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2010, Volume 41 Issue 4
06 August 2010 Previous Issue    Next Issue
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Antioxidation effects and mechanism of Gastrodin in AD rat models 2010, 41 (4):  485-490.  doi: 10.3969/j.issn.0529-1356.2010.04.001 Abstract ( )   To established β-amyloid peptide (1-40) (Aβ SUB>1-40/SUB>) and D-galactose(D-gal) AD rat models and to explore the anti-oxidation molecular mechanism of the Gastrodin and provide the experimental foundations for clinical treatment of AD. AD rat models were established by lateral ventricle injection of Aβ1-40 and abdominal cavity injection of D-gal (100 mg/kg) to thirty six male SD rats. Meantime, the rats were treated by intragastric administration the Gastrodin. Then the behavioral testing of experimental rats was performed by the Morris water maze(MWM). The thiol antioxidants including hydrogen peroxide (HSUB>2/SUB>O SUB>2/SUB>), glutathione reductase (GR) and glutathion (GSH) activities were examined by colorimetric method. The concentration of the p38 and P-p38 was examined respectively by Western blotting. The AD model rats when compared with control group exhibited a significant increase in escape latencies (EM> P/EM> <0.05), and a decrease in the time of staying at the third quadrants of platform and the degree of crossing over a platform. The cerebral cortex concentration of the H2O2 was increased, and the concentrations of the GR and GSH were decreased ( EM>P /EM><0.05). The expression of P-p38 was increased (EM> P/EM> <0.05). After the treatment with Gastrodin, the AD model rats exhibited a significant decrease in escape latencies ( EM>P /EM><0.05), an obvious increase in the time of staying the third quadrants of platform ( EM>P/EM> <0.05) and the increase of crossing over a platform (EM> P/EM> <0.05) when compared with the AD group ( EM>P /EM><0.05). The concentration of the H2O2 was decreased, the concentrations of GR and GSH were increased ( SUP>P /SUP><0.05). The expression of the P-p38 was less than that of the AD model ( EM>P /EM><0.05). But there were no significant differences between the three groups in the expression of the p38 ( EM>P/EM> >0.05). Gastrodin could improve the oriented learning and memory capacity and prevent the neurodegeneration of central nervous systems in AD model rats by partly affecting the expression of the thiol antioxidants(e.g. GR and GSH) and the expression o Related Articles | Metrics

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Pathway of phospholipase C-inositol trisphosphate is involved in the increase of ［CaSUP>2+/SUP>］i induced by β-amyloid peptide (25-35) in hippocampal neurons 2010, 41 (4):  491-497.  doi: 10.3969/j.issn.0529-1356.2010.04.002 Abstract ( )   Objective To investigate the effect of endoplasmic reticulum(ER) calcium stores on calcium overload induced by β-amyloid peptide (25-35)(AβSUB>25-35/SUB>) in hippocampal neurons. Methods The hippocampal neurons were loaded with calcium-sensitive fluorescent indicator Fluo-3/AM.Intracellular calcium concentration (［CaSUP>2+/SUP>］i)changes were measured in different conditions of drug intervention by using laser scanning confocal microscopy. Results Groups of 2, 10, 20μmol/L AβSUB>25-35/SUB> all increased ［CaSUP>2+/SUP>］i in hippocampal neurons(EM>n/EM>=5, EM>P/EM><0.05). 2-APB, an inhibitor of ER CaSUP>2+/SUP> release through channels associated to inositol trisphosphate receptor(IP3R), was shown to prevent the aggregated AβSUB>25-35/SUB>induced rise of ［CaSUP>2+/SUP>］i, suggesting the involvement of CaSUP>2+/SUP>released by ER. However, dantrolene, an inhibitor of ER CaSUP>2+/SUP> release through channels associated to ryanodine receptor(RyR), could not prevent the rise of ［CaSUP>2+/SUP>］i induced by AβSUB>25-35/SUB>. Treatment with aggregated AβSUB>25-35/SUB> for 1 hour induced a significant decrease in the ER CaSUP>2+/SUP> content (EM>P/EM><0.01), which was more pronounced 24 hours after the addition of aggregated AβSUB> Related Articles | Metrics

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Radial glial cell and development of cerebellar cortex in mouse 2010, 41 (4):  498-503.  doi: 10.3969/j.issn.0529-1356.2010.04.003 Abstract ( )   Objective To investigate the differentiation of radial glial cells during the development of mouse cerebellar cortex. Methods The immunofluorescent stainings and 5-bromodeoxyuridine(BrdU) assay were used to label neural stem cells, radial glial cells, Purkinje cells and granule cells in cerebellum(57 cases，which were divided into 19 groups,and each had 3 mice) from embryonic day 8 (E8) to postnatal day 180 (P180). Results The radial glial cells could be found in neuroepithelium at E13 Then they differentiated various neurons and Bergmann cells. The radial glia cells played very important role in the cerebellar lamination. Conclusion Radial glial cells are generated from the neuroepithelial cells, and they act as the neural progenitors for neurons and neuroglia. During the development of cerebellar cortex, radial glial cells can differentiate into Purkinje cells and granule cells and provide the pathways and scaffolds for the neuron migration. Related Articles | Metrics

