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Objective To explore the double-labeling method of monitoring the GHRP regulatory function on [Ca2+]i and NO in cardiomyocytes of rats on real time under LSCM.Methods The reformed constant-flow Langendorff system and enzyme-dissociated was used to isolate cardiomyocytes.[Ca2+]i and NO in the cardiomyocytes of SD rats were double-labeled by their molecular probe Rhod-2/AM and DAF-FM/DA,respectively to monitor the regulatory function of GHRP on [Ca2+]i and NO on real time by LSCM.Results Ca2+ signal showed a red fluorescence and NO showed a green fluorescence while the overlapping of the two signals showed a yellow-green fluorescence by this system,and the similar effect presents in both double-labeled state and the single labeled one:GHRP induced a transient[Ca2+]i increase then followed by a plateau phase while there was not significant change in NO signal system after GHRP stimulation under the LSCM in the cardiomyocytes of rats.Conclusions After having established the double-labeling method we monitored the GHRP regulatory function on [Ca2+]i and NO on real time in cardiomyocytes of rats under LSCM causing the [Ca2+]i biphasic increase while no significant change in NO signal system.
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Objective To observe the expression of PECAM-1 in colorectal cancer in rat and to explore the mechanism of lymphatic metastasis of tumor. Methods Wistar rats were used to make the animal model of primary colorectal cancer,by using HE staining,the cancer type and progression of cancer were devided.By immunohistochemistry staining,the expression of PECAM-1 in tumor tissue,blood vessels and lymphatic vessels were observed. Results PECAM-1 expression in blood vessels and lymphatic vessels were observed in normal tissue,while its expression on tumor tissue and lymphatic vessels decreased according to the cancer progression.Conclusion PECAM-1 may be involved in the early process of tumor cells adhering to the endothelium;decreased expresson of PECAM-1 on lymphatics may be related with lymphangiogenesis and the opening of endothelial junction of lymphatic vessels.
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Objective To analyse the expression patterns of the vascular endothelial growth factor receptor 3(VEGFR-3) at the protein and mRNA level,and to detect the biological function of VEGFR-3 in the lymphangiogenesis of the embryos. Methods A total of 61 specimens of skin were investigated by immunohistochemical staining with a polyclonal antibody against VEGFR-3 in embryo 15-day-old(E15) and in embryo 21-day-old(E21).The expression of VEGFR-3 mRNA was studied in situ hybridization. Results The positive expression of VEGFR-3 can be seen in lymphatic vessels of the embryonic skin.The occurrences of VEGFR-3 protein in lymphatic vessels in E15 and in E21 were 38.71%(12/31) and 73.33%(22/30) respectively,the expression level of VEGFR-3 protein in E21 was significantly higher than that in E15(?~2=7.408,P
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Objective To study the ultrastructural features of initial lymphatics of gastric carcinoma,to discuss the metastasis mechanism of malignant cells by lpmphatics. Methods The postoperative samples of patients with gastric cancer were obtained in center,peripheral and normal region of cancer.Common Resin-embedded specimens were made.Lymphatics were located by hemithin sections under light microscope and observed by ultrathin sections under electron microscope. Results There were no lymphatics in carcinomatous centre region.Compared with normal region,there were more initial lymphatics in peripheral region of gastric carcinoma.The lumina were dilated and irregular and the endothelium were destroyed;The organelle of endothelial cells changed obviously.The distribution and morphology of the lymphatics did not changed evidently in the normal region.Conclusion These results suggest that amount,ultrastructure and morphology of lymphatics changed in peripheral region of carcinoma;Gastric cancer cells may invade lymphatics via destroyed parts of endothelium or interendothelial openings.
