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.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.
8.Research progress on vascularization of organoids.
Junyi SHEN ; Zhi OUYANG ; Jian ZHONG ; Yicen LONG ; Yujia SUN ; Ye ZENG
Journal of Biomedical Engineering 2023;40(4):625-631
Organoids are three-dimensional structures formed by self-organizing growth of cells in vitro, which own many structures and functions similar with those of corresponding in vivo organs. Although the organoid culture technologies are rapidly developed and the original cells are abundant, the organoid cultured by current technologies are rather different with the real organs, which limits their application. The major challenges of organoid cultures are the immature tissue structure and restricted growth, both of which are caused by poor functional vasculature. Therefore, how to develop the vascularization of organoids has become an urgent problem. We presently reviewed the progresses on the original cells of organoids and the current methods to develop organoids vascularization, which provide clues to solve the above-mentioned problems.
Humans
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Organoids
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Neovascularization, Pathologic
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Technology
9. Comparative analysis on inter-laboratory detection of free silica indust in Guangdong Province,2012-2015
Yi SUN ; Junyi HUANG ; Aihua ZHANG ; Banghua WU ; Zhengnong HUANG ; Weihui WANG ; Xiaozhong YANG ; Feifei ZENG
China Occupational Medicine 2017;44(02):207-210
OBJECTIVE: To evaluate the technical level and ability of the detection of free silica in dust in occupational health technical service institutions of Guangdong Province through inter-laboratory comparison. METHODS: The interlaboratory detection of free silica in dust in 55,61,58 and 66 occupational health technical service institutions were carried out and compared in 2012-2015. The results were statistically analyzed by four points robust statistical technique,and the relative deviation between the laboratory test results and the Median of them was used as the auxiliary evaluation index of the laboratory's technical ability. RESULTS: The stable coefficient of variation(CV) of samples with free silica content > 20. 00% was less than 5. 00%,and the stable CV of samples with free silica content ≤20. 00% was more than5. 00%. The qualified rate in 2012-2015 were 89. 09%,81. 97%,77. 59% and 81. 82%,respectively. There was no statistical difference(P > 0. 05) in the annual reference between private and non-private institutions compared to the qualified rate in 2012-2015. There was no statistical difference(P > 0. 05) in the qualified rate of the institutions in the Pearl River Delta region and in the non-triangular regions of the Pearl River Delta in 2012-2015. CONCLUSION: Comparison of the detection technology service ability in the detection of free silica in dust should be conducted to improve its technical service quality.
10.Association between wrist pain and awkward postures among workers in 10 key industries
Guanlin LI ; Xin SUN ; Meibian ZHANG ; Huadong ZHANG ; Ruijie LING ; Yimin LIU ; Gang LI ; Nengzhou CHEN ; Zaoliang REN ; Yan YIN ; Hua SHAO ; Hengdong ZHANG ; Jiajie LI ; Bing QIU ; Dayu WANG ; Qiang ZENG ; Zhanhui LIANG ; Rugang WANG ; Jianchao CHEN ; Danying ZHANG ; Liangying MEI ; Yongquan LIU ; Jixiang LIU ; Chengyun ZHANG ; Tianlai LI ; Ning JIA ; Junyi WANG ; Zhongxu WANG ; Qingsong CHEN
Journal of Environmental and Occupational Medicine 2023;40(1):49-54
Background Prolonged awkward postures during occupational activities can lead to excessive musculoskeletal load on the wrist of workers and symptoms such as wrist pain or discomfort. Objective To survey the prevalence of wrist pain among workers in 10 key industries and analyze its correlation with wrist working postures. Methods By using stratified cluster sampling method, workers from 10 key industries, such as footwear manufacturing industry, shipbuilding manufacturing industry, and automobile manufacturing industry, were selected from seven regions in North China, East China, Central China, South China, Southwest China, Northwest China, and Northeast China. The demographic information, wrist working postures, pain in wrist of the workers were collected through a cross-sectional survey. Pearson χ2 test was used to compare prevalence by selected factors, trend χ2 test for between group comparison, and unconditional logistic regression models for the association of wrist working postures with wrist pain. Results There were 64052 workers enrolled in this survey, and 56286 provided valid questionnaires (the effective rate was 87.8%). According to the survey, the prevalence of wrist pain was 23.3% (13112/56286), and the industries with higher prevalences were footwear manufacturing (27.1%, 1927/7106), automobile manufacturing (24.9%, 5378/21560), and shipbuilding and related equipment manufacturing (24.4%, 850/3488) industries. Finger pinching (OR=2.09, 95%CI: 1.95-2.24), frequent wrist bending (OR=2.03, 95%CI: 1.92-2.15), fixed wrist bending (OR=1.77, 95%CI: 1.69-1.85), wrist on hard edge (OR=1.34, 95%CI: 1.28-1.40), and arms over shoulders (OR=1.11, 95%CI: 1.05-1.17) increased the risk of reporting wrist pain. Conclusion Awkward postures are related to wrist pain among workers in selected 10 key industries. The related factors are wrist on hard edge, frequent wrist bending, finger pinching, fixed wrist bending, and arms over shoulders.