Acta Anatomica Sinica

The pathological changes of the olfactory bulbs in the Niemann-pick type C1 mice

Xin YAN Liang QIAO Lin Juntang

2016, 47 (1): 2-6.

Abstract ( )

Objective To examine the impact of Niemann-pick type C1 disease on the olfactory system, we investigated the morphological changes of the olfactory bulbs in NPC1 mutant mouse. Methods The genomic DNA was extracted from mice tails for genotyping by PCR；30-day-old mice including wild type and NPC1 gene mutation were selected, and immunohistochemistry was performed to examine and compare the axonal structure, the myelination process, as well as the expression of SMI31 and MBP in the olfactory bulbs；The cell apoptosis were analyzed by TUNEL assay. Results In NPC1-/- mice, the expression of NPC1 protein is rarely detected, but TUNEL-positive cells are increased. Immunofluorescence using SMI31 antibody indicates accumulation of neurofilament and formation of axonal spheroids in the NPC1-/- olfactory bulbs. Furthermore, the decreased expression of MBP protein is also found in NPC1-/- olfactory system, which is associated with perturbed myelination. Conclusion Our data show pathological changes in the olfactory bulbs of NPC1 mutant mice, which is accompanied by olfactory defects.

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Ultrastructural features of enteric glial cells and the distribution of S-100 beta in the small intestine of chicken

2016, 47 (1): 7-11.

Abstract ( )

objective To observe the structure of Enteric glial cell and distribution of S-100β protein in 10 healthy yellow feather broiler chickens. Methods The ultrastructural characteristics of glial cells were observed by transmission electron microscopy， and the distribution of S-100 protein was studied by immunohistochemical SABC-AP method. Results The electron microscope showed that chicken enteric glial cells (EGCS) in the small intestine were star， irregular nucleus， cytoplasm distribution has a small round or oval unmyelinated nerve fibers. Immunohistochemistry showed that in chicken intestine epithelium mucosa of each segment， intestinal glands of S-100βwere expressed strong. Among the basement membrane of epithelial cells and at the top of intestinal gland epithelial cells were the strong positive， lamina propria weaker expression， in the submucosal plexus and myenteric plexus showed strong positive expression. Conclusion the glial cell ultrastructure characteristics and distribution characteristics of S-100βare similar to the small intestine of mammals.

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Effect of resveratrol on expression of synaptophysin after cerebral ischemia/reperfusion injury in rats

2016, 47 (1): 12-17.

Abstract ( )

[Abstract] Objective To examinate the effect of resveratrol on expression of synaptophysin after cerebral ischemia/reperfusion injury in rats. Methods 60 male Sprague-Dawley rats were divided into sham, ischemic control and resveratrol group. Each group is 20 rats. The rats were treated with resveratrol or ethyl alcohol by intraperitoneally for 7 consecutive days after 3 h for MCAO. At 24 h after MCAO/R, TTC assay examinated cerebral infarction volume and dry and wet weight method tested moisture content in brain tissue. At 24 h, 7 d and 14 d after MCAO/R, we examinated the neurological function score with Bederson score and the protein expression of synaptophysin in ischemic cortex with immunohistochemistry and Western blot. Results The infarction volumes in the control and resveratrol group were significantly higher than in the sham group (P < 0. 05), and it was significantly lower in the resveratrol group than in the control group (P < 0. 05).Brain water contents in the control and resveratrol group were significantly higher than in the sham group (P < 0. 05), and it was significantly lower in the resveratrol group than in the control group (P < 0. 05). The neurological function score in the control and resveratrol group were significantly higher than in the sham group at 24 h, 7 d and 14 d (P < 0. 05), and it was significantly lower in the resveratrol group than in the control group (P < 0. 05). Immunohistochemistry display that the positive expression of synaptophysin in ischemic cortex in the control group were reduced at 24 h, and increased gradually at 7 d and 14 d, but were significantly lower than in the sham group. After resveratrol treatment, it was significantly higher than in the control group at each point time, and was highest at 14 d. Western blot assay showed that the protein expression of synaptophysin in the control group was significantly lower than in the sham group at 24 h, 7 d and 14 d after MCAO/R (P < 0. 05), but gradually increased at 7 d and 14 d. After resveratrol treatment, it was significantly enhanced than in the control group at each point time (P < 0. 05), and was strongest at 14 d. Conclusion Resveratrol treatment can enhance the expression of synaptophysin and improve neurological function.

