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Table of Content

    2016, Volume 47 Issue 3
    06 June 2016
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    Effects of ischemia and hypoxia on DNA methylation in mouse hippocampus
    LI Hong SHI Zhen-yu LI Rui-ling ZHANG Pei FU Su DENG Jin-bo* DENG Jie-xin*
    2016, (3):  289-296.  doi: 10.16098/j.issn.0529-1356.2016.03.001
    Abstract ( )  

    Objective To establish a hypoxic-ischemic brain slice model, and to investigate the neuronal injuries and DNA methylations on the mouse hippocampus after hypoxic-ischemic brain damage. Methods Healthy adult C57BL / 6J mouse hippocampal slices were used to establish a HIBD model. The hypoxic-ischemic model was tested with TTC staining, and the changes of inflammatory injury and the expression of enzymes relating to DNA methylationwas were visualized by immunohistochemical and Western blotting. Results Cells’ oxidative stress and inflammatory damage of HIBD hippocampal slices were much more serious than that in the control group (P<0.01), while the level of DNA methylation was higher, compared to the control group (P<0.01). Conclusion Hypoxic-ischemic can induce the hippocampal inflammatory damages, and enhance the level of DNA methylation, suggesting that DNA methylation is involved in the process of hypoxic-ischemic tissue injury. The relative mechanism may be that HIBD can cause the DNA methylation. Conversely, DNA methylation can increase oxidative stress and hippocampal damage.

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    P2X3 mRNA expression in L4-6 dorsal root ganglion at different times after the spared nerve injury
    DU Jun-ying FANG Jun-fan FANG Jian-qiao* LE Xiao-qin XIAO Ting PAN Ning-fang YU Jie
    2016, (3):  297-302.  doi: 10.16098/j.issn.0529-1356.2016.03.002
    Abstract ( )  

    Objective To observe the mRNA expression of P2X3 in L4-6 dorsal root ganglion (DRG) at different times after spared nerve injury (SNI), and to discuss the effect of periphery P2X3 in neuropathic pain. Methods Fifty-four healthy male SD rats were randomly divided into control group, sham surgery group, and surgery group. The spared nerve injury model was established by ligating the common peroneal and the tibial nerves and then cutting off the nerves but keeping the sural nerve intact. Bilateral paw withdrawal thresholds were dynamic observed before injury (base), at day 1, 3, 7 and day 14 after injury. P2X3 mRNA expression in L4-6 DRG at day 3, 7, and day 14 after injury was measured by fluorescence quantitative PCR method. Results Compared with control group, ispilateral paw withdrawal thresholds of surgery group were significantly decreased at day 1, 3, 7, and day 14 after injury (P<0.05), while that of sham surgery had no statistical significance (P>0.05). There was no difference of contralateral pain threshold among control group, surgery group, and sham surgery group (P>0.05). P2X3 mRNA expressions of L4, L5, and L6 DRG of surgery group were remarkable increased at day 3, 7 after injury (P<0.05), while that of L5, L6 DRG were decreased at 14d after injury (P<0.05), and that of L4 DRG were more than that of sham surgery at day 14 after injury (P<0.05). In addition, there was no difference of P2X3 mRNA expression in L4, L5, and L6 DRG at every time after injury between sham surgery group and surgery group. Conclusion Periphery P2X3 mRNA expression is involved in the initiation and maintenance of pain induced by SNI, which may play a different effect at different phases.

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    An improved glial scar model after central nervous system injury in vitro
    LI Yi TAN Ling LIN Wei-wei CHEN Xue CHEN Ying PAN Jing-ying WANG Xiao-dong*
    2016, (3):  303-308.  doi: 10.16098/j.issn.0529-1356.2016.03.003
    Abstract ( )  

    Objective To establish a scar model in vitro and observe the morphology and extracellular matrix of the scar. Methods Astrocytes and meningeal fibroblasts were cultured from the cerebral cortex in vitroand identified by glial fibrillary acidic protein (GFAP) and fibronectin(FN) immunofluorescence cell staining respectively. Astrocytes and meningeal fibroblasts were co-cultured and treated by TGF-β1 after 2 days. The group without TGF-β1 treating was used as the control. The morphology of scar was observed by GFAP and FN immunofluorescence cell staining. Immunofluorescence cell staining and Western blotting were used to examine the expression of EphB2, ephrinB2, neurocan and NG2 in the scar. Results The scar-like cell cluster was formed from astrocytes and fibroblasts. In the TGF-β1 group, the expressions of EphB2 and ephrinB2, neurocan and NG2 were significantly increased when compared with the control group (P<0.05). Conclusion A scar model after central nervous system injury has bean established by co-culturing astrocytes and fibroblasts treate by TGF-β1 in vitro.

