1.Developmental rules of auditory event-related potentials of exogenous components:from children to adults
Xiaoyi CHEN ; Jie CAO ; Fangcheng CAI
Journal of Third Military Medical University 2017;39(17):1762-1767
		                        		
		                        			
		                        			Objective To investigate the normal waveform,lantency and amplitude of auditory event-related potentials (ERPs) of exogenous components in children at different ages,and summarize the developmental rules and clinical perspectives of exogenous ERPs according to the International Federation of Clinical Neurophysiology (IFCN) Recommended Standards in 2009.Methods One hundred and thirty-six children aged from 3 to 17 years old (admitted in the departments from healthcare,orthopedics,general surgery,and respiratory and digestive diseases,and those recovered from respiratory infection and gastrointestinal infection),and healthy adults (from undergraduates of Chongqing medical university) were recruited in this study from 2008 to 2010.They were divided into 4 groups in accordance with their ages,that is,3 ~5,> 5 ~7,> 7 ~9,and > 9 ~17 years old groups,and adult group (22 ~29 years old).ERPs were recorded from the midline site (Cz) with classical auditory Oddball paradigm.Each subject was tested twice or more to determine waveform repeatability.The overlapped graphs of ERPs under target and standard stimuli were obtained in the potentiometer.The exogenous ERPs,including P1,N1,P2 and N2,lantency and amplitude were measured.Results The latencies of exogenous ERPs under target and standard stimuli were shortened in a non-uniform manner with the increasing of age.No matter under target or standard stimuli,over 95% of mean latencies of exogenous ERPs were within the range of 2SD.In addition to N1 component,the amplitude of P1,P2 and N2 components induced by target stimulus were in a trend of increased first and then decreased with the increasing of age.Compared with the children,the amplitude was much lower in the adults and adolescents (P < 0.05).The N2 component induced by target stimulus was absent among 95% healthy adults.The Standard deviations of amplitudes of exogenous components were great in all aged groups,indicating significant individual differences.Conclusion The latencies of exogenous ERPs are shortened with the increasing of age.Target stimulus will produce more stable results than the standard stimulus,and the ERPs under target stimulus can reflect the primary auditory cortex function of temporal lobe,and are independent on patients' cooperation.
		                        		
		                        		
		                        		
		                        	
2.Developing an animal model of the brain stem myoclonus
Zhijiang HE ; Jie CAO ; Fangcheng CAI ; Chenggong FENG ; Hengsheng CHEN
Chinese Journal of Neurology 2010;43(1):39-44
		                        		
		                        			
		                        			Objective To develop experimental animal model of the brain stem myoclonus,which more closely replicate clinic features of mechanism, behavior, neuroelectrophysiology and pharmacodynamics.Methods L-5-HTP (the precursor of L-5-HT)was microinjected into the dorsal pons of young guinea pig to induce myoclonus (electromyogram burst of myoclonus≤400 ms by synchronous recording).Some animals were pretreated with anticonvulsant VPA,CZP or CBZ at effective dose 50 (EC_(50)).Myoclonus was induced when the drug level was within their effective anticonvulsion concentration.The neuroelectrophysiological characteristics of myoclonus including latency,time of reaching its peak,duration of seizure peak,the maximum seizure frequency and total duration were detected.EMG and ictal electroencephalogram(EEG)were recorded synchronously.The origin of myoclonus and its correlation with epileptic discharges were further confirmed by jerk-locked back averaging(JLA).Results (1)L-5-HTP induced pure myoclonus from the dorsal pons of guinea pig permanently(8/every site,the rate of producing myoclonus is 100%).(2)The myoclonus presented bilaterally or as general myoclonus,which was sensitive to tactile and sound sensation.(3)The EMG duration of the myoclonus wag longer((208.75 ± 81.42)ms),and ictal EEG showed scattered and irregular spikes and sharp waves without time-locked correlation with EMG activities.(4)The synchronous ictal EEG of the myoclonus showed spike and sharp waves,but there was no time-locked EEG activity in JLA.(5)In the animals treated with anticonvulsant at EC_(50) concentrations:VPA and CZP decreased the maximum seizure frequency(there are 28.13±3.79 per minutes in VPA group and 37.17±4.67 perminutes in CZP group)and shortened the duration of peak time ((55.00±14.14)minutes in VPA group and(50.00±11.73)minutes in CZP group respectively)and total time(VPA group was(124.17±40.04)minutes and CZP group was(156.88±30.71)minutes)of myoclonus(F value were between 23.41 and 35.44,P<0.01 or P<0.05).CBZ increased duration of peak time((98.75±13.86)minutes)and total time((257.50±14.79)minutes)of myoclonus(P<0.05 and 0.01).Conclusions The new model generates pure myoclonus originating from brain stem and also has a shorter duration of muscle construction(≤400 ms)and more sensitivity to tactile and sound sensation.Therefore,the model presents characteristics closer to the brain stem myoclonus in the clinic phenotype in respect of seizure behavior,pharmacodynamics and neuroelectrophysiology.
		                        		