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Induction and Differentiation of Neural Precursor Cells from IPSCs

2016, 47 (1): 18-22.

Abstract ( )

The induced pluripotent stem cells (iPSCs) are expected to be candidate cells for cell transplantation in treatment of ischemic brain injury since they have a strong ability to proliferate and differentiate, and an easy accessibility without immune rejection and ethical issues. However, recent experiments have found that the iPSCs transplanted into the brain almost form tumors. Inducing the iPSCs into neural precursor cells (NPCs) in vitro before tansplantation may avoid tumor formation in the brain. The present study therefore has used the N2B27 as medium in a multistep adherent culture to induce the iPSCs into the NPCs. Immunofluorescence staining showed that the neural stem cell marker nestin was positive in the NPCs, in which there were very fewβ-Tub-III positive cells. After 7 days culture in the differentiation culture medium, β-Tub-III positive cells increased. This experiment has confiemrd that the iPSCs can be induced into the NPCs in vitro, which are expected to be used in the treatment of ischemic brain injury.

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Histological differences of hippocampal formation in different Reelin genotype mice

YANG Ci-Qing

2016, 47 (1): 23-27.

Abstract ( )

Objective To provide scientific basis data for the function of Reelin in the hippocampal formation by comparing the histological structure of the hippocampus in different reelin genotype mice. Methods Heterozygous reelin mice offspring were used, and genotype identification was performed after birth for 10 days. Experiment was divided into three groups including reelin+/+, reelin+/- and reelin-/-, and each group had 3 mice. After identification, the mice were perfused through heart by 4% paraformaldehyde and the whole brain wascollected. All samples were fixed by paraformaldehyde, sucrose dehydration and finally cut into frozen coronal slice. DAPI was used to stain nuclei, and fluorescence immunohistochemistry was performed to label Tbr-1, NeuN and GFAP specific expression cells, then fluorescence microscope was used for further observation and analysis; Results From the DAPI stained nuclei and Tbr-1, NeuN and GFAP labeled cell structure, it could be seen that reelin+/+ mice and reelin+/- mice in hippocampal area had similar structure, but in comparison with the reelin+/+ and reelin-/- mice, dentate gyrus cells of the reelin-/- type mouse spreaded to the gate area, the cell layer of hippocampal structure disordered, and GFAP labeled cells reduced; Conclusion Reelin has obvious effect on normal formation of mouse brain structure. Under the condition of reelin knock out mouse, the dentate gyrus area cells are dispersed, the cell layer of the hippocampus is disordered, and glial cells and fiber distribution both are reduced.

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Effects of gentiopicroside on Caspase-3, Bax and Bcl-2 in neonatal rat hippocampal nurons following oxygen-glucose deprivation and reperfusion injury

2016, 47 (1): 28-33.

Abstract ( )

Objective To study the anti-apoptosis effects of gentiopicroside in neonatal rat hippocampal neurons following oxygen-glucose deprivation and reperfusion injury and elucidated the related mechanisms. Methods The neonatal rat hippocampal neurons were pretreated with gentiopicroside 24h before exposed to oxygen-glucose deprivation and reperfusion injury were used as an in vitro model of ischemia and reperfusion. The neuron morphological change and apoptosis rate were evaluated by Hoechst 33342 staining. The nerve cell lactate dehydrogenase (LDH) leakage rate was detected by spectrophotometry. The expression levels of Caspase-3, Bax and Bcl-2 mRNA and protein were detected by RT-PCR and Western-blot assay respectively. Results Compared with the control group, oxygen-glucose deprivation and reperfusion injury model group could significantly enhance the apoptosis rate of nerve cells, increase the LDH leakage rate. Compared with the model group, pretreated with gentiopicroside (40, 20, 10mg/L) could significantly attenuated neuronal damage, with evidence of decreased neurons apoptosis rate and LDH leakage rate. Compare with the model group, pretreated with gentiopicroside (40 mg/L) could effectively down regulate the expression of Caspase-3 and Bax in mRNA and protein level, and up regulate the expression of Bcl-2 level. Conclusion Gentiopicroside preconditioning could prevent neurons from oxygen-glucose deprivation and reperfusion injury, and the mechanism may be mediated by up regulated Bcl-2 and down regulated Caspase-3 and Bax.