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    Quality analysis of 100 tissue samples from the postmortem human brain bank
    QIU Wen-ying LIU Fan YANG Qian LIU Wei WANG Nai-li ZHANG Di WANG Tao YUAN Bo MA Chao*
    2016, (3):  309-314.  doi: 10.16098/j.issn.0529-1356.2016.03.004
    Abstract ( )  

    Objective To perform quality analysis for the brain tissue samples from the postmortem Human Brain Bank in Chinese Academy of Medical Sciences and Peking Union Medical College. Methods DNA, RNA and protein were extracted from a total of 100 deep-frozen tissue samples stored in the brain bank, followed by absorption photometry, electrophoresis,Western blotting, RT-PCR and Real-time PCR analysis. The samples were grouped according to age, cause of death, post-mortem delay time and duration of storage. Results The concentrations of DNA, RNA and protein and the integrity of DNA and RNA (accessed via A 260/280) were compared among groups. The results indicated that there were no major DNA, RNA and protein degradation in the brain tissue with postmortem delay less than 24 hours, and the concentrations were not affected by age and cause of death. In the samples with post-mortem delay over 24 hours, the concentration of protein and RNA as well as the RNA integrity declined significantly. Protein and DNA integrity were not affected by the time of storage, while RNA concentration decreased when the storage time was over 1 year. Conclusion The postmortem delay should be controlled within 8 hours for RNA analysis, but may be up to 24 hours for protein analysis and even longer for DNA. The human brain tissue can be preserved below -80℃ for as long as 1 year without affecting the sample quality.

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    Expression changes of AKT, ATM, D4-GDI and p53 cell apoptosis pathway genes in the process of rat liver regeneration
    XING Xue-kun LI Meng-hua XU Cun-shuan *
    2016, (3):  315-322.  doi: 10.16098/j.issn.0529-1356.2016.03.005
    Abstract ( )  

    Objective To investigate the expression changes of four cell apoptosis pathway genes, AKT, ATM, D4-GDI and p53 in the process of liver regeneration.
    Methods The rats were randomly divided into enperiment group and control group, 6 rats in each group, half male and half female. Genome Rat 230 2.0 was used to detect the expression of genes in four cell apoptosis pathways in rats after partial hepatectomy (PH), and the biological information method was used to analyze it. Results 63, 8, 6 and 7 genes were associated with liver regeneration in the four cell apoptosis pathways.They were mainly expressed at the start stage of liver regeneration. The similarity of their expression was including totally up regulation, up regulation advantage, totally down regulation, down regulation advantage, up regulation equal to down regulation of 5 categories. The expression of most genes in liver regeneration was enhanced, and the expression of few genes was decreased. Time correlation was divided into 11 groups. Analysis of the role of apoptosis pathway related genes in liver regeneration by using the gene synergy model (Et) indicated that AKT pathway inhibited the apoptosis of cells in the whole liver regeneration; ATM, D4-GDI and p53 three cell apoptosis pathway in the promotion of cell apoptosis in the whole liver regeneration. Conclusion Four apoptosis pathways, AKT, ATM, D4-GDI and p53, are closely related to the growth, development, and liver volume control of liver regeneration.

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    Regulatory role of GADD45α in rat liver regeneration
    YANG Xian-guang ZHU Lin ZHAO Wei-ming HE Chun-cui XU Cun-shuan*
    2016, (3):  323-329.  doi: 10.16098/j.issn.0529-1356.2016.03.006
    Abstract ( )  

    Objective To explore the role and mechanism of growth arrest and DNA damage inducible gene GADD45α during liver regeneration (LR) after partial hepatectomy (PH) in rats. Methods 114 rats were randomly divided into 19 groups, 9 groups of 2/3 liver resection, 9 control groups and 1 normal control group. The rat PH model was established, rat Genome 230 2.0 array was used to detect the gene expression profiles of the regenerated liver after PH, and Real-time PCR technique was used to verify the reliability of the microarray. Expression profile function (Ep) was used to analyze the biological processes that regulated by GADD45α, and then Ingenuity Pathway Analysis (IPA) was used to summarize the signal pathway of GADD45α regulating LR and analyze its possible mechanism. Results The microarray results showed that Gadd45α was up-regulated at 2-6 hours, 24 hours and 36-72 hours after PH. The IPA results indicated that GADD45α regulated various physiological activities which were involved in rat LR via NF-κB, p38PRAK, p53, STAT3-p21, STAT3-Bcl-2 and STAT3-cMyc paths. The results revealed that changes of physiological activity were in line with the basic process of rat LR. Conclusion GADD45α may regulate cell proliferation, cell cycle and cell survival and other physiological activities during the regeneration of liver in rat through the above pathways.

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    Protective effects of ethyl acetate extract from Chroogomphis rutilus on SH-SY5Y cells damaged by 1-methyl-4-phenyl pyridine ion
    LI Ge YAO Hui LIU Chuan GAO Zhi-guo WANG Li-an GUAN Zhen-long WANG Yan-qin*
    2016, (3):  330-336.  doi: 10.16098/j.issn.0529-1356.2016.03.007
    Abstract ( )  

    Objective The aim of present study is to observe the protective effects of ethyl acetate extract fromChroogomphis rutiluson dopaminergic neurons damaged by 1-methyl-4-phenyl pyridine ion (MPP+). Methods SH-SY5Y cells (dopaminergic cells), a cell line of human neuroblastoma, were served as the experimental subject. We examined the IC50 of MPP+ at 48 hours. The cells were preincubated with ethyl acetate extract fromChroogomphis rutilusfor 4 hours with 25mg/L, 50mg/L, 100mg/L respectively, then added MPP+ 1.0mmol/L(IC50 value) to incubate for 48 hours. MTT assay, Hoechst 33342 staining and flow cytometric assay were used to detect the survival or apoptosis of SH-SY5Y cells. DCFH-DA was used to detect the reactive oxygen speciess (ROS) level on the cells of ethyl acetate extract fromChroogomphis rutilus. Results 1.0mmol/L MPP+ caused the IC50 of cells at 48 hour, so 1.0mmol/L was used as the lesion concentration. Pretreatment with ethyl acetate extract fromChroogomphis rutilus (50mg/L) significantly increased the cell viability, and reduced the apoptosis rate of cells. After MPP+ lesion,the ROS level increased significantly, and the preincubation with ethyl acetate extract fromChroogomphis rutilus reduced the ROS level induced by MPP+. Conclusion Ethyl acetate extract from Chroogomphis rutilus can significantly protect dopaminergic cells damaged with MPP+.

