in the non-interference group with the same dose(P<0.01), while the relative expression level of STAT2 was lower than those in the non-interference group with the same dose(P<0.01). CONCLUSION: Inhibiting oxidative stress and STAT2 expression in hair cells can lead to aggravation of apoptosis. STAT2 has anti-apoptotic effect.
7.Establishment of prognostic scoring system for single large hepatocellular carcinoma after hepatectomy
Junyi SHEN ; Chuan LI ; Tianfu WEN ; Lvnan YAN ; Jiayin YANG ; Yong ZENG ; Hong WU ; Wentao WANG ; Mingqing XU ; Zheyu CHEN ; Yonggang WEI ; Li JIANG ; Jiwei HUANG
Chinese Journal of Hepatic Surgery(Electronic Edition) 2018;7(3):197-201
Objective To establish a clinical scoring system for the prognosis of patients with single large hepatocellular carcinoma (HCC) after hepatectomy.Methods 268 patients with single large HCC who underwent hepatectomy in West China Hospital of Sichuan University from January 2009 to December 2013 were included in this prospective study. There were 227 males and 41 females, of which 198 cases aged≤60 years old, 70 aged>60 years old. The informed consents of all patients were obtained and the local ethical committee approval was received. The patients' survival were observed. The independent risk factors for postoperative prognosis of patients with single large HCC were selected by Cox proportional risk regression mode. Based on the risk factors, the prognostic scoring system for single large HCC wasestablished. The scoring system was tested through survival analysis by Kaplan-Meier and Log-rank test. Results The median overall postoperative survival was 45 months, and the tumor-free survival was 31 months. Platelet-to-lymphocyte ratio (PLR)≥107, tumor diameter≥6.8 cm and positive microvascular invasion (MVI) were the independent risk factors for postoperative overall survival and tumor-free survival in patients with single large HCC (HR=1.004, 1.092, 2.233 and 1.003, 1.062, 1.534; P<0.05). Every independent risk factor was assigned 1 point. All patients were divided into low risk group (0 point), moderate risk group (1-2 points) and high risk group (3 points). The 5-year survival rate of high risk group was 25.4%, and that of moderate and low risk group was 33.2% and 52.1% respectively, where significant difference was observed (χ2=23.1, P<0.05). Similar Results were observed when the scoring system was used in patients with or without cirrhosis.Conclusions PLR≥107, tumor diameter≥6.8 cm and positive MVI are the independent risk factors for the prognosis of patients with single large HCC after resection. The prognostic scoring system established in this study can be used to predict the postoperative long-term survival of patients.
8.Efficacy of Lenvatinib Combined with Anti–PD-1 Antibodies Plus Transcatheter Arterial Chemoembolization for Hepatocellular Carcinoma with Portal Vein Tumor Thrombus: A Retrospective, Multicenter Study
Xiangye OU ; Junyi WU ; Jiayi WU ; Yangkai FU ; Zhenxin ZENG ; Shuqun LI ; Yinan LI ; Deyi LIU ; Han LI ; Bin LI ; Jianyin ZHOU ; Shaowu ZHUANG ; Shuqun CHENG ; Zhibo ZHANG ; Kai WANG ; Shuang QU ; Maolin YAN
Cancer Research and Treatment 2024;56(4):1207-1218
Purpose:
The prognosis of patients with hepatocellular carcinoma (HCC) and portal vein tumor thrombus (PVTT) is extremely poor, and systemic therapy is currently the mainstream treatment. This study aimed to assess the efficacy and safety of lenvatinib combined with anti–programmed cell death-1 antibodies and transcatheter arterial chemoembolization (triple therapy) in patients with HCC and PVTT.
Materials and Methods:
This retrospective multicenter study included patients with HCC and PVTT who received triple therapy, were aged between 18 and 75 years, classified as Child-Pugh class A or B, and had at least one measurable lesion. The overall survival (OS), progression-free survival (PFS), objective response rates, and disease control rates were analyzed to assess efficacy. Treatment-related adverse events were analyzed to assess safety profiles.
