The chemical nature of spinal ganglionic neurons, the peripheral processes of which project divergently to the somatic and visceral areas, has been identified by means of tri-labeling method of combining fluorescein tracing and immunocytochemistry. Ten rats were used. First, 2?l of 2% fast blue (FB) were injected into the left coeliac ganglion. Two days later, 2% nuclear yellow (NY) was injected into left 9-11th intercostal nerves(l?l for each). On the 4th day, animal was perfused with 10% formalin in 0.1mol/L phosphate buffer.The left Th9-11 spinal ganglia were removed and cut into sections by cryostat. The sections were observed under fluorescence microscope and photographed. The results showed that there were three kinds of neurons in the spinal ganglia: (1) single FB labeled cells with blue fluorescent cytoplasm accounted for 38.8% of total labeled cells; (2) single NY labeled cells with yellow fluorescent nuclei accounted for 52,7%; (3) FB and NY double labeled cells accounted for 8.5% and mostly were small or medium in size. Then, the double labeled cells-containing sections were further processed by substance P-demonstrating PAP immunocytochemical staining. The immunostain and fluorescent photographs in the same section were compared and identified each other. We have found that the labeling ratio of SP/NY was 1.4%; SP/FB was 7% and SP/NY+FB was 28.8%. Present study has not only identified the convergence of somato-vesceral sensation in spinal ganglia but also detected the chemical nature of these neurons(substance P) for the first time. In addition, this result has provided a morphological basis for the mechanisma of referred pain and somato-visceral reflection.

The divergent projections in the bulbo-cerebellar system of rats were studied by double fluorescein labeling method. 2 ?1 of 2％ Fast blue (FB)were injected into the cerebellar cortex of left anterior lobe, 32 hours later, 2 ?l of 2％ Nuclear yellow (NY)were injected into the cerebellar cortex of right anteior lobe. The animals were allowed to survive 16 hours thereafter. Intracardiac perfusion were performed with 10％ formalin in 0.1 mol/L phosphate buffer. The cerebellum and medulla oblongata were removed and serial sections at 20 ?m were cut by cryostat. One of every two sections was preserved for fluorescence microscopic observation. The results identified that there was no intereonnection between the anterior lobes of two cerebellar hemispheres and the anterior lobe of cerebellum received afferent projections from much more nuclei of the medulla oblongata than previous description. The FB or NY single labeled and FB/NY double labeled cells were observed in the nucleus reticularis lateralis, subnucleus reticularis ventralis, nucleus raphe obscurus, nucleus prepositus hypoglossi, nucleus cuneatus lateralis, nucleus spinalis nervi trigemini and nucleus olivaris aceessorius dorsalis. The double labeled cells accounted for 23％ of total labeled cells except the nucleus olivaris inferior. The results mentioned above suggested that the cerebellum receive multiple afferent connections originated from the medulla oblongata, and the divergent projections further amplify the information-receiving areas in the cerebellum, and synchronize the activities of both cerebellar hemispheres.

In order to clarify the regulatory mechanism of the neurohormone releasing in the neurohypophysis, the immunohistochemical and chemical lesion method were combined to demonstrate the vasopressin (VP)-and catecholamine (CA)-containing nerve terminals, and their distribution and relationship were observed under electron microscopic level. The results showed that in the rat neurohypophysis there were not only widely distributed VP nerve terminals, but also there were many 6-OHDA induced degenerated nerve endings. The close relationship even synapse-like contacts existed between the CA-ergic endings and pituicytes as well as microglial cells. It was very interesting that the CA-ergic boutons formed axoaxonic synapses with VP-containing boutons. In this case, the CA-bouton was presynaptic element whereas the VP-bouton served as postsynaptic element. The above mentioned results probably provided ultrastructural evidence for the regulatory mechanism of the neurohormone releasing in the neurohypophysis for the first time.

