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2013, Volume 44 Issue 6
06 December 2013 Previous Issue    Next Issue

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Effects of Lhx8 in hippocampal cholinergic neural regeneration SHI Jin-hong JIN Guo-hua* LI Hao-ming YI Xin QIN Jian-bing TIAN Mei-ling ZHANG Xin-hua 2013, 44 (6 ):  729-734.  doi: 10.3969/j.issn.0529-1356.2013.06.001 Abstract ( )   Objective To investigate the effect of Lhx8 in hippocampal cholinergic neural regeneration of SD rat after fimbria-fornix transection.  Methods According to the different treatments, 72 SD rats were randomly divided into four groups: the normal group, the transected group, the overexpressed group and the transected combined with overexpressed group. On the 28th day after treatments, the sections of rat hippocampus of each group were prepared and stained with immunohistochemical methods to detect the number of newborn Lhx8 and ChAT positive cells. PCR and Western blotting were performed after total RNA and protein of hippocampus were obtained respectively. Results Compared with the normal group, the levels of Lhx8 mRNA and protein were significantly increased in other three groups and the differences were significant among the groups. No significant difference of the levels of ChAT mRNA was observed between the normal group and the transected group while it was significantly upregulated in the overexpressed and the transected combined with overexpressed groups. ChAT protein was detected in the transected, overexpressed and the transected combined with overexpressed groups and the differences were significant among the groups. There were newborn Lhx8 positive cholinergic neurons in hilus and subgranular zone of hippocampal dentate gyrus in the transected, overexpressed and the transected combined with overexpressed groups, with the most in the transected combined with overexpressed group, and the differences were significant among the groups. Conclusion Upregulation of Lhx8 can promote differentiation of neural stem cells into cholinergic neurons in hippocampus significantly, and Lhx8 may be related to cholinergic neural regeneration of hippocampus. References | Related Articles | Metrics

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Lhx8 promotes the differentiation of hippocampal neural stem cells into cholinergic neurons in vitro TIAN Mei-ling ZHU Pei-pei JIN Guo-hua* LI Hao-ming QIN Jian-bing SHI Jin-hong 2013, 44 (6 ):  735-739.  doi: 10.3969/j.issn.0529-1356.2013.06.002 Abstract ( )   Objective To construct eukaryotic expression vector containing full length cDNA of rat Lhx8 gene. Methods  Full length cDNA of rat Lhx8 gene was acquired by PCR method and digested by EcoRI and BamHI. The digested gene fragment was inserted into eukaryotic expression vector pEGFP-C3 and identified by DNA sequencing. The recombinant plasmid was transfected into hippocampal neural stem cells (NSCs) by liposome. Results DNA sequencing demonstrated that the insertion fragment in recombinant plasmid was right, and recombinant plasmid pEGFP-C3-Lhx8 was transfected into NSCs. After 7 days, the number of ChAT positive cells in plasmid transefection group was significantly higher than the negative control group. Conclusion Eukaryotic expression vector pEGFP-C3- Lhx8 is constructed successfully, and Lhx8 promotes the differentiation of NSCs into ChAT positive cellsin vitro. References | Related Articles | Metrics

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Interaction between cell migration and vasculature in the developing cerebellum of the mouse CHEN Wen-jing ZHANG Wen-ling LI Xue WANG Qiang LIU Bin DENG Jin-bo* 2013, 44 (6 ):  740-747.  doi: 10.3969/j.issn.0529-1356.2013.06.003 Abstract ( )   Objective Our aim was to explore the interaction between angiogenesis and neural migration in the developing cerebellum of mice. Methods The immunofluorescence, BrdU and ink perfusion were used to label the radial glial cells, Purkinje cells, granule cells and vasculature in the mouse cerebellum aged from E10 (embryonic day 10) to P90 (postnatal day 90). A total of 146 mice at various ages were used in the study. Results Vascular network began to appear in the cerebellum at about E15. Tith age increasing, vascular density increased. In the meantime, there was close relationship between the radial glial cell development and vascular development. For instance, blood vessels and the projections of radial glial cells in the external granular layer  were arranged regularly and paralleled each other, but the distributions of vessels and radial glial fibers in the internal granular layer and white matter were in disorder, suggesting the interaction between the vasculature and radial glial cells. Finally, the study also showed that many BrdU-positive cells migrated along blood vessels. Conclusion Cell migration plays a crucial role in cerebellar lamellae formation process. We found that the blood vessels not only have interaction with the radial glial cells, but also guide the migration of neurons in the cerebellum, probably serving as scaffolds for the neuronal migration. References | Related Articles | Metrics