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Role of Wnt/β-catenin signaling activated by lithium chloride in the aging of hematopoietic stem/progenitor cell in mice

Jie-Yu XIA LI jing Yan-Yan ZHANG JIA daoyong Meng-si ZHANG Lin-Bo CHEN JING pengwei Xiong-Bin CHEN Lu WANG Wang yaping

2016, 47 (1): 34-41.

Abstract ( )

[Abstract] Objective To investigate the role of Wnt/β-catenin signaling activated by Lithium chloride in the aging of Sca-1 + hematopoietic stem/progenitor cell（Sca-1 + HSC/HPC）. Methods Sca-1+ HSC/HPC was isolated by magnetic cell sorting (MACS) and divided into three groups. The control group was routinely cultured; the Licl group was treated by Lithium chloride on the concentration of 10mmol/L; the aging group was added D-galactose on the concentration of 166mmol/L. All groups were cultured for 48h. The capability of colony formation was examined by CFU-Mix cultivation. The proliferation of Sca-1+ HSC/HPC was detected by CCK-8. The SA-β-Gal staining was used to investigate the aging of Sca-1+ HSC/HPC. The expression of β-catenin、GSK-3β、P53、P21 was measured by immunofluorescent staining and Western blotting. ELISA test was used to evaluate the level of 8 hydroxy deoxyguanosine (8-OH-dG). Results Compared with the control group, the proliferation of Sca-1+ HSC/HPC treated by Lithium chloride and D-galactose was blocked; the colony formation of CFU-Mix was decreased; the ratio of the SA-β-Gal staining positive Sca-1+ HSC/HPC was increased; the expression of β-catenin、P53、P21 was up-regulated and GSK-3β down-regulated ;the level of intracellular 8-OH-dG was raised. Conclusions The activation of Wnt/β-catenin signaling may lead to the aging of Sca-1+ HSC/HPC, and suggestion of Sca-1+ HSC/HPC aging may be caused by the cellular DNA oxidative damage and the up-regulation of P53/P21 pathway.

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Change of angiotensin-converting enzyme 2 expression in the macrophage foam cell induced by oxidized low density lipoprptein

2016, 47 (1): 42-48.

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Objective To investigate the change of ACE2 expression in the macrophage foam cell induced by ox-LDL. Methods Different concentrations of ox-LDL were used to induce macrophage cell RAW264.7 into foam cells, the intracellular lipid droplets amounts were observed by oil red O staining, the content of MDA was measured to identify oxidative damage, Real-Time PCR was used to detect ACE2 and MAS mRNA expression, the activity of SOD and the content of GSH were measured by chemocolorimetry. Results The macrophages became larger and rounder after treated by ox-LDL (20、40、60、80μg·ml-1) for 24h by oil red O staining, the amount of lipid droplets in the cytoplasm increased in a dose-dependent manner. The MDA content was increased significantly compare with the control group using 60μg·ml-1 ox-LDL treatment (P<0.01), Real-time PCR results showed that ACE2 and MAS mRNA expression in model group were decreased (P<0.05), and the SOD activity and the GSH content were also decreased (P<0.01 or P<0.05) significantly in model group. Conclusion Macrophage foam cell formation is closely related with the down-regulation of ACE2 and MAS expression, and that means ACE2 has anti-oxidative effect which may be related to SOD and GSH.

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Evaluation of cardiomyocyte differentiation of CD73 subpopulations in adipose derived mesenchymal stem cells

GUO ZHIKUN Zhang Min Gao Jianhui Li Qiong Yu Xia Wang Jingjing

2016, 47 (1): 49-54.