		                        		
		                        		
		                        	
3.Pathogenicity of a mutant from Campylobacter jejuni O∶19 by deleting neuB1
Shuli XIANG ; Fangcheng CAI ; Xiaoping ZHANG
Journal of Third Military Medical University 2003;0(23):-
		                        		
		                        			
		                        			Objective To explore the pathogenicity of Campylobacter jejuni(CJ)O∶19 strain after deletion of neuB1.Methods The mutant of CJ O∶19 by deleting neuB1,which LPS is deficient in sialic acid,was compared with the homologous wild strain by assaying their sensitivity to bactericidal activity,motility and autoagglutination of 10% normal human serum(NHS).Results The sensitivity of the mutant strain to bactericidal activity of serum was superior to that of the wild strain.The survival rate of the mutant strain was(20.6?7.4)% and(9.6?3.6)% after incubation with 10% NHS for 15 min and 60 min,that of wild strain was(36.9?5.9)% and(15.5?4.3)% respectively.The wild and mutant strains showed no significant difference in sensitivity to motility and autoagglutination of NHS.Conclusion The mutant strain possesses remarkable pathogenicity so that an inactivated vaccine from the mutant strain should be first considered.
		                        		
		                        		
		                        		
		                        	
4.Antioxidants protects against peripheral nerve damages induced by antiepileptic drugs in rats
Min ZHONG ; Fangcheng CAI ; Xiaoping ZHANG ; Yan SONG
Journal of Third Military Medical University 2003;0(23):-
		                        		
		                        			
		                        			Objective To explore the protective efficacy of antioxidants (vitamin E and Ginkgo biloba extract) on the peripheral nerve damages induced by antiepileptic drugs (AEDs). Methods Adult (2-month old) and infant SD rats (7-day old) were respectively treated with AEDs (phenytoin,or phenobarbital,or clonazepam) alone,or simultaneously with vitamin E or Ginkgo biloba extract for 4 weeks. All the rats were sacrificed,sciatic nerves and serum were collected. The sciatic nerves were analyzed by histologically for their pathological changes,and serum and homogenate of sciatic nerves were investigated for their total antioxidative capacity and antioxidant enzyme activity. Results Incidence of pathological abnormalities of teased fibers significantly reduced in all rats treated with AEDs and antioxidants (P
		                        		
		                        		
		                        		
		                        	
5.Pathogenesis of brain damage in infant rats induced by corticosteroids
Xiaoyu WANG ; Jie CAO ; Fangcheng CAI
Journal of Third Military Medical University 2003;0(24):-
		                        		
		                        			
		                        			Objective To explore the possible pathogenesis of brain damage in rats induced by prednisone or corticotrophin. Methods The doses of prednisone or corticotrophin were determined by a primary experiment to obtain corresponding plasma cortisol or corticosterone level as same as that in sick children after drug taking. Then 192 healthy infant (at the age of 7 d) and 192 adult (at the age of 2 months) male SD rats were divided into 4 groups as infant prednisone group, adult prednisone group, infant corticotrophin group and adult corticotrophin group (n=96 in each). then every group was further subdivided into 12 subgroups (n=8 in each). The subgroups were divided according to the dose (therapeutic or low doses), the course [short (10 d) or long (3 weeks)], the sacrificed time (24 h or 4 weeks after withdrawal), and their corresponding controls. Serum neuron-specific enolase (NSE) concentration was quantified by ELISA. Expressions of apoptosis-related proteins, Bax and Bcl-2 in the brain were detected by immunohistochemical assays. Neuronal apoptosis was detected by TUNEL. Results Our primary experiment indicated that the therapeutic dose was 4 mg?kg-1?d-1 or 150 U?m-2?d-1 for prednisone or corticotrophin, and the small dose was 2 mg?kg-1?d-1 or 38 U?m-2?d-1 for them. In infant rats treated with prednisone or corticotrophin at therapeutic-dose for short or long term, their serum NSE concentration were increased significantly by 50.6% to 103.2%. And serum NSE was increased by 38.3% to 60.3% in infant after low-dose treatment for long term. Over-expression of Bax protein (P
		                        		
		                        		
		                        		
		                        	
6.Experimental study on susceptibility of high risk factors related with hepatotoxicity of valproic acid
Dagan FU ; Fangcheng CAI ; Xiaoping ZHANG
Chinese Pharmacological Bulletin 2003;0(11):-
		                        		
