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2007, Volume 38 Issue 3
06 June 2007 Previous Issue    Next Issue
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MODIFIED RECOMBINANT HUMAN aFGF PROTECTS TYROSINE HYDROXYLASE NEURONS IN SUBSTANTIA NIGRA OF PARKINSON DISEASE RATS FROM LOSS 2007, 38 (3):  253-258.  doi: Abstract ( )   Objective To observe the changes of rotation behavior and tyrosine hydroxylase immunopositive neurons and investigate how Mrh-aFGF affects them in substantia nigra of Parkinson disease rats. Methods After building a rat model of Parkinson disease by injecting 6OHDA into substantia nigra and ventral tegmental area, we used Mrh-aFGF to intervene rats by lateral ventricle injection to observe how behavior of rats induced by apomorphine and tyrosine hydroxylase immunopositive neurons in substantia nigra of rats changes, then quantitatively analyzed the change of tyrosine hydroxylase immunopositive neurons. Results Rotation behavior was not found in control group, otherwise, actuation time was shorted, time length was prolonged, and average velocity of rotation was accelerated in Parkinson disease rats(P<0.01). Compared with PD group, rotation behavior of rats treated by using NS was not improved, however, rats treated with Mrh-aFGF, showed lengthened actuation time, shorted time length and velocity(P<001). With regard to immunopositive neurons of tyrosine hydroxylase, their numbers kept in a similar level in uninjured side of all groups, and no changes were found in bilateral substantia nigra of control group, however, the immunopositve neurons were decreased significantly in Parkinson disease, NS and MrhaFGF group (P<0.01). Regarding the injured side, tyrosine hydroxylase immunopositive neurons were gradually decreased in PD group(P<0.01), and similar changes were manifested in NS group. After treated with Mrh-aFGF, the structure of substantia nigra was significantly improved and the number of tyrosine hydroxylase neurons was increased, compared with Parkinson disease and NS group(P<0.01). Conclusion Mrh-aFGF could protect immunopositive neurons of tyrosine hydroxylase from loss and improve the rotation behavior of Parkinson disease rats Related Articles | Metrics

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POSTNATAL DEVELOPMENT OF S100B AND GFAP IN THE CENTRAL NERVOUS SYSTEM IN RATS 2007, 38 (3):  259-264.  doi: Abstract ( )   Objective To study the expressions of S100B and glial fibrillary acidic protein (GFAP) in the central nervous system of Sprague_Dawley rats during postnatal development. Methods Twenty_four male SD rats were divided equally into four groups according to different postnatal times:7_day group, 14_day group, 21_day group and adult group.Immunohistochemistry was used to investigate the expressions of S100B protein and GFAP in the brain and the spinal cord. Results The amount and density of S100B positive astrocytes decreased significantly in frontal cortex, hippocampus, striatum, substantia nigra and spinal cord during postnatal development. It seemed that the second through the third week after birth was a critical period for these changes. The amount and density of GFAP positive AST increased gradually in the brain, but it was the opposite in the spinal cord. Double_labeled immunofluorescence of S100B and GFAP in hippocampus CA1 area showed that S100B positive stains were evenly distributed in the pyramidal, polymorphic and molecular layers from the seventh till the twenty first day after birth, but apparently decreased in each layer especially in the molecular layer in adult hippocampus while the immunoreactivities of GFAP increased. The proportion of double labeled cells also increased with the aging and more of them were found in the pyramidal and polymorphic layers.Conclusion Different patterns of the expressions of S100B and GFAP exist during postnatal development. It may reflect the different roles of these proteins on the glial cell development. And it may also indicate that the expressions of S100B and GFAP are regulated by different mechanisms during the course of development, which reflects the differentiation of subpopulations of astrocytes during ontogenesis. Related Articles | Metrics

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THE EXPRESSION AND EFFECTS OF HEPCIDIN IN THE MOUSE BRAIN AND ITS MODULATING EFFECTS ON FERROPORTIN 1 AND DIVALENT METAL TRANSPORTER 2007, 38 (3):  265-270.  doi: Abstract ( )   Objective To investigate the distribution and role of hepcidin on ferroportin1 and divalent metal transporter 1(DMT1) expression in the mouse brain. Methods The expression of hepcidin in the mouse brain was detected by RT_PCR. The role of hepcidin on ferroportin1 and DMT1 expression was studied with cerebral ventricle injection and immunohistochemistry methods. Results There was hepcidin mRNA expression in the brain, and the level of hepcidin mRNA varied in different brain areas. Abundant expression of hepcidin existed in the choroid plexus.Conclusions The positive staining of DMT1, ferroportin1 was changed remarkably after the injection of hepcidin into cerebral ventricle of the brain; the change of DMT1 and ferroportin1 staining was regionally distinct. Related Articles | Metrics

