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2010, Volume 41 Issue 5
06 October 2010 Previous Issue    Next Issue
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Response of the capillaries and astrocytes in the spinal cord dorsal horn of rats induced by spared nerve injury 2010, 41 (5):  635-640.  doi: 10.3969/j.issn.0529-1356.2010.05.001 Abstract ( )   Objective To investigate the response of the capillaries and astrocytes in the spinal cord dorsal horn of rats after the spared nerve injury (SNI). Methods Male SD rats were randomly divided into two groups and used for light microscopy(n=30) or electromicroscopy(n=10) study respectively. The rats for light microscopy study were divided into control, sham-SNI and SNI subgroups. Ten rats were performed SNI operation and survived for 20d respectively. Five of them were injected Evans blue into caudal vein, 30min before perfusion, the L4 segment of spinal cord was removed and cut into 30 μm frontal sections on a cryostat. The section was detected under Olympus BX51 fluorescent microscope. Other five of them were perfused, the sections were performed anti-rat IgG and glial fibrillary acidic protein (GFAP, a marker for astrocyte) immunohistochemical staining (ABC method). Rats used for immunoelectromicroscopy were divided into control and SNI 20 days subgroups, the spinal cord sections were performed anti-rat IgG immuno-electromocroscopic staining. Results 1. On 20 days after SNI, many clear red effusion spots of Evans blue were observed in the dorsal horn. 2. SNI induced astrocytes became active type and the number were increased; IgG positive granules on the capillary wall and perivascular space were increased markedly.3. Immunoelectromicroscopy showed that at 20 days after SNI, IgG positive granules were increased, the following changes of the endothelial cells were found: (1)Many microprocesses extended from the endothelial cells were observed, some of them contained IgG positive granules. (2)Many “endocytosis concave” on the luminal side, vesicles in cytoplasm and “exocytosis steps” on aluminal side of endothelial cells containing IgG positive granules were observed respectively. （3）The tight junction of endothelial cells revealed gap and containing IgG positive granules.(4) Many “vesicles” revealed within the cytoplasm, some of them contained IgG positive granules and some did not. Conclusion After SNI the Related Articles | Metrics

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Effects of X-ray on structure of cerebral occipital lobe cortex and activities of superoxide dismutase, catalase and content of malonaldehyde in cerebrum of filial mice 2010, 41 (5):  641-648.  doi: 10.3969/j.issn.0529-1356.2010.05.002 Abstract ( )   Objective To investigate the dynamic changes of structure of cerebral occipital lobe cortex and cerebral weight and activities of superoxide dismutase(SOD),catalase(CAT) and content of malonaldehyde(MDA) after irradiation with different dosages of X-ray in filial mice,and to explore effects of X-ray on cerebral development of filial mice. Methods One hundred and sixty filial mice（birth 6-7 days）were irradiated with different dosages (0Gy,1Gy,3Gy,5Gy,7Gy) of X-ray ,At 1day, 5days, 10days and 20days after irradiation,cerebral weight of filial mice was weighted. And it was observed that the changes of the structure of the cerebral occipital lobe cortex of filial mice by biomicroscopy and detected the activities of SOD, CAT and content of MDA by colorimetry from all irradiated groups of different stages. Results X-ray affected the development of cerebrum in filial mice , at 1 day-20 days after irradiated,cerebral weight of other irradiated groups was lighter than that of the control group except 1Gy irradiated group(EM>P/EM><0.05 or EM>P/EM><0.01);SOD and CAT activities increased and MDA content decreased in 1Gy group at 5-20 days after irradiation; In 3Gy group,SOD and CAT activities decreased firstly and then increased slowly, but it was always lower than those of control group and MDA content increased in the early time, then decreased, but it was always higher than that of the control group; SOD and CAT activities in 5Gy,7Gy group filial mice were significantly lower than that of the control group(EM>P/EM><0.01), and MDA content significantly was higher than that of the control group(EM>P/EM><0.01);The thickness of cerebral occipital lobe cortex in irradiated groups decreased, structure of cerebral occipital lobe cortex was not clear, the number of neurons was lower than that of the control group except those in cerebral occipital lobe cortex II,III,VI at one day and those in cerebral occipital lobe cortex III,V,VI at 20 days after irradiation in 1Gy group filial mice. Conclusion X-ray radiation affects the cerebral weight and the structure of filial mouse cerebrum, this effect might be correlated with the high activities of SOD, CAT and low content of MDA in filial mouse brain. Related Articles | Metrics