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Change of Toll-like receptor 4 expression in microglial cells after spinal cord compression injury in rats 2010, 41 (4):  504-509.  doi: 10.3969/j.issn.0529-1356.2010.04.004 Abstract ( )   Objective To explore the change of Toll-like receptor 4(TLR4) expression in microglia after compression injury in the spinal cord of rats, and to compare the distribution of TLR4 positive cells with the area of injury and immunoglobulin G (IgG) extravasation at different time points. Methods Spinal cord compression injury was made at T8 segment in 48 adult male SD rats, and the rats were randomly divided into 6 groups. Sham and injured animals were fixed by perfusion of 4% paraformaldehyde, 2 cm of the spinal cord blocks with compressed site in center were then removed at 0 hour, 3 hours, 24 hours, 72 hours and 7 days after injury. Frozen sections were made for HE and immunofluoresent staining to detect the injury area and the expression of TLR4 in the spinal cord. The distribution of TLR4 positive cells, the injury area and the extravasated IgG were compared as well. The number of TLR4 positive microglial cells was counted by using Image Pro Plus 6.0 software. BR> Results TLR4 was mainly expressed in microglial cells and the expression started to increase between 3 hours and 24 hours, and peaked at 72 hours after the spinal cord compression injury, then descended at 7days, according to the number of TLR4-positive microglia. The distribution of TLR4 immunoreactive product was consistent with the area of injury and that of extravasated IgG. Conclusion In the rat model of spinal cord compression injury, TLR4 expressed in microglial cells may play an important role in the secondary injury and TLR4 expression may associate with the compromise of blood-spinal cord barrier. BR> Related Articles | Metrics

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Changes of phosphorylated extracellular signal-regulated kinase 1/2 in the medial prefronotal cortex of chronic forced swimming model rats 2010, 41 (4):  510-513.  doi: 10.3969/j.issn.0529-1356.2010.04.005 Abstract ( )   Objective To observe the changes of phosphorylated extracellular signal-regulated kinase 1/2 (pERK1/2) in the medial prefrontal cortex (mPFC) of chronic forced swimming rats. Methods Thirty male Wistar rats were randomly divided into two groups with fifteen in each. The chronic forced swimming stress (CFSS) model animals were given swimming stress for 15min once daily for 28 days, and the controls were free of stress. The depression behavior was examined using sucrose preference test, openfiled test and Morris water maze. The expression of pERK1/2 was detected using immunohistochemistry and Western blotting. Results The consumption of sucrose and preference of sucrose in CFSS group were (4.114±0.644)%, (86.610±4.450)% respectively, those in control group were (8.157±1.105)%, (94.930±2.893)% (EM>P/EM><0.01). The erect quantity in CFSS group and control group were 1.75±0.96, 6.00±0.82 respectively (P<0.05), and the escape latency in two group were (20.762±3.236)s, (5.632±1.065)s (EM>P/EM><0.01). The result of immunohistochemistry analysis showed that the integral absorbance of pERK1/2positive cells in control group and CFSS group were 47.594±5.355 and 110.810±10.643 respectively (EM>P/EM><0.01). Western blotting showed that relative expression Related Articles | Metrics

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Expression of nuclear receptor related factor 1 in the developing rat brain 2010, 41 (4):  514-518.  doi: 10.3969/j.issn.0529-1356.2010.04.006 Abstract ( )   Objective To observe the dynamic changes in protein expression of the orphan nuclear receptor related factor 1(Nurr1) as well as its correlation with proliferating cell nuclear antigen(PCNA) in the developing rat brain, and to investigate the significance of Nurr1 in the differentiation and migration of neural cells. Methods Paraffinembedded sections, immunohistochemistry, double labeling immunohistochemical methods were adopted. Results Nurr1 was expressed in small quantities in the cells surrounding the lateral ventricle. Along with the maturing of the cells, the positive expression increased apparently and differentiated and migrated to the distant sites from the lateral ventricle. In postnatal rats, the positive cells surrounding the lateral ventricle decreased remarkably. High levels of the expression was found in the cerebral cortex on the postnatal day 1-5, then along with the maturing of the cells, the positive expression decreased obviously. Positive cells were rare in matured cerebral cortex. The comparative observation against PCNA showed that the two were expressed at different sites and in different cells. Nurr1 was mainly expressed in differentiated and migrated immature cells, but not in proliferating cells. Conclusion It implies that Nurr1 may play a regulatory role in the differentiation, migration and maturation of the rat neural cells. Related Articles | Metrics

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Effects of estrogen on cognitive functions and on the neurons in CA1 region of hippocampus of the SAMP8 mouse 2010, 41 (4):  519-523.  doi: 10.3969/j.issn.0529-1356.2010.04.007 Abstract ( )   Objective To investigate the effects of estrogen on the cognitive function and on the hippocampus neurons of SAMP8 mouse. Methods Forty five 6-month-old female SAMP8 were divided into three different groups: sham group, OVX group, OVX+E group, and SAMR1 mice were used as comparision of homologization. Morris water maze test was used to test the ability of learning and memory. HE staining and immunohistochemistry technique were used to reveal the neurons and neuronal nitric oxide synthase(nNOS) positive neurons in the CA1 subregion. Flow cytometry was used to detect the quantities of the nNOS in the hippocampus. Results The escape latency of the OVX group was that of the sham group（EM>P/EM><0.05）and the total number of crossing the platform of the OVX group was fewer than that of the sham group（EM>P/EM><0.05）. Estrogen replacement therapy could improve the learning and memory ability（EM>P/EM>>0.05）; The OVX group of the CA1 was significantly changed, and the nNOS positive neurons, average absorbance and the nNOS fluorescence index (FI) of the OVX group of the CA1 were shorter than that of the sham group（EM>P/EM><0.05）. The OVX+E group and the sham group showed no difference in results（EM>P/EM>>0.05）. Conclusion The present study suggests that estrogen can improve the ability of learning and memory, protect Related Articles | Metrics

