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2008, Volume 39 Issue 6
06 December 2008 Previous Issue    Next Issue
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REPAIRING EFFECTS OF NERVE GROWTH GRANULE ON RAT COMMON PERONEAL NERVE TRANSECTION INJURY 2008, 39 (6):  783-789.  doi: Abstract ( )   Objective To study the repairing effects of nerve growth granule (NGG) on rat common peroneal nerve transection injury. Methods After 50 Sprague-Dawley rats were subjected to nerve suture after transaction, they were randomly divided into 5 groups for daily intragastric administration of drugs: NGG high-dose (5.2g/kg), medium-dose (2.6g/kg), low-dose (1.3g/kg) groups, mecobalamin group (positive control) at 625 μg/kg, control group (control group control). The drug administration lasted for 4 weeks. Footprint test was performed 2-, 3- and 4- weeks after surgery to evaluate toe spread function (TSF). Electrophysiology was performed 4 weeks after operation to determine the compound muscle action potential (CMAP) and nerve action potential (NAP). The number of regenerated myelinated nerve fibers, thickness of myelin sheath and cross sectional area of tibial muscle were measured by histomorphology. Results TSF, amplitude and recovery rate of CMAP and NAP, the number of regenerated myelinated nerve fibers, thickness of myelin sheath and section area of tibial muscle were all increased significantly in a dose-dependent manner compared with the control group. Conclusion NGG contributes to axon growth and myelination, and thus promotes peripheral nerve regeneration in rats with functional recovery. Related Articles | Metrics

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THE FUNCTION OF AMNION IN TNE DIFFERENTIATION OF HUMAN UMBILICAL CORD BLOOD STEM CELLS INTO DOPAMINERGIC NEURONS 2008, 39 (6):  790-794.  doi: Abstract ( )   Objective To investigate the function of amnion endothelial cell in the differentiation of human umbilical cord blood stem cells into dopaminergic neurons. Methods Primary human amnion endothelial cells were separated and cultured in vitro; the conditioned medium(CM) was prepared through high speed centrifugation. The cord blood mesenchymal stem cells of PSUB>1/SUB> passage were induced by the conditioned medium, and the mophology of cells was observed under the inverted phase contrast microscope. The expression of tyrosine hydroxylase（TH）and dopamine transportor (DAT) of the induced cord blood mesenchymal stem cells were detected by immunocytochemistry staining method and immunoblotting(Western blotting). Results The masculine rate of TH and DAT of the cord blood mesenchymal stem cells of PSUB>1/SUB> passage which were induced by amnion endothelial cells conditioned medium was higher than that of the control group, with a significant difference (EM>P/EM>0.01 ). The results of Western blotting were in agreemetn with the immunocytochemistry staining. Conclusion Amnion endothelial cells can induce human umbilical cord blood stem cel Related Articles | Metrics

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EFFECTS OF INTRANIGRAL INJECTION OF GLUTAMATE AND GABA ON THE EXPRESSION OF DIVALENT METAL TRANSPORTER 1 AND HEPHAESTIN IN THE CAUDATE PUTAMEN NUCLEUS OF RATS 2008, 39 (6):  795-799.  doi: Abstract ( )   Objective To investigate the effects of intranigral injection of glutamate and gama amino butyric acid (GABA) on iron metabolism in the caudate putamen nucleus of rats. Methods Glutamate and GABA respectively were stereotaxically injected into the substantia nigra of rats. The levels of iron in the caudate putamen nucleus were detected by spectrophotometer. The levels of tyrosine hydroxylase (TH) in the substantia nigra, divalent metal transporter 1 without iron response element in the 3′-UTR (DMT1-IRE) and hephaestin (HP) in the caudate putamen nucleus were also detected by immunohistological study. Results The levels of iron in the caudate putamen nucleus of the mono sodium glutamate (MSG) group increased dramatically, while the levels of iron in GABA group decreased with no obvious change. Both the expression of TH in the MSG group and GABA group showed no differences when compared to controls. The levels of DMT1-IRE in the caudate putamen nucleus of the MSG group increased dramatically, while the levels of DMT1-IRE in the GABA group decreased dramatically. However, the levels of HP in the caudate putamen nucleus of MSG group decreased dramatically, while the levels of HP in GABA group increased dramatically. Conclusion Glutamate and GABA in the substantia nigra may affect iron metabolism through DMT1-IRE and HP in the caudate putamen nucleus. Related Articles | Metrics

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EFFECTS OF CHRONIC MORPHINE DEPENDENCE AND WITHDRAWAL ON pCREB PROTEIN EXPRESSION IN THE DISTAL CEREBROSPINAL FLUID CONTACTING NEURONS 2008, 39 (6):  800-803.  doi: Abstract ( )   Objective To investigate the changes in pCREB protein expression in the distal cerebrospinal fluid contacting neurons induced by chronic morphine dependence and withdrawal. Methods Twenty-four SD rats of both sexes were randomly divided into 3 groups (EM>n/EM>=8):the control group;the chronic morphine dependence group;the chronic morphine abstinence group. The methods of intraventricular injection of CB-HRP and CB-HRP/pCREB double-labeling in immunohistochemistry methods were used to observe the expression of pCREB in the distal cerebrospinal fluid contacting neurons. Results The CREB protein was activated in the distal cerebrospinal fluid contacting neurons induced by chronic morphine dependence and withdrawal, the numbers of CB-HRP double-labeling neurons in the two experimental groups were significantly elevated 223.0±61.5 and 213.0±50.6 respectively； when compared to the control expression of pCREB protein (P<0.01). Chronic morphine dependence and withdrawal showed no differences on the expression of pCREB protein (EM>P/EM>>0.05).Conclusion The distal cerebrospinal fluid contacting neurons were involved in the formation of chronic morphine dependence and withdrawal by activating the CR Related Articles | Metrics