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Chronic alcohol exposure induces insulin resistance in mice：the possible mechanism of ceramide ZHAO Jing-ya LU Guang-xiu XU Gao-lei DENG Jin-bo* 2013, 44 (6 ):  748-755.  doi: 10.3969/j.issn.0529-1356.2013.06.004 Abstract ( )   Objective To investigate the relationship among chronic alcohol exposure, insulin sensitivity and ceramide during the alcohol toxicological effect. Methods Wide type (WT) and sphingomyelin synthetase 2 knockout (SMS2-/-) mice (about 3 month old) were used to establish the chronic alcohol exposure model. A total of 90 mice of the two genotypes were divided into the control group, moderate EtOH group and high EtOH group. After alcohol exposure for 5 months, fasting plasma glucose（FPG） and levels of fasting insulin (Fins) were measured. The insulin resistance index (HOMA-IR) was calculated. The expression of insulin receptor substrate 2 (IRS2) in hippocampus was tested with immunofluorescent labeling and Western blotting analysis. Results 1. Alcohol exposure caused WT mice FPG and HOMA-IR index increased with dose-dependency (P<0.05). Comparing with the control group, Fins values increased in the treatment dose group (P<0.05). 2. FPG in SMS2-/- mice increased with dosedependency (P<0.05), but HOMA-IR index had little changes, comparing with WT mice. In addition, the immunohistochemical staining showed that, in both WT and SMS2-/- mice, the number of IRS2-positive cells reduced in CA1 area after alcohol exposure with dose-dependency (P<0.05). However, comparing with the WT mice, the number of IRS2-positive cells of hippocampus in SMS2-/- mice reduced greatly (P<0.05). 3. Immunoblotting evidence of IRS2 supported the results of immunohistochemistry in both WT and SMS-/-2 mice. Conclusion Chronic alcohol exposure can cause insulin resistance in both WT and SMS2-/- mice. IRS2 protein expression decreased with dose-dependency after alcohol exposure. IRS2 loss probably is one of the causes of insulin resistance. Ceramide may be involved in the reduction of IRS2 expression and may promote the formation of insulin resistance after alcohol exposure. References | Related Articles | Metrics

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Temporal and spatial expression pattern of apoptosis and its associated protein in neurons after spinal cord injury in rats DIAO Yao* LIU Xin-ning GE Rui JIAO Ying ZHU Yue 2013, 44 (6 ):  756-760.  doi: 10.3969/j.issn.0529-1356.2013.06.005 Abstract ( )   Objective The purpose of this study is to evaluate temporal and spatial expression pattern of neuron apoptosis and its associated protein in neurons after spinal cord injury in rats. Methods Fifty SD rats were divided into a shamoperated and a spinal cord injury group, 25 rats per group. SCI model was made by clamps methods. The expression profiles were evaluated by counting the cresyl violet-stained surviving cells and the number of TUNEL-positive neurons. Caspase-3 expression was determined by immunohistochemical analysis in sections 0-5.00 mm rostral to the epicenter at 6,12,24 hours, 3 and 7 days after injury. Results Neurons continued to disappear over 7 days after injury and neuronal loss extended to the areas far away from the epicenter over times except 0-0.50mm. TUNEL-positive and Caspase-3 positive neurons in gray matter appeared after 6 hours, gradually increased to a peak level after 3 days at 2.55mm and 3.10mm, and declined by 7 days postinjury in the section 1.10-4.60mm rostral to the epicenter. Conclusion Temporal and spatial distribution of neurons apoptosis and Caspase-3 protein expression after spinal cord injury are regulated. References | Related Articles | Metrics

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Activated astrocyte secrete clusterin that enhances neuronal differentiation from rat hippocampal neural stem cells ZOU Lin-qing JIN Guo-hua* LI Hao-ming QIN Jian-bing TIAN Mei-ling ZHU Pei-pei WANG Hai-qin 2013, 44 (6 ):  761-766.  doi: 10.3969/j.issn.0529-1356.2013.06.006 Abstract ( )   Objective To detect the secretion of clusterin in "reactive astrocytes" which are activated by fimbria-fornix transected hippocampal extracts, and to investigate the neuronal differentaition of hippocampal neural stem cells (NSCs) by clusterinin vitro. Methods The astrocytes were cultured in fimbria-fornix transected hippocampal extracts, normal hippocampal extracts and DMEM/F12 medium, the conditional medium contained normal hippocampal extracts and transected 12hours, 24hours, 48hours hippocampal extracts. The concentrations of clusterin was detected by ELISA assay. To investigate the neuronal differentaition of hippocampal neural stem cells (NSCs) by clusterin, the cells were divided into the induced group and control group. Seven days later, the neuronal differentiation of hippocampal NSCs was investigated by MAP-2 immunofluorescence and labeled by Hoechst. Results The concentrations of clusterin in normal group and control group were lower than the transected 12hours, 24hours, 48hours groups, especially the 24hours group which concentration was about 4-folds higher than normal and control group，about (0.1037±0.0119)ng/L,(0.1170±0.0078) ng/L, (0.1940±0.0364) ng/L, (0.4250±0.0724) ng/L and (0.2903±0.0472) ng/L, respectively. The number of MAP-2 positive cells in clusterin induced group was higher than the control group (P<0.05), and the percentages of the total number of cells of these two groups were (11.67± 2.57)% and (4.52±1.54)%, respectively. The processes in the induced group was longer and more than the control group. Conclusion Reactive astrocytes activated by fimbria-fornix transected hippocampal extracts secrete more clusterin. The clusterin in culture cells may promote hippocampal NSCs differentiation into mature neurons in vitro. References | Related Articles | Metrics

