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    2014, Volume 45 Issue 6
    06 December 2012
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    Decrease of the glial and fibrotic scars tissue formation by degrading fibrinogen after traumatic brain injury
    BAI Dan LI Dan LIU Nan PEI Dan LI Hong-peng*
    2014, 45 (6):  729-734.  doi: 10.3969/j.issn.0529-1356.2014.06.001
    Abstract ( )  

    Objective To investigate probability of reducing formation of glial and fibrotic scar by degrading fibrinogen after traumatic brain injury. Methods The nigrostiatal dopaminergic pathway was unilaterally transected in 8-weeks-old Kunming mouse according to the method of Kawano et al. Adult male mice were anesthetized and transferred to a stereotaxic frame. A small oblong hole at the right rear of the bregma at 1.5mm and at a depth of 6.0 mm from the surface of the brain was made with a dental drill.. The blade was slowly pulled out, bleeding was stopped and the incision was sutured. Twenty four Kunming mice were randomly divided into control group and experimental group. For three consecutive days, experimental group mice were injected with batroxobin at 1 hour after operation. The mice brains were obtained at 4days, 7days and 14days after brain injury. Immunohistochemical localization of fibrotic scar and glial scar were examined by using the antibodies collagenⅣ(ColⅣ) and GFAP. The formation of scar tissue was observed by double immunofluorescent staining. Results After 4 days injury, the lesion site of the control group appeared ColⅣ deposition,which was surrounded by glial limitans formed by reactive astrocytes. Compared with the fibrotic scar of the lesion center 7days after injury, more obvious ColⅣ deposition was found at 14days than 7days after injury, and the lesion center was surrounded by the reactive astrocytes. However, the glial limitans were not found around the lesion center in the experimental group and fibrotic scar was almost disappeared, but the reactive astrocytes surrounded the lesion center. Double immunofluorescent staining demonstrated that there was FN immunoreactivity deposition in the lesion site of control groups forming fibrotic scar, while the experimental group reduced the deposition of FN significantly 7days after injury, and almost eliminated it at 14days after injury. Both groups had the GFAP positive cells surrounded the lesion site. Conclusion After traumatic brain injury, injection of batroxobin could decrease the formations of the glial scar and fibrotic scar by degrading fibrinogen.

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    Alteration of microglia and vasculature in the developing Tg2576 transgenic mouse
    LIU Kai NIU Yan-li LI Jin-ju WU Ping* DENG Jin-bo*
    2014, 45 (6):  735-741.  doi: 10.3969/j.issn.0529-1356.2014.06.002
    Abstract ( )  

    Objective Our aim is to study the alteration of microglia and vasculature in the developing hippocampus of Tg2576 transgenic mice. Methods Tg2576 transgenic mice from postnatal day 0 (P0) to P360 were used for Iba1 and NeuN immunohistochemistry, TdT-mediated dUTP nick-end labeling (TUNEL), ink perfusion and RT-PCR analysis. Results From P180, the density of Iba1-positive cells in CA1 area of the Tg2576 transgenic mouse was significantly higher than that of the wild type mouse. The vasculature (volumetric density) of the transgenic mouse at P360 was significantly less than in the wild-type mouse (P<0.01). RT-PCR results also showed that the activity of microglia was enhanced in the AD model. Conclusion The onset and development of Alzheimer disease is correlated to the alteration of microglia and vasculature in hippocampus.

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    Tissue affinity of bone marrow mesenchymal stem cell and the therapeutic transplantation for neurodegenerative diseases
    WEN Ya-nan NIU Yan-li HU Lei FU Xing LI Jin-ju WU Ping* DENG Jin-bo*
    2014, 45 (6):  742-748.  doi: 10.3969/j.issn.0529-1356.2014.06.003
    Abstract ( )  

