Discussion on the construction problems and countermeasures of hospital scientific research management subsystem based on HRP system / 中华医学科研管理杂志

Objective This paper studied the characteristics and contents of the hospital scientific research management subsystem based on HRP system,analyzing its characteristics and functions,proposing solutions to identified problems to improve the hospital management and core competitiveness.Methods Through the analysis of the characteristics and design module contents of the research management subsystem of HRP system,some problems were identified in the system,such as the long adaptation process of researchers' new or old concepts,unstable research management subsystem affecting the links between departments and so on.Results To better deal with identified problems,this paper put forward some improvement measurements,such as strengthening the monitoring of H RP system's scientific research management subsystem,strengthening the safety construction of both HRP system and scientific research management subsystem.Conclusions HRP system and scientific research management subsystem reflect the refinement,scientific and integration of hospital management,which is a hospital management concept and a means to improve the core competitiveness of the hospital.We should change the percep tions,improve understandings,strengthen the collaboration,and promote the overall development of the hospital.

Preparation and identification of rabbit anti-marmoset IgG-HRP conjugate antibody / 中国比较医学杂志

Objective To purify marmoset serum IgG, prepare and identify the antiserum and the rabbit anti-marmoset antibody IgG-HRP (horseradish peroxidase). Methods Using SDS-PAGE analysis to identify the serum IgG from HiTrapTM Protein G. The antiserum titer was determined by double immunodiffusion assay. The rabbit anti-marmoset IgG was labeled with HRP by improved sodium periodate method. ELISA and western blotting were used to evaluate the concentration and specificity of rabbit anti-marmoset IgG-HRP. Results The purity of purified marmoset serum IgG determined by SDS-PAGE was higher than 95% , and the anti-serum titer of the anti-marmoset IgG polyclonal antibody was 1∶64. The concentration of rabbit anti-marmoset IgG-HRP identified by direct ELISA was 1∶256 000, and that by western-blotting was 1∶15 000, with a strong specificity. Conclusions The IgG-HRP marker antibody is prepared and the specificity and concentration are identified by ELISA and western blotting. It reserves the resources for the detection system of marmoset pathogens and the molecular immunological testing system.

Effect of Tinospora cordifolia (Guduchi) on the phagocytic and pinocytic activity of murine macrophages in vitro.

Tinospora cordifolia (Guduchi) is a widely used herb in Ayurvedic system of medicine known to possess immunomodulatory properties. The present study was aimed to study the activation of macrophages after in vitro guduchi treatment. The aqueous extract of T. cordifolia was found to enhance phagocytosis and pinocytosis in vitro. The rate of pinocytosis by macrophages when measured by uptake of horseradish peroxidase was significantly increased after guduchi treatment as compared to medium alone. The macrophages demonstrated an increased phagocytosis to non-infective microorganisms (heat killed yeast) and live infective microorganisms (E. coli) after guduchi treatment. The results demonstrate that Guduchi enhances macrophage activation as analyzed by cytochemical parameters.

Analysis of polymorphism of Pfhrp2 gene in Plasmodium falciparum from falciparum malaria patients in Yunnan Province / 中国血吸虫病防治杂志

