2.Effect of naloxone on neuronal cells apoptosis induced by repeated febrile seizures
ying, SHAN ; jiong, QIN ; xiu-ying, TANG ; xing-zhi, CHANG
Journal of Applied Clinical Pediatrics 2004;0(09):-
Objective To investigate the effect of naloxone on neuronal cells apoptosis induced by repeated febrile seizures(FS).Methods Warm water was used to induce 70 rats FS model 15 days after birth in this study; each rat was induced 7 times febrile seizures at one- day interval . Seventy rats were randomly divided into naloxone-treated group and FS control group, receiving injection of naloxone or saline at 5, 30, 60 min and 2 hours after FS each day respectively. The rats were sacrificed 24 hours after the last seizure. Neuronal cell apoptosis was determined by TUNEL methods in situ cell death kit. TUNEL positive cells(TPC) were stained and counted as apoptosis in hippocampus and cortex. Ultrastructural changes of apoptosis neurons were observed under the electron microscope(EM). Results Compared with the FS control group, naloxone treatment could significantly relieve neuron apoptosis induced by repeated FS when it was used at 5, 30, 60 min after the last FS. However there was no significant difference in neuron apoptosis between 2 groups when naloxone was used at 2 hours after FS. The comparison of different naloxone administration time showed that the earlier naloxone was injected,the fewer apoptosis neurons were induced by FS.Conclusion Naloxone,as early used in proper dosage,may significantly alleviate apoptosis after repeated FS ,and protect neurons.
3.Influence of endogenous nitric oxide synthase /nitric oxide system on brain damage induced by recurrent febrile seizures
zhi-xian, YANG ; jiong, QIN ; dingfang, BU ; junbao, DU ; zhi xing, CHANG ; ying, HAN
Journal of Applied Clinical Pediatrics 2004;0(12):-
Objective To explore the changing regularity of nitric oxide synthase(nNOS) in recurrent febrile seizures (FS), and the influence of NOS/NO on brain damage induced by recurrent FS.Methods FS rats were induced in a bath of warm water.The ex-periments were divided into 2 groups. The contents of nNOS cDNA in the first group was measured by quantitative RT-PCR and the contents of nNOS protein was measured by Western blot.The mtensity , latency, duration and rectal temperature of the seizure in rats in the second group were recorded. Morphologic changes of hippocampal neurons were observed with HE stain.Results Alter recur-rent FS, the expression of nNOS mRNA in hippocampus was significantly inereased compared with those in control group and hyper-thermia group, associated with an increase of nNOS protein.With the increase of seizure number,thert were changes of seizure latency and gradually prolonged trend of the seizure duration. By using the inhibitor of NOS, the seizure latency was gradually prolonged and the prolonged trend of the seizure duration was significantly decreased than that in FS group.There was no significantly difference of seizure intensity and rectal temperature between 2 groups.After recurrent FS, histological changes of hippocampal neurons could be seen under light microscope.The inhibitor alleviated nearonal injury.Conclusions Recurrent FS can induce nNOS gent expression.The NOS/NO system may be involved in the development of brain damage induced recurrent FS.
4.Nitric Oxide Regulated Expression of ?-Aminobutyric Acid B Recepto r Subunits during Recurrent Febrile Seizures
ying, HAN ; jiong, QIN ; ding-fang, BU ; zhi-xian, YANG ; xing-zhi, CHANG ; jun-bao, DU
Journal of Applied Clinical Pediatrics 2004;0(09):-
Objective To explore the effect of nitric oxide (NO) on ?-aminobutyric acid B receptor (GABA_BR) subunits during recurrent febrile seizures (FS).Methods Twenty-four Sprague-Dawley rats aged 21 days were randomly divided into 4 groups: control group (37.0 ℃ water,n=8), FS group (45.2 ℃ water,n=8), FS + SNP group (45.2 ℃ water,n=8), FS+L-NMMA group (45.2 ℃ water,n=8). FS rats were induced 10 times in a warm-water bath, once every 2 days. The plasma level of NO was detected by the spectrophotometer. The expressions of GABA_BR subunit mRNA and c-fos gene were examined by in situ hybridization. The expressions of GABA_BR subunit and Fos protein were observed by immunohistochemistry. Results The plasma level of NO increased in FS + SNP group while decreased in FS+L-NMMA group compared with that in FS group. The expressions of GABA_BR_2 were down-regulated in FS+SNP group, while GABA_BR_1 hardly changed compared with those in FS group. In FS+L-NMMA group, both the expression of GABA_BR_2 and GABA_BR_1 up regulated compared with those in FS group. The expressions of c-fos gene and Fos protein were significantly enhanced after recurrent FS. SNP elevated the expressions of c-fos gene and Fos protein, while L-NMMA down regulated the expressions of them.Conclusion NO may play a regulatory role through modulating GABA_BR function in the pathogenesis of recurrent FS.