    Objective To study the affinity of bone marreow mesenchymal stem cells (BMSCs) to various tissues and the possibility for therapeutic transplantation to cure neurodegenerative diseases with BMSCs. Methods The hippocampal slices, liver slices and lung slices were co-cultured with BMSCs to investigate BMSCs affinity to the tissues above, and immunohistochemistry (NeuN and GFAP) was carried out to understand the differentiation of BMSCs into neurons and glial cells under microenvironment of hippocampal slices, liver slices and lung slices. Results BMSCs had a highest affinity with hippocampal slices in contrast with the slices of liver and lung. The BMSCs’ temporal affinity on hippocampal slice was observed as well. At P7, BMSCs had the highest affinity with hippocampus, but there was a time-related decrease afterward. Differentiation of BMSCs occurred on hippocampal slices of both WT and Tg mice. Under the induction of hippocampal microenvironment, BMSCs differentiated into nervous cells and glial cells, but there was no significance between WT and Tg mouse. Conclusion BMSCs may provide an opportunity for the therapeutic transplantation to cure the neurodegenerative diseases and brain injuries. The time-related decrease of cell adhesion to hippocampus shows that the early stem cell transplantation is necessary.

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    Rhynchophylline attenuates cerebral vasospasm after subarachnoid hemorrhage in rats
    SUN Juan ZHANG Yan XU Ting YANG Lei ZHOU Chang-man LI Jian-hua* YAN Jun-hao*
    2014, 45 (6):  749-754.  doi: 10.3969/j.issn.0529-1356.2014.06.004
    Abstract ( )  

    Objective To investigate the effects of rhynchophylline (Rhy) on alleviating cerebral vasospasm (CVS) after subarachnoid hemorrhage (SAH). Methods SAH model was established in adult male SD rats by using endovascular perforation method. The Rhy was immediately applied by intraperitoneal injection after SAH. At 24hours after SAH, the diameter and vessel wall thickness of basilar artery (BA) were measured by magnetic resonance imaging (MRI) and/or HE staining. The levels of phosphorylated p38 mitogen-activated protein kinase(p-p38MAPK), p-p53 and cleaved-Caspase-3 in BA were determined by Western blotting and immunofluorescence. Results After Rhy treatment, the diameter of BA was significantly increased and the thickness of vessel wall was markedly decreased (P<0.05),Additionally, the levels of p-p38MAPK, p-p53 and cleaved-Caspase-3 in BA were significantly reduced by Rhy treatment after SAH(P<0.05). Rhy can reduce the expression of BA p-p38 MAPK, p-p53, cleaved-Caspase-3 apoptosis protein in endothelial cell by immunofluorescence compared with SAH+NS group. Conclusion Rhy can attenuate CVS severty after SAH partly through eudothelial cell appotosis via p38MAPK-p53-Caspase-3 pathway.

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    Role and mechanism of extracellular signal-regulated kinase/miR-133b pathway in methamphetamine-induced neurotoxicity in PC12 cells
    LIU Hai-li ZHU De-xiao WU Jin-tao YUE Qing-wei WANG Hui LI Ze-yan LI Gui-bao LIU Zeng-xun ZHANG Jing SUN Jin-hao*
    2014, 45 (6):  755-760.  doi: 10.3969/j.issn.0529-1356.2014.06.005
    Abstract ( )  

    Objective To investigate the expression of phosphorylated extracellular signal-regulated kinase (p-ERK) and miR-133b and their regulation mechanism in methamphetamine (MA)-induced neurotoxicity. Methods PC12 cells, used as models for neuronal cellsin vitro, were treated with MA with different concentrations, and the morphological changes were observed using an inverted microscope. MTT assay was used to observe the cell viability and determine the optimal MA concentration for cellular damage. Reactive oxygen species (ROS) level was measured with flow cytometry (FCM) by staining hepatocytes with DCFH-DA. Western blotting was used to detect the changes in the expressions of ERK1/2 and activated p-ERK1/2, and realtime quantitative PCR (Real-time PCR) was performed for miR-133b. To further analyze the ERK/miR-133b molecular pathway, U0126 was added to inhibit ERK phosphorylation to detect the changes of miR-133b. Results PC12 cells were damaged under the MA treatment of all concentrations. However, when exposed to 800μmol/L MA, the morphological changes of the cells were most significant, resulting in rounded cell bodies with dendrite disruptions and disappearance of cell reticular formations. Also, cell viability decreased significantly as shown by MTT, with increased levels of ROS and p-ERK and decreased expression of miR-133b. Applying U0126(10μmol/L), which inhibits ERK phosphorylation, before inducing cell damage with MA resulted in cells with increased expression of miR-133b, decreased level of ROS, increased cell viability, and thus weaker damage compared to the MA-induce-only group.
    Conclusion MA can induce cytotoxicity in the PC12 cells by up-regulating ERK phosphorylation and down-regulating miR-133b expression. U0126 can regulate the down-regulation of miR-133b by inhibiting ERK phosphorylation. Therefore, it is possible that the down-regulation of miR-133b in MA-induced neural toxicity is achieved by ERK phosphorylation.

