Objective:To investigate the expression of estrogen receptor(ER) and interleukin-6(IL-6) in the intra-cardiac ganglia of rats and to identify the coexistence of these two proteins in the intra-cardiac ganglia.Methods:The double labeling techniques of immunocytochemical staining were conducted in the study.Results:There were three immunoreactively-labeled neurons in the ganglia.The ER-staining was showed brown in color,locating in the nucleus;IL-6 staining was showed red in color,locating in the cytoplasm;there were ER/ IL-6 double-labeled cells in same section,which accounted for about 20% to 30 %of the total single and double labeled neurons.Conclusion:ER and IL-6 can coexist in same rat intra-cardiac ganglia cells.Therefore,it provides morphological evidence for the theory about immune-neuro-endocrine network at the cellular level.

Objective:To study the expression and variance of interleukin-6(IL-6)and nerve growth factor positive neurons in the intrinsic cardiac ganglia of females that were intact,females that were ovariectomized,or ovariectomized females that were treated with 17?-estradiol.Methods:Immunohistochemical staining technique.Results:IL-6-IR and NGF-IR neurons were presented in the intrinsic cardiac ganglia of these groups,but the number of the IL-6-IR neurons increased remarkably and their expression increased obviously in the intracardiac ganglia of ovariectomized rats,while the number of the NGF-IR neurons decreased remarkably and their expression reduced obviously in the intracardiac ganglia of ovariectomized rats.Conclusion:IL-6 and NGF are present in rat intrinsic cardiac ganglionic cells and they are also influenced by estrogen,suggesting that 17?-estradiol may have the capacity to regulate the expression of IL-6 and NGF in intrinsic cardiac ganglia.

Objective To explore the correlation between serum myocardial enzymes and children with viral hepatitis according to clinical observation and analysis,in order to guide clinical treatment.Methods 66 children with viral encephalitis admitted in Women and Children's Hospital of Ninbo City from September 2010 to December 2011 were used as observation group,the aspartate a minotransferase(AST)level,lactate dehydrogenase (LDH)level,creatine kinase levels(CK)level,MB isoenzyme of creatine kinase(CK-MB)level in serum were detected by Olym pus AU-1000 automatic biochemical analyzer.The level ofα-hydroxybutyric acid dehydrogenase(α-HBDH)was detected,too.At the same time,another 48 normal children in our hospital were used as control group,and the same indexes were detected and recorded.After obtain those datas in two groups,the differences of myocardial enzymes between observation group and control group,acute and convalescent,light and heavy encephalitis encephalitis, gammaglobulin gammaglobulin treatment and non-treatment group were compared and analyzed.Results showed that the degrees of myocardial damage were varied in children with viral inordinately,the more serious illness in children,the higher myocardial enzyme levels.Correspondingly,while the children with encephalitis were in recovery ,their cardiac enzyme levels were found significantly reduced.Adding gamma globulin may help cardiac enzymes decreasing at a faster rate,favoring patients' recovery.Conclusion Cardiac enzyme activity can be used as an important indicator of clinical observation in children with viral and their prognosis,proper treatment should be carried out when the relationship between serum myocardial enzymes and viral encephalitis are clear.

Objective To observe the expression of TLR4 and NF-κB in hippocampus injury of status epileptic rats and to study the regulating effect of PDTC on TLR4/NF-KB signal pathway and hippocampus injury, and to explore the role of TLR4/NF-κB signal pathway in the hippocampus injury of SE rats. Methods A hundred and six male Sprague-Dawley (SD) rats were randomly divided into control group (A), convulsion group (B), PDTC group (C), and group B were randomly divided into 4 subset groups (B1-B4), which would be executed at 4, 24, 48 and 72 hours after convulsion. Continuous epilepticus was induced by injecting lithium chloride and pilocarpine, and group C were daily injected with 100 mg/kg PDTC 30 minutes after convulsion stopped for 3 days. Then the histopathology changes in hippocampus were viewed by HE staining, TLR4 and NF-κB/p65 protein were detected by immunohistochemistry (IHC), the expression of TLR4 mRNA were detected by RT-PCR. Results Neuronal injury was observed after a long time of convulsion, and the change was increased gradually 72 hours after seizure, which was milder in group Cthan in group B4. The expression of TLB4 protein in group B (B1-B4 were 0.1287±0. 0260, 0. 1296± 0. 0285, 0. 1330±0. 0329 and 0. 1604±0. 0457, respectively) was significantly higher than in group A (0.0964±0.0324, t =0.0641-0.3236, all P<0.05), and that in group C (0.1271±0.0330) was much lower than in B4 group (t = -0. 0334, P <0. 01). The IHC staining of NF-κB/p65 showed that hippocampal neurons had positive expression in cell nucleus in group B compared with the group A (P < 0. 05), and the expression of NF-κB/p65 protein in group C was much lower than that in group B4 (P < 0. 01). The mRNA expression of TLB4 in rat hippocampus of group B were significantly elevated than that in group A (0. 268±0. 072, P < 0. 05), and the tendency was increased gradually, reaching the peak at 72 hours after seizure (1. 242±0. 100), and that in group C (0. 984±0. 263) was much lower than that in B4 group (t=-0.2578, P<0.01). There was a coincidence between the expression of TLB4 and NF-κB/ p65. Conclusions The increased expression of TLR4 and NF-κB/p65 in SE rat hippocampus may play an promotion role on the development of the hippocampus injury; PDTC can down regulate the expression of TLR4, and lessen the pathologic changes of hippocampus.