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The projection from the ventral posterior nucleus (VP) of the thalamus to the somatic sensory cortical area (SmI) in the rat was studied with the horseradish peroxidase (HRP) method.When injections were made into the anterior part of SmI (Brodmann 3), labeled cells were found in the ventral area of caudal VP. When injections were made into the posterior part of SmI (Brodmann 3,1), Iabeled cells were observed in the dorsal area of caudal VP. With injections located near the superior part of SmI (Brodmann 3), labeled cells were found in the lateral area of VP. When injections were made into the inferior part of SmI (Brodmann 2), labeled cells were seen in the medial area of VP.All labeled cells were observed in the injected side, indicating that the projection is uncrossed.Neurons in VP Projecting to Brodmann 2 is less than those projecting to Brodmann 3.Relationship between the quantity of labeled neurons and the amount of projection fibers was discussed.
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The projection from the thalamus and mesencephalon to the cingulate gyrus in the rat was studied with the horeseradish peroxidase method. Results were as follows:When injections were made into the anterior part of the cingulate gyrus, the labeled cells were found mainly in the thalamic nuclei (AM, MD, LD) and the midbrain nuclei (SNC, AVT, nuclei raphe dorsalis and centeralis superior). When injection was made into the posterior part of the cingulate gyrus, a considerable number of HRP-positive neurons were seen in the laterodorsal nucleus (LD), mediodorsal nucleus (MD) and anterior thalamic nuclei (AM, AV, AD).When the injection was made into one side of the cingulate gyri, all labeled cells were found in the ipsilateral side, indicating the projection is uncrossed.
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Objective To study the prelymphatic pathway in intestine. Methods Sudan black B was used to label fat which was fed to rats, then the pathway by which fat got into lacteals was observed under light and electron microscope. Results Under light microscope, we saw the blue\|greenish fat in epithelia, between epithelia, in lacteals and in the interstitial space around the lacteals. Under electron microscope, the pellets existed in epithelia, interstitial space and lacteals. Conclusion Fat might get into lacteals through the epithelia and the interstitial space which might be regarded as prelymphatic pathway. [
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30nm) accounted for 3.5% of these contacts. The mean maximum diameter of the vesicles was 49.4 nm, the volume density was 0.023 and profile density was 36. The distribution of the vesicles was as following: 29.1% opened onto or touched the abluminal membrane, 44,9% without visible connection to either surfaces, 26% opened onto or touched the luminal membrane. It was concluded that the mechanism of lymph formation in the stomach seems to be different from that of those organs such as heart, liver, kidney and thyroid. The present study suggested that the intercellular pathway may act as the primary route for lymph formation and macromolecule transportation, and the intracytoplasmic vesicular system may also play certain role in the transport of macromolecules.
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Objective To study the morphological changes of lymphatics in transplant tumor of mice. Methods H22 cells were inoculated under the armpit skin of KM mice. Tumor was obtained two weeks after inoculation. HE staining was used to observe the development of tumor. 5'-nase-alpase double staining was performed to observe blood vessels and lymphatics. Ultrastructures of lymphatics were observed with electron-microscopy. Results Tumor occurred in all the mice. Under microscope there was none of lymphatics in the center of the tumor, a few lymphatics and a lot of blood vessels could be seen in periphery of tumor. Under TEM damage of the organelles could be seen in the endothelial cell of lymphatics. Conclusion Neonatal lymphatics can be seen in transplant tumor caused by H22.
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By using 5'-nucleotidase-alkaline phosphatase (5'-Nase-AlPase) double staining method, the fine distribution of the lymph vessels in the stomach wall of the rabbit, guinea pig and rat has been observed. Under light microscope lymph capillaries showed strongly 5'-Nase positive reaction with brown or dark brown staining. However blood capillaries revealed significantly A1Pase activity with blue staining. The lymph capillaries were seen in all layers of the stomach wall in three species, but the lymph vessels were only shown in the submucosa, muscularis and serosa. In the mucosa lymph capillaries could be found in the deep layer of the lamina propria. In the submucosa there were lymph vessels and capillaries. In the muscularis lymph capillaries and vessels not only could be found between oblique and circular muscle or between circular and longitudinal muscle, but also among muscle fiber bundles of each layer. In the serosa there were larger lymph vessels situated near the muscularis. The observation on semithin and ultrathin sections also shown the similarity in the lymph vessels distribution. There was no difference in the distribution of lymph vessels in the stomach wall between rabbit, guinea pig and rat.