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    Changes of tumor necrosis factor receptor-associated factor 6 expression in denervated tibialis anterior muscle and soleus muscle of rats
    SUN Hua-lin CHEN Yan-fei QIU Jia-ying WANG Chao GU Xiao-song DING Fei*
    2014, 45 (6):  761-767.  doi: 10.3969/j.issn.0529-1356.2014.06.006
    Abstract ( )  

    Objective To explore differentially expressed proteins between tibialis anterior muscle and soleus muscle after denervation by proteomics, and to analyze the changes of tumor necrosis factor receptor-associated factor 6(TRAF6)expression and its significance between tibialis anterior muscle and soleus muscle after denervation. Methods Rat models were prepared by cutting sciatic nerve. The protein expression changes between tibialis anterior muscle and soleus muscle after denervation were analyzed by isobaric tags for relative and absolute quantitation (iTRAQ) quantitative proteomics methods. The differentially expressed proteins were compared between tibialis anterior muscle and soleus muscle to screen key proteins regulating muscle atrophy. The expression was verified by Western blotting, and the biological role of key proteins was further validated in vitro. Results The atrophy of slow-twitch soleus muscle was faster than fast-twitch tibialis anterior muscle after denervation(P<0.05,n=20). There were 30 proteins which displayed different expression between tibialis anterior muscle and soleus muscle. These differentially expressed proteins included metabolic-related proteins, chaperones, contractile proteins and signaling molecules. Among the including signaling molecules, TRAF6 displayed a significant increase in slow-twitch soleus muscle compared to fast-twitch tibialis anterior muscle after denervation, which was confirmed by Western blotting(P<0.05,n=6). Muscle ring-finger protein-1(MuRF1)and muscle atrophy F-box(MAFbx), target genes of TRAF6, also showed a markly increase in slow-twitch soleus muscle compared to fast-twitch tibialis anterior muscle after denervation(P<0.05,n=6). In order to further investigate the impact of TRAF6 in muscle atrophy, TRAF6 siRNA and control siRNA were tranfected into myotubes, which was then treated by dexamethasone. The results showed that the diameter of myotubes transfected with TRAF6 siRNA was significantly larger than that in the myotubes transfected with control siRNA(P<0.05,n=6). The expression of TRAF6, MuRF1 and MAFBx in TRAF6 siRNA transfection group were significantly lower than that in the control siRNA transfected group(P<0.05,n=6), suggesting that TRAF6 siRNA effectively inhibited the expression of the target gene TRAF6, also inhibited the expression of downstream its target genes of MuRF1 and MAFBx. Conclusion TRAF6 displays a significant increase in slow-twitch soleus muscle compared to fast-twitch tibialis anterior muscle after denervation. Inhibition of TRAF6 alleviates myotube atrophy induced by dexamethasone. Therefore, we speculate that the atrophy of slow-twitch muscle is faster than fast-twitch muscle after denervation, which is possible caused by the higher expression of TRAF6 in slow-twitch muscle after denervation.

