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2012, Volume 43 Issue 4
06 August 2012 Previous Issue    Next Issue
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综述
神经生物学
细胞和分子生物学
肿瘤生物学
解剖学
组织学胚胎学发育生物学
人类学
技术方法
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综述

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Primary analysis of current studies on the digit ratio 2012, 43 (4):  0-573.  doi: 10.3969/j.issn.0529-1356. 2012.04.024 Abstract ( )   The digit length ratio of the second finger length to fourth finger length (2D∶4D) is a putative marker for prenatal androgen action, exists sexual, ethnic, racial, and geographic differences,is associated to the incidences of diseases and may pre-estimate physical capacity potential. This study reviewed publications on digit ratio over the past decade and analysed their publication time, professional distribution and impact factors. It indicated that the major publications came from British, United States, Austria and Germany and few from China. In addition in humans, digit ratio was studied in the primates. The digit ratio, as a anthropometry index, still is a hotpoint as it is important to estimate health risk factors, prevent some diseases, study human behavior and cognizence and so on. In the future, it needs to be probed thoroughly in differences of intergroups digit ratio, animas digit ratio, applications in medicine, sport science, behavior science and foundation m Related Articles | Metrics

神经生物学

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Effects of transplantation with bone marrow-derived endothelial progenitor cells on learning, memory and neurons in the cortex of the parietal lobe after cerebral ischemia reperfusion injury of atherosclerotic model rats 2012, 43 (4):  433-438.  doi: 10.3969/j.issn.0529-1356.2012.04.001 Abstract ( )   Objective To study behavior abilities and morphological changes on neurons in the cortex of parietal lobe after cerebral ischemia reperfusion injury (IRI) of atherosclerotic(AS) model rats and observe the effect of transplantation with bone marrow-derived endothelial progenitor cells (EPCs) on the AS model rat. Methods A total of thirty male adult Wister AS model rats were established by fat-rich diet feeding for six consecutive weeks. EPCs were obtained from the bone marrow and the cells cultured in vitro in M199.On the 7th day, middle cerebral artery occlusion (MCAO) rat models were established by the method of thread thrombus. One day after MCAO, the rats were randomly divided into the EPCs transplant group (the EPCs were injected into the caudal vein), the AS group and the IRI group (the same volume of PBS was injected into the caudal vein).The learning and memory abilities were detected by they-maze after seven days. Then, the expression of vascular endothelial growth factor (VEGF) mRNA was detected by reverse transcription polymerase chain reaction and the ELISA method was used to detect the VEGF content. Caspase-3 and GFAP immunopositive cells in the cortex of the parietal lobe were observed under a light microscope, and quantitative analysis was performed by cell morphometric technique. BR>Results EPCs from bone marrow were isolated, induced and cultured successfully in vitro . Following culture for 24 hours, adherent cells presented spindle-shaped appearance. Cell colony-forming units appeared 72 hours after seeding and increased obviously after five days. One week later the cells confluenced to 80%. Attached cells formed a cobblestone-like structure by 10-14 days. Observation using fluorescence microscopy, the double-positive staining with DIL-ac-LDL and FITC-UEA-1 cell population was above 75% among adherent cells. Compared with the IRI group, the learning and memory abilities of the EPCs transplant group were obviously improved, but the content and mRNA of VEGF were significantly decreased (EM>P/EM> <0.05),the quantity of Caspase-3 and GFAP immunopositive neurons were obviously decreased in the EPCs transplant group (EM>P/EM> <0.05). Conclusion EPCs from bone marrow efficiently promote neurological functional recovery and decrease the pathological lesion of the cortex in the parietal lobe after cerebral IRI of AS model rats, which may improve the neovascularization, reduce infarct area and improve neurocrine function. BR> Related Articles | Metrics

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Expression change of the secreted frizzled-related protein 4 in spinal cord of amyotrophic lateral sclerosis transgenic mice 2012, 43 (4):  439-444.  doi: 10.3969/j.issn.0529-1356.2012.04.002 Abstract ( )   Objective To study the expression change of sFRP4 in the spinal cord of adult amyotrophic lateral sclerosis(ALS) transgenic mice, and explore the role of sFRP4 in the pathogenesis of ALS. Methods Both the ALS transgenic mice and wild-type mice were selected, some were perfused intracardially with 4% paraformaldehyde at the age of 95days, 108days and 122days.Their spinal cords were dissected and sectioned. Some mice were anesthetized and spinal cords were quickly removed. RNA and total protein were obtained. The distribution of sFRP4 positive cells, and expression changes of sFRP4 mRNA and protein in the spinal cord of adult ALS transgenic mice and wild-type mice were observed by immunohistochemical techniques, RT-PCR and Western blotting at different time points. Results Compared with wild-type mice, sFRP4 mRNA and protein expression were all increased significantly in the spinal cord of ALS transgenic mice at the age of 95days, 108days, and 122days(EM>P /EM>< 0.05).The majority of sFRP4 positive cells were located in the ventral horn of the gray matter. Nerve fibers were also positive within the white matter. Compared with wild-type mice, sFRP4 positive cells were increased distinctly. sFRP4-labeled cells were co-expressed GFAP and β-tubulinIII in the ALS transgenic mice and wild-type mice, and sFRP4/GFAP double positive cells were increased in ALS transgenic mice.Conclusion BR>sFRP4 positive cells were increased significantly, and the expression of sFRP4 mRNA and protein were increased in the spinal cord of ALS transgenic mice, suggesting that sFRP4 may be closely related to the pathogenesis of amyotrophic latera Related Articles | Metrics