Abstract ( )

Objective The cardiac differentiated potency about subpopulations of CD73+/CD73- ADMSCs were evaluated for selecting subsets of cardiac differentiation advantage. Methods Using flow cytometry to separate CD73 +/CD73- ADMSCs subgroups, their shape were detected by HE staining. After ADMSCs subgroups induced by 5-aza, the expression of c-TnT (myocardial specificity protein myocardial troponin) was detected by immunofluorescent chemical method, and using real-time PCR to check c-TnT and Gata-4 gene level changes. DAPI labeled subpopulations were myocardial transplanted, and then the expression of c-TnT were detected by immunofluorescence of transplanted cells. Results CD73+ subset were mainly small cells, possessed Stellate cells, triangle cells, slender and spindle cells. In contrast, CD73- cells showed a homogeneous morphologic pattern with wide flat or square shapes, dubbed big cells. After myocardium induction, the express rate of c-TnT is 45.5% in CD73+ ADMSCs , but in the CD73- subgroup is only 5.75%. CD73+ subsets was higher than negative subgroup in expression of c-TnT and Gata-4 gene (P < 0.01). CD73+ subsets was higher than negative subgroup in expression of c-TnT in vivo also. Conclusions The percentage of cardiomyocyte transdifferentiation in CD73+ ADMSCs was obviously higher than that in CD73- subset. The results could be demonstrated that CD73+ cell possessed preferable therapy effect for heart disease.

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Division types of cultured cardiac fibroblast neonatal rat

2016, 47 (1): 55-61.

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Objective To explore the morphological features and the division mode of rat cardiac fibroblasts cultured in vitro and to provide the cellular bases for further research of cardiac regeneration mechanism and myocardial damage therapy. Methods Primary neonatal rat CFs were cultured and double labeled by vimentin and H3P, vimentin and Tubulin respectively using immunofluorescence technique. Then mitosis CFs were searched with living cell station, scanning electron microscope and a fluorescence microscope. Results Primary cultured CFs were stick wall and extended pseudopodia within 0.5-1h, keeping a great growing state with varied shapes and were rich in binucleated cells. These cells became fecund after 3 - 4 days and it is easy to find a large number of cells which were swelled up and round into mitosis phase. A few CFs in mitosis still maintained flat,and amitosis was observed incidentally. The mitosis phase of CFs in round and swelled shape was lasted about 20-30min; while for those flat ones, it was about 40-50min.In the prophase of the mitosis, the nucleus was in regular shape, and H3P began to express; then the karyotheca broke and spindle was formed in prometaphase. At metaphase, chromosomes were found in the equatorial plane, in which H3P was expressed actively. In this phase, spindles were typically in the shape of spindle. The chromatids were separated then moved towards the cellular poles; and H3P split with their separation in the anaphase. In the telophase, two daughter nuclei were formed and between them, microtubules were distributed. At cytokinesis period, two daughter cells were generated and H3P dissolved. Conclusion CFs cultured in vitro have two different morphological features – flat, round and swelled, and two division modes- amitosis and mitosis. And it is believed that these two division modes could be complementary.

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Differentiation of human bone marrow mesenchymal stem cells into corneal epithelial-like cells

2016, 47 (1): 62-66.

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Objective To investigate whether human bone marrow mesenchymal stem cells（HMSCs） are able to differentiate, in vitro, into corneal epithelial-like cells. Methods The primary human corneal keratocytes (HCFs) were obtained from the corneal stromal pieces by tissue culture, and the HMSCs were isolated by density gradient centrifugalization. Both the HMSCs and HCFs were identified according to their morphological characterizes, and the protein expression of CD29 and vimentin were analyzed by immunocytochemistry. To induce cells differentiation, HMSCs were cultured by establishing epithelial culture microenvironment in a co-culture model. Cells were characterized by phase contrast microscopy observation. Using immunocytochemistry，we confirmed the protein presence of cornea-specific cytokeratin 12 (CK12) in differentiated cells. What is more, the CK12 mRNA in differentiated cells was analyzed by reverse transcriptasepolymerase chain reaction（RT-PCR）.Results Most of the adherent HCFs and HMSCs were in spindle-shape, and perfectly align in a certain direction of the whirlpool-like shape while cells were grown to near confluence. HMSCs positively stained for CD29 and HCFs expressed Vimentin. HMSCs changing shape toward irregular polygon morphology after co-culture with HCFs over time, and the differentiating cells formed cell clusters some of which resembled epithelial-like cells. The expression of CK12 was positive for immunocytochemistry staining in differentiated cells, and further confirmed by RT-PCR. These results indicated that the differentiated cells acquired characteristics similar to those of corneal epithelial. Conclusion HMSCs isolated from human bone marrow were capable of differentiating into epithe1ial-like cells and possess the phenotypical characteristics of corneal epithe1ial.