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Changes of RUNX1T1 mRNA and protein expression in rat hippocampus after fimbria/fornix transection ZOU Lin-qing JIN Guo-hua* WANG Hai-qin LI Hao-ming ZHU Pei-pei QIN Jian-bing 2013, 44 (6 ):  767-771.  doi: 10.3969/j.issn.0529-1356.2013.06.007 Abstract ( )   Objective To observe the temporal and spatial changes of RUNX1T1 mRNA and protein expression in rat hippocampus after fimbria/fornix transection at different time points. Methods Rat models were prepared by cutting fimbria fornix, and divided into normal group, cutting 3days, 7days, 14days, 21days and 28days groups. 1. The total hippocampal tissue mRNA was extracted, and the expression of RUNX1T1 mRNA was detected by real-time PCR; 2. The total protein of hippocampus was extracted, and the expression of RUNX1T1 protein was detected by western blotting; 3. Frozen brain sections were detected by RUNX1T1 immunofluorescence and Hoechst labeled, to observe the RUNX1T1/Hoechst double labeled positive cells in hippocampal dentate gyrus. Results Real-time PCR analysis showed that the RUNX1T1 mRNA expression in the hippocampus was significantly upregulated on the 3rd day after transection. Western blotting analysis indicated that the expression of RUNX1T1 protein was detected in the hippocampus of rats at basal condation, and the levels of RUNX1T1 protein were markedly increased at days 3 and 7, and peaked at day 3 after transection. In contrast, no siginificant change in the RUNX1T1 protein expression at days 14, 21 and 28. Immunostaining showed that the nember of RUNX1T1-positive cells was siginificantly higher at day 3 compared with control and decreased slowly after day 7. Conclusion The transient upregulation of RUNX1T1 mRNA and protein expression in the early phase after fimbria/fornix transection, and location in the hippocampal dentate gyrus granule lower, which may be related to the hippocampal neural regeneration. References | Related Articles | Metrics

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Effects of ethanol on autophagy and the role of P62 in ethanol-induced autophagy in pheochromocytoma cells DONG Yue-juan SHI Zhen-yu LIU Shi-meng ZHANG Wei-juan HUANGFU Chao-shen DENG Jin-bo LIU Bin* 2013, 44 (6 ):  772-777.  doi: 10.3969/j.issn.0529-1356.2013.06.008 Abstract ( )   Objective To utilize pheochromocytoma (PC12) cells to examine the effect of ethanol on autophagy and the role of P62 in ethanol-induced autophagy. Methods The inhibition effect of ethanol on proliferation of PC12 cells was examined by MTT assay. Indirect immunofluorescence was used to detect location of LC3, a biomarker of autophagy, and P62. The LC3 fluorescence intensity in cellular cytosol was measured using a high content screening imaging system. The ultrastructural morphology of autophagic vacuoles and autophagy-lysosome was observed under the transmission electron microscope. Protein expression of P62 was detected with Western blotting analysis. Results Compared to the control in serumfree medium after 24hours, ethanol at the concentrations of 100mmol/L, 200mmol/L, 400mmol/L and 800mmol/L decreased cell proliferation of PC12 cells to 91.97% (P<0.05)，72.63%(P<001)，58.23%(P<0.01) and 50.82%(P<0.01) respectively, indicating a dose-dependent inhibitory effect of ethanol on cell proliferation.Indirect immunofluorescence staining and the high content screening imaging system showed that ethanol increased LC3 fluorescence intensity and the co-localization of LC3 with P62 in PC12 cellular cytosol. The transmission electron microscope showed the ultrastructural morphology of autophagic vacuoles and autophagy-lysosome in PC12 cells incubated for 2hours in the presence of 200mmol/L ethanol. Compared with the control in serum-free medium, the protein expression of P62 increased significantly in different ethanol concentration treatment groups (P<0.01). The crest-time appeared at 2hours after the PC12 cells were treated with 200mmol/L ethanol. Conclusion Ethanol may induce autophagy of PC12 cells. P62 may be involved in the autophagy against ethanol-induced cytotoxicity. References | Related Articles | Metrics

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Cloning and expression pattern of heat shock protein 90 gene from planarian Dugesia japonica MA Ke-xue FENG Cheng-cheng DOU He CHENG Fang-fang CHEN Guang-wen* LIU De-zeng 2013, 44 (6 ):  778-783.  doi: 10.3969/j.issn.0529-1356.2013.05.009 Abstract ( )   Objective To clone heat shock protein 90 gene from planarianDugesia japonica (Djhsp90), and to examine its expression pattern in response to different stressors. Methods The full-length cDNA and genomic DNA of Djhsp90 was cloned using PCR methods, and its sequence was analyzed by the bioinformatics software, and its expression pattern in response to different stressors was detected by the fluorescent Real-time RT PCR. Results The full-length cDNA of Djhsp90 was 2354 bp containing an open reading frame (ORF) of 2148 bp, which encoded a polypeptide of 715 amino acids with five signatures motifs and the C-terminal consensus MEEVD of eukaryotic HSP90 family. The ORF sequence of genomic DNA was sequenced, and found that only one intron (48 bp) existed in Djhsp90 gene structure. The 5’ and 3’ putative splicing site followed the typical “GT-AG” rule. Djhsp90 expression began to rise at 1 day following the amputation of planarians, reached a peak at day 2 and decreased gradually at day 3 to the normal level. During the prolonged starvation, Djhsp90 expression remained a relative higher level in contrast to the controls. We verified that elevating the culture temperature strongly induced Djhsp90 expression, and the most sensitive induction occurred in the head fragment of planarians. Conclusion We have successfully cloned hsp90 gene from planarianDugesia japonica, and verified that amputation, starvation and thermal stress can induce the up-regulation of Djhsp90. References | Related Articles | Metrics

