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This study explored the possibility that the components in melanoma cytoplasm induce murine BMSCs transformation and expression of Melan-A by morphologically observing the changes of BMSCs and immunocytochemically detecting Melan-A in the cells after culturing BMSCs in medium containing melanoma cytoplasm components (MCC). MCC of B16 melanoma cells was prepared and BMSCs were cultured and induced by adding the MCC into culture medium. The cells were morphologically observed and Melan-A was immunohistochemically detected to confirm BMSCs transformation. MCC-induced BMSCs underwent morphological changes. A number of melanin granules appeared in the cytoplasm of the cells and some were released into surrounding areas. Several cells that might come from one cell formed a cluster, and their granules, together with those secreted by other induced BMSCs, formed a so-called "sphere-formed structure". The induced BMSCs expressed Melan-A. We are led to conclude that there might be some factors in the cytoplasm of melanoma cells that might induce BMSCs transformation toward melanogenic cell, or even melanoma.
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This study investigated the ability of millimeter-wave (MMW) to promote the differentiation of bone marrow stromal cells (BMSCs) into cells with a neural phenotype. The BMSCs were primarily cultured. At passage 3, the cells were induced by beta-mercaptoethanol (BME) in combination with MMW or BME alone. The expressions of nucleostemin (NS) and neuron-specific enolase (NSE) were detected by immunofluorescent staining and Western blotting respectively to identify the differentiation. The untreated BMSCs predominately expressed NS. After induced by BME and MMW, the BMSCs exhibited a dramatic decrease in NS expression and increase in NSE expression. The differentiation rate of the cells treated with BME and MMW in combination was significantly higher than that of the cells treated with BME alone (P<0.05). It was concluded that MMW exposure enhanced the inducing effect of BME on the differentiation of BMSCs into cells with a neural phenotype.
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OBJECTIVE@#To investigate the expression of FasL and ICAM-1 on the HEVs in the hypertrophic tonsils and its clinical significance.@*METHOD@#Under the light microscopy, HE and immunohistochemistry methods were used to show the morphology of HEVs, the number of T, B lymphocytes, the expression intensity of FasL and ICAM-1 in 17 patients with the idiopathic tonsillar hypertrophy (ITH) and in 20 patients with the chronic hypertrophic tonsillitis (CHT).@*RESULT@#The expression intensity of ICAM-1 on the tonsillar HEVs in ITH group was significantly lower than that in CTH group (P < 0.05), but the intensity of FasL in ITH group was significantly higher than that in CHT group (P < 0.05).@*CONCLUSION@#FasL and ICAM-1 expression in the tonsillar HEVs contribute to the CHT and ITH. The former mainly makes IHT developed, and the latter promotes the process of the inflammation.
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Male , B-Lymphocytes , Allergy and Immunology , Endothelium , Fas Ligand Protein , Metabolism , Hypertrophy , Intercellular Adhesion Molecule-1 , Metabolism , Palatine Tonsil , Metabolism , Pathology , T-Lymphocytes , Allergy and Immunology , Tonsillitis , Metabolism , Venules , MetabolismABSTRACT
To examine the aging-related changes of microglia and astrocytes in hypothalamus of rats after intraperitoneal injection of hypertonic saline in rats, old- and young-aged rats were injected with hypertonic saline solution into peritoneal cavity. Lectin histochemical techniques using Ricinus communis agglutinin-1 (RCA-1) and immunocytochemical method employing antibody against glial fibrillary acidic protein (GFAP) were used to demonstrate microglia and astrocytes in the hypothalamus of the rats, and the positively-stained cells were analyzed by computer-assisted image analysis system. Our results showed that the numbers of microglia and astrocytes were significantly increased in the hypothalamus of old-aged rats. After intraperitoneal injection of hypertonic saline, the number of microglia was significantly decreased in the hypothalamus of both young- and old-aged groups. After introperitoneal injection of hypertonic saline, the number of GFAP positive cells was significantly increased in the hypothalamus of young rats, but the number of GFAP positive cells did not show significant change in the hypothalamus of old rats. It is concluded that in the hypothalamus of old-aged rats, the increase of microglia may be related with the aging or degeneration of neurons, and the increase of astrocytes may provide more nourishment required by the aged neurons. The microglia and astrocytes in the hypothalamus of the two group rats may be affected by hypertonic saline, and the response of these cells to the stimuli is characterized by some aging-related changes.