		                        			
		                        			Aim To explore the differences of pathogenesis for the hepatotoxicity induced by chronic treatment of valproic acid(VPA) in different ages,and in combination administration with inducers of liver enzyme.Methods Animal models were established by oral administration chronically with VPA at doses of 200 or 500 mg?kg~(-1) per day in 30 days for 50 Wistar rats(infant and adult rats) with inducers of liver enzyme Phenobarbital(PB) or not.Mitochondria were obtained by differential centrifugation.Levels of liver enzymes,coagulation factors,plasma ammonia,VPA and PB serum levels,and L-carnitine in sera,as well as the changes of respiratory enzymes and lipid peroxidation in hepatic mitochondria were measured.Mitochondrial membrane potential(MMP) and mRNA expression of CYP450 reductase in liver were determined by flow cytometer and in situ hybridization,and morphological changes of hepatocytes were observed under microscope with Oil-Red-O staining.Results ① In all rats treated with higher dose of VPA added with PB or not,there were no significant elevations of liver enzymes(ALT and AST).However significant abnormalities of function of blood coagulation and serum fibrinogen were shown, and the levels of plasma ammonia and L-carnitine were also changed significantly,and the changes were notable in infant rats or in those rats added with PB. ② Average contents of cytochrome aa3 in liver mitochondria of infant rats were reduced by 58.80% and 61.80% because of administration of high dose VPA and high dose VPA added with PB,but were reduced by 37.55% and 46.53% in adults.As for activities of SDH,which affected by high dose VPA in infants,were significant decreased by 44.8% and 57.9%,respectively,but still in normal range in adult groups.Activities of CCO in liver mitochondria were significantly lowered by high dose VPA or added with PB compared with controls(P
		                        		
		                        		
		                        		
		                        	
7.Analysis of peblA gene sequence and construction of eukaryotic expression recombinant plasmid of peblA gene of Campylobacter jejuni Pen:19
Hui ZHENG ; Fangcheng CAI ; Bing DENG ; Min ZHONG
Journal of Third Military Medical University 2003;0(07):-
		                        		
		                        			
		                        			Objective As the genome sequence of Campylobacter jejuni is a hypervariable sequence, the peblA gene sequence of Campylobacter jejuni Pen∶19 was sequenced and the peblA gene conservative was identified. To construct the eukaryotic expression recombinant plasmid of the peblA gene of Campylobacter jejuni Pen∶19 . Methods Total DNA of Campylobacter jejuni Pen∶2, Pen∶8, Pen∶19 and Pen∶21 as templates respectively, peblA gene DNA with KpnⅠ and EcoRⅠ sites was amplified by PCR using the same primer and the PCR product of Pen∶19 was sequenced. Because most close association with Guillain-Barre syndrome, the PCR product of Pen∶19 was selected as target gene and cloned into pcDNA3.1(+) and constructed the recombinant plasmid that was identified by endonuclease digestion and PCR and confirmed by sequencing. The recombinant plasmid was then transfected into mammalian cell COS-7 for expression. The transient expression was investigated by RT-PCR. The expression of peblA gene in the culture supernatant of positive clones was analyzed by ELISA. Results The peblA gene sequence of Campylobacter jejuni Pen∶2 and Pen∶19 was identical. The target gene was amplified from COS-7 cells transfected with recombinant plasmids by RT-PCR. PEBl protein could express in the culture supernatant in the transfected COS-7 cells by ELISA. Conclusion The peblA gene of Campylobacter jejuni was conservative. The recombinant plasmid was constructed and expressed in COS-7 cells successfully. The results obtained lay the foundation for research on development of Campylobacter jejuni DNA vaccine.
		                        		
		                        		
		                        		
		                        	
8.Comparsion of cerebral palsy teatments with Shangtian and Bobath method.
Banghui LI ; Yongping REN ; Fangcheng CAI
Chinese Journal of Tissue Engineering Research 2001;5(3):45-46
		                        		
		                        			
		                        			Objective The object of this study was to characterize the theraputic method of Shangtian and Bobath in treating cerebral palsy.Methods Sixty four patients with cerebral palsy,age from ,were subjected to this study.They were divided into two groups.treated with Shangtian method and Bobath method.Quantity evaluation on motion function was performed before and after the treatment.Results The overall score on motion function development was not significantly different between two groups(P >0.05).However,These were significantly different in seating function development and induces protective extensor thrust parachute reaction,between the two groups.Positive suppoting reaction improvement of 73% in shangtian treated group,62% in Bobath's treated group. Conclusion Both of the two therapeutic methods are effective in treating cerebral palsy.However.Shangtian is more effectual in improving seating function,postive suppoting reaction and induced protective extensor thrust parachute reaction.
		                        		