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Influence of nicotine on dopamine transporter and tyrosine hydroxylase in rat brain 2010, 41 (4):  524-527.  doi: 10.3969/j.issn.0529-1356.2010.04.008 Abstract ( )   Objective To study the effect of nicotine on the dopaminergic system by analyzing the altered expression of dopamine transporter(DAT) and tyrosine hydroxylase(TH) in the brain of nicotine-treated rats. Methods Healthy male Wistar rats were selected and then injected for 7 days with 0.4 mg/kg nicotine every day. Immunohistochemistry and Western blotting were used to detect the altered expression of DAT and TH in the dopaminergic system in different brain regions of the nicotine treated rats. Results Compared with the control: 1. Immunohistochemistry showed the mean gray value of DAT in nucleus accumbens (NACC) and ventral tegmental area(VTA) decreased 12.43 % and 12.85 % respectively, the mean gray value of TH in NACC and VTA decreased 11.87 % and 10.09 %. 2. Western blotting showed the ratio of relative absorbance of DAT/β-actinin in CPu-NACC and SN.VTA of the the nicotine-treated rats increased 75.68 % and 117.14 % respectively; the ratio of relative absorbance of TH/β-actinin in caudate putamen(CPu)-NACC and substantia nigra(SN)-VTA increased 66.32 % and 60.31 % respectively. Conclusion Nicotine treatment increases the expression of DAT and TH in rat brain, which may have some relations with the mechanism of nicotine dependence. Related Articles | Metrics

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Expression of Caspase-3 in rat hippocampus during postnatal development 2010, 41 (4):  528-531.  doi: 10.3969/j.issn.0529-1356.2010.04.009 Abstract ( )   Objective To investigate the relationship between active Caspase-3 and apoptosis in Wistar rat hippocampus during postnatal development. Methods Immunofluorescent staining was applied to observe the expression of active Caspase-3 and Hoechst 33342 in the CA1, CA3 and dentate gyrus（DG）of rat hippocampus during postnatal development. Results The expression of active Caspase-3 reached a peak at postnatal day 7(P7) in CA1, at P2 in CA3, then decreased with age. Whereas in DG, active Caspase-3 expression increased slightly after P7, reaching a maximum at P14, and remained at high levels for the rest of the investigated period. In addition, the number of apoptotic cells in the three regions all reached maximum levels at P7, then decreased with age. Conclusion There are specific spatiotemporal patterns of expression of active Caspase-3 in the postnatally developing rat hippocampal subregions and active Caspase-3 in the postmitotic neurons of CA1 and CA3 and in neuronal progenitor cells of DG may have distinct roles and mechanisms Related Articles | Metrics

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Role of Bmi-1 in the self-renewal and proliferation of neural stem cells isolated from human corpus striatum 2010, 41 (4):  532-537.  doi: 10.3969/j.issn.0529-1356.2010.04.010 Abstract ( )   Objective To discuss whether Bmi-1 is participated in maintenance of the self-renewal and proliferation of neural stem cells (NSCs) isolated from fetal striatum. Methods Neural stem cells were transfected with Bmi-1 targeted shRNA-lentiviral vectors and GFP control vector respectively. At 0 hour,48 hours, 72 hours, and 168 hours after transfection, the Bmi-1 mRNA expression level was measured by Realtime PCR. The numbers and diameters of newly formed neurosphere were calculated under microscope. Results The transfection efficiency was shown to be more than 90% by expression of reporter gene GFP. At 0 hour,48 hours, 72 hours and 168 hours after transfection, transcription level of Bmi-1 were to be （107.3±8.0618）%, （32.7±9.74）%, （24.4±12.15）%, （32.7±10.39）% compared with 0 hour, the relative expression levels of Bmi-1 at 48 hours, 72 hours, and 168 hours were significantly decreased(EM>P/EM><0.05). Accordingly, The number and size of newly formed neurospheres were als Related Articles | Metrics

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Effects of immunosuppressive drugs on bone marrow progenitor cells differentiating into macrophages EM>in vitro/EM> 2010, 41 (4):  538-544.  doi: 10.3969/j.issn.0529-1356.2010.04.011 Abstract ( )   Objective To investigate the effects of immunospressive drugs supplemented in culture media which provided the microenvironment for the generation of marophages bone marrow progenitor cells (BMPCs) differentiating into macrophages. BR> Methods BMPCs were prepared asepticly from C57BL/6 mice killed by cervical dislocation and EM>in vitro/EM> cultured in the media containing macrophage colony-stimulating factor (M-CSF) and rapamycin (rapa), cyclosporin A (CsA) and paclitaxel respectively. The morphology and phenotype of induced macrophages from BMPCs were analyzed by microscopy and flow cytometry (FCM). Results Induced cells from BMPCs presented specific macrophage morphology. There were high expressions of F4/80 and CD11 in surface molecules of induced macrophages and a dramatically decreased expression of CD11c. However, compared with the control group, there was a significant decrease in counts of differentiating macrophages. There was no significant effect on cell cycle of induced macrophages from BMPCs treated with cyclosporin A and a significant effect on apoptosis of those treated with paclitaxel. BR> Conclusion Three immunosuppressive drugs used in the experiment may affect the development and differentiation of macrophages from BMPCs. BR> Related Articles | Metrics

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Effects of heroin dependence on rat’s epidermal growth factor, β-endrophin positive cells in testis and serum follicle-stimulating hormone 2010, 41 (4):  545-549.  doi: 10.3969/j.issn.0529-1356.2010.04.012 Abstract ( )   Objective To explore the effects of heroin dependence on the epidermal growth factor(EGF), β-endrophin(β-EP) positive cells in the testis of rats and the changes of the serum levels of follicle-stimulating hormone (FSH), comprehend the possible functions of EGF, β-EP and FSH during heroin dependence. Methods Immunohistochemical SABC method, radioimmunoassay (RIA), counting the number of cells and image analysis were used in the studies. Results 1. Compared with the normal control group (NCG) and saline control group (SCG), the number of EGF positive cells decreased ( P<0.05 ), the intensity of staining stepped up gradually with the passage of time during dependence. The result of image analysis showed that the mean gray degree of the EGF positive cells decreased. 2. The number of β-EP positive cells in heroin dependence group(HDG) were less than that in NCG and SCG ( EM>P/EM><0.05 ).The immunohistochemical staining intensity of β-EP positive cells in testis decreased as compared with SCG and NCG and the means of gray degree increased gradually during heroin dependence (EM>P/EM><0.05). 3. The RIA result showed that the level of FSH was the lowest on the 10th day and then elevated as compared with SCG and NCG on the 17th day during heroin dependence. Conclusion During heroin dependence, the secretion function of the testis EGF positive cells was enhanced while the function the β-EP positive cells was weakened. The number of EGF,β-EP positive cells decreased markedly in testis. The secretory volume of FSH changes. The above findings indicated that testis ma Related Articles | Metrics

