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2007, Volume 38 Issue 6
06 December 2007 Previous Issue    Next Issue
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INFLUENCE OF OLFACTORY BULB REMOVAL ON THE PROLIFERATION AND DIFFERENTIATION OF NEW NEURONS ORIGINATED FROM THE SUBVENTRICULAR ZONE OF ADULT RAT BRAIN 2007, 38 (6):  631-635.  doi: Abstract ( )   Objective To explore the effect of olfactory bulb (OB) removal on neurogenesis in subventricular zone (SVZ) by observing the changes of proliferation and differentiation of new cells originated there. Methods An animal model of OB removal was created. Nissl staining and immunohistochemistry for showing the polysialylated form of the neural cell adhesion molecule (PSA-NCAM) and BrdU were done in the brain tissue from adult OB removal rats and observation was made under the light microscope.The total cell number and the number of PSA-NCAM and BrdUpositive cells on the two sides of RMS were counted and analyzed, and the morphological changes of PSA-NCAMpositive cells were observed. Results 1The total cell number and the BrdUpositive cell number on the right side of the RMS in the rats with OB removal increased compared with those on the left side, but the percentage of the BrdUpositive cells decreased apparently with the operation delay after OB removal. 2There were more PSANCAMpositive cells with long processes in the granular layer on the side of OB removal than those on the control side. In addition, some PSA-NCAMpositive cells with long processes were also observed in the RMS at the edge of the excision.Conclusion Young neurons still migrate along the RMS after the OB removal but the prolife Related Articles | Metrics

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THE EFFECT OF CORIARIA LECTONACTIVITED ASTROCYTECONDITIONED MEDIUM AND CNQX ON NSF OF THE CEREBRAL OF NORMAL RATS AND CULTURED NEURONS 2007, 38 (6):  636-641.  doi: Abstract ( )   Objective To explore the effects of astrocytes on the N-ethylmaleimide-sensitive fusion protin,(NSF) and AMPA receptor of the neurons as well as their function in epileptogenesis. Methods ACM was injected into lateral ventricle of SD rats and the behaviour changes were observed; Immunohistochemical method was used to assess the changes of the expression of NSF in the cerebral cortex and hippocampus; The cultured neurons were divided into control group, ACM group and CNQX+ACM group at random, immunocytochemistry was used to assess the changes of the expression of NSF, Western blotting was used to assess the changes of the content of NSF of the cultured neurons. Results Seizure was observed in ACM group 30 min after injecting ACM. the immunoreaction of NSF in hippocampus and cerebral cortex of rats were depressed than those of the control group 2h, 4h after injecting ACM (P<0.05); The immunoreaction of NSF in cultured neurons of the ACM group was weaker than those of the control group 4h, 8h after ACM (P<0.05). By mean of Western blotting, the concentrations of NSF of cultured neurons were depressed than those of the control group and the CNQX+ACM group 4h, 8h after ACM (P<0.05). Conclusion The results mentioned above indicated that the ACM can be reduced the expressions of NSF and result in epileptogenesis. CNQX can anatagonize this effect, showing that the downregulation of ACM on NSF of rats and cultured neurons was partly mediated by AMPA receptor. Related Articles | Metrics

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THE EFFECTS OF NGF ON THE PROLIFERATION AND DIFFERENTIATION OF ENDOGENOUS NSCs IN THE HIPPOCAMPUS INTO NEURONS AFTER FIMBRIA FORNIX TRANSECTION 2007, 38 (6):  642-646.  doi: Abstract ( )   Objective To study the effects of fimbria fornix transection and intraventricular injection of nerve growth factor(NGF) on the proliferation and differentiation of endogenous neural stem cells (NSCs)in the hippocampus into neurons. Methods Twelve SD rats were divided into the treatment group and control group randomly, six rats in each group. The right fimbria fornix of rats were transected. NGF and artificial cerebrospinal fluid was injected into the lateral ventricle of rats in the treatment group and control group respectively immediately and on the 2nd and 4th day after transection. Bromodeoxyuridin (BrdU) was injected intraperitoneally twice per day from 3 to 7 days after operation. On the 28th day after operation, the brains were perfused for fixation and frozen section. BrdU/NF-200 doublelabel immunofluorescence was used to detect the proliferation and differentiation of NSCs into neurons. Results In dentate gyrus，the number of BrdU positive cells in the transection side was much more than those of normal side both in the treatment group and the control group, and the BrdU positive cells in treatment group were more than those of control group both in the transection side and normal side; The number of BrdU/NF-200 double labeled neurons was the most in the transection side of the treatment group, less in normal side of treatment group and transection side of the control group, but absent in the normal side of the control group.Conclusion Fimbria fornix transection could cause endogenous neural stem cells in the hippocampus proliferate and differentiate into Related Articles | Metrics

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EFFECTS OF NMDA RECEPTORS IN SYNAPSE AND EXTRASYNAPSE OF RAT HIPPOCAMPAL NEURONS ON THE EXPRESSION OF POSTSYNAPTIC DENSITY-95 INDUCED BY AMYLOIDBETA 2007, 38 (6):  647-650.  doi: Abstract ( )   Objective To investigate the mechanism of the soluble Aβ oligomersinduced alteration of synaptic proteins. Methods This study applied immunocytochemistry technique to investigate the changes of the expression of postsynaptic density-95(PSD-95) in primary hippocampal neurons, which was exposed to Aβ25-35 after NMDAR antagonist or agonist treatment. Results The results showed that Aβ25-35 downregulated PSD-95 protein in a dose and timedependent manner. Treatment of cells with MK801 (a general NMDA receptor antagonist) prevented Aβ-induced PSD-95 degradation. Moreover, when extrasynaptic NMDA receptors were blocked by ifenprodil (a NR2B subunit specific antagonist), the Aβ-induced downregulation of PSD-95 was significantly attenuated. Whereas, when synaptic NMDA receptors were blocked by bicuculline (a GABA receptor antagonist) in combination with MK801, the PSD-95 degradation did not change significantly.Conclusion The results suggest Related Articles | Metrics