5.Influence of Gamma-Aminobutyric Acid B Receptor on Expression of Carbon Monoxide/ Heme Oxygenase System during Recurrent Febrile Seizures
ying, HAN ; jiong, QIN ; ding-fang, BU ; zhi-xian, YANG ; xing-zhi, CHANG ; jun-bao, DU
Journal of Applied Clinical Pediatrics 2004;0(12):-
Objective To explore the influence of ?-aminobutyric acid B receptor(GABA_BR)on carbon monoxide (CO)/heme oxygenase(HO-1)system during recurrent febrile seizures (FS).Methods Sprague-Dawley rats aged 21 days were randomly divi- ded into 4 groups:control group and FS group,FS+baclofen group,FS+phaclofen group.FS in rats were induced 10 times in a bath of warm water, once every 2 days.The plasma level of CO was detected by the dual wave lengh spectrophotometer;the expressions of GABA_BR and HO-1 mRNA were examined by insitu hybridization;the expressions of GABA_BR and HO-1 protein were observed by immunohistochemistry.Results The plasma level of CO increased in FS+baclofen group,while decreased in FS+phaclofen group compared with FS group.The expressions of GABA_BR and HO-1 upregulated in FS+baclofen group,while decreased in FS+phaclofen group compared with FS group.There were significant difference (All P
6.Over-articular external fixator combined with limited internal fixation for the treatment of high-energy Pilon fractures.
Da-Cheng LIU ; Xiao-Ning YANG ; Chang-Zhi HUANG ; Yi-Gong SUN ; Xing-Ming DAI
China Journal of Orthopaedics and Traumatology 2014;27(4):331-334
OBJECTIVETo study clinical effects of the over-articular external fixator combined with limited internal fixation for the treatment of Pilon fractures caused by high energy.
METHODSFrom September 2003 to April 2011, 36 patients with Pilon fractures caused by high energy were treated with the over-articular external fixator combined with limited internal fixator. There were 25 males and 11 females, ranging in age from 16 to 72 years old,with an average of 38 years old. The diagnoses of all patients were determined by conventional X-ray examination or three-dimensional spiral CT examination. The AOFAS scoring criteria was used to evaluate the therapeutic effects. The patients with comminuted fractures were treated with screw or Kirschner wire fixation without uncovering periost so as to enhance stability between fracture end and bone blocks,followed by the fixation with over-articular external fixators.
RESULTSAll the patients were followed up, and the duration ranged from 4 to 27 months, with an average of 13 months. Thirty-two patients got wound healing at the first stage. And the bone union duration ranged from 2 to 6 months, with a mean of 3 months. According to the AOFAS ankle-hindfoot subjective scoring standard, 13 patients got an excellent result, 20 good and 3 fair, with an score of 88.2 +/- 3.6. Twelve patients had infections at pinhole, 5 patients had pinhole pain. One patient had the fixator broken induced by over loading, who was cured after treatment. There were no complications such as nerve or vascular injuries, or osteomyelitis.
CONCLUSIONThe over-articular external fixation combined with limited internal fixation for the treatment of Pilon fractures caused by high energy is an ideal method, which has such advantages as reliable fixation, simple operation, coincidence with principles of biomechanical fixation, and benefit for fracture healing.
Adolescent ; Adult ; Aged ; Ankle Injuries ; diagnostic imaging ; surgery ; Ankle Joint ; diagnostic imaging ; surgery ; External Fixators ; Female ; Fracture Fixation ; Fracture Fixation, Internal ; Humans ; Internal Fixators ; Male ; Middle Aged ; Radiography ; Treatment Outcome ; Young Adult
7.Therapy and Diagnosis of Phaeohyphomycosis of Central Nervons System
xing-zhi, CHANG ; jian - guo, LI ; ruo-yu, LI ; xin-hua, BAO ; zhe, WAN ; jiong, QIN
Journal of Applied Clinical Pediatrics 2003;0(10):-
Objective To explore the clinical characteristics ,diagnosis, treatment and prognosis of phaeohyphomycosis. Methods Clinical data were collected, including history, physical examination, cranial and spinal imaging. Brain biopsy was performed. Data of the pathology and incubation of brain tissue were analyzed. Responsiveness to treatment was followed up. Results A previously healthy three and half years old boy was presented to our unit, with a three- month history of recurrent headache, vomiting, progressive paraplegia accompanied by urinary continence and constipation. A computed tomogram scan and magnetic resonance imaging of the brain revealed multiple lesions located in the region of the parietal - occipital lobes, periventricular area and frontal lobe, with prominent surrounding edema and irregular peripheral enhancement of the mass after the administration of contrast materials. A cerebral biopsy was performed and the pathological report was cerebral phaeohyphomycosis. The culture of the tissue and cerebrospinal fluid grew a same fungus identified as exo-phiala dermatitidis. The patient's response to therapy was poor, the parents of the boy gave up therapy, and the boy died 1 month later. Conclusions Cerebral Phaeohyphomycosis caused by Exophiala dermatitidis is rare, but the most serious form of fungus infection. Pathology and incubation of the tissue are essential for diagnosis. There is no curative therapy and the prognosis is poor.