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Protective effect of lycium barbarum polysaccharides on methyl mercury chloride-induced hippocampal neural stem cells injury 2012, 43 (4):  445-450.  doi: 10.3969/j.issn.0529-1356.2012.04.003 Abstract ( )   Objective To determine the protective roles of lycium barbarum polysaccharide on methyl mercury chloride-induced injury for neuronal differentiation of hippocampal neural stem cells(NSCs). Methods NSCs were collected from the hippocampus of 16-embryonic day Sprague-Dawley rats. The cells were cultivated in neural stem cell-specific medium for 10 days. The neurospheres were dissociated into single cells and cultured in the 24-well plates with poly-L-lysine-coated cover glass. Cells were divided into four groups: a control group in which the cells were cultured in DMEM/F12 medium; a lycium barbarum polysaccharide group-the cells were cultured in DMEM/F12 with lycium barbarum polysaccharide; a methyl mercury chloride group-the cells were cultured in DMEM/F12 with methyl mercury chloride; Methyl mercury chloride + lycium barbarum polysaccharide group-the cells were cultured in DMEM/F12 with methyl mercury chloride and lycium barbarum polysaccharide. The cell growth and differentiation were determined by immunostaining against MAP-2 or GFAP antibody.Results The percentage (3.63%±0.62%) and average perimeter (63.36μm±5.57 μm) of the differentiated neurons in the methyl mercury chloride group were lower than that in the control group, while the percentage (7.75%±0.59%) and average perimeter (253.3μm±11.21μm) of them in lycium barbarum polysaccharide group were much more than all other groups. Compared with the methyl mercury chloride group, after methyl mercury chloride treatment the lycium barbarum polysaccharide increased the neuronal differentiation (5.92%±0.98%) to the level in control group and their average perimeter (111.9μm±6.07μm) in this group also showed 2-fold increase. Conclusion Lycium barbarum polysaccharide promotes the NSCs to Related Articles | Metrics

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Sodium 4-phenylbutyrate ameliorates traumatic brain injury in rats by reducing endoplasmic reticulum stress 2012, 43 (4):  451-457.  doi: 10.3969/j.issn.0529-1356.2012.04.004 Abstract ( )   Objective To investigate the expression of the endoplasmic reticulum stress-associated proteins glucose regulated protein 78 (GRP78), phosphorylation pancreatic ER kinase (p-PERK) and C/EBP homologous protein (CHOP) in rats after traumatic brain injury (TBI), and the mechanism of sodium 4-phenylbutyrate (4-PBA) on neuronal injury by reducing endoplasmic reticulum stress after TBI. Methods A total of 114 male Sprague-Dawley rats were randomly divided into 3 groups: the sham, TBI, TBI plus 4-PBA groups. A rat model of diffuse brain injury was established according to Marmarou’s falling model. The rats in 4-PBA group were treated with 4-PBA (120 mg/kg) i.p. immediately after TBI, once a day for 3 days. At 3, 6, 12, 24, 48 and 72 hours after injury, the brain tissue samples were prepared for examination of brain pathological changes, and determination of the expression of GRP78, p-PERK and CHOP, by means of immunohistochemistry and Western blotting. Behavioral tests were performed at 24, 48 and 72 hours after injury. Results Compared with the sham group, GRP78, p-PERK and CHOP were significantly increased in the TBI group. GRP78 began to increase at 3 hours after injury, peaked at the 12 hours, and then gradually returned to the basic level. P-PERK peaked at 12 hours, whereas, CHOP peaked at 24 hours,and then reduced at 48-72 hours,but still was higher than the basic level. At the corresponding times, compared with the TBI group, the expression of GRP78, p-PERK and CHOP decreased remarkably in the 4-PBA group（EM>P/EM> <0.05）. The scores of behavioral test were significantly increased in 4-PBA group,compared with TBI group（EM>P/EM> <0.05）. Conclusion Endoplasmic reticulum stress is started after traumatic brain injury in rats. 4-PBA has a protective effect on TBI, which ma Related Articles | Metrics