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BACKGROUND: Microglia is the immune effect cell in central nervous system, if it is activated, it will release neuron poisoning factors and inflammatory factors, and bring fatal injury.OBJECTIVE: To investigated the effect of focal cerebral reperfusion on activation of microglia and the effect of the regulation of electroacupuncturing Shuigou and Baihui.DESIGN: A randomized and controlled study.SETTING: The department of Physioltherapy, the 458 Hospital of Chinese PLA; Dongfang Hospital Affiliated to Shanghai Tongji University; The department of Histology-embryology, Tongji Medical College of Huazhong University of Science and Technology.MATERIALS: This experiment was processed at Histology-embryology Center of Tongji Medical College of Huazhong University of Science and Technology in March 2002. Total 80 Wistar rats were randomly divided into 8 groups with 10 in each group.METHODS: ① Normal control group: The rats were not given any treatment and killed next day. ② Sham operation group: Only artery was separated, no suture inserted, killed next day. ③ 6, 12 and 24-hour reperfusion groups: The rats' right middle cerebral artery (MCA) ischemia-reperfusion model was made by embolus suture technique, then the rat was killed after 30 minutes' occlusion at MCA and reperfusing respectively 6, 12, 24hours. ④ 6-hour reperfusion + electroacupuncture group: Electroacupuncture was carried out immediately after the model was made, then the rat was killedafter 30 minutes' occlusion at MCA and reperfusion 6 hours (The frequency was 4 Hz-16 Hz. Stimulus intensity increased 1 V per 10minutes and the final intensity was 3V. The stimulus was lasted 30 minutes). ⑤ 12-hour reperfusion + electroacupuncture B group: Electroacupuncture was carried out immediately after the model was made and 8 hours later, then the rat was killed after 30 minutes' occlusion at MCA and reperfusion 12 hours. ⑥ 24-hour reperfusion + electroacupuncture group: Electroacupuncture was carried out immediately after the model was made and 8, 16 hours later, then the rat was killed after 30 minutes' occlusion at MCA and reperfusion 24 hours.MAIN OUTCOME MEASURES: To calculate numbers of microglia and to observe its morphological changes.RESULTS: Data of totally 67 rats was entered the results analysis. No microglia was seen in the normal and sham operation groups. Large quantity of microglia was activated at the border of ischemic area in 6, 12 and 24reperfusion group, their quantity was largely increased, reaching apex at reperfusion 12 hours [(35.38±1.77), (54.25±1.67), (49.29±2.21)/200sights]. The quantity in every 6, 12 and 24 reperfusion + electroacupuncture group was less than that in model group [(32.11±2.80), (50.88±2.64),(45.45±3.95)/200 sights, P ＜ 0.05].CONCLUSION: The microglia in brain is activated after the cerebral ischemia-reperfusion and induces toxic effect on neurons. The electroacupuncture can decrease activation of microglia so as to protect the neurons.
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To examine the aging-related changes of microglia and astrocytes in hypothalamus of rats after intraperitoneal injection of hypertonic saline in rats, old- and young-aged rats were injected with hypertonic saline solution into peritoneal cavity. Lectin histochemical techniques using Ricinus communis agglutinin-1 (RCA-1) and immunocytochemical method employing antibody against glial fibrillary acidic protein (GFAP) were used to demonstrate microglia and astrocytes in the hypothalamus of the rats, and the positively-stained cells were analyzed by computer-assisted image analysis system. Our results showed that the numbers of microglia and astrocytes were significantly increased in the hypothalamus of old-aged rats. After intraperitoneal injection of hypertonic saline,the number of microglia was significantly decreased in the hypothalamus of both young- and oldaged groups. After introperitoneal injection of hypertonic saline, the number of GFAP positive cells was significantly increased in the hypothalamus of young rats, but the number of GFAP positive cells did not show significant change in the hypothalamus of old rats. It is concluded that in the hypothalamus of old-aged rats, the increase of microglia may be related with the aging or degeneration of neurons, and the increase of astrocytes may provide more nourishment required by the aged neurons. The microglia and astrocytes in the hypothalamus of the two group rats may be affected by hypertonic saline, and the response of these cells to the stimuli is characterized by some aging-related changes.