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A STEREOLOGICAL STUDY OF Age\|RELATED CHANGES OF THE WHITE MATTER AND THE MYELINATED NERVE FIBERS IN THE WHITE MATTER OF RAT BRAIN 2007, 38 (6):  651-655.  doi: Abstract ( )   Objective To explore the changes of the white matter and the myelinated fibers in the white matter of aged rat brain. Methods The white matter and the myelinated fibers in the white matter of 10 young Long\|Evans rats (5 female and 5 male rats, 6\|8-month-old) and 9 aged Long\|Evans rats (5 female and 4 male rats, 18 month old) were quantitatively investigated with stereological techniques and transmission electronic microscopy. Results When compared with those of young rats, the total volume of white matter and the total length of myelinated fibers in the white matter of aged rats were decreased by 24.1% and 41%, respectively. In the white matter of aged rats, the volume density of myelinated fibers, the volume density of myelin sheathes and the mean diameter of myelinated fibers were increased by 30%, 23.9% and 31%, respectively, when compared with those of young rats. However, there were no statistically significant changes of the length density and the total volume of myelinated fibers and the total volume of myelin sheathes in the white matter of aged rats when compared with those of young rats.Conclusion The atrophy of normal aged brain is mainly due to the reduction of the white matter volume. The total length of myelinated fibers in the white matter of aged rats is signigicantly decreased when compared with young rats, which is primarily caused by the loss of myelinated nerve fibers with thinner diameters in the white matter. Related Articles | Metrics

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IDENTIFICATION OF PI3K/Akt PATHWAY RELATED GENES IN VASCULAR SMOOTH MUSCLE CELLS SUBJECTED TO CYCLIC STRAIN 2007, 38 (6):  656-659.  doi: Abstract ( )   Objective To screen out the PI3K/Akt pathway related genes in vascular smooth muscle cells (VSMCs) subjected to cyclic strain. Method VSMCs of rat aorta were subjected to cyclic strain (10 %, 1 Hz) by using a FX-4000T system. Phosphorylation of Akt was examined by Western blotting. Suppression subtractive hybridization (SSH) method was employed to analyze the differently expressed cDNA sequence between the strained VSMCs pretreated with and without wortmannin, a specific inhibitor of PI3K. These fragments were ligated with T vectors, screened through the bluewhite screening system to establish cDNA library, and was sequenced and analyzed in GenBank with BLASTN search.Result Level of Akt phosphorylation of VSMCs was significantly enhanced by the mechanical strain compared with the control. Ten different expressed sequence tags (EST) were gained after sequencing 30 clones randomly selected from 54 white clones. There may be six genes related with the mechanical strain and cell signal PI3K/Akt, such as High mobility group box 1 (HMGB1), Neural precursor cells expressed (Nedd4a), Cyclindependent kinase 5 (CDK5), Histone deacetylase 3 (HDAC3), Ubiquitin-like 1 (Uble1a) and Heterogeneous nuclear ribonucleoprotein H1 (hnRNP). Conclusion SSH is an effective and reliable approach to screen out the genes related with the mechanical strain and PI3K/Akt pathway in VSMCs. Related Articles | Metrics

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THE ROLE OF MAPK IN LPSINDUCED iNOS EXPRESSION IN RAT SCHWANN CELLS 2007, 38 (6):  660-664.  doi: Abstract ( )   Objective To explore the role of mitogenactivated protein kinase(MAPK)in lipopolysaccharide(LPS)induced iNOS expression and NO production in rat Schwann cells by the use of inhibitors PD98059 selective for extracellular signalregulated kinase 1/2(EPK1/2), SB202190 for P38 MAPK and SP600125 for the c-Jun NHSUB>2/SUB>-terminal kinase (JNK). Methods Schwann cells were pretreated with PD98059 (30, 50, 70μmol/L), SB202190 (10, 20, 40μmol/L) and SP600125 (10, 30, 50μmol/L) at the indicated concentrations for 1 hour before the stimulation with LPS (10mg/L) for 4 hour. The estimation of iNOS mRNA, IL-6 mRNA and TNF-α mRNA was performed by RT-PCR; the changes of iNOS protein expression were investigated by Western blotting. The NO level was observed with measurement of nitrite in the cell culture medium. Results LPS could significantly activate MAPK signal pathway and lead to the expression of iNOS and NO production. The iNOS expression and NO production induced by LPS stimulation were significantly inhibited by the three highly specific inhibitors of MAPK. In addition, the inhibitors decreased LPSinduced the expression of IL-6 mRNA and TNF-α mRNA. Conclusion Activation of MAPK pathway is involved in iNOS expression and NO production in rat Schwann cells, and the inhibition of the signal transduction pathway can be effective to reduce Related Articles | Metrics

