1.Expression Profile and Bioinformatics Studies of Long Noncoding RNA in Mouse Embryonic Fibroblasts under Endoplasmic Reticulum
Hongyang QUAN ; Qianqian FAN ; Lin WANG
Journal of Medical Research 2015;44(4):61-66
Objective To explore the potential significance of long noncoding RNA (lncRNA) expressed in endoplasmic reticulum stress response also known as unfolded protein response ( UPR) .Methods Mouse embryonic fibroblasts ( MEFs) were isolated from 13 day mouse embryo , cultured and treated with tunicamycin for 16 hours to induce UPR .The untreated MEFs were used as negative control .The lncRNA expression profile was examined by a customized lncRNA array. The results of array assay were validated by real-time PCR and further analyzed by bioinformatics. Results There were 411 lncRNAs whose expression was signficantly up-regulated in MEFs treated with tunicamycin as compared with the controls ,while 790 lncRNAs were significantly down-regulated. A number of significantly alter lncRNAs were validated by real-time PCR. A further bioinformatics analysis of lncRNA profile suggested that many cold be in-volved in multiple cellular pathways. Conclusion Our lncRNA expression profile and bioinformatics studies srongly suggest that many lncRNAs can be regulated in the cells under UPR and could also in turn regulate UPR and other cellular processes.
2.The study of application of step-wise diversified teaching mode in cardiac electrophysiology
Hongyang GUO ; Kun LIN ; Ye LI ; Zhaoliang SHAN ; Yutang WANG
Chinese Journal of Medical Education Research 2014;(7):712-715
Objective To investigate the effect of a step-wise diversified teaching mode on cardiac electrophysiology (EP) education. Methods 64 young doctors (male:36, female:28) who studied in EP sub-specialty were enrolled. The atrioventricular nodal reentrant tachycardia (AVNRT) and atrioventricular reentrant tachycardia (AVRT) were selected as teaching content. All the doctors were randomly divided into traditional teaching group (TT) and step-wise diversified teaching group (DT) according to cross-over design protocol. In TT group, only teaching and answering the questions by teachers were given to students, for the DT group, in different stages of teaching, multiple teach-ing strategy were combined to complete teaching. T test and Chi square test were used to compare the effect of two teaching methods. Results There was no difference of basic features and time of self-studying between the two groups. In examinations, the students in DT group got higher score than TT group[DT:(92.3±9.8) and (93.1±7.8),TT:(88.3±8.6) and (87.1±10.0),P<0.05], and had better perfor-mance of learning interest, initiative, ability of finding and resolving problems, teamwork spirit and so on. Conclusion The step-wise diversified teaching mode contributes improving the quality of instruction of cardiac EP, moreover, arousing learning interest and initiative, enhancing ability of find-ing and resolving problems, teamwork spirit and so on, which is worthy of wide-popularizing in cardiac EP training.
4.PPM1D silencing by lentiviral-mediated RNA interference inhibits proliferation and invasion of human glioma cells.
Peng, WANG ; Jing, RAO ; Haifeng, YANG ; Hongyang, ZHAO ; Lin, YANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(1):94-9
To construct a lentiviral shRNA vector targeting human protein phosphatase 1D magnesium-dependent (PPM1D) gene and detect its effectiveness of gene silencing in human gliomas, specific siRNA targets with short hairpin frame were designed and synthesized. DNA oligo was cloned into the pFU-GW-iRNA lentiviral expression vector, and then PCR and sequencing analyses were conducted to verify the constructs. After the verified plasmids were transfected into 293T cells, the lentivirus was produced and the titer of virus was determined. Real-time quantitative PCR and Western blot were performed to detect the PPM1D expression level in the infected glioma cells. PCR and Western blot analyses revealed the optimal interfering target, and the virus with a titer of 6×10(8) TU/mL was successfully packaged. The PPM1D expression in human glioma cells was knocked down at both mRNA and protein levels by virus infection. The expression of PPM1D mRNA and protein was decreased by 76.3% and 87.0% respectively as compared with control group. The multiple functions of human glioma cells after PPM1D RNA interference were detected by flow cytometry and cell counting kit-8 (CCK-8). Efficient down-regulation of PPM1D resulted in significantly increased cell apoptosis and reduced cell proliferation and invasion potential in U87-MG cells. We have successfully constructed the lentiviral shRNA expression vector capable of stable PPM1D gene silencing at both mRNA and protein levels in glioma cells. And our data gave evidence that the reduced cell growth observed after PPM1D silencing in glioma cells was at least partly due to increased apoptotic cell death.
