Projections from substance P (SP)-and cholecystokinin (CCK) containing neurons in the periaqueductal gray(PAG)and Edinger-Westphal(E-W)nucleus to the spinal cord were studied by means of the combining method of HRP tracing with immunocytochemistry in rats. The results showed that a few neurons in the ventrolateral region of PAG projected bilaterally to the cervical, thoracic and lumbar segments of the spinal cord, with the predominant projections from the ipsilateral side. The authors reported that these descending projection neurons showed SP-like immunoreactivity for the first time(account for 48%). The neurons of E-W nucleus projected diffusely to all segments of the spinal cord contained SP (70%) or CCK (73%) respectively, suggesting that at least a part of E-W neurons projecting to the spinal cord contain both SP and CCK.

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.

Using immunoelectron microscopic technique the SPergic autoregulatory synapses in the nucleus raphe magnus were studied. The results showed that there were SPlike perikarya and nerve fibers. The positive perikarya mainly were large polypolar cells. The positive fibers appeared as beaded-like and formed axodendritic synapses with SP-positive dendrites besides synapses formed with SP-negative structures. In the autoregulatory synapses the pre- and postsynaptic element were both labeled with immunoreactive products which precipitated at the periphery of small clear synaptic vesicles and the dendritic tubes, on the outer membrane of mitochondria, and in the cytoplasmic matrix. The pre- and postsynaptic membrane were symmetrical, and the synaptic vesicles aggregated near the presynaptic membrane. The synaptic cleft was about 20 nm in width and contained electron dense materials. The generality, structural characteristics and functional significance of the autoregulatory sysnapses were discussed.

The relationship between CCK-and VP-neurons in the rat retrochiasmatic area was studied at ultrastructural level by means of pre-embedding(PAP) double immunoelectron microscopic labeling technique. First, the VP-immunoreactivity was demonstrated by DAB method. After thoroughly washing, the CCK-immuno- reactivity was revealed by ammonium molybdate-TMB method. Being stabili- zed by DAB-cobalt chloride, the sections were embedded in Epon 812. Under ele- ctron microscope, it was observed that in the retrochiasmatic area, the VP-LI products distributed diffusely as high electron dense granular or flocculent depo- sits, whereas the CCK-LI products distributed sparsely as needle-or mass-like deposits. VP-LI perikarya were small in size with oval shape and CCK-LI peri- karya were medium in size with polygonal shape. CCK-LI perikarya and dendri- tes received afferent synapses from non-CCK- and non-VP-axonal terminals VP- LI axons received afferent synapses from VP and non-VP-axonal terminals It was interesting that the VP-LI axonal terminals formed efferent axoaxonic syna- pses with CCK-LI axonal endings and, vice versa, the CCK-LI axonal terminals established also efferent axoaxonic synapses with VP-LI axonal endings. The above mentioned results identified for the first time that in the rat retrochiasma- tic area not only there were CCK- and VP-neurons, but also there were reciprocal synaptic regulations between above two kinds of peptidergic neuron, providing new ultrastructural basis for the regulatory mechanism of the neuroendocrine in hypothalamus.

Using electron immunocytochemical method, the ultrastructural distribution and the synaptic connections of CCK-containing neurons in the paraventricular nucleus (PVN) of the rat were studied. The results showed that the CCK-like immunoreactive products located in farge granular vesicles, cytoplasmic matrix, at the periphery of small clear vesicles, rough endoplasmic reticulum and the membrane of mitochondria. The CCK-positive nerve cell bodies were large or small in size and distributed mainly in the medial part of the PVN, subependymal region and the vicinity of capillaries. Some of them as postsynaptic elements formed axosomatic synapses with CCK-negative axonal terminals. The CCK-positive dendrites and axons situated everywhere in the PVN. Some of them as postsynaptic elements formed axodendritic and axoaxonic synapses with CCK-negative structures. Some CCK-positive axonal endings surrounded the capillaries. Other CCK axonal terminals as presynaptic elements formed axosomatic, axondendritic and axo-axonic synapses with CCK-negative structures, respectively. In addition, we have first found that the CCK-positive dendrites penetrated ependyma and contacted directly with the cerebrospinal fluid in third ventricle, the CCK-positive axons traveled in the cavity of third ventricle near the ependyma. The above mentioned results suggested: (1) the soma, dendrite and axon of the CCK-containing neurons and CCK-negetive neurons in the PVN might form local neuronal circuit; (2) the neuron vessel circuit might be established between CCK-containing neurons and the blood vessels in the PVN; (3) the CSFcontacting neurons in the PVN may participate in forming brain-cerebrospinal fluid neurohumoral circuit and regulate functional activity of distal target area through the CSF pathway.

The distributions of neurotensin(NT) and substance P(SP) in the arcuate nucleus of rat hypothalamus have been studied by means of double labeled pre-embedding immunoelectron microscopic technique. It was observed that there were SP- and NT-containing dendrites, perikarya and axons in the arcuate nucleus. SP- and NT-containing dendrites and axons received asymmetric afferent synapses from immunonegative axons. SP-positive axonal terminals established symmetric axo-somatic and axo-dendritic synapses with immunonegative perikarya and dendrites as well as symmetric axo-somatic synapses with NT-positive perikarya. The results of this study directly indicate for the first time that the NT-ergic neurons in rat arcuate nucleus receive innervation from SP-ergic neurons, and provided an ultrastructural evidence for the synaptic regulation of the neuroendocrine of the hypothalamus.

