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2007, Volume 38 Issue 5
06 October 2007 Previous Issue    Next Issue
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EFFECT OF THE ARTIFICIAL NERVE GRAFT ON NERVE REGENERATION IN RAT SCIATIC NERVE OF TWO MONTHS DEFECT MODEL 2007, 38 (5):  505-510.  doi: Abstract ( )   Objective The aim of this study was to evaluate the effect of bridging a gap in rat sciatic nerve of 60-day defect with artificial nerve graft. Methods Left sciatic nerves of 20 SpragueDawley rats were transected at the midthigh level to form nerve defect. After two months,eight rats with artificial nerve graft bridged defect were used as the experiment group.Autologous nerve grafts were used as the positive control in six rats, and another six rats keep defect as the negative control. Three months after the operation, a combination of electrophysiology examination, gastrocnemius muscle wet weight, histological assessment including light microscopy, transmission electron microscopy and immunohistochemistry were utilized to evaluate the nerve repair effects. Results In the artificial nerve graft group, the motor nerve conduction velocity, gastrocnemius muscle wet weight, the average thickness of regeneration myelin sheath and G ratio were similar to those of the autologous nerve graft group. Conclusion Artificial nerve graft can be used for bridging a gap in 60day nerve defec Related Articles | Metrics

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EXPRESSION OF δ-CATENIN IN THE CEREBELLUM OF RAT WITH ESTROGEN REPLACEMENT 2007, 38 (5):  511-514.  doi: Abstract ( )   Objective To investigate the expression of δ-Catenin in the cerebellum of ovariectomed rat with estrogen replacement and the estrogen effect on δ-Catenin expression in the cerebellum of the rats with ovariectome. Methods The rats were divided into normal group, sham operation group, estrogen replacement treatment group and placebo group. Then the rats were artificially injected estradiol or placebo after ovariectome. The expression of δ-Catenin in the cerebellum was detected by immunohistochemical method and was quantitated by Western blotting analyses. Results Immunohistochemical method and Western blotting analyses both displayed the expression of δ-Catenin in all the groups, while the expression in placebo group was the lowest among the four groups, the expression of δ-Catenin in the groups of rats with estrogen replacement treatment and sham operation was significantly higher than that of placebo group.Conclusion The estrogen may upregulate the expres Related Articles | Metrics

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ISOLATION OF FOREIGN NEURONS IN CORNU ANTERIUS MEDULLAE SPINALIS AFTER ARTIFICAL SOMATICAUTONOMIC NERVE ANASTOMOSIS OF RATS AND STUDY OF EXPRESSION CHANGES OF AGRIN 2007, 38 (5):  515-519.  doi: Abstract ( )   Objective To study the changes of agrin expression in spinal cord motor neurons after somaticautonomic ventral root (VR) anastomosis and somaticsomatic VR anastomosis, and the factors related to the synapse formation of foreign nerve regeneration. Methods Artifical somaticautomomic reflex pathway and somatic nerve selfanastomosis were established in rats. Retrograde tracing was carried out to label the motor neurons, and the neurons were isolated by flow cytometry. Realtime PCR was used to investigate the agrin mRNA expression of the neurons.Anterograde tracing and immunofluorescence technique were employed to label the new synapses of pelvic ganglia (PG). Results After 4 months of surgery, the somatic nerve can regenerate into PG successfully and form new synapses observed under laser scanning confocal microscope. The neurons involved in foreign nerve regeneration can be isolated successfully by retrograde tracing and flow cytometry.The expression of agrin in LSUB>4/SUB>-LSUB>6/SUB> VRs anastomosis group was lower than that of the control group of LSUB>4/SUB>-LSUB>4 /SUB>VR self-anastomosis (P<0.05) and the normal group (P<0.05).Conclusion The somatic nerve could regenerate into PG and form new synapses.The differences of agrin expression Related Articles | Metrics