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THE ATTENUATION OF β-AMYLOID PEPTIDE 25-35-INDUCED Tau HYPERPHOSPORYLATION IN CORTICAL NEURONS BY THE REGULATION OF GINSENOSIDE Rg1 ON THE ACTIVITY OF GSK-3β and PP2A 2007, 38 (6):  665-670.  doi: Abstract ( )   Objective To explore whether ginsenoside Rg1 can attenuate β-amyloid peptide 2535induced Tau hyperphosporylation in rat embryo cortical neurons by regulating the activity of GSK-3β and PP2A. Methods Primary cultures of cortical neurons were prepared from the embryonic day 18±2 in SpragueDawley rats. The experimental groups were designed as follows:1.Neurons culture (control group); 2. Neurons exposed to 20μmol/L Aβ25-35 for 12 hours (Aβ-model group); 3.Neurons exposed to 20μmol/L Aβ25-35 and 10 mmol/L lithium chloride (LiCl), a specific inhibitor of glycogen synthase kinase3β（GSK-3β）, for 12 hours (LiCl group); 4.Neurons exposed to 20μmol/L Aβ25-35 for 12 hours in the presence of 24hour pretreatment with ginsenoside Rg1 (Rg1 pretreatment group) . Western blotting and immunocytochemical staining were used to detect the levels of Tau phosphorylation，total Tau and GSK-3β in cortical neurons. Nonradioimmunoassay was introduced to detect the activity of protein phosphatase 2A (PP2A). Results In Aβ-model group, the levels of Tau protein phosphorylation in the sites of Ser396，Ser199/202，Thr231 and total Ta Related Articles | Metrics

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TRANSPLANTATION OF HUMAN HEPATIC STEM CELLS INCREASED THE EXPRESSION OF ALBUMIN IN HEPATOCIRRHOSIS RATS INDUCED BY CARBON TETRACHLORIDE 2007, 38 (6):  671-674.  doi: Abstract ( )   Objective To study the change of the contents of albumin (ALB) in blood serum and liver tissue after human embryohepatic stem cells transplanted into hepatocirrhosis rats induced by CCl-4. Methods Healthy mature male Wistar rats was randomly divided into control group and hepatocirrhosis group. The control group was injected oily solution with 012ml/100g rat weight, the hepatocirrhosis group was subcutaneously injected 60% carbon tetrachloride oily solution with 0.3ml/100g rat weight.Six weeks after injection, hepatocirrhosis group was subdivided into hepatic stem cells sham group and hepatic stem cells transplanted group. The control group and hepatic stem cells sham group was injected culture media, and hepatic stem cells transplanted group was injected human embryohepatic stem cells 2×106 for each rat. On the first day of 11th weeks, all rats were killed. Blood samples were collected from right ventricles and the contents of ALB in serum of each sample was measured. Observed the expression of human EGFR by immunohistochemistry stain. The expressions of ALB in liver tissue was detected by immunofluorescence and Westernblotting. Results In the control and hepatic stem cells sham group, there was no expression of human epidermal growth factor receptor(EGFR), but in hepatic stem cells transplanted group, there were lots of human EGFR positive cells. In hepatic stem cells transplanted group, but in liver tissue and blood serum the contents of ALB were more than that of hepatic stem cells sham group, but similar to the control group. Conclusion In carbon tetrachloride induced hepatocirrhosis rats, transplanted hepatic stem cells could survive long, and coul Related Articles | Metrics

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THE STUDY OF SPERM PROTEIN sp32/OY-TES-1 mRNA AND PROTEIN EXPRESSION IN HUMAN NORMAL TISSUES 2007, 38 (6):  675-680.  doi: Abstract ( )   Objective To investigate whether the expression of sp32/OY-TES-1 mRNA is various in human normal tissues, study the expression of its protein, and provide more information about sp32/OY-TES-1 expressive profile. Methods Extraction of total RNA from 40 cases of human normal tissues included 19 different types.cDNA was synthesized by reverse transcriptase and tested by conventional polymerase chain reaction (PCR). With the construction of sp32/OY-TES-1 and HPRT plasmid for preparation of standard products, standard curves and the protocol of quantitative real-time PCR (TaqMan method) were generated, and reaction system was optimized. The expressions of sp32/OY-TES-1 and HPRT were tested respectively; OY-TES-1/HPRT was used to evaluate the expression level of sp32/OY-TES-1 mRNA. Sp32/OY-TES-1 protein that expressed in normal tissues was tested by immunohistochemistry staining with polyclonal antibody against OY-TES-1 protein. Results 1.The expressive frequency of sp32/OY-TES-1 in a panel of normal tissues was 72.50% (29/40); 2.The value of OY-TES-1/HPRT was used for the quantification of sp32/OY-TES-1 mRNA expression, which was between 0.000 1 and 0.459 0 in human nontestis normal tissues and showed in the range of logarithmic normal distribution. 3.The relative quantitative expression of sp32/OY-TES-1 mRNA in human testis tissue was 44.9, which was 91-1 000 times higher than that in other normal tissues tested. No positive target protein reaction was found in testis, colon, liver, skeletal muscle, stomach, lens, or leukocyte except in the sperm and the renal tubule of kidney. Conclusion With an extensive expression in human normal tissues, the expression level of sp32/OY-TES-1 mRNA was diverse. Of a panel of normal tissues tested, testis tissue was found wih the highest expression, Related Articles | Metrics