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Comparative ound analysis effects of separated cardiomyocytes by collagenase Ⅰ and Ⅱ WANG Hui-ling GUO Kang GUO Zhi-kun* 2013, 44 (6 ):  784-788.  doi: 10.3969/j.issn.0529-1356.2013.06.010 Abstract ( )    Objective To explore the role of collagenase Ⅱ in the separation of adult rat cardiomyocytes. Methods Collagenase Ⅰ and collagenase Ⅱ were used to digest cardiomyocytes and their effect of separating cardiomyocytes was compared. Immunohistochemistry, immunofluorecence and Western blotting were used to detect existence of collagen Ⅱ in adult rat myocardium. Collagen Ⅱ of xiphoid and collagen Ⅲ of myocardium were taken as positive control. SDS-PAGE(polyacrylamide gel electrophoresis) method was used to detect the composition of collagenase Ⅱ. Results After digesting cardiomyocyte with collagenase Ⅰ or collagenase Ⅱ, the number of the obtained myocardial cells were (5.66±0.83)×106 and (5.61±1.01) ×106, the survival rates of cardiomyocyte were（74.17±6.27）％and（75.50±9.07）%, respectively, and no significant difference was found. Most separated cardiocytes did not beat while only very few of them had automaticity systole; Collagen Ⅱ showed negative expression in myocardium of adult rats but positive expression in cartilaginous tissue of xiphoid; Collagen Ⅲ also showed positive expression in myocardium. Western blotting did not show the expression of collagen Ⅱ in myocardium. SDS-PAGE results showed collagenase Ⅱ was a mixture of five kinds of protein. Conclusion The collagen Ⅱ did not exist in myocardium, thus collagenase Ⅱ should not be used to separate adult rat cardiomyocyte. References | Related Articles | Metrics

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Changes of vimentin and matrix metalloproteinases-2 expressions in epidermal grouth factor induced migration of human umbilical vein endothelial cells LU Shi-jun YANG Hui-ke MA Wen-ping LI Xiao-dong ZHANG Ya-fang* 2013, 44 (6 ):  789-794.  doi: 10.3969/j.issn.0529-1356.2013.06.011 Abstract ( )   Objective To evaluate the relationship between vimentin and migration induced by epidermal growth factor (EGF) in human umbilical vein endothelial cells (HUVECs), the expressions of vimentin and matrix metalloproteinases-2 (MMP-2) were examined in epidermal growth factor (EGF) -induced migration of HUVECs. Methods In the current study, the migration of HUVECs was analyzed after EGF stimulation for 10h by scratch assay, and the effect of EGF on the cells morphology was also observed. The monolayer HUVECs were scratched to induce the cells migration in both control group and EGF group, and then immunofluorescent staining was used to detect the vimentin expression. Both vimentin and MMP-2 expressions were assessed via Western blotting and RT-PCR. Results The scratch assay revealed a 1.8-fold increase induced by EGF in migration levels of HUVECs. Interestingly, the HUVECs morphology also altered dramatically from cobblestone-like shape to spindle-like shape after EGF stimulation. As a result of EGF stimulation, both vimentin and MMP-2 expressions in HUVECs were up regulated in both protein and mRNA levels (P<0.05). Conclusion EGF upregulates both vimentin and MMP-2 expressions effectively in EGF-induced migration of HUVECs, and vimentin is closely associated with the migration in HUVECs. References | Related Articles | Metrics

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Expression of islet amyloid polypeptide and glucose transporter 2 in the islet β cell of rat during heroin withdrawal, detoxification and relapse HAN Jing LIANG Wen-mei* HONG Yan XIA Bai-juan HU Yun LI Yi-xin 2013, 44 (6 ):  795-799.  doi: 10.3969/j.issn.0529-1356.2013.06.012 Abstract ( )   objects To investigate the expression changes of IAPP and GLUT2 in islet βcells during heroin withdrawal induced by naloxone, methadone detoxification treatment, and heroin relapse. Methods Male Sprague-Dawley rats were divided into normal control group (NCG) and experiment group (EG). The EG was further divided into heroin withdrawal group (HWG), methadone detoxication group (MDG), and heroin relapse group (HRG). Pancreas was drawn from each group. Immunohistochemistry and image analysis were used to detect the expression of IAPP and GLUT2. Results As compared with the normal control, the immunostainning of IAPP and GLUT2 turned express intensive inβcells of islets from heroin withdrawal and relapse rats than that in controls; the numerical density on area of IAPP also increased; the mean gray values decreased in these two groups. There is no difference were found in the immunostainning, the mean gray values and the numerical density on area during detoxification treatment compared to controls. Conclusion During heroin withdrawal and relapse, the expression increased. While in detoxification, they became decreased. The changes of IAPP and GLUT2 are consistent and had same rhythm in three stages. It showed that IAPP and GLUT2 take participate in energy metabolic process, , which may be related to food intake and body weight, or protective function toβcells. References | Related Articles | Metrics