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EFFECT OF INFLAMMATORY RESPONSES IN MICROGLIA INDUCED BY OLIGOMERIC β-AMYLOIDSUB>1-42/SUB>ON NEURONAL CELLS 2008, 39 (6):  804-809.  doi: Abstract ( )   Objective To investigate the effect and possible mechanisms of Oligomeric β-amyloidSUB>1-42/SUB>-induced inflammatory responses in microglia on neuronal cells. Methods BV-2 microglial and Neuro-2A neuronal-like cells were cultured EM>in vitro/EM>. Conditioned media from Oligo-AβSUB>1-42/SUB> stimulated BV-2 cells (Aβ-CM) was performed. Neuro-2A cells viability were measured by MTT assay. And the percentage of apoptotic or necrotic neuronal cells were evaluated by epifluorescence microscopy after stained with AO-EB. Tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and prostaglandin E2 (PGESUB>2/SUB>) le BR>vels were determined by ELISA assay. Nitric oxide (NO) levels were detected by Greiss assay. The levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins were detected by Western blotting. Results Treatment of neuronal cells with conditioned media from Oligo-AβSUB>1-42/SUB>-stimulated in BV-2 cells (Aβ-CM) resulted in a more dramatic decrease in the viability of Neuro-2A cells when compared to the Oligo-AβSUB>1-42/SUB> alone group when the concentration of induced Oligo-AβSUB>1-42/SUB> was at 1.0 and 5.0μmol/L (P<0.05,P<0.01). The combined groups(Aβ-CM+Aβ) showed significant interaction between Aβ-CM and Aβ(1.0μmol/L) by two-way ANOVA. Further, the percentage of apoptotic and necrotic neuronal cells was increased gradually and significantly with increasing concentrations of Aβ in CM by AO-EB stain assay. Me Related Articles | Metrics

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EFFECTS OF IGFBP-3/RXRα OR STAT-1 SIGNALING PATHWAY ON AβSUB>1-42/SUB> INDUCED APOPTOSIS IN RAT HIPPOCAMPUS NEURONS 2008, 39 (6):  810-815.  doi: Abstract ( )   Objective To investigate the effects of IGFBP-3, RXRα and STAT-1 on AβSUB>1-42/SUB> induced apoptosis in rat hippocampus neurons. Methods Apoptosis was induced by fibrillar AβSUB>1-42/SUB>. The percentage of neurons apoptosis was evaluated by microscopy after staining with TUNEL/DAPI. IGFBP-3 and RXRα positive neurons were observed by immunofluorescence. The expression of RXRα and STAT-1 protein were detected by Western blotting. Results After treatment with 20μmol/L AβSUB>1-42/SUB> for 24 hours, the apoptotic hippocampus neurons were shown by TUNEL/DAPI assay. The percentage of apoptotic neurons was increased in a time-dependent manner. During the development of apoptosis, both the percentage of IGFBP-3/ RXRα positive neurons and the expression of RXRα protein increased markedly after 3-6hours (EM>P/EM><0.01). However, the expression of STAT-1 protein significantly dec Related Articles | Metrics

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TYROSINE PROTEIN KINASE-DEPENDENT LONG-TERM POTENTIATION IN RAT BASOLATERAL AMYGDALA 2008, 39 (6):  816-819.  doi: Abstract ( )   Objective To compare the effects of different intervals of theta burst stimulation (TBS) on the expression of long-term potentiation (LTP) and to explore whether LTP is tyrosine protein kinase (TPK) -dependent.in basolateral amygdala (BLA). Methods Basolateral amygdala slices of rats were prepared. Field excitatory post-synaptic potentials (field potential, fEPSPs) of BLA were recorded by stimulating the external capsule. Two TBS’s were applied to induce LTP in BLA. Each TBS included a brief, high-frequency pulse train (5 pulses at 100 Hz) given at the theta-rhythm (5Hz) for 4 seconds. Experiments compare the effects of different intervals of two TBS’s on the expression of LTP in BLA. The role of tyrosine protein kinase (TPK) on LTP was then determined using bath application of TPK inhibitor genistein. Results Two TBS’s of 10 seconds interval failed to induce LTP in BLA. However, two TBS’s increased to 10 min and 30 min intervals, individually, both types of stimulations enhanced f-EPSPs. The enhanced f-EPSPs lasted for more than 30 min. LTP induced by two TBS’s of 10 min and 30 min interval were blocked by the TPK inhibitor genistein. Conclusion Two TBS’s of 10 min intervals was better at the induction of LTP in BLA. The activation of TPK was possibly Related Articles | Metrics

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EFFECT OF CHRONIC MULTIPLE STRESS ON THE EXPRESSION OF GROWTH-ARREST-SPECIFIC PROTEIN 7 IN THE HIPPOCAMPUS OF RATS 2008, 39 (6):  826-830.  doi: Abstract ( )   Objective To study the expression of growth-arrest-specific protein 7 (Gas7) in the hippocampus of rats after chronic multiple-stress. Methods thirty-six Wistar rats were randomly divided into two groups: the chronic multiple stressed group and the control group. Rats in the stressed group were administered with four kinds of stressors including vertical rotation，sleeping deprivation，restraint(6hours/day) and illumination irregularly and alternately for 6 weeks. Then the expression of Gas7 protein in the hippocampus of rats was assayed by immunocytochemical method and Western blotting technique respectively and apoptotic cells were observed. Results The expression of Gas7 was extensively found in the hippocampus of both the control group and the experimental group mainly in the endochylema and processes of the neurons in the CA1 area. Positive staining, average absorbance in the CA1 and dentate gyrus of the stressed rats were higher than that in the control group (P<0.05). There was no difference in the expressions in the CA3 areas in the two groups (P>0.05). Caspase-3 positive cells were also obserued in the stressed rats. Western blotting detection indicated that the expression of Gas7 in the stressed rats was significantly higher than that of the control group(P<0.05).Conclusion Gas7 may participate in protecting neurons in the stess and promote the neurogenesis an development of neurons. Related Articles | Metrics