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THE THERAPEUTICAL EFFECTS OF YANGXUEQINGNAOKELI ON HIPPOCAMPUS CA1 NEURONS IN MONGOLIAN GERBILS AFTER GLOBAL ISCHEMIC REPERFUSION INJURY 2007, 38 (5):  520-524.  doi: Abstract ( )   Objective To investigate the therapeutical effects of Yangxueqingnaokeli(YXQNKL) on global ischemia reperfusion injury in Mongolian gerbils. Methods The global cerebral ischemia of gerbils was induced by clamping bilateral carotid arteries for 30 minutes and reperfused for 5 days. Nissl staining was employed to observe the morphology and numbers of the gerbils’ CA1 hippocampal neurons, and immunohistochemical stainings of Glusyn, NCS-1 and caspase-3 were used to observe their expressions in the gerbils’ CA1 hippocampal neurons. Results Nissl staining showed that the CA1 hippocampal neurons both in I/R+0.4g/kg YXQNKL group and I/R+0.8g/kg YXQNKL group were much more than those in the I/R group (P<0.05), and immunohistochemical stainings indicated that the positive cells of NCS-1 and caspase-3,Glusyn in CA1 hippocampal neurons were significantly decreased compared with those in the I/R group (P<0.05).Conclusion Treatment with YXQ can increase the living neurons in hippocampus CA1 region potently against global ischemia reperfusion brain injury,which is closely related to the decreased expression of NCS-1,caspase-3 and Glusyn. Related Articles | Metrics

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EXPRESSION OF GFAP IN RAT HYPOTHALAMUS AFTER DIFFERENT TEMPERATURE HEAT STRESSES 2007, 38 (5):  525-527.  doi: Abstract ( )   Objective To investigate the expression of GFAP in rat hypothalamus after acute heat stress. Methods The rats were caged in a experimental incubator for 60 minutes, the temperature within the incubator was adjusted to 24℃,34℃,38.5℃ or 42℃, the humidity was 60%. Single anti-GFAP immunohistochemical (ABC) method and antiFos and GFAP double immunohistochemical method were used to observe the expression of GFAP in hypothalamus in different ambient temperatures after heat stress. Results The GFAPpositive cells were rare in hypothalamus at 24℃, however it was increased in many nuclei(anterior hypothalamic area, paraventricular nucleus,arcuate nucleus, suprachiasmatic nucleus and supraoptic nucleus)at 34℃ and peaked when ambient temperature was 38.5℃, and then decreased. However Related Articles | Metrics

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PROTEOMICS ANALYSIS OF THE ANTERIOR LOBE, THE POSTERIOR LOBE AND THE PARAFLOCCULUS IN MOUSE CEREBELLUM 2007, 38 (5):  528-531.  doi: Abstract ( )   Objective To study the different expressions of proteins in different regions of cerebellum. Methods Proteins isolated from the anterior lobe, the posterior lobe and the paraflocculus of cerebellum were separated by twodimensional (2D) gel electrophoresis. Using PDQuest software, proteins on the 2D maps were selected and further analyzed by matrixassisted laser desorption /ionization twostage time of flight (MALDI TOF/TOF) tandem mass spectrometry. The mass spectrometric data were used to identify the proteins through the general proteomics standard (GPS) search and Mascot database. Results There were five proteins identified in three regions of cerebella. The level of pyruvate kinase isozyme M2 was higher in the anterior and posterior lobes of cerebellum than in the paraflocculus. Heterogeneous nuclear ribonucleoprotein L and eukaryotic translation elongation factor 2 were only identified in the posterior lobe with low expression, and were not detected in other regions. The expressed sequence AI429145 was only detected in the posterior lobe. The paraflocculus expressed higher level of ATPase (H+ transporting, VO subunit D, isoform 1) compared with the anterior and posterior lobes.Conclusion The different expressions of those proteins may be important for the understanding of the specific functions Related Articles | Metrics

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BETA AMYLOID PEPTIDE 25-35 INDUCES Tau PROTEIN HYPERPHOSPHORYLATION IN CORTICAL NEURONS THROUGH JNK/p38 MAPK PATH IN RAT EMBRYO 2007, 38 (5):  532-536.  doi: Abstract ( )   Objective To investigate the effect and the molecular mechanism of aggregated beta-amyloid peptide 25-35 (Aβ SUB>25-35/SUB>) on the level of Tau protein phosphorylation in rat embryo cortical neurons. Methods Western blotting and immunocytochemical stain were performed to observe the Tau protein phosphorylation and the expression of JNK/p38 MAPK Results The level of Tau protein phosphorylation in the sites of SerSUP>396/SUP>, SerSUP> 199/202/SUP> and ThrSUP>205/SUP> increased after Aβ SUB>25-35/SUB> of 20μmol/L was exposed to cortical neurons, meanwhile the level of JNK/p38 MAPK also increased after treatment with Aβ25-35 for 12 hours. Pretreatment with specific inhibitor of JNK/p38 MAPK markedly attenuated Tau pr Related Articles | Metrics