细胞和分子生物学

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Comparative analysis of gene expression profiles between liver cirrhosis and liver regeneration in rats 2012, 43 (4):  458-467.  doi: 10.3969/j.issn.0529-1356.2012.04.005 Abstract ( )   Objective To explore the relevance of rat liver cirrhosis (LC) to rat liver regeneration (LR). Methods The model of CCl4-induced LC was established by using 24 adult SD male rats (6 rats in each group). 114 adult SD male rats were randomly divided into 19 groups consisting of 9 partial hepatectomy groups, 9 sham operation groups and 1 normal control group, and 2/3 partial hepatectomy-induced LR was established. Then Rat Genome 230 2.0 array was used to detect gene expression profiles of livers tissues obtained from the above two models. Some methods of bioinformatics and systematic biology were applied to uncover the correlation of gene expression changes and physiological activities between the LC and LR. The reliability of the array data was confirmed by real-time polymerase chain reaction (RT-PCR). Results It was found that 304 genes were changed significantly in expression in rat LC occurrence and 948 genes in rat LR. Of them, 183 genes were the LC-specific, 827 genes were the LR-specific, and 121 genes were shared in both. H-clustering analysis demonstrated that physiological activities of LC had no correlation with those of LR in time. K-means cluster analysis revealed that gene expression trends of Cluster 1-5 (C1-5) groups were similar in LC and in LR, and that those of their C6 group were contrary with the gene expression changes of LR more abundant. Gene ontology(GO) classifications and functional cluster analysis found that physiological activities including immune and inflammatory response, cell migration and adhesion were increased both in LC and in LR, whereas various metabolic activities were decreased. Among them, reaction of liver to stimulation in LC was stronger than in LR in C 2, but in C 6 showed a contrary result. At the same time, DNA repair, cell proliferation, lipid metabolism, homeostasis, oxidation reduction etc in LC were weaker than in LR. BR>Conclusion The changes in gene expressions and physiological activities of LC and those of LR have not only similarities but also differences. BR> Related Articles | Metrics

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Quality assessment of report gene green fluorescence protein in chicken embryo EM>in vivo/EM> electroporation 2012, 43 (4):  468-472.  doi: 10.3969/j.issn.0529-1356.2012.04.006 Abstract ( )   P>Objective The method of in vivo electroporation has been set up successfully and we further analyzed the effect of report gene green fluorescence protein(GFP) on the morphology of developing chicken embryos after in vivo electroporation, and also analyzed the expression of α-smooth muscle actin (α-SMA) and neurofilament during chicken embryonic development. Methods pCAGGS-GFP was transformed into chicken embryos with in ovo culture at 3days and ex ovo culture at 3-5days. In 24hours after in vivo electroporation, GFP-positive embryos were selected under stereo fluorescence microscope, and the GFP-negative embryos served as controls. Five embryos were analayzed for each group. Fluorescence immunohistochemistry was applied to analyze the expression ofα-SMA and neurofilament in chicken spinal cord and tectum. Results At different stages of chicken embryos and different time after in vivo electroporation, the expression of α-smooth muscle actin and neurofilament did not show difference in experimental group and wild type, as well as in GFP-positive area and GFP-negative area. The morphology of embryos was not changed after electroporation with pCAGGS-GFP either. BR>Conclusion GFP as a report gene to in vivo electroporation for chicken embryos does not affect the expression of α-smooth muscle actin and neurofilament, as well as no effect on the morphology of chicken embryos, so GFP Related Articles | Metrics

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Replicative senescence characteristics of human leukemia cell line (K562) induced by ginsenoside Rg1 2012, 43 (4):  473-478.  doi: 10.3969/j.issn.0529-1356.2012.04.007 Abstract ( )   Objective To discuss the characteristic changes of leukemia cell line K562 on replicative senescence induced by ginsenoside Rg1. Methods The effect of Rg1 on the leukemia K562 cell line proliferation was detected by MTT colorimetric test in order to screen optimal time and drug concentration induced cells senescence. The flow cytometry method was used to analyze the cell’s cycle. The percentage of positive cells, the telomere length and the expression of the senescence -related proteins P21,P53,Rb were detected by SA-β-gal staining, southern blotting and western blotting methods, respectively. The ultrastructural senescence changes were observed under the a transmission electronic microscope. Results The optimal time and concentration in order to inhibit the proliferation of K562 cells were 48hours and 20mol/L respectively. The K562 cells arrested G2/M phase. The percentage of positive cells was increased (EM>P/EM> <0.05 ). The senescence -related proteins were up-regulated (EM>P/EM> <0.05). The telomere length became shorten quickly ( EM>P/EM> <0.05 ). Conclusion Rg1 may induce leukemia cell line K562 into the state of replicative senescence by the ce Related Articles | Metrics

