Objective To investigate the expression of thyrotrophin-releasing hormone receptor type-2(TRH-R2) in rat testis during postnatal development,and to discuss its function in the regulation of reproduction development. Methods Western blotting and the immunohistochemical ABC method were used to detect the expression and location of TRH-R2 in testes of 8,15,20,35,60 and 90-day-old rats. And the immunohistochemical results were analyzed with the image analysis system and statistical method. Results Western blot showed that TRH-R2 was expressed in 15- to 90-day-old rat testis. As for the immunohistochemical ABC method,we found that TRH-R2 firstly appeared in 8-day-old rat testes. In all the following stages we also detected the immunochemical positive cells in testes. TRH-R2 was distributed in the interstitial cells between seminiferous tubules and positive substance was expressed in the cell membrane and cytoplasm of Leydig cells.The results of image analyses quantitation revealed that the intensity of TRH-R2 increased progressively with the postnatal development of rats.The peak was in 35-day-old.Then at the following stages the expression of TRH-R2 maintained stable.Conclusion TRH-R2 is expressed in rat testis during its postnatal development and locates in the Leydig cells,and the TRH-R2 shows development-dependent expression in testicular tissue.

Pancreastatin, a novel peptide isolated from porcine pancreas, is capable of inhibiting the insulin release and the exocrine secretion of the pancreas. In this study, the localization and distribution of pancreastatin in guinea pig, pig and human pancreas were investigated with ABC immunostaining method combined with the glucose oxidase-DAB-nickle developing technique on paraffin sections. The results showed that in human pancreas pancreastatin-like immunoreactive(PLI)cells were mainly dist- ributed in the periphery of the islets. While in guinea pig and pig pancreas, the majority of islets cells showed positive pancreastatin immunostaining. The coexistence of pancreastatin and insulin in the pancreatic islets of guinea pig and pig were also proved with immunostaining of adjacent thin sections. PLI-cells were also found in the epithelium of the ducts and acini of all investigated pancreas. However, in the exocrine portion of human and pig pancreas, PLI-cells were present in amounts smaller than those found in guinea pig pancreas. The significance of the distribution pattern of pancreastatin was discussed.

The distribution of neuropeptide Yimmunoreactive (NPY-IR) fibers in the spleen of rat was investigated by using the immunohistochemical ABC method combine with glucose oxidase-DAB-nikel enhancement technique. The results demonstrated that the rat spleen is richly innervated by NPY-IR fibers. The NPY immunoreactive nerve fibers are mainly distributed along the branches of the arteries, especially surrounding the central arterioles. They are also observed in the connective tissue capsule, the lymphoid tissue of the white pulp, red pulp and marginal zone and around the splenic sinusoid. The positive fibers are closely related to the blood vessels and lymphocytes in the spleen, which suggested that these fibers may regulate the development and function of splenic lymphocytes. NPY may have a direct or an indirect effect on regulating blood circulation of the spleen and the lymphocytes.

In this study, the distribution of calcitonin gene-related peptide immunoreactive (CGRP-IR) nerve fibers in the spleen of the rat was investigated by using the immunohistochemical ABC method combined with glucose oxidase-DAB-nickel enhancement technique. The results showed that the rat spleen was richly innervated by CGRP-IR fibers. The nerve fibers containing CGRP were mainly distributed in the red pulp, especially surrounding the splenic sinusoid and small blood vessels. They were also observed in the lymphoid tissue of marginal zone, and occasionally in the periarterial lymphatic sheath (PALS). The positive fibers distributed in the spleen, which suggested that the fibers might modulate the development and function of splenic lymphocytes.

Objective To study the localization of Smad 2 and Smad 4 proteins, which are intracellular signaling molecules of transforming growth factor ? family in adult rat testis. Method Immunohistochemical ABC method with glucose oxidase DAB nickel enhancement technique was used in the present study. Results Smad 2 immunoreactivity was mainly located in the spermatogonia, spermatocytes, spermatids, Sertoli cells in the seminiferous tubule and Leydig cells in the interstitial tissue. The reactive substance distributes in cytoplasm with negative nuclei. While Smad 4 is mainly expressed in cytoplasm of Leydig cells, and Sertoli cells is weak stained. Conclusion Our findings of Smad 2 expression in spermatogenic cells and Smad 4 expression in Leydig cells provide direct evidence for the molecular mechanism of TGF ? action during spermatogenesis.