Objective To analyze the polymorphism of histidine rich protein 2(HRP II)gene in Plasmodium falciparum (Pfhrp2)from falciparum malaria patients in Yunnan Province,so as to lay the foundation for studying the defection of antigen genes of Plasmodium. Methods The filter paper blood samples and related information of falciparum malaria cases reported were obtained in Yunnan Province from August 2012 to September 2015. Under the guidance of the specific primers,the exon2 regions in Pfhrp2 gene in P. falciparum from DNA samples were amplified by PCR,and the PCR products were sequenced. The sequences of exon2 region in Pfhrp2 gene were blasted by comparing with the reference sequences AY816237,AY816240,and AY816301. Next,the polymorphism of the sequence in exon2 region of Pfhrp2 gene was analyzed by MEGA 5.04 software. The conserved sites and genetic distances between sequences were calculated by using the software as well,and the clustering tree was drawn according to the genetic distances between the amino acid sequences. Results A total of 218 bloods samples from the falciparum malaria cases in 15 prefectures of Yunnan Province were collected,and the sources of infection included Yun?nan,Africa and Myanmar. The PCR results showed that the exon2 regions in Pfhrp2 genes of 155 samples were positive by am?plification and their products were sequenced successfully. The sequence analysis showed that the length range of the amino acid residues of exon2 region in Pfhrp2 gene was from 115 aa to 298 aa,the average length was 239.7 aa. There was no statistically significance among the means of the amino acid residues of the isolates from Africa( 239.9 aa),Myanmar(239.5 aa)and Yun?nan(241.6 aa)(F=0.025,P>0.05). All the 155 amino acid sequences ended with type 12 repeat,98.1%(152/155)of them started with type 1 repeat and 1.9%(3/155)of them started with type 2. Type 2 presented most frequently repeat in all the se?quences and the average repeat times were 12.9. The homologous locus of the DNA sequences in exon2 regions of the 155 Pfhrp2 genes was 894 bp,among which the conservative sites accounted for 20.6%(186/894),and the variable sites for 78.2%(699/894). The genetic distances between the sequences of Africa isolates ranged from 0 to 0.741,and those of the Myanmar and Yun?nan isolates were 0-0.948 and 0-0.750,respectively. The cluster analysis showed that all the 155 sequences clustered into 3 cat?egories on genetic distances between amino acid sequences according to the size of the amino acid sequence length. At the same level,the sequences had approximate lengths and amino acid repeat types. Conclusion The sequence of exon2 region in Pfhrp2 gene of P. falciparum from falciparum malaria cases in Yunnan Province is highly polymorphic,the P. falciparum iso?lates are clustered mainly according to the size of the amino acid sequence of exon2 region in Pfhrp2 gene.

Assessing the Performance Characteristics of the “CareStartTM Malaria HRP2 pf (CAT NO: G0141, ACCESSBIO)” Rapid Diagnostic Test for Asymptomatic Malaria in Mutengene, Cameroon.

Aim: The aim of this study was to determine the prevalence and density of malaria parasites in asymptomatic school children in Mutengene and evaluate the performance characteristics of the ‘CareStartTM Malaria HRP2 pf (CAT NO: G0141, ACCESSBIO)’ rapid diagnostic test (RDT) using light microscopy as a gold standard. Study Design: The study was a cross-sectional survey. Place and Duration of Study: The study was carried out in Mutengene, from February to March, 2013. Methodology: A total of 406 pupils were studied. Demographic data was taken for each child and capillary blood was collected. Blood films were prepared for the assessment of parasite density and speciation. A drop of blood was used on the RDT to determine the malaria status. Results: The mean age at 95% confidence interval (CI) was 8 ± 2 years (range = 4 -15 years) and the overall prevalence of malaria was 39.9% (162) by microscopy. The geometric mean parasite density (GMPD) was 2332.7 parasites/µL (range: 218 - 16000). Only 386 pupils were examined by both methods. More pupils were positive by microscopy (40.9%, CI = 36.1 - 45.9) than by RDT (27.9%, CI = 23.7 - 32.7) and the difference was statistically significant (χ2 = 16.1, P <0.0001). The majority of those detected had high infection (≥ 5000 parasite/µL). Less than 50% of those with low (25.0%, CI = 12.0 - 44.9), moderate (40.7%, CI = 32.24-49.70) and high parasitaemia (75%, CI = 5.00-89.82) were positive by RDT and the difference was significant (χ2 = 10.09, P = 0.006). The RDT showed a low sensitivity of 48.5% (CI = 40.3 – 56.9%) and specificity of 84.0% (CI = 80.0- 88.2%). Conclusion: More research needs to be done on the RDT to improve on its performance characteristics before it could be used in mass surveillance programmes.