The distribution of neurotensin (NT) and neuropeptide Y (NPY) in the rat hypothalamic arcuate nucleus has been studied ultrastructurally by means of double labeling preembedding immunoelectron microscopic PAP technique. First, the NPY immunoreaction was demonstrated by chromogen DAB, and second, the NT immunoreaction was demonstrated by ammonium molybdate-TMB method. After being stabilized by DAB-cobalt chloride, the vibratome sections were processed for electron microscopic study. The results showed that in the arcuate nucleus the NPY immunoreactive products appeared as high electron-dense granular or flocculent materials deposited diffusely in the organelles and matrix of perikaryon, around the dendritic microtubules and axonic small clear vesicles. Whereas the NT immunoreactive products were dense needle- or mass-like deposits distributed dispersively in the perikaryon, dendrites and axon terminals. They can easily be distinguished although being intermingled together. The NPY-containing dendrites and axons formed synaptic connections with immuno-negative axon terminals, NT-containing somata and dendrites formed also synaptic conections with negative axon terminals. In addition, NPY-positive axon terminals formed symmetrical axodendritic synapses with NT-positive dendrites. The present results provided another new ultrastructural evidence for the peptidergic synaptic regulation of NT neurons in hypothalamus.

Purpose　The aim is to isolate and purify Protein C(PC) and Protein S(PS) from no-albumin human plasma by rivanol precipitation. Methods　The isolated and purified steps included adsorption onto and elution from barium, salting-out, ion-exchange chromatography, affinity chromatography,preparative isoelectric focusing and so on. Results　The molecular weights of the obtained PC and PS were (61±0.9)kD and (83±0.8)kD, respectively, the isoelectric point, 4.70±0.03 and 5.20±0.03,and the yield, 28.3% and 12.6%. The purified PC and PS were shown to be highly homogeneous by capillary zone-electrophoresis(CZE), and rich in Glu, Leu and Gly or Asp, Glu and Leu respectively. Conclusion　 The methods could be used for large-scale isolation and purification of PC and PS from no-albumin human plasma.

BACKGROUND: The metal implant internal fixation exhibits good effect for femoral neck fractures of young patients, but few studies report the metal implant treatment for femoral neck fracture in the aged patients. OBJECTIVE: To compare different implant treatments for osteoporotic hip fractures in senile patients. METHODS: A total of 237 aged patients with osteoporotic hip fracture, aged from 60 to 96 years old, were recruited from the hospital between January 1998 and December 2008. There were 32 cases treated by cannulated screw internal fixation, 23 cases by anatomical plate internal fixation, 41 cases by dynamic hip screw internal fixation, 111 cases by bipolar femoral head replacement, and 30 cases by total hip replacement. Comparison of the incidence of complications and the recovery of hip function were performed in each group. RESULTS AND CONCLUSION: Cannulated screws, anatomical plates, dynamic hip screw internal fixation showed a higher complication rate significantly than bipolar femoral head replacement and total hip arthroplasty (P < 0.01); the good and excellent effect rate in cannulated screw, femoral end plate, and dynamic hip screw fixation was significantly lower than that in bipolar femoral head replacement and total hip arthroplasty (P < 0.001, P < 0.01). The results suggested that artificial joint replacement (bipolar femoral head replacement or total hip replacement) is the optimal choice for aged osteoporotic femoral neck fracture, cannulated screw fixation is suitable for Garden I, dynamic hip screw fixation and proximal femoral anatomical plate fixation fit for intertrochanteric fractures of Jensen-Evans I-Ⅱ; Jensen-Evans Ⅱ-Ⅲ osteoporotic intertrochanteric fractures joint replacement is the ideal choice.

Objective An ELISA-based assay for detecting alloantibodies against FⅧ was established to estimate the incidence of alloantibodies against FⅧ in treated patients with hemophilia A. Methods One hundred and forty patients with hemophilia A and 80 healthy controls were enrolled. Among hemophilia A patients, 84, 34 and 22 patients were in severe, moderate and mild conditions respectively. All patients were treated with plasma-derived FⅧ concentrates before. The titer wells were coated with MoAb against FⅧ which was developed in our laboratory. Then human recombinant FⅧ concentrates were applied. After incubation in room temperature for 2 hours, diluted plasma samples and HRP-conjugated goat anti-human IgG were added successively. Finally Absorbance (A490) were measured and recorded. Inhibitor activity against FⅧ for all plasma samples was measured by a modified Nijmegen assay simultaneously. Results The results showed that alloantibodies against FⅧ were found in 40.0% (56/140) patients by ELISA. And the alloantibody incidences in the severe and non-severe patients were 47.6% (40/84) and 28.5% (16/56)respectively. There was statistical significance between these two categories (x2 = 5.079, P ＜ 0.05 ). The FⅧ inhibitor activity was detected in 24.3% (34/140) patients by modified Nijmegen assay. The inhibitor incidences in the severe and non-severe patients were 33.3% (28/84) and 10.7% (6/56) respectively.There was statistical significance (x2 = 9.349, P ＜ 0.05). Twenty-five patients were positive for FⅧ alloantibodies by ELISA but had no FⅧ inhibitor activity by the modified Nijmegen assay. The positive rates of FⅧ alloantibodies and inhibitor activity were 40.0% (56/140) and 24.3% (34/140) respectively,which had significant difference (x2 = 15.75, P ＜ 0.01 ) and strong positive correlation ( rn = 0.59, P ＜0.01 ). Meanwhile the results deduced from these two tests shared a high consistency rate ( Kappa = 0.55,P ＜0.01 ). Conclusions The detection rate for alloantibodies against F Ⅷ is enhanced by our newlydeveloped ELISA. Our results suggest that the occurrence of the alloantibodies against F Ⅷ in Chinese hemophilia A patients is not rare and the alloantibody incidence is preponderant in the patients with severe hemophilia A compared with non-severe hemophilia A patients.