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Direct cloning of immunoglobulin kappa light chain variable genes from rabbit peripheral blood 2012, 43 (4):  479-483.  doi: 10.3969/j.issn.0529-1356.2012.04.008 Abstract ( )   Objective To amplify immunoglobulin Kappa light chain variable genes rapidly from rabbit peripheral blood. Methods PCR primers were designed according to the cDNA sequence of rabbit germline IGKV, IGKJ and IGKC genes, which were obtained from IMGT/GENE-DB. With the total RNA extracted from rabbit peripheral blood as template, an one-step RT-PCR reaction was done, and the products were cloned to T vector, sequenced, compared with rabbit germline Kappa light chain variable genes in IMGT/V-QUEST. By counting the kinds of Vκ-Jκ-Cκ assembly, the specificity and compatibility of the primers were evaluated. Results Of 4 pairs of primers designed, RK.C1［1-2-7］, RK.C1［3-6-8］and RK.C2［1-2-3-4］, but not RK.C1［4-5-9］were succeeded in amplifying the rabbit total RNA. Fifty-one clones from their products all belonged to rabbit immunoglobulin Kappa light chain variable and constant genes and different each other. Among them, 33 clones’constant regions were encoded by IGKC1*1, the rest, 18 clones, were encoded by IGKC2*03, and up to 22 of 68 IGKV and only 1 of 8 IGKJ functional germline genes appeared. Totally, 17 kinds of Vκ-Jκ-IGKC1 and 11 kinds of Vκ-Jκ-IGKC2 assembly were found in these 51 clones. IGKV1S10*01 was found in 12 clones, including 7 Vκ-Jκ-IGKC1 and 5 Vκ-Jκ-IGKC2 clones, which was very higher than other Vκ genes； There were existed some differences in sequence, although the 12 clones were from the same Vκ germline ge Related Articles | Metrics

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Differentiation of rat bone marrow mesenchymal stem cells into cardiomyocytes induced by exogenous Nkx2.5 gene 2012, 43 (4):  484-489.  doi: 10.3969/j.issn.0529-1356.2012.04.009 Abstract ( )   Objective To investigate the feasibility of transfecting bone marrow mesenchymal stem cells (BMSCs) with Nkx2.5 to enhance cardiomyogenic differentiation. Methods Rat BMSCs were isolated by the whole bone marrow culture and amplified by serial subcultivation. CD90 and CD45 of the third passage cells were identified by flow cytometry. Using the cationic liposome reagent, Lipofectamine 2000, the plasmid pEGFP-N1-Nkx2.5 was transfected into BMSCs. The transfected cells were observed under an inverted fluorescence microscope and expression of Nkx2.5 was examined with immunocytochemistry. Expression of cardiac troponin T(cTnT) and GATA4 were determined with Western blotting and immunocytochemistry. Results 99% of the third passage cells were positive for CD90 and negative for CD45 on flow cytometry. After 48 hours of transfection, some cells in the transfected group expressed the green Nkx2.5-EGFP fusion protein and Nkx2.5 was expressed only in pEGFP-N1-Nkx2.5 transfected group (n =3). After 4 weeks, Western blotting results indicated that the expression of cTnT in pEGFP-N1-Nkx2.5 transfected group was significantly higher than those in pEGFP-N1 vector transfected group and in control one (EM>n/EM> =3). Immunocytochemistry results showed that the expressions of cTnT and GATA4 in pEGFP-N1-Nkx2.5 transfected group were respectively the highest in the three groups ( EM>n/EM> =3). Conclusion Exogenous expression of Nkx2.5 gene may enhance the cardiomyogenic differentiation of BMSCs. Related Articles | Metrics

肿瘤生物学

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Changes in expression and localization of heat shock protein 70 during curcumin-induced apoptisis of human esophageal cancer cell line EC9706 2012, 43 (4):  490-499.  doi: 10.3969/j.issn.0529-1356.2012.04.010 Abstract ( )   P>Objective To investigate the effect of curcumin on human esophageal cancer EC9706 cells and explore the role of heat shock protein(HSP70) in cell apoptosis by examining changes in the nuclear matrix and its relationship with apoptosis-related proteins. Methods Cell counting and flow cytometry were performed to probe the inhibitory effect of curcumin on cellular proliferation. Transmission electron microscopy and optical microscopy were used to observe the structural changes in EC9706 cells before and after apoptosis. Agarose gel electrophoresis was conducted to investigate the DNA structure of EC9706 cells before and after apoptosis. Two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and mass spectrometry (MS) analysis were performed to investigate the presence and changes of HSP70 in the nuclear matrix of EC9706 cells before and after curcumin treatment, which was further corroborated by Western blotting assay. Laser confocal scanning microscopy was used to observe the colocalization of HSP70 with Bax and Bcl-2 during apoptosis. Results BR>The results indicated that curcumin could markedly inhibited EC9706 cell proliferation and finally induced apoptosis. Data from 2-D PAGE, MS, and Western blotting showed that HSP70 was involved in the nuclear matrix proteins and expression of HSP70 was downregulated after curcumin treatment. Laser confocal microscopy showed that HSP70 colocalized with Bax and Bcl-2, and the colocalized regions were altered by the curcumin treatment. Conclusion Our work proves that curcumin could definitely induce EC9706 cells into apoptosis. As a new found nuclear matrix protein, the expression and distribution of HSP70 are altered during the apoptosis of EC9706 cells. The colocalization of HSP70 with apoptosis-related genes evidently affects the apoptosis of EC9706 cells. BR>/P> Related Articles | Metrics