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Relationship between the posterior-lateral corner of femoral insertion and external opening of femoral tunnel used in reconstruction of the anterior cruciate ligament DONG Yi-long* CAI Chun-yuan JIANG Gang-yi QIAN Yue-nan JIANG Wen-hui ZHAO Zhang-wei YANG Guo-jing 2013, 44 (6 ):  800-803.  doi: 10.3969/j.issn.0529-1356.2013.06.013 Abstract ( )   Objective To locatize the positions of posterior-lateral corner(PLC) of femoral insertion and external opening of femoral tunnel used in reconstructure of the anterior cruciate ligament (ACL) in order to provide anatomical data for one-stage ACL and PCL anatomical reconstruction. Methods We used 30 adult cadaveric knee specimens. With flextion of knee at 120°, the anatomical ACL femoral tunnel reconstruction was performed under arthroscopy through anteromedial portal and marked with Kirschner. We identified femoral insertion site of the lateral collateral ligament(LCL) and popliteus tendon(PT) on the femoral condyle of the knee. We used the femoral epicondyle as the origin and established(x, y) vertical axis to measure the coordinate of femoral anatomical insertion site of LCL, PT and external opening of ACL femoral tunnel. We measured distances between these three points. Results The LCL insertion site was located at a mean distance of (1.27±3.35)mm proximally and (2.99±1.29)mm posteriorly, to the lateral epicondyle. The PT insertion site was located at a mean distance of (8.85±3.38)mm disstally and (3.83±1.95)mm posteriorly, to the lateral epicondyle. The external opening of the ACL femoral tunnel was located at a mean distance of (16.12±5.34)mm proximally and (6.84±4.17)mm posteriorly, to the lateral epicondyle. The distance between LCL and PT insertion site was (9.67±3.92)mm. The distance from the external opening of ACL femoral tunnel to LCL insertion site was (13.07±4.93)mm, and to PT insertion site was (23.37±6.16)mm. Conclusion We reveal anatomic characteristic of femoral insertion site of the LCL and PT and external opening of ACL femoral tunnel, and provide anatomic basis for one-stage clinical combination reconstruction. References | Related Articles | Metrics

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Association of morphogenesis of pulmonary endoderm with development of prepharyngeal mesenchyme and outflow tract septation in mouse embryos LI Hui-chao JING Ya* SHI Liang YANG Yan-ping LI Hai-rong QIAO Cong-jin QIAO Ai-xiu CAI Yu-jin 2013, 44 (6 ):  804-811.  doi: 10.3969/j.issn.0529-1356.2013.06.014 Abstract ( )   Objective To explore the relationship of pulmonary endoderm with the development of prepharyngeal mesenchyme and the outflow tract septation of the mouse embryonic heart. Methods Both the immunohistochemical and immunofluorescent methods were used in the present studies in order to observe islet-1(ISL-1), α-smooth muscle actin(α-SMA), sonic hedgehog(Shh); patched(Ptc1), patched 2(Ptc2), smoothened(Smo) and myosin heavy chain(MHC) distribution in 45 mice embryos from embryonic day(ED)8 to embryonic day(ED)13. Results From ED8 to ED9, ISL-1 positive cells distributed in the splanchnic mesoderm of the pericardial cavity dorsal wall, bilateral mesenchyme to the foregut, and extended distally to the primary heart tube. Ptc1, Ptc2 positive cells were detected in the myocardium of the heart tube. During ED10 to ED13, developing pulmonary endoderm extended ventrally and expressed Ptc1, Ptc2 strongly. ISL-1 positive prepharyngeal mesenchyme surrounding the pulmonary endoderm formed a symmetrically distinctive cone-shaped structure, which projected into the aortic sac to form the primary aorta-pulmonary septum. At ED12, most of the cells in aorta-pulmonary septum turned to express α-SMA, only a few ISL-1 positive cells could be detected. Conclusion The differentiation and development of pulmonary endoderm are closely associated with typical aggregation of ISL-1 positive prepharyngeal mesenchyme. The developing pulmonary endoderm probably acts as an organizing center via sonic hedgehog signaling pathway to confer positional information on ISL-1 positive cells aggregation. The ventral extension of the pulmonary endoderm may not only induce the accumulation and migration of ISL-1 positive cells, but also play important roles in normal morphogenesis of the outflow tract and establishment of effective pulmonary circulation. References | Related Articles | Metrics