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BIOCOMPATIBILITY OF SILK FIBROIN WITH BONE MARROW STROMAL CELLS OF MICE EM>IN VITRO/EM> 2008, 39 (6):  831-835.  doi: Abstract ( )   Objective To investigate the biocompatibility of bone marrow stromal cells (BMSCs) of mice EM>in vitro/EM> with silk fibroin materials and to explore a novel scaffold material to fabricate tissue-engineered nerve with introduction of BMSCs. Methods BMSCs were typically isolated from other cells by adherence to plastic. The mice-derived bone marrow stromal cells were cultured on the substrate of silk fibroin fibers and the cell attachment and growth during culture was observed by using light and electron microscopy. Mice-derived BMSCs were also cultured in the silk fibroin extract fluid. The cell ultrastructure was observed by transmission electron microscopy. MTT test was used to detect cell viability in different media for 12, 24, 48, 72 hours and 7 days respectively (the test was repeated 12 times for each group). Flow cytometry was employed to detect BMSCs cell cycle and phenotypes (the test was repeated 3 times). Results BMSCs cells were tightly attached to the silk fibroin fibers and grew along the silk fibroin fibers; some of them enwrapped the silk fibroin fibers and they exhibited either a spherical or spindle shape. The results of transmission electron microscopy, MTT test and flow cytometry analysis showed that there was no significant difference between BMSCs cultured in the silk fibroin extract fluid and those in plain IMDM medium in their morphology, cell viability, proliferation and phenotypes.Conclusion These data indicate that silk fibroin has good biocompatibility with BMSCs and is also beneficial to the survival of BMSCs without exerting any significant cytotoxic effects on their phenotype; thus it’s a potential scaffold material to fabricate tissue-engineered nerve with introductio Related Articles | Metrics

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ECTOGENIC TELOMERASE CATALYTIC SUBUNIT DOES NOT INFLUENCE THE CHARACTERISTIC OF HUMAN BONE MESENCHYMAL STEM CELLS 2008, 39 (6):  836-840.  doi: Abstract ( )   P>Objective To study the biological characteristics of human bone mesenchymal stem cells(hBMSCs) which were expressed telomerase catalytic subunit. Methods HBMSCs cells were infected by the retroviral particles（pBabepuro- hTERT）, and then the infected hBMSCs were selected by puromycin.We named infected HBMSCs as hTERT-hBMSCs.The telomerase activity of hTERT-hBMSCs and HBMSCs were detected by TRAP (PCR)-ELISA；The life character of hTERT-hBMSCs were observed by MTT, chromatosome kayotype，the rate of colony formation and osteoblast differentiation. Results TRAP (PCR)-ELISA detected that the telomerase activity of hTERT-hBMSCs was positive. The phenotype of hTERT-hBMSCs was CD44 positive and CD34 negative.The growth speed of hTERT- hBMSCs was higher than that of hBMSCs.Presently it had propagated eleven generations, but didn’t exhibit senescent phenomenon.hTERT-hBMSCs showed that chromatosome was nomal diploid，not aberration，the rate of colony formation was lower and the differentiational osteoblast capability of stem cells were maintained.Conclusion Infected Exogenic hTERT in Related Articles | Metrics

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EXPRESSIONS OF p53/Rb CELLULAR TRANSDUCTION PATHWAY RELATED GENES AND PROTEINS IN AGING RATS TESTES 2008, 39 (6):  841-844.  doi: Abstract ( )   Objective To investigate the significance of p53/Rb cellular transduction pathway related genes and proteins in testis aging by detecting expressions of p53, Rb, p16, p19 and p21 in aging rat testis. Methods Thirty rats of 8-week-old male SD rats (weight: 180-220g) were randomly divided into 2 groups, 15 rats in each: the normal control group and the aging model group. The sub-acutely aging model rats were made by injecting D-gal into abdominal cavity continually. RT-PCR was used to detect expressions of p53 and Rb genes in the testes of rats and Western blotting to detect the expressions of phosphorylated Rb, p16, p19 and p21 proteins in the testes of rats. Results The expression of p53 and Rb in the testes of model rats were obviously higher than those in the normal control group (P0.01 ); The expressions of p16, p19 and p21 proteins increased obviously in the testes of model group rats compared with the normal group, while the phosphorylated Rb protein expression decreased significantly (P0.01 ). Conclusion The expressions of p53/Rb cellular transduction pathway related genes and proteins changed obviously in aging rat testes, which suggests that the blocking of p53/Rb cellular transduction pathway may play a certain role in the aging of testis. Related Articles | Metrics

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PROTEIN MICROARRAY ANALYSIS OF EFFECTIVE INGREDIENT THAT BONE MARROW STROMAL CELLS CONDITIONED MEDIUM REGULATE THE DIFFERENTIATION OF NEURAL STEM CELLS 2008, 39 (6):  845-849.  doi: Abstract ( )   Objective To detect the cytokines in the bone marrow stromal cells (BMSCs) conditioned medium (neurobasal conditioned medium ,N-CM) that can regulate the differentiation of neural stem cells (NSCs) into high proportional neurons and explore the mechanism that BMSCs regulate the differentiation of NSCs.Methods The collected N-CM was divided into two parts(>5kD and <5kD)by means of ultrafiltration after misce bene. The two parts were used to culture NSCs separately, and the effective part that could regulate the differentiation of NSCs was detected by protein microarray analysis. Results The N-CM>5kD could promote the NSCs to differentiate into high proportional neurons, so this part was detected by protein microarray analysis, 7 cytokines CINC-3, CNTF, IFN-γ, IL-1α, MCP-1, TIMP-1and VEGF were detected to up-regulate 1.5 times compared with the neurobasal medium (molecular weight above 5kD). Conclusion The dissolvable molecules excreted by BMSCs could promote the NSCs to differentiate into neurons. Some cytokines with the molecular weight of above 5kD play a crucial role during this process. Related Articles | Metrics

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EFFECTS OF HYPOXIA ON THE EXPRESSIONS OF HIF SUBUNITS HIF-1α,HIF-2α AND HIF-β IN HUMAN LUNG CANCER CELL LINES AND THEIR CORRELATIONS 2008, 39 (6):  850-853.  doi: Abstract ( )   Objective To explore the effects of hypoxia on the expressions of hypoxia inducible factor-1α（HIF-1α）, hypoxia inducible factor-2α (HIF-2α) and hypoxia inducible factor-β (HIF-β) in human lung cancer cell lines and their correlations. Methods Six lung cancer cell lines (SPCA1，A549，H446，SH77，H520 and 95D) were incubated under 0.5% OSUB>2/SUB> for 4 to 24 hours.Western blotting and real-time RT-PCR were used for protein and mRNA analysis of HIF-1α, HIF-2α and HIF-β. Results Both HIF-1α and HIF-2α mRNAs were at very low levels or almost undectable under normoxia, as well as their proteins. HIF-1α mRNA experienced a decrease after transcient hypoxia, while the level of HIF-2α mRNA increased. By contrast, under chronic hypoxia, the levels of both HIF-1α and HIF-2α mRNA increased steadily. HIF-1α and HIF-2α proteins were accumulated and changed differently under prolonged hypoxia. HIF-1α was expressed in all the cell lines at various levels, but HIF-2α protein was absent in some cell lines. While both the levels of HIF-β mRNA and HIF-β protein w Related Articles | Metrics