5.Magnetic resonance imaging of transplanted neural stem cells in Parkinson disease rats.
Lin, YANG ; Ying, XIA ; Hongyang, ZHAO ; Jiashan, ZHAO ; Xianli, ZHU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2006;26(4):489-92
In this study we implanted magnetically labeled neural stem cells (NSCs) in PD rats and then monitored their survival and migration in the host brain by magnetic resonance imaging (MRI). The mesencephalic NSCs were obtained from the brain of SD rats. Superparamagnetic iron oxide (SPIO) was transferred to NSCs by Lipofectamine transfection. Eighteen PD lesioned rats were selected for transplantation by evaluation of their rotational behavior in response to amphetamine and randomly assigned to 3 groups, i.e., sham group, PBS group and NSCs transplanted group, with 6 rats in each group. MR scanning was performed at 1, 2, 4, 6, 8 and 10 week(s) following transplantation. At the meantime, rotational behavior was assessed in each group. Our results showed that SPIO particles were clearly visible with Prissian blue staining in neurospheres and cells derived from NSCs. The rotational behavior of the NSCs transplanted group was remarkably improved compared with that of sham group and PBS group (P < 0.05). In vivo MR tracking of NSCs showed that SPIO labeling led to a strong susceptibility change of signal 1 week after transplantation on T2 weighted images. And a large circular hypointense signal appeared in the transplanted area on T2* gradient echo images. Ten weeks following transplantation, the hypointense signal on T2 weighted and T2* gradient echo images was still displayed. It is concluded that SPIO particles could label NSCs effectively, and MRI detection of SPIO labeled cells is a promising method and novel approach to analyzing the NSCs following transplantation in the treatment of PD.
6.The Follow-up Study for Clinical Compliance in Patients With Permanent Pacemaker Implantation
Yuexiang ZHAO ; Zhaoliang SHAN ; Hongyang GUO ; Kun LIN ; Jianping GUO ; Yutang WANG
Chinese Circulation Journal 2014;(10):784-786
Objective: To analyze the current condition and inlfuencing factors for clinical compliance in patients with permanent pacemaker implantation and to improve the follow-up condition in relevant patients. Methods: A total of 817 patients with permanent pacemaker implantation in our hospital from 2006-01 to 2013-01 were retrospectively studied. The clinical compliance condition and inlfuencing factors were accessed for 1 year period. Results: There were 26/817 (3.18%) patients lost contact and 1 patient died. A total of 790 (96.7%) patients finished the followed-up study by 2 groups: Clinical visit group,n=440 (55.70%) and Telephone visit group,n=350 (44.30%). The education level, medical cost, residency, comprehension of arrhythmia and accompany personnel were different between 2 groups,P<0.05. The patients were with high school education or above, reimbursed medical cost, local residency, comprehension of arrhythmia and accompany personnel had the higher clinical visit rate. The overall 1 year occurrence rate of complication was 1.8% without severe event. There were 59.5% of patients optimized the pacemaker parameters during clinical visit. Conclusion: The patients with permanent pacemaker implantation had the lower rate of clinical follow-up visit which should be improved in several issues.