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EFFECTS OF TRANSPLANTED OLFACTORY ENSHEATHINGCELLS AND L-NNA ON NEURONAL SURVIVAL AND AXONREGENERATION IN CLARKE’S NUCLEUS AFTER SPINAL CORD HEMISECTION OF RAT 2007, 38 (3):  271-277.  doi: Abstract ( )   Objective To investigate the effects of transplanted olfactory ensheathing cells (OECs)and L-NNA on neuronal survival and axon regeneration in Clarke’s nucleus (CN) after spinal cord hemisection of rats. Methods Twenty rats were divided into control group, L-NNA group, OECs group, and OECs+ L-NNA group after spinal cord hemisection. OECs were transplanted into the injured site of spinal cord and L-NNA was injected in OECs+L-NNA group of rats. Thirty days after operation, NADPH enzyme histochemistry, immunohistochemistry, neutral red staining, HE staining and fluorogold retrogradatic labeling were applied to determine the survival and axon regeneration of neurons at L1 CN in 4 groups of rats. Results 1. The number of neuronal survivals at L1 CN in OECs group and L-NNA group was higher than that in control group. The effect of promoting the survival of big, middle and small neurons at CN of spinal cord in OECs group and L-NNA group was shown by differential counts. The effect of promoting neuronal survival was the most significant in OECs+L-NNA group. 2. The regenerated axons were observed at the injured site of spinal cord and the neuron bodies labeled by fluorogold retrogradation were found at L1 CN in OECs group and OECs+L-NNA group. 3. Compared with those in control group, nitric oxide synthase (NOS) positive neurons were increased in OECs group, decreased in L-NNA group, and were no difference in OECs+L-NNA group.Conclusion Transplanted OECs or L-NNA may facilitate the survival of injured CN neurons of spinal cord. The combination of transplanted OECs and L-NNA can preferably promote the survival and axon regeneration of injured CN neurons Related Articles | Metrics

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EFFECTS OF INTRACEREBROVENTRICULAR ADMINISTRATION OF STREPTOZOTOCIN ON THE EXPRESSION OF PSD-95 AND Shank 1 IN RATS 2007, 38 (3):  278-281.  doi: Abstract ( )   Objective To investigate the effects of intracerebroventricular（ICV） administration of streptozotocin (STZ) on expression of PSD-95 and Shank1 in rat hippocampus. Methods A total of 30 rats were randomly divided into normal control group and model group. The rats in the model group received intracerebroventricular injection of STZ bilaterally with a dose of 3mg/kg and the injection was repeated on the third day. The rats in normal control group received the same treatment as the first group except for STZ injection. After 21 days, the brains of the rats were taken, frozen sections of brain were prepared, and then PSD-95 and Shank1 were stained with immunohistochemistry. Furthermore, the contents of PSD-95 and Shank1 were detected by Western blotting. Results After ICV injection of STZ, the expression of PSD-95 and Shank1 positive cells and the contents of PSD-95 and Shank1 in hippocampus both decreased. Conclusion ICV injection of STZ can lead to the decrease of the expression of PSD-95 and Shank1, indicating it may damage the neuron’s synaptic signal Related Articles | Metrics

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THE EFFECT OF CGRP ON THE EXPRESSION OF CREB mRNA IN RAT HIPPOCAMPUS AND PARIETAL CORTEX DURING FOCAL CEREBRAL ISCHEMIA AND REPERFUSION 2007, 38 (3):  282-285.  doi: Abstract ( )   Objective To investigate the effect of calcitonin gene related peptide (CGRP) on the expression of cyclic AMP response element binding protein (CREB) mRNA in rat hippocampus and parietal cortex during focal cerebral ischemia and reperfusion (I/R). Methods Focal cerebral ischemia/reperfusion model was induced by occluding of the right middle cerebral artery using the intraluminal suture method. Hybridization in situ experiment was used to detect the expression of CREB mRNA in the ipsilateral hippocampal CA1 region and parietal cortex during different reperfusion periods.The positive product of CREB mRNA was analyzed by image analysis system. Results There was a distinct expression of CREB mRNA in right hippocampal CA1 region and parietal cortex in sham group. The absorbency of CREB mRNA positive product reduced in I/R group as compared to sham group,while it increased in CGRP group than I/R group (P<0.05). Conclusion CGRP increases the expression of CREB mRNA in ischemic neurons of the hippocampus and parietal cortex during focal cerebral ischemia and reperfusion. CGRP’s protection of ischemic neuron may be conducted by activating the transcription and translation of CREB and then initiating a Related Articles | Metrics