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    Expression and significance of synaptopodin and cytochrome C in podocytes injuries induced by palmitate acid
    QI Fang ZANG Dong-yu AI Hao LI Xiao-ming*
    2016, (3):  337-340.  doi: 10.16098/j.issn.0529-1356.2016.03.008
    Abstract ( )  

    Objective To investigate the expression and significance of synaptopodin and cytochrome C(Cyt C) in podocyte injuries induced by palmitate acid. Methods Podocytes were cultured and randomly assigned into two groups, normal group and palimitate acid group.The two groups were cultured for 24hours, 72hours and 120hours. Cell viability of podocytes induced by palmitate acid was examined by MTT. The expressions of synaptopodin and Cyt C of podocytes were examined by immunofluorescence and western blotting. Results Compared with the normal group, cell viability of podocytes was decreased in the palimitate acid group in a time dependence manner. The immunofluorescence and Western blotting results showed that expression of Cyt C was enchanced in podocytes induced by palmitate acid. The expression of synaptopodin was reduced in podocytes induced by palmitate acid. Conclusion Palmitate acid induces that Cyt C is released and synaptopodin is abnormally expressed in podocytes, which may result in podocyte injuries eventually.

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    Effects and mechanism of olanzapine on the adipose differentiation of mouse bone marrow mesenchymal stem cells
    JI Chen-yanLI Yong-hai*LI MengYANG Hai-jie FENG Zhi-wei
    2016, (3):  341-347.  doi: 10.16098/j.issn.0529-1356.2016.03.009
    Abstract ( )  

    Objective To investigate the effects of olanzapine on the adipose differentiation and mechanism of mouse bone marrow mesenchymal stem cells(BMSCs).
    Methods MTT colorimetry was used for assaying different concentrations of olanzapine on the proliferation of mouse BMSCs. The morphology of cells was observed by Oil red O staining. The expressions levels of adipocyte markers αP2 and C/EBPβ were detected by Western blotting, mRNA expression levels of the related genes of Leptin, C/EBPα and TNF-α were analyzed by Real-time PCR. The protein expression of Akt signaling pathway related molecules and effects of phosphatidylinositol 3 kinase (PI3K)/Akt inhibitor on the differentiation of BMSCs were determined by Western blotting. Results The results of MTT colorimetry showed that 20μmol/L olanzapine had minimal toxicity on BMSCs. Oil red O staining showed that intracellular lipid droplets in the experimental group were significantly more than the control group. The results of Western blotting showed that the expression levels of αP2 and C/EBPβ increased by about 36%(P<0.01) and 25%(P<0.05) compared with the control group respectively. The results of Real-time PCR showed that the expression levels of adipogenic genes Leptin, C/EBPα and TNF-α significantly increased than the control group, increased about 68%(P<0.001),79%(P<0.01)and 60%(P<0.01)respectively. The expression levels of p-Akt were significantly higher than the control group, the expression of glycogen synthase kinase 3β(GSK-3β) with the increase of p-Akt expression gradually reduced. The expression levels of αP2 and C/EBPβ were inhibited after BMSCs were pretreated with PI3K/Akt inhibitor (LY294002). Oil red O staining results showed fat droplets in cells also decreased significantly. Conclusion Olanzapine may elevate the expression level of p-Akt of PI3K/AKT signaling pathway to promote the adipogenic differentiation of mouse BMSCs.

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    Bioinformatics analysis of the genes related to the pregnancy associated breast cancer
    TENG Mu-zhou CHEN Li-na LU Yan-fang MA Wen-li*
    2016, (3):  348-352.  doi: 10.16098/j.issn.0529-1356.2016.03.010
    Abstract ( )  

    Objective The differentially expressed genes of pregnancy associated breast cancer patients and normal subjects were analyzed by Bioinformatics to reveal the pathogenesis of pregnancy associated breast cancer on the molecules level and to provide new ideas for the further study on breast cancer. Methods The microarray data sets of pregnancy associated breast cancer were downloaded from the public gene expression database (Gene Expression Omnibus, GEO); differential genes of pregnancy associated breast cancer patients and normal subjects were selected by Qlucore Omics Explore (QOE); DAIVID, and STRING were adopted to analyze the function and signal pathway and to predict the protein-protein interaction of the differential genes. Results A total of 148 differentially expressed genes were screened, among which 24 were up-regulated and the other 124 were downregulated. The results of these 148 differential genes bioinformatics analysis showed that the genes TAGLN, ACTG2, TPM2, TPM3, MYLK, ACTA2, MTH11, and mitogen activated protein kinase(MAPK) signaling pathway, and focal adhesion pathway, and vascular smooth muscle contraction pathway may play important roles in the development of pregnancy associated breast cancer. The results of STRING analysis showed that 20 genes were located in the key nodes of the protein interaction network. Conclusion Bioinformatics method can be utilized to analyze microarray data effectively andmining the deeper information of the data, providing valuable clues for the further researches.