8.Initial Clinical and Imaging Characteristics of Basal Ganglia Germinoma in Children
xing-zhi, CHANG ; ye, WU ; xin-hua, BAO ; yue-hua, ZHANG ; jiong, QIN
Journal of Applied Clinical Pediatrics 2006;0(14):-
Objective To explore the initial clinical and imaging characteristics of basal ganglia germinoma in children.Methods Four patients with basal ganglia germinoma were reported.Their clinical features,laboratory findings,radiological manifestations,treatment and outcome were analyzed.They recieved radiation therapy and chemotherapy after diagnosis.All patients were clinically diagnosed,according to the results of low-dose cranial irradiation.The outcomes were followed up for 2 years.Results All patients were male and school-aged(9-13 years) children.The course of the disease ranged from 5 to 13 months.All patients were presented with slowly progressive hemiparesis,and 2 cases of them were presented with cognitive decline and psychosis.Seizure occurred in 2 patients.The serum ?-human chorionic gonadotropin(?-hcG) level was significantly increased in 2 patients(30.16 IU/L and 77.85 IU/L,respectively),and mildly elevated in 1 patient(4.29 IU/L),while serum ?-hcG level in another case was within normal control range.MRI demonstrated mildly high intensity in the left or right basal ganglia on T1-weighted and T2-weighted images without remarkable occupying lesion.Ipsilateral hemiatrophy of the hemisphere and midbrain was also noted.Inhomogeneous Gd-DTPA enhancement was observed.All patients had been treated with radiation therapy and chemotherapy.During 2 years follow up,significant improvement was observed in all patients after therapy,imaging lesions disappeared and the elevated ?-hcG level of those elevated before therapy returned to normal.Conclusions Early diagnosis and treatment for basal ganglia germinoma are critically important to improve the prognosis.In young male patients with progressive hamiparesis,basal ganglia germinoma should be considered for differentiation,if abnormal high intensity signals in basal ganglia on T1-weighted and T2-weighted image with ipsilateral hemiatrophy of the hemisphere are demonstrated on MRI,even without occupying effect.
9.Neuroprotective effect of naloxone in brain damage caused by repeated febrile seizure.
Ying SHAN ; Jiong QIN ; Xing-zhi CHANG ; Zhi-xian YANG
Chinese Journal of Pediatrics 2004;42(4):260-263
OBJECTIVEThe brain damage caused by repeated febrile seizure (FS) during developing age is harmful to the intellectual development of children. So how to decrease the related damage is a very important issue. The main purpose of the present study was to find out whether the non-specific opiate antagonist naloxone at low dose has the neuroprotective effect on seizure-induced brain damage.
METHODSWarm water induced rat FS model was developed in this study. Forty-seven rats were randomly divided into two groups: normal control group (n = 10) and hyperthermic seizure groups (n = 37). The latter was further divided into FS control group (n = 13) and naloxone-treated group (n = 24). The dose of naloxone is different in two naloxone-treated groups (12/each group), in one group the dose was 1 mg/kg, in the other one 2 mg/kg. Seven febrile seizures were induced in each rat of hyperthermic seizure groups with the interval of 2 days. The rats were weighed and injected intraperitoneally with naloxone once the FS occurred in naloxone-treated group, while the rats of the other groups were injected with 0.9% sodium chloride. Latency, duration and grade of FS in different groups were observed and compared. HE-staining and the electron microscopy (EM) were used to detect the morphologic and ultrastructural changes of hippocampal neurons.