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    Expression of the autophagy-related protein in hippocampus in a rat model of autism
    CHEN Yun-hua LI Hong-wei TONG Xue-tao WEN Min*
    2014, 45 (6):  768-772.  doi: 10.3969/j.issn.0529-1356.2014.06.007
    Abstract ( )  

    Objective To investigate the effect of autophagy in the rat model of autism. Methods Twenty female Wistar rats (weight 250-260g) were mated with 10 male (weight 280-290g) overnight, and in the morning when spermatozoa were found it was designated as the first day of gestation. Females received a single intraperitoneal injection of sodium valproate on the 12.5 day after conception, and control females were injected with physiological saline at the same time. The offspring of valproate-treated females were model group, and the offspring of physiological saline -treated females were control group. The self-grooming test and three-chambered social test were used to confirm whether the autism model were successful. Western blotting was used to detect the expressed level of LC3-Ⅱ, Beclin 1 and P62 in hippocampal of eatch group. Results 1.The animal model of autism was successfully established. Compared with the control group, the cumulative self-grooming time was significantly increased(P<0.05), the sociability and preference for social novelty were significantly decreased(P<0.05). 2. Compared with normal control group, the model group significantly decreased the expression level of LC3-Ⅱ(P<0.05)and Beclin 1(P<0.05)in hippocampal. Meanwhile the expression level of P62 was significantly increased(P<0.05). Conclusion The present findings confirmed that the autophagy is inhibitied in hippocampus of autism, which suggests modulation of autophagy might be a potential therapeutic strategy for autism.

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    Scutellaria baicalensis stem leaf flavonoids exerts protective effect on ischemia reperfusion injury of microvessel and brain blood barrier in cerebral hippocampus
    KONG Xiang-yu KONG Wei MIAO Guang-xin JI Hai-ru CHEN Meng ZHENG Xiao-ying ZHAO Shu-min*
    2014, 45 (6):  773-778.  doi: 10.3969/j.issn.0529-1356.2014.06.008
    Abstract ( )  

    Objective To explore the preventive protective effects on ischemia reperfusion injury of microvessels and blood brain barrier(BBB) in cerebral hippocampus with scutellaria baicalensis stem leaf total flavonoids (SSTF). Methods Ninety SD rats were assigned randomly into sham operation, ischemia reperfusion, and SSTF pretreated groups. The SSTF group rats were administrated with SSTF intragastrically one week before animal model-making. Low, medium and high dose groups were administrated with 50, 100, 200 mg/(kg ·d), respectively, for 7 days. The model group and SSTF group animals were remodelled into focal ischemia reperfusion models. Changes of neurological defects were evaluated. Postoperatively, water contents and microvascular permeability in brain tissue were detected with wet weight methods and Evans blue (EB) methods. The microvessel density (MVD) and microvessel area ratio(MVA) were observed with tannic acid-ferric chloride mordant dyeing methods (TA-Fe). The change of aquaporin 4(AQP4) mRNA expression was assessed by Real-Time PCR. The ultrastructural integrity of the blood brain barrier was observed under an electron microscope. Results The neurological function defect scale, water content in brain tissue and microvascular permeability in each SSTF group were significantly reduced compared with IR groups(P<0.01,P<0.05. The MVD and MVA were obviously increased (P<0.01), the expression of AQP4 was reduced significantly (P<0.01), the BBB injuries were alleviated in various degrees, the endothelial cells swelling gradually disappeared, the unloosed tight junction also gradually reclosed and recovered into a normal status, the basal membrane gradually returned into continuous and clear, and the gliacyte cytoplasm dissolving in pedal plate was alleviated and gradually turned close to normal status. While the above alleviate improvements in SSTF M and SSTF H groups were more conspicuous than that in SSTF L group (P<0.01), there was no significant difference between SSTF M and H groups (P>0.05). Conclusion Cerebral ischemia reperfusion can lead the hippocampus microvessel and blood brain barrier to be injured, and SSTF intervention can exert preventive protective effects on cerebral hippocampus tissue. The functional mechanism may carry out effectively by increasing the number of microvascular recanalization, by maintaining the microvessel and brain blood barrier to be in intact status and in normal function, as well as by relieving cerebral edema. It is proved that the optimal effective prevention dose of SSTF is 100mg/(kg·d).