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EFFECT OF HYPOXIA PRECONDITIONING ON THE MITOCHONDRIA ULTRASTRUCTURE OF HEPATOCYTE DURING LIVER TRANSPLANTATION IN RATS 2008, 39 (6):  854-857.  doi: Abstract ( )   Objective To observe the effects of hypoxia preconditioning on the mitochondria ultrastructure of hepatocyte during liver transplantation in rats. Methods A modified orthopotic liver autotransplantation model was used to simulate liver transplantatation. Seventy two Sprague-Dawley rats were randomly divided into the following three groups:the normal control (group NC), the autotransplantation group (group AT), and the hypoxia preconditioning group (group HP), with twenty four rats in each group. Group HP were given an 8% oxygen atmosphere for 90 minutes before the operation. At 1, 6, and 24 hours after the operation, the rats were killed and the following tests were conducted: 1. The morphology of mitochondria was observed under transmission electron microscopy (TEM); 2.Mitochondria were quantitatively analyzed by a MiVnt image analysis system. Results Hepatic cells in group AT showed typical injury signs under TEM, the appearance of hepatocytes and mitochondria in group HP were much better than the group AT, and the areas, perimeters and diameters of mitochondria in group HP were much smaller than those in group AT by a significantly amount (EM>P/EM><0.05). Conclusion Changes of mitochondria ultrastructure were the early events in hepatocellular damage, which could be relieved by HP. Related Articles | Metrics

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EXPRESSIONS OF TRANSFORMING GROWTH FACTOR-BETA 1 AND ITS TYPE Ⅱ RECEPTOR IN EXPERIMENTAL RAT ASCENDING AORTIC ANEURYSM 2008, 39 (6):  858-861.  doi: Abstract ( )   Objective To investigate the expressions and significance of transforming growth factor-beta 1(TGFβ1) and its typeⅡ receptor (TGFβRⅡ) in experimental rat ascending aortic aneurysm of rat model. Methods The rat ascending aortic aneurysm models were made by banding ascending aorta of young Wistar rats. The ascending aortas were taken 4 months after operation. Immunohistochemistry staining and Western blotting were used to investigate the expressions of TGFβ1 and TGFβRⅡ.Result Immunohistochemistry staining results showed that TGFβ1 expressed in all layers of the aortic aneurysm and the control. TGFβRⅡ was extensively located in the hyperplastic intima and tunica media smooth muscle cells in the aortic aneurysm, while there was only a little positive staining in the control group. Western blotting results indicated that the expression levels of TGFβ1 and TGFβRⅡ in the aortic aneurysm were stronger than the co Related Articles | Metrics

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EXPRESSIONS OF NUCLEOSTEMIN， EGF AND EGFR mRNA IN HUMAN ESOPHAGEAL SQUAMOUS CELL CARCINOMA TISSUE 2008, 39 (6):  862-866.  doi: Abstract ( )   Objective To investigate the expressions of nucleostemin(NS), epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR) mRNA and their relationship in human esophageal squamous cell carcinoma(ESC) tissue. Methods The expressions of NS, EGF and EGFR mRNA in 62 cases of esophageal squamous cell carcinoma, 31 cases of dysplasia and 62 cases of normal esophageal tissue were determined by EM>in situ/EM> hybridization. The relationship of NS, EGF and EGFR mRNA with the tumor grading, the infiltration and lymphatic metastasis of ESC was analyzed. Results In the normal esophageal tissue, dysplasia esophageal tissue and ESC tissue, the positive expression rates of NS mRNA were 21.0% (13/62), 25.8% (8/31) and 69.4% (43/62) respectively; the positive expression rates of EGF mRNA were 40.3% (25/62), 48.4% (15/31) and 77.4% (48/62); the positive expression rates of EGFR mRNA were 30.6% (19/62), 45.2% (14/31) and 75.8% (47/62). The expressions of NS, EGF and EGFR mRNA showed higher positive correlation with the tumor grading, the infiltration and lymphatic metastasis of ESC (all P<0.05). The expression of NS mRNA was positively correlated with that of EGF and EGFR mRNA (EM>r/EM>=0.394 and EM>r/EM>=0.604, EM>P/EM><0.05)in ESC. Conclusion The NS, EGF and EGFR mRNA may play an important role in the genesis and progression of ESC. Related Articles | Metrics

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INHIBITION OF VEGF-C GENE BY siRNA EXPRESSION VECTOR IN HUMAN BREAST CANCER CELL LINE MCF-7 2008, 39 (6):  867-871.  doi: Abstract ( )   Objective To construct small interfering RNA（siRNA）expression vector and to investigate its inhibitory effect on vascular endothelial growth factor-C（VEGF-C） expression in human breast cancer cell line MCF-7 Methods A pair of hairpin-like oligonucleotide sequences specific for VEGF-C gene were designed and synthesized.The annealed oligonucleotide fragments were subcloned into plasmid pRNAT-U6.1/Neo to construct VEGF-C siRNA expressing vector.Then VEGF-C siRNA expression vectors were confirmed by PCR and sequencing.Transfection of VEGF-C siRNA expressing vector into MCF-7 cells was performed using liposome transfection reagents.VEGF-C mRNA and protein expression were examined by RT-PCR and immunocytochemistry respectively. Results PCR and DNA sequencing showed that siRNA expression vector targeting VEGF-C was successfully constructed.VEGF-C expression in the cells transfected by VEGF-C siRNA expression vectors was inhibited significantly at both mRNA and protein levels.VEGF-C mRNA and protein expression were decreased 61.8% and 78.3% compared with control plasmid respectively（EM>P/EM>0.05）.Conclusion VEGF-C siRNA expression vector can effectively inhibit the expression of VE Related Articles | Metrics