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Tumor stem cell spheres generated from human lung cancer cells NCI-H446 2012, 43 (4):  500-505.  doi: 10.3969/j.issn.0529-1356.2012.04.011 Abstract ( )   Objective To isolate and expand cancer stem cells in human lung cancer cell line NCI-H446 and to identify their biological properties. Methods NCI-H446 cells were cultured in serum-free conditions to generate tumor spheres. The tumor spheres were then expanded and cultured in a serum-containing medium to permit their differentiation.The proliferative capacity of the normal NCI-H446 cells and tumor sphere-forming cells were tested by MTT assay, and the tumorigenicity and invasion of the both normal and sphere-cells were evaluated using animal and Transwell invasion experiments.The expression of CD133 and CD44 on the surface of the normal NCI-H446 cells and tumor sphere-forming cells were detected by flow cytometry. Results A small number of floating tumor spheres were isolated and expanded in serum-free conditions and the cell of the spheres maintained a strong capacity for self-renewal and proliferation. These tumor spheres attached to the bottom of culture plates and began to differentiate in a serum-containing medium.The xenograft tumorigenicity of tumor sphere forming cells was significantly higher than that of the normal NCI-H446 cells while injecting 5×105 cells in animal experiments.The capability of invasion of tumor sphere forming cells was significantly higher than that of the normal NCI-H446 cells in transwell invasion experiments. The percentage of CD133,CD44 cell population was significantly higher in tumor spheres than that in the normal NCI-H446 cells. Conclusion Cancer stem cells exist in human lung cancer cell line NCI-H446 and can be isolated and expanded in serum Related Articles | Metrics

解剖学

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Arch morphology of non-syndromic complete unilateral cleft lip and palate patients 2012, 43 (4):  506-509.  doi: 10.3969/j.issn.0529-1356.2012.04.012 Abstract ( )   Objective To study morphology of the maxilary arch of the patients with non-syndromic complete unilateral cleft lip and palate in order to help assessment of clinical development of arch, occlusion and occlusal guidance therapy. Methods BR>This study included maxillary models of 32 non-syndromic complete unilateral cleft lip and palate patients. The models were scanned and measured by Denmark 3Shape Dental Model scanner and software. Length, width and circumference of both noncleft and cleft sides were measured. Spss14.0 was used to analyze the results with paired samples statistics. Results In the premolar area, there was statistical significance between width of noncleft and cleft side (first premolar areaEM> t/EM> =5.19, EM>P/EM> <0.01；second premolar area EM>t /EM>=3.24, EM>P /EM><0.05). In the first permanent molar area, there was no statistical significance between width of noncleft and cleft sides (EM> t/EM> =0.48, EM>P/EM> >0.05). The circumferences of noncleft side was longer than cleft side and the difference was 4.2 mm, which was statistically significant.( EM> t/EM> =4.611, EM>P /EM><0.01). There was statistical significance between them. Conclusion In non-syndromic complete unilateral cleft lip and palate patients, the cleft sides of maxillary arch are significantly under-developed. The circumference of cleft sides is less than n Related Articles | Metrics

组织学胚胎学发育生物学

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Prenatal alcohol exposure induces the mossy cell apoptosis in hippocampus of sphingomyelin synthase 2 knockout mice 2012, 43 (4):  510-517.  doi: 10.3969/j.issn.0529-1356.2012.04.013 Abstract ( )   Objective To investigate the role of ceramide regulation on the alcohol-induced neuronal cells apoptosis in mice. Methods Using sphingomyelin synthase 2 knockout (SMS-/-2) mice and wide-type (WT) mice to establish the model of prenatal alcohol exposure. A total of 360 pups from control the group and ethyl alcohol (EtoH) treatment group was used in the study. The levels of serum sphingomyelin (SM) was detected with enzymatic method in P0. Apoptosis of mossy cells in hippocampus were investigated after alcohol exposure with immunofluorescent labeling. The expression of activated Caspase-8 and activated Caspase-3 in hippocampus were tested with Western blotting analysis. Results In WT and SMS-/-2mice, prenatal alcohol exposure down-regulated the SM levels with dose-dependency ( EM>F /EM>=41.08,EM> P/EM> <0.05). The SM level of serum in SMS-/-2 pups was significantly lower than that of WT pups( EM>F/EM> =53.34, EM>P/EM> <0.01). In both WT and SMS-/-2pups, the number of apoptosis moss cells in hippocampus was increased after prenatal alcohol exposure with dose dependency ( F =15.61, P <0.05), and then decreased with the growing age. However, compared with the WT pups, the number of apoptosis neurons in hippocampus of SMS-/-2 pups increased more than WT mice ( EM>F /EM>=11.72, EM>P /EM><0.05). Western blotting supported the results of immunofluorescent labeling. Conclusion The reduction of SM level in SMS-/-2 mice leads to the ceramide accumulation in brain tissue. Ceramide is involved in prenatal alcohol exposed neuronal apoptosis in the process, and may promote apoptosis. Alcohol exposure during pregnancy, mainly through the death receptor pathway, induces neuronal apoptosis. Related Articles | Metrics