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INHIBITION OF NOGO-A GENE EXPRESSION BY RNA AND ITS EFFECTS ON DOPAMINE RELEASE IN PC12 CELLS 2007, 38 (5):  537-541.  doi: Abstract ( )   Objective To study the effect of Nogo-A gene silence on dopamine release in PC12 cells. Methods The small hairpin RNA(shRNA) eukaryotic expression vector targeting Nogo-A gene was constructed and transfected into cultured PC12 cells by lipofecamine2000. The inhibiting effect was detected by semiquantitative reverse transcription(PCR) and Western blotting analysis. The dopamine release was detected by HPLC(high performance liquid chromatography) after transfection. Results The Nogo-A gene expression was inhibited and the dopamine release decresed significantly 48 hours after transfection.Conclusion The reasults suggest that Nogo-A be involved in the mechanism of dopamine realese in PC12 cells. Related Articles | Metrics

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DIFFERENTIATION OF EMBRYONIC STEM CELLSR1 INTO INSULIN PRODUCING CELLS 2007, 38 (5):  542-545.  doi: Abstract ( )   Objective To define the best culture conditions of ES-R1 and explore the possibility and feasibility of differentiation of ES-R1 to IPCs in vitro with various of factors. Methods ESCs were cultured on the mitomycin Ctreated MEF feeder layer cells. The culture medium of ES-R1 was supplemented with 10μg/L LIF. ESCs were transferred into the ultralow adherent dishes in the absence of LIF and feeder layers for the formation of EBs. On day 4 of EBs formation, EBs were transferred to collagen-Coated dishes and serumfree ITSA medium, pancreatic proliferation medium and pancreatic differentiation medium were added step by step, each for 6 days. During the differentiation from ESCs to IPCs, morphological methods, immunofluoresence staining, alkaline phosphatase staining, dithizong staining and insulin release test were used. Results Undifferentiated state was maintained by feeder cells and LIF. ESCs could form nested colonies with clear edge and smooth surface. Upon withdrawal of LIF and feeder layer, ESCs spontaneously formed into EBs in suspension. At the end of induction of differentiation, the three dimensional IPCs were formed. The induced IPCs were found to be stained crimson red by DTZ, insulin and glucagon immunohistochemistry staining positive, insulin release test positive.Conclusion Combination of LIF and MEF feeder layer culture may maintain the proliferation of ES cells without differentiation. Various factors used at different times, ES-R1 can differentiate into IPCs which possess ch Related Articles | Metrics

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THE IMPLICATION OF PLC-γ2 IN GASTRIC CANCER MGC80-3 CELLS TREATED BY TPA 2007, 38 (5):  546-551.  doi: Abstract ( )   Objective To study the implication of phosphoinositidespecific phospholipase Cγ2(PLC-γ2) in gastric cancer MGC80-3 cells treated by TPA. Methods The effect of 12Otetradecanoylphorbol1, 3acetate(TPA) on gastric cancer MGC80-3 cells was detected and analyzed by DAPI staining under fluorescence microscope. The effect of TPA on the expression level of PLC-γ2 protein was detected by Western blotting, when nuclear and cytoplasmic protein fractions were prepared through lysis of cell and centrifugation. Localization and translocation of PLC-γ2 were observed under laser scanning confocal microscope with immunefluorescence technique. After MGC80-3 cells were pretreated by U73122(PLC-γ2 inhibitor), the effect of TPA on PLC-γ2 was detected by Western blotting and laser scanning confocal microscope with immunefluorescence technique, and the effect of U73122 on MGC80-3 cells followed by TPA was observed and analyzed by DAPI staining under fluorescence microscope. Results TPA induced the apoptosis of gastric cancer MGC80-3 cells. Meanwhile, TPA increased the expression level of PLC-γ2 protein and induced its translocation from cytoplasm to nucleus. U73122 inhibited the effect of TPA on the expression level of PLC-γ2 protein, but it did not affect on the cell apoptosis induced by TPA. However, the translocation of PLC-γ2 protein induced by TPA was not inhibited by its inhib Related Articles | Metrics