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    Expression and significance of arrestin beta 1 in patients with glioma
    WEN Pu-shuai*GAO Jing LI Wen-hui XI Huan-jiu
    2014, 45 (6):  779-784.  doi: 10.3969/j.issn.0529-1356.2014.06.009
    Abstract ( )  

    Objective To detect the expression levels of arrestin beta 1 (ARRB1) in patients with glioma, and assess its clinical significance. Methods The expression levels of ARRB1 in glioma cell lines were detected by RT-PCR and Western blot; ARRB1 mRNA expression in glioma was analyzed from Oncomine database and Project Betastasis platform; the expression of ARRB1 in glioma and peritumoral brain tissue was detected by immunohistochemistry; the association of ARRB1 expression levels with the prognosis of patients was analyzed by Kaplan Meier method. Results Compared with normal brain tissue and cell line HEK293, the mRNA and protein levels of ARRB1 in glioma cell lines were reduced; Oncomine database showed ARRB1 mRNA levels in multiple glioma microarray were significantly lower than the normal control group (P<0.001), specially in highly malignant glioblastoma. Immunohistochemistry data showed that ARRB1 expression in glioma tissues was reduced compared with the peritumoral brain tissue, and the expression of ARRB1 in Ⅲ-Ⅳ stage of gliomas was reduced more significantly than that in stage Ⅰ-Ⅱ glioma. In addition, Kaplan-Meier survival curves showed that the expression level of ARRB1 in glioma was correlated with patients survival (P<0.05). Conclusion Reduced ARRB1 expression level in glioma could be used to evaluate the clinical and pathological features of glioma, and the ARRB1 can be used a biomarker to judge the prognosis of patients with glioma.

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    Effect of angelica polysaccharide on hematopiesis in aging rats and its biological mechanism
    ZHANG Yan-yan LI Jing JIA Dao-yong ZHANG Meng-si XU Chun-yan JING Peng-wei LI Cheng-peng WANG Lu WANG Shun-he WANG Ya-ping*
    2014, 45 (6):  785-792.  doi: 10.3969/j.issn.0529-1356.2014.06.010
    Abstract ( )  

    Objective To explore the effect and mechanism of angelica sinensis polysaccharide(ASP)on hematopiesis in aging rats and to provide theoretical and experimental evidences for effective natural medicine of antiaging or protecting hematopoietic function. Methods Male SD rats(n=40)aging from 6 to 8 weeks old were randomly divided into normal group(NG, n=10),ASP normal group(ANG, n=10), aging model group(AMG, n=10) and ASP aging model group(AAMG, n=10). After 2 days of the treatment, the eyeball blood was collected for the peripheral blood routine test; The femur was taken to count the number of each femur bone marrow mononuclear cells (BMNCs). The proliferation of BMNCs was detected by CCK-8. The distribution of cell cycles and reactive oxygen species (ROS) levels were analyzed by flow cytometry(FCM). The capability of colony formation was examined by CFU-Mix cultivation. The ratio of the SA-β-Gal staining positive BMNCs was counted. The total antioxidant capacity of the cell was detected by enzymatic assay. The aging related proteins of P53 and P21 were detected by Western blotting. Results Compared with the aging model group, in the ASP aging model group,the decrease in the amounts of the peripheral blood RBC, PLT and WBC was evidently inhibited; the proliferation of the BMNCs was enhanced;the colony formation of CFU-Mix was markedly increased; the percentage of SA-β-Gal, the ratio of G1 stages and the product of ROS positive cells were significantly reduced; the ratio of S stages was markedly promoted; the total antioxidant capacity of the cell was significantly promoted; the expression of the aging related protein of P53 and P21 was evidently down-regulated.
    Conclusion ASP can protect hematopoietic function from decline by antagonize D-galactose that induced oxidative stress, suggesting the mechanism of ASP protecting hematopiesis may regulate p53/p21 pathway.

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    Effects of curcumin on proliferation and adipogenic differentiation of porcine bone marrow mesenchymal stem cells
    ZHANG Qing-mei LI Fang-zheng JIANG Zhong-ling SUN Dong-xing LI Dong-jian SONG Xue-xiong*
    2014, 45 (6):  793-799.  doi: 10.3969/j.issn.0529-1356.2014.06.011
    Abstract ( )  