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Effects of ghrelin, melanotan-II and cholecystokinin on food intake in rats with paraventricular nucleus lesions 2010, 41 (5):  649-653.  doi: 10.3969/j.issn.0529-1356.2010.05.003 Abstract ( )   Objective To investigate the mechanism of hyperphagia and obesity induced by bilateral lesions of the hypothalamic paraventricular nuclei (PVN). Methods Wistar male rats (36 rats) were used. After bilateral electrolytic lesions of the PVN, we measured the effects of ghrelin, melanotan-II (MT-II, a synthetic structural homologue of alpha-melanocyte-stimulating hormone, α-MSH) and cholecystokinin8 (CCK-8) on food intake in rats with PVN lesions. Results After bilateral electrolytic lesions of the PVN, the body weight and food intake were significantly increased, showing hyperphagia and obesity. After 1 week of bilateral electrolytic lesions of the PVN, the rats were given ghrelin and CCK-8 intraperitoneally, or intracerebroventricular infusion of MT-II. We measured the cumulative food intake (FI) for 4 hours after ghrelin injection in rats fed ad lib, and the changes in FI at 15min, 30min, 1 hour and 2 hours after administration of MT-II and CCK-8 in rats fasted for 24 hours. Ghrelin significantly increased cumulative FI, with maximal response 3 hours and 4 hours after injection, and at these times, the FI of PVN-lesioned rats was greater than that of sham-operated rats. MT-II decreased FI in shamoperated, but not in PVNlesioned rats. CCK-8 decreased FI at each time point in all groups, there was no difference between PVN-lesioned and sham-operated rats. Conclusion The hyperphagia and obesity induced by PVN lesions might be related to an increased orexigenic action of ghrelin d Related Articles | Metrics

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Periventricular white matter change and matrix metalloproteinase-2 and 7 expression after chronic cerebral hypoperfusion in rats 2010, 41 (5):  654-659.  doi: 10.3969/j.issn.0529-1356.2010.05.004 Abstract ( )   Objective To investigate periventricular white matter change and matrix metalloproteinase-2 and 7（MMP-2 and MMP-7） expression after chronic cerebral hypoperfusion in rats. Methods Sixty healthy male Wistar rats were randomly divided into sham-operated (control) group and three model groups(20 days, 40 days and 60 days). Model of chronic cerebral hypoperfusion was induced by staging permanent ligation of the bilateral common carotid arteries in the rat. The ultrastructure change in periventricular white matter was observed with transmission electron microscopy. The activation of astrocytes was assessed with immunohistochemistry staining of glial fibrillary acidic proteinl(GFAP), and the expressions of MMP-2, 7 and myelin basic protein(MBP) were detected with immunohisitochemisty staining and Western blotting. Results Irregular myelin sheaths and degenerating myelinated axon were observed at the ultrastructural level in the corpus callosum and internal capsule of the 20 days operated rats. Obvious proliferation hypertrophy of GFAP immunopositive astrocytes were observed and the expression of GFAP gradually increased in the model groups, while the content of MBP decreased（EM>P/EM>< 0. 01）. The expressions of MMP-2 and MMP-7 were markedly elevated in the model groups（EM>P/EM>< 0. 01 or EM>P/EM>< 0. 001）. Additionally, the up-regulation of MMP-2 and MMP-7 showed a significant relation with periventricular white matter matter lesions. Conclusion Chronic cerebral hypoperfusion induces progressive periventricular white matter lesions with the upregulation of MM Related Articles | Metrics

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Expression of intestinal trefoil factor on supraoptic nucleus and paraventricular nucleus in rat hypothalamus and its relationship with experimental gastric ulcer 2010, 41 (5):  660-665.  doi: 10.3969/j.issn.0529-1356.2010.05.005 Abstract ( )   Objective The purpose of this study is to explore the expression of intestinal trefoil factor (ITF) on supraoptic nucleus(SON) and paraventricular nucleus(PVN) in rat hypothalamus and its relationship with experimental gastric ulcer. Methods The changes of ITF in hypothalamus and blood serum in gastric ulcer group(GU) and normal group(N) rats were detected by means of immunohistochemical staining method and enzymelinked immunosorbent assay（ELISA).The transcription of ITF mRNA was detected by RT-PCR. Results ITF immunoreactive cells were mainly located in SON and magnocellular region of PVN(mPVN).The integral absorbance(IEM>A/EM>) was obviously increased from the day 2 to 6 after gastric ulcer(EM>P/EM> <0.01)，and reached a highest expression at the 6th day. then gradually decreased from 10 days to 23 days，but remained on a high level compared with the normal group(EM>P/EM><0.05). The change of ITF peptide in serum was similar to the immunohistochemical results. The absorbance of ITF/glyceral dehyde 3-phosphate dehydrogenase (GAPDH) was obviously higher in gastric ulcer group than that of the normal group from 2 days to 23 days（EM>P/EM><0.01 or EM>P/EM> <0.05）. Conclusion The high expression of ITF in rat hypothalamus and blood serum m Related Articles | Metrics

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Interferring effect and probable mechanism of neuregulin on experimental dementia model in rats 2010, 41 (5):  666-669.  doi: 10.3969/j.issn.0529-1356.2010.05.006 Abstract ( )   P>Objective To investigate the effect of neuregulin 1β (NRG1β) on the neuronal apoptosis and the expressions of Bcl-2 and Bax in experimental dementia model rats. Methods Thirty adult healthy male Wistar rats were randomly divided into control group, model group and treated group consisting of 10 rats respectively. The experimental dementia models were established by injecting beta-amyloid protein 140 (AβSUB>1-40/SUB>) stereotactically into the left lateral ventricle, and treated by injecting NRG1β into right lateral ventricle. The cognitive capacity of rats was evaluated with Y-electric maze. The neuronal apoptosis was counted by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL) assay. The expressions of Bcl-2 and Bax were determined with immunohistochemistry assay. Results The cognitive ability in model group rats decreased, along with the number of neuronal apoptosis and the expressions of Bcl-2 and Bax increased significantly than those in control group (EM>P/EM><0.05). After treatment with NRG1β, the cognitive ability of rats improved, the number of neuronal apoptosis reduced and the expression of Bcl-2 increased while Bax decreased significantly than those in model group (EM>P/EM><0.05). Conclusion NRG1β could inhibit neurona Related Articles | Metrics