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STUDY ON GENETIC INSTABILITY OF nm23H1 GENE IN CHINESE WITH SQUAMOUS CELL LUNG CARCINOMA 2007, 38 (5):  552-557.  doi: Abstract ( )   Objective To evaluate microsatellite instability(MSI) and loss of heterozygosity(LOH) of the repetitive untranslated 5’region of the nm23H1 gene and their influence on the expression of nm23H1 in Chinese with squamous cell lung carcinoma(SLC). Methods Techniques such as DNA extraction from tissue and fluorescencebased polymerase chain reaction singlestrand conformation polymorphism(FPCR-SSCP) combined with an automated DNA sequencer were used to study MSI and LOH of nm23H1. Immunohistochemistry and LeicaQwin computer imaging techniques were used to assess the expression of nm23H1 gene. Results Of these cases of informative tumors 44 of 50, the frequencies of MSI, LOH and nm23 H1 protein positive were 11.36%, 25.00% and 50.00% respectively. The frequencies of MSI were 8.70%， 10.00% and 18.18% in TNM stage Ⅰ, Ⅱ and Ⅲ respectively; The frequencies of LOH were 30.43%, 20.00% and 18.18% in TNM stage, Ⅰ, Ⅱ and Ⅲ respectively and the frequency of MSI and LOH were 10.00% and 20.00% in lymph node metastasis cases compared with 12.50% and 29.17% in those without metastasis. However, the frequencies of MSI and LOH showed no correlation with TNM stage and lymph node metastasis(P>0.05). All the 5 cases with MSI positive belonged to moderate (G2) or good(G1) histological grade and all were alive in a year, but the frequency of MSI was not related to tumor differentiation and survival status of patient(P>0.05). The positive frequency of nm23H1 protein in lymph node metastasis cases (30.00%) was significantly lower than those without metastasis(66.67%)(P<0.05). TNM stage Ⅲ(18.18%) also exhibited lower positive frequency of nm23H1 protein compared with stage Ⅰ(65.22%)(P<0.05). Furthermore, the frequencies of MSI and LOH were not related to the expression of nm23H1 protein(P>0.05). There was also no difference in nm23H1 protein expression intensity analyzed by computer imaging(P>0.05). Conclusion The increase in the amount of nm23H1 protein expression can play an important role in restraining metastasis of squa Related Articles | Metrics

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EFFECTS OF HUMAN INSULINLIKE GROUTH FACTOR1 ENGINEERING CELLS ON DIABETES MELITUS 2007, 38 (5):  558-563.  doi: Abstract ( )   Objective To investigate the decreasing blood glucose effects of transplanting human insulinlike growth factor1(hIGF-1) engineering cells to diabetic model mice. Methods The hIGF-1 cDNA fragment was cloned into pAAV vector, then the viral particles were packaged with HEK293 incasing cells and the virus titer was identified with HT1080 cells. The marrow stromal cells were infected with high virus titer supernatant and transplanted into the diabetic model mice abdominal cavity. The effects of transplanting of virus infected marrow stromal cells on blood glucose of diabetic model mice were detected. Results The pAAV/ hIGF-1 plasmid was successfully constructed and the viral titer is up to 1012. The infection rate of marrow stromal cells was 60%. After the diabetic model mice were transplanted with bone marrow stromal cells that stably express hIGF-1 gene, the blood glucose decreased obviously (P0.05).Conclusion The marrow stromal cells that stably express hIGF-1 gene can obviously decrease blood glucose of diabetic model mice, which suggest that hIGF-1 gene can b Related Articles | Metrics

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EFFECTS OF SIMVASTATIN ON OSTEOBLAST VEGF EXPRESSION AND MICE CALVARIA ANGIONENESIS 2007, 38 (5):  564-568.  doi: Abstract ( )   Objective To explore the mechanism of osteogenesis from statins through observing the effects of simvastatin on the VEGF expression of calvaria osteoblasts and its local influence on the mice calvaria. Methods Rat calvaria osteoblasts were harvested and cultured. Cells were treated with simvastatin for 24 hours. RT-PCR and immunocytology were performed to detect the expression of VEGF. Simvastatin was injected subcutaneously over the calvaria of mice for 4 days, the calvaria were pathologically examined. Results VEGF expression in rat calvaria osteoblasts was promoted by simvastain at both transcriptional and translational levels. In vitro the calvaria thickness and angiogenesis were increased by local administration of simvastatin. Conclusion Simvastatin can promote osteoblasts VEGF expression in vitro and mice calvaria angiogenesis and in Related Articles | Metrics