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EFFECT OF HYPERBARIC OXYGEN ON THE EXPRESSION OF NEUROGLOBIN IN MOUSE HIPPOCAMPUS AFTER TRANSIENT FOREBRAIN ISCHEMIA 2007, 38 (3):  286-290.  doi: Abstract ( )   Objective To study the effect and significance of hyperbaric oxygen treatment on the expression of neuroglobin by observing the changing tendency of neuroglobin in mouse hippocampus after forebrain ischemiareperfusion. Methods Eighteen female C57BL/6N mice were divided into three groups randomly: the shamoperated control group, the 48hour ischemiareperfusion group (48h I/R) and the group treated with hyperbaric oxygen after 48hour ischemiareperfusion (48h HBO). HE staining and immunohistochemistry experiments were carried out on the brains of all the animals. Image analysis and statistical treatment were also performed for all the images. Results The neuroglobin expression level of 48h I/R group and the 48h HBO group were higher than that of the shamoperated group (P<0.01 and P<0.001, respectively); the optical density of the 48h HBO group was higher than that of the 48h I/R group(P<0.01).Conclusion Hyperbaric oxygen treatment had an obvious effect on ischemic brain injury, and increasing t Related Articles | Metrics

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REGULATION OF α-7 NEURONAL NICOTINIC ACETYLCHOLINE RECEPTORS BY NICOTINE IN MIDBRAIN OF RATS 2007, 38 (3):  291-295.  doi: Abstract ( )   Objective To observe the regulation of α-7 neuronal nicotinic acetylcholine receptors(α-7 nAChRs) by nicotine in the ventral tegmental area(VTA), the substantia nigra(SN) and the cortex of the rat. Methods Nicotine exposed animal models were established. The changes of α-7 nAChRs proteins and mRNA were observed by immunohistochemistry and in situ hybridization in VTA,SN and cortex, and the expression of α-7 nAChRs proteins by Western blotting in PC12 cells. Results The expression of α-7 nAChRs proteins in cortex and mRNA in VTA and SN were upregulated by nicotine. The expressions of α-7 nAChRs in PC12 cells were in proportion to and dependent on the time Related Articles | Metrics

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ES CELLDERIVED EPIDERMAL STEM CELLS FOR TREATMENT OFMICE FULL THICKNESS SKIN DEFECTS 2007, 38 (3):  296-299.  doi: Abstract ( )   Objective To provide a new way for treatment of full thickness skin defects by embryonic stem (ES) cellderived epidermallike stem cells. Methods Epidermal like stem cells, labeled by Hoechst 33342 and carried by a layer of biomembrane, were transplanted into the defective skin of mice. The differentiation tissue of donor cells was sampled each week. The sections were observed with HE staining, immunohistochemical and dilabeled immunofluorescence methods to test the expression of β1 integrin, CK15, CK19, CK10, CEA. Results The full thickness skin defects were healed in 2 weeks. The newborn skin was thicker than the normal skin. The basal layer cells proliferated.There were more bulky cellular poles towards dermis. The cells labeled by Hoechst 33342, located in the newborn epidermis and tubular or follicular structures in dermis, expressed β1 integrin and CK15 positive respectively in the first 3 weeks. There were sweat glandlike, sebaceous glandlike and hair folliclelike structures in the newborn dermis after 4 weeks. Basal cells of keratinized stratified squamous epithelium expressed CK19 and CK10 positive respectively and sweat glandlike structure expressed CEA positive. Conclusion ES cellderived epidermal stem cell Related Articles | Metrics

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ES CELLDERIVED EPIDERMAL STEM CELLS FOR TREATMENT OFMICE FULL THICKNESS SKIN DEFECTS 2007, 38 (3):  296-299.  doi: Abstract ( )   Objective To provide a new way for treatment of full thickness skin defects by embryonic stem (ES) cellderived epidermallike stem cells. Methods Epidermal like stem cells, labeled by Hoechst 33342 and carried by a layer of biomembrane, were transplanted into the defective skin of mice. The differentiation tissue of donor cells was sampled each week. The sections were observed with HE staining, immunohistochemical and dilabeled immunofluorescence methods to test the expression of β1 integrin, CK15, CK19, CK10, CEA. Results The full thickness skin defects were healed in 2 weeks. The newborn skin was thicker than the normal skin. The basal layer cells proliferated.There were more bulky cellular poles towards dermis. The cells labeled by Hoechst 33342, located in the newborn epidermis and tubular or follicular structures in dermis, expressed β1 integrin and CK15 positive respectively in the first 3 weeks. There were sweat glandlike, sebaceous glandlike and hair folliclelike structures in the newborn dermis after 4 weeks. Basal cells of keratinized stratified squamous epithelium expressed CK19 and CK10 positive respectively and sweat glandlike structure expressed CEA positive. Conclusion ES cellderived epidermal stem cell Related Articles | Metrics