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Influence of the nitric oxide synthase in the sympathetic nervous system on proliferation of vascular smooth muscle 2010, 41 (5):  670-673.  doi: 10.3969/j.issn.0529-1356.2010.05.007 Abstract ( )   Objective To investigate the influence of the nitric oxide synthase(NOS) in the sympathetic nervous system on proliferation of vascular smooth muscle. Methods The model of proliferation of vascular smooth muscle was established by injuring common carotid artery by FeClSUB>3/SUB>.Twenty-four SD rats were divided into 4 experimental groups：sham operation group, survival 1 day group，survival 5 days group and N-nitro-L-arginine(LNNA) treatment group. Cervical sympathetic ganglion and spinal cord intermediolateral nucleus were retrogradely labeled by injection of 3% fluorogold(FG) into carotid sheath and cervical sympathetic ganglion, and nicotinamide adenine dinucleotide phosphate diaphorase(NADPH-d) reactivity was detected histochemically. The proliferation of vascular smooth muscle was observed by hematoxylineosin(HE) staining. Results Compared to sham operation group， NADPH-positive neurons of cervical sympathetic ganglion and spinal cord intermediolateral nucleus were increased in survival 1day and 5 days group,especially 5 days group，but decreased in LNNA group.The proliferation of vascular smooth muscle was significantly increased in LNNA group as indicated by HE staining. Conclusion The NOS of the sympathetic nervous system plays an important role on proliferation of vascular smo Related Articles | Metrics

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NR1 Subunit of N-methy-D-asparate receptor regulates the radial migration of cortical neurons 2010, 41 (5):  674-678.  doi: 10.3969/j.issn.0529-1356.2010.05.008 Abstract ( )   Objective To investigate whether and how N\|methy\|D\|asparate(NMDA) receptor regulates radial migration of cortical neuron. Methods Migration route and characters of transefected neurons were traced, using EM>in utero/EM> electroporation of small interference RNA (siRNA) of NMDA receptor NR1 subunit, the essential subunit, and fluorescence immunohistochemistry. Results Cortical neurons transfected with control vector,could radially migrate to the proper cortical plate layer 2-3 before postnatal 7(P7), however, neurons which silenced NR1 subunit could not migrate to the proper layer 2-3, while improperly stayed at lower layers or even the germinal regions. Conclusion NMDA receptor plays an essential role in regulating proper neuronal migraion and cortical lamination in the developing rat cortex, which is helpful to understand the basic mechanism of neuron migration and prevent neuronal migration disorders. Related Articles | Metrics

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Expression and secretion of basic fibroblast growth factor in rat amniotic epithelial cells introduced by lentiviral vector 2010, 41 (5):  679-685.  doi: 10.3969/j.issn.0529-1356.2010.05.009 Abstract ( )   Objective To construct rat amniotic epithelial cells (AECs) modified with human basic fibroblast growth factor (bFGF) gene that combined with nerve growth factor (NGF) secreting peptide utilizing lentivirus vector, by which bFGF could be stably synthesized and secreted. Methods Amniotic epithelial cells were collected from the term rats by trypsin-treating process, and subcultured AECs were identified by immunocytochemistry and RT-PCR methods. Lentiviral vector with human bFGF gene was constructed, and lentivirus packaging was performed after DNA sequencing. Afterwards, AECs were infected with viral supernatant, then selected with blasticidin. Furthermore, the selected cells were detected for bFGF expression and secretion by immunocytochemistry, RT-PCR and ELISA methods. Thereafter，growth activity of AECs chimaera was investigated, and the biological functions of secreted bFGF was evaluated by culturing PC12 cells with conditioned medium of AECs chimaera. Results Immunocytochemical staining and RT-PCR showed that the rat AECs expressed epithelialspecific markers CK-19, neural cell markers nestin and GFAP, as well as pluripotent cells markers SSEA-4, Oct-4, Nanog, Sox2 and vimentin. The AECs chimaera sustainable expressing bFGF were obtained after infection and screening processes. Immunocytochemistry staining and RT-PCR showed that most of the AECs chimaera expressed bFGF, compared with the nonexpression before transfection. ELISA results implicated that AECs chimaera were able to secrete bFGF peptide. AECs chimaera survived in the screening procedure grew much faster compared with common AECs. The conditional culture medium of AECs chimaera could stimulate PC12 cells growth and neurites development, as compared with AECs group. Conclusion bFGF gene could be transferred into rat AECs successfully by lentiviral vector, and the AECs chimaera could express and secret bFGF afterwards, indicating robust growth activity and significant neurotroph Related Articles | Metrics