    Objective To investigate the effects and mechanisms of curcumin on proliferation and adipogenic differentiation of porcine bone marrow mesenchymal stem cells (BMSCs). Methods The porcine BMSCs were cultured and differentiated with cell culture and adipegenic differentiation medium containing different concentrations of curcumin, the effects of curcumin on proliferation of porcine BMSCs were detected by MTT, the effects of curcumin on adipogenic differentiation were analyzed by morphology and oil red O, and the expressions of peroxisome proliferator-activated receptor γ(PPARγ2) mRNA and lipoprotein lipase(LPL) mRNA were detected by Real-time PCR. Results When the porcine BMSCs were cultured with curcumin, 1μmol/L curcumin at the third days of culture and 5, 10μmol/L curcumin at the fifth days of culture significantly promoted cell proliferation (P<0.05), but curcumin concentration higher than 15μmol/L inhibited cell proliferation (P<0.05). When the porcine BMSCs were differentiated with curcumin, the different concentrations of curcumin significantly inhibited adipogenic differentiation (P<0.05). The inhibition rate of 0.5μmol/L curcumin was 28.3% and 20μmol/L curcumin was 71.24%. When the cells were treated with 20μmol/L curcumin and differentiated in 5, 10 and 15 days, the inhibition rates of expressions of PPARγ2 and LPL mRNA were 19.11%, 49.52%, 76.76% and 37.58%, 49.32%, 74.70% respectively.
    Conclusion The proper concentrations of curcumin were able to promote the proliferation and inhibit adipogenic differentiation of porcine BMSCs, suggesting that curcumin may play an important role in the regulation of adipocyte differentiation.

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    Molecular mechanism on nonalcoholic fatty liver disease by liraglutide regulation
    NIU Shi-wei WU Jun-zi LI Xiao-bo WANG Wen-lin LI Yan1* LI Shu-de*
    2014, 45 (6):  800-808.  doi: 10.3969/j.issn.0529-1356.2014.06.012
    Abstract ( )  

    Objective To investigate the effect of liraglutide on the reaction of inflammation and antioxidant capacity on the model of nonalcoholic fatty liver disease (NAFLD) rats and study the regulation of liraglutide to the expression of lipid metabolism enzyme. Methods Seventy SD rats, 25 SD rat were given normal cliet, the others were induced to produc NAFLD by high glucose and high fat diet by gavage for 12 weeks, five rats were killed randomly from control group and model group respectly. After the successful establishment of NAFLD model,the rats were randomly divided into the control group (n=20), the model group (n=20) and the liraglutide treatment group (n=20).The liraglutide treatment group was given liraglutide 60μg/(kg·d) by subcutaneous injection, while the model group received normal saline 1ml/(kg·d). The rats were killed 4 week or 8 weeks after the liraglutide injection, 50% from each group at each time point, so as to observe the changes of inflammatory factors, oxidant factors, antioxidant factors, liver function and lipids in the plasma, and to detect the liver morphological changes and the mRNA expressions of sterol regulatory element binding transcription factor -1c (SREBF-1c) and acetyl coenzyme A carboxylase alpha (ACCα) by the reverse transcription-PCR(RT-PCR). The protein expressions of SREBF-1c and ACCα were detected by the immunohistochemistry and Western blotting. Results Compared with the model group, C-reactive protein (CRP), interleukin(IL)-1α, IL-1β, IL-6, gumor necrosis facto-α(TNF-α), malondialdehyde(MDA), aspartateamino trans ferase(AST), alanineamino transferase(ALT), triglyceride(TG), total cholesterol(TC) and low-density lipoprotein(LDL-C) contents in the plasma of the rats with the liraglutide treatment were reduced after 4 weeks(P<0.05), with superoxide dismutase(SOD), total antioxidant capacity(T-AOC) and glutathione (GSH) being increased (P<0.05), liver fat particle deposition reduced and the expressions of mRNA and protein of SREBF-1c and ACC α in the liver tissue regulated downward(P<0.05). After the liraglutide treatment for 8 weeks, the above indicators were improved were more obviously. Conclusion Liraglutide can reduce the inflammatory reaction in NAFLD and improve the antioxidant capacity as well as the liver function. It can also reduce the generation of lipid by down-regulating the expressions of SREBF-1c and ACCα in liver tissue possibly, and alleviate the liver fatty degeneration. With the increase of treatment time, the effect of liraglutide on the prevention and treatment of NAFLD becomes more obvious.