ObjectiveTo investigate the expression of thyrotropin-releasing hormone receptor (TRH-R) type-1and type-2 in ethane dimethanesulphonate (EDS)-treated rat testis, and to discuss the significance of its expression in Leydig cells.Methods To make the injured testis Leydig cells rat model with EDS treatment. Western blotting, immunohistochemical ABC and immunofluorescence double labeling methods were used to detect the expression and location of TRH-R1 and R2 in the testicular tissues of EDS-treated-day 2,day 7,day 14,day 21 and day 28 rat mode, respectively. Results Western blotting results showed that the positive immunochemical staining was not found in the testicular tissues of the EDS-treated day 2 to day 14, on the other hand,they were found in EDS-treated-21 day and EDS-treated-28 day. Immunohistochemistry demonstrated that TRH-R1 and R2 expressed in the spindle-shaped cells reappeared around seminiferous tubules of post-EDS 21 days and 28 days groups. Immunofluorescence double labeling confirmed that these TRH-R1 and R2 positively stained cells were newly regenerated progenitor Leydig cells.Conclusion TRH-R1 and R2 are involved in the regeneration of Leydig cells in EDS-treated rat testis, and they may exert functions in the proliferation and differentiation of adult type Leydig cells.

0.05).CONCLUSION:Nedaplatin combined with docetaxel is effective and safe for advanced NSCLC with high short-term efficacy and mild toxic side reaction in digestive tract and kidney.but the long-term efficacy of it require further study.

Objective To investigate the expression of histamine in the cardiac sympathetic fibers from the guinea pig superior cervical ganglion and its coexistence with norepinephrine so as to provide morphological evidence for histamine as a cardiac sympathetic neurotransmitter.Methods Biotinylated dextranamine(BDA) anterograde tracing and immunofluorescence histochemical staining for histamine/norepinephrine were applied.Results After injection of BDA into the superior cervical ganglion,BDA labeled sympathetic fibers in the left and right atria and ventricle were observed.Meanwhile,the tracing fibers proved histamine-like immunoreactive or both histamine and norepinephrine-like immunoreactive.Conclusion Histamine is expressed in the cardiac sympathetic fibers from the guinea pig superior cervical ganglion and coexisted with norepinephrine.

Objective To confirm the localization and function mechanisms of TGF?s and their receptors in mouse spermatogenesis,by observing the expression of TGF?s and their receptors in mouse testicular tissues. Methods Immunohistochemical staining combined with the micro-image analysis and immunofluorescence histochemical double-staining technique were used. Results Immunohistochemical staining display:Both TGF?1 and TGF?2 were expressed in acrosome of Ⅵ-Ⅷ stages round spermatids and elongated spermatids in seminiferous tubules,in cytoplasm of Leydig cell their expression was very weak;TGF?3 was only expressed in the cytoplasm of Leydig cells and there was no positive expression in seminiferous epithelium;TGF?-RⅠ was expressed in late spermacytes(Ⅷ stage) and early elongating spermatids(Ⅹ stage);TGF?-RⅡ was expressed from the Ⅺ stage elongating spermatids to elongated spermatids released;TGF?-RⅢ was expressed in acrosome of late round spermatids (Ⅶ-Ⅷ stages)and elongated spermatids.Immunofluorescence histochemical double-staining display:the earlist co-expression of TGF?1,TGF?2 and TGF?-R3 in seminiferous epithelium was at acrosome of Ⅵ-Ⅷ stages round spermatids.Conclusion TGF?s and their receptors are expressed with cell-and stage-specifity in mouse spermatogenesis,which suggests that they have an important role in spermatogenesis.

Objective To examine the localization of neurokinin B receptor (NK3)\|like immunoreactivity (\|LI) in the central nervous system of the mouse. Methods An immunohistochemcial staining method was used. Results NK3 receptor\|LI was localized in somatic and dendritic profiles in the most parts and in neuropil in a few regions of the mouse central nervous system. A large number of neurons with NK3\|LI was seen in the anterior olfactory nuclei, accumbens nucleus, septal area, ventral pallidum, pallidum, caudate putamen, nucleus of the stria terminalis, anterior hypothalamic area, tuber cincreum area, lateral hypothalamic area, perifornical nucleus, supraoptic nucleus, arcuate nucleus, mammillar nuclei, substatia nigra, ventral tegmental area, retrorubral area, superior and inferior colliculus, periaqueductal gray, nucleus of the solitary tract, and superficial layers of the medullary and spinal dorsal horns. The superfical layers of the cerebral cortex, piriform cortex, dorsal hippocampus, amygdal complex, reticular formation of the brainstem contained some neurons with NK3 receptor\|LI. In the ventral hippocampus, median and intralaminar nuclei of the thalamus and interpeduncular nuclei, NKR\|LI was localized in neuropil. Conclusion\ Neurons with NK3 receptor\|LI were widely distributed in the central nervous system. It may be involved in many physiological functions in the central nervous system of the mouse.\;[