The brainstem localization of gastric preganglionic parasympathetic neurons in the Agouti (Dasyprocta leporina) / Localización del tronco encefálico de las neuronas parasimpáticas preganglionares gástricas en el agutí (Dasyprocta leporina)

This study was designed to determine qualitatively, the source of gastric vagal nerve fibres in the Agouti. A total of 18 male and female adult agoutis were used for the present investigation. Following anaesthesia, laparotomy was performed and the stomach exteriorized. Multiple intramuscular injections of wheat germ agglutinin-horseradish peroxidase (WGA-HRP) were then made into different areas of the stomach in the experimental animals. The control animals were divided into four groups of two animals each. The first group had intraperitoneal injection of the tracer, the second had intramuscular injection of normal saline, the third group had injection of tracer into the hepatic portal vein and the last group had injection of the tracer into the gastric walls followed immediately by bilateral vagotomy. Following a survival period offive to seven days, the animals were sacrificed by transcardial perfusion, first with normal saline followed by fixative and finally with 20% buffered sucrose. Following perfusion, the brainstem was extracted from the brain, immersed in 20% buffered sucrose and kept refrigerated overnight for cryoprotection. The brainstems were subsequently sectioned serially, processed for WGA-HRP neurohistochemistry and then analysed under light and dark-field illuminations. The analysis of the sections taken from the experimental animals revealed bilateral presence of WGA-HRP labelled neurons in the dorsal motor nucleus of the vagus nerve (DMNV) and the nucleus ambiguus (nA) of the medulla oblongata. No labelled neurons were seen in any of the sections taken from the control animals. The implications of the findings are discussed.

Este estudio fue diseñado para determinar cualitativamente el origen de las fibras gástricas del nervio vago en el agutí. Un total de 18 agutíes adultos masculinos y femeninos fueron utilizados para la presente investigación. Después de la anestesia, se realizó una laparotomía y se sacó el estómago al exterior. Luego se hicieron múltiples inyecciones intramusculares de aglutinina de germen de trigo con peroxidasa de rábano (WGA-HRP) en diferentes áreas del estómago de los animales experimentales. Los animales del control fueron divididos en cuatro grupos de dos animales cada uno. Al primer grupo se le puso una inyección intraperitoneal del marcador; al segundo se le administró una inyección intramuscular de solución salina normal; al tercer grupo se le inyectó el marcador en la vena porta hepática; y al último grupo se le puso la inyección del marcador en las paredes gástricas, seguida inmediatamente por una vagotomía bilateral. Tras un periodo de supervivencia de cinco a siete días, los animales fueron sacrificados por perfusión transcardíaca, primero con solución salina normal, seguida de fijador, y finalmente con sacarosa tamponada al 20%. Después de la perfusión, el tronco encefálico fue extraído del cerebro, inmerso en sacarosa tamponada al 20%, y mantenido en refrigeración durante la noche para su crioprotección. Los tronos encefálicos fueron luego seccionados en serie, procesados para para el análisis neuro-histoquímico mediante aglutinina de germen de trigo con peroxidasa de rábano, y analizados entonces bajo iluminaciones de campo de luz y campo oscuro. El análisis de las secciones tomadas de animales experimentales reveló la presencia bilateral de neuronas etiquetadas WGA-HRP en el núcleo motor dorsal del nervio vago (DMNV) y en el núcleo ambiguo (nA) de la médula oblonga. No se observaron neuronas etiquetadas en ninguna de las secciones tomadas de los animales de control. Se discuten las implicaciones de los hallazgos.