Results:
During a median follow-up of 11.23 months (range, 3.07 to 34.37 months), the median OS was greater than 24 months, and median PFS was 12.53 months. The 2-year OS rate was 54.9%. The objective response rate and disease control rate were 69.8% (74/106) and 84.0% (89/106), respectively; 20.8% (22/106) of the patients experienced grade 3/4 treatment-related adverse events and no treatment-related deaths occurred. The conversion rate to liver resection was 31.1% (33/106), with manageable postoperative complications. The median OS was not reached in the surgery group, but was 19.08 months in the non-surgery group. The median PFS in the surgery and non-surgery groups were 20.50 and 9.00 months, respectively.
Conclusion
Triple therapy showed promising survival benefits and high response rates in patients with HCC and PVTT, with manageable adverse effects.
9.A consensus on the management of allergy in kindergartens and primary schools
Chinese Journal of School Health 2023;44(2):167-172
Abstract
Allergic diseases can occur in all systems of the body, covering the whole life cycle, from children to adults and to old age, can be lifelong onset and even fatal in severe cases. Children account for the largest proportion of the victims of allergic disease, Children s allergies start from scratch, ranging from mild to severe, from less to more, from single to multiple systems and systemic performance, so the prevention and treatment of allergic diseases in children is of great importance, which can not only prevent high risk allergic conditions from developing into allergic diseases, but also further block the process of allergy. At present, there is no consensus on the management system of allergic children in kindergartens and primary schools. The "Consensus on Allergy Management and Prevention in Kindergartens and Primary Schools", which includes the organizational structure, system construction and management of allergic children, provides evidence informed recommendations for the long term comprehensive management of allergic children in kindergartens and primary schools, and provides a basis for the establishment of the prevention system for allergic children.
10.Therapeutic Effect of Sargentodoxae Caulis on Ulcerative Colitis and Exploring the Mechanism Based on GEO Chip Combined with Network Pharmacology
Feng XU ; Piao YU ; Linlin DU ; Qian ZENG ; Junyi WANG ; Hongmei WU ; Xiangpei WANG
Chinese Journal of Modern Applied Pharmacy 2024;41(3):332-340
OBJECTIVE
To study the anti-ulcerative colitis(UC) effect of Sargentodoxae Caulis and explore its mechanism.
METHODS
The UC mice model induced by dextran sodium sulfate was used to evaluate the anti-UC effect of Sargentodoxae Caulis. The ingredients of Sargentodoxae Caulis were obtained according to the CNKI and PubMed website, component targets were screened by SwissTargetPrediction database, GEO gene chip was used to extract UC differential genes, then a network of "ingredients-targets-disease" of the Sargentodoxae Caulis was constructed. After screening the core targets, protein interaction and cluster analysis, biological process and pathway enrichment analysis were performed, and the reliability of network analysis was preliminarily verified by molecular docking and literatures.
RESULTS
Sargentodoxae Caulis could significantly improve the disease activity index score, colon shortening and colonic histopathological changes of UC mice, and had a good anti-UC effect. The network analysis found that the core components of the anti-UC of Sargentodoxae Caulis include (+)-Dihydroxyurearetic acid, Isorhaponigenin and Pinosylvin, and 63 core targets, such as EGFR, STAT1 and LCK, regulating PI3K-Akt signal pathway and cancer proteoglycan and other related signal pathways of immune anti-inflammatory and anti-cancer, and it could affect the biological processes such as amino acid modification, kinase activity regulation, cell reaction and oxidative stress to treat UC. Molecular docking and literature showed that the constructed network had high reliability.
CONCLUSION
Sargentodoxae Caulis has a good anti-UC effect, and its mechanism may be closely related to the regulation of intestinal immune inflammation and cell proliferation, differentiation and migration. It has the characteristics of multi-component, multi-target and multi-way.