Objective To study the change of BDNF/NF-κB signal pathway induced by repeated febrile seizures.Methods Fifty-one 21 d male rats were randomly divided into three groups:normal control group(n=14),hyperthermal control group(n=19)and febrile seizure group(n=18).The febrile seizure model was devdoped by warrn water immersion[(45.0±0.3)℃].BDNF and NF-κB were measured by enzyme hnked immunosorbent assay(ELISA)and immunohistochemistry.Results The levels of BDNF in serum and hippocarnpus in FS group were higher than those in NC group(P＜0.01)and in HC group(P＜0.01).In FS group,the OD of the BDNF positive neuron was higher than that of NC group(P＜0.01)and HC group(P＜0.01);the OD of the NF-κb positive neuron in FS group was higher than that of NC group (P＜0.01)and HC group(P＜0.01),the OD of the NF-κB positive neuron in HC group higher more than in NC group(P＜0.01).There was positive correlation between BDNF expression and NF-κB activation(r=0.78.P＜0.01).Conclusion The expression of BDNF and NF-κB were obviously increased in FS group and they are positively correlated,suggesting that the BDNF/NF-κB signal pathway may be activated after febrile seizure.

[ ABSTRACT] AIM:To investigate the effect of edaravone ( ED) on the expression of caspase-3 and caspase-12 in the juvenile rat hippocampus after status convulsion ( SC) .METHODS: Juvenile male Sprague-Dawley rats were ran-domly divided into normal saline ( NS) control group, SC group and ED treatment group.The rats in each group were fur-ther divided into 5 subgroups according to different time points.The rats in SC group were kindled into epilepsy by lithium-pilocarpine chemical method.The protein expression of caspase-3 and caspase-12 was determined by immunohistochemistry methods.The mRNA expression of caspase-3 and caspase-12 was detected by RT-PCR.RESULTS:(1) The IA value of caspase-3 positive cells in 24~72 h SC group increased compared with NS group.With ED intervention, the IA value of caspase-3 positive cells decreased as compared with 48~72 h SC group.The results of RT-PCR showed that the mRNA ex-pression of caspase-3 was similar to the changes of protein.( 2 ) The results of immunohistochemistry showed that the IA value of caspase-12 positive cells in 12~72 h SC group increased compared with NS group.With ED intervention, the IA value of caspase-12 positive cells decreased as compared with 24~72 h SC group.The results of RT-PCR showed that the mRNA expression of caspase-12 was similar to the changes of protein.( 3 ) In ED group, Ⅴ grade convulsion was lower than that in SC group, and the latent period of seizures in ED group was significantly longer than that in SC group.CON-CLUSION:Edaravone inhibits the expression of caspase-3 and caspase-12 in pilocarpine-induced seizures in rat hippo-campus, suggesting that edaravone has protective effect against the damage caused by status convulsion.

Propedium iodide (PI) and bisbenzimide (Bb) were injected into anterior wall of the cardiac ventricle and lesser curvature of the stomach in the rat respectively. We have found that PI labelled cells are distributed in C_4-T_(12), Bb labelled cells in T_4-L_1, PI-Bb double labelled cells in T_6-T_(11) dorsal root ganglia. This result indicate that dichotomizing branches of peripheral process from some of dorsal root ganglionic neuron projecte to two visceral organs simultaneously. By means of immunocytochemistry (PAP method), some of double fluorescence cells are CGRP positive. The significance of the viscero-visceral sensory converging to same primary sensory neuron was discussed.