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Insulin promotes the proliferation of rabbit limbal stem cells EM>in vitro/EM> 2010, 41 (4):  550-553.  doi: 10.3969/j.issn.0529-1356.2010.04.013 Abstract ( )   Objective To get limbal stem cells(LSCs)by primary culture and explore the influence of insulin on LSCs’proliferation. Methods The cells from the rabbit corneal limbal tissue were cultured and identified by immunohistochemical staining and transmission electron microscopy. The LSCs were treated with 5mg/L, 10mg/L, 20mg/L, 40mg/L, 80mg/L and 100mg/L insulin respectively. Then methyl thiazolyl tetrazolium(MTT)assay was used to explore the influence of insulin on LSCs’ proliferation. Results The experiment groups at the concentration of 5mg / L and 10mg / L had significantly higher cell viability than that of the control group（EM>P/EM>0.05）, and so were the 5mg/L group and 10mg/L group（EM>P/EM>0.05）. But there were no significant difference between the experimental groups at th Related Articles | Metrics

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Influence of wheat germ agglutinin on cell growth and expressions of apoptosis genes in human breast cancer MDA-MB-231 cells 2010, 41 (4):  554-558.  doi: 10.3969/j.issn.0529-1356.2010.04.014 Abstract ( )   Objective To study the influence of wheat germ agglutinin (WGA) on cell growth and expressions of apoptosis genes in MDA-MB-231 cells. Methods MDA-MB-231 cells were cultured in DMEM containing 10% fetal bovine serum at 37℃ in a humidified atmosphere with 5% COSUB>2/SUB>.The experiment was performed on five groups: WGA groups were given 0.035, 0.07,0.14,0.28μmol/L WGA respectively, while in the control group the cells were merely grown in DMEM medium solution. Acid phosphatase assay(APA),flow cytometry（FCM） and Western blotting were used to detect the changes of cell viability,apoptosis ratio,mitochondrial transmembrane potential, the expressions of apoptosis-related protein poly ADP-ribose polymerase(PARP) and cytochrome C. Each experiment was done four times independently. Results Cell growth was inhibited after MDA-MB-231 cells were co-cultured with different concentrations of WGA for 24, 48, and 72 hours. Compared with human breast epithelia cells line HBL-100, MDA-MB-231 cells were more sensitive to WGA. After cells were treated with WGA for 24 hours, cell numbers reduced, volumes diminished.Early apoptosis ratio caused by WGA(0.14μmol/L, 0.28μmol/L,24 hours) were ( 28.7±3.48)% and ( 30.15±3.96)%, while the late apoptosis rates were ( 53.66±4.21)% and ( 63.18±5.53)%. Mean fluorescence intensity (MFI) of mitochondrial transmembrane potentials in MDA-MB-231 cells decreased.Apoptosis-related protein PARP showed an cleavage and cytochrome C expression increased in a dose-depandance manner. Conclusion WGA could significally restrain cell growth of breast cancer cell line Related Articles | Metrics

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Detection of DNMT3b and HDAC1 binding to SLC22A18 gene promoter by optimised chromatin immunoprecipitation 2010, 41 (4):  559-564.  doi: 10.3969/j.issn.0529-1356.2010.04.015 Abstract ( )   Objective To establish effective chromatin immnuoprecipitation (ChIP) and identify the interaction between the chromatin DNA of MDA-MB-231 cells and DNA methyltransferase 3b (DNMT3b), histone deacetylase 1(HDAC1) protein by ChIP techique. Methods The condition of ChIP assay was established and optimized. The MDA-MB-231 cells were treated with either 5-aza-2′-deoxycytidine (5-aza-dc), trichostatin A (TSA) seperately or the combination of the two drugs. Then DNMT3b and HDAC1 binding to SLC22A18 gene promoter region were detected by ChIP assay using specific DNMT3b and HDAC1 antibodies. PCR amplified its gene specific DNA fragemnt. Western blotting was used to examine protein levels of DNMT3b and HDAC1. Results SLC22A18 gene specific fragments were detected in the DNA fragments immunoprecipitated by DNMT3b and HDAC1 antibodies. DNMT3b was dissociated from the SLC22A18 promoter after treatment of MDA-MB-231 cells with 5-aza-dc alone or in combination with TSA. Treatment of MDA-MB-231 cells with TSA or in combination with 5-aza-dc reduced the association of HDAC1 with SLC22A18 promoter. The DNMT3b and HDAC1 protein levels were unaffected by either 5-aza-dc, TSA alone or combined together. Conclusion The results confirm that DNMT3b and HDAC1 was binded to SLC22A18 promoter in MDA-MB-231 cells and were involved in transcriptional regulation of SLC22A18 gene. Related Articles | Metrics