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This experiment aimed to investigate the effect of adrenergic system in the subnucleus commissuriu of nucleus solitrius tractus (CNTS) on renal nerve discharges. Norepinephrine (NE) was microinjected into the CNTS of rabbits and mean arterial blood pressure (MAP) and renal nerve discharges (FRND) were synchronously recorded. The results indicated that (1) microinjection of norepinephine into the CNTS of rabbit could significantly attenuate the frequency of renal nerve discharge, and at the same time decrease markedly the mean arterial pressure. (2) Microinjection of 0.3 nmol yohimbin into CNTS had no significant influence on FRND and MAP, but could attenuate and even reverse the effects of NE on FRND and MAP. These results suggest that microinjection of NE into CNTS may activate the alpha-adrenorecptor located in CNTS and secondarily produce a depressor effect by attenuating the activity of periphenal sympathetic nervous system.
Subject(s)
Blood Pressure/drug effects , Depression, Chemical , Kidney/innervation , Microinjections , Norepinephrine/pharmacology , Solitary Nucleus/physiology , Sympathetic Nervous System/drug effects , Sympathetic Nervous System/physiopathology , Vasomotor System/physiopathologyABSTRACT
Summary: This experiment aimed to investigate the effect of adrenergic system in the subnucleus commissuriu of nucleus solitrius tractus (CNTS) on renal nerve discharges. Norepinephrine (NE) was microinjected into the CNTS of rabbits and mean arterial blood pressure (MAP) and renal nerve discharges (FRND) were synchronously recorded. The results indicated that (1) microinjection of norepinephine into the CNTS of rabbit could significantly attenuate the frequency of renal nerve discharge, and at the same time decrease markedly the mean arterial pressure. (2) Microinjection of 0.3 nmol yohimbin into CNTS had no significant influence on FRND and MAP, but could attenuate and even reverse the effects of NE on FRND and MAP. These results suggest that microinjection of NE into CNTS may activate the alpha-adrenorecptor located in CNTS and secondarily produce a depressor effect by attenuating the activity of peripheral sympathetic nervous system.
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<p><b>OBJECTIVE</b>To examine the expression of the tau protein and microtubule-associated proteins in the testis interstitium of aged and young rats.</p><p><b>METHODS</b>Sprague-Dawley male rats were divided into a young group(6 months, n = 10) and an aged group(28 months, n = 10). The two steps immunohistochemistry method with the antibody against tau protein and MAP alpha was performed on the testis tissues of the rats.</p><p><b>RESULTS</b>The results showed that the immunoreactive cells of tau protein of the testis interstilial of the aged rats obviously increased(P < 0.001) than those of the young, while the immunoreactive cells of the microtubule-associated proteins decreased(P < 0.01) in the aged.</p><p><b>CONCLUSION</b>The changes in the expression of the tau protein and microtubule-associated proteins may be related to the aging process of the testis.</p>
Subject(s)
Animals , Male , Rats , Aging , Metabolism , Immunohistochemistry , Microtubule-Associated Proteins , Rats, Sprague-Dawley , Testis , Chemistry , tau ProteinsABSTRACT
The distribution and function of transforming growth factor-beta (TGF-beta) in the region of bone defect repaired by collagen/nano-beta-tricalcium phosphate composite artificial bone (Co/N-TCP) and the ability of Co/N-TCP recruiting osteoblasts to precipitate the repair of bone defect were investigated. Twenty-four domestic rabbits were operated on bilateral cranial bone to create an experimental bone defect of 8.0 mm in diameter through the whole bone. On the left, Co/N-TCP was implanted as experimental group, but on the right, Co/TCP was implanted as control group. At 2nd, 4th, 8th, 12th week after operation, all animals were sacrificed and the implanted materials with surrounding bone were taken out. Immunohistochemical staining was performed for TGF-beta assay by avidin-biotin complex method (SABC). Simultaneously, TGF-beta was quantitatively analyzed by HPIAS-1000 imaging analysis system. The immunohistochemical staining for TGF-beta revealed that osteoblasts and immature osteocytes highly expressed TGF-beta. Diffused TGF-beta positive staining particles appeared in the mesenchymal and fibrous-tissue. There was no significant difference in the TGF-beta positive staining between two groups in the medial region to original osseous beds at different time points (P > 0.05). However, in distal original osseous bed of the defected region, the positive expression of TGF-beta in the Co/N-TCP group was significantly stronger than in the control group (P < 0.05 or 0.01). The Co/N-TCP has good bioactivities and ability of stimulating and conducting TGF-beta to aggregate and precipitate the healing of bone defect.