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    Sodium selenite induces apoptosis of human gastric cancer SGC-7901 cells through mitochondrial-dependent pathway
    WANG Yan ZHAO Shang SU Yan-ping* LIU Li-wei WANG Hui QU Peng*
    2016, (3):  353-358.  doi: 10.16098/j.issn.0529-1356.2016.03.011
    Abstract ( )  

    Objective To study the mechanism of gastric cancer cells apoptosis induced by sodium selenite,by investigating the expression of Bax/Bcl-2, the change of mitochondrion membrane potential and cytochrome C(Cyt C) in the process apoptosis ofof human gastric cancer SGC-7901 cells. Methods Human gastric cancer SGC-7901 cell line was added different concentrations (2.5, 5.0, 10.0μmol/L) of sodium seleite for 24hours,48hours. The expression of Bax/Bcl-2 protein was detected by immunocytochemical and Western blotting methods. SGC-7901 cells were stained by Rhodamine123 to detect changes of mitochondrion membrane potential by flow cytometry. The changes of cytoplasm Cyt C and mitochondria Cyt C protein content were detected by Western blotting. Results Immunocytochemistry and Western blotting showed that sodium selenite increased the Bax protein levels(P<0.05) and reduced the Bcl-2 protein levels (P<0.05) in dose-dependent manner.Flow cytometry technique results showed that sodium selenite reduced mitochondrial membrane potential(P<0.05). The results indicated that sodium selenite may induce SGC-7901 cells apoptosis by altering Bax/Bcl-2 and reducing mitochondrial membrane potential. Western blotting showed that sodium selenite increased the cytoplasm Cyt C protein levels(P<0.05) and reduced the mitochondria Cyt C protein levels (P<0.05), therefore promoted the release of Cyt C to cytoplasm. Conclusion Sodium selenite can induce apoptosis of human gastric cancer SGC-7901 cells by increasing Bax and discreasing Bcl-2 gene expression, reduce the mitochondrial membrane potential, to promote Cyt C release from mitochondria to the cytoplasm, and induce human gastric cancer SGC-7901 cell apoptosis through mitochondrial-dependent pathway.

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    Comparison between venous superdrainage and arterial supercharging on flap survival
    ZHENG Jun XI Shan-shan LI Hong DING Mao-chao HAO Xiao-di MEI Jin TANG Mao-lin CHEN Shi-xin*
    2016, (3):  359-364.  doi: 10.16098/j.issn.0529-1356.2016.03.012
    Abstract ( )  

    Objective To compare the effects of venous superdrainage and artery supercharging to perforator flap hemodynamics, therefore, reducing the rate of flap necrosis, improving their survival area, and providing some basis for flap survival. Methods Sixty Sprague-Dawley rats, weighing (500±50)g, were divided into three groups: experimental group A (Exp A), experimental group B (Exp B), and control group (control)), 20 rats each group. In the experimental group A, the posterior intercostal artery was ligated while the accompanying vein was reserved. In the experimental group B, the posterior intercostal vein was ligated while accompanying artery was reserved. In the control group, the posterior intercostal artery and vein were ligated. Laser Doppler imaging was used to evaluate flap perfusion in the two “choke area”, 6 h, 1day, 2days, 3days and 7days after surgery. On postoperative day (POD) 7, survival areas of flaps were calculated and lead oxide-gelatine was used for vascular angiography. Results In Exp A, all flaps survived (98.2±1.6)%; in Exp B, flap survival area was (74.78±5.91)%; in Control, survival area was (60.3±7.8%)(P<0.001). From 6 hours to 7days postoperatively, blood flow or oxygen partial pressure of flaps in Exp A was significantly higher than Exp B and Control(P<0.05). On angiography, anastomosisof the choke vessels in Exp A was richer than in Exp. B and Control. In Control, flap necrosis due to poor blood circulation, vascular anastomosis and distal venosome were not seen. Conclusion The effect of venous superdrainage was more apparent than arterial supercharging, and may effectively alter hemodynamics of microcirculation, and improve survival area of skin flap.

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    Applied anatomy of far lateral suboccipital approach and its clinical experience
    JI Yun-xiang WANG Ye-zhong* ZHAO Dong ZHU Li-cang DAI Lin-zhi TIAN Wei-dong
    2016, (3):  365-367.  doi: 10.16098/j.issn.0529-1356.2016.03.013
    Abstract ( )  

    Objective To analyse the operative results of removing craniocervical junction region (CCJR),and its surgical techniques. Methods Micro-anatomy of craniocervical junction region was studied in 10 adult cadaver heads for simulation of far lateral suboccipital approach. The structure of the extension neck junction was observed. The far lateral suboccipital opproach was used on 26 cases with lesions surgery of the anterior or anterolateral CCJR. Results Of 26 patients, 9 patients were recovered and 17 patients improved. There were 14 cases of tumor resection, and 12 cases of subtotal resection. There was no operative mortality. Postopertive complications included facial paralysis in 2 cases, mild cranial nerve palsy in 1 case, and cerebrospinal fluid leakage in 1 case. Conclusion The far lateral suboccipital approach can extensively reveal the surgical field of the anterior and anterolaeral CCJR, and is an effective surgical resection of the lesion site.