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THE EXPRESSION OF NF-κB p50 AND THE EFFECT OF SAIKOSIDE a IN THE PROCESS OF HUMAN PERIPHERAL BLOOD MONOCYTE DIFFERENTIATING INTO DENDRITIC CELLS 2007, 38 (6):  681-686.  doi: Abstract ( )   Objective To observe the expression of NF-κB p50 protein in the process of the human peripheral blood monocytes differentiating and maturing into dendritic cells. Methods Human peripheral blood monocytes were obtained by discontinuous density gradient centrifuged method. Monocytes of different groups (cytokine group, saikoside group, co\|culture group)were cultured with different stimulating factors such as GM\|CSF, IL-4, TNF-α and saikoside in vitro. The inverted phase contrast microscope, Wright’s staining, immunocytochemical staining were used respectively to observe the changes of morphology, surface differentiation antigen including CD14, CD83, HLA\|DR,S\|100, and the expression of intracellular transcription factor NF-κB p50 protein, in the process of monocytes differentiating into dendritic cells. The image analysis and SPSS software were used to analyze the results. Results The obtained monocytes and dendritic cells possessed their respective cell shape and cell surface antigen marker by cell morphology observation and cell surface marker immunocytochemical staining such as CD14, HLA\|DR, S\|100 and CD83. The monocytes differentiated into immateure dendritic cells(imDC) when cultured with RPMI\|1640 medium containing GM-CSF and IL-4 for 7 days. The imDC differentiatd into mature dendritic cells (mDC) when cultured with RPMI\|1640 medium containing GM\|CSF,IL\|4 and TNFα for another 3 days. The expresion of NF-κB p50 in the two cells showed a significant difference (P<0.05). When Monocytes were cultured with RPMI\|1640 medium containing saikoside a only for 7 days, NF-κB p50 staining in their nucleus was brown\|yellow, but differentiated imDCs was not found. As monocytes were cultured with RPMI\|1640 medium containing GM\|CSF,IL\|4 and saikoside for 7 days, the cells were identified as imDCs by immunocytochemical staining. Then the maturation promoting factor TNF\|α was added, cultured imDC for another 3 days, the nucleus appeared obvious NF-κB p50 brown\|yellow staining which was stronger than that of cytokine group. These cells were identified as mDC. The surface marker expression of co\|culture group stronger than that in cytokine group.Conclusion NF-κB p50 quantity increased in the nucleus when monocytes differentiated into imDC and mDC low concentration of Related Articles | Metrics

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THE EFFECTS OF BRAIN-DERIVED NEUROTROPHIC FACTOR ON THE INDUCE\|DIFFERENTIATED SH\|SY5Y CELLS IN VITRO 2007, 38 (6):  687-691.  doi: Abstract ( )   Objective To study the effects of different contents of brain-derived neurotrophic factor(BDNF) on the expression of cell cycle\|related proteins in G2 phase in differentiated SH\|SY5Y cells in vitro. Methods Sequential exposure of SH\|SY5Y cells to retinoic acid(RA) for 5 days and various contents of brain\|derived neurotrophic factor(BDNF 1μg/L, 10μg/L, 100μg/L) for 2 days yielded homogeneous populations of cells with neuronal morphology. At the same time, control group was incubated in insulin-transferrin selenium-X(ITS) alone for 7 days; RA group was incubated in RA only for 7 days. All media were prepared with DMEM\|F12 containing 1% ITS without serum. Real time one\|step RT\|PCR was carried out to determine the expresson of cell cycle\|related protein in G2 phase. Fluorescence activated cell sorting (FACS) was used to analyze the phase of cell cycle. Results The results were shown that the expression of cyclin B1 was downregulated significantly comparing to RA group and ITS group (P<0.05) in all BDNF groups; The expressions of cyclin B2 and Cdk1 were decreased only in 100μg/L BDNF group comparing to ITS group and RA group(P<0.05)； No change of the expression of Cdk5 was found in all BDNF groups comparing to ITS group. The cell percent of G1 phase in all BDNF groups was increased significantly comparing to RA group and ITS group(P<0.01); the cell percent of S phase was decreased significantly comparing to RA group (P<0.01) in all BDNF groups, and decreased significantly comparing to ITS group in 1μg/L BDNF group and 100μg/L BDNF group(P<0.01).Conclusion The results suggest that BDNF doesn’t enhance the mRNA level of cell cycle\|related proteins of G2 phase, combining the change of cell percent in G1 and S phase certifies that Related Articles | Metrics

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IDENTIFICATION OF LEPTIN GENE FROM HUMAN PLACENTA AND ITS FUSION EXPRESSION IN ESCHERICHIA COLI 2007, 38 (6):  692-696.  doi: Abstract ( )   Objective To identify the sequence of cDNA of human leptin coding region, construct a prokaryotic expression vector, and express human leptin in E.coli. Methods RNA was extracted from human placenta tissue. Leptin gene was amplified from RNA by RT-PCR method. The PCR product was ligated with T vector. The ligation reaction was transformed to E.coli DH5α competent cells. The recombinant plasmid was checked by sequencing and restriction analysis. The human leptin gene was cloned into prokaryotic expression vector PET-28a and tranformed into E.coli BL21. The recombinant strain was constructed and induced by IPTG. The product was analyzed with SDS-PAGE and Western blotting. The expressive product was purified by sodium deoxycholate. Results Analysis indicated that the sequence of human leptin cDNA was the same with the reported sequence. Leptin gene was inserted into prokaryotic expression vector. The fusion protein expressed with high efficiency in recombinant E.coli BL21.The results of SDS-PAGE analysis indicated that the molecular weight of the fusion protein was about 20kD. Conclusion Hunam leptin gene was successfully identified from p Related Articles | Metrics

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THE EFFECTS OF CUMULUS CELL APOPTOSIS ON DEVELOPMENTAL POTENTIAL OF HUMAN OOCYTE IN VITRO 2007, 38 (6):  697-703.  doi: Abstract ( )   Objective To study the effect of cumulus cell apoptosis on the structure of human oocyte cultured in vitro and to probe into the reason why cumulus cell apoptosis rate can be used clinically to predict the developmental potential of oocyte. Methods Cumulusenclosed oocyte complexes (COCs) at the GV stage were cultured in vitro. The apoptosis rate of cumulus cells for every COC was evaluated with HE staining, DAPI staining and TUNEL labeling methods. Oocytes were divided into two groups according to their cumulus cell apoptosis rate. Structure of oocytes was observed under light microscope and transmission electron microscope. Mature oocytes at MⅡ stage were fertilized and cultured in vitro for two days. Then the embryos were evaluated according to their morphology under light microscope.Results The oocytes with low cumulus cell apoptosis rate had a normal structure and a promising developmental potential, while the oocytes with high cumulus cell apoptosis rate had deformed structures and a poor developmental potential. Some deformed oocytes had uneven perivitelline space. For oocytes with a small perivitelline space, the organelle development was slow compared with those with a large perivitelline space. Some had unevenly piled organelles. Secondary lysosomes and large space probably caused by the decopmounding of lysosomes were found in the ooplasm of these oocytes. The deformed oocytes appeared to have lots of swollen mitochondria with blurred cristae and membrane. Some of these oocytes had fractured first polar bodies and abnormally thick or thin zona pellucida. The cell junctions and microvilli of cumulus cells reduced as well. Conclusion The effect of cumulus cell apoptosis on the structure of oocyte cultured in vitro was revealed. The reaso Related Articles | Metrics