Detrimental effect of electromagnetic pulse exposure on permeability of in vitro blood-brain-barrier model / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To study the effect of electromagnetic pulse (EMP) exposure on permeability of in vitro blood-brain-barrier (BBB) model.</p><p><b>METHODS</b>An in vitro BBB model, established by co-culturing brain microvascular endothelial cells (BMVEC) and astroglial cells (AC) isolated from rat brain, was exposed to EMP at 100 kV/m and 400 kV/m, respectively. Permeability of the model was assayed by measuring the transendothelial electrical resistance (TEER) and the horseradish peroxidase (HRP) transmission at different time points. Levels of BBB tight junction-related proteins were measured at 0, 1, 2, 4, 8, 12, 16, 20, 24 h after EMP exposure by Western blotting.</p><p><b>RESULTS</b>The TEER level was lower in BBB model group than in control group at 12 h after EMP, exposure which returned to its normal level at 24 h. The 24 h recovery process was triphasic and biphasic respectively after EMP exposure at 100 kV/m and 400 kV/m. Following exposure to 400 kV/m EMP, the HRP permeability increased at 1-12 h and returned to its normal level at 24 h. Western blotting showed that the claudin-5 and ZO-1 protein levels were changed after EMP exposure.</p><p><b>CONCLUSION</b>EMP exposure at 100 kV/m and 400 kV/m can increase the permeability of in vitro BBB model and BBB tight junction-related proteins such as ZO-1 and claudin-5 may change EMP-induced BBB permeability.</p>

New fluorimetric assay of horseradish peroxidase using sesamol as substrate and its application to EIA / 药物分析学报

Horseradish peroxidase (HRP) is generally used as a label enzyme in enzyme immunoassay (EIA).The procedure used for HRP detection in EIA is critical for sensitivity and precision.This paper describes a novel fluorimetric assay for horseradish peroxidase (HRP) using sesamol as substrate.The principle of the assay is as follow:sesamol (3,4-methylenedioxy phenol) is reacted enzymatically in the presence of hydrogen peroxide to produce dimeric sesamol.The dimer is fluorescent and can be detected sensitively at ex.347 nm,em.427 nm.The measurable range of HRP was 1.0 × 10-18 to 1.0 × 10-15 mol/assay,with a detection limit of 1.0 × 10-18 tmol/assay.The coefficient of variation (CV,n=8) was examined at each point on the standard curve,with a mean CV percentage of 3.8％.This assay system was applied to thyroid stimulating hormone (TSH) EIA using HRP as the label enzyme.

Segmental and Regional Innervation of Acupoint "Taixi" (KI 3) Area in the Rat: A Horseradish Peroxidase Method Study / 针刺研究

L6.A longitudinal column of HRP-labeled motoneurons was found in the dorsolateral and mediolateral portions of the spinal cord,distributing in the lamina Ⅸ from the caudal L4 to the rostral L6.Additionally,the transganglionic HRP-labeled central projection axonal terminals were found to be dense in the central part of laminae Ⅰ-Ⅱ from L4 to the rostral L6,and to scatter in the central part of gracile nucleus.Conclusion HRP-labeled primary afferent and efferent innervating acupoint "Taixi"(KI 3) are DRGs of L4-L6,the dorsolateral and mediolateral motoneuron columns of L4-L6,and the centrally projecting axonal terminals of laminae Ⅰ-Ⅱ of the spinal cord and the gracile nucleus.

Morphological Changes of the Sensory Neurons in the Peripheral Neuropathy of Rat Tibial Nerve Using WGA-HRP Tracing Method / 체질인류학회지

Neuropathy is a general term referring to disorders of nerves, and produces when the nerves are damaged. It is characterized by spontaneous pain, allodynia and hyperalgesia. The purpose of present study is to observe the number of WGA-HRP (wheat germ agglutinin-horseradish peroxidase) labelded sensory neurons of DRG (dorsal root ganglia), and distributions according to cell size of sensory neuron in tibial nerve ligation model (NLM). The tibial nerve ligation was performed with 3-0 silk by the application of three tight ligatures at the mid-thigh level. In the neuropathy model of rat tibial nerve ligation, morphological changes of sensory neurons in DRG were observed using WGA-HRP. Rats of NLM showed the neuropathic behaviors. Rats were shown guarding affected limb and limping. Their toes and ankle joint of operated limb were hyperflexed. Under light microscopy, tibial nerve showed degeneration of axons in NLM. In control and NLM, labeled sensory neurons of tibial nerve distributed L4 and L5 DRG. In control group, the labeled sensory neurons were round or oval in shape. They were large and small cells, and mixed pattern. Total number of labeled sensory neurons in NLM decreased significantly from control group. The number of labeled sensory neurons in L4 and L5 DRG decreased significantly from control group. Labeled large and small cells decreased significantly from control group. Present study may serve as the basic information about the changes of DRG sensory neurons in NLM.