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INDUCTION OF APOPTOSIS BY EXTRACT OF TOTAL FLAVONOIDS OF CHRYSANTHEMUM INDICUM ON ADJUVANT ARTHRITIS SYNOVIAL CELLS 2007, 38 (5):  569-571.  doi: Abstract ( )   Objective To study the effect of the extract of total flavonoids of chrysanthemum indicum(TFC) on adjuvant arthritis synovial cells. Methods 0.1ml of the complete Freund’s adjuvant was subcutaneously injected into the right hind feet pads of the SD rats. 24 days after immunity synovial cells in knee joint were treated with TFC and inhibition of proliferation was measured with MTT assay. DNA fragmentations were analyzed with DNA gel electrophoresis. Fluorescence staining of Hoechst 33258 to observe apoptotic body. Results The IC50 of TFC on synovial cells was 112mg/L. DNA gel electrophoresis showed ladderlike strap. Apoptotic bodies were observed by Hoechst 33258.Conclusion TFC can inhibit proliferation and induce apoptosis in synovial cells, and exerts therapeutical effect on rheumstoid arthritis. Related Articles | Metrics

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THE DISTRIBUTION， CLONING AND ANALYSIS OF PARTIAL SEQUENCE OF LH AND ITS RECEPTOR IN SUBMAXILLARY GLAND OF RATS 2007, 38 (5):  572-576.  doi: Abstract ( )   Objective To study the localization of luteinizing hormone(LH), its receptor (LHR) and their mRNA in submaxillary gland of rats, and to provide a theoretic evidence for further studying functional significance of the LH in submaxillary gland. Methods Immunohistochemical methods and in situ hybridization methods were used in the experiment. After isolation of the total RNA from the submaxillary gland, RT-PCR was conducted to obtain LH and LHR cDNA by using the degeneration primers. The products of PCR were analyzed by sequencing with Sanger’s method, respectively. Results The epithelial cells of serous acinus in submaxillary gland of rats showed LH and LHR immunoreactivity, which were located in cytoplasm with negative nuclei. LH and LHR mRNA hybridized signals were also detected in cytoplasm in the above cells. LH cDNA sequence was identical to that of the LH which has been reported in rat pituitary, and LHR cDNA sequence was identical to that of the LHR which has been reported in rat testis.Conclusion The epithelial cells of serous acinus in submaxillary gland of rats may not only produce LH but also be its target cells. LH may be involved in the functional regulation of submaxillary gland through autocrine o Related Articles | Metrics

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EFFECTS OF HEROIN DEPENDENCE ON Gas, SS POSITIVE CELLS IN GASTRIC ANTRUM OF RATS 2007, 38 (5):  577-580.  doi: Abstract ( )   P>Objective To explore the effects of heroin dependence on the gastrin(Gas), somatostatin(SS) positive cells in gastric antrum of rats and comprehend the possible functions of Gas, SS during heroin dependence. Methods Immunohistochemical SABC method and imagine analysis technique were employed. Results 1.Compared with the normal control group(NCG) and saline control group(SCG), the number of Gas positive cells increased (P<0.05), the intensity of staining stepped up gradually with the passage of time during dependence. The results of imagine analysis showed that the mean grey degree of Gas positive cells had the trend of descent, and it was lower than that in the NCG and SCG with the lowest degree on the 38th day. 2. The intensity of staining and the number of SS positive cells in gastric antrum in heroin dependence group(HDG) increased obviously compared with that in NCG and SCG(P<0.05) during heroin dependence, the mean grey degree stepped down remarkably than that in NCG and SCG.Conclusion The Gas, SS positive cells in gastric antrum of rats participate in the regulation course during heroin dependence by increasing the cell number and enhancing Gas and SS synthesis./P> Related Articles | Metrics

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IMPROVED ISLET SURVIVAL WITH RAT EMBRYONIC FIBROBLAST COCULTURE 2007, 38 (5):  581-584.  doi: Abstract ( )   Objective Islets transplantation of pancreatic islets has been demonstrated to be a viable alternative to exogenous insulin therapy for diabets mellitus. However, it requires that the islets must have a good function. This study was design to investigate how to retain best the islet function in transplantation as cocultured with EFCs. Methods The EFCs were isolated from SD rat fetus and islets were purfied from SD rats. The viability of the isolated islets was assessed by AO/PI double staining. The islets were cultured alone(group Ⅰ), coculture with EFCs(group Ⅱ) and EFCs only(group Ⅲ). Islet viablity was assessed by mitochondrial activity and insulin release assay. Results Morphology assay: Islets in group Ⅱ exhibited excellent morphology, with greater than 95% staining positive as detected by AO/PI. Insulin release assay showed that: There was significantly a higher simulation indece(SI) in group Ⅱ as compared with group Ⅰ(P<0.05) except the first day. Mitochondrial activity: A total of the absorbance of the islet cells by MTT in 3,5,7,14 day between group Ⅰ and group Ⅱ showed significant difference(P<0.05, or P<0.01).Conclusion This preliminary series of controlled experiments suggest that cocultrue of Related Articles | Metrics