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Gene expression analysis of hepatitis C virus related hepatocellular carcinoma genes

2016, 47 (1): 67-71.

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Objective To analyze the differentially expressed gene profiling during the development from chronic hepatitis C to HCC. Method The research uses the gene expression profile data GDS4880 and GDS4887 of the GEO database as the analysis material to make screening of the differential expression genes of the chronic hepatitis C and the HCV related HCC’s microarray data by using Qlucore Omics Explorer software, and analyze the differential expression genes and their interaction relations by using the bioinformatics tools PANTHER, DAVID , STRING and Cytoscape. Results 328 differentially-expressed genes were screened out(133 with upward effects and 195 with downward effects). The relevant genes were mainly involved in biological pathways such as metabolic process,biological regulation,localization,p53 signaling pathway, phosphatidylinositol signaling system, insulin signaling pathway, apoptosis, etc. The interaction between encoded proteins of the differential expression gene mainly focuses on the 14 proteins（CYP2B6、CYP2E1、AKT1、CDK16、RELA、CDC27、PIK3CA、GNB1、FOXO1、FYN、PDPK1、SCAND1、SGOL1、RPH3AL）Conclusion The 14 genes may be associated with the development of HCV-related hepatocellular carcinoma.

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Visualization of thalamostriate vein and its tributaries on susceptibility-weighted imaging

2016, 47 (1): 72-79.

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Objective: Thalamostriate vein is an important tributary of the internal cerebral vein which mainly drains the areas of basal ganglia and deep medulla. Susceptibility-weighted imaging is a very practical technique which is sensitive to veins, especially tiny veins. The purpose of this study was to explore the anatomic morphology of thalamostriate vein and its smaller tributaries by using SWI at 3.0Tesla and then provide the basis for the diagnosis and treatment of cerebral venous disease and the minimally invasive neurosurgery. METHODS: We acquired SWI images in 40 volunteers on a 3.0T MR system by using an 8-channel high-resolution brain phased array coil. The SWI images were reformatted by using minimum intensity projections (mIPs) technique in the transverse, sagittal and coronal plane. The anatomic morphology of thalamostriate vein and its tributaries were evaluated on the original images and reconstructed images. We classified thalamostriate vein into 2 types (Type Ⅰ and Type Ⅱ) . The anterior caudate veins were classified into 2 types (Type 1 and Type 2) and 3 Types (Type A, B and C).RESULTS: The thalamostriate vein drains the areas of caudate nucleus, the internal capsule, lentiform nucleus, the external capsule, the claustrum, the extreme capsule and the white matter of the frontoparietal lobes，except thalamus. The frequencies of the thalamostriate vein, anterior caudate veins and transverse caudate veins were 92.5%、87.5%、63.8%, separately. We found Type Ⅰ and Type Ⅱ in 79.7%, 20.3% and TypeⅠon the right side was more common than the left （P <0.05）.The most common types of anterior caudate veins were Type 1 (80.4%) and Type A(68.9%). CONCLUSION: SWI could clearly demonstrate the anatomy of the thalamostriate vein and its smaller tributaries.

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The alterations of heart and cerebral vasculature in mice after chronic cold exposure

2016, 47 (1): 80-86.

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[Abstract] Objective: The study was tried to investigate the alterations of heart and cerebral vessels after chronic cold exposure in mice. In the meantime, the pathological mechanisms were discussed as well. Methods: Male mice of ages from 90 to 180 d were used, and 60 mice were divided into control group and cold exposure group in random. Cold exposure group was fed at temperature from -1℃ and 4 ℃ for 30 days to make the chronic cold exposure model. Then, heart mass index was measured. Blood routine analyses were detected using blood cell analyzer, and the serum estrogen levels were detected by chemiluminescence method. The histological structure of cardiomyocytes was visualized with hematoxylin-eosin（H. E.）staining, and the cerebral vasculature was visualized with ink perfusion assay. Results: Chronic cold exposure could induce mouse’s myocardial hypertrophy and decrease the cerebral vascular density. The number of white blood cells, lymphocytes, red blood cells and blood platelet increased, and estrogen level of blood increased as well, which contributed to the pathological alterations of cardiovascular system. Conclusion: Chronic cold exposure could induce myocardial hypertrophy and the alterations of blood cells, cerebral vasculature and estrogen. The pathological alterations of blood cells, cerebral vasculature and estrogen are involved in the production of myocardial hypertrophy.