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EFFECTS OF LOW-DOSE MIFEPRISTONE ON HUMAN GRANULOSA CELLS 2008, 39 (6):  872-876.  doi: Abstract ( )   Objective To explore the effects of low-dose mifepristone on human granulosa cells and the molecular mechanism of its inhibiting or delaying ovulation. Methods Phase contrast microscopy and electron microscopy were conducted to observe the morphological and biochemical change of granulosa cells exposed to various doses of mifepristone for 24 hours (the final concentrations of mifepristone were respectively 2.5μmol/L, 5μmol/L and 10μmol/L, Non-mifepristone as the control group). Immunocytochemistry was performed to detect the protein expressions of Bcl-2 and Bax in granulosa cells exposed to various doses of mifepristone for 24 hours. The mRNA levels of Bcl-2 and Bax in granulosa cells were detected by RT-PCR. Results Granulosa cells treated with various doses of mifepristone presented typical morphological characteristics of apoptosis such as condensation of chromatin and cytosolic, and shrink of cells. The expression of Bcl-2 protein might be down-regulated, while the expression of Bax might be up-regulated by mifepristone. The histological score (HSCORE) of Bcl-2 protein in granulosa cells, which were incubated with various doses of mifepristone (2.5μmol/L, 5μmol/L, 10μmol/L) for 24 hours, were respectively 2.15±0.16,1.88±0.13 and 1.64±0.16 respectively，Compared with the HSCORE of the control group (2.51±0.16), the difference was significant （EM>P/EM><0.001）. Compared with the control group (1.50±0.16), there was singificant difference in the expression of Bax protein (1.85±0.10, 2.00±0.18 and 2.29±0.20) （EM>P/EM> Related Articles | Metrics

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EFFECTS OF HYDROGEN SULFIDE ON INTRACELLULAR CALCIUM AND PROLIFERATION IN HEPATIC STELLATE CELLS EM>IN VITRO/EM> 2008, 39 (6):  877-880.  doi: Abstract ( )   Objective To study the effect of hydrogen sulfide (HSUB>2/SUB>S) on intracellular calcium concentration and proliferation in hepatic stellate cells (HSC) and the possible mechanism of HSUB>2/SUB>S. Methods HSC-T6, an activated rat HSC, was plated in 35-mm culture dish at a density of 1×10SUP>5/SUP>/ml, and rotuinely incubated in DMEM media for 24 hours. The intracellular CaSUP>2+/SUP> loaded by Flu-3/AM was measured by laser scanning confocal microscope. The dynamic changes of intracellular CaSUP>2+/SUP> fluorescence intensity(［CaSUP>2+/SUP>］i ) stimulated by NaSH, a HSUB>2/SUB>S donor, were measured. The HSC proliferations were measured by MTT at different HSUB>2/SUB>S concentrations. Results Low concentration of HSUB>2/SUB>S(100μmol/L) decreased significantly HSC-T6 ［CaSUP>2+/SUP>］i (16.20±3.56 vs 14.12±3.76,EM>P/EM><0.05). The increased HSC-T6 proliferations were also observed in low concentration stimulation of HSUB>2/SUB>S(50-100μmol/L). The decreased ［CaSUP>2+/SUP>］i was inhibited by glibenclamide, a KSUB>ATP/SUB> channel antagonist， significantly when administered with HSUB>2/SUB>S of higher concentrations. However, high concentration of HSUB>2/SUB>S(1mmol/L) increased significantly HSC-T6 ［CaSUP>2+/SUP>］i，P<0.05, with no significant changes in proliferation. Conclusion Th Related Articles | Metrics

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IMMUNOFLUORESCENCE ANALYSIS OF MANCHETTE DURING MOUSE SPERMIOGENESIS 2008, 39 (6):  881-885.  doi: Abstract ( )   Objective To investigate the localization and the morphological changes of manchette during mouse spermiogenesis. Methods Immunofluorescence staining with FITC and costaining with DAPI were used to demonstrate the cellular localization of the manchette at different stages during mouse spermiogenesis. The structural changes of the manchette were observed during the maturing of the spermatid. Results Immunofluorescence staining showed that manchette existed exactly around the nuclei of the spermatids. Manchette began to form, when the shape of the nucleus changed from spherical to slightly elongated. While the nucleus of the spermatids condensed and elongated at later stages, manchette moved gradually to the caudal position of the spermatids. At last, the manchette diminished as the spermatids became mature. During mouse spermiogenesis，manchette underwent a transition from a cap-like to a tubular configuration. ConclusionThe formation and diminishment of the manchette is in step with the condensation and elongation of the nucleus of the spermatid. Both the structural and positional changes of the manchette coincide with the changes of the nucleus. These results imply that manchette might play an important role in mouse spermiogenesis. Related Articles | Metrics

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EXPRESSION OF FOXC2 DURING LYMPHANGIOGENESIS IN HUMAN EMBRYO 2008, 39 (6):  886-889.  doi: Abstract ( )   Objective To study the human lymphangiogenesis in the early stage of embryo and the expression of forkhead box C2(FOXC2) in human lymphangiogenesis. Methods Lymphatic vessel endothelial haluronic acid receptor-1(LYVE-1) was used as the special marker of lymphatic vessels.Immunohistochemical and double immunofluorescence staining were used to detect the expressions of FOXC2 and LYVE-1 in lymphatic vessels of 85 human embryos of 5 to 11 weeks pregnancy and analyze the features of lymphatic vessel genesis and development. Results LYVE-1 was initially detected in lymphatic vessels in 7-week human embryos. The lymphatic vessel endothelial cells presented brown staining for LYVE-1 in jugular and thoracic region. LYVE-1 was also found to express in human embryonic mesentery lymphatic vessels on the 70th day of pregnancy. The expression of FOXC2 in human embryo was detected prior to that of LYVE-1 At the beginning of the 6th week of pregnancy, FOXC2 was obviously seen in human embryonic mesoderm mesenchyme.FOXC2 expressed not only in human embryonic lymphatic vessels,but also in the vertebral body,cardiovascular wall and other tissues.The expressions of FOXC2 and LYVE-1 were still seen in human embryonic lymphatic vessel endothelial cells after 80 days of pregnancy. Conclusion Lymphangiogenesis of human embryo begins between the 7th and 8th weeks of pregnancy. FOXC2 and LYVE-1 could have some relation with the human embryonic lymphatic vessel genesis and development. Related Articles | Metrics