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IMMUNOHISTOCHEMICAL STUDIES ON ENDOCRINE CELLS IN THE DIGESTIVE TRACT OF LAIWU BLACK GOAT 2007, 38 (5):  585-588.  doi: Abstract ( )   Objective The localization and morphology of somatostain(SS),serotonin(5-HT),gastrin(Gas) and vasoactive intestinal peptide(VIP) immunoreactive cells in Laiwu black goat digestive tract were studied. Methods Immunohistochemical SABC method was used. Results The morphology of endocrine cells in digestive tract was of multiplicity.The density of 5-HT cells was the highest in the colon,but not found in the esophagus,cardia gastric and cecum.The results showed that SS cells were distributed in the pylorus mostly,but not detected in the cardia,colon,cecum and rectum.Gas positive cells were the most in the duodenums,while in the esophagus and cardia was not discovered.In the cardia VIP cells were maximum,but in the esophagus,duodenums and ileum did not appear.Conclusion The location of the SS-,5-HT-,Gas- and VIP-positive cells mostly show up in the colon,the pylorus,the duodenums and the cardia respectively Related Articles | Metrics

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RELATIONSHIP BETWEEN EXPRESSION OF SENESCENCE MARKER PROTEIN30 AND LIVER DISEASES 2007, 38 (5):  589-592.  doi: Abstract ( )   Objective To detect the expression of SMP-30 protein in normal liver, virus hepatitis, hepatic cirrhosis, HCC and their corresponding paracancerous tissues, so as to study the relationship between SMP-30 and liver diseases. Methods The MBP-SMP-30 fusion protein was expressed and purified from expression plasmid that contained 165 amino acids of SMP-30 Cend. The fusion protein was used to immunize rabbit to prepare the antiMBP-SMP-30 polyclonal antibody. Thirty normal liver, 10 virus hepatitis and 49 hepatic cirrhosis samples, 48 cases of HCC and their corresponding paracancerous tissues were used for detection. Immunohistochemical staining for SMP-30 was performed with SABC method on paraffin sections. Results SMP-30 was found in the cytoplasm and nucleus of hepatocyte. The protein expression level in HCC paracancerous tissues was higher than that in others (P<0.05). The expression of SMP-30 protein in HCC was no correlation with tumor size and pathological grade. Conclusion SMP-30 in hepatocytes can be detected with polyclonal antibody of MBP-SMP-30 fusion protein. SMP-30 is associated with progression of liver diseases. The higher expression of SMP-30 protein may be an early and protective event in HCC. Related Articles | Metrics

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CHANGE OF THE MITOCHONDRIAL DISTRIBUTION IN HUMAN OOCYTES BEFORE AND AFTER IN VITRO MATURATION 2007, 38 (5):  593-596.  doi: Abstract ( )   Objective To clarify the relationship between oocyte maturation and mitochondria distribution by investigating the distribution of mitochondrial in immatured and matured human oocytes. Methods Both immatured and matured oocytes were stained by Mito Tracker Green FM. After they were fixed by paraformaldehyde, the distribution of mitochondria was observed using laser scanning confocal microscope. Results Three mitochondria distribution patterns were identified: peripheral, semiperipheral and evenly diffused. 64.10%(50/78) of the GV oocytes presented a peripheral distribution of mitochondria. 45.16%(28/62) of the MI oocytes had maintained the peripheral distribution,while 38.71%(24/62) presented a diffused status. After in vitro maturation, 75.47%(80/106)of the oocytes displayed an evenly diffused type of distribution. Compared with the oocytes matured in vitro,the most marked feature of oocytes matured in vivo was that mitochondria distributed in the central region. Staining in the center of the oocyte was stronger than that in the peripheral region. Conclusion There were obvious changes in the distribution of mitochondria in human oocytes before and after maturation.The chief distribution pattern of mitochondria was changing fro Related Articles | Metrics

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THE EXPRESSION OF PECAM-1 IN RAT COLON ADENCARCINOMA AND LYMPHATICS 2007, 38 (5):  597-600.  doi: Abstract ( )   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 Related Articles | Metrics