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Effects of rabbit antiserum on mouse embryonic stem cells at 8-cell embryo development stage HU Chun-chao CUI Li-fang WANG Peng-bo WANG Mian-juan DI Ke-qian LI Xiang-yun* 2013, 44 (6 ):  812-818.  doi: 10.3969/j.issn.0529-1356.2013.06.015 Abstract ( )   Objective Mouse 8-cell embryos were cultured in KSOM media with rabbit antiserum to mouse embryonic stem (ES) cells, and the effect of the antiserum on mouse embryos was observed. Methods The antiserum was diluted to 1/50, 1/100 and 1/200 by KSOM medium, respectively. Results The result of morphology changes showed that the embryonic development was inhibited or even stopped in the 1/50 dilution group, while the compaction was blocked, the embryos showed fluid vacuoles in the blastula stage in the 1/100 dilution group; there was no obvious effect in the 1/200 dilution group. The treatment group with normal rabbit serum did not appear differences from the control group. Our data indicated that the compaction was blocked by antiserum; when the compacted embryos were exposed in the antiserum, decompaction occurred; prolonged decompaction beyond the 32-cell embryo resulted in an increasing proportion of malformed blastocysts. The embryo development was further tested in the 1/100 dilution group by compaction rate, blastocyst rate, inner cell mass (ICM) outgrowth rate, cell counting, embryo transplantation and fluorescent staining. The result showed that the 1/100 dilution antiserum inhibited ICM formation, decreased embryonic cell number, and creased abnormal blastocyst rate. The treated blastocysts were transferred into uterine horns and did not develop to term. Conclusion Our data suggests that the rabbit antiserum to mouse ES cells inhibits embryo compaction, ICM formation and postimplantation development. References | Related Articles | Metrics

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Effects of extract of notopterygium on Th1/Th2 cells balance and p38 signaling pathway in the asthma mouse model LI Liang-chang PIAO Hong-mei QIN xiang-zheng YAN Guang-hai LI Guang-zhao* 2013, 44 (6 ):  819-823.  doi: 10.3969/j.issn.0529-1356.2013.06.016 Abstract ( )   Objective To explore the effect of extract of notopterygium (EN) on Th1/Th2 cells balance and signaling pathway p38 in the asthma mouse model. Methods Sixty male BABL/c mice were randomly divided into six groups with 10 mice per group: normal group, ovalbumin（OVA）group, notopterygium low dose group, notopterygium high dose group, SB203580 group and dexamethasone group. The left lung was isolated for pathological examination. Lung sections were stained with HE. The number of total cells and different inflammatory cells were counted by an optical microscope. The concentrations of interleukin(IL)-4，IL-5，IL-13 and interferon-γ(IFN-γ) in bronchoalveolar lavage fluid (BALF) were measured by ELISA. The Western blotting was performed to detect the IL-4, IFN-γ， P38 in lung tissue. Results In OVA group, the number of total cells and inflammatory cells, the concentrations of IL-4, IL-5, IL-13 in BALF were significantly higher than those in normal group (P<0.05), but the concentration of IFN-γ in BALF was significantly lower than in normal group(P<0.05). Pulmonary inflammatory infiltration was more serious in OVA group than in normal group(P<0.05). IL-4 and phosphorylated P38 in lung tissue were significantly higher in OVA group than those in normal group (P<0.05), but IFN-γ level of lung tissue was significantly lower than in normal group (P<0.05). In notopterygium low and high dose groups, SB20358 group and dexamethasone group, the number of total cells and inflammatory cells, the concentrations of IL-4, IL-5, IL-13 in BALF and IL-4, phosphorylated P38 in lung tissue were significantly lower than those in OVA group (P<0.05), the level of IFN-γ in BALF and lung tissue was significantly higher than in OVA group (P<0.05); Notopterygium, SB20358 and dexamethasone significantly reduced pulmonary inflammatory infiltration(P<0.05). Conclusion All these results indicate the extract of notopterygium roots can inhibit the development of asthma in the mouse model, and its possible mechanism is that Th1/Th2 cells balance may be changed while p38 signaling pathway is inhibited. References | Related Articles | Metrics

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Physical characteristics of the Han ethnic groups of Mindong dialect in Fujian YU Ke-li ZHENG Lian-bin* HU Ying WANG Yang XUE Hong CHENG Zhi DENG Wei SHI Rui 2013, 44 (6 ):  824-834.  doi: 10.3969/j.issn.0529-1356.2013.06.017 Abstract ( )   Objective To analyze the physical characteristics of Han with Mindong Dialect. Methods Investigation in strict accordance with the methods of Martin,Anthropometric Methods andAnthropomorphic Handbook, eighty-six physical characteristics on 692 adults (151 urban males, 188 rural males, 159 urban females and 194 rural females) of Han with Mindong dialect were investigated in Fujian province. Twenty-six physical indices were calculated and the distributions of indices were done. Comparison with the data of ethnic groups in China, and preliminarily. Results The percentage of eyefold of the upper eyelid was high. The percentage of mongoloid fold was low. Opening height of eyeslits was narrow. External angles were higher than internal angles in direction of eyeslits. The nasal profile was straight. The height of alae nasi was middle-type. The nasal base was upturned. The maximal diameter of nostrils was oblique. The breadth of alae nasi was wide type. The zygomatic projection was tiny. The upper lip skin height was middle type. Thicknesses of lips were mainly thin. Hair color was black and skin color was yellowish. The eye color was brown. Urban males, rural males and urban females were all super middle-sized stature, while rural females were middle-sized stature. According to the mean of headface and body indices, males were brachycephaly, hypsicephalic type, metriocephalic type, leptoprosopy, leptorrhiny, mesatiskelic type, medium chest circumference, narrow distance between iliac crests, long trunk and broad shoulder breadth. In addition, females were mesocephaly, hypsicephalic type, aerocephalic type, hyperleptoprosopy, leptorrhiny, mesatiskelic type, medium chest circumference, narrow distance between iliac crests and middle trunk. And urban females were medium shoulder breadth, while rural females were broad shoulder breadth. Characteristics of head and face of Han with Mindong dialect were affected by North Asian ethnic groups and South Asian ethnic groups, while body indices were close to North Asian ethnic groups. Conclusion The physical characteristics of Mindong Han are relatively close to the North Asian type ethnic groups. References | Related Articles | Metrics