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Establishment of an aging model of Sca-1SUP>+/SUP> hematopoietic stem cell and its related biological mechanism 2010, 41 (5):  686-692.  doi: 10.3969/j.issn.0529-1356.2010.05.010 Abstract ( )   Objective To establish the aging model of hematopoietic stem cells(HSCs) and investigate its related biological mechanism. The purpose is to build the foundation for searching the methods of delaying HSCs aging. Methods Sca-1SUP>+/SUP>HSCs were isolated and purified by magnetic activated cell sorting (MACS). The purity of separated cells was analysed by flow cytometry (FCM) and the expression of Sca-1SUP>+/SUP> antigen was detected by immunofluorescence. Sca-1SUP>+/SUP>HSCs were induced by tertbutylhydroperoxide( t-BHP, final concentration of 100 μmol/L) for 6 hours to establish the HSCs aging model EM>in vitro/EM>. Biological characteristics of aging HSCs was evaluated by mixed hematopoietic progenitor cell culture, cell cycle assay and sensscence-associated β-galactosidase (SA-β-gal) cytochemical staining. Telomere length and telomerase activity were detected by Southern blotting and TRAP-PCRSYBR Green staining. By using reverse transcription polymerase chain reaction(RT-PCR), the expression of p16SUP>Ink4a/SUP>, p19SUP>Arf/SUP>, p53, p21SUP>Cip1/Waf1/SUP>mRNA was detected. Results The purity of separated Sca-1SUP>+ /SUP>HSCs was （87.33±1.25）%. After being cultured with 100 μmol/L t-BHP for six hours, the ability of aging Sca-1SUP>+/SUP> HSCs to forming mixed hematopoietic progenitor colony, self-renewal and multidifferentiation was decreased significantly. The number of aging Sca-1SUP>+/SUP> HSCs entered GSUB>1/SUB> phase of the cell cycle, the percentage of SA-β-gal positive cells and the expression of p16SUP>Ink4a/SUP>, p19SUP>Arf/SUP>, p53, p21SUP>Cip1/Waf1/SUP>mRNA were increased. The telomere length was shorten and the telomerase activity was Related Articles | Metrics

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Effects of Laminaria japonica polysaccharide to reduce serum lipidsand anti-inflammation in hyperlipenmia rats 2010, 41 (5):  693-697.  doi: 10.3969/j.issn.0529-1356.2010.05.011 Abstract ( )   Objective To investigate the regulating effects and mechanism of Laminaria japonica polysaccharide (LJPS) on lipid levels in serum of hyperlipenmia rats. Methods Thirtytwo healthy female Wistar rats were used to establish hyperlipemia models by feeding fatrich forage, and treated with LJPS as a potential drug. The serum levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL) and high-density lipoprotein (HDL) were determined by biochemical assay. The nitric oxide (NO) level in serum and hepatic tissue was measured by nitrate reductase assay and the activity of superoxide dismutase (SOD) in serum and hepatic tissue was detected by oxidase assay. The level of ox-LDL in hepatic tissue was determined by enzyme-linked immunosorbent assay. The expressions of SOD and inducible nitric oxide synthase (iNOS) in hepatocytes were detected by immunohistochemical assay. Results The rat serum levels of TG, TC and LDL were significantly higher while HDL was lower in the LJPStreated group than those in the hyperlipidemia model group (EM>P/EM><0.05). Both the levels of ox-LDL and NO in both the rat serum and hepatocytic tissues were significantly lower and the activity of SOD was higher in the LJPS-treated group than those in the hyperlipidemia group (EM>P/EM><0.05). The experimental results showed that the iNOS down-expression and the SOD upexpression were found in the LJPS-treated group than those in the hyperlipidemia group (EM>P/EM><0.05). Conclusion Laminaria japonica polysacharide- mi Related Articles | Metrics

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Genetic instability and methylation of bone morphogenic protein 3 gene in the gastric cancer in Chinese 2010, 41 (5):  698-701.  doi: 10.3969/j.issn.0529-1356.2010.05.012 Abstract ( )   Objective This project is to explore the influence of microsatellite instability (MSI), loss of heterozygosity (LOH), as well as promoter region methylation in bone morphogenetic protein 3 (BMP3) on gastric carcinoma development. Methods LOH and MSI of locus D4S2922 and D4S2964 in 50 primary gastric carcinoma specimens were detected by polymerase chain reaction\|single strand conformation polymorphism analysis (PCR\|SSCP) and methylation-specific PCR analysis in the study. Results The positive rates of MSI, LOH was 16.00% and 20.00% in 50 specimens respectively, and the frequency of LOH in lymph node metastasis cases was significantly higher than those without. TNM stage III and IV also exhibited higher LOH frequency compared with stage I and II. The positive rate of methylation on BMP3 gene promoter was 64.44% in 45 specimens. Moreover, the MSI negative group exhibited higher methylation frequency than that of MSI positive group. Conclusion These data suggested that BMP3 genetic instability and promoter methylation were initiated during the gastric carcinogenesis. LOH was detected mostly in the late stage of the gastric carcinoma progression, which indicated higher carcinoma infiltration and poor prognosis. Promotor region methylation of BMP3 gene might be causative in the gastric carcinoma cases in Chinese. Related Articles | Metrics

