Effects of immunoglobulin on the neuronal expression of IL-1beta and IL-1ra and the neuronal death at hippocampus in rats with convulsion induced by pentylenetetrazol / 中华儿科杂志

<p><b>OBJECTIVE</b>To study the effects of immunoglobulin on the neuronal expression of IL-1beta and IL-1ra and the neuronal death at hippocampus in rats with convulsion induced by pentylenetetrazol.</p><p><b>METHODS</b>The epilepsy model was established by injecting intraperitoneally pentylenetetrazol (PTZ) into Wistar rats. Forty-five rats were randomly divided into three groups, normal control group, PTZ plus intravenous immunoglobulin (PTZ-IVIG); PTZ plus normal saline (PTZ-NS). Neuronal death was assessed by light microscopy with the hematoxylin-eosin (HE) staining and with in situ terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL). IL-1beta and IL-1ra expressions were examined by histochemistry.</p><p><b>RESULTS</b>The ratio of IL-1beta/IL-1ra at hippocampal CA(1) region in PTZ-IVIG group (0.5 +/- 0.1) was significantly lower than that in PTZ-NS group (1.9 +/- 0.5, t = 12.9, P < 0.05). Apoptotic cell numbers at the hippocampal CA(1) region were significantly decreased in the PTZ-IVIG group, compared to PTZ-NS group (t = 27.1, P < 0.05). The numbers of positive cells were 16.4 +/- 3.3/1000 microm(2) in the former and 41.7 +/- 3.5/1000 microm(2) in the latter. Necrotic cell numbers at the hippocampal CA(1) region were significantly decreased in the PTZ-IVIG group (19.0 +/- 2.6/1000 microm(2)), compared to PTZ-NS group (42.3 +/- 4.9/1000 microm(2), t = 20.9, P < 0.05).</p><p><b>CONCLUSION</b>Immunoglobulin could inhibit neuronal death induced by convulsion and its possible mechanism might be the regulation of IL-1 system in neurons.</p>

Relationship between programmed cell death mechanisms and neuronal necrosis induced by seizures / 中华儿科杂志

<p><b>OBJECTIVE</b>To clarify if programmed cell death mechanisms induced by seizures take part in the necrotic process of neurons.</p><p><b>METHODS</b>Seizure was induced by pilocarpine (P) in Sprague-Dawley adult rats which were allowed to recover for 24 or 72 hours before perfusion-fixation. Neuronal death was assessed by light microscopy with the hematoxylin-eosin (HE) staining and with in situ terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL). Bax and Bcl-2 protein expression were examined by histochemistry.</p><p><b>RESULTS</b>Twenty-four and 72 hours after seizures, neuronal death in hippocampus CA1 region was morphologically necrotic. TUNEL-positive and morphologically necrotic cells increased in the hippocampal CA1 region at 72 hours after seizures, there was significant difference compared with controls (P < 0.001). Bax expression was also increased in the hippocampal CA1 region at 72 hours after seizures (P < 0.001), but Bcl-2 expression did not increase, while Bcl-2/Bax ratio decreased.</p><p><b>CONCLUSION</b>Seizures induced late-onset neuronal necrosis was accompanied by programmed cell death mechanisms.</p>

The Neuroprotective Effect of Growth Hormone on Neuronal Injury of Brain in Pilocarpine induced Status Epilepticus / 대한내분비학회지