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Physical characteristics of Tunbu YU Ke-li ZHENG Lian-bin* BAO Jin-ping LI Yong-lan RONG Wen-guo QI Xiao-lin FENG Chen-lu NI Xiao-lu 2013, 44 (6 ):  835-842.  doi: 10.3969/j.issn.0529-1356.2013.06.018 Abstract ( )   Objective To investigate the physical characteristics of Tunbu. Methods Eighty-seven physical characteristics of 507 Tunbu adults (251 male, 256 female) from Anshun, Guizhou province were investigated. Twenty-three body mass indexes were calculated, statistics on index typing was performed, and the physical characteristics of Tunbu were analyzed preliminarily with the comparison to our ethnic data. The investigation was carried out in accordance with the methods from Martin,Anthropometric Methods andAnthropometric Manual. Results Most of Tunbu had eyelid folds, and 1/5 adults had Mongolian folds. Most of their paropia were high; most of their nasion heights were medium with straight bridges; most of their nasal bases were level; most of their alae nasi height were middle; the rates of oblique and transverse ala nasi of the maximum nostrils diameter were relatively closed each other; the rate of round earlobe was the highest; the height of upper lip skin was mostly moderate; most of lips were moderate; most of their hair was black; skin was mostly yellowish; eyes were mostly brown. Heights of Tunbu men and women were sub-medium. Male and female of Tunbu were mostly mesocephaly, hypsicephalic type, aerocephalic type, mesorrhiny, long trunk, and broad distance between iliac crests. Males were mostly euryprosopy, mesatiskelic type, medium chest circumference, medium shoulder breadth and short type; Females were mostly broad chest circumference. The rate of euryprosopy and mesoprosopy, subbrachyskelic type and mesatiskelic type, narrow shoulder breadth and medium shoulder breadth, short and sub-middle were relatively similar. Conclusion The physical characteristics of Tunbu are close to Han in other areas and North Asian ethnic groups. References | Related Articles | Metrics

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Physical characteristics in head-face of Tujia ethnicity children and adolescents HUANG Da-yuan* ZHANG Hui-juan SHI Hui-juan WU Guo-yun ZHU Yao-feng 2013, 44 (6 ):  843-850.  doi: 10.3969/j.issn.0529-1356.2013.06.019 Abstract ( )   Objective To explore the physical characteristics and patterns in head-face of Tujia ethnicity children and adolescents. Methods According to the anthropometric method, eleven head-face characteristics of 1865 Tujia children and adolescents（953 males and 912 females）in Wuling Mountain area were investigated, and thirteen physical indices were calculated by formula. Results The mean values of measurements gradually increased with age, and there were significant differences between males and females in each age group.Male heads were wider and shorter than female heads, and their faces were narrower and longer than female faces. Head shapes of Tujia children and adolescents were characterized by brachycephaly, orthocephalic and tapeinocephalic, and their facial types were mainly featured by euryprosopy and euryen. Conclusion The physical development in head-face of Tujia children and adolescents is in conformity with the general growth pattern. Difference of head-face characters between male and female, and difference of head-face characters between Tujia and Dong of Guangxi and Miao of Guangxi students are significant. References | Related Articles | Metrics

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Facial characteristics of Man nationality in Sanjiazi village of Heilongjiang province WEN You-feng* YANG Yang WANG Xia XI Huan-jiu 2013, 44 (6 ):  851-855.  doi: 10.3969/j.issn.0529-1356.2013.06.020 Abstract ( )   Objective To investigate the facial characteristics of Man nationality adults living in Sanjiazi village of Heilongjiang province. Methods With the informed consent, according to the human body measurement method, a study on somatoscopy for eyefold of upper eyelid, lobe types and 18 head and facial index was carried out in 134 Man nationality adults(62 males and 72 females) living in Sanjiazi Village of Heilongjiang province. The obtained data were input into SPSS16.0 software package for statistics processing, like frequency analysis, chi-square test, etc. Results There were significant gender differences in nasal root height, nasal profile, nose base department, nostrils type, eyefold of upper eyelid, chin, forehead hair point, hair color and eye color according to head and facial investigate index of Man nationality adults in Sanjiazi Village of Heilongjiang province. There were some common facial characteristics among them: flat or moderate zygomatic projection, having mongolodid wrinkle, the secondary palpebral fissure height. The direction of eye slits was exterior angle higher than interior angle. The height of wing nose and breadth of nasal ala were mediumsized. The largest footpath position of nostril was oblique. The earlobe type was evenly distributed. The hair form was the straight type. Conclusion The Man nationality in Sanjiazi village of Heilongjiang province belongs to the East Asian type of the Mongoloid race with the physical characteristics of Northern Chinese. References | Related Articles | Metrics