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Prenatal alcohol exposure affects development of retinal bipolar and horizontal cells in mice 2012, 43 (4):  518-523.  doi: 10.3969/j.issn.0529-1356.2012.04.014 Abstract ( )   Objective To investigate effects of alcohol exposure during pregnancy on retinal bipolar and horizontal cells in filial mice.Methods The animal model of prenatal alcohol exposure was made. The development of bipolar cells and horizontal cells at prenatal days 7, 14 and 30 was investigated with immunofluorescent labeling and DiI scattering techniques. Results In all age groups, the density of the bipolar cells was significantly lower in the experiment groups than in the control groups. The dendritic field and the number of branches of horizontal cells were significantly lower in the experimental group than in the control group ( P <0.01). Conclusion Alcohol exposure during pregnancy affects development of retinal bipolar and horizontal cells in mice. Related Articles | Metrics

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Effects of Heshouwuyin on the expression of P450 side-chain cleavage enzyme and steroidogenic acute regulatory protein in testis tissue of the exercised-induced fatigue rat 2012, 43 (4):  524-529.  doi: 10.3969/j.issn.0529-1356. 2012.04.015 Abstract ( )   Objective To explore the effect of Heshouwuyin on the expression of P450 side-chain cleavage enzyme(P450scc) and steroidogenic acute regulatory protein (StAR) in testis tissue of rats with exercised-induced fatigue. Methods Sixty SD rats were randomly divided into 6 groups,10 rats in each group, including a normal control group(group A), a Heshouwuyin administered normal group(group B),a model control group(group C), a spontaneously recovering group(group D), a Heshouwuyin treated group (group E), and a Heshouwuyin prevented group (group F). Groups C, D, E and F were exercised-induced fatigue animal model, groups in which animals were administrated Heshouwuyin ［20g/(kg.d), contained crude drug 4.8kg/L］. Animals in group F were administrated Heshouwuyin before modeling, and animals in group E were treated with Heshouwuyin for 60 days after success of modeling. Beckmancoulter Unicel Dxl 800 was used to detect the level of serum testosterone, according to the manufacture’s instructions. Western blotting and RT-PCR were used to observed the differential expression of P450scc and StAR. Result P450scc protein was expressed in Leydig cells and spermatocyte. Compared with group A, the expression of P450scc protein and mRNA in group C decreased ( P <0.05), and the expression of P450scc protein and mRNA in groups B and F increased more than E, D, A. StAR protein was expressed in Leydig cells. Compared with group A, the expression of StAR protein and mRNA in group C decreased ( EM>P/EM> <0.05), and the expression of StAR protein and mRNA in groups E and F increased more than A, D, but there was no difference between groups A and D ( EM>P/EM> ＞0.05). Conclusion Th Related Articles | Metrics

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Effect of tautomerase activity inhibitor ISO-1 on implantation of the mouse embryo 2012, 43 (4):  530-534.  doi: 10.3969/j.issn.0529-1356. 2012.04.016 Abstract ( )   Objective To investigate the effects of ISO-1,a selective macrophage migration inhibitory factor (MIF) tautomerase activity inhibitor,on the blastocysts implantation in mice and its mechanism. Methods Totally 150 pregnant mice were randomly divided into five groups. Low- dose, middle-dose and high-dose groups were given ISO-1 (2 mg/kg, 6 mg/kg, 18 mg/kg, respectively) by i.p. injection on day 3 of pregnancy and the two control groups were treated with the equal volume of 1% dimethyl sulfoxide (DMSO) or saline. A half of the mice were killed on the day 4 of pregnancy, and the uteri were excised. Structures of the endometrium were observed with HE staining. The expressions of MIF protein and mRNA were studied by the immunohistochemical staining and in situ hybridization techniques,respectively. The remaining half of the mice were killed on day 8 of pregnancy to examine the number of the embryos and calculate the uterine organ coefficient. Results Compared with the controls, there were no significant differences in the number of the embryos and the uterine organ coefficient in the low-dose group and middle-dose group( EM>P /EM>>0.05). In the high-dose group, the number of the embryos increased significantly ( EM>P/EM> <0.05), but there were no significant differences in the uterine organ coefficient compared with the controls( EM>P/EM> >0.05).The histological structure of endometrium showed no significant modifications among the groups. MIF protein and mRNA were mostly expressed in the cell aggregates scattered throughout the stroma. There were no significant differences in the average absorbance of MIF protein and mRNA in all the experimental groups compared with the controls ( EM>P /EM>>0.05). Conclusion ISO-1 may promote mice blastocysts implantation in a dose-dependent manner. ISO-1 may improve blastocysts implantation in mice by antagonizing the biological function of MIF. Related Articles | Metrics

