1.Study on differentially expressed genes associated with cell signal transduction between the intractable and non-intractable epilepsy rat brain by cDNA microarray
Yangmei CHEN ; Yunlan XIE ; Changlin HU ;
Chinese Journal of Laboratory Medicine 2001;0(03):-
Objective To explore the pathogenic mechanisms of intractable epilepsy by searching for the differential cell signal transduction associated genes expression in intractable and non intractable epilepsy rat brain using cDNA microarray Methods Intractable epilepsy and non intractable epilepsy rat model were build The total RNAs were isolated from the brain tissues Both mRNAs from the brains of the intractable and non intractable epilepsy rats were reversely transcribed to the cDNA with the incorporation of fluorescent dUTP to prepare the hybridization probes The PCR products of 4 096 human genes were spotted on a chemical material coated glass plates in array The mixed probes were then hybridized to the cDNA microarray After high stringent washing, the cDNA microarray was scanned for the fluorescent signals and showed differences between 2 tissues Results Among the 4 096 target genes, 29 genes associated with cell signal transduction differentially expressed were identified, 10 were up regulated(34 48%) and 19 down regulated(65 52%) Conclusion cDNA microarray technology is an effective technique in screening the differentially expressed genes between intractable and non intractable epilepsy rat brain Disturbances of cell signal transduction play a role in the pathogenic mechanism of intractable epilepsy
2.Expression and significance of signaling pathway of miR-134/CREB/pCREB in patients with epilepsy and in epileptic rats
Qian WANG ; Yangmei CHEN ; Jing GUO ; Xiaolan YANG ; Yunlan XIE
Acta Laboratorium Animalis Scientia Sinica 2014;(6):28-33
Objective To investigate the expression of microRNA-134 ( miR-134 ) , CREB and pCREB in the temporal lobe tissue of patients and epileptic rats and to explore their roles in pathogenesis of epilepsy.Methods Tempo-ral lobe tissue samples of 14 patients with refractory epilepsy and 10 non-epileptic patients, and hippocampus and brain tis-sue samples of 42 rats were used in this study.Forty-two healthy adult male Sprague-Dawley rats were randomly divided in-to 6 epilepsy groups (24 h, 72 h, 7 d, 14 d, 30 d, and 60 d after kindling epilepsy) and a normal control group (n=6 for all groups) .The rat model of epilepsy was generated by intraperitoneal injection of 127 mg/kg lithium chloride and 16-20 h later, 35 mg/kg pilocarpine.In the temporal lobe tissue of patients and hippocampal tissue of rats, the expression level of miR-134 was detected by real-time polymerase chain reaction.The expression levels of CREB and pCREB were de-termined by Western blot, and CREB and pCREB localization was assessed by immunohistochemistry.Results Compared with the control rats, the expression of miR-134 was significantly decreased in the temporal lobe tissue of experimental rats at 72 h,7 d,14 d, 60 d after kindling (P<0.05),and no significant change at 24 h and 30 d after kindling (P>0.05). Expression of miR-134 in patients with refractory epilepsy was significantly lower than that of the controls ( P<0.05 ) , while up-regulation of CREB expression was at the same time points (P<0.05).Up-regulation of pCREB expression was at all the time points after kindling (P<0.05).CREB and p-CREB expressions were seen in the nuclei of neurons, and significantly higher in patients with refractory epilepsy and epileptic rats.Conclusions The expression of miR-134 is sig-nificantly decreased and that of CREB and pCREB was significantly increased in the temporal lobe tissue of patients with re-fractory epilepsy and the hippocampal tissue of epileptic rats.These findings indicate that the signaling pathway of miR-134/CREB/pCREB may play an important role in the pathogenesis of epilepsy.
3.SIRT1 expression and activity are up-regulated in the brain tissue of epileptic patients and rat models.
Yongping CHEN ; Yunlan XIE ; Heng WANG ; Yangmei CHEN
Journal of Southern Medical University 2013;33(4):528-532
OBJECTIVETo investigate the expression and activity of silent information regulator 1 (SIRT1) in the temporal lobe of epileptic patients and rat models and explore its role in the occurrence and progression of epilepsy.
METHODSThe temporal lobe tissue of epileptic patients and rat models (induced by lithium-pilocarpine) were examined for SIRT1 expression using immunohistochemistry and Western blotting and also for SIRT1 activity using SIRT1 Deacetylase Assay Kit.