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Developmental changes of endocrine cells in the digestive tract of EM>Bufo gargarizans/EM> during metamorphosis 2010, 41 (4):  565-571.  doi: 10.3969/j.issn.0529-1356.2010.04.016 Abstract ( )   Objective To study the developmental changes in distributional density, histological localization and morphology of five types of endocrine cells in the digestive tract of EM>Bufo gargarizans/EM> during metamorphosis with gut hormone antisera of 5-hydroxytryptamine (5-HT), gastrin (GAS), somatostatin (SS), glucagon (GLU) and pancreatic polypeptide (PP). Methods Six samples were used at every developmental stage. The endocrine cells of digestive tract were studied by immunohistochemical techniques of streptavidin peroxidase (SP) method. Results 5-HT cells were detected in the digestive tract at every stage of EM>Bufo gargarizans/EM> during metamorphosis, and they first occurred in the middle portion of the digestive tract at hatching stage. They were distributed in the whole digestive tract after prometamorphosis stage, and the density of these cells in the middle portion of the digestive tract increased significantly (EM>P/EM><0.05). The density of 5-HT cells significantly increased in the esophagus and cardia from metamorphosis climax stage to juvenile stage ( EM>P /EM><0.05); GAS cells first occurred at juvenile stage, and they were mainly distributed in the duodenum and occasionally detected in the pylorus; SS cells first appeared at metamorphosis climax stage and they were occasionally distributed in the fundus and duodenum. SS cells were distributed in every portion from the esophagus to the jejunum at juvenile stage, and the density of these cells significantly increased (P<0.05); GLU cells were detected in the digestive tract at every stage of Bufo gargarizans during metamorphosis and the distributional density of GLU cells in the middle portion of the digestive tract significantly increased (P<0.05) after prometamorphosis stage; PP cells were not detected at every stage of Bufo gargarizans during metamorphosis. At hatching stage，the endocrine cells were located in the lamina propria with round in shape; At premetamorphosis stage and prometamorphosis stage, they were located in the epithelium of the whole digestive tract with round, oval or shuttle in shape; At metamorphosis climax stage, they were located in the gastric gland or the basal portion of epithelium of mucosa of the digestive tract with oval or taper in shape and occasionally detected in the lamina propria; At juvenile stage, they were located in the gastric gland and the epithelium of mucosa of the digestive tract with round or oval in shape, and some cell had a long cytoplasmic process reaching into the digestive lumen. Conclusion Comparing with other amphibians, there are both common and specific features in the developmental changes of endocrine cells in the digestive tract of Bufo gargarizans during metamorphosis, and the developmental changes of endocrine cells are adaptive to the changes of the digestive physiological activities and the individual physiological functions. Related Articles | Metrics

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Influence of annexin A7 low expression on proliferation and apoptosis of HeLa cells 2010, 41 (4):  572-575.  doi: 10.3969/j.issn.0529-1356.2010.04.017 Abstract ( )   Objective To study whether annexin A7(ANXA7) low expression can affect apoptosis and proliferation of HeLa cells. Methods Western blotting was used to identify whether siRNA can highly surpress annexin A7 expressin. When the effect of the siRNA was assured, the siRNA was transfected into HeLa cells by lipofectamine 2000. At the same time, cells in negative control group were transfected negative siRNA while cells as blank control group did not receive any treatment. After transfection for 48 hours, resazurin assay was conducted and DNA content was analyzed by cytometry. Results Resazurin assay showed that proliferation of the siRNA interference cell group decreased significantly（EM>P/EM><0.05）. Cytometry test showed that apoptosis of the siRNA interference group increased obviously（EM>P/EM><0.05）. Conclusion ANXA7 low expression may a Related Articles | Metrics

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EM>In vivo/EM> expression at early stage of bone growth factors and osteocalcin on the magnesium alloy/bone interface in New Zealand white rabbits 2010, 41 (4):  576-580.  doi: 10.3969/j.issn.0529-1356.2010.04.018 Abstract ( )   Objective To investigate EM>in vivo/EM> expression at early stage of three bone growth factors and osteocalcin on magnesium alloy/bone interface in New Zealand white rabbits. Methods The Ca-P coated and the naked Mg-Mn-Zn alloy rods were implanted respectivelys into the left femoral metaphysis of rabbits. Intravital staining was performed using subcutaneous injections of 1% water solution of calcein to observe the newly formed bone. Postoperatively, three rabbits were sacrificed at 1, 2, 3 and 4 weeks for X-ray examination, fluorescent observation, routine pathological examination and bone morphogenetic protein （BMP）, transforming growth factor βSUB>1/SUB>（TGF-βSUB>1/SUB>）, platelet-derived growth factor （PDGF） and bone gla protein（ BGP） immunohistochemistry. Results X-ray examination showed that both experimental and controlled rods were encircled by new bone, and HE staining and fluorescent observation showed no differences between two groups, but immunohistochemistry and associated statistical ananlysis displayed obvious differences on average absorbance values of BMP-2, TGF-β1 and PDGF expression. Average absorbance values of experimental group expressed higher than thoese of cont Related Articles | Metrics

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P>Expression of trefoil factor family-2 in gastric mucosa during healing of experimental gastric ulcer in rats/P> 2010, 41 (4):  581-586.  doi: 10.3969/j.issn.0529-1356.2010.04.019 Abstract ( )   Objective To explore the expression of trefoil factor family2(TFF2) in gastric mucosa during healing of experimental gastric ulcer of rats. Methods Acetic acidinduced ulcers were produced by injection acetic acid to submucosa of gastric anterior wall. The expressions of TFF2 peptide/mRNA in gastric ulcer group(GU,42 adult male rats), normal salinel group(NS,42 adult male rats,) and normal group(N,6 adult male rats) were detected by immunohistochemistry (IHC) and RT-PCR respectively. Results The results of IHC showed that the expression of TFF2 peptide in N group was weakly positive, and on the day 1 after acetic acid injection, TFF2 positive cells significantly increased,and reached the peak on the day 6TFF2 showed more stronger expression near muscularis mucosa,which mainly located in the cytoplasm of parietal cells. On the day 10 and 14 after acetic acid injection, the integral absorbance(IEM>A/EM>) values of TFF2 were lower than that on the day 6, but maintained a higher expression.On the day 23,the expression of TFF2 in GU group significantly decreased,but was more than that in the control groups(EM>P/EM><0.05 or EM>P/EM><0.01).The result of RTPCR showed that the change of TFF2 mRNA was consistent with TFF2 peptide,TFF2 mRNA showed more stronger expression on the day 4 and 6 in GU group. Conclusion TFF2 plays Related Articles | Metrics