Development of Enzyme Linked Immunosorbent Assay for Erythropoietin / 대한진단검사의학회지

BACKGROUND: The aim of our study was to optimize and establish erythropoietin (EPO) enzyme linked immunosorbent assay (ELISA) system. METHODS: We prepared several monoclonal and polyclonal antibodies specific to human-EPO. The best combinations of antibodies for coating and detecting antibodies were selected for the establishment of ELISA. We tested several methods such as a competitive EIA and a sandwich ELISA. RESULTS: The best sandwich ELISA was optimized compared to competitive EIA when purified polyclonal antibody (PoAb) was used as a coating antibody and biotinylated PoAb as a detecting antibody. This sandwich ELISA easily detected EPO when PoAb pairs were used compared to the ELISA using monoclonal antibody and PoAb. There were no significant differences between the effects of various blocking solutions on the performance of sandwich ELISA using biotinylated antibody. The ELISA system using PBST containing 3% BSA as a blocking solution can sensitively detect EPO (10 mU/mL) in a broad range of EPO concentrations (10-2,000 mU/mL) and there were cross-reactions with other cytokines). CONCLUSIONS: EPO can be easily determined by using biotinylated PoAb as a detecting antibody and another PoAb as a coating antibody.

Projections from respiratory neurons of medulla oblongata to the motor neuron pool of phrenic nerve in rat:An HRP retrograde tracing study / 中国康复理论与实践

@#ObjectiveTo observe the projections from respiratory neurons of medulla oblongata to the motor neuron pool of phrenic nerve in rat. MethodsBy using horseradish peroxidase (HRP) retrograde tracing technique after injecting HRP into phrenic nerve, retrograde labelled neurons were found in C3-C5 segment. Then, HRP was injected into the area where the phrenic motor neurons occupied, retrograde labelled neurons were found in medulla oblongata. ResultsPhrenic motor neurons locate in the C3-C5 segment ipsilaterally, occuping the intermediate portion of anterior horn and appearing typical motor neurons. Retrograde labelled neurons were found bilaterally in medulla oblongata, the neurons were located in nucleus retroambigualis (RNA), ventramedial area to nucleus retrofacialis (NRF). ConclusionThe phrenic motor neurons may receive direct projection of respiratory neuron in medulla oblongata, the projecting neurons are distributed in RNA and NRF area.

Quantitative Ultrastructural Analysis of Periodontal Afferent Terminals in the Trigeminal Motor Nucleus / 대한해부학회지