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TGF-β1 stimulates Snail production in rat bone mesenchymal stem cells through activation of ERK and PI3K pathways 2010, 41 (5):  702-708.  doi: 10.3969/j.issn.0529-1356.2010.05.013 Abstract ( )   Objective To examine whether transforming growth factor\|β1(TGF-β1) could induce expression of Snail in bone marrow mesenchy mal stem cells (BMSCs) through activation of extracellular signal\|regulater kinases(ERK) and PI3K. Methods Density gradient centrifugalization combined with adherence method was used to segregate rat BMSCs.The BMSCs were cultivated to the 3rd passage and characterized using flow cytometry technique.After BMSCs were treated with PD98059（0,15,30,50μmol/L）or Wortmannin(0,20,40,80 nmol/L)for 1hour，the levels of p-Akt and p-ERK1/2 with TGF-β1 treatment for 6hours were examined by Western blot to determine the effective concentrations for PD98059 and Wortmanin to inhibit the TGF-β1induced ERK and PI3K activity of cells.Then,we employed these specific inhibitors with respective effective concentrations to incubate BMSCs with TGF-β1 treatment for 6 hours or 24 hours and detect the expression of Snail mRNA and Snail protein by RT-PCR and Western blotting. Results Density gradient centrifugalization combined with adherence method could segregate and purify rat BMSCs effectively.The results of flow cytometry showed that CD29 and CD44 expression were positive while CD34 and CD45 expression was negative for BMSCs.After TGF-β1 treatment，increased levels of p-ERKs and p-Akt were confirmed by Western blotting.TGF-β1induced increased activity of p-Akt and p-ERKs in BMSCs could be inhibited in a concentrationdependent manner by Wortmannin and PD98059.The effective concentrations for PD98059 and Wortmanin to inhibit the TGF-β1induced ERK and PI3K activity of cells were 30μmol/L and 40nmol/L respectively.The significantly decreased levels of the Snail mRNA and Snail prot Related Articles | Metrics

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Associativity and mechanism of β-catenin with ovarian cancer metastasis 2010, 41 (5):  709-713.  doi: 10.3969/j.issn.0529-1356.2010.05.014 Abstract ( )   Objective To investigate associativity and mechanism of β-catenin with ovarian cancer metastasis. Methods The different expressions of β-catenin in high and low metastatic human ovarian serous cystadenocarcinoma cells（HO-8910PM and HO-8910） were detected by immunofluorescence staining, Western blotting and RT-PCR analysis. To further investigate the regulated mechanism of β-catenin with ovarian cancer metastasis, low metastatic human ovarian serous cystadenocarcinoma HO-8910 cells were treated by hepatocyte growth factor(HGF). The expression change of β-catenin in HO-8910 cells was investigated by the same technology mentioned above. Then, HO-8910 cells invasion and migration abilities were assessed by Transwell chambers and scratch wound migration assay. Results The expression of β-catenin was significantly lower in HO-8910PM cells than that in HO-8910 cells (P<0.05). HGF could downregulate the expression of β-catenin in HO-8910 cells (P<0.05) and enhance cell invasion (P<0.05) and migration. Conclusion The lower expression of β-catenin may be correlated with invasion and migration pot Related Articles | Metrics

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Effects of passive smoking on rat ovarian follicle-stimulating hormone, lutteinizing hormone and gonadotropin releasing hormone and its receptors 2010, 41 (5):  714-718.  doi: 10.3969/j.issn.0529-1356.2010.05.015 Abstract ( )   Objective To study the effects of passive smoking on Wistar rat ovarian structure, the expression of hormone receptor and serum hormone. Methods The experimental SD rats were treated with fired smoking cigarette for 3 months. The rats were sacrificed after 3 months treatment. Changes of rat ovarian structures were detected by light and electron microscope, the expression of hormone receptors was detected by immunohistochemistry, changes of serum hormone were detected by enzyme-linked immunosorbent assay. ResultsThe number of follicles and mature oocytes reduced, vascular contracted in medulla, and the mesenchymal loosed. Mitochondrial were fuzzy, water-like changed and showed vacuolization degeneration and cristae rupture in the granulosa cells of smoking rat. The rough endoplasmic reticulum was degranulated. The levels of follicle-stimulating hormone （FSH）, lutteinizing hormone （LH） and gonadotropin releasing hormone （GnRH） in sera were obviously lower in the smoking model group than that of the control group(EM>P/EM>0.01， EM>P/EM>0.05，EM>P/EM> 0.05). The intensity of FSHR and LHR in the ovarian tissues was significantly lower in the smoking model rats than that in the control rats for 3 months treatment(EM>P/EM>0.01，EM>P/EM>0.05). Conclusion Passive smoking can degrade the l Related Articles | Metrics

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Effect of age and genetic factor on mouse EM>in vitro/EM> fertilization 2010, 41 (5):  719-723.  doi: 10.3969/j.issn.0529-1356.2010.05.016 Abstract ( )   Objective Immature and mature mouse models were used and the effect of age on mouse EM>in vitro/EM> fertilization were identified in order to make full use of high quality animal resources, to shorten the intervals between generations and efficiently to improve the embryo engineering technology. Methods The ovulation rate, the in vitro fertilization rate and the blastocyst rate were measured after pregnant mare serum gonadotropin（PMSG）/ human chorionic gonadotrophin( hCG ) superovulation of both the immature and mature mice as well as the ovarian morphological changes were observed. At the same time, the role of genetic factor in the mouse in vitro fertilization was evaluated. Results The ovulation rate in immature mice was significantly higher than that in mature mice, and antral follicles in immature mouse ovaries were more than in mature mouse ovaries. There was a significantly lower fertilization rate from immature mouse oocytes than from mature mouse oocytes, however, there was no remarkable difference on the blastocyst rate between these two groups. Further analysis of usable oocytes in the mature mouse group found that the high response group to superovulation (usable oocytes ≥40) had significantly higher fertilization rate and blastocyst development rate than those of the low response group to superovulation (usable oocytes 40). In addition, the ovulation rate and the follicle development among different individuals in the same litter were similar. Conclusion There is a high response to superovulation in the immature mice, while in the mature group, the ovarian response to superovulation Related Articles | Metrics