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    Effects of the ephedrine injection followed by a stomach irrigation of Chinese medicine compound in the pregnant mouse on hepatic tissue structure and antioxidant enzymes activity of the filial mice
    LI Chong-yang YANG Ya-fei ZHAO Wen-cheng YU Shi-yuan*
    2016, (3):  368-373.  doi: 10.16098/j.issn.0529-1356.2016.03.014
    Abstract ( )  

    Objective To explore effects of ephedrine injection followed by the stomach filling of Chinese medicine compound in the pregnant mouse on the development of the filial mice. Methods Totally 30 pregnant mice were used in the present study and divided into 3 groups,the natural control group,experimental control group and experimental group.The pregnant mice of experimental control group and experimental group were continuously intraperitoneally injected 6.0 g/L ephedrine (twice a day, 0.2ml each),the natural control group was injected with the same amount of saline.The pregnant mice of experimental group were irrigated to stomach with 30.0 g/L Chinese medicine compound 0.2ml 1 hour after ephedrine injection.The natural control group and experimental control group were irrigated to stomach with same amount of saline.On day 5,10 and 15 after delivery,the activities of filial mice hepatic histological total antioxidant capacity(T-AOC),glutathione S-transferase(GST)and plasma alanine aminotransferase(ALT),as well as content of malondialdehyde(MDA)MDA were determined by colorimetry,and the liver tissue structures of filial mice were observed by optical microscopy,the expression changes of TGF-β1 protein were measured by immunohistochemical staining. Results Compared with natural control group the activity of filial mice’s hepatic histological T-AOC,and GST of experimental control group were significantly reduced,while the content of MDA was increased significantly,the activity of plasma ALT was significantly increased,the filial mice’s liver tissue appeared varying degrees of damage,the expression intensity of TGF-β1 protein in liver tissue was significantly increased (P<0.05 or P<0.01).But the activity of T-AOC,GST of the filial mice’s liver tissue in the experimental group were increased significantly,the content of MDA was significantly decreased,and the activity of plasma ALT was significantly reduced.The pathological damage of liver was significantly improved,the expression of TGF-β1 protein was decreased (P<0.05 or P<0.01). Conclusion Ephedrine affects the histological structure and antioxidant enzymes activity and TGF-β1 expression of the liver.The female mice with stomach irrigation of Chinese medicine compound may enhance filial mice liver cells antioxidant enzymes activity,reduce TGF-β1 protein expression of liver tissue,and lighten the damage of filial mice liver tissue by ephedrine injecting of female mice.

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    Comparison of microstructures of the carotid body between yaks and chaidamu yellow cattles
    CHANG Lan ZHANG Shou LEI Nai-hu MA Yan-fang BAI Zhen-zhong SHEN Ming-hua GE Ri-li*
    2016, (3):  374-380.  doi: 10.16098/j.issn.0529-1356.2016.03.015
    Abstract ( )  

    Objective To compare the structural characteristics of the carotid body (CB) between yaks and chaidamu yellow cattles. Methods Health adult yaks (n=5) and chaidamu yellow cattles (n =5) were used in this study. The microstructure and ultrastructure of CB were observed by histologial, immunohistochemical SP methods and transmission electron microscopy techniques. The volume density(Vv), surface density(Sv), numerical density on area(NA) and specific surface(δ) of type Ⅰ cells(light and dark) and type Ⅱ cells and microvessels of carotid body were compared by light microscopy and microstereology between yaks and chaidamu yellow cattles. Results The CBs of yak and chaidamu yellow cattle were composed of parenchyma and mesenchyme. There were many round or oval gloubs with rich parenchymal cells. There were more type Ⅰ cells in yaks than in chaidamu yellow cattles, but there were less and smaller mitochondria and electron densecored vesicles(EDCV) of the type I cells in yaks than in chaidamu yellow cattles. The connective tissue among gloubs and interstitial cells in chaidamu yellow cattles were significantly more than yaks, but there were more rich capillaries in mesenchyme and bigger vascular lumen in yaks than in chaidamu yellow cattles. The Vv、Sv、NAof CB light cells in yaks were significantly greater than that in chaidamu yellow cattles (P<0.01),while δ of CB light cells in yaks was less than in chaidamu yellow cattles(P>0.05). The Vv、Sv of CB dark cells in yaks were significantly less than in chaidamu yellow cattles (P<0.05), and there was no significant difference of NAand δ between yaks and chaidamu yellow cattles(P>0.05). In type Ⅱ cells, there was no significant difference in Vv、Sv、NA、δ between two groups (P>0.05). The Vv、Sv、NA of the CB microvessels in yaks were obviously greater than that in chaidamu yellow cattles (P<0.01), while δ in yaks CB was significantly less than in chaidamu yellow cattle (P<0.01). Conclusion The inheret tissue cell composition and vascular net of yak CB are formed to enhance the adaptability to plateau hypoxic environment.