7.Impact of body mass index on the development of pocket hematoma:A retrospective study in Chinese people
Jianping GUO ; Zhaoliang SHAN ; Hongyang GUO ; Hongtao YUAN ; Kun LIN ; Yuexiang ZHAO ; Yutang WANG
Journal of Geriatric Cardiology 2014;(3):212-217
BackgroundPocket hematoma is one of the major complications associated with cardiovascular implantable electronic devices (CIEDs) implantation. The aim of this study is to evaluate the impact of body mass index (BMI) on the occurrence of pocket hematoma after CIEDs implantation.MethodsThe study is a retrospective review of 972 patients receiving CIEDs implantation between 2008 and 2012 in a tertiary hospital.ResultsTwenty two patients (2.2%) developed severe pocket hematoma requiring re-intervention. The hematoma rate (4.6%,n = 15) of patients with a BMI of < 23 kg/m2 was significantly higher compared with that of patients with a BMI of≥23 kg/m2 (1.1%, n = 7,P< 0.001). In multivariate regression analysis, a BMI < 23.0 kg/m2 may be associated with the development of severe pocket hema-toma. An increase of 1.0 kg/m2 in BMI was associated with lower incidence of hematoma formation (OR: 0.84; 95% CI: 0.74-0.95;P = 0.006).ConclusionBMI < 23 kg/m2 was associated with a higher incidence of pocket hematoma, requiring re-intervention. The data sup-port that great care must be taken when patients were with a lower BMI received CIEDs implantation.
8.Expression of neuron-specific enolase protein in rats after intermittent hypoxia and its influence on learining and memory function
Panpan ZHANG ; Xiaoqing HAN ; Hongyang WANG ; Jiangtao YU ; Lin LI ; Limin WANG ; Xiuhua GUO
Chinese Journal of Behavioral Medicine and Brain Science 2014;23(4):303-306
Objective To observe the changes of learning function and neuron-specific enolase (NSE)protein of both brian and serum in rats after intermittent hypoxia,and explore the relation between NSE protein and learning and memory function.Methods Male Wistar adult rats(n=48)were randomly divided into control group (UC group) and 7.5% chronic intermittent hypoxia group (7.5%CIH group).7.5% chronic intermittent hypoxia model in rats were simalated by using the self-made cabin of intermittent hypoxia.At the 2nd,4th,6th,and 8th week respectively,learning and memory function in rat was tested by Morris water maze in two groups.The level of NSE protein in hippocampal CA1 region was detected by immunohistochemical method,and was detected in serum of rats by using enzyme-linked immunosorbent adsorption method.Results Compared with the control group,7.5% CIH group at the 2nd,4th,6th,and 8th week in rats from the second week of the escape latency time was 44.13± 2.84) s,(50.35 ± 1.96) s,(57.47 ± 1.66) s,(62.85 ± 1.80) s,and across the target quadrant time was 48.81 ± 2.09) s,(42.04± 1.84) s,(36.82± 2.07) s,(31.81 ± 1.68) s.From the first two weeks,the longer the hypoxia time prolonged,the longer the rat's escapedlatency (P<0.05) and the shorter the rats acrossed the target quadrant (P <0.05).7.5% CIH group of hippocampal CA1 neurons NSE protein at the 2nd,4th,6th,and 8th week was (9.69±1.37),17.10± 1.87),(24.79± 3.51),(34.16±5.35),respectively,and serum NSE protein at the 2nd,4th,6th,and 8th week respectively was (6.03±0.91) μg/L,(11.04± 1.89) μg/L,(16.39± 1.00) μg/L,(24.22±3.73) μg/L,both of which were more than the control group.The difference was statistically significant (P<0.05).7.5% CIH group of hippocampal CA1 neurons and serum NSE protein expression were significant time differences,and gradually increased over time.The difference was statistically significant (P<0.05).Positive NSE protein expression in hippocampal CA1 region relative NSE and serum protein expression values were positively correlated (r=0.94,P <0.01).But in control group NSE protein expression did not exist time difference (P>0.05).Conclusion Intermittent hypoxia can cause NSE levels significant increase in serum and brain tissue,and the dynamic changes of them can reflect severity of learning and memory impairement in rat.
9.Effect of continuous positive airway pressure ventilation on serum levels of s100β protein and neuron-specifice enolase in obstructive sleep apnea-hypopnea syndrome patients.
