In our previous studies substance P-immunoreactive nerve fibers have been demonstrated in the pars distalis of the anterior pituitary in macaque monkeys, and they have been found to contact with somatotropes. The present study investigated the relationship of the substance P immunoreactive fibers to thyrotropes and corticotropes. Macaca mulatta monkeys were used. Sections of the anterior pituitary were double immunostained with antisera against either substance P and human thyroid stimulating hormone or substance P and human ACTH. Substance P immunoreactive varicosities were found to be in close proximity to thyrotropes and corticotropes. It is therefore suggested that a direct neural factor may take part in the regulation of TSH and ACTH secretion

The anterior pituitary gland of Macaca mulatta and M. assamensis were stained with antiserum against substance P. A substantial amount of substance P-immunoreactive (SP-ir) nerve fibers with numerous varicosities were found in the pars distalis of the anterior pituitary. They were located in the median part of the gland and were distributed mainly in its dorso-posterior region. The great majority of the varicosities were found to be related to the glandular tissue, although some apparently were located along the wall of the blood sinuses. Also many SP immuno reactive cells, mostly large and oval, and were distributed at the periphers of the gland. In areas where both SP-ir nerve fibers and cells were present, many cells were found to be in close proximity to nerve fibers.

A small amount of CGRP-ir nerve fibers were found immunohistochemically with an antiserum directed against rat CGRP in all parts of the pars distalis of adenohypophysis in the rat. They occurred in small patches or as scattered fibers. More fibers were seen in the median and dorsal parts of the gland than in lateral and ventral parts. Bundle of CGRP-ir never fibers were often observed within the pia mater covering the pituitary for some distance before entering the parenchyma. A few CGRP-ir cells were observed in the center of the caudal part of the adenohypophysis. None of them were found to be related to the CGRP-ir fibers. The large number of varicosities present along nerve fibers suggests that they may play a role in the regulation of the functions of the adenohypophysis.

WGA-HRP retrograde tracing technique combined with immunohistochemical method was used to study the source of somatostatin postitive fibers in the posterior pituitary. Retrogradely labeled somatostatin immunoreactive cells distributed mainly in the periventricular area from the level of the anterior to posterior magnocellular paraventricular nucleus; single double labeled cells were also found in the periventricular areas at the level of the anterior commissure and posterior fornical nucleus. A few double labeled cells were seen in medial parvocellular paraventricular nucleus and the medial part of posterior magnocelluar paraventricular nucleus. The double labeled cells in the periventricular area lie mainly beneath the ependymal layer. Some were seen to intercalated in-between the ependymal cells, bringing themselves very close to the cerebrospinal fluid, but no direct fluid contacing elements were verified.

The WGA-HRP retrograde tracing combined with immunohistochemical method was used to study the sources of CCK positive fibers in the male rat posterior pituitary. The CCK positive neurons projecting to the posterior pituitary localized mainly in the paraventricular nucleus, periventricular areas,and medial preoptic area. A few double labeled neurons were demonstrated in the subependymal area of the interventricular foramen and the floor of the 3rd ventricle. The CCK positive neurons projecting to the posterior pituitary, which weakly stained, were found in the bed nucleus of the stria terminalis, lateral hypothalamic area and the dorsal part of the supraoptic nucleus, too. Our result showed much more distribution of CCK-ir neurons projecting to the posterior pituitary in the hypothalamus than Palkovits′conclusion that the CCK positive fibers in posterior pituitary almost all originated from the magnocellular paraventricular nuclei. Some double labeled neurons in periventricular area were closely approximte to 3rd venticular cavity which suggested that these neurons may monitor the changes in the cerebrospinal fluid.

Objective To investigate the effect of all-trans retinoic acid on the change in intracellular glycogen in ovarian epithelioma cell line in vitro and ovarian epithelial carcinoma in nude mice in vivo. Methods COC2 cells were treated with all-trans retinoic acid in 1, 5, 10 and 30?mol/L drug concentrations for different length of time, and then intracellular glycogen and LDH were determined by biochemistry assay. Morphologic changes were observed with light and electron microscopy. CAOV3 tumor-bearing nude mice were treated with intragastric injection of the same drug in a dose of 2mg/(kg?d) for four weeks. The tumor samples were harvested thereafter for pathological study with histochemical and immunohistochemical staining, and also with electron microscopy. Results Intracellular glycogen was significantly increased, while LDH level was lowered after the cell line was treated with 5~10?mol/L of all-trans retinoic acid, and apoptosis of cancer cell occurred after using 30?mol/L of the drug. These changes were also observed in CAOV3 cells of tumor-bearing nude mice. Conclusion Our results suggest that treatment with all-trans retinoic acid resultin an increase in intracellular glycogen and decrease in LDH level both in COC2 cells in vitro and in CAOV3 tumor-bearing nude mice in vivo, and the suppression of tumor cell proliferation may be attributed to retarded intracellular metabolism.