Little is known about processing mechanism of sensory input from the periodontal ligaments to the trigeminal motor nucleus for the control of chewing force and modulation of chewing pattern. Low threshold mechanoreceptive periodontal afferent was labeled with horseradish peroxidase by use of intra-axonal injection technique and investigated with electron microscopy. Quantitative ultrastructural analysis was performed on the 39 serially reconstructed labeled boutons in the trigeminal motor nucleus in cat. Labeled bouton contained clear spherical vesicles and one or two large dense cored vesicles. Most of labeled boutons were dome or round shape. All the analysed labeled boutons were presynaptic to dendritic shaft or distal dendrite and those presynaptic to soma or proximal dendrite were not observed. A large number of labeled boutons (46.2%) were postsynaptic to one or two presynaptic pleomorphic vesicle containing endings. Synaptic triad, in that a presynaptic ending which is presynaptic to the labeled bouton, in turn, is presynaptic to dendrite that is postsynaptic to the labeled bouton, was observed in 10.3% of the labeled boutons. Most of the labeled boutons showed simple synaptic organization, in that 64.1% of the labeled boutons made synaptic contacts with one or two neuronal profiles. One (2.6%) of the 39 analyzed labeled boutons showed synaptic contacts with 5 or more neuronal profiles. Labeled bouton volume, mitochondrial volume, apposed surface area and active zone area showed wide variation. These ultrastructural parameters were positively correlated with bouton volume. The values for apposed surface area and active zone area with presynaptic p-endings, in contrast to those with postsynaptic dendrites, showed narrow range and had little correlation with bouton volume. The present study revealed characteristic features on ultrastructural parameters of labeled boutons from periodontal afferent which is involved in periodontal masseteric reflex, and that influence on the postsynaptic trigeminal motoneurons showed wide variability.

Study on afferent projections from the abducens nucleus to the contralateral medial rectus subnucleus of the oculomotor nucleus in the rat / 重庆医科大学学报

Objective: To investigate afferent projections from the abducens nucleus to the contralateral medial rectus subnucleus of the oculomotor nucleus in rats.Methods:The means of retrograde hoseradish peroxidase tracing was used.Results:The internuclear neurons in the abducens nucleus were divided into two subgroups,the ventromedial and the ventrolateral ones. The axons of the internuclear nurons crossed the midline,ascended in the contralateral medial longitudinal fasciculus and intermediated exclusively in the contralateral medial rectus subgroups.Conclusion:The results imply that the internuclear meurons could play a major role in conjugate horizontal eye movement.

A Study on the Spinoreticulocerebellar Tract in Chickens

The spicoreticulocerebellar (SRC) tract is an indirect spinocerebellar tract formed by the reticular formation (RF), which is connected to the cerebellum and spinal cord. The RF receives ascending fibers to both the spinal enlargement and sends descending fibers to the cerebellum. This study demonstrated that the connectivity of the neurons in the RF is concerned to the cerebellum and spinal cord using the anterograde projection with biotinylated dextran amine (BDA) and retrograde labeling with wheat germ agglutinin-horseradish peroxidase (WGA-HRP). Until now, a preliminary study in mammals has dealt with the afferent and efferent pathways in separating groups of neurons in the RF. There are only few reports on chickens. This study examined the SRC tract in chickens. Following bilateral injections we injected BDA into chicken spinal cord (lumbosacral enlargement) and WGA-HRP into the cerebellum. Both of single- and double-labeled cells were found within the RF. The spinoreticular axons were mainly distributed from the potomedullary junction to the rostral medulla in the rostro-caudally RF levels, for example, nucleus of reticularis (n. r.) pontis oralis, locus coeruleus, n. r. pontis caudalis, n. r. pars gigantocellularis, n. r. gigantocellularis and n. r. parvocellualris. Reticulocerebellar labeling by the WGA- HRP was found in the same place as well as that of the BDA-projection. We observed that the proportion and location of double labeling cells in the chicken were almost similar in each level, comparing to the rodents. These results suggest that the reticular formation is strongly related to the spicoreticulocerebellar tract in chickens.

CHANGES OF Aβ FIBERS SPROUTING IN DORSAL HORN OF LUMBAR CORD ON NEUROPATHIC PAIN MODEL IN RATS / 神经解剖学杂志

It is proposed that following peripheral nerve injury abnormal sprouting of Aβ fiber primary afferent neurons in the spinal cord contributes to the allodynia that often occurs with such injury. The present investigation is to determine whether this sprouting is reversal after compression of peripheral nerve was relieved. In a rat model of neuropathic pain made by rat sciatic nerve compression,chorela toxin B subunit conjugated horseradish peroxidase (CB-HRP) was used to trace the termination of Afiber primary afferents and sections were reacted for using tetramethylbenzidine (TMB) as the chromagen. We demonstrated that the compression to the sciatic nerve also results in hyperalgesia and novel transganglionic CB-HRP staining in laminae Ⅱ, and this sprouting can not be reversed by decompression. This structural reorganization in central nervous system and its irreversible character may contribute to the development and maintenance of neuropathic pain.