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REVERSAL OF RESISTANCE TO ADRIAMYCIN IN MCF-7/ADM CELLS BY REALGAR IN COMBINATION WITH β-ELEMENE 2007, 38 (6):  704-707.  doi: Abstract ( )   Objective To study the reversal effect of adriamycin (ADM) resistance in human breast cancer cell line MCF-7/ADM by realgar (REA) combined with β-elemene (β-ELE), two types of anticancer drugs. Methods The sensitivity to ADM of MCF-7/ADM cells was studied by MTT assay.The intracellular accumulation of ADM in MCF-7/ADM cells was observed by fluorescentspectrophotometry. Expressions of P-GP protein and Bcl-2 protein were detected by flow cytometry. Results The combined treatment of REA and β-ELE could significantly enhance the cytotoxic effect of ADM on MCF-7/ADM cells to 4.2 fold as compared with the treatment of REA or β-ELE respectively (P<0.01). The intracellular accumulation of ADM in MCF-7/ADM cells was significantly increased after the combined treatment (P<0.01). The expression of P-GP protein in MCF-7/ADM cells was reduced from(95.90±3.02)% to (76.50±5.16)%(P<0.01) while the expression of Bcl-2 protein was reduced from(90.20±3.99)% to (50.00±1.63)%(P<0.01).Conclusion The combined treatment of REA and β-ELE could reverse more strongly the drug resistance to A Related Articles | Metrics

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EFFECT OF AcSDKP ON THE COLLAGEN SYNTHESIS AND EXPRESSION IN CULTURED RAT CARDIAC FIBROBLASTS MEDIATED BY TRANSFORMING GROWTH FACTOR BETA1 2007, 38 (6):  708-712.  doi: Abstract ( )   Objective To investigate whether AcSDKP can inhibit collagen synthesis in cultured rat cardiac fibroblasts mediated by transforming growth factor beta1(TGF-βSUB>1/SUB>).Methods Neonatal rat cardiac fibroblasts were isolated. The synthesis of collagen was measured by SUP>3/SUP> Hproline incorporation assay. The expressions of type Ⅰ and Ⅲ collagen protein were detected by immunocytochemistry and Western blotting. Results SUP>3/SUP> Hproline content increased gradually (147.6±10.2， 229.2±16.4，427.0±40.6，454.8±26.1 CPM) and was 1.61， 2.50，4.66 and 4.97 times of control group(91.6±9.8 CPM) respectively in rat cardiac fibroblasts mediated by TGF-βSUB>1/SUB> concentration of 1μg/L, 2.5μg/L, 5μg/L and 7.5μg/L. Compared with TGF-βSUB>1/SUB> group (5μg/L), SUP>3/SUP> Hproline content decreased gradually(378.8±6.4，292.8±14.4，130.6±17.6 and 230.6±19.4 CPM) and was 88.7%, 68.6%， 30.6% and 54.0% of TGF-β1 group with increasing of AcSDKP concentration(10SUP>-10/SUP>mol/L, 5×10SUP>-10/SUP>mol/L, 10SUP>-9/SUP>mol/L, 10SUP>-8/SUP>mol/L). Compared with control group, the expression of collagen type Ⅰ and type Ⅲ in rat cardiac fibroblasts increased significantly in TGF-βSUB>1/SUB> group(5μg/L). Obsorbance values Related Articles | Metrics

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SAFETY TEST OF GOSSYPOL PLUS METHYLTESTOSTERONE AND ETHINYLESTRADIOL AS A MALE CONTRACEPTIVE 2007, 38 (6):  711-717.  doi: Abstract ( )   Objective To investigate whether administration of gossypol of 12mg/kg combined with methyltestosterone 20mg/kg and ethinylestradiol 100μg/kg daily for a long term of 24 weeks could affect rat spermatogonial stem cell function. Methods TdTmediated dUTP nick endlabeling (TUNEL) detection, spermatogonial stem cell transplantation and fertility recovery evaluation were carried out. Results The administration of this regimen could induce germ cell going apoptosis; the apoptotic signal positive cells were spermatocyte and spermatid, no spermatogonium was found to be signal positive. Two to three months after transplantation of medicine treated spermatogonial stem cells, elongated rat spermatids were identified in the seminiferous tubules of recipient nude mice. Six weeks after withdrawal of the administration, fertility of the medicine treated rats was recovered completely compared with that of the control group. After a series of tests, litters of the first generation showed to be normal. Conclusion These results indicated that the administration of this regimen would not affect the function of rat spermatogonial Related Articles | Metrics