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FGFR3 EXPRESSION PATTERN DURING THE DEVELOPMENT OF MOUSE EMBRYONAL LIMB BUD DETECTED BY MICROARRAY ANALYSIS 2008, 39 (6):  890-893.  doi: Abstract ( )   Objective To study the expression pattern of fibroblast growth factor receptor 3(FGFR3) in mouse embryonal limb bud development. Methods Affymetrix mouse 430 2.0 cDNA was used with microarray technique to examine the 34 000 genes. Histology staining observation was performed at the same time. Results FGFR3 expression was enhanced obviously in mouse E12.5d embryonal limb bud. It continued to increase and peaked in mouse E13.5d embryonal limb bud. Furthermore, the expression down-regulated gradually from E14.5d in mouse embryonal limb bud. Conclusion The expression of FGFR3 has close correlation with chondrogenesis of mouse embryonal limb bud. FGFR3 may play an important role in chondrogenesis. Related Articles | Metrics

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CHANGES OF SOMATOSTATIN RELATED SIGNAL TRANSDUCTION MOLECULARS IN RAT GASTRIC MUCOSA OF SPLEEN DEFICIENCY SYNDROME COMBINED WITH GASTRIC ULCER 2008, 39 (6):  894-900.  doi: Abstract ( )   Objective To investigate the role of somatostatin and related signal transduction molecules in the process of spleen deficiency syndrome combined with gastric ulcer. Methods Seventy adult male and female Wistar rats were selected to set up an animal model of spleen deficiency syndrome combined with gastric ulcer. Then immunohistochemistry, reverse-transcript PCR, Western blotting and hematoxylin-eosin staining were used to detect the changes of somatostatin, somatostatin receptor 2 mRNA, excellular signal_regulated kinase 2(ERK2) and histological structure of gastric mucosa respectively. Results In the gastric mucosa of spleen deficiency syndrome combined with gastric ulcer, the content of somatostatin increased and the mRNA of somatostatin receptor 2 increased, while the content of ERK2 decreased. Conclusion Somatostatin signal transduction pathway may probably play an important role in the onset of spleen deficiency combined with gastric ulcer. Related Articles | Metrics

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EFFECT OF PINEALECTOMY AND MELATONIN ON IL-7 EXPRESSION OF RAT THYMIC EPITHELIAL CELLS 2008, 39 (6):  901-905.  doi: Abstract ( )   Objective To study the effect of pinealectomy and melatonin on interleukin 7(IL-7) expression of rat thymic epithelial cells (TECs) and its physiological significance. Methods Cultured rat TECs were immunostained with anti-pan-cytokeratin; MTT was applied to investigate the growth of cultured rat TECs with melatonin treatment (1×10SUP>-3/SUP>-10SUP>-9/SUP>mol/L), semiquantitative RT-PCR was applied to assess IL-7 mRNA expression. Totally 110 clean_grade male SD rats were divided into five groups: normal control group, sham operation group, pinealectomized group, pinealectomy+75mg/(kgI>&#/I>8226;d) MLT group, pinealectomy +15mg/(kgI>&#/I>8226;d) MLT group, and took the thymus out in the 4th week and the 8th week, Immunohistochemistry and semiquantitative RT-PCR were applied to observe IL-7 expression in rat TECs. Results Melatonin made no difference on rat TECs growth, but IL-7 mRNA expression in TECs was increased by melatonin treatment in a dose dependent manner; Pinealectomy made no difference on the synthesizing of IL-7 by rat TEC, but 15mg/(kgI>&#/I>8226;d) melatonin treatment resulted in significant increasing of IL-7 expression in certain period. Conclusion pinealgland and melatonin may regulate the differentiation and development of thymocyte by IL-7 expres Related Articles | Metrics

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EFFECT OF TOTAL FLAVONOIDS OF EM>CHRYSANTHEMUM INDICUM/EM>ON THE APOPTOSIS OF ADJUVANT ARTHRITIS RAT SYNOVIOCYTESAND THE EXPRESSION OF CASPASE-3 2008, 39 (6):  906-909.  doi: Abstract ( )   Objective To study the effect of the extract of total flavonoids of Chrysanthemum indicum (TFC) on adjuvant arthritis(AA) synoviocytes. Methods Totally 0.1ml of the complete Freund’s adjuvant was subcutaneously injected into the right hind feet pads of 20 SD rats. 24 days after immunity synoviocytes in the knee joint were treated with TFC. Cell morphology was examined with electron microscopy. Protein level of caspase-3 cleaved fragments was analyzed by Western blotting. The annexin V stain assay was applied to explore the effect of caspase-3 inhibitor on the amelioration in synovial cells apoptosis of AA rats. Results Typical morphology and biochemical feature of apoptosis in synovial cells of AA rats were observed with TFC. The protein level of caspase-3 cleaved fragments increased obviously and was related with the concentration of TFC in synovial cells of AA rats. The apoptotic cells positively stained with annexin V were markedly reduced by caspase-3 inhibitor. Conclusion TFC can induce apoptosis in AA rats synoviocytes，which may achieve therapeutical effects in AA. The activation of caspase-3 may Related Articles | Metrics

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IMMUNOHISTOCHEMICAL ANALYSIS OF AGEs， TGF-β1 AND CTGF IN DIABETIC RAT LUNGS 2008, 39 (6):  910-914.  doi: Abstract ( )   Objective To approach the changes of advanced glycosylated endproducts (AGEs), transforming growth factor(TGF-β1) and connective tissue growth factors(CTGF) in the lungs of experimental diabetic rats and their relationship. Methods Totally 48 male SD rats were divided into the diabetes mellitus（DM） group and the control group，with 24 rats in each group.The DM rat model was made by the injection of streptozocin (60mg/kg) into the caudal vein. The rats were killed and the lungs were taken at the end of the 4 th, 12 th and 20th weeks respectively after the models were established. The changes of AGEs,CTGF,TGF-β1 in rat lungs were observed with immunohistochemical assay and the image was analyzed. Results A great quantity of AGEs positive cells were observed in the alveolar epithelial cells, bronchial mucosal epithelium, angio-endothelial cells and smooth muscle cells of the DM rats. The average gray(AG) was lower than that of the controls（EM>P/EM><0.05） and decreased with the DM course（EM>P/EM><0.01）. In the 4_week DM rats, there were a few CTGF and TGF-β1 positive cells in the bronchial mucosal epithelium, angio-endotheliial cells and lung interstitial cells. In the 12_and 20_week DM rats, there were a great many CTGF and TGF-β1 positive cells. The AG was lower than that of the controls（EM>P/EM Related Articles | Metrics