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THE EFFECT OF SOYBEAN ISOFLAVONE ON LIPID PEROXIDATION AND LIVER ULTRASTRUCTURE IN OVARIECTOMIZED RATS 2007, 38 (5):  601-605.  doi: Abstract ( )   Objective To study the effects of soybean isoflavone (SI) on lipid peroxidation and liver ultrastructure in ovariectomized rats. Methods Seventy female SpragueDawley(SD) rats were randomly divided into 7 groups according to the levels of total cholesterol(TC) in serum: hyperlipoid group, estrogen group, lowdose SI group, middledose SI group, highdose SI group, sham group and normal control group. After bilateral ovaries were extirpated except sham and normal control groups for a week, the estrogen, different doses of SI or deionized water were fed with intragastric administration for 12 weeks. The rat body weight was weighted once per week and blood samples were collected at the time of ovariectomization, the 4th and 8th weeks after administeration at the time of being killed. The serum TC, triglyceride(TG), high or low density lipoprotein-Cholesterol(HDL-C, LDL-C), lipoprotein(a) and antioxygen enzyme were assayed. Results LDL-C levels in SI intervention groups were significantly lower than that in hyperlipoid groups but higher than that in normal control group; the levels of lipoprotein(a) were changed; there was almost no effect on HDL-C in serum. SI could retain integrity and ultrastructure of hepatocyte; there was similar structure in high dose SI group and normal control group. An obvious damage was detected in hyperlipoid group. Conclusion SI can improve lipoprotein(a) and lipid peroxidation in ovariectomized rats. A continuous intervention w Related Articles | Metrics

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A DERMATOGLYPHIC STUDY OF THE KEJIA PEOPLE IN TAIWAN OF CHINA 2007, 38 (5):  606-609.  doi: Abstract ( )   Objective To report hand dermatoglyphics parameters of the Kejia people in Taiwan of China. Methods Fingerprints and palmar prints were taken with informed consent.The sample comprised 100 males and 100 females. Results The results were obtained from TFRC,a-bRC,atd, tPD,fingerprint,hand thenar,hand interdigital,hand hypothenar,simian line.Conclusion This study is the first comprehensive dermatoglyphic research of Kejia people,and its dermatoglyphic database will be useful for future studies in anthropology,genetics and medicine. Related Articles | Metrics

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ISOLATION AND CULTURE OF FETAL MURINE EPIDERMAL STEM CELLS IN VITRO AND rAAV2/eGFP GENETIC TRANSFECTION 2007, 38 (5):  610-613.  doi: Abstract ( )   Objective To isolate,culture and identify epidermal stem cells(ESCs). The gene of enhanced green fluorescent protein(eGFP) was introduced via recombinant adeno-associated virus(rAAV) infection into the epidermal stem cells in vitro and the transfection efficiency under various tite of culture was determined. Methods 1. Getting fetal Wistar rats’dissociated single epidermal cells.2. Making laminin and CollagenⅣ the substitution of basal memrane,ESCs were isolated by adhering to murine laminin and CollagenⅣ.3. Epidermal stem cells were cultured in twenty-four-well plate,and cells number was controled 5×１０4 or so.After epidermal stem cells adhered to plate,diluted rAAV2/eGFP virus fluid with serum-free medium.The diluted rAAV2/eGFP virus fluid was added to the twenty-four-well plate according to the different MOI(viru gene/cells).The number of green fluorescence cells were counted under fluorescence microscope.The transfection efficiency was determined. Results 1. ESCs had better adhesive ability to laminin and higher colony formation efficiency(CFE) than that of keratinocytes.2. ESCs wer strongly positive with immunocytochemical staining of integrin β1 and keration 19(K19).3. After rAAV2/eGFP genetic transfection of ESCs,the positive cloning expressed highly eGFP gene along with time.Conclusion 1. These results suggest that rats’epidermal stem cells could been successfully isolated and enric Related Articles | Metrics

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CULTIVATION OF RETINAL PROGENITOR CELLS OF DIFFERENT AGES 2007, 38 (5):  614-617.  doi: Abstract ( )   P>Objective To study the cultivation of retinal progenitor cells of different ages. Methods Retinal progenitor cells of E14 and E18 SD rats were isolated,cultivated in suspension in modified DMEM/F12 serum-free medium and then differentiation was induced in vitro. Cells were observed under phase-contrast microscopy daily and identified by immunocytochemistry, scanning electron microscopy and transmission electron microscopy. Results Cultivated in DMEM/F12 serum-free medium,retinal progenitor cells formed cell spheres.After being plated,isolated cells migrated outwards and differentiated.Scanning electron microscopy demonstrated the morphology of the cell spheres and differentiated cells.Transmission electron microscopy demonstrated that there were stem cell-like cells in cell spheres and neuron- and glia-like cells after the plating.Immunocytochemistry demonstrated that most cells in spheres expressed neural stem cell marker nestin and cell division marker BrdU.After the plating, retinal progenitor cells could be induced to differentiate into various retinal cells,including Thy1 1-positive retinal ganglion cells.The percentage of retinal ganglion cells was 16.91%±4.05% at E14 and 4.65%±1.88% at E18.The differences were statistically significant(t=15.04,P<0.001).Conclusion Retinal progenitor cells can be cultivated successfully in modified DMEM/F12 serum-free medium.The cultivated retinal progenitor cells have the potential to proliferate freely and multi-differentiate.Early retinal progenitor cells are inclined to differentiate in Related Articles | Metrics