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Expression and significance of trefoil factor 1 in the submandibular gland during the self-healing of experimental gastric ulcer in rats 2012, 43 (4):  535-539.  doi: 10.3969/j.issn.0529-1356. 2012.04.017 Abstract ( )   Objective To explore the expression of trefoil factor1 (TFF1) gene in submandibular glands during the self-healing of experimental gastric ulcer in rats. BR>Methods Immunohistochemical SP and RT-PCR methods were used to detect the expression of TFF1 peptide and TFF1 mRNA in submandibular glands on 42 gastric ulcer(GU),21 normal saline control(NS) and 6 normal(N) rats. Results TFF1 immunoreactive profiles were mainly located in cytoplasma of granular convoluted tubule (GCT) epithelia cells, especially in many granules cell.Substantial amounts of TFF1 were also found in lumen or luminal membrane of striated duct(SD) and interlobular duct(ILD).In the GUs，especially at the 6-day GU, TFF1 in the lumens of SD and ILD increased.The integral absorbance (IA) values of TFF1 increased obviously in gastric ulcer which was stronger than in the other two groups（EM> P/EM> <0.05 or EM>P/EM> <0.01）.On day 1,2,4,6 the IA values of TFF1 peptide increased gradually and kept at the highest level on day 6,then a trend of decrease on day 10-23,but still obviously higher than the control group. The optical density of TFF1/GAPDH mRNA on the gastric ulcer group was obviously higher than that of the saline and control group during day 2 to day 23 after gastric ulcer operation（ EM>P/EM> <0.05 or EM>P/EM> <0.01）。Conclusion The TFF1 plays a promotive function during the self healing of exper Related Articles | Metrics

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Changes of the subchondral plate in the early stage of the experimental osteoarthritis in rabbits 2012, 43 (4):  540-544.  doi: 10.3969/j.issn.0529-1356. 2012.04.018 Abstract ( )   Objective To observe the relationship between the subchondral plate and cartilage in the osteoarthritic rabbit knee, and to investigate the effect of calcitonin on subchondral plate to protect cartilage. Methods Male rabbits were randomly divided into three groups(8 animals per group)，control, model, and experiment groups. In the model and experiment groups，anterior cruciate ligament transaction (ACLT) operation was carried out in the right knee joint. After that, rabbits in the experiment group received subcutaneous injections of calcitonin with 5 IU/(kgI>&#/I>8226;d) dosage. Rabbits in the model group were injected saline. All the rabbits were sacrificed after ACLT operation for 8 weeks. The paraffin sections were stained with HE and von Gieson and graded by Mankin’s scale. Immunohistochemical expressions of matrix metalloproteinases (MMP-13) in cartilage were detected by integral absorbance (IA) analyses. The histomorphometry was applied to analyse the subchondral plate at the proximal tibia. Results Analyzed by the Mankin`s scale in cartilage, the scores of the model group were significantly higher than that of the control and experiment group( P <0.05) and the scores of the experiment group were markedly higher than that of the control group( P <0.05). MMP-13 immunochemical staining in the model group showed a significant overexpression in comparison with those in the control group and experiment group( P <0.05). There was a strong difference of IA scores between the control group and experiment group ( P <0.05); There were significant differences in volume, thickness and porosity of the subchondral plate among the three groups( P <0.05). Conclusion The structures of the subchondral plate and cartilage were destroyed in osteoarthritic rabbit knee induced by ACLT operation, and there was a strong relationship between the subchondral plate and cartilage. Calcitonin treatment on the subchondral plate in the early stage of osteoarthritis may partialy provide protection on Related Articles | Metrics

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Changes of the ultrastructure of colon mucosa after partial hepatectomy in rats 2012, 43 (4):  545-548.  doi: 10.3969/j.issn.0529-1356. 2012.04.019 Abstract ( )   Objective To study the effects of partial hepatectomy (PH) on the ultrastructure of colon mucosa in rats. Methods Thirty-five healthy SD rats were divided into one normal control group, three sham-operation (SO) control groups and three operation (2/3 PH) groups. The rats in the SO and PH groups were sacrificed at the 6th, 12th and 24th hour after speration). The colon was collected and prepared as ultrathin sections. Samples were observed by transmission electron microscope (TEM). Results Compared with that in the normal control group rats, the ultrastructure of colon mucosa did not show distinct changes in the SO groups. However, significant changes were observed in PH 6- and 12- hour groups rats, for example, the microvilli became short, enterocyte interspace was greatly increased, mitochondria of goblet cells was impaired, endocrine cells were stimulated, degranulation of mast cells was marked, macrophages were filled with phagosomes, neutrophilic granulocytes were gathered to lamina propria. In PH 24-hour groups, endocrine cells were still active, but other changes were not evident. Conclusion After PH, colon mucosa barrier is acutely damaged which leads a rising rate of infection. However, stimulation of endocrine cells and mast cells may be helpful to liver regeneration and recovery. Related Articles | Metrics

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Ultrastructural changs of cell necroptosis during renal development in mice 2012, 43 (4):  549-552.  doi: 10.3969/j.issn.0529-1356. 2012.04.020 Abstract ( )   Objective To observe the ultrastructural chang of necroptosis during renal development in mice. Methods Renal necroptosis of three mice aged at embryonic days (E)12,14,16,18 was examined by using electron microscopy. Results There was necroptosis, or programmed necrosis, in the developmental embryonic kidney tissue. EM observations demonstrated that cell necroptosis was similar to cytoplasm appearances of cell necrosis and nuclei of cell apoptosis. Those appearances were swelling cytoplasm with disrupted plasma membrane, and nuclei with shrinkage and margination of chromatin. Conclusion Programmed cell death is composed of necroptosis, oncosis and apoptosis during renal development and apoptosis is more principal. Related Articles | Metrics

