Skin-visceral divergent projections of cholecystokinin(CCK)-containing dorsal root ganglion neurons were studied by combined technique of fluorescent double-labelling and immunohistochemistry. Fast Blue(FB) and Nuclear Yellow (NY) were injected into the coeliac ganglion and the cutaneous branches of left 9th-11th intercostal nerves, respectively. Three kinds of neurons labelled with fluorescein were observed in T_(9-11) dorsal root ganglia under Nikon fluorescence microscope with 365 nm excitation light: FB-labelled neurons with blue-fluorescing cytoplasm; NY-labelled neurons with yellow-fluorescing nucleus and double-labelled neurons with blue cytoplasm and yellow nucleus. The double-labelled neurons were found to be 2.8％ of total labelled neurons.The sections containing fluorescein labelled neurons were then stained by CCKimmunohistochemical procedure. Four kinds of neurons could be identified: NY-neurons, with CCK-immunoreactivity (NY+CCK); FB-neurons with CCK-immunoreactivity(FB + CCK);NY + FB neurons with CCK-immunoreactivity(NY + FB + CCK); Single CCK-positive neurons. NY + FB + CCK triple-labelled neurons accounted for approximately 11.5％ of NY + FB double-labelled neurons,and 0.4％ of all CCK-positive neurons.The findings clearly indicated that the peripheral processes of some sensory dorsal root ganglion neurons project divergently to both skin and visceral structure, and contain CCK. The present results suggest that the peripheral dichotomization of the dorsal root ganglion nearons might converge sensory inputs from both skin and visceral fields, and thus not only provide one of the structural basis for the referred pain but also reveal that CCK might play a mediation role in the skin-visceral reflection and referred pain.

Objective To investigate the clinical outcomes of proximal femoral nail antirotation (PFNA) Ⅱ in the treatment of femoral intertrochanteric fractures.Methods From January 2011 to June 2016,240 patients with intertrochanteric fracture were treated at our department.They were 132 males and 108 females,aged from 30 to 106 years (average,71.4 years).Of them,168 cases were older than 70 years.According to AO classification,21 patients were of type 31-A1.1,15 of type 31-A1.2,27 of type 31-A1.3,39 of type 3 1-A2.1,51 of type 3 1-A2.2,32 of type 3 1-A2.3,12 of type 3 1-A3.1,23 of type 3 1-A3.2 and 20 of type 31-A3.3.The time from injury to surgery in this series ranged from 38 to 140 hours,averaging 52 hours.All the patients were treated by fixation with PFNA Ⅱ.Results The average operating time was 45 min,average operation incision 5 cm,and average blood loss during operation 85 mL.Primary wound healing was achieved in 239 patients,and one patient developed postoperative incision infection which was cured after implants were taken out and debridement was carried out 8 weeks after operation.This group obtained an average follow-up of 31.5 months (from 6 to 51 months).Primary fracture healing was achieved in 235 patients after 6 to 23 weeks (average,12.6 weeks);fracture healing was delayed in 5 patients after 36 to 96 weeks (average,56 weeks).There was no in-hospital mortality,and the survival rate 6 months after operation was 98.8％ (237/240).One year after operation,132 patients obtained an average Harris hip score of 85.2 (from 72 to 94).No implant-related complications or implant failure happened.Conclusions Internal fixation with PFNA Ⅱ is an effective treatment for different types of femoral intertrochanteric fractures,because it is simple and less invasive,and leads to limited complications,especially for senile patients.

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.

Origin and ultrastructural characteristics of serotonergic fibers of the spinal dorsal horn in the rat have been confirmed by means of a combined method of HRP and immunocytochemistry and immunoelectron microscopic observation. The results showed that serotonergic axonal terminals in the spinal dorsal horn come mainly from nucleus raphe magnus and the ventral part of the reticular formation of medulla oblongata. Serotonin immunoreactive positive structures of the spinal dorsal horn have been found in lamina Ⅰ (marginal zone) and lamina Ⅱ (substantia gelatinosa) as fine myelinated and unmylinated fibers. There were mainly axo-axonic synapses between the labeled and nonlabeled terminals. The labeled terminals were presynaptic or postsynaptic element. Axo-dendritic synapses were rarely found. The non-synaptic releasing figures have not been found. Based on the ultrastructural characteristics the authors suggest that in performing analgesia role the serotonergic system in the spinal dorsal horn might influence directly or indirectly the excitability of interneurons and inhibit directly the nerve impulses of primary afferents by means of synaptic connections instead of non-synaptic releasing manner.

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.

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.

Using the immunohistochemical and immunoelectron microscopic methods, the localization and pattern of vasoactive intestinal polypeptidergic nerves in the wall of cerebral vessels were investigated. The results revealed that the VIP-like immunoreactive (VIPLI) fibres were located in the adventitia or at the adventitia-media border. No synaptic contacts were identified among the nerve terminals or between nerve terminal and smooth muscle cell. VIPLI terminals directly apposed to the smooth muscle cells with a distance of 100 nm.The observations mentioned above indicate that there is peptidergic innervation in the wall of the cerebral vessels besides traditional adrenergic and cholinergic innervations. The regulatory function of VIP-containing nerves to the cerebral vessels may be performed by affecting the smooth muscle directly through the nonsynaptic release. In addition, present study identified and discussed the distribution and function of substance P(SP) in the cerebral vessels.

Objective To observe the antioxidative dosage-effect relationship of grape seed proanthocyanidin extract (GSPE). Methods The mice were randomly divided into seven groups:blank group, model group, 5 mg/kg GSPE group, 15 mg/kg GSPE group, 45 mg/kg GSPE group, 135 mg/kg GSPE group and 405 mg/kg GSPE group. The mice in blank group were dealt with saline solution by intraperitoneal injection, the others were dealt with D-galactose (120 mg/kg) by intraperitoneal injection for seven weeks to make oxidative damage model. Meanwhile, the mice were given corresponding dose of the drug. Subsequently the level of malondialdehyde (MDA) and activity of superoxide dismutase (SOD) in the serum were measured to observe the antioxidative dosage-effect relationship of GSPE. Results The 45, 135, 405 mg/kg GSPE group reduced the MDA level, and the 15, 45, 135, 405 mg/kg GSPE group increased the SOD activity. Conclusion GSPE has significant antioxidant activity on mice dealt with D-galactose above the dose of 15 mg/kg, suggesting that the clinical use of GSPE should guarantee a certain dose to play a good antioxidant effect.

Aim To investigate ALEX1 gene expres-sion in cervical cancer tissues and adjacent non-can-cerous tissues, and to explore the ALEX1 genetic influ-ence on cell proliferation,cycle and apoptosis of human cervical cancer cell line HeLa. Methods ALEX1 protein expression in cervical cancers and in non-can-cerous cervical tissues was evaluated using immunohis-tochemical method. A small interference RNA targeting ALEX1 gene was transfected into HeLa cells′, and the effect of ALEX1 interference on HeLa cells′ cycle and apoptosis was analysed by flow cytometry. The effect of ALEX1 interference on HeLa cells′ proliferation and sensitivity to resveratrol was analysed by CCK-8 assay. Results ALEX1 protein expression was significantly increased in cervical cancer tissues compared with non-cancerous tissues. HeLa cells′proliferation was inhibi-ted compared with control group and blank group. He-La cells′ sensitivity to resveratrol was enhanced com-pared with control group blank group. Conclution SiRNA silencing of ALEX1 gene could significantly in-hibit HeLa cells′ proliferation and enhance resveratrol ability of inhibiting HeLa cells′proliferation.