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    Effects of secretory diarrhea on calcium activated chloride channel gene expression in the mouse
    LI He-ping GUAN Yan-jing ZHA Guang-ming WANG Xin-jian WANG Yue-ying*
    2016, (3):  381-385.  doi: 10.16098/j.issn.0529-1356.2016.03.016
    Abstract ( )  

    Objective To investigate effects of secretory diarrhea on calcium activated chloride channel(CLCAs)gene expression in the mouse intestinal tissue. Methods Twenty-four Kunming mice (12 males and 12 females) were randomly divided into three groups (n=8 in each group): control, experimental 1 hour and 8 hours groups. The control group were given a normal saline intraperitoneal injection of 0.2 ml, the experimental group were treated an by intraperitoneal injection of lipopolysaccharide (LPS,6 mg/kg) for 1hour or 8hours respectively. After injection for 1 hour or 8 hour, the mouse mental state and intestinal morphology were observed to evaluating secretory diarrhea model establishment. Real-time PCR was used to detect CLCAs gene expression in the intestine. Results The results showed that, LPS induced secretory diarrhea in mice, CLCAs genes were expressed in several intestine segments. After LPS injection, the transcriptional levels of CLCA1, CLCA2, CLCA3, CLCA4, CLCA6 gene in duodenum, jejunum, ileum and colon were up-regulated. Compared with 1 hour group, the gene transcription level significantly up-regulated in 8 hours group. After LPS injection, CLCA5 expression was not detected in duodenum, but expression of CLCA5 was significantly up-regulated in jejunum and ileum. Conclusion The different CLCAs gene expression in intestinal segments is related to secretory diarrhea caused by LPS.

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    Effect of Guizhou liquor on rat Leydig cells in the expression of steroidogenic acute regulatory protein and serum testosterone level
    LIU Hao LIANG Wen-mei*
    2016, (3):  386-390.  doi: 10.16098/j.issn.0529-1356.2016.03.017
    Abstract ( )  

    Objective To investigate effects of a Guizhou 54 degrees liquor on rat leydig cells in the expression of steroidogenic acute regulatory protein (StAR) and the change of the serum testosterone level at the different time and with different doses. Methods Sixty normal adult male SD rats were randomly divided into a normal control group(n=15) and an experimental group(n=54). Each experimental group was further subdivided into low dose group 0.8 ml/(kg·d), middle dose group 1.6 ml/(kg·d), high dose group 2.4 ml/(kg·d). The rats of the experimental groups were given by gavage twice a day(11∶00 AM an 5∶00 PM)with liquor. At the end of the 4th, 8th and 12th week, the serum was collected and testicular tissue was taken. The expression of StAR was detected by immunohistochemical SABC method, image analysis and Western blotting, and level of the serum testosterone was detected by Chemiluminescence method.
    Results StAR immunohistochemical positive products existed within the cytoplasm of Leydig cells; Compared with the normal control group, the average absorbance of StAR staining in the experimental groups at the 4th, 8th, and 12th week increased(P<0.05). Western blotting showed that the expression level of StAR protein in the experimental groups at the 4th, 8th and 12th week increased(P<0.05). Compared with normal control group, serum testosterone level of each experimental group did not see apparently unusual,and there was no statistically significant difference(P>0.05). Conclusion In this experiment a set of doses and schedules after gavage with the liquor enhance the StAR expression level in the rat Leydig cells enhancement.Serum testosterone level has no obvious change.

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    Expression of integrin β1 and focal adhesion kinase of the left ventricule in renal hypertensive rats
    TIAN Xiang-qin LIU Ying-chun CAI Xin-hua*
    2016, (3):  391-395.  doi: 10.16098/j.issn.0529-1356.2016.03.018
    Abstract ( )  

    Objective To explore the variation of integrin β1 and focal adhesion kinase(FAK)during left ventricular wall re-modeling in the rats with renal hypertension.
    Methods The animal model was prepared by bilateral renal artery stenosis, and renal artery was freed in the control group. Forty rats were randomly divided into sham operation group, and two-week group, five-week group and seven-week group after operation. The model quality of hypertensive rat was assessed by a noninvasive blood pressure analysis tester and the index of left ventricular weight (LVW/BW). The morphological changes of the left ventricle was observed under a light microscope after HE staining. The immunohistochemistry and Western blotting methods were used to detect integrin β1 and FAK distribution in the ventricle muscle tissues. Results HE staining showed that the myocardial fibers were disordered and the space between the myocardial cells increased. The positive signals of both of integrin β1 and FAK immunohistochemistry were distributed in myocardium. The intensities of the immunohistochemical positive staining profiles for both integrin β1 and FAK increased with time and reached equilibrium for 5weeks treatment by image analysis. Average absorbance(AA) value of FAK expression at the 2nd week was declined significantly than the control group and increased with time in operation groups. Western blotting showed that the expression level of integrin β1 and KAK in every operation group were all higher than sham group (P<0.01). Five-week group was declined significantly compared with 2-week group (P<0.01) and no significance difference between 7-week and 5-week group(P>0.05).FAK had a significantly lower expression level in all operation groups than control group, 5-week group was declined significantly compared with 2-week group (P<0.01) and no significance difference between 7-week and 5-week group(P>0.05). Conclusion The abnormal expression of integrin β1 and FAK in the left ventricular may be an indication of myocardial hypertrophy.