The primary sensory neuron simultaneously innervating two or three viscera was studied in this paper, In one group the viscera in neck, thorax, abdomen and pelvis of the rat were paired off, i.e. esophagus-thyroid gland; esophagus-trachea; heartstomach; lung-stomach; left kidney-spleen; urinary bladder-uterus; urinary bladderrectum; uterus-rectum. Propedium iodide (PI) or fast blue (FB) and bisbenzimide (Bb) or nuclear yellow (NY)were injected separately into the first and second organ of paired viscera.PI-Bb (or FB-NY) double labelled cells were observed in corresponding ganglia (dorsal root ganglia and upper and lower ganglia of vagus nerve). In another group, PI, FB and NY were injected into urinary bladder, rectum and uterus separately. We observed a very few of PI-FB-NY triple labelled cells in L_(1-2) dorsal root ganglia.The above findings indicated that there are sensory convergences of visceroviscera in the same sensory ganglion cell in rat.

Objective To demonstrate simultaneously on transcription and translate levels the tyrosine-hydroxylase(and somatostatin)gene expression and peptide storage in same intrinsic cardiac ganglionic neuron of bufo bufo gargarizans. Methods Using immunohistochemical method,in situ hybrodization method and in situ hybrodization/immunohistochemical double labeling method on freeze-sections of sinus venanum cavanum of bufo bufo gargarizans atrial posterior wall (30*!?m)to observe THmRNA and /or TH-IR and SSmRNA and /or SS-IR dyeing neurons of intrinsic cardiac ganglia. Results There are kinds of positive neurons:1.TH-IR positive neurons;2.THmRNA positive neurons;3.TH mRNA/TH-IR double positive neurons and 1.SS-IR positive neurons;2.SSmRNA positive neurons;3.SSmRNA/SS-IR double positive neurons.Conclusion 1.TH mRNA and TH appearing in same intrinsic cardiac ganglionic neuron demonstrate that a part of intrinsic cardiac ganglionic neurons are possibly adrenergic sympathetic neurons;2.SSmRNA positive neurons and SSmRNA/SS-IR double positive neurons indicat that SS could be a neurotransmitter of intrincic cardiac ganglinic neurons.It provides neuroanatomy evidence of regulating cardiac function(excitatory conduct、cardiac muscle secrete and contract)by intrinsic cardiac SS-nergic neuron.

[ABSTRACT]AIM:Tostudytheeffectsofbaicalin(BC)onglialfibrillaryacidicprotein(GFAP)andnuclear factor-κB ( NF-κB) expression and neuronal apoptosis in juvenile rat hippocampus after status convulsion ( SC) .METH-ODS:One hundred and ninety five juvenile male Sprague-Dawley rats were randomly divided into 3 groups:normal saline pretreatment group (NS group), SC group and SC with BC pretreatment group (BC group).Each of these 3 groups would be subdivided into 5 subgroups sacrificed at 4 h, 12 h, 24 h, 48 h and 72 h after SC.The rat SC model was prepared by lithium-pilocarpine chemical method .The protein expression of GFAP and NF-κB was detected by the method of immuno-histochemistry .The mRNA expression of GFAP was detected by RT-PCR.The neuronal apoptosis was observed by TdT-mediated dUTP nick end labeling ( TUNEL ) .RESULTS: Compared with NS group , the GFAP positive cells was in-creased in SC group (P<0.05).Compared with SC group, the expression of GFAP was significantly reduced in BC group (P<0.05).Compared with NS group, the NF-κB positive cells was increased in SC group (P<0.05).Compared with SC group, the expression of NF-κB was significantly reduced in BC group .RT-PCR showed that the expression trend of GFAP mRNA was similar to that of the protein .Compared with NS group , the TUNEL positive cells in the hippocampus CA1 area in SC group increased significantly 12 h after SC (P<0.01), and reached a peak at 48 h.After the intervention with BC, the TUNEL positive cells decreased significantly between 12~48 h after SC (P<0.05 or P<0.01), but the number of TUNEL positive cells remained significantly greater than that in NS group (P<0.05).CONCLUSION: The expression of GFAP and NF-κB in the hippocampus increased after SC in rats .Baicalin decreases the expression of GFAP and NF-κB in hippocampus of rats with pilocarpine-induced seizures , and reduces the number of neuronal apoptosis , sug-gesting that baicalin may protect against the brain damage caused by status convulsion .