Subject(s)
Animals , Rabbits , Biocompatible Materials , Therapeutic Uses , Calcium Phosphates , Ceramics , Collagen , Fracture Healing , Implants, Experimental , Nanotechnology , Osseointegration , Osteogenesis , Physiology , Skull Fractures , Metabolism , General Surgery , Transforming Growth Factor beta , MetabolismABSTRACT
In situ hybridization and immunocytochemical techniques were employed to examine the expression of matrix metalloproteinases-1 (MMP-1) and to identify the pattern of its distribution in rat pancreas. The results indicated that the signal of MMP-1 mRNA and MMP-1 positive immunoreaction were detected in some fiberoblasts around interlobular ducts and exocrine cell in margin acinus of some lobules, but the signal of MMP-1 mRNA and MMP-1 positive immunoreaction could not be detected in most of other acinus and islets of pancreas. It is concluded that the expression of MMP-1 in above cells of rat might play an important role in acinar proliferation and differentiation of rat pancreatic tissues.
Subject(s)
Animals , Male , Rats , Cell Division , Immunohistochemistry , In Situ Hybridization , Matrix Metalloproteinase 1 , Genetics , Pancreas , RNA, Messenger , Genetics , Rats, Sprague-DawleyABSTRACT
Objective:To investigate the difference in expression of FasL and ICAM-1 in the tonsillar HEV associated with IgA nephropathy (IgAN) and recurrent tonsillitis (RT).Methods:Human tonsils were obtained from the patients with RT (n=18) or IgAN (n=12).HE staining was used to show the morophometry of the tonsillar HEV.CD20+,CD3+ lymphocytes and distribution of FasL and ICAM-1 in the HEV were observed with immunohistochemical stained tissue section.The quantitatively experimental data from pathological image analysis were compared between IgAN and RT group.Results:(1)The expressing intensity of ICAM-1 was (0.435 5?0.002) in IgAN group and (0.239 5?0.010 4) in RT group,respectively (P0.05).Conclusion:A significant increase in the number of tonsillar HEV and increased lymphocytes in HEVs may indicate the abnormalities in peripheral blood in the patients with IgAN,thus leading to an overactivation of the tonsillar HEVs that induce Fas-dependent apoptosis in such patients.A stronger biological effective adhesion molecule promotes the transendothelial migration of lymphocytes across HEVs of palatine tonsil,which appears to be connected with the development of IgAN.
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Objective To investigate the estrogen receptor expression character on neurons and glial cells of rat at different ages. Methods Brain sections from normal and injured rats of different ages were chosen to observe the ER immunohistochemistry reaction in cerebrum. Results To compare with normal young rat group,the neurons in hippocampus dentate gyrus of normal aged rat group expressed much less estrogen receptors.There were many ER-positive glial cells appearing in hippocampus of brain injured was rat group,and the positive strain of young injured was much stronger than that of aged rat group.Conclusion The decrease of ER on neurons in dentate gyrus makes the aged more apt to neurodegeneration.The young rats may better utilize the neuroprotection of estrogen against injuries by increasing the amount of ER on glial cells.