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Relationship between the expression of trefoil factor 2 in rat submaxillary glands and the self healing of experimental gastric ulcer 2010, 41 (5):  724-727.  doi: 10.3969/j.issn.0529-1356.2010.05.017 Abstract ( )   Objective To explore the expression of trefoil factor 2(TFF2) in submaxillary glands during the self healing of experimental gastric ulcer in rats. Methods Totally 48 male SD rats were randomly divided into gastric ulcer group(EM>n/EM>=42)and normal group(EM>n/EM>=6). The expression of TFF2 peptide and TFF2 gene were detected by the methods of immunohistochemistry and RT-PCR respectively in the gastric ulcer group and normal group. Results TFF2 immunoreactive cells were mainly located in the cytoplasm of the epithelial cells in the ducts,such as intercalated duct,striated ducts, interlobular duct, and which also showed positive expression in the cavity,even near the cavosurface. TFF2 peptide also showed strong expression in few granule cells or in those cells without granules, and which were weakly positive or negative in the cells with many granules.The integral absorbance values(IEM>A/EM>) and area density(AD) values obviously increased on the day 1 after gastric ulcer(EM>P/EM><0.01),and which was the lowest on the day 2 (EM>P/EM>>0.05),then gradually increased again on the day 4 and day 6 (EM>P/EM><0.01) ,and kept on a high level till 10 days to 23 days compared with the normal group(EM>P/EM><005). The IEM>A/EM> of TFF2/GAPDH in 1,4,6,10,14 and 23 days gastric ulcer group were obviously higher than that of the normal group except the 2\|day gastric ulcer group（EM>P/EM><0.01 or EM>P/EM><0.05）. Conclusion TFF2 in rat submaxillary glands may participate in the regulation of self healing in gastric ulcer. Related Articles | Metrics

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Expression of GATA-4 in ovary tissue of rat model of letrozole-induced polycystic ovary syndrome 2010, 41 (5):  728-732.  doi: 10.3969/j.issn.0529-1356.2010.05.018[HT] Abstract ( )   Objective To investigate the role of GATA-4 in the pathogenesis of polycystic ovary syndrome (PCOS), and to provide some experimental data for the application of the rat model induced by letrozole. Methods Forty female rats were randomly divided into two groups, PCOS model group and control group. The rats in model group were administered letrozole with the dosage of 1mg/(kgI>&#/I>8226;d) dissolved in 10g/L carboxymethyl cellulose（CMC） by P.O./Q.D. for 21 days, and rats in the control group were administered with equal dosage and volume of CMC by P.O/Q.D. for 21 days. Gonadal hormone concentrations in serum were determined by radioimmunoassay; the histological changes in ovarian tissues were observed with HE staining, and the expression of GATA-4 in ovary tissue was detected by immunohistochemistry, real\|time PCR and Western blotting. Results With the comparison of two groups, the concentrations of estradiol and progesterone were found considerably reduced in the control group （EM>P/EM><0.05）, while the serum testosterone, follicle\|stimulating hormone(FSH) and luteinizing hormone(LH) levels were found increased markedly in the model group （EM>P/EM><0.05）. The ovaries in the model group showed high incidence of subcapsular ovarian cyst and capsular thickening and decreased number of corpora lutea comparing to those in the control group. The expression of GATA-4 in preantral follicle and antral follicle granulosa cells in the model group was significantly higher than those in the control group.With the observation on and comparison of staining densities, the mRNA transcription and protein depression of GATA-4 were significantly stronger in the model group than those in the control group（EM>P/EM><0.05）. Conclusion The extra higher expression of GATA-4 in follicle granulosa cells may be significantly correlated with the pathophysiology mechanism of PCOS. The Related Articles | Metrics

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P>Dermatoglyphic patterns of hands in breast cancer/P> 2010, 41 (5):  733-736.  doi: 10.3969/j.issn.0529-1356.2010.05.019 Abstract ( )   Objective To investigate the relationship between digital dermaglyphic patterns of hands and breast cancer. Methods Using anthropometry,the frequency of finger print patterns of 224 women from Ningxia Han nationality (breast cancer and control sample ,112 each) were studied. The differences of them were also analyzed. Results Breast cancer patients had higher digital frequencies of whorls in both hands as compared with that of controls, and it showed a significant differences(EM>P/EM><0.001). The digressive order of both groups was Ⅳ> I> II> III> V. The frequency of 6 or more digital whorls in breast cancer was higher than that of the controls, and it also showed significant differences (EM>P/EM><0.01). Conclusion The frequency of digital whorls (W) may be one of the important early diagnosis markers of breast cancer. Related Articles | Metrics