Objective To investigate the effect of astrocytes activated by TNF? on the relapse of epilepsy. Methods The purified astrocytes of cultured rat hippocampus were stimulated by TNF?.The expression of NF\|? Bp65 was checked by immunocytochemical method and the release of glutamate(Glu) from astrocytes was measured by high performance liquid chromatography(HPLC).The astrocytes conditioned medium(ACM) stimulated by TNF? was collected and injected into lateral ventricle of chronic Coriaria Lactone kindled rats during the interictal period.The changes both in behavior and electroencephalogram(EEG) were recorded.The expression of NF\|? Bp65 and glial fibrillary acidic protein(GFAP) of hippocampus were also studied by immunocytochemical method. Results 1.TNF? could promote the release of Glu by astrocytes and quickly induce the nuclear expression of NF\|? Bp65 in cultured astrocytes.2.After lateral ventricular injection,four class epileptic behaviors were induced,which were confirmed by epileptic charge EEG.At 0\^5?h after injection of ACM,NF ? Bp65 expression in hippocampal cells especially in the CAl area were induced.It reached to the maximal levels at 2\|4?h and returned to control level at 8h.At 1h after injection of ACM,the increase of the number of positive GFAP \|immunoreactivity(IR) cells of hippocampus could be observed.The expression of GFAP reached to the top level at 4h and still maintained at a higher level than the control group until 8h.Conclusion\ Astrocytes activa\|ted by TNF? could induce the relapse of epilepsy by some solube neuro\|active substance.

Objective To explore the relationship between interleukin-1?(IL-1?) and group Ⅱ metabotropic glutamate receptors(mGluR2/3). Methods The rats were randomly divided into five groups:1^cotrol;2^group Glu;3^group IL-1?+Glu;4^group IL-1?;5^group IL-1?+D-AP5+Glu.The behavior of animals was observed.Expression of mGluR2/3 mRNA was assayed with RT-PCR. Results The alteration of behavior showed that the latent period of seizure in group with IL-1?+Glu was shortened significantly and the degree of seizure was also more serious as compared with that in group Glu.Expression of mGluR2/3 mRNA showed marked enhancement in group with IL-1?+Glu and group with Glu compared with control.However,expression of mGluR2 mRNA in group IL-1?+Glu was markedly decreased as compared to that in group Glu;the level of mGluR3 mRNA showed no apparent difference between group IL-1?+Glu and group Glu.Conclusion Present study suggests that IL-1? enhances the degree of seizure induced by glutamate,the mechanism may be due to its direct or indirect effects on down-regulation of expression of mGluR2 mRNA.

OBJECTIVE:To optimize the inclusion technology of Eugenol-β-cyclodextrin (β-CD) inclusion complex,and to identify and characterize it. METHODS:With the molar ratio of eugenol to β-CD,inclusion temperature and inclusion time as fac-tors,using the yield of inclusion compounds as index,the inclusion technology was optimized by orthogonal test. The formation of inclusion compound was identified by the spectra change of FT-IR,XRD and 1H NMR. Its structure was characterized by 1H RO-ESY NMR. RESULTS:The optimized inclusion conditions were that the molar ratio of eugenol to β-CD was 1.0:1;inclusion tem-perature was 60 ℃;inclusion time was 2.0 h. And the yield of inclusion compound was 73.86%(RSD=0.17%,n=3). 1H NMR results ofβ-CD and its inclusion complex indicated that the optimum qualitative ratio of the inclusion complex was 1.0:1. The inter-molecular interaction between eugenol and β-CD was confirmed by the spectrum analysis of FT-IR and XRD. 1H ROESY NMR re-sults indicated the structure of inclusion complex mainly was that the phenyl of eugenol was in the cavity of β-CD,the vinyl was outside. CONCLUSIONS:The inclusion technology is reasonable and feasible,and can be used for the inclusion of eugenol andβ-CD. The formation of inclusion compound is confirmed by the spectrum analysis.