人类学

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Correlation between seven length indicators of the head with height in He’nan Han nationality adults 2012, 43 (4):  553-558.  doi: 10.3969/j.issn.0529-1356. 2012.04.021 Abstract ( )   P>Objective To investigate the heads’ morphological characteristics of the Han nationality in He’nan and to analyze the human head length indicators related to their body height. Methods According to the somatometric measurement handbook(2nd Edition), 7 index including head length，head breadth, face breadth，minimum frontal breadth，bigonial breadth，external biocular breadth，and interocular breadth on head of 1 050 men and women in He’nan Han population adults was measured, while the cluster analysis compariton with other domestic ethnic population was implemented, the equation of linear regression was set up. Results Seven head length indicators on Henan Han male adults were higher than the female. There was a significant gender difference in addition to head width and interocular breadth. In the He’nan Han nationality adults, the 7 head length indicators related to the height presented a linear dependence, especially the bigonial breadth. The cluster analysis showed up adoption societies between the He’nan Han nationality adults and Dongxiang nationality, earthy group, the Baoan (Paoan) nationality,and the Miao minority. Conclusion The 7 head length indicators on He’nan Han nationality adults have the characteristics of the North. The height speculatation error is greater according a single length index, yet it is lower us Related Articles | Metrics

技术方法

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Preparation of heart decellularized matrix in rats by perfusing method 2012, 43 (4):  559-563.  doi: 10.3969/j.issn.0529-1356.2012.04.022 Abstract ( )   Objective The aim is to explore an effective method of preparing the whole heart decellularized bio-derived scaffolds (HDBS) in rats for the heart tissue engineering. Methods The hearts were harvested from 30 adult SD rats. HDBS were prepared by the treatment with freeze thawing, typsin, SDS and Triton X-100, and obeserved the decellularization process at the same time. After decellularization, the scaffolds were examined through genomic DNA content analysis, HE staining, scanning electron microscope, transmission electron microscope and immunofluorescence. Results HDBSs maintained the three-dimensional structure with transparent appearance. Based on the DNA quantitative analysis, DNA content of the HDBSs was less than 3% compared to that of the control hearts. HE staining, scanning electron microscope and transmission electron microscope revealed that HDBS were decellularized completely and preserved the full extracellular matrix. Immunofluorescence results showed that collagen, elastin and other extracellular components were preserved completely without obvious nuclear components. Conclusion The HDBS prepared by the treatment with freeze thawing, typsin, SDS and Triton X-100 are decellularized completely, with the full extracellular matrix preserved, which are the ideal three-dimensional bio-derived scaffold material of the heart. Related Articles | Metrics

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Establishment of chicken embryo culture system and EM>in vivo/EM> electroporation methods 2012, 43 (4):  564-568.  doi: 10.3969/j.issn.0529-1356. 2012.04.023 Abstract ( )   Objective To establish the methods of EM>in ovo/EM> and EM>ex ovo/EM> culture of chicken embryo as well as EM>in vivo/EM> electroporation. Methods Fertile eggs were incubated for two to three days（E2-E3）, and in ovo electroporation in spinal cord at E2-E3, and ex ovo electroporation in the tectum of brain at E5-E6 for spatial and temporal gene transformation with the parameter such as volt 18V, current 60mA, pause 90ms, and pulse 60ms for six times were carried out. Results The number of samples were 20 eggs for EM>in ovo/EM> culture and 10 eggs for EM>ex ovo/EM> culture. The survival rate of the embryos at E2-E3 was 85% for EM>in ovo/EM> culture and 80% for EM>ex ovo/EM> culture. The number of samples were 11 in spinal cord at E2-E3 ( in ovo electroporation) and 10 in brain tectum at E5-E6 ( ex ovo electroporation). The positive electroporation rate was 54.5% in spinal cord at E2-E3 (EM>in ovo/EM> ) and 60% in brain tectum at E5-E6 (EM>ex ovo/EM>). Conclusion The methods of in ovo and ex ovo culture of chicken embryos and in Related Articles | Metrics

综述

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Review of the anti-tumor role of onconase 2012, 43 (4):  574-578.  doi: 10.3969/j.issn.0529-1356. 2012.04.025 Abstract ( )   Onconase, also called P30 protein, is a member of RNase A superfamily, and is the only enzyme of this class that reached clinical trials. It has weak RNase activity and strong cytotoxicity to various tumor cells in vitro as well as EM>in vivo/EM>, and it is one of the 100 new drugs in research recently. Onconase was first found in early embryos of leopard frog (Rana pipiens), and onconase can trigger apoptosis of tumor cells via degrading RNA, leading to inhibiting sythesis of protein. In addition, onconase has other anticancer mechanisms, such as influencing expression of specific genes and destroying telomerase RNA. Onconase could also enhance the cytotoxicity of other anticancer drugs. The phase I and phase I/II clinical trials of onconase as a single therapeutic agent in patients with non-small cell lung cancer and other solid tumors is conducting, and the phase III human clinical trials for the treatment of unresectable malignant mesothelioma (MM) is undergoing. Onconase may serve as a second-line therapy for MM. Related Articles | Metrics