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Effects of Pin1 antagonist on mouse preimplantation embryo development in vitro

2016, 47 (1): 87-94.

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Objective To elucidate the further function of Pin1 in mouse preimplantation embryos, We observed the distribution of Pin1 in mouse preimplantation embryos and the effect of Pin1 antagonist (Juglone) on the development of mouse preimplantation embryo in vitro. Methods The immunofluorescence technique was used in the present study in order to localize the distribution of pin1 protein in the preimplanration mouse embryo. The 1-cell embryos of KM mice were cultured in KSOM culture medium supplemented various concentrations pin1 inhibitor (Juglone), respectively. The different development potential was observed under an inverted microscope. The mRNA expression levels of Sox2, Oct4, Klf4 and c-Myc were detected by Real-time PCR. Results Pin1 protein was distributed in both cytoplasm and nucleus of mouse preimplantation embryos. The immunofluorescent intensities in nuclei were higher than those in the complasma. The 1-cell embryos treated with 10μmol/L and 25μmol/L Juglone for 93h (from post hCG 27h to 120h) in vitro could hardly develop to 4-cell embryos (P<0.001). The 1-cell embryos treated with 25μmol/L Juglone for 18h (from post hCG 27h to 45h) in vitro were arrested at 2-cell stage (P<0.001). In 25μmol/L Juglone group, the Sox2 mRNA levels were significantly low compared with KSOM group (P<0.05), while the mRNA levels of Oct4, c-Myc and klf4 were no significant differences compared with KSOM group (P>0.05). Conclusion Pin1 is mainly distributed in the nucleus of mouse preimplantation embryos, the results suggest Pin1 can participate in gene transcription. The development rate of mouse preimplantatiom embryos from 2-cell to 4-cell is markedly decreased and the mRNA expression levels of stem cell factor (Sox2) are down-regulated by Juglone treatment, which indicates Pin1 may play a important role in the early development of mouse preimplantation embryos by regulating zygotic genome activation.

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Expression of Bmi-1 in different developmental stage of esophageal epithelium tissues

2016, 47 (1): 95-101.

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Objective To observe the Bmi-1 protein expression in esophageal epithelial tissues ,and to explore the relationship between the expression pattern of Bmi-1 and the development of esophageal epithelial tissues. Methods Immunohistochemistry was used to measure the expression of Bmi-1 in esophageal epithelial tissues of 120 cases mouse in different development time,15 cases human embryos, 18 cases normal adult human and 17 cases human esophageal squamous cell carcinoma (ESCC), then the level of Bmi-1 protein expression in these samples were analyzed by image analysis software, and the statistical analyses were applied to test the relationship between the expression of Bmi-1 and each group. Results Bmi-1 was found to expression in every time of development, but the differences were present in the protein expression level (P＜0.05). Expression of Bmi-1 protein was higher at embryo stage than that after birth both in mice and human. Compared with esophageal tissue of normal adult human, Bmi-1 overexpressed both in human esophageal squamous cell carcinoma and human embryo(P＜0.01). Conclusions The expression of Bmi-1 closely was related with the development of esophageal epithelium.Furthermore,there were a potential significance to detection of the Bmi-1 expression for the research in the developmental biology of esophageal epithelium and the cytobiology of esophageal cancer.

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Effect of different doses of salvia miltiorrhiza injection on the early structure of ovarian transplantation in mice

2016, 47 (1): 102-106.