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EXPRESSION OF LEUKEMIA INHIBITORY FACTOR IN UTERUS IN THE PROCESS OF EMBRYO IMPLANTATION IN RAT 2007, 38 (5):  618-620.  doi: Abstract ( )   Objective To investigate the expression of cytokine leukemia inhibitory factor(LIF) in the process of embryo implantation in rat uterus. Methods The immunohistochemical ultra sensitive SP method was used to detect the expression of LIF protein in uterus. Results The expression of LIF was observed in uterine luminal epithelial cells,glandular epithelial cells,stromal cells,muscle cells and vascular endothelial cells;furthermore,immunoreactivity was stronger.Conclusion LIF plays important roles in the process of implantation of earlier embryo. Related Articles | Metrics

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THIN SECTIONAL ANATOMY AND HRCT OF FACIAL RECESS AND ITS ADJACENT STRUCTURES 2007, 38 (5):  621-624.  doi: Abstract ( )   Objective To study the thin section anatomy and HRCT of facial recess and its relative structures. Methods Thin sections were compared with HRCT scans.Facial recess and its relative structures in sequence were identified. Results Facial recess and its relative structures appeared in the 8th13th layers below the arcuate eminence.The emergence rates of facial nerve,facial recess,chordal eminence and chorda tympain nerve were all 100 percent,while the rate of ponticulus’s was 70 percent.From top to bottom, the distance from facial nerve to chorda tympani nerve became larger,and the largest distance was in the round window niche layer: right side was (5.20±0.06)mm,and left side was(5.16±0.09) mm in male,while right side was (5.16±0.05)mm,left side was (5.10±0.08)mm in female.The differences of two sides were not statistically evident(P>0.05).Conclusion The thin section collodion anatomy of temporal bone area,combined with the images of HRCT can clearly d Related Articles | Metrics

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ENZYME LABELED FLUORESCENCE 97 TO DETECT THE ACTIVITY OF ALKALINE PHOSPHATASE IN NEUTROPHILS 2007, 38 (5):  625-627.  doi: Abstract ( )   Objective To localize endogenous alkaline phesphatase in human neutrophils with ELF (enzyme-labeled fluorescence)-97 phosphate, which yields an intensely fluorescent yellow-green precipitate at the site of enzymatic activity. Methods Neutrophils were isolated from human blood and fixed,then histochemistry with the ELF-97 phosphate was performed with the ELF-97 endogenous phosphatase detection kit.All photography was performed with a Nikon labophot fluorescence/DIC microscope. Results Fluorescent yellow-green granules, well-distributed but size-varied, were observed in neutrophils under fluorescence microscope.There were 94.102±3.133 percent rate of positive neutrophil alkaline phosphatase among 51 cases of healthy volunteers.Conclusion ELF-97 endogenous phosphatase detection kit can provide sensitive,high-resolution localization of endogenous phosphatase activity in neutrophils. Related Articles | Metrics

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A FAST, ECONOMICAL AND SIMPLIFIED METHOD TO CULTURE AND PURIFY SCHWANN CELLS IN VITRO 2007, 38 (5):  628-630.  doi: Abstract ( )   Objective To explore a fast, simplified and economical method for Schwann cells(SCs) cultured and purified in vitro.In this way, a stable and reliable cell sources can be provided in order to study SCs transplanted in vivo. Methods SCs cultures were prepared from the sciatic and brachial nerves of 3 to 5-day-old SD neonatal rats with double enzyme digestion method to acquire dissociated cells and one enzyme digestion method to acquire incomplete digested tissues. Results With the same number of neonatal rats,one enzyme digestion(hemi-explant)method acquired at least two times more SCs than double enzyme digestion(single cell) method did and saved at least 40 minutes.96% of S-100 positive SCs cultured were shown by immunohistochemical staining.Conclusion When the one enzyme digestion method(hemi-explant) is used to culture SCs, sufficient SCs with qualified purity can Related Articles | Metrics