Panpan ZHANG ; Xiaoqing HAN ; Hongyang WANG ; Lin LI ; Ling WANG ; Min ZHANG ; Chen LIU ; Jiangtao YU
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2015;29(6):509-512
OBJECTIVE:
To explore the effect and clinical significance of serum S100β and NSE on moderate and severe obstructive sleep apnea syndrome (OSAHS) after the continuous positive airway pressure (CPAP).
METHOD:
A total of 60 cases with obstructive sleep apnea were choosed with PSG in our hospital in June 2009 to June 2009. According to apnea hypoventilation index and at night the lowest oxygen saturation, divided into severe group (n=60) and moderate group (n=60), selecting 60 cases of healthy physical examination for a control over the same period. According to the length of the course of the disease in patients with each group can be divided into <5 years group, 5-10 years and > 10 years group, severe and moderate groups were recruited to undergo an CPAP treatment,both before and after treatment for 3 months, the lowest oxygen saturation, average blood oxygen saturation and apnea hypoventilation index were determined in moderate and severe groups with PSG, at the same time, serum S100β and NSE were determined with immune enzyme-linked adsorption testing before and after patients in different course of treatment and control group.
RESULT:
Compared with pretherapy of severe and moderate groups, the lowest oxygen saturation, average blood oxygen saturation and apnea hypoventilation index were ower after treatment (P<0. 05), serum S100β and NSE in severe and moderate groups before and after treatment were significantly higher than control group (P<0. 05), and two groups > 10 years before and after treatment in patients with serum according to beta and NSE levels higher than 5-10 patients, 5-10 patients before and after treatment according to beta and NSE serum levels higher than <5 years group of patients, the relation between the two groups of patients before and after treatment according to beta and NSE serum levels with the extension of history time increased. Before the treatment serum according to beta and NSE in patients with severe group were higher than moderate group before treatment (P< 0.05). Relation between the two groups after treatment according to beta and serum NSE was significantly decreased the (P<0. 05), the relation between the two groups after treatment according to beta and serum NSE, there was no statistically significant difference (p>0. 05), the relation between two groups according to beta, NSE serum are positively correlated with AHI (P < 0. 01).
CONCLUTION
CPAP significantly reduced serum S100β and NSE levels in patients with OSAHS, both may be important index which evaluated nervous system protection of CPAP in patients with OSAHS.
Continuous Positive Airway Pressure
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Humans
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Phosphopyruvate Hydratase
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blood
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S100 Calcium Binding Protein beta Subunit
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blood
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Sleep Apnea, Obstructive
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blood
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therapy
10.Cloning of the Eukaryotic Expression Vector with Nerve Growth Factor in Rats and Its Effects on Proliferation and Differentiation of Mesencephal Neural Stem Cells of Fetal Rats
LIN MINHUA ; YANG LIN ; FU RONG ; ZHAO HONGYANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2008;28(5):513-516
Summary: The eukaryotic expression vector containing full-length cDNA sequence of rate nerve growth factor (NGF) β subunit was constructed and its effects on proliferation and differentiation of neural stem cells were observed. By using PCR, full-length cDNA sequence of NGF β subunit in rats was cloned and ligated into the eukaryotic expression vector pEGFP-N1-NGE The recombinant plasmid pEGFP-N1-NGF was transfected into the mesencephal neural stem cells of embryonic rats by Lipofectamin and transiently expressed. MTT method was used to determine the effects of NGF on proliferation of neural stem cells, and under phase-contrast microscopy, the effects of NGF on growth of nervous processes following differentiation of neural stem cells were observed. Sequence analysis indicated that the cloned full-length cDNA sequence of rat NGF β was identical to that of published sequence encoding NGF in gene GeneBank. The transfection of recombinant plasmid pEGFP-N1-NGF into mesencephal neural stem cells of embryonic rats could obviously promote proliferation of neural stem cells and faciliate the growth of neural stem cells-derived nerve cells. It was suggested that neural stem cells could be used as a vehicle of gene transfer, and the expression of NGF β subunit in the neural stem cells could promote the growth of nerve cells derived from neural stem cells.