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Objective To investigate the different doses Salvia miltiorrhiza promote follicular development in frozen-thawed mouse ovarian in an early stage after transplantation. Methods Ovaries from 1 day old mouse were cryopreserved and the thawed ovaries were xenografted into kidney capsules of 8~12 weeks old adult male mouse. The mouse were divided into four groups: Saline group、low doses(0.4ml/kg) Salvia miltiorrhiza group、middle doses (4ml/kg)Salvia miltiorrhiza group and high doses (40ml/kg) Salvia miltiorrhiza group. The grafted ovaries were collected 2, 7, and 14 days after transplantation for evaluating. Observed the histological structure and the ultrastructural changes、counted follicle numbers of the mouse ovaries after frozen-thawed and transplantation. Results The histological structure and ultrastructure showed that both the follicles and stromata were damaged on day 7 after transplantation. The follicular structure have been recovered for 14 days. The number of follicles and survival rates in grafts after transplantation of high doses Salvia miltiorrhiza group(36.2%) were more than that of middle doses、low dose Salvia miltiorrhiza group and Saline group（（24.2%、20.5、12.6%）on day 14（P＜0.05）. Conclusions Both the histological observation and ultrastructure observation methods were used in present study analyze the injury of the mouse ovarian tissues at the early stage is due to the ischemia and hypoxia rather than the cryopreservation. High doses Salvia miltiorrhiza may provide benefits for the survival rates of follicles in the frozen-thawed mouse ovarian.

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Protective effects of gastrodin pretreatment on rat isolated myocardial/ ischemia reperfusion injury

Yao Li Fei Wen Hua He Yang 无Ping Ting Ting Lai LU Di sun lin-

2016, 47 (1): 107-112.

Abstract ( )

Objective： To characterize the effect of Gastrodin on the ischemia/ reperfusion I/R injury in rat hearts. Methods： Male Sprague-Dawley rat(210-260g)were randomly assigned to 6 groups:control group(Con),ischemia/reperfusion group (I/R),pre group,pre+gas（10、50、100、200μmol/L）, The isolated hearts were reperfused in the Langendorff model and were subjected to either continuous perfusion for 90min or 30min for balance，global ischemia and reperfusion respectively. Use of cardiac electrophysiology index, myocardial enzymes and myocardial morphological changes (TTC staining) to observe the three levels gastrodin preconditioning on myocardial ischemia - reperfusion injury model, To evaluate different concentrations gastrodin myocardial ischemia - reperfusion injury in the presence or absence of myocardial protection. Conclusion： Gastrodin could protect myocardium against I/R injury in rats , which might be related with its effects of restoring cardiac function.

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Nonmuscle myosin II-B regulates the development of pharyngeal arches in zebrafish

hongkui wang xueqian wang xin wang wei yuan Dong Liu

2016, 47 (1): 113-116.

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Objective To analyze the expression of Nonmuscle Myosin II (NM II) in pharyngeal arches and the function of NM II on the development of pharyngeal arches in zebrafish embryos. Methods The expression of NM II-B in pharyngeal arches in zebrafish embryos was observed by whole-mount in situ hybridization and cryostat section (n=10). The function of NM II-B on the development of pharyngeal arches in zebrafish embryos was analyzed following the comparison of wild type (n=10) and Blebbistatin-treated (n=10 per concentration) zebrafish embryos by cartilage Alcian blue staining. Results The expression of NM II-B in pharyngeal arches in zebrafish was detected at 72 hours post-fertilization (hpf); the expression of NM II-B in pharyngeal arches increased at 120 hpf. The pharyngeal arches in zebrafish appeared obvious developmental defects, and a few of chondrocytes showed significant morphological changes after Blebbistatin inhibition of NM II-B functions, which was at a dose-dependent manner. Conclusion NM II-B regulates the development of pharyngeal arches through affecting the chondrocytes of pharyngeal arches in zebrafish embryos.

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Changes of dendritic cells in endometrial tissue of endometriosis

2016, 47 (1): 117-121.

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Objective　To investigate the expression of CD1a and CD83 which were themarkers of immature dendritic cells (imDCs) and mature dendritic cells (mDCs) respectively in endometrial tissue of endometriosis, and explore the role of DCs at different stages in the occurrence and development of endometriosis.　Methods　30 cases with chocolate cyst in the proliferative phase confirmed by pathological examination were selected, the ectopic endometrium tissues and the ectopic endometrium tissues were taken as eutopic endometrium of EMS group, ectopic endometrium of EMS group EMS and in the endometrium, another 30 cases of normal endometrial tissue were taken as control group. Immunohistochemical straining were used to detect the location and the expressions of CD1a and CD83, and western blotting were used to detect the expressions of CD1a and CD83 　Results　The CD1a and CD83 positive cells are scattered among stromal cells; The expression of CD1a increased in control group, ectopic endometrium group and eutopic endometrium group, and the difference between groups was significant (P<0.01); the expression of CD83 decreased in control group, ectopic endometrium group and eutopic endometrium group, and the difference between groups was significant (P<0.01). Conclusion Compared with non EMS patients, the increase of imDCs and the decrease of mDCs in local endometrium in endometriosis may play important roles in the occurrence and development of EMS.