RESULTSImmunohistochemistry detected positive SIRT1 expression mainly in the cytoplasm of the neurons in both human and rat brains, and the epileptic groups showed stronger SIRT1 immunoreactivity than the control group. Western blotting and activity assay showed that the expression and activity of SIRT1 were significantly increased in the temporal lobe of patients with refractory epilepsy as compared with the tissues samples from non-epileptic patients (P<0.05). In the rat models of epilepsy, SIRT1 expression was up-regulated at 6, 24, and 72 h and at 7, 14, 30, and 60 days after kindling (P<0.05) and SIRT1 activity was significantly increased at 6, 24, and 72 h and at 7 and 14 days (P<0.05), with the peak level of SIRT1 expression and activity occurring at 72 h.
CONCLUSIONUp-regulation of SIRT1 expression and activity in the temporal lobe of epileptic patients and rat models may play an important role in the pathogenesis of epilepsy.
Adolescent ; Adult ; Animals ; Disease Models, Animal ; Epilepsy ; metabolism ; Female ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Sirtuin 1 ; metabolism ; Temporal Lobe ; metabolism ; Young Adult
4.Clinical value of different samples of EBV-DNA detection in patients with infectious mononucleosis
Shizhi CHEN ; Yunlan XIE ; Yanhua MEI ; Zhijia ZHUANG
International Journal of Laboratory Medicine 2018;39(2):176-178,181
Objective To investigate the diagnostic value of plasma ,peripheral blood mononuclear cell (PB-MC) and throat swab EB virus (SBV) load in the patients with infectious mononucleosis (IM ) aged over 16 years old .Methods The detection results in 130 patients with suspected IM aged over 16 years old of 3 differ-ent samples of plasma ,peripheral blood mononuclear cell(PBMC) and throat swab EB virus(SBV) load were analyzed retrospectively .Results Among 61 cases of IM verified by clinic and laboratory ,16 cases of IM were detected by plasma sample ,the detection rate of EBV-DNA was 26 .22% ;in 31 cases of PBMC detection ,the EBV-DNA detection rate was 61 .29% ;in 31 cases of throat swab detection ,the EBV-DNA detection rate was 83 .78% .The EBV-DNA detection rate of the combined detection with one indicator or multiple indicators positive of plasma ,PBMC and throat swab as the basis was 100 .00% .Conclusion The combined detection of plasma ,PBMC and throat swab can improve the positive rate in the patients with IM aged over 16 years old , w hich can provide more bases for pathogenic diagnosis and treatment for clinic .
5.Intraocular pressure distribution and reference interval of high-altitude eye health screening population from Xining, Qinghai
Yiquan YANG ; Yunlan BAO ; Yunxiao SUN ; Yuan XIE ; Xialing WANG ; You LIU ; Yanling XIE ; Jie HAO ; Xiaoxia PENG ; Sujie FAN ; Shizheng WU ; Ningli WANG
Chinese Journal of Experimental Ophthalmology 2023;41(2):166-172
Objective:To investigate the distribution of intraocular pressure (IOP) in high-altitude population aged 18 years and over in Xining, Qinghai and establish the reference interval (RI) of IOP.Methods:A cross-sectional study was conducted in Xining, Qinghai Province at 2.271 km above sea level from September 2019 to May 2020.Ophthalmic examinations and IOP measurement were conducted among subjects from Physical Examination Center of Qinghai Provincial People's Hospital.The subjects who had been living in Xining without leaving for three months were enrolled.Ophthalmic examinations included vision examination, IOP measurement, slit-lamp microscopy, fundus photography, anterior and posterior segment optical coherence tomography.IOP was measured using Goldmann applanation tonometry under local anesthesia.Subjects with factors that could cause significant changes in IOP and affect the accuracy of IOP measurement, and those who were unable to receive IOP measurement were excluded.Subjects were grouped according to sex, age and ethnicity, and the distribution and RI of IOP were compared among all groups.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Beijing Tongren Hospital, Capital Medical University (No.TRECKY2017-024). Written informed consent was obtained from each subject.Results:A total of 6 120 subjects (6 120 eyes) aged 18-90 years old were enrolled, including 2 850 males and 3 270 females with average age of (45.54±13.85) years.The average IOP of high-altitude population in Xining, Qinghai Province was (14.32±1.93) mmHg (1 mmHg=0.133 kPa), with the RI of 10.54-18.10 mmHg.The average IOP was (14.42±1.98) mmHg in male with the RI of 10.54-18.30 mmHg, (14.23±1.88) mmHg in female with the RI of 10.55-17.91 mmHg.The IOP of male was higher than that of female ( t=3.71, P<0.001). The IOP of Han, Tibetan, Hui and other nationalities were (14.38±1.91), (13.93±2.06), (14.21±1.87), (13.94±1.95) mmHg, respectively, with a statistically significant overall difference ( F=6.73, P<0.001). The IOP of Han nationality was significantly higher than that of Tibetan, Hui and other nationalities, and the differences were statistically significant (all at P<0.05). Conclusions:RI of IOP in high-altitude population from Xining, Qinghai is lower compared with normal altitude area.