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A COMPARATIVE STUDY OF CIRCULATING FIBROCYTES AND FIBROBLASTS 2007, 38 (3):  300-303.  doi: Abstract ( )   Objective To compare the morphological characters and collagen synthesis between circulating fibrocytes and fibroblasts. Methods Circulating fibrocytes were isolated from human peripheral blood by centrifugation over FicollPaque and cultured in vitro. Then the collagen secreted into the medium was detected by the measurement of the hydroxyproline concentration with enzymolysis, and the collagen accumulated on the cell layer was determined by the optical density at 540nm of picrosirius red staining. The contents of collagen were assessed by Western blotting. Results The morphological characteristics of circulating fibrocytes were varying when cultured in vitro and the collagen types Ⅰ and Ⅲ were synthesized and secreted into the medium.Conclusion Circulating fibrocytes can differentiate into fi Related Articles | Metrics

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PROMOTING EFFECTS OF SEROPHARMACOLOGICAL PLASTRUM TESTUDINIS ON THE EXPRESSION OF BONE MORPHOGENETIC PROTEIN 4 OF RAT MESENCHYMAL STEM CELL IN VITRO 2007, 38 (3):  304-309.  doi: Abstract ( )   Objective To observe the effect of seropharmacological plastrum testudinis on the expression of bone morphogenetic protein4 (BMP4) of rat mesenchymal stem cells (MSCs) in vitro. Methods Mesenchymal stem cells were dissociated from rat bone marrow and were marked by Brdu and the expression of CD44. The growth of rat mesenchymal stem cells with low, middle and high seropharmacological plastrum testudinis were observed separately by means of morphological apperances. The locations of the BMP4 protein and BMP4 mRNA positive cells were detested by immunohistochemistry and fluorescent immunohistochemistry combined with laser confocal scanning microscopy and in situ hybridization stained by DAB. The expression quantity of BMP4 mRNA and BMP4 protein was detected by real time PCR and ELISA. Results Seropharmacological plastrum testudinis promoted the numbers of BMP4 and BMP4 mRNA positive cells and the expressions of BMP4 protein and BMP4 mRNA in a time and dosedependant manner; there were significant differences in comparison with control groups. Conclusion Promoting effect of seropharmacological plastrum testudinis on the prolife Related Articles | Metrics

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LEUKEMIA INHIBITORY FACTOR GENE TRANSFECTED HUMAN EMBRYONIC LUNG FIBROBLASTS FEEDER LAYER FOR EMBRYONIC GERM CELLS 2007, 38 (3):  310-314.  doi: Abstract ( )   Objective To establish an animalfree culture system for human embryonic germ cells. With leukemia inhibitory factor(LIF) genetransfected human embryonic lung fibroblasts as the feeder layer. Methods Eukaryotic expression vector pcDNA3.1(+)-LIF was transfected into human embryonic lung fibroblasts. LIF positive cells were obtained after being screened by G418 and identified by RT-PCR and Western blotting. Primordial germ cells(PGCs) of 5 to 9weekpostfertilization embryos were plated onto the transfected human embryonic lung fibroblast feeder cells, and cultured in a medium without hrLIF. The cell colonies derived from PGCs were also identified. Results RT-PCR and Western blotting results showed that the transfected cells expressed LIF. Primordial germ cells that was growing on the feeder layer formed typical multicellular colonies. The colonies were found to be strongly positive in AP activities, and expressed stage specific antigens SSEA-1, SSEA-4, TRA-1-60, TRA-1-81. Oct-4 expression was positive by RT-PCR.Conclusion LIF genetransfected human embryonic lung fibroblasts as the feeder l Related Articles | Metrics

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P>THE REGWATION OF GANGLIOSIDE ON PKC PATHWAYS AND ITS PROTECTIVE EFFECT ON SERUMDEPRIVED INJURY IN PC12 CELLS/P> 2007, 38 (3):  315-319.  doi: Abstract ( )   Objective To determinted whether GM1 had a protective effect on injury induced by serumdeprivation and the possible mechanism in PC12 cells. Methods The viability of PC12 cells was quantified by MTT after serumdeprivation. The number of apoptotic cells and necrotic cells were determined by Hoechst 33258/PI staining. And the change of PKC protein expression on PC12 cells’ membrane and cytosols was detected by Western blotting. Results 1The viability of PC12 cells decreased after serumdeprivation and the serum-deprivation for 24 hours was chosen as an injury model in this research. Most of the PC12 cells presented apoptosis 24 hours after serumdeprivation. In addition, the PC12 cells’ cytosols PKC protein decreased, while the PC12 cells’ membrane PKC protein increased significantly, and this result suggested PKC’s translocation to membrane and its activation. 2The viability of PC12 cells preincubated with GM1 in high concentrations(10,1,01μmol/L) increased significantly and GM1 protected PC12 cells from apoptosis after serumdeprived injury. GM1 reduced the damage of serumdeprivation on PC12 cells and inhibited PKC protein translocation after injury.3The repair fun Related Articles | Metrics