Objective To evaluation of the result of transplantation of BDNF gene transfected olfactory ensheathing cells (OECs) to repair spinal cord injury (SCI). Methods The rat with SCI were divided into non-OECs, OECs and transfected OECs groups. The evidences of anatomical regeneration across the transection site were demonstrated by both neuronal tracing and immunohistochemical methods. Results 12w after implantation the retrograde labeling study with flurogold showed abundant labeled neurons in the superior part of CNS. There were a lot of labeled neurons in the intermediate zone and dorsal column of lower thoracic spinal cord rostral to the transection site. Labeled neurons were also observed in the upper thoracic and cervical spinal cord, reticular nucleus and raphe nucleus of oblongata, as well as reticular nucleus and parabrachial nucleus of pons. The anterograde labeling of PHA-L showed that regenerated fibers of upper neurons overcame glial scar and regrew into the distal part of spinal cord through the grafts. Some labeled fibers extended long enough to the caudal gray matter of the transected site. Conclusion The implantation of OECs transfected by BDNF gene may benefit the survival and regeneration of the injured axons, and accelerate the repair of the injured spinal cord in a more efficient way than that with OECs alone.

Objective To investigate the apoptosis rate of neurons in vitro among the primary culture cells isolated from rat spinal cords before and after injury. Methods The spinal cord neurons of Wistar rats at 14-day gestation were isolated and cultured. The neuronal processes were then injured by cutting. At different time points after injury, TUNEL method was employed to detect the apoptotic neurons. Results Before injury, there was almost no apoptotic neuron. However, a large amount of apoptotic neurons were observed after the injury. The highest incidence of apoptosis appeared on the first day, and then it gradually reduced in the following days, but increased in amount again on the seventh day. Conclusion The results of the present experiment reveal the regularity of apoptosis of neurons before and after injury, and it provides a platform for further research regarding therapeutic intervention in the treatment of spinal cord injury.

Objective To establish a model of injury to primarily cultured spinal cord neurons,mimicking the neuronal injury after complete transactional spinal cord trauma,for the sake of exploring changes in expression of an immediately-early gene,c-jun,in central nervous system injury. Methods Spinal cords were removed form fetal Wistar rats at the 14th gestation day and the neurons were cultured for 10 to 12 days.Then,mechanical injury was applied to the neurons by making regular scores on the culture disk under direct vision with the aid of a self-made standard template.Morphology of the injured neurons and changes in expression of c-Jun protein were observed before and at different intervals after injury. Results c-Jun expression was noted in neuronal nuclei 10 min after injury and its peak appeared at 2 hrs.Besides,the density of positive neurons bore evidently an inverse proportion with their distance from the scores.Conclusion\ Positive expression in injury neurons show that c-jun gene enters the nucleoli of injured neurons and takes the role of “the third messenger” at early time after neuronal injury.

OBJECTIVE:To find the active ingredient of on antithrombotic chuanxiong rhizoma using computer aided drug de-sign. METHODS:Usingthrombosisas keyword,thrombosis related proteins were searched and screened in therapeutic target da-tabase;target proteins'three-dimensional structure were downloaded in protein database,then the protein preparing tool were used to determine the coordinates of the active area center. PyRx software and Discovery Studio Visualizer were used to match the 247 small molecules of chuanxiong rhizoma with target protein that downloaded from Taiwan traditional Chinese medicine database. The active molecules were screened and binding force was analyzed. RESULTS:Active molecules of neochlorogenic acid,1-H-benz-imidazole-2-amine,3,8-dihydrodiligustilide,chuanxiongterpene were selected by blinding energy,and there were high binding ac-tivity among these active molecules,thrombin,antithrombinⅢ,coagulation factorⅩa and thrombomodulin,and the binding ener-gy were -6.1,-4.5,-7.7,-8.6 kJ/mol. Analysis results showed van Edward force and electrostatic interactions played an im-portant role in their respective docking. CONCLUSIONS:Neochlorogenic acid,1-H-benzimidazole-2-amine,3,8-dihydrodiligusti-lide,chuanxiongterpene may be the antithrombotic activity ingredients of Chuanxiong rhizoma.