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Body morphological traits of Oroqen adults in Heilongjiang province WEN You-feng* WANG Xia YANG Yang CAO Fang XI Huan-jiu 2013, 44 (6 ):  856-860.  doi: 10.3969/j.issn.0529-1356.2013.06.021 Abstract ( )   Objective To investigate the body morphological traits of Oroqen adults in Heilongjiang province. Methods On the base of the informed consent, 157 Oroqen adults(male 75, female 82) from Tahe County and Huma nationality village, Heilongjiang province were randomly selected for the study. Followed the somatometric measurements, we measured thirty-four traits about trunk and limbs including shoulder tip height, suprasternal height and so on, calculated nine classificational indices of the body. Results There were significant differences in body features about Heilongjiang Oroqen between males and females, except pelvic breadth and chest depth. Males were higher, females were shorter. Both males and females belonged to wide shoulder type, long trunk type, wide pelvis type, hyperbrachyskelic. Conclusion Compared with Inner Mongolia Oroqen, the leg of Oroqen in Heilongjiang is shorter, pelvis wider, shoulder wider.These differences may be related to the local climate. References | Related Articles | Metrics

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Isolation and culture of first trimester human trophoblast cell ZUO Yan-zhen ZHANG Kong-yan LI Yu-hong* XU Qian MAO Xiao-dan TAN Yu-si 2013, 44 (6 ):  861-864.  doi: 10.3969/j.issn.0529-1356.2013.06.022 Abstract ( )   Objective To research a simple method for purification of first trimester human trophoblast cell in vitro.Methods Whole first trimester human chorionic tissues were isolated with trypsin/collagenase. HE staining and immunocytochemistry methods were used to detect the tyophoblast cells purity. Results Cytokeratin-7 positive cells were more than 90%. Conclusion First trimester human Trophoblast cells can be obtained through the simple method and may offer enough cells for later experiments. References | Related Articles | Metrics

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Comparison of protein preparation methods for Gel-based proteomics BU Wang-yu QI Yi-jun* WEI Hua ZHANG Juan MA Jin WANG Ming MA Yuan-fang 2013, 44 (6 ):  865-868.  doi: 10.3969/j.issn.0529-1356.2013.06.023 Abstract ( )   Objective To evaluate the efficiency of protein preparation for human esophageal tissue protein diluted in PBS by ultracentrifugation and 7 protocols for protein precipitation. Methods Eight protocols, which included ultracentrifugation and 7 protocols for protein precipitation ［ammonium sulfate, chloroform/methanol, acidified acetone/methanol, ethanol, acetone, trichloroacetic acid (TCA), TCA/acetone］, were used to extract PBS-diluted mixed protein samples derived from human esophageal cancer tissue (10 cases). Protein concentration was determined by bicinchoninic acid(BCA), Bradford and 2D quantification kits. The recovered protein diluted in 2D rehydration buffer was separated either by one-dimensinal mini-gel electrophoresis or two-dimensional gel electrophoresis (2D) followed by ImageMaster software analysis. Results Protein quantification protocols by Bradford and 2D were compatible with 2D rehydration buffer. With the exception of the protein recovery rates for the ultracentrifugation and ammonium sulphate precipitation which were 40% and 4%, respectively, the protein recovery rates for remained methods were 22%. In view of protein spot boundary, number and resolving ability, aectone-precipitated protein produced the best quality of 2D images, followed by precipitation protocols of ethanol and chloroform/methanol, and ultracentrifugation. Acidified acetone/methanol protocol produced the greatest protein spot lost. Conclusion Acetone precipitation protocol is feasible with protein samples with high concentration and small volume, while ultracentrifugation is for other protein samples. References | Related Articles | Metrics

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Circulating fibrocytes and organic fibrosis FU Xu-yan LI Yu* LI Ya-dong NIU Jian-zhao 2013, 44 (6 ):  869-872.  doi: 10.3969/j.issn.0529-1356.2013.06.024 Abstract ( )   Fibrosis of organs is related with over-accumulation of extracellular matrix(ECM). It is widely acknowledged that (myo)fibroblasts lead to the formation of extracellular matrix. Recent studies confirm that marrow-derived circulating fiobrocytes play important roles in the formation and process of fibrosis. Activated circulating fibrocytes not only secrete extracellular matrix that is essential for the process of fibrosis, but also differentiated into (myo)fibroblasts leading to fibrosis, either physiologically or pathologically. Circulating fibrocytes have special characteristics and various bio-functions, and secrete various cytokines to accelerate the fibrosis. The formation, migration, differentiation and fibrosis-promoting function are also regulated by various cytokines and pathways. This article aims to review the latest development in circulating fibrocytes and organic fibrosis. References | Related Articles | Metrics