		                        		
		                        		
		                        	
9.Immunological mechanism of peripheral neuropathy induced by Campylobacter jejuni-Incidences of experimental peripheral neuropathy induced by Pen19 and Pen43
Chinese Journal of Pediatrics 2001;39(1):21-24
		                        		
		                        			
		                        			Recent studies have suggested that Campylobacter jejuni (CJ) may be the most common pathogen causing Guillain-Barre syndrome (GBS). Fifteen to seventy-five percent of patients with GBS had had a recent CJ infection. The serotypes of CJ isolated from stool usually are Penner (Pen) 19, Pen4, Pen41, and so on. Although some serotypes of CJ have been proved to associate with GBS, the directly evidence from the animal model is needed. Objective  To clarify the association of peripheral neuropathy with GBS by using the different serotypes of CJ to immunize Wistar rats respectively. Methods Sixty Wistar rats were divided into three groups: CJ-Pen 19 group, CJ-Pen43 group and normal saline control group. Each rat was immunized repeatedly with Freund′s adjuvant and inactivation antigen of CJ-Pen19, Pen43 and normal saline, respectively. The serum titers of specific anti-CJ antibodies were detected by ELISA. The pathologic examination of sciatic nerves was performed at different time points: 1, 2, 3, 4 and 5 weeks, respectively after the immunization. Results (1) Compared with the normal saline group, the serum titer of anti-CJ IgG elevated after immunization and sustained at a higher level after the 3rd-4th week in the CJ-Pen19 and Pen43 groups of rats. (2) The incidences of pathologic nerves and fibers in CJ-Pen19 group (60.0% and 17.7%, respectively) were much higher than those of CJ-Pen43 group (10.0% and 0.5%, respectively) and of normal saline group (5.0% and 0.5%, respectively, P<0.001). There was no significant difference between CJ-Pen43 and normal saline groups (P>0.05). (3) At the initial stage, the type of pathologic fibers was mostly axon degeneration. The ratio of axon degeneration to demyelination was 3.4% to 0.7%. But at advanced stage, the proportions of demyelination increased rapidly and the ratio of axon degeneration to demyelination was 5.3% to 24.1%. (4) There was a positive correlation between the antibody titer of specific anti-CJ IgG and the incidence of axon degeneration (r=0.801) compared with the demyelination. There was no correlation between the IgM level and the incidence of axon degeneration or demyelination (r=0.253 and 0.281, respectively). Conclusions  (1) The incidence of experimental peripheral neuropathy might depend on CJ serotypes; (2) The majority of pathologic fibers showed axonal degeneration at the initial stage and remarkable demyelination at the later stage. (3) The specific anti-CJ IgG antibody might act as an important pathogenic factor in the process of axon degeneration.
		                        		
		                        		
		                        		
		                        	
10.Hepatocyte growth factor modified human umbilical cord mesenchymal stem cell therapy promotes remyelination after intracerebral hemorrhage in rats
Anmin LIU ; Ming LUO ; Wangqing CAI ; Fangcheng LI ; Guo LI ; Waisang POON
Chinese Journal of Pathophysiology 2000;0(10):-
		                        		
		                        			
		                        			AIM:To study the effects of neurological improvement and remyelination after intracranial hemorrhage(ICH)in rats by a novel therapeutic strategy with hepatocyte growth factor(HGF)gene transfected human umbilical cord mesenchymal stem cells(hUCMSCs)by lentiviral vector.METHODS:ICH was induced in 60 adult male Sprague-Dawley rats by a stereotactically guided injection of bacterial type IV collagenase into the right internal capsule.Non-modified hUCMSCs,HGF-modified hUCMSCs with lentiviral vector or PBS were administered left intraventricularly 7 d after right internal capsule ICH.All rats underwent modified neurological severity scores for 35 d.Luxol fast blue staining,immunohistological staining and Western blotting assessments for myelin basic protein(MBP)were applied.RESULTS:The ICH rats receiving HGF-modified hUCMSCs demonstrated significant functional recovery,determined by modified neurological severity scores,compared to the other groups from 2 weeks after cell therapy.As indicated by Luxol fast blue staining,the percent area of demyelination was obviously reduced in the HGF-hUCMSC treatment group compared to the PBS control group and hUCMSC-only treatment group at 5 weeks after ICH.The expression of MBP detected by immunohistological staining and Western blotting was significantly higher in HGF-hUCMSCs treated brain than that in other groups(P
		                        		
		                        		
		                        		
		                        	
            
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