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THE EFFECT OF MACROPHAGECONDITIONED MEDIUM ON THE GROWTH OF PRIMARY MICE MYOBLASTS 2007, 38 (3):  320-324.  doi: Abstract ( )   Objective To investigate the effect of macrophageconditioned medium on the growth of mouse myoblasts. Methods After a 3day intraperitoneal injection with a mixture of muscle homogenate and starch, macrophages were obtained from mouse peritoneal cavity and cultured for 48 hours in serumfree DMEM/F12. Then the cultured medium was harvested. Myoblasts were obtained from newborn mice skeletal muscle and grown in DMEM/F12 supplied with 10% FBS for 3648 hours, and then exposed to 0.5% FBS medium with or without macrophageconditioned medium. Cells were harvested at 72 hours later, their growth was observed with Wright staining, LDH cytochemistry and immunocytochemistry. Results Macrophageconditioned medium can maintain the normal morphology and the activity of myoblasts grown in low concentration of serum and promote their proliferation. There was a significant difference between average absorbance and integral absorbance of LDH positive cells of the two groups(P<0.01 and P<0.000 1 respectively). The experiment group contained more positive cells compared with the control group, identified by immunocytochemistry for desmin.Conclusion Ma Related Articles | Metrics

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THE INDUCTION OF P19 CELLS INTO MYOCARDIUM CELLS USING 5-aza CYTIDINE EM>IN VITRO/EM> 2007, 38 (3):  325-329.  doi: Abstract ( )   Objective To analyze the features of P19 cells differentiating into cardiomyocyte after induced by 5-azacytidine(5-aza) in vitro. Methods The P19 cells were cultivated in the medium containing different concentrations(1,5 and 10μmol/L) of 5-aza in dishes laid with a thin layer of agar to form embryonic bodies(EBs). On the 7th day, EBs were transferred to coverslips in 24 well plates, allowed to adhere and be cultured in the medium without 5-aza. The beating of cells was observed. α-sarcomeric actin and cardiac Troponin T(cTnT) immunofluorescence stain and GATA-4, α-MHC gene RT-PCR analysis were used to identify cell differentiation. Results After the 5-aza induced EBs were cohesively cultured, spontaneously and rhythmically beating cells were found within the EBs outgrowths, which were α-sarcomeric actinpositive and cTnTpositive, and expressed GATA4, α-MHC gene simultaneously. The treatment with 10μmol/L 5-aza can increase the rate of cardiomyocyte differentiation.Conclusion The combina Related Articles | Metrics

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THE EFFECTS OF LASER IRRADIATION ON INTRACELLULAR ROS，CALCIUM CONCENTRATION AND CELL MEMBRANE INTEGRITY 2007, 38 (3):  330-333.  doi: Abstract ( )   Objective To investigate the effects of laser irradiation on intracellular ROS(reactive oxidant species), intracellular calcium concentration(［Ca2+］i，and cell membrane integrity in the process of live cell imaging with confocal laser scanning microscopy. Methods The effects of a given laser irradiation on ROS, intracellular calcium concentration (［Ca2+］i and cell viability were revealed respectively by stained ECV304 with H2DCFDA，Fluo4AM and calceinAM/PI, and visualized and analyzed using ultra view LCI(live cell image)confocal microscopy. Results The irradiation of 488nm laser induced fluorescent intensity of DCF to increase abruptly and attain the climax in about 80 seconds, afterwards the fluorescent intensity fell and returned to the baseline. In the 70 minutes of the irradiation, the fluorescent intensity of intracellular Fluo4 kept a slightly ascending tendency. The fluorescent intensity of calcein decreased 15minutes after the irradiation, and serval cells were PI positively stained.Conclusion 488nm laser irradiation Related Articles | Metrics