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PROPERTIES OF HPP-CFC OF CD133SUP>+/SUP> CELLS DERIVED FROM HUMAN PLACENTA 2008, 39 (6):  915-918.  doi: Abstract ( )   Objective To investigate whether human placenta tissue (PT) CD133SUP>+/SUP> cells possess the high proliferative potential colony forming cells(HPP-CFC) and analyze its properties in order to verify that human PT contains primitive hematopoietic stem/progenitor cells(HSPC). Methods Single cell suspension liquid of human PT was prepared by mechanical method. After that the mononucler cells(MNC) from PT was separated by Histopaque-1007 agent. CD133SUP>+/SUP> cells contained in MNC were isolated and purified by magnetic activated cell sorting(MACS) method. After 28 days of HPP-CFC expansion culture of CD133SUP>+/SUP> cells, the frequency and morphology of HPP-CFC were counted and observed. Phenotypes of purified CD133SUP>+/SUP> cells and HPP-CFC were analyzed by flow cytometer(FCM). In parallel, human umbilical cord blood(UCB) samples underwent the same protocols for comparison. Results After 28 days of culture the expansion fold of PTSUP>-/SUP>CD133SUP>+/SUP> cells was 266, lower than that of UCB-CD133SUP>+/SUP> cells which was 362 (P<0.01). The number of HPP-CFC in CD133SUP>+/SUP> cells derived from human PT and UCB were (32.4±11.2)/5×10SUP>3/SUP> and (17.7±5.7)/5×10SUP>3/SUP> respectively, the number of the former obviously higher than that of the latter (P<0.01). FCM analysis results showed that, except for CD133SUP>+/SUP>CD34SUP>-/SUP> subset from UCB-CD133SUP>+/SUP> cells, the proportion of CD133SUP>+/SUP>CD34+, CD133SUP>-/SUP>CD34SUP>+/SUP> subsets both from PT- and UCB-CD133SUP>+ /SUP>cells and CD133SUP>+/SUP>CD34SUP>- /SUP>subset from PT-CD133SUP>+/SUP> cells were decreased through 4 weeks of culture BR>.Conclusion Human PT-CD133SUP>+ /SUP>cells can form HPP- Related Articles | Metrics

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EXPRESSION OF HEPATOCYTE NUCLEAR FACTOR 6 DURING THE PROCESS OF INTRAHEPATIC BILIARY DEVELOPMENT 2008, 39 (6):  919-922.  doi: Abstract ( )   Objective To investigate the function of hepatocyte nuclear factor 6（HNF 6） during the development of intrahepatic biliary. Methods The expression of HNF6 at different stages of intrahepatic bile ducts embryonic development and in the adult mouse liver were detected by RT-PCR and immunohistochemical staining. Results RT-PCR results showed that the expression of HNF6 was detected in the liver at E9d, the time of liver formation onset. Then, HNF6 disappeared transiently at E13d, but it appeared again at E15d. Its expression lasted till birth. According to the immunohistochemical staining studies, the immunohistochemical positive reaction of CK19, which dispersed in the liver cords, appeared at E13d. At E15d, CK19-positive cells formed a single cell layer called the ductal plate around the branches of the portal vein located close to the hilum. Later, these immunohistochemical positive cells were confined to the ductal plate and biliary epithelial cells. The immunohistochemical positive reaction of HNF6 was detectable in most cells of liver cords at E9-11d, which became undetectable at E13d. Similar to CK19 at E15-17d, the immunohistochemical positive reaction of HNF6 could be observed at P1d. BR>Conclusion HNF6 has no important role in the specification of liver during rat embryonic development. It may play a role in the onset of liver development, the differentiation of hepatic stem cell towards biliary epithelial cells and the maintenance of the morphological characteristic of biliary epithelial cells. BR> Related Articles | Metrics

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EXPRESSION OF ANNEXINA5 IN HUMAN UTERINE CERVICAL SQUAMOUS CELL CARCINOMAS 2008, 39 (6):  923-926.  doi: Abstract ( )   P>Objective To observe whether the expression of Annexin A5(ANXA5) changes in human uterine cervical squamous cell carcinomas(UCSCC). Methods 25 UCSCC tissues and 15 normal human uterine cervical tissues were collected. Each sample was lysed in lysis buffer.Whole protein of the supernatant was estimated by BCA-100 Protein Quantitative Analysis Kit. The expressions of ANXA5 in UCSCCs and normal uterine cervical tissues were detected respectively with Western blotting. To further ensure the expression of ANXA5 in UCSCCs, another 45 UCSCC specimens and 20 normal cervical tissues were collected. ANXA5 expression was detected by in situ hybridization and immunohistochemistry respectively. Results The staining of ANXA5 band with Western blotting in UCSCCs was much heavier than that in normal uterine cervical tissues and the expression of ANXA5 was found much higher in UCSCCs by immunohistochemistry and in situ hybridization.Conclusion Expression of ANXA5 was up-regulated in human UCSCCs.There’s some relationship between uterine cervical squamous cell carcinoma and ANXA5 p Related Articles | Metrics

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EFFECTS OF FOLLICLE-STIMULATING HORMONE ON THE SECRETION OF NGF AND EGF IN RAT SUBMAXILARY GLANDS EM>IN VITRO/EM> 2008, 39 (6):  927-930.  doi: Abstract ( )   P>Objective To investigate the distribution of nerve growth factor(NGF) and epidermal growth factor (EGF) and their colocalization with follicle-stimulating hormone (FSH) in rat submaxilary glands, and to study the effects of FSH on the secretion of NGF and EGF by submandibular gland tissue in vitro. Methods Immunohistochemical colocalization method was used in the experiment.Submandibular tissues of rats were incubated in vitro and different concentration of FSH was added.Enzyme link immunoassay was used to measure EGF and NGF in supernatant. Results The serous glandular cells, granular convoluted epithelial cells and all other duct epithelial cells in the submandibular gland showed FSH,NGF or EGF positive immunoreactivity.The positive substance was distributed in the cytoplasm with negative nuclei. Both FSH and NGF or EGF positive products were co-located in rat submandibular gland.When the concentration of FSH was more than 10SUP>-5/SUP>-10SUP>-6/SUP>IU/L,the secretion of EGF and NGF lessened with the decreased concentration of FSH; when the concentration of FSH was less than 10SUP>-5/SUP>-10SUP>-6/SUP>IU/L, the secretion of EGF and NGF increased with the decreased concentration of FSH. Conclusion FSH has the same bidirectional regulation effects on EGF and NGF EM>in vitro./EM> FSH may regulate the function of endocrine of rat submandibular gland. /P> Related Articles | Metrics