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Crown and cusp areas of upper and lower first molars of recent central Chinese population 2010, 41 (5):  737-744.  doi: 10.3969/j.issn.0529-1356.2010.05.020 Abstract ( )   Objective Total crown basal area (TCBA) and relative basal cusp area (RBCA) of 282 MSUP>1/SUP> and MSUB>1/SUB> from archeological sites spanning from Neolithic time to Qing dynasty (5 000\|200 years BP) were measured and analyzed to obtain tooth size and variations of recent Chinese populations in central China. Methods With the same standard, the high resolution pictures of occlusal surface were taken for each tooth. Digital image technique and morphometric analysis were used to obtain the TCBA and RBCA for each tooth. Results The RBCAs exhibits a sequence of protocone >paracone> metacone > hypocone in MSUP>1 /SUP>and protoconid > metaconid > hypoconid> entaconid >hypoconulid in MSUB>1/SUB>; the RBCA variations of MSUB>1/SUB> are collectively larger than those of MSUP>1/SUP>. From Neolithic time to Qing Dynasty, the MSUP>1/SUP> and MSUB>1/SUB> TCBA reduced about 6.57% and 6.15% respectively. In contrast, the RBCAs of either MSUP>1/SUP> or MSUB>1/SUB> stay relatively stable in the same period of time. Conclusion Change of food structure is thought to be the most probable reason accounting for this change in consideration of the archeological sites background. The RBCA now presented by MSUP>1/SUP> and MSUB>1 /SUB>of modern humans formed at least 5 000 years ago. Related Articles | Metrics

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Regression analysis of male mandible and skull 2010, 41 (5):  745-750.  doi: 10.3969/j.issn.0529-1356.2010.05.021 Abstract ( )   Objective To study and establish the regression equation applied to estimate the main general skull index with the mandible index, in order to recovery the fragmentary skull. Methods Totally 135 adult male mandibles were used for 35 items of measurements, with the physical anthropology measuring method. Suppose the skull index as dependent variables ySUB>1-26/SUB>, and the mandible index as independent variables xSUB>1-9/SUB>, to study the data with frequencies, erosstabs and regression analysis by using statistical software (SPSS 17.0). Results Totally 180 onefactor linear regression equations were established. After the evaluation, it was proved that the equations had statistical value. Therefore, there were 69 partnership equations in condition of EM>r/EM> ≥ 0.40, EM>P /EM>0.001. Conclusion These equations can be used to estimate the main general skull index with mandible index, to provide evidences for the recover Related Articles | Metrics

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Digit ratio of Dalian Han people 2010, 41 (5):  751-755.  doi: 10.3969/j.issn.0529-1356.2010.05.022 Abstract ( )   Objective To investigate the digit ratio and the distribution characteristics of Dalian Han adult people with direct measuring method and to accumulate data in our country. Methods Under the permission of the objects, 430 healthy adult Han nationality people (220 of male and 210 of female) were randomly selected from Dalian Second People Hospital. The anthropometry methods were adopted to measure the length of the 2nd to 5th digit of left and right hand (palmar aspect) and the digit ratio was calculated and compared with the difference of gender and region. Results 1.The digit ratio: the values of 2D∶3D，2D∶4D，2D∶5D，3D∶4D，3D∶5D，4D∶5D of left and right hand of Dalian Han nation people were 0.899 5，0.901 1，0.965 0，0.965 9，1.207 4，1.206 3，1.073 1，1.072 2，1.343 1，1.339 4，1.252 2，1.249 9 in male and 0.906 5，0.907 9，0.976 6，0.975 3，1.223 0，1.222 6，1.078 1，1.075 4，1.352 7，1.350 2，1.255 1，1.255 8 in female. Both male and female presented the trend of 3D∶5D>4D∶5D>2D∶5D>3D∶4D>2D∶4D>2D∶3D. 2. The gender differences: There were significant gender differences of the digit length and digit ratio in the left and right hands (P&l Related Articles | Metrics

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Physique characteristics of the Han nationality adults living in rural area of Liaoning province 2010, 41 (5):  756-760.  doi: 10.3969/j.issn.0529-1356.2010.05.023 Abstract ( )   Objective To investigate the physique characteristics of the adult of the Han nationality living in rural area of Liaoning province. Methods The method and criterion of the investigation referred the to the regulation in the Method of Anthropometry and the Manual of Anthropometry. The study involved 1 086 Han nationality adults (man 521, woman 565) and 17 somatoscopic measurements，69 anthropometric measurements，and 30 physical indices and index types. Results Most people had upper eyelid eye fold, which was observable in 66.16％ and 71.42％ for males and females respectively. Nearly a quarter people had Mongolian pleats. Black hair and yellow complexion were common，and no significant gender difference. The cephalic index fell into the following categories: Special round head, ultra-narrow facetype and narrow nasal type account for a large proportion. General somatotype features of the body included: medium and long trunk type，broad chest circumference，medium breadth of the shoulder and moderately broad breadth of pelvis, moderate brachyskelic and moderate long leg were common.Average male height was 1 667.10mm, and women average height was 1 552.80mm, and different age group varied significantly(EM>P/EM>﹤0.05) with increasing trend. Conclusi Related Articles | Metrics