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    Bioelectrical impedance analysis of non-fat tissue body composition in Dongxiang nationality adults of Gansu
    YE Zhen-zhen HE Ye* HAI Xiang-jun MA Wei-hong HE Jin-quan MA Bin GU Qiao-ling SU Lu YANG Tao
    2016, (3):  396-403.  doi: 10.16098/j.issn.0529-1356.2016.03.019
    Abstract ( )  

    Objective To analyze the changing characteristics of non-fat tissue body composition in Dongxiang adults of Gansu by a bioelectrical impedance analysis method. Methods A total of 491 Dongxiang adults aged over 20 years in Dongxiang autonomous county of Gansu Province were selected by the cross-sectional stratified random sampling method. The non-fat tissue body composition indexes were measured by body composition analyzer. Results The mass to constituent ratio of lean body mass, bone mass, muscle mass, protein and total body water were higher in male adults of Dongxiang in Gansu than in female (P<0.01), and bone mass, muscle mass, protein and total body water were same in different age groups. Bone mass and muscle mass showed the same trend in the same gender. One of the peaks of them was in 40 age group in male adults, and they decreased at first and then increased up to peak in 40 age group in female. The protein in male decreased after the peak of 30 age group, and decreased in female. Trunk muscle mass in male and female decreased after the peaks of 50 age group and 40 age group respectively. Double upper limbs muscle mass and total body water in both decreased after the peak of 40 age group. Double lower limbs muscle mass in male showed a downward trend except that there was a slight rebound in 40 age group, and always decreased in female. Intracellular water decreased at first and then increased up to peak in 40 age group in both. Extracellular water in male and female decreased slightly after the peaks of 50 age group and 40 age group respectively. Conclusion Non-fat tissue body composition indexes in Dongxiang adults of Gansu in male are higher than in female in different age groups and all of them changed with age. There are gender differences in the trend of the remaining body composition except double upper limbs muscle mass, total body water and Intracellular water. In general, the characteristics of non-fat tissue body composition in female adults of Dongxiang are less mass than in male, decrease continuously with no peak of partial body composition or earlier peak, decrease earlier, and decrease more slowly than in male.

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    Differences of headface morphological traits between southern and northern Chinese Han
    YU Ke-li ZHENG Lian-bin* LI Yong-lan XI Huan-jiu LU Shun-hua ZHANG Xing-hua BAO Jin-ping
    2016, (3):  404-408.  doi: 10.16098/j.issn.0529-1356.2016.03.020
    Abstract ( )  

    Objective To Compare and discuss head-face morphological traits between Southern and Northern Chinese Han. Method From 2009 to 2012, we measured northern Han head-face anthropometry indices of 11 732 cases (male 5840, female 5892) and made statistical tests of differences between the northern Han and the southern Han. Results Compared with the southern Han, northern Han head was wider and higher, bigonial breadth was wider, the distance between the two eyes was bigger, the surface was higher, nose was higher, nose and lips distance was larger, ear was big, but the head was shorter, surface was narrower, nose was shorter, nasal was narrower, and lips were thinner. Compared with the southern Han, the northern Han male head was more round, higher and narrower, surface was narrower, and nasal was more broad. The northern female was the same except nasal. Female nasal was narrower. Conclusion Genetic factors are the main factors that affect the formation of head-face.

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    Relationship between androgen receptor CAG/GGN repeat polymorphisms and the ratio of 2D∶4D
    ZHANG Chuan LU Hong HAO Sheng-ju YAN You-sheng DANG Jie ZHENG Lei ZHANG Qing-hua HUO Zheng-hao*
    2016, (3):  409-414.  doi: 10.16098/j.issn.0529-1356.2016.03.021
    Abstract ( )  

    Objective The ratio of the length of the second finger to the fourth finger (2D∶4D) in humans is considered as a putative marker of prenatal exposure to androgen. Some studies have reported the relationship between CAG number and the ratio of 2D∶4D, but their results were variable. Here, we investigated the effect of CAG/GGN repeat polymorphisms in androgen receptor(AR)on 2D∶4D in Chinese people, and we wanted to investigate a suitable statistical method to find the real relationship between AR CAG/GGN repeat polymorphisms and the 2D∶4D ratio. Methods We examined 294 males and 391 females. The CAG and GGN repeats of the AR gene were analyzed on ABI 3730 DNA analyzer, using Peak Scanner software V1.0 to ascertain the size of AR alleles. The 2D∶4D ratio was standardized by x ±2s. We used the quartile method to choose the lower (Q1) and higher (Q3) 2D∶4D ratio. Pearson’s product moment correlation coefficients were used to find the relationship between AR CAG/GGN repeat polymorphisms and the 2D∶4D ratio. Results Females showed significantly higher 2D∶4D ratio than males in the right, and mean 2D∶4D ratio of both hards but not in the DR-L 2D∶4D ratio. There was no relationship between AR CAG/GGN alleles and right、left 2D∶4D of males (P>0.05),but CAG alleles were positively related to Q1 DR-L, Q3 Mean 2D∶4D ratio of males (r=0.280,P<0.05,r=0.274,P<0.05). The shorter CAG alleles were positively related to Q3 Mean 2D∶4D ratio of females (r=0.337,P<0.05),the longer CAG alleles were positively related to Q3 right hand and DR-L 2D∶4D ratio of females (r=0.238,P<0.05;r=0.175, P<0.05),the Mean CAG alleles were positively related to Q3 Mean 2D∶4D ratio of females (r=0.236, P P<0.05), the longer GGN alleles were positively related to Q1 right hand 2D∶4D ratio of females (r=0.204, P<0.05),the Mean GGN alleles were positively related to Q3 Mean 2D∶4D ratio of females (r=0.225, P<0.05). The longer CAG alleles were positively related to right hand 2D∶4D ratio of females. The longer GGN alleles were positively related to right hand 2D∶4D ratio of females. Conclusion 2D∶4D ratio may be used as a simple proxy for AR variation. Combine using the quartile method and Pearson’s product moment correlation coefficients may be a better way to find the relationship between AR variation and 2D∶4D.