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A SCANNING ELECTRON MICROSCOPE STUDY ON THE IMMATURE DENDRITIC CELLS BEFORE AND AFTER THE ELECTRANSFECTION OF HUMAN HEPATIC CANCER CELL RNA 2007, 38 (3):  334-338.  doi: Abstract ( )   Objective To observe the morphologic changes of immature dendritic cells(imDCs) before and after the electransfection of human hepatic cancer cell RNA. Methods Monocytes were purified from human peripheral blood, and induced into imDCs. Then human hepatic cancer cell RNA was electransfected into monocytederived imDCs. ImDCs were identified by the immunocytochemical method with 7 specific antibodies before and after electransfection. These dendritic cells were observed by scanning electron microscopy. Results After electransfection of human hepatic cancer cell RNA there were few changes of molecule expressions in imDCs. ImDCs were in round, oval and irregular shapes before electransfection. Their sizes were not identical but all bigger than monocytes. There were many protrusions in different shapes which looked like dendrite, or/and bubble, veil cloud on the surface of these imDCs. Although there were some cell fusions and cell deaths after electransfection, most imDCs recovered from the damage. Electransfecting human hepatic cancer cell RNA into imDCs would make pores on cell membrane.Conclusions The pores on cell membrane make it possible that the exogenous material enters imDCs. This study can prove the possibility of electransfecting human hepatic cancer cell RNA into imDCs to make cancer vaccine, which provides a new way for tumor biologic therapy. Related Articles | Metrics

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EXPRESSION OF BMP-7 IN HEPATIC TISSUE IN HEPATIC FIBROSIS RATS INDUCED BY CARBON TETRACHLORIDE 2007, 38 (3):  339-342.  doi: Abstract ( )   Objective To explore the relationship between the expression of BMP-7 in the hepatic tissue and hepatic fibrosis in rat hepatic fibrosis induced by CCl-4. Methods Healthy adult male Wistar rats were randomly divided into the control group and the hepatic fibrosis group. The control group was subcutaneously injected with oily solution of 0.12ml per 100 grams of rat weight every monday and thursday; the hepatic fibrosis group was subcutaneously injected with 60% carbon tetrachloride oily solution of 0.3ml per 100 grams of rat weight every monday and thursday. Rats were continuously injected for 6 weeks, 10 weeks, 16 weeks and 21weeks respectively. When the experiment ended, all the rats were killed. The hepatic middle lobule was taken and embedded with paraffin. Collagen fibers in the hepatic tissue were detected by Sirius red staining. The expression of BMP-7 in the hepatic tissue was detected by immunohistochemical staining and Western blotting. Results In the rat hepatic tissue, there were few collagen fibers in portal veins and the portal area in the control group. Proliferation of collagen fibers could be observed in each rat of the hepatic fibrosis group and hepatic fibrosis was aggravated with the lasting of the experiment. In immunohistochemical staining, several BMP-7 positive cells were observed in the rat livers of the control group; many BMP-7 positive cells were observed in the 6-week group. The expression of BMP-7 positive cells gradually decreased after 10 weeks,and they were seldom seen in the 21-week group. The Western blotting results were basically the same as those of immunohistochemical staining.Conclusion In rat hepatic fibrosis induced by carbon tetrachloride, the expression of BMP-7 in the hepatic tissue was opposite to that in the hepatic fibrosis. BMP-7 can protect the hepatic fibrosis to some degree. Related Articles | Metrics

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THE ULTRASTRUCTURE OF HUMAN PERIODONTAL LIGAMENT CELLS AFTER THE OSTEOINDUCTIVE CULTURE 2007, 38 (3):  343-346.  doi: Abstract ( )   Objective To observe the ultrastructure of human periodontal ligament cells after the osteoinductive culture. Methods Human periodontal ligament cells(hPDLCs) were cultured in the osteoinductive medium with 10SUP>-7/SUP> mol/L dexamethasone, 50 mg/L α-scorbic acid and 10SUP>-2/SUP> mol/L β-glycerolphosphate or in ordinary culture for 14 days.Transmission electron microscopy was used to observe the ultrastructural changes. Results After osteoinductive culture,there were richer mitochondria,endoplasmic reticulum and Golgi complex in the cytoplasm of hPDLCs than that in the ordinary clutured hPDLCs.And lots of myelin like structures and matrix vesicles with middle or low electron density were observed in the hPDLCs cultured with the osteoinductive medium.Conclusion The ultrastructural characteristics of hPDLCs after the osteoinductive culture were similar to those of mineral-forming cells,such as osteoblast and c Related Articles | Metrics

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IMMUNOHISTOCHEMICAL STUDY OF ISLET PP CELLS DURING THE HEALING PROCESS OF EXPERIMENTAL GASTRIC ULCER IN RATS 2007, 38 (3):  347-350.  doi: Abstract ( )   Objective To explore the possible function and significance of pancreatic polypeptide during the healing process of experimental gastric ulcer in rat. Methods The immunohistochemical PAP method,morphometry and image analysis were applied to study the changes of the morphology, numerical density on area(N\-A) and mean grey degree of islet PP cells during the healing process of experimental gastric ulcer in rats. Results Compared with normal control group(NCG) and saline control group(SCG), the N\-A of PP cells markedly decreased, and the mean grey degree markedly increased(P<0.05) on the 6th day after the experimental gastric ulcer group(EUG). During the 10th and the 28th day after the process of experimental gastric ulcer, the N\-A of PP-positive cell increased,and the mean grey degree decreased(P<0.05),especially on the 10th day.The results of morphological observation and image analysis in pancreas islet PP cells were basically the same during the healing process of experimental gastric ulcer. Conclusion During the healing process of experimental gastric ulcer,the N\-A and mean grey degree of PP-positive cell both changed.It indicated that PP cells played a role in the modulating process of experimental gastric ulcer. Related Articles | Metrics