RESULTSIn naloxone-treated group, the rats' FS duration and FS grade (5.02 +/- 0.63, 2.63 +/- 0.72) were significantly lower (t = 5.508, P < 0.01; t = 8.439, P < 0.01) than those in FS control group (7.70 +/- 2.25 min, 4.52 +/- 0.49), although no significant gap was observed on FS latency between them. In FS control group, HE-staining pattern of hippocampal CA(1) and CA(2) showed lots of disordered neurons with confused polarity and vacuoles formed. Nuclei were with various size, some rounded and some oblong. While in naloxone-treated groups, the arrangement of neurons was regular, only a small quantity of neurons had changed polarity and vacuoles formed. Most nuclei were oblong and in the same size. In hippocampal CA(1) region and dentate gyrus of rats from FS control group, EM showed that the most mitochondrion volumes obviously increased with vacuoles formed, the matrix condensed, the ridge obscured or disappeared, apoptosis body emerged. Minor to moderate dilation of rough endoplasmic reticulum and Golgi's complex was also observed. However, in naloxone-treated groups, the number of neurons with swollen mitochondrion and endoplasmic reticulum was much fewer than that in FS control group. No apoptosis body was observed. The comparison between them showed much lighter brain damage in naloxone-treated groups than in FS control group.
CONCLUSIONAlthough low-dose naloxone could not totally stop the occurrence of febrile seizure, it could lighten the brain damage resulted from repeated FS to some extent.
Animals ; Brain ; drug effects ; pathology ; Male ; Models, Animal ; Naloxone ; pharmacology ; therapeutic use ; Narcotic Antagonists ; pharmacology ; therapeutic use ; Neuroprotective Agents ; pharmacology ; therapeutic use ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Seizures, Febrile ; drug therapy ; physiopathology ; Treatment Outcome
10.Repair of damaged intestinal mucosa in a mouse model of sepsis
Rui-Ming CHANG ; Li-Qiang WEN ; Jian-Xing CHANG ; Yu-Ru FU ; Zhi-Peng JIANG ; Shuang CHEN
World Journal of Emergency Medicine 2013;4(3):223-228
BACKGROUND:The intestine is not only the main target attacked by sepsis but also the vital organ which mediated sepsis. The recovery of the damaged intestinal barrier structure and function is related to the occurrence and outcome of multiple organ dysfunction syndrome (MODS). How to protect and reduce the damage of the intestinal mucosa and how to promote the reconstruction of the intestinal mucosa have been the important topics in sepsis for many years. This study aimed to investigate the influential factors of intestinal mucosal reconstruction after intestinal epithelial injuryin vivo in a mouse model of sepsis.METHODS:Mice were subjected to cecal ligation and puncture (CLP) for induction of sepsis to assess intestinal mucosal damage, epithelial cell apoptosis, and transformed number of goblet cells, and to detect the concentration of TNF-α, IL-1 and TGF-β1 and TFF3 (trefoil factor 3) expression in the small intestinal mucosa. All above were performed by HE staining, western blot, ELISA and immunohistochemistry respectively. The experimental animals were divided into a sepsis group and a sham-operation group. The animals with sepsis were separately killed at 6 (7 animals), 24 (7 animals) and 48 hours (7 animals) after CLP.RESULTS:Injured intestinal mucosa was observed in the 3 groups under a light microscope, in which damage scores in the 24-hour and 48-hour groups were higher than in the 6-hour group and no difference was found between the two groups. Moreover, less of goblet cells or other epithelial cells adjacent to the injured surface migrated into the wound to cover the denuded area. The number of goblet cells was substantially decreased in the three CLP groups compared with the sham-operation group. Protein levels of IL-1 and TNF-α were significantly increased by 3-4 fold at all time points when compared with the sham-operation group, and cleaved caspase-3 by 4 fold. Although TFF3 expression was modestly increased for 6 hours after the onset of CLP, it appeared to decline at 24 hours and 48 hours as shown by Western blot. A similar tendency was observed upon TGF-β1, i.e. the protein level was not elevated at 24 hours and 48 hours, but increased modestly at 6 hours.CONCLUSIONS:Sepsis from CLP shows less restitution on the surface of injured intestinal mucosa. There is evidence that both constant inflammatory reaction and epithelial cell apoptosis may affect mucosal reestablishment of the intestine at the onset of sepsis. Mucosa after severe sepsis showed the state of high inflammation, and declined goblet cell function and mucosal reconstruction, which affected the repair of damaged intestinal barrier. Constant inflammatory reaction, and declined goblet cell function and mucosal reconstruction ability may affect the reestablishment of intestinal mucosa at the onset of sepsis.