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    A method of studying the related gene function of spinal cord neural stem cells based onin ovo electroporation for chicken embryos
    YU Ya-nan LIU Yan-li YANG Ci-qing ZHANG Bi-chao XU Zhen-ping LIN Jun-tang*
    2016, (3):  415-420.  doi: 10.16098/j.issn.0529-1356.2016.03.022
    Abstract ( )  

    Objective To develop a method of studying the related gene function of the chick spinal cord neural stem cells (NSCs) based onin ovo electroporation for chicken embryos. Methods The surface markers of the spinal cord NSCs were tested by RT-PCR at the chick different developmental stages. Subsequently, the pCAGGS-GFP plasmid was transfected into the chick spinal cord using in ovoelectroporation when the fertilized eggs were cultured for E2.5-E3; GFP-positive chick embryos were collected at E6, five samples in each group at least. The specimens were either transversely sectioned or undergone “open-book” processing. The commissural axons were observed under a fluorescent microscope. Finally, the spinal cords were isolated under the stereomicroscope, and the NSCs were cultured in 37℃ and 5%CO2 condition after trypsin digest. Results RT-PCR results showed that the NSCs surface markers were expressed in chick spinal cords; the subsequent results of transverse sections and open-book processing collectively showed that GFP labeling commissural axons crossed the midline region through the floor plate to the contralateral side of the spinal cord. The GFP labeling cells isolated from spinal cords exhibited typical morphological characteristics of NSCs, and generated neuritis when cultured in vitro. Conclusion A new method is successfully established, which could be used to study the gene function of the chick spinal cord NSCs based onin ovo electroporation.

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    Architecture of the lymphatic vessel revealed with the whole-mount immunostaining
    WANG Hai-jie * TAN Yu-zhen Pober Jordan S
    2016, (3):  421-424.  doi: 10.16098/j.issn.0529-1356.2016.03.023
    Abstract ( )  

    Objective To reveal lymphatic vessels in the tissue with the whole-mount immunostaining and compare this method with indirect injection and section immunostaining methods. Methods The lymphatic vessels in the capsule of the newborn liver were indirectly injected with Prussian blue; the architecture of the superficial hepatic lymphatic vessels was viewed. Drainage of the vessels was analyzed with collodion sections. Human skin of the femoral region was removed, and the distribution of the lymphatic vessels in paraffin and frozen sections was determined. After the skin of the ear and back, diaphragm and small intestine were removed from rats, the tissue pieces were stained with immunofluorescence, and the architecture of the lymphatic vessels in the tissue was examined.
    Results Indirect injection of Prussian blue revealed the architectures and drainage of the lymphatic vessels, while staining of the lymphatic vessels was short of specificity. With immunostaining, the tissue sections showed the distribution of the lymphatic vessels. However, the density of the lymphatic vessels was not analyzed accurately in this method. Whole-mount immunostaining of the tissues was desirable. The blind ends of the lymphatic capillaries, lymphatic capillary network and lymphatic plexus were clear. Conclusion The architecture of the intact lymphatic vessels in the tissue may be demonstrated well with the whole-mount immunostaining.

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    The regulatory effect of microRNAs on peripheral nerve regeneration
    2016, (3):  425-428. 
    Abstract ( )  

    microRNAs (miRNAs) are a class of highly conserved endogenous non-coding RNAs. Emerging studies demonstrate that miRNAs play important roles in many physiological and pathological processes. Following peripheral nerve injury, the expression levels of numerous miRNAs change significantly. Differentially expressed miRNAs negatively regulate their target genes, and thus affect the regeneration and reconstruction of injured peripheral nerve. In the current review, the authors discuss the regulatory effect of miRNAs on neuron, Schwann cells, and denervated muscles. A full cognition of these differentially expressed miRNAs benefits the understanding of the intrinsic molecular mechanisms underlying peripheral nerve regeneration and may provide a new strategy for the clinical application of miRNAs.

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    Review on optineurin mutation in neurodegeneration diseases
    LIU Hui SHEN Jing-ling*
    2016, (3):  429-432.  doi: 10.16098/j.issn.0529-1356.2016.03.025
    Abstract ( )  

    Optineurin (OPTN) is often considered as a pathological marker protein of Primary Open Angle Glaucoma (POAG) and Amyotrophic Lateral Sclerosis (ALS).OPTN participates in various cellular functions, such as regulation of post-Golgi membrane trafficking, secretion, autophagy of intracellular pathogen, antiviral innate immune response, regulation of mitosis,gene expression and NF-κB pathway. In the OPTN mutant ALS patient, the damaged motor neurons show cytoplasm OPTN ubiquitination aggregation, protein inclusions formation and co-locolized with ALS-interacting SOD1、TDP43、FUS proteins. In this review, we focus on recent studies on OPTN mutation in amyotrophic lateral sclerosis and its pathogenesis, pathology and treatment.

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