BACKGROUND: Several growth factors, including growth hormone (GH) and Insulin like growth factor-I, have been reported to have a neuroprotective effect in experimental models of hypoxic ischemia. This study is aimed at assessing the clinical significance of growth hormone for neuroprotection in status epilepticus induced neuronal cell deaths. METHODS: Pilocarpine induced status epilepticus (SE) was studied in rats (male, Sprague-Dawley). Rats were divided into pre- or post-treatment groups that had either a low (5 U/kg/day) or high (10 U/kg/day) dose of recombinant human GH (Eutropin, LGCI, Korea), and then subdivided into 24 hour, 72 hour and 1 week groups. This was done in the pretreatment groups for 5 days before SE and in the post-treatment groups for 5 days after 2 hrs of SE injection, after SE, the GH was daily injected via intraperitoneal route. Status epilepticus was induced by pilocarpine (360 mg/kg) with scoplamine (1 mg/kg) 30 minutes before pilocarpine injection using a stereotaxic instrument and EEG monitoring. Rats were killed at 24 and 72 hours after the SE in the pretreatment groups and at 1 week after the SE in the post-treatment groups for pathology studies. Neuronal injuries in the rat brain were studied by Hematoxylin & Eosin stain and the TUNEL method. RESULTS: Neuronal necrosis was found in the hippocampal CA1 and CA3 regions in all experimenatal groups after SE, and was more severe in the CA3 region. Apoptosis was found only in the pre-GH treated group and there were TUNEL-positive and morphologically necrotic cells in the hippocampal CA1 and CA3 regions at 72 hours after SE. Neuronal necrosis and apoptosis were significantly decreased in the high dose GH treated groups (p0.05). CONCLUSION: Growth hormone has a neuroprotective effect in neuronal cell death (necrosis and apoptosis) that is caused by pilocarpine induced status epilepticus in a dose dependent manner and prevents the activation of apoptosis by SE in neurons which eventually become necrotic.

The Effect of MK 801 on the Development of Brain Damage, Spontaneous Recurrent Seizures and Mossy Fiber Sprouting in the Pilocarpine Induced Status Epilepticus Animal Model

PURPOSE: We investigated the effect on MK 801 on the development on brain damage, spontaneous recurrent seizures and mossy fiber sprouting in the pilocarpine induced status epilepticus animal model. Methods: Fifty two adult Sprague Dawley male rats(180-240gm) were studied under ketamine/xylazine(87mg/13mg/kg, IP) anesthesia and were implanted at the F3, P3, F4, P4 areas for recording EEG. With a single intraperitoneal(IP) administration of pilocarpine hydrochloride(360mg/kg), 70% developed status epilepticus(SE). When SE was not induced within 1 hour after injection of pilocarpine, the second dose of pilocarpine(175mg/kg, IP) was injected, with 86.6% of success. Results: All studied animals were divided into two large groups, one group was treated with NMDA receptor antagonist, the other was control group. The mean duration of SE was 62.00+/-6.80 minutes in the MK 801(1mg/kg, IP, 30 minutes after SE) treated group, and 61.10+/-7.37 minutes in the control group without any signigicant differences(P>0.05). Neuronal loss(necrosis dominantly) was observed at CA1 and CA3 areas in the control group, with more loss after 6 weeks than 24 or 72 hrs specimens. However, there was no neuronal loss in MK 801 treated group. The protective effect of MK 801 for neuronal injury suggested the glutamate receptor activation was involved in the neuronal injury induced by repeated seizure attack. Spontaneous recurrent seizures(SRS) were observed 70% of animals in the control group, but there were no SRS observed in the MK 801 treated group. The mean scores of mossy fiber sprouting were significantly higher in the control group(2.05+/-0.47) than MK 801 treated group(0.4+/-0.32)(P<0.05). Conclusion: These results suggested that SRS and mossy fiber sprouting were associated with NMDA receptor activation, and NMDA receptor activation had a key role in the epilepsy development.

Role of Sp Family Transcription Factors in Growth Hormone Receptor Gene Expression / 대한소아내분비학회지

PURPOSE: The growth hormone receptor(GHR) is essential for the actions of growth hormone on postnatal growth and metabolism. GHR transcripts are characterized by the presence of disparate 5'untranslated exons. In contrast to L1 transcript, factors regulating the expression of the GC rich L2 transcript have remained unidentified. The purpose of this study is in order to characterize the mechanisms regulating expression of the L2 transcript in the murine GHR gene METHODS: Transient transfection experiments including deletional analysis and co-transfection assay were performed to find a region containing promoter activity in the L2 5'flanking sequence using BNCL2(mouse liver) cells, CV-1(African green monkey kidney) cells, HRP.1 trophoblasts and Drosophila Schneider(SL2) cells. Sequencing analysis was performed to find the region contained consensus binding sites for transcription factors. Standard gel shift(Electrophoretic mobility shift assay, EMSA) and supershift analysis using liver nuclear extracts was performed to establish proteins(transcription factors) bound this regulatory element. RESULTS: The 5'flanking region of the L2 untranslated region(UTR) exhibited promoter activity in BNCL2(mouse liver), CV-1(monkey kidney) cells and HRP.1 trophoblasts. Deletional analyses indicated the presence of a Sp binding site important for transcription of the L2 UTR and localized the major regulatory region within 75 bp of the 5'transcription start site. Sequencing analyses revealed the region contained consensus binding sites for the Sp family of transcription factors. EMSA and supershift EMSA revealed that in mouse liver nuclear extracts that Spl and Sp3 bound to this cis-element. Functional studies in Drosophila SL2 cells and BNCL2(mouse liver) cells established the ability of Sp3 and Sp1 to stimulate transcriptional activity via this cis-element. Functional studies in Drosophila SL2 cells demonstrated a functional interaction between Sp3 and Sp1 at this DNA-binding site. CONCLUSION: Sp family transcription factors play a role in regulation of L2 transcript gene expression in the 5'flanking region of the murine GHR gene.

