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Objective To observe the relationships among the direct projection neurons from the lumbar spinal cord to the lateral parabrachial nucleus(LPB),calbindin D-28K(CB)-like immunoreactive (-LI) neurons,peripheral noxious information transmission as well as substance P receptor(SPR)-LI neurons. Methods Triple-labeled techniques were used by tetramethyl rhodamine(TMR) retrograde tracing combined with immunofluorescence histochemistry for CB-,SPR- or Fos protein.The stained sections were observed under a confocal laser-scanning microscope. Results 1.After injecting TMR to the unilateral LPB,a number of TMR retrogradely labeled neurons were mainly distributed in the lamina Ⅰ,lateral spinal nucleus(LSN),and regions around the central canal of the spinal cord(lamina X) of the ipsilateral spinal cord;2.CB-LI neurons were mainly found in the laminae Ⅰ and Ⅱ of the lumbar spinal cord,especially in the lamina Ⅱ in the dense distribution; 3.SPR-LI neurons were also mainly seen in the lamina Ⅰ,LSN and lamina X of the spinal cord.A few of the SPR-LI neurons were also distributed in the lamina Ⅱ;4.Fos-positive neurons were detected in the laminae Ⅰ and Ⅱ,lateral aspect of the lamina Ⅴ to Ⅶ of the lumbar spinal cord by injecting 5% formalin into the ipsilateral hindpaw;5.Triple-labeled neurons for TMR/CB/SPR or TMR/CB/Fos were mainly found in lamina Ⅰ,while a few of the triple-labeled neurons were also found in lamina Ⅱ of the dorsal horn.TMR/CB/SPR triple-labeled neurons accounted for 103%,98% and 146% of total population of TMR-,CB- or SPR-LI neurons in laminae Ⅰ and Ⅱ,respectively.On the other hand,TMR/CB/Fos triple-labeled neurons formed 118%,106% and 158% of the total population of TMR-,CB-LI or Fos-positive neurons in the laminae Ⅰ and Ⅱ,respectively.Conclusion\ The results indicated that in the laminae Ⅰ and Ⅱ of the lumbar spinal cord some neurons with CB-Like immunoreactivity transmitting the peripheral noxious information and projecting directly to the LPB might receive SPergic primary afferents.
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Objective To study the expression of substance P(SP) and substance P receptor(SPR) during the development of mice brains. Methods The expression of SP and SPR during the development of mice brains from embryonic day(E) 11 to postnatal day(P) 0 days was analyzed by immunohistochemical method. Results The expression of SP began at E11 and gradually increased until birth. The expression of SPR began at E11 and maintained stable expression until birth. SP mostly expressed at striatum and SPR mostly expressed at medullary raphe.Conclusion The expression of SP and SPR during the embryo brain stage may indicate that SP could be an important factor involved in the early organization and maturation of neuron.
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Objective To investigate whether there are changes of the mast cells (MCs) and substance P ( SP) -ergic nerve terminals under light and electron microscopy, and to elucidate their possible roles in the pathogenesis of irritable bowel syndrome (IBS). Methods In 19 normal controls, 22 diarrhea predominant IBS ( D-IBS) and 20 constipation predominant IBS (C-IBS) , biopsies were taken from the terminal ileum, ileocecal junction, ascending colon and sigmoid colon. The MCs and the substance P (SP)-ergic nerve terminals, substance P receptor ( SPR) were stained by histochemical and immunohisto-chemical method respectively, and color image analyzer investigated the results qualitatively and quantitatively. The structure relation between the MCs and SP-ergic terminals was studied through an ultra-microscopy using in situs embedding technique and a light microscopic study in serial sections. Results The number of MCs in the terminal ileum, ileocecal junction, and ascending colon was significantly elevated in IBS (P
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Aim To investigate the expression and regulation characteristics of the Substance P receptor(Neurokinin-1R, NK-1R) in human skin keratinocytes and fibroblasts. Methods HaCaT cells,a human keratinocyte cell line, and fibroblasts were cultured. The expression of NK-1R protein was examined by immunohistochemisury technique,and the mRNA level was detected by reverse transcriptase polymerase chain reaction (RT- PCR). The expression and regulation of NK-1R were measured by flow cytometry in HaCaT cells and fibroblasts treated with various stimuli and drugs. Results The expression of NK-1R existed in human keratinocyte and fibroblast, mainly located on cell membranes and cytoplasma. The NK-1R also expressed at HaCaT cells and fibroblasts transcription levels, and the mRNA levels in HaCaT cells were higher than that of fibroblasts. SP and IFN-? might upregulate the membrane expressions of NK-1R in both the two cells, while LPS might downregulate the expressions of NK-1R. Cetirizine and Spantide I can degrade the expressions of NK-1R in the two kinds of cells.Conclusions The human keratinocytes and fibroblasts can express NK- 1R at cell, protein and transcription levels, and the expression characteristics can be regulated by some inflammatory factors, it indicates the keratinocytes and fibroblasts were involved in the regulation of skin immune and NK-1R may play an important role in skin inflammation.