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Changes and their significance of C3a, IFN-γ and leukemia inhibitory factor expressions in the decidua of rats vaginally infected with chlamydia trachomat 2010, 41 (4):  587-591.  doi: 10.3969/j.issn.0529-1356.2010.04.020 Abstract ( )   Objective To investigate the changes of C3a, interferonγ（IFN-γ）and leukemia inhibitory factor（LIF） expressions during the embryo implantation phase in decidua of rats vaginally infected with chlamydia trachomat（CT）. Methods Thirty adult female SD rats were randomly divided into experiment group and control group. Rats of the experiment group were vaginally infected with chlamydia trachomatis serotyped D and those of control group were vaginally injected with identical volume of isotonic sodium chloride. In both groups,rats were killed on the 5th,6th and 7th day of pregnancy. C3a, IFN-γ and LIF were detected in the decidua by immunohistochemical staining. Results C3a, IFN-γand LIF expression were detected in all of the 30 rats. C3a and IFN-γ expressions in the experiment group were both lower than those of the control group(EM>P/EM><0.01); On the 5th and the 6th days, LIF expression in the experiment groups was lower than that of the control group(EM>P/EM><0.01), and on the 7th day there was no difference between the two groups(P>0.05). In the experiment group, the expressions of IFN-γ and C3a were relevant positively（EM>r/EM> Related Articles | Metrics

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Effects of 5-HTSUB>4/SUB> receptor regulator on 5-HT release in gastrointestinal tract and liver regeneration in rats after partial hepatectomy 2010, 41 (4):  592-597.  doi: 10.3969/j.issn.0529-1356.2010.04.021 Abstract ( )   Objective To investigate the relationship of 5-HT in gastrointestinal tract with liver regeneration using 5-HTSUB>4/SUB> receptor regulator in rats after partial hepatectomy (PH). Methods Adult SD rats (n = 60) with PH were divided into three groups: control, cisapride and GR113808 group. After PH, Cisapride (a 5-HTSUB>4/SUB> receptor agonist, 10 mg/kg, twice per day, ig) and GR113808 (a 5-HTSUB>4/SUB> receptor antagonist, 3 mg/kg, twice per day, ip) were administered to the corresponding groups, respectively. Four kind of tissues (blood, stomach, small intestine and liver) were taken out at 0 hour, 24 hours, 48 hours, 72 hours, respectively, then the ratios of liver mass to body weight was accounted. 5-HT immunoreactive cells (5-HTIR cells) in stomach and small intestine were detected by immunohistochemical technique. The average gray level of 5-HTIR cells was detected by image analysis system. The plasma 5-HT level was determined by enzyme-linked immunosorbent assary(ELESA). The numbers of argyrophil nucleolar organizer regions(AgNORs) were detected by silver staining technique. Results Compared with the control, 1. The amounts of 5-HTIR cells in both stomach and small intestine decreased at 48-72 hours (EM>P/EM><0.05), the gray level of 5-HTIR cells increased significantly at 24-72 hours when plasma 5-HT level increased significantly(P<0.05), both the ratios of liver mass to body weight and AgNORs particles numbers in liver increased significantly at 48-72 hours(P<0.05), in cisapride rats with PH; 2. The amounts of 5-HTIR cells in both stomach and small intestine had not statistical difference at 24-72 hours when both the gray level of 5-HTIR cells and plasma 5-HT level decreased significantly (EM>P/EM><0.05 or EM>P/EM> <0.01), the ratios of liver mass to body weight decreased significantly at 4872 hours, and AgNORs particle numbers in liver decreased significantly at 24-72 hours (EM>P/EM><0.05 or EM>P /EM> <0.01), in GR113808 rats with PH. Conclusion Secretion of 5-HT in gastrointestinal tract was changed by 5-HTSUB>4/SUB> receptor regulator, which resulted in the changes of both the ratios of liver mass to body weight and the transcriptional activity of regenerating hepatocytes. In conclusion, 5-HT from gastrointestinal tract can significantly promote hepatocyte proliferation after partial hepatectomy. Related Articles | Metrics

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Expression and localization of Smad2 and Smad4 in domestic cat testis during testicular development and spermatogenesis 2010, 41 (4):  598-602.  doi: 10.3969/j.issn.0529-1356.2010.04.022 Abstract ( )   Objective To investigate the expression and localization of downstream signaling molecules of transforming growth factor beta (TGF-β) superfamily, Smad2 and Smad4 proteins, in postnatal cat testis, and to explore the function of Smad2 and Smad4 proteins in the regulation of testicular development and spermatogenesis. Methods Cellular localization of Smad2 and Smad4 proteins in juvenile, pubertal and sex maturity testes were examined by immunohistochemistry. Western blotting was performed to confirm that the anti-Smad2 and Smad4 antibodies would recognize specific proteins of the appropriate molecular weight in cat testis. BR> Results Smad2 and Smad4 mainly localizated in germ cells, Sertoli cells and Leydig cells of cat testis. The polyclone rabbit anti-Smad2 and Smad4 antibodies recognized a single band of about 58kD and 66kD in the cat testes, respectively. Conclusion Smad2 and Smad4 proteins are expressed in testicular cells during testicular development and spermatogenesis. It suggests that these two signaling molecules may play an important role in the regulation of cat testicular development and spermatogenesis. BR> Related Articles | Metrics