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THE EFFECT OF TRANSPLANTATION OF MOUSE PRIMORDIAL GERM CELLS ON THE ACUTE DAMAGED LIVER 2007, 38 (3):  351-355.  doi: Abstract ( )   Objective To investigate the milieu-dependent differentiation of primordial germ cells(PEGs) in the acute damaged liver microenviroment. Methods After PGCs were cultured and proliferated, these cells were labelled with 5-bromo-2-deoxyuridine(BrdU), then transplanted into the acute damaged liver by CCl\-4 through tail vein. Two and four weeks later, the liver was extracted and 10μm -cryostat continuous sections were obtained. The existing and differentiation of the transplanted cells were identified by immunohistochemistry, immunofluorescence double staining and histochemistry for BrdU and hepatic-specific ALB, and the glycogen. Results Transplanted PGCs were found to be incorporated into the acute damaged liver and differentiated into hepatocytes, compensating for acute liver failure.Conclusion PGCs can be induced to differentiate into hepatocytes in the acute damaged liver microenvironment, and can be used for cellular hepatoplasty to treat severe liver disease. Related Articles | Metrics

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THE ISOLATION, PURIFICATION AND IDENTIFICATION OF WISTAR RAT’S ISLET 2007, 38 (3):  356-359.  doi: Abstract ( )   Objective The experiment aims at probing the best condition of the isolation and purification of rat islets. Methods The islets were isolated from rat pancreas by hepatopancreatic duct perfusing with collagenase and purified with Ficoll 400 discontinuous density gradient centrifugation. Then the purified islets were subjected to histological staining, electron microscopy and radioimmunoassay for identification of specificity and viability. Results The histological staining revealed that the viability and the purity of the purified islets were above 95% and 85% respectively. Electron microscopy showed that the purified islets were morphologically intact with clear membrane and plenty of secreting granules. Radioimmunoassay demonstrated the secreted insulin concerntration between low-glucose groups and high-glucose groups varied significantly, which verified the good function of the islets.Conclusion Hepatopancreatic duct perfusing with collagenase is a good method for digestion. There are many factors that influence the quantity and quality of the acquired islets, such as the completed expansion of pancreas, the concentration and viability of collagenase and the digested time, etc. Related Articles | Metrics

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THE EFFECT OF ISCHEMIA-REPERFUSION ONMATRIX METALLOPROTEINASE-1 IN RAT HEART 2007, 38 (3):  360-364.  doi: Abstract ( )   Objective To investigate the effect of ischemia and ischemia/reperfusion(I/R) in rat heart on Matrix metalloproteinase-1(MMP-1). Methods The I/R animal models were established by shutting down and reopening the anterior interventricular branch with a silver clamp,then the distribution and amount of MMP-1 of the normal and I/R rat hearts were observed by immunohistochemical staining and Western blotting and analyzed by computer image analysis. BR>Results 1. Immunohistochemical staining showed MMP-1 existed mainly in the cardiac matrix.There were strong positive reactions in fibrocytes, smooth muscle cells of the blood vessel and endothelial cells of capillaries.MMP-1 didn’t show distinct changes 30 minutes after ischemia,while its concentration increased dramatically 60 minutes after ischemia.The positive reaction of MMP-1 increased 30 minutes after I/R,and 60 minutes after I/R there was large fusion areas in MMP-1 existing regions. 2. Quantitative analysis showed no dramatic changes of MMP-1 after ischemia for 30 minutes(P>0.05),while dramatic changes were seen 60 minutes after ischemia(P<0.05).MMP-1 changed dramatically 30 minutes and 60 minutes after I/R. 3. Western blotting showed that there were no distinct naked-eye-observable changes.The bands of MMP-1 became widened 30 minutes after I/R, and became obviously widened 60 minutes after I/R.Conclusion 1. MMP-1 is secreted by fibrocytes,smooth muscle cells and endothelial cells of cardiac tissue under physiological conditions,and cardiomyocytes has the potential to secrete MMP-1 under ischemia or I/R. 2. The longer time the heart ischemia lasts,the greater MMP-1 concentration will increase.Reperfusion can increase MMP-1 concentration to an even higher level,which may be the main cause of the collagen destruction after heart I/R. BR> Related Articles | Metrics