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Fasciology

2016, 47 (1): 121-121.

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Correlation between muscle mass and fat mass and limb circumference basis on electrical impedance in adults of He’nan Hui ethnicity

xu guochang Chang-De MA Nuan LIU XU Fei Yong LIU

2016, 47 (1): 122-128.

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Regression analysis of male maxilla and skull

2016, 47 (1): 129-133.

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Objective:To study and establish the regression equation applied to estimate the main general skull diameter index with the maxilla index. Methods: Totally 135 adult male mandibles were used for 37 items of measurements with the physical anthropology measuring method,the data were studied by statistical software (SPSS 17.0).Eliminating the r<0.4 and poor correlation pairing. Results:Totally 78 ong-factor line regression equations were set up,which the P vaoues were than 0.001.Conclusion: These equations can be used to estimate the main general skull diameter index with maxilla index, and to provide evidences for the recovery of the skull.

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Analysis of the body composition of adult Tibetans in Gansu and Tibet

2016, 47 (1): 134-138.

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[Abstract] Objective Exploring the body composition of Chinese Tibetans by collecting and comparing the body composition data of adult Tibetans in Gansu and Tibet of China to provide basis for further researches on the relationship between body composition and living surrounding as well as the incidence of chronic diseases. Methods Bioelectric impedance technique was used to test the body composition indices of cluster sample of 814 adult Tibetans living in Shigatse, Tianzhu Tibetan Autonomous County and Gannan Tibetan Autonomous Prefecture. Results The BMI, lean body weight, body fat rate, subcutaneous fat , visceral fat, muscle amount, fluid amount, protein amount and WHR in male Tibetans reached their maximum at their40~50 years old; the basal metabolism decrease with the increase of their age after 40~50 while their extracellular fluid to intracellular Fluid ratio(E/I) raise with the aging; as to female, their lean body weight, muscle amount and basal metabolism reached the peaks at their 30~40 years old while the fluid amount maximum is at their 40~50 years old; BMI, body fat rate, subcutaneous fat , visceral fat, WHR and E/I increase with aging whereas the protein decrease with the increase of their age. Compare with the participants in Tibet, the Tibetans from Gansu have higher BMI, lean body weight, muscle amount, subcutaneous fat , visceral fat, body fluid amount and basal metabolism as well as more overweight , obesity; however, the prevalence of high WHR in Tibetans in Tibet is higher than the ones from Gansu (p＜0.05). Conclusion The variation of body fat indices in Tibetan males and females of are sinusoid and increasing with aging, Tibetans from different regions exhibit different body composition values; most of body composition indices especially fat related indices of Tibetans from Gansu are higher than participants from Tibet, which may lead to higher incidence of chronic disease in Gansu Tibetans.

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Established the 3D anatomy interaction system of zebrafish (Danio rerio)

2016, 47 (1): 139-144.

Abstract ( )

Objective The aim of this paper was to make a software of three-dimension (3D) anatomical interaction system in zebrafish (Danio rerio). Methods The structure of zebrafish was observed by light microscopy and Micro-CT, and the images of its morphological structure were collected. The pictures of serial section (HE-stained) were corrected by Photoshop. The 3D model was established by the software of Autodesk 3ds Max. Results The results showed the zebrafish skeletal 3D imaging and serial section images (male: 144, female: 179). We realized the 3D reconstruction of the vital organs of zebrafish, and established the 3D anatomy interaction system zebrafish (V 1.0), which obtained the computer software copyright registration certificate (Certificate Number: soft word board the Di_0989966). Conclusion The zebrafish 3D anatomy interaction system software afforded the three-dimensional morphological basis for the study of zebrafish, and was helpful to histomorphology recognization for researchers.

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