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Expression of annexin A7 in human uterine cervical squamous carcinomas and normal tissues 2010, 41 (4):  603-605.  doi: 10.3969/j.issn.0529-1356.2010.04.023 Abstract ( )   Objective To detect the expression of annexin A7 in human uterine cervical squamous cell carcinoma tissues and normal tissues. Methods SP immunohistochemistry was used to observe the differential expression of annexin A7 between carcinoma and normal paraffin imbedded specimen, each 25 cases. Western blotting and RT-PCR were used to observe the differential expression of annexin A7 between human uterine cervical squamous cell carcinoma and normal fresh tissues,each 25 cases. Results The annexin A7 expression in the tumor tissues was much higher than that of the normal tissues at the protein and the mRNA levels (P<0.05). Conclusion The expression of annexin A7 is upregulated in human uterine cervical squamous cell carcinoma tissues and it might be a new valuable tumor marker. Related Articles | Metrics

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P>Construction of highly organized three-dimensional muscle tissue induced by C2C12 cells EM>in vitro/EM>/P> 2010, 41 (4):  606-610.  doi: 10.3969/j.issn.0529-1356.2010.04.024 Abstract ( )   Objective To cultivate C2C12 cells on the modified moldgrooves of Sylgard 184 and induce C2C12 cells differentiating into highly organized threedimensional muscle tissue. Methods Bicomponents Sylgard 184 was mixed uniformly by the ratio of 10 ∶1 and poured into 6-well plates.Solidified by standing at room temperature, the surface of Sylgard 184 moldgrooves was casted and washed with Hank solution. Matrigel matrix and collagen were mixed uniformly, then paved at the bottom of grooves and airdried in the biological safety cabinet , making sure that the cell matrix material of coverage was uniform. After UV disinfection, C2C12 cells suspension was inoculated. When cells reached to 80% confluence,then C2C12 cells were differentiation cultivated. The morphology of myotubes was observed under the inverted microscopy. RT-PCR was used to test expression of myogenin and desmin mRNA in skeletal muscle tissue. Expression of myogenin and desmin muscle-specific protein was tested by immunofluorescence.Morphologies and inter-connections of myotubes were observed under scanning electronic microscope. Results As C2C12 cells were differentiation cultivated in the mold-grooves of Sylgard 184 for 7 days, differentiation polarity and myotube fusion were observed under the inverted microscope. After 21 days, it showed that myotubes stood closely and connected with each other detected by the scanning electron microscopy.The thickness of the membrane-like structure with the character of three-dimensional was 0.15mm.The positive expressions of myogenin and desmin mRNAs were detected in skeletal muscle tissue by RT-PCR. Immunofluorescence and DAPI nuclear staining showed that myogenin and desmin gene protein were expressed positively. Conclusion The modified mold-grooves of Sylgard 184 is able to guide the differentiation of C2C12 cells and promotes them to form parallel multinucleated myotubes . Myotubes could overlap with each other and form three-dimen Related Articles | Metrics

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Applied anatomy of the parameters of the iliac screw for lumbopelvic fixation 2010, 41 (4):  611-615.  doi: 10.3969/j.issn.0529-1356.2010.04.025 Abstract ( )   Objective To explore the entering spot, the length , the diameter and the direction of the iliac screw in the lumbopelvic fixation operations and provide anatomic basis for implanting correctly the iliac screw. Methods A total of 35 side hipbone specimens of native adults were studied.There were 30 side male specimens and 5 side female,16 left sides and 19 right sides.One line from the arc of the notch between posterior superior iliac spine and posterior inferior iliac spine was difined as the baseline(a).It was extended to acetabulum edge.Another line(b) was marked through the ischial spine and anterior superior iliac spine.It was intersected with baseline(a) in C spot. The C spot was located on the surface of ilium body behind the acetabulum and determinated.The fan-shaped lines were described in the posterior part of iliac through the C spot and the baseline.Five oblique sections were obtained through there fan-shaped lines. Various oblique sections were observed and correlation data above the 10°-30° oblique sections were measured.At the same time,the medial lip, lateral lip and intermediate line in the behind part of iliac were observed.After definiting the entering spot, the length, the diameter and the direction of the iliac screw,these data were verificated in 2 native adut cadavers . Results It was found that 35° oblique section arrivals to the iliac fossa and 5° oblique section clings to the arc of the greater sciatic notch. The diameter of the iliac screw based on f line which was the shortest distance between the gluteal surface and pelvic surface. The intermediate line on posterior part of the iliac spine could be distinguished clearly,which was like a regular arc and the most raised part. The minimum distances(K) from entering spots to the cartilage of the acetabulum roof was 109.38mm.  Conclusion Iliac screws can be implanted accurately and safely by the C spot,the baseline and the outer plate in 10°-30° posterior part of the iliac. The most entering spots are located in the points of the fanshaped lines intersecting with the Related Articles | Metrics

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Anatomical study on the relationship between the posterior sagittal septum of knee and the tibial insertion of posterior cruciate ligament 2010, 41 (4):  616-619.  doi: 10.3969/j.issn.0529-1356.2010.04.026 Abstract ( )   Objective to investigate the anatomical relationship between the posterior sagittal septum of knee and the tibial insertion of posterior cruciate ligament (PCL). Methods Twentytwo fresh frozen adult cadaveric knee joints were dissected. With the knee flexed to 90 degrees, PCL was divided into anterolateral bundle and posteromedial bundle. The outlines of the footprints of the two bundles were marked with ink. The posterior sagittal septum of knee and the insertion of PCL on tibia were observed and measured. Results There was a sagittal septum in the knee, the anterior of which was connected with fat pad, alar fold or mucous ligament. The middle part of the sagittal septum lied between the anterior and posterior cruciate ligament, the postetior part of the sagittal septum formed the posterior septum of the knee. Eight of the 22 knees (36.36%), the posterior sagittal septum passed the PCL at lateral side. And 14 of the 22 knees (63.64%), the sagittal septum passed around the bundle of PCL. Moreover, the center of the tibial insertion of anterolateral bundle of PCL located at （0.90±2.40）mm medially to the posterior sagittal septum ,and at （3.25±1.20）mm beneath the lateral tibial surface. The posteromedial bundle was seated at （4.35±2.46）mm medially to the posterior sagittal septum and at （6.91±1.57）mm beneath the lateral tibial surface. Conclusion The anatomical relationship between the posterior sa Related Articles | Metrics