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THE LOCALIZATION AND EFFECT OF QUANTUM DOTS ON ULTRASTRUCTURE OF MOUSE ABDOMINAL CAVITY MACROPHAGES IN VITRO 2007, 38 (3):  365-368.  doi: Abstract ( )   Objective To observe the distribution and the effect of the quantum dots(QDs) on mouse abdominal cavity macrophages. Methods The QDs were co-cultured with mouse abdominal cavity macrophages in vitro. The differentiation and effect of the QDs on macrophage ultrastructures were observed under electronic microscope. Results The QDs were enveloped with unit membrane and internalized in the cytoplasm of the macrophage under transmission electron microscope.And it formed vacuolelike structures in the macrophage.There were many lamellar processes on the surface of the macrophage under scanning electron microscope.Conclusion The QDs can promote macrophage activation,and make its surface projection increased.The QDs were internalized by the macrophage, distributed in the cytoplasm, and formed vacuolelike structures enveloped with unit membrane. Related Articles | Metrics

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THE EFFECT OF GANODERMA SPORES ON THE EXPRESSION OF Cdk4 IN EMBRYONIC NEURAL TUBE DEFFECTS INDUCED BY RETINOIC ACID 2007, 38 (3):  369-372.  doi: Abstract ( )   Objective To explore the expression of Cdk4 of neural epithelia,when treated with ganoderma spores in embryonic neural tube defects(NTDs) induced by retinoic acid in pregnant mice. Methods When E7.75d, mice of the ganoderma spores and the control groups were given intragastrically onetime retinoic acid.Then the ganoderma spores solution was given intragastrically to the ganoderma spores group. At E10.5d,immunofluorescence histochemistry and Western blotting were applied to examine the expression of Cdk4. Results The expression of Cdk4 was detected in the neural epithelia of the normal embryos in the ganoderma spores group.The abundance of Cdk4 was no difference between the blank control group and the normal control group.Although the expression of Cdk4 was detected in the neural epithelia of the abnormal embryos in the ganoderma spores group,its level was obviously lower than that in the blank control group or the normal control group.Conclusion Ganoderma spores may promote the expression of Cdk4 in the neural epithelia of neural tube to reduce the occurrence of embryonic NTDs induced by retinoic acid in pregnant mice. Related Articles | Metrics

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APPLIED ANATOMY OF THE ARTIFICIAL COCHLEAR IMPLANTATION IN YOUNG CHILDREN 2007, 38 (3):  373-375.  doi: Abstract ( )   Objective To provide anatomic data for operation of inserting the electron cochlear in young children. Methods Fourteen heads,28 sides specimens of young children of 1-to-5-year old were dissected,through posterior tympanum approach,via mastoidectomy,posterior tympanoto to enter posterior tympanum.The related anatomy structures of the location of the electron cochlear inserted into the proper sites were observed and measured under surgical microscope. Results The round window was seated in superior part of the round window niche.The pyramidal eminence,tendo musculi stapedius,incudostapedial joint,base of stapes,cochleariform process,round window niche and promontorium tympani were all visible from different directions.The posterior arch of stapes was situated in the prozone of scala.Scala was situated in the posteroinferior scala vestibuli.The distance from the middle point of the anterior border of the round window niche to the inferior wall was (149±0.42)mm,to the posterior wall of the Scala tympani (0.90±0.31)mm,to the basal tissue (149±0.41)mm,to the pyramidal eminence (328±0.55)mm,to the lateral semicircular canal (741±0.90)mm,to the inferior margin of the base of stapes (309±0.53)mm.Conclusion It is considered that the location of the insertion should be at the middle point of anterior border of the round window niche anterior from 0.90mm to 149 Related Articles | Metrics

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DiI ANTEROGRADE-TRACING THE CORTICOSPINAL TRACT OF THE RAT 2007, 38 (3):  376-378.  doi: Abstract ( )   Objective To study the effect of anterograde-tracing the corticospinal tract of the rat using DiI a tracer. Methods Twenty-four Wistar rats were randomly divided into DiI group and degeneration group. 5% DiI were stained on the motorsensor cortex of the DiI group rats; the corresponding cortex of the degeneration group were destroyed.After 5d,10d,15d and 20d,the rats were killed respectively.The brains and spinal cords were sliced and observed. Results In the 5d DiI group,labelled fibers were only observed both in telencephalon and diencephalon,and in 10d,15d and 20d groups,corticospinal tracts were well labelled through the brain and spinal cord.In the degeneration group,black degenerating fibers of the spinal cords were well demonstrated,especially in the 5d group.However, above the spinal cords,no obvious degenerating fibers could be found.Conclusion It’s a simple,convenient and valuable method,using DiI as an anterograde tracer,to trace corticospinal tract,especially the part through midbrain to spinal cord. Related Articles | Metrics