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STUDY OF BRONCHIAL ANATOMY OF LEFT LUNG WITH 64 SLICE SPIRAL CT 2008, 39 (6):  931-935.  doi: Abstract ( )   Objective To classify the segmental bronchial patterns of the left upper lobe by combining three post-processing images from 64 slice spiral CT and to study how to identify different ramifications in transverse thin-section CT. Methods Totally 204 patients with routine thorax scans were enrolled. The segmental bronchi were demonstrated in terms of bronchial tree, virtual bronchoscopy and thin-section CT three post-processing images. Integrated with the three post-processing images, the segmental bronchial patterns of the left upper lobar bronchi were classified into several main types, and displayed in transverse thin-section CT. Results The segmental bronchial ramifications of the left upper lobe were classified into three types mainly: common stem of apical and posterior segmental bronchi (64%, 130/200), trifurcation (23%，45/200), common stem of apical and anterior segmental bronchi (10%, 21/200), and they could be identified in two typical slices of transverse thin-section CT. There were two dominating types in the left basilar segmental bronchi: bifurcation (75%, 163/216), trifurcation(18%, 39/216), and they could also be identified in two typical slices of transverse thin-section CT. Conclusion The segmental bronchi of the left lung can be definitely classified by three post-processing images from 64 slice spiral CT. Related Articles | Metrics

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ANATOMIC STUDY OF CORONARY ARTERY SIZE USING 16-SLICE COMPUTED TOMOGRAPHY 2008, 39 (6):  936-940.  doi: Abstract ( )   Objective To evaluate the CT features of the coronary artery in normal adults and to measure the diameter of coronary artery using 16-slice computed tomography. Methods 16-slice CT coronary angiography was performed in this study. Totally 104 cases whose coronary arteries were normal were divided into 3 groups according to age and dominant pattern of coronary artery. The diameter of right artery (RCA), including the proximal, middle and distant segments were measured by using CT, so was the diameter of left main artery (LM), left anterior descending artery (LAD) including the proximal, middle and distant segments, and left circumflex artery proximal and distant segments. Results The diameters of coronary arteries increased with the age. There was a difference of LM diameter between the elder group and the youth group and middle group. However there was no difference in the youth group and the elder group. No difference were detected from other coronary artery segment’s diameter. The diameter of LM in left dominant pattern was the largest, and it showed a difference when compared with the other two groups, bt there was no difference between the latter two groups. The diameters of LAD and LCX in left dominant pattern were the largest, and that of in right dominant pattern were the smallest. The diameter of LAD had no difference among the three groups. The diameter of LCX proximal segment had no difference between the balanced dominant pattern and right dominant pattern, but there was a difference compare to the left dominant pattern. The diameters of LCX distant segment showed differences between any two groups. The diameter of RCA in the right dominant pattern was the largest. The diameters of RCA proximal segments showed no difference between the balanced dominant pattern and left dominant pattern, and there was difference among any other groups.Conclusion MSCT can display coronary artery from any angle and position. The diameter of coronary artery segment increases with the increase of age. Coronary artery size measured by MSCT can provide valuable information for coronary disease. Related Articles | Metrics

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EXPERIMENTAL STUDY ON THE CHANGES OF GELATINASE ACTIVITY OF ASCENDING AORTIC ANEURYSMS 2008, 39 (6):  941-943.  doi: Abstract ( )   Objective To investigate the activity changes of gelatinase in the formation of ascending aortic aneurysm. Methods Thirty five young Wistar rats were divided into two groups: the control group and the experiment group. The rat models induced by ascending aorta banding were made. The ascending aortas were taken after 3-5 months operation, changes of gelatinase activity was observed by gelatin zymography and film EM>in situ/EM> zymography. Results Gelatinase activity of ascending aortic aneurysm was significantly increased compared with that of normal ascending aortic aorta.Conclusion Elevation of gelatinase activity may play a significant role in the formation of ascending aortic aneurysm. Related Articles | Metrics

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THE EVALUATION OF FOCAL CEREBRAL ISCHEMICA REPERFUSION MODEL WITH MIDDLE CEREBRAL ARTERY OCCLUSION BY MICRO-BALLOON IN RHESUS MI>onKEY/I>S 2008, 39 (6):  944-947.  doi: Abstract ( )   Objective To establish an ideal focal cerebral ischemia reperfusion model in mI>onkey/I>s. Methods Adult healthy rhesus mI>onkey/I>s (Macaca mulatta) 12 cases (male 6 and female 6), aged 4-7 years and weighted 4.8-7.5kg. were used in this study. The middle cerebral artery occlusion/reperfusion (MCAO/R) model was established by inserting a standard micro-balloon catheter intraluminally from the carotid common artery or femoral artery into the proximal segment of the middle cerebral artery (MCA). The regional cerebral blood flow of MCA was occluded by expanding the micro-balloon to cause ischemia, and withdrawing the micro-ballon catheter to reperfuse the MCA. The MCAO/R model was evaluated by angiography, magnetic resonance angiography (MRA), magnetic resonance imaging(MRI), tetrazolium chloride (TTC) staining and neurological behavoral function scores. Results By inserting a micro-balloon catheter intraluminally from the carotid common artery or femoral artery into the MCA, the micro-balloon catheter could be inserted into the MCA to occlude blood flow, and no image of MCA shown on TV screen. In MCA blood flow supplied area, magnetic resonance T1, T2 and DWI showed high signals, TTC staining showed cerebral ischemic infarction, and correspondly the mI>onkey/I>s showed neurological function disorders. This method used a simple operatire procedure had a high successful rate, and could be repeated. Conclusion We showed ideal method to establish the MCAO/R model in mI>onkey/I>s by inserting intraluminally a micro-balloon catheter into the MCA. Related Articles | Metrics