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MOLECULAR MECHANISMS OF ESTROGEN INDUCED APOPTOSIS OF SYNTHETIC VASCULAR SMOOTH MUSCLE CELLS： p38 PATHWAYDEPENDENT UPREGULATION OF MYOCYTE ENHANCER FACTOR 2 2007, 38 (6):  718-721.  doi: Abstract ( )   Objective To explore the underlying molecular mechanisms of estrogen induced apoptosis of synthetic vascular smooth muscle cells (VSMCs). Methods Cultured VSMCs on passage 3 were divided into three groups: 17β-estradiol (E\-2) group, p38 inhibitor group (SB+E\-2) and control group. With treatment for 72 hours,the apoptosis of cells was quantified by measuring the intracellular nucleosomes with ELISA; expressions of total p38, phospho-p38 and myocyte enhancer factor2 (MEF2) were analyzed by Western blotting, with a confirmative immunohistochemical staining for MEF2. Results Compared with that of control cells, the rate of apoptosis in VSMCs grown in E\-2 was significantly increased, with upregulations of total p38, phosphop38 and MEF2. These effects of E\-2 were blocked in SB+E\-2 group in which the rate of cell apoptosis and expression of p38 had no obvious change compared with that of control group, while the expression of p-p38 and MEF2 was decreased. Immunohistochemical staining showed that MEF2 was mainly located in cellular nucleus, and its expression was increased in E\-2 group but decreased in SB+E\-2 group. Conclusion Estrogen may induce apoptosis of synthetic VSMCs Related Articles | Metrics

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EXPRESSION AND LOCALIZATION OF AQUAPORIN 1 IN MOUSE SEMINAL VESICLE AND PROSTATE GLAND DURING POSTNATAL DEVELOPMENT 2007, 38 (6):  722-726.  doi: Abstract ( )   Objective To examine the expression and distribution of Aquaporin 1 （AQP1） in the seminal vesicle and prostate gland of male mice during postnatal development. Methods The RT-PCR for AQP1 mRNA, Western blotting for protein, and immunohistochemistry for distribution in the seminal vesicle and prostate gland on postnatal day 15, day 35 and day 70 were carried out. Results The results of RT-PCR and Western blotting showed that the level of AQP1 mRNA and protein were significantly increased in the seminal vesicle and prostate gland on postnatal day 70 (P<0.05). The positive signals of AQP1 protein were localized immunohistochemically in the smooth muscle cells of seminal vesicle and the basement membrane of glandular epithelium and the mesenchyme cells of prostate gland on postnatal day 15 and day 35, but it was hardly detected in the epithelium of seminal vesicle. Thereafter, AQP1 protein was intensively expressed in the epithelium of seminal vesicle and prostate gland on postnatal day 70.Conclusion The different expressions and distributions of AQP1 mRNA and protein in the seminal vesicle and prostate gland of mice in different ages may be related to in the development of male accessory glands. Related Articles | Metrics

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EFFECTS ON SPERM OOCYTE PENETRATING BY ANTI SERUM TO HUMAN SPERM MANNOSE RECEPTOR 2007, 38 (6):  727-730.  doi: Abstract ( )   Objective To study the effect of mannose receptor(MR) from human spermatozoa on spermegg fusion. Methods The Balb/c mouse was immunized with the purified mannose receptor(pMR) which was isolated from motile human sperm by mannoseagarose gel affinity chromatography and the antiMR serum was harvested. Human sperm after induction of acrosome reaction was pretreated with antiMR serum and subsequently processed in sperm penetration assay (SPA). Results The antiMR serum can inhibit human spermatozoa from binding and penetrating zonafree golden hamster eggs with a dosedependent reduction of Fertilization rate(FR) and Penetration index(PI).Conclusion Related Articles | Metrics

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IMMUNOHISTOCHEMICAL STUDIES OF EGF AND IL-2 POSITIVE CELLS IN RAT SUBMANDIBULAR GLAND DURING HEROIN DEPENDENCE PERIOD 2007, 38 (6):  731-735.  doi: Abstract ( )   Objective To observe the effect of heroin on EGF and IL-2 positive cells in the submandibular gland of rats. Methods Normal SD rats were divided into normal control group (NGC),saline control group (SCG)and heroin dependence group (HDG). The heroin dependent model of rats was established by subcutaneous injection of heroin, and submandibular glands were excised on the following days respectively: the 10th, 17th, 24th, 31th and 38th. Immunohistochemical SABC method, image analysis and cells calculation were used in the study. BR> Results 1. As compared with those of the NCG, the number of EGF and IL-2 positive cells and the intensity of staining in SCG didn’t change obviously. 2. During heroin dependence, the immunohistochemical staining showed that the number of EGF and IL-2 positive cells increased obviously(P<0.05); meanwhile, the results of image analysis showed that the mean grey degree of EGF and IL-2 positive cells were lower than that of NCG and SCG groups. 3.The colocalization of EGF and IL-2 was proved in the submandibular gland of rats.Conclusion The number of EGF and IL-2 positive cells and the intensity of staining change regularly in the submandibular gland of rats. It suggestes that the EGF and IL-2 from submandibular gland of rat were involved in the regulating proc Related Articles | Metrics