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Clinical anatomic observation on the ureteropelvic junction 2010, 41 (4):  620-622.  doi: 10.3969/j.issn.0529-1356.2010.04.027 Abstract ( )   Objective To supply the clinical applied anatomic data for locating the ureteropelvic junction (UPJ). Methods The urinary system specimens of 32 fixed adult cadavers were used for general measurement and then, the middle part of renal pelvis(RPSUB>mid/SUB>), UPJ and the middle part of ureter(URSUB>mid/SUB>) were taken for HE and nevre fibers sliver staining and the histological features were observed. Meanwhile, the staining images of the three parts were analysed and measured by computer. Results The outer diameters of UPJ(3.12±0.81)mm, was among the shortest one of three outer diameters. Similarly, the inner diameter of UPJ (2.26±0.15)mm, was respectively shorter than that of RPSUB>mid/SUB> or URSUB>mid/SUB>(EM>P/EM><0.05) .The nerve fibers were less in UPJ and RPSUB>mid/SUB>,whereas, the collagen fibers were relatively richer in muscular fascicles of UPJ. The smooth muscles arranged irregularly in UPJ . Both the wall (0.63±0.04)mm and the muscular layer (0.34±0.03) mm of UPJ were thiner respectively than those of RPSUB>mid/SUB> and URSUB>mid/SUB>（EM>P/EM><0.05）. The average value of renal incline angle was 61°. Related the angle, the general position of UPJ was about 1.2-1.3 cm above the plane extended horizontally from renal inferior extremity (PSUB>inf/SUB>). The larger the incline angle(θ) was, the higher the UPJ located in. Conclusion The location of UPJ is generally about 1.25cm above PSUB>inf /SUB>.There are physiologic intracavitary narrow and co Related Articles | Metrics

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Effects of icariin on the level of nitric oxide and malondialdehyde during early embryo development of porcine 2010, 41 (4):  623-625.  doi: 10.3969/j.issn.0529-1356.2010.04.028 Abstract ( )   Objective To study the effects of icariin (ICA) on the level of nitric oxide (NO) and malondialdehyde (MDA) in culture medium of fertilization(IVF)early porcine embryos in vitro, and to research both the relations among ICA, NO, MDA and the association with developmental ability of porcine IVF embryos. Methods NCSU-23 basic culture medium was used as the medium for control group, while ICA(0.6mg/L)supplemented medium was used as the medium for the experiment group. The reductive ferment method and thiobarbituric acid colormetry method were used to detect the nitric acid and MDA in media at interval duration of culture (0-48h,48-120h,120-168h). Results There were significantly different between experimental (342 embryos) and control (349 embryos) groups in the cleavage rate (74.84 vs 63.32%)(ICA vs control), the morrula and blastocyst rate (45.08 vs 34.10 %), and the blastocyst rate(23.38 vs 18.04%) at the intervals for 48 th, 120 th,168 th hour(EM>P/EM><0.01). During the culture process of porcine embryos, the concentrations for both NO and MDA in media kept increasing, while NO concentration in medium was higher in ICA group than that in the control group, the MDA concentration in medium was lower in ICA group than that in the control group, but there was no statistic significant(EM>P/EM>>0.05). 〖WTHZ〗Conclusion ICA can promote the developmental abilit Related Articles | Metrics

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Morphometry and their relationship of the main cutaneous arteries in rats 2010, 41 (4):  626-630.  doi: 10.3969/j.issn.0529-1356.2010.04.029 Abstract ( )   Objective To describe the morphometric and hemodynamic characteristics of main cutaneous arteries of rats. Methods The anatomy of cutaneous arteries was studied using a modified lead oxidegelatin injection technique in 12 rats. Angiograms were assembled with Adobe Photoshop and analyzed with Scion Image Beta to determine the scope of blood supply, the trend in blood vessel density and blood supply. Results The main source vessels contributed a summed mean of 6 cutaneous arteries in the rat trunk, with a mean emerging vessel diameter of (0.53±0.12)mm，the perimeter of the skin that could potentially be supplied by the cutaneous artery averaged (18.74±4.84)cm, and its mean vascular density was (391.31±76.58) gray value/(pixelI>&#/I>8226;mm）. It was positive correlation between the diameter and its supplying perimeter, the linear regression was y=32.441x+1.233( EM> r/EM> =0.851，EM> n/EM> =54,EM>P/EM><0.01). Conclusion The diameter and the perimeter of its nutrient region is directly proportional. In the case of the same perimeter, nutritional distribution is more close to the regional round, the greater the area of nutrition. The Related Articles | Metrics

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Photoconvertion of DiI and the preparation of the electron microscope sample 2010, 41 (4):  631-633.  doi: 10.3969/j.issn.0529-1356.2010.04.030 Abstract ( )   Objective In order to make the labeled neurons and glia with 1,1′-dioctadecy-3,3,3′,3,-tetramethylindocarbocyanine perchlorate(DiI) diolistic labeling for long time preservation and ultrastructural study. Methods DiI diotistic assays were done on the sections of visual cortex of 20 C57/B6J mice to label neurons and glia, then the targeted neurons were photoconverted and prepared for ultrathin section. Results The neurons after the photoconversion could still show fine shape with spines on dendrites and decent synapse ultrastructure. Conclusion After DiI diotistic labeling and photoconversation, the labeled neurons can present satisfied morphological details with either light microscopy or electron microscopy. The organelle is well and the synaptic structure is clear either. Related Articles | Metrics