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Human embryonic stem cells line chHES-20 expresses germ cell differentiation associated genes 2010, 41 (5):  761-763.  doi: 10.3969/j.issn.0529-1356.2010.05.024 Abstract ( )   Objective To investigate whether the human embryonic stem cell line chHES-20 can express the germ cell differentiation associated genes. Methods RT-PCR was used to detect the expression pattern of germ cell differentiation associated genes in chHES-20 cell line. The present sdudy used the normal human testis and human embryonic fibroblast cell line to serve as the positive and negative controls, respectively. Results The pluripotency and self-renewal specific genes OCT4,NANOG and SOX2 were able to be expressed in chHES-20 cell line.The DAZL(deleted in azoospermia-like) and FRAGILIS of pre-meiotic germ cell-specific genes were expressed in chHES-20 cell line. chHES-20 was not able to express the meiosis specific gene SCP3(synapto complex protein 3 gene) and germ cell pos-meiotic differentiation gene VASA. Conclusion chHES-20 embryonic stem cell line possesses the potential to differentiate germ cell. Related Articles | Metrics

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Expression of fibroblast growth factor 10 during the development of mouse ovaries 2010, 41 (5):  764-767.  doi: 10.3969/j.issn.0529-1356.2010.05.025 Abstract ( )   Objective To examine the expression of fibroblast growth factor 10 (FGF10) during the development of mouse ovaries. Methods Ovaries were isolated from mice at gestation day14.5, day16.5, day18.5, newborn and three-week-old. The samples of each group were six. FGF10 protein was detected in mice ovaries by immunohistochemistry.Three-week-old mice were treated with pregnant mare serum gonadotrophin(PMSG), by intraperitoneal injection and forty-eight hours after human chorionic gonadotrophin ( hCG ) was injected into those mice, mRNA was extracted after three hours of injection hCG，the expression of FGF10, luteinizing hormone receptor(LHR), FGF7 and fibroblast growth factor receptor 2Ⅲb(FGFR2Ⅲb) mRNA in ovaries were compared by RT-PCR. Results FGF10 protein expressed in the oocytes in different developmental stages of mouse ovary, and the highest level of staining was observed in the oocytes of the newborn mouse. After gonadotrophins injection, RT-PCR results showed that the expression of FGF10 mRNA decreased, but LHR increased, and FGF7 and FGFR2Ⅲb mRNAs levels unchanged. Conclusion FGF10 is localized on the cell membrane of oocytes of mouse， and gonadotrophins affect the expression of FGF10 mRNA in the ovary of mouse. These results indicate that FGF10 may be connected with regulating o Related Articles | Metrics

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Biocompatibility of optimized acellular dermal matrix by glutaraldehyde crosslinking 2010, 41 (5):  768-773.  doi: 10.3969/j.issn.0529-1356.2010.05.026 Abstract ( )   Objective To select the best glutaraldehyde cross-linkage by implantation experiment and obtaining the optimizing acellular dermal matrix(ADM) by the method of NaOH maceration and the best glutaraldehyde cross-linkage. Fibroblasts(FBs) were cultured on optimized ADM and observed to find a good biocompatibility. Methods We prepared rat ADM by the method of NaOH maceration. The resulting ADM was divided into four groups by different cross-linkage, including group A(0min), group B(10min), group C(15min) and group D(30min). We compared physical character, area after implantation, inflammation reaction and the number of infiltrated FBs and blood vessels of four groups by ordinary and histological observation in order to select the best cross-linkage. The cytotoxicity of ADM was evaluated by methyl thiazolyl tetrazolium (MTT) colorimetry. FBs were cultured on optimized ADM and observed by immunohistochemistry and transimission electron microscope(TME). The expression of type Ⅰand Ⅲ precollagen mRNA in the FBs was examined by RT-PCR. Results Four weeks after implantation, areas of uncross-linked ADM were decreased and the ADM closely adhered to the tissue around, inflammation cells could be observed. There were a few inflammation cells and statistical negative correlation between the number of infiltrated cells/blood vessels and cross-linkage in the crosslinked ADM. The best cross-linkage time was 10 min. The cytotoxicity scores of optimized ADM were grade 0 or 1. FBs grew and proliferated well in the optimized ADM. Compared with the one grown on the plate, the expression of type Ⅰand Ⅲ precollagen mRNA of FBs grown in the optimized ADM were decreased. Conclusion The process of NaOH maceration is simple and low cost. The optimized ADM for 10minutes has good biocompatibility and may provide space and transduct information for cells growth. It effers a molecular biology foundation which may reduce scar formation because of lowly regulating m Related Articles | Metrics

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Application of intravital fluorescent staining with tissue clearing in displaying microvascular architecture of mouse skin 2010, 41 (5):  774-776.  doi: 10.3969/j.issn.0529-1356.2010.05.027 Abstract ( )   Objective To develop whole-mount tissue clearing methods for the cutaneous microvessel in situ visualization. Methods Twenty ICR mice were randomly divided into 2 groups: Ⅰ and Ⅱ. Our approach included labeling luminal surfaces of endothelial cells by i.v injection of fluorescein isothiocyanate(FITC) conjugated Anti-CD31, ulex europaeus agglutinin-I(UEA-I) respectively, and then subdivided them into unclearing group (a) and clearing group (b). The former was observed using a fluorescence microscope after frozen section, while the latter was observed followed by optical clearing of whole-mount tissue to reduce light scatter and permit visualization of microvessel morphology. Results GroupⅠa andⅡa had lower vessel density, the angiosomes were damaged or destroyed obviously；GroupⅠb andⅡb presented with limpid and integrity angiosomes. This optical clearing technique allows imaging of the microvessel by microscopy, to depths of up to 1 000μm below the specimen surface. Conclusion The application of intravital fluorescent staining wit Related Articles | Metrics