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EXPRESSION AND EFFECT OF L-SELECTIN IN MICE OVIGERMS DURING THE PERIIMP LANTATION PERIOD 2007, 38 (6):  736-740.  doi: Abstract ( )   Objective To investigate the expression of L-selectin mRNA and protein in pregnant mice during the periimplantation period and the effect of L-selectin in the embryo implantation. Methods 1.Mice ovigerms were divided into 4 groups: 1-cell stage, 4-cell stage, morula stage and blastocyst stage. The expression of L-selectin mRNA and protein in mice ovigerms was detected by reaL-time fluorescence quantitative PCR(FQ-PCR) and immunofluorescence histochemistry. The location of L-selectin was analyzed by immunofluorescence histochemistry; 2. Mice were divided into 4 groups: experiment group 1(1∶50), experiment group 2(1∶100), experiment group 3(1∶200) and control group (without any intervention to mice cornu uteris). On the morning of day 4 of pregnancy female mice were injected with 5μl of the L-selectin monoclonal antibody into one horn(experiment side) of uteri and with 5μl normal saline into another horn(control side) in the game mouse. The number of embryos implanted was counted and recorded on day 8 of pregnancy to study the effects of the L-selectin on embryo implantation in vivo. The data were analyzed by analysis of variance. Results 1. With the development and maturation of the ovigerm, the expression of L-selectin mRNA and protein was increased gradually(P<0.05), and the level of L-selectin mRNA on blastocyst stage was higher than those of other groups(P<0.05). There was a little expression of L-selectin protein in 1-cell stage and 4-cell stage. The expression of L-selectin protein which was mainly distributed around the morula increased significantly. The level of L-selectin protein which was mainly distributed on the cell membrane of trophoblast was the highest during the periimplantation period(P<0.05); 2. The number of embryos implanted in the horns treated with L-selectin monoclonal antibody was smaller than those in corresponding controls(P<0.05).Conclusion L-selectin of the ovigerm might particip Related Articles | Metrics

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THE STUDY OF INFLUENCE OF LONGTERM SMOKING ON THE EXPRESSION AND MUTATION OF p53 AND K-ras GENE IN RAT LUNGS 2007, 38 (6):  741-745.  doi: Abstract ( )   Objective To observe the influence of longterm smoking on the expression of p53 and K-ras in rat lung tissues, and to study the relationship of smoking to the mutation of p53 and K-ras gene. Methods The model of Wistar rat smoking was built up. Seventytwo healthy male Wistar rats were divided into the experimental group and control group at random. The rats of the experimental group were compelled to smoke, and the rats of the control group were given the same condition as the experimental group was, without smoking. The rats of the experimental group were smoked for 6 months. At the end of each month, 6 rats were chosen from the two groups respectively, their lung tissues were sampled and immunohistochemistry was applied to observe the expression of p53 and K-ras in lung tissues. Finally, the mutation which might happen in the exon 5, 6, 78 of p53 and the exon 1 of K-ras was examined by PCRSSCP. Results The p53 protein was expressed in cell nucleus and K-ras in cytoplasm. The positive ratio of protein expression was increased with the extension of smoking time. The mutation of p53 was increased as the smoking time extended. But the effect of smoking time was not that significant on the mutation of K-ras.Conclusion Smoking can strengthen the expression of p53 and K-ras protein and can also result in gene mutation. As the time of smoking extended, those phenomenons were tending to rise. That provided the theoretical evidence which can be used to judge the lesion of lung tissues caused by smoking and help the early diagnosis of smokingrelated lung carcinomas. It Related Articles | Metrics

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SITES OF ACTION OF COMBINATION REGIMEN OF LOW-DOSE GOSSYPOL ACETIC ACID WITH STEROID HORMONES ON MALE RAT 2007, 38 (6):  746-750.  doi: Abstract ( )   Objective To investigate the target sites for male rats infertility by administered with combination regimen of lowdose gossypol acetic acid (GA) with steroid hormones (desogestrel/ethinylestradiol/testosterone undecanote (DSG/E/TU). Methods Adult male rats were randomly divided into four groups. Group GH: rats were fed orally with GA (125mg(kgI>&#/I>8226;day)) and DSG (0.125mg/(kgI>&#/I>8226;day))/E(0.025mg/(kgI>&#/I>8226;day))/TU (100mg/(kgI>&#/I>8226;day)); Group G: rats were administered with a single dose of GA (12.5mg/(kgI>&#/I>8226;day)); Group H: DSG(0.125mg/(kgI>&#/I>8226;day))/E(0.025mg/(kgI>&#/I>8226;day))/TU (100mg/(kgI>&#/I>8226;day)) were administered to rats; Group C: rats only received vehicle (1% methyl cellulose). The testis and epididymis were removed at 4, 6, 8, 10 weeks respectively. Morphology of testis, epididymis and sperm were observed, and testicular sperm number was counted, epididymal sperm motility was measured. Results We found that group GH had similar results as group H ,but not either as group G or as group C, in testicular sperm count and the morphology of testis seminiferous tubules, while in epididymis, the sperm motility decreased and the abnormal sperm increased obviously in all three treatment groups compared with group C.Conclusion The steroid hormones are the main functional factors on spermatogenesis in testis, gossypol and the hormones have synergetic effects on sperm motility in epididymis. With either independent or synergistic actions of gossypol and steroid hormones, the combined regimen have a promising effect on male rat contraception. Related Articles | Metrics

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EXPRESSIONS OF ANGIOPOIETHIN 1 AND 2 IN THE ENDOMETRIUM OF WOMEN WITH ABNORMAL BLEEDING AFTER MEDICAL ABORTION 2007, 38 (6):  751-754.  doi: Abstract ( )   Objective To investigate the expression and the roles of Angiopoietin 1(Ang-1) and Angiopoietin 2(Ang-2) in the endometrium of women with abnormal bleeding after medical abortion. Methods Analyzing endometrial pathological change of 1087 patients with abnormal bleeding after medical abortion in early pregnancy,and the endometrial specimens from 40 patients were randomly chosen for the study. The endometrial specimens from 20 women without abnormal bleeding after medical abortion were used as control group.Immunohistochemistry was used to detect the levels of Ang-1 and Ang-2 proteins in endometria. Results The percentage of patients with both the residul decidua and villus was 80.5%; positive immunoreactive signals of Ang-1 and Ang-2 were found in the endometrial glandular epithelium, stromal cells and the endothelial cells of vessels; the expression rate of Ang-1 and Ang-2 in the patients with abnormal bleeding were higher than that in the control group（P<0.01）.Conclusion The residul decidua and villus are probably the main cause of abnormal bleeding after medical abortion.The hyperexpression of Ang-1 and Ang-2 in the endometrium of patients with abnormal bleeding after medical abortion may play an important role in the process Related Articles | Metrics