The distribution of somatostatin (SRIF) in substantia gelatinosa of the rat spinal cord was studied by means of immuno-electron microscopy. The ultrastructural features showed that nerve terminals containing SRIF take part in forming presynap- tie elements of axe-somatic, axe-dendritic and axo-axonic synapses. The immune- reactive products locatl at the external membrane of mitochondria, around the small clear synaptie vesicles and in the large granular vesicles. Most of synaptic vesicles are round or ovoid in shape. Only a few of them are flattened. Based on the ultrastructural characteristics mentioned above and related experimental results the authors believe that SRIF in substantia gelatinosa of the rat spinal cord is probably involved as a neurotransmitter instead of neuromodulator.

The ultrastructural localization of peptide substances——VP,SP,ENK,SRIFand distribution of peptidergic nerves in rat neurohypophysis were studied by elect-ron microscopy.The results showed that VP,SP,ENK and SRIF immunoreactiveproducts located in large granular vesicles(110-150 nm in diameter),at the surfaceof microvesicles and outer membrane of mitochondria.The VP-,SP-,ENK- andSRIF-containing nerve fibers distributed at periphery of capillaries and vicinity ofpituicytes.VP- and ENK- positive nerve terminals formed axo-axonic synapses withnegative terminals.Axo-axonic synapses also existed between two SRIF-positive ter-minals.In addition,ENK- and SP-positive terminals formed synaptoid structureswith pituicytes.The authors have discovered the VP-positive pituicytes for the firsttime.The scientific significances of present paper are as follows:(1)Several kinds ofpeptide such as VP,SP,ENK and SRIF have been identified in neurohypophysis atultrastructural level;(2)The discovery of VP-positive pituicytes and their peptide-rgic innervation imply that these structures probably participate in the regulation ofneurohormone releasing;(3)New morphological bases have been provided for theregulation of neurohormone releasing.

6-OHDA was injected into the third cerebroventricle of rats.24 hours later,thanimals were sacrificed and the degeneration of catecholaminergic (CA) nerve term-inals were studied by electron microscopic cytochemical method.The results showedthat there were a number of degenerated nerve terminals in the arcuate nucleus.Thecharacteristics of these terminals were as follows:enhanced axoplasmic electron opa-sity,destroyed mitochondria and synaptic vesicles and the formation of dense bodies.The multilocular forms were commonly encountered.Most of degenerated terminalswere surrouded by processes of glial cells.The degnerated perikarya of the neuronsin arcuate nucleus with decreased RER,destroyed mitochondria and dense bodieswere also observed.Based on the characteristics of degenerated features the authorsbelieved that these damaged terminals should mainly belong to DA fibers.Theymight regulate or control the function of the arcuate nucleus by means of differentkinds of synaptic mechanisms.The source of DA nerve fibers may originate from A_(12)A_(14) or A_(15) cell groups of the hypothalamus.

To explore the effect of coriaria lactone (CL)-activated astrocyte-conditioned medium on the cerebral TNF-alpha of normal rats, the CL-activated astrocyte-conditioned medium (ACM) was injected into the lateral ventricle of SD rats. The rats were observed for behavioral changes, and the changes of the expression of TNF-alpha in the cerebral cortex and hippocampus were immunohistochemically examined by employing SP method. TNF-alpha level was assessed by means of radioimmunoassay in homogenate of cerebral cortex and hippocampus as well as cerebrospinal fluid. Seizure episodes were observed in ACM group 30 min after the ACM injection, but they were not observed in the control group. Immunohistochemical detection showed that the immunoreaction of TNF-alpha in hippocampus and cerebral cortex of rats were stronger than that of the control group 4 h after the ACM injection (P<0. 05). In this group, the concentrations of TNF-alpha in homogenate of cerebral cortex and hippocampus and cerebrospinal fluid were higher than those of the control group (P<0.05). It is suggested that the ACM activated by CL can enhance the expression of TNF-alpha in normal rats, and is related to epileptogenesis.