Increased Labelling of Perivascular Phagocytes by Blood Borne HRP in the SAM P10 Brain / 대한해부학회지

The present study examined the uptake of blood borne horseradish peroxidase (HRP) by perivascular cells in the senescence -accelerated mouse prone -10 (SAMP10) and the senescence -accelerated mouse resistant -1 (SAMR1) brains. SAMP10 and SAMR1 brains were studied from mice of each of the following ages: 3 months old SAMP10, 12 ~14 months old SAMP10, 3 months old SAMR1, and 12 ~14 months old SAMR1. Animals were injected via a tail vein with HRP (type VI) solution. Two hours later animals were transcardially perfused with 4% paraformaldehyde and 1% glutaraldehyde mixture. After sectioning with a vibrating microtome, brain sections were stained using a DAB solution. Products of the HRP -DAB reaction were frequently and intensely labelled in the perivascular cells along the microvasculature, especially in the young SAMP10 brain. Electron microscopy revealed that the reaction products were evident in the endothelium of the microvasculature, as well as the perivascular phagocytes of arterioles or venules. In the aged SAMP10, the number of perivascular cells showing HRP -reaction products was lower than in the young SAMP10. Under the electron microscope, the perivascular cells of the aged SAMP10 brain showed very weak intensity of HRP staining, and these cells contained abundant foamy vacuoles or lipid droplets. In both young and aged SAMR1 brains, labelled perivascular phagocytes were very occasionally found. In summary, the present results showed increased uptake of blood -borne HRP by perivascular phagocytes in the young SAMP10 brain, and the age related decrease of this labelling, which suggests altered microvascular barrier function with aging in the SAMP10 brain.

Neural Pathway Innervating Epididymis of Rats by Pseudorabies virus (PRV-Ba-Gal) and WGA-HRP / 대한해부학회지

This experimental studies was to investigate the location of PNS and CNS labeled neurons following injection of 2% WGA-HRP and pseudorabies virus (PRV), beta-galactosidase inserted Bartha strain, into the epididymis of rats. After survival times 4~5 days following injection of 2% WGA-HRP and PRV-Ba-Gal, the rats were perfused, and their brain, spinal cord, sympathetic ganglia and spinal ganglia were frozen sectioned (30 mm). These sections were stained by HRP histochemical and beta-galactosidase histochemical staining methods, and observed with light microscope. The results were as follows : 1. The WGA-HRP labeled sympathetic ganglia projecting to the epididymis were observed in pelvic ganglion and L1-6 lumbar sympathetic ganglia. 2. The WGA-HRP labeled spinal ganglia projecting to the epididymis were observed in L1-6 spinal ganglia. 3. The beta-galactosidase labeled neurons projecting to the epididymis were observed in lamina VII of cervical segments. In thoracic segments, beta-galactosidase labeled neurons were observed in dorsomedial part of lamina I, II and III. Dense labeled neurons were observed in intermediolateral n. and dorsal commissural n.. In lumbar segment, labeled neurons were observed in lamina III, IV, V, dorsal commisural n. and superficial dorsal horn. 4. In the medulla oblongata, beta-galactosidase labeled neurons projecting to the epididymis were observed in the trigeminal spinal n., A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular n., rostroventrolateral reticular n., area postrema, n. tractus solitarius, raphe obscurus n., raphe pallidus n., raphe magnus n., parapyra-midal n., lateral reticular n. and lateral paragigantocellular reticular n.. 5. In the pons, labeled neurons were observed in Kolliker-Fuse n., locus coeruleus, subcoeruleus n. and A5 noradrenalin cells. 6. In midbrain, labeled neurons were observed in periaqueductal gray substance, retrorubral n., substantia nigra and dorsal raphe n.. 7. In the diencephalon, labeled neurons were observed in paraventricular hypothalamic n., lateral hypothalamic nucleus., medial preoptic n. and retrochiasmatic n.. These results suggest that WGA-HRP labeled neurons of the spinal cord projecting to the rat epididymis might be the first-order neurons related to the viscero-somatic sensory and sympathetic postganglionic neurons, and beta-galactosidase labeled neurons of the brain and spinal cord may be the second and third-order neurons response to the movement of vascular smooth muscle in epididymis. These beta-galactosidase labeled neurons may be central autonomic center related to the integration and modulation of reflex control linked to the sensory and motor system monitoring the internal environment. These observations provide evidence for previously unknown projections from epididymis to spinal cord and brain which may be play an important neuroanatomical basic evidence in the regulation of epididymal function.