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EXPRESSION OF Bcl-2 AND Bax PROTEINS IN THE DEVELOPING SMALL INTESTINE OF HUMAN FETUS 2007, 38 (6):  755-758.  doi: Abstract ( )   Objective To explore the role of cell proliferation and apoptosis, and the expression significance of involved protein as Bcl-2 (B-cell lymphoma/leukemia-2) and Bax (BcL-associate X protein) in the developing small intestine of human fetus. Methods The expression product of Bcl-2 and Bax was investigated with immunohistochemical methods in the 2nd, 3rd and 4th months of gestation respectively. Results In the second, the third and the fourth month of gestation, the Bcl-2 immunoreactive positive signals were found in the ganglion cells of intermuscular and submucous nerve plexus. Bax positive cells were observed in the cytoplasm of the simple columnar epithelium cells of the small intestinal mucous layer.Conclusion Bcl-2 and Bax proteins regulate the developing small intestine of human fetus. Related Articles | Metrics

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MICROANATOMY OF THE Labb VEINS AND THEIR VENOGRAMS OBTAINED BY IMAGEOLOGY 2007, 38 (6):  759-764.  doi: Abstract ( )   Objective To correlate microanatomy of the Labbé vein with its venograms in order to provide a basis for the preservation of the Labbé veins in the supratentorial surgical approaches. Methods A total of 30 human cadavers (60 sides) and 61 patients (110 sides) were examined in this study. Each head of the cadavers were injected with bluecoloured latex via the superior sagittal sinus and internal jugular veins. The venograms of each patient were obtained from the venous phases of Digital Subtraction Angiography (DSA) (60 sides out of 36 patients) or Computed Tomographic Venography (CTV) (50 sides out of 25 patients). Results Based on the location of the dural entrance where a vein enters a dural sinus, the Labbé veins were divided into four groups: transverse sinus group, tentorial group, petrosal group and uppertransverse sinus group. In the transverse sinus and petrosal groups, the Labbé veins drained into transverse sinus directly,while indirectly in the tentorial and uppertransverse sinus groups, via the tentorium sinus or the uppertransverse sinus, which was a kind of meningeal veins running in the two layers of cerebral dura matter. The length of the meningeal veins was(10.0±7.2)mm (1.7-23.6mm). The Labbé veins were mainly located around the STP junction which is at the confluence of sigmoid sinus, transverse sinus and superior petrosal sinus. The distance between the dural entrance of the Labbé vein and STP junction was (16.8±10.2)mm (0-40.1mm). Compared with the observation in cadavers, there was no difference in DSA and CTV examinations.Conclusion The preoperative venogram is useful to design the individualized surgical approach for the preservation of the Labbé veins. In general, the medium supratentorial approach has the l Related Articles | Metrics

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PRIMARY CULTURE AND IDENTIFICATION OF MOUSE RENAL TUBULAR EPITHELIAL CELLS 2007, 38 (6):  765-767.  doi: Abstract ( )   Objective To establish a method for the primary culture, subculture and identification of mouse renal tubular epithelial cells(mRTECs). Methods Renal tubular segments sticking and three kinds of digesting(trypsin,trypsin+EDTA, collagenase Ⅰ) were used in the primary culture of mRTECs respectively.Cell vitality, morphology and quantity were observed and evaluated by phase-contrast microscopy after trypan-blue staining. Trypsin digestion was used in the subculture of mRTECs. The types of cells were identified by cellular immunocytochemistry. Results The culturing methods of renal tubular segments sticking and collagenase Ⅰ digesting were successfully used in the primary culture of mRTECs and the latter was quicker for the growth of cells.Conclusion Collagenase Ⅰ digesting is an ideal method of cellular culture for the primary culture of mouse renal tubular epith Related Articles | Metrics

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STUDY ON THE PROTECTIVE SOLUTIONS OF hRPE CELLS AFTER BREAKING AWAY CULTURE CONDITION IN VITRO 2007, 38 (6):  768-770.  doi: Abstract ( )   Objective To investigate the best protective solution that could maintain the cell activity and prolong the survival time of the human retinal pigment epithelium (hRPE) cells after break away in vitro culture case. Methods hRPE cells (stored in our laboratory) culture in DMEM/F12, added 10% FBS, EGF 20μg/L and bFGF 20μg/L, which had been cultured for more than five generations. When these culture hRPE cells had been confluenced more than 80%, the hRPE cells were harvested and placed into six kinds of protective solutions (balanced salt solution, Quinn’s advantage medium, physiological salt solution, 18amino acid solution, 5% glucose solution, artificial cerebrospinal fluid), then the cells were stained with the Annexin V /FITC, and the numbers of cell death and cell apoptosis were detected with the fluorescence activated cell sorter (FACS). The hRPE cells were in six kinds of protective solutions and were investigated at three different times from 2 hours to 8 hours. Results The result presented as follows: At the room temperature, the hRPE cells in all six kinds of protective solutions occurred necrosis within 30 minutes. To investigate the state at 4℃, in a duration of 2 hours, the apoptosis rate in the group of balanced salt solution was lowest (0.9%); As compared with the group of balanced salt solution, the apoptosis rate of artificial cerebrospinal presented a great difference (P0.05). The apoptosis rate in the group of artificial cerebrospinal fluid increased from 26.74% to 41.36% in a duration of 4 to 8 hours. The death rates in all six solutions were approximately the same with the apoptosis rate. Conclusion The preservative liquid of balanced salt solution at 4℃, as compared with the other 5 kinds of protective solutions, was the most suitable condition for the hRPE cells to maintain Related Articles | Metrics