Generalized Epilepsy with Febrile Seizures Plus(GEFS+) and a Mutation in the Voltage-gated Sodium(Na+)-Channel Beta 1 Subunit Gene(SCN1B)

PURPOSE: It was reported that gene locus for generalized epilepsy with febrile seizures plus(GEFS+) exist in chromosome 19q13.1, and has relationship with a 387 C G mutation in the SCN1B gene. This study is to determine whether there is the 387 C G mutation in the children with GEFS+ and simple febrile seizures(FS). METHODS: The sample group consisted of 16 patients with GEFS+ and 10 patients with FS who were diagnosed by our department of pediatrics from Jan. 1998 to Dec. 1999. The control group consisted of 15 children who do not have seizure disorders. Genomic DNA was extracted from peripheral blood and a segment of the SCN1B exon 3 was amplified by PCR technique. Purified PCR products were treated with restriction enzyme, Hin P1. The restriction pattern was analyzed by sequencing analysis. RESULTS: Sixty nine%(11 of 16) patients with GEFS+ had family history for epilepsy, and epilepsy phonotypes were generalized tonic-clonic seizures in 82%(13 of 16), on the other hand 12%(2 of 16) and 6%(1 of 16) had absences and atonic seizures respectively. EEG findings showed generalized spike and wave in the all patients with GEFS+. in this study, however we could not observe a 387 C-->G mitation of the SCN1B in the children with GEFS+ and febrile seizures. CONCLUSION: The gene for GEFS+ may have a heterogenetic characteristics, and there may be racial differences in mutation frequency. Expanded studies involving large number of different families are required.

A Case of Tuberous Sclerosis Associated with Multiple Tumorous Condition

Tuberous Sclerosis is a neurocutaneous syndrome, which is characterized by seizure, mental retardation, angiofibroma and various tumorous conditions. We report a case of 15 year old girl with Tuberous Sclerosis associated with multiple tumorous conditions. We report this case with brief review of related literatures.

Analysis of Brain Evoked Potential Study in Cerebral Palsy Patients

PURPOSE: Cerebral palsy is a group of conditions characterized by nonprogressive motor and posture dysfunction developing during perinatal period due to brain damage. Combined sensory and cognitive disorders can evolve the secondary mental retardation or speech disorder. Brain evoked potential can evaluate the visual, auditory, somatosensory neuropathway, and the response of frontal, temporal, occipital lobe. We studied the usefulness of brain pvoked votential as a tool in the early diagnosis and treatment of sensory disorders in cerebral palsy. METHODS: We retrospectively reviewed the records of 86 cerebral palsy patients who were practiced brain evoked potential study in Chungnam National University Hospital from July, 1995 to June, 1999. We analyzed the visual, auditory, somatosensory evoked potential result and the correlations between the electroencephalography, radiologic brain imaging study and the brain evoked potential. RESULTS: 1) Clinical types of cerebral palsy were spastic type(85.0%), athetoid type(3.5%), mixed type(3.5%) and the remaining cases did not manifest any one the types above. 2) Abnormal evoked potential fingings were 25 cases(29.4%) in visual evoked potential, 16 cases(18.8%) in auditory evoked potential, 28 cases(37.8%) in median nerve evoked potential, 39 cases(52.7%) in tibial nerve evoked potential. 3) Electroencephalography, radiologic brain imaging study manifested no statistically significant correlations with the brain evoked potential result(P>0.05). CONCLUSION: As a noninnvasive neurophysiologic study, Brain evoked potential is a useful method predicting neurologic developmental progress and helpful to early diagnosis of sensory disorder in cerebral palsy patients.