Ultrastructural Analysis of GABA- and Glycine-Immunoreactive Nerve Terminals on Jaw-Closing and Jaw-Opening Motoneurons in the Rat / 대한해부학회지

Previous studies have shown that inhibitory synaptic inputs are different between in spinal and trigeminal motor systems and activities of jaw closing and opening alpha motoneurons are different during a chewing cycle. This study examined the distribution of inhibitory synapses made on masseter and digastric motoneurons by using retrograde tracing of wheat germ agglutinin conjugated to horseradish peroxides (WGA-HRP) combined with postembedding immunogold labeling on serial ultrathin sections.Many boutons IR (immunoreactive) to GABA and/or glycine were found to appose on two kinds of motoneurons, which were containing pleomorphic vesicles (a mixture of round, oval and flattened vesicles) and exhibited symmetrical synaptic contacts on the somata. Packing density and synaptic covering % were higher in digastric than in masseter motoneurons. Of 703 boutons apposing on 12 masseter motoneurons, 6.08+/-3.51, 29.67+/-8.89 and 17.78+/-5.22% were IR to GABA only, glycine only, and both GABA and glycine, respectively. Of 637 boutons apposing on 11 digastric motoneurons, 6.37+/-4.64, 19.74+/-8.25 and 12.01+/-5.38% were IR to GABA only, glycine only, and both GABA and glycine, respectively. Proportions of glycine IR boutons were higher than that of GABA IR boutons in both masseter and digastric motoneurons. Packing density and proportion of boutons IR to GABA and/or glycine were higher in jaw closing than in jaw opening motoneurons (packing density, 12.03+/-1.58 vs 10.28+/-2.99; proportion of IR boutons, 53.54+/-8.94% vs 38.12+/-9.38% in jaw closing and opening motoneurons, respectively). These results provide ultrastructural evidence that GABA and glycine act as important neurotransmitters for modulation of jaw movement and that proportion of inhibitory synapses is higher in jaw closing than in jaw opening motoneurons.

Experimental study of evaluation nerve regeneration with ~ 125 Iodine-horseradish paroxidase after operation / 中华显微外科杂志

Objective To establish a experimental method for evaluating nerve regeneration after operation by means of tracing the retrograde axonal plasm flow by 125 I HRP Methods Eighteen SD rats were used The right sciatic nerve of mouse was transected at the site 0 8cm distal to sciatic notch and then anastomosed immediately Four weeks later,500?l of 125 I HRP solution was injected into the right musculi triceps surae The right sciatic nerve segment proximal to the anastomosis site was cut for checking up the intensity of radioactivity in each tissue at 24 hours after injection The fiber number of nerve distal to the site of anastomosis were calculated The relationship between the intensity of radioactivity and the fiber number was analysed Results The relationship between the intensity of radioactivity of sciatic nerve segment proximal to the site of anastomosis and the fiber number of distal sciatic nerve segment was correlation and there was statistic significance. Conclusion The status of nerve regeneration distal to the site of anastomosis can be evaluated by means of intensity of 125 Iodine in the nerve segment proximal to the site of anastomosis