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    Influence of annexin A7 knockdown on expression of Bcl-2 and Bax of HepG2 cells
    WANG Xiao-jie LIANG Xiu-jun LI Xin*
    2014, 45 (6):  809-813.  doi: 10.3969/j.issn.0529-1356.2014.06.013
    Abstract ( )  

    Objective To investigate the influence of annexin A7 knockdown on apoptosis and expression of Bcl-2 and Bax of HepG2 cells. Methods HepG2 cells cultured in six-well plates were divided into 3 groups, siRNA interference group, negative control group and blank control group. The siRNA interference group was transfected with the siRNA which target annexin A7, the negative control group was transfected with negative siRNA, in the blank control group only transfection reagent was added. Western blotting was used to identify the siRNA targeted to annexin A7 can highly surpress annexin A7 expression in 48h after transfection. Flow cytometry was used to detect the apoptotic rate. Immunohistochemistry and Western blotting were applied to observe the expression of Bcl-2 and Bax protein; RT-PCR was performed to detect the expression of Bcl-2 and Bax mRNA. Results The apoptotic rate increased and the expression of Bcl-2 decreased significantly compared with the negative control group and the blank control group(P<0.05), while the expression of Bax protein and mRNA had no significant difference when compared with the negative control group and the blank control group(P>0.05). Conclusion Annexin A7 knockdown can promote apoptosis of HepG2 cells, reduce the ratio of Bcl-2 and Bax.

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    Applied anatomy of intrusive operations of sacral cyst
    YAN Jia FAN Yan LIU Cheng-xing BAI Yong ZHANG Xiang LIU Zong-liang BA Ying-chun*
    2014, 45 (6):  814-818.  doi: 10.3969/j.issn.0529-1356.2014.06.014
    Abstract ( )  

    Objective To identify sacral hiatus through the sacral bony landmarks, and to clarify its positional relationship with the sacral ganglion, and to provide anatomical guidance of the angle and depth of the needle for interventional operation through sacral hiatus. Methods Fifteen adult cadavers were included in this study. The distance and angle between the sacral hiatus and the sacral ganglion and the triangle which was formed by left and right posterior superior iliac spine and the midpoint of the sacral cornu were measured. Various anatomical measurements on the sagittal section were conducted. The angle and depth of the inserted puncture needle were recorded. Results Average distance from the midpoint of the sacral cornu to the end of the coccyx was (5.73 ± 0.81) cm. Both sides of the posterior superior iliac spine and the midpoint of sacral cornu constituted an isosceles triangle. The average depression angle of needle from the sacral hiatus was 60.4 degrees. Distance from the midpoint of both sides of the sacral hiatus to lower bound of the dural was greater than to the third sacral ganglion, and shorter than to the first and second midpoint of sacral ganglia. Conclusion This study summed up surface location of the sacral hiatus, clarified its body surface, depth, positional relationship with the sacral ganglion and the best angle for needle piercing. These findings can help clinicians perform safer diagnosis and treatment on the of the sides sacral cyst, and also provide a useful reference for caudal anesthesia, RF rhizotomy, and ultrasound examination.

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    Anatomy and modification of the middle window in the subilioinguinal approach
    XU Shun-li ZHAO Xue-song Lü Xiang WANG Yi-sheng*
    2014, 45 (6):  819-823.  doi: 10.3969/j.issn.0529-1356.2014.06.015
    Abstract ( )  

    Objective To provide the anatomical evidence to prevent vascular damage and lyphatical leakage during the subilioinguinal approach for acetabulum fracture. Methods Twenty human adult cadavers were dissected in the Anatomy Department of Xinxiang Medical University.The structures associated with the middle window of the subilioinguinal approach were observed. Results Inferior epigastric artery was derived from external iliac artery (57.5%) or femoral artery(17.5%), was (41.35±2.33)mm from pubic symphysis,and had a (40.23±5.23)° angle with external iliac artery.There was no difference between left and right sides,and no correlation with the width and length of pelvis. There was no fascia-like structure around the iliac blood vesses. The fascia-like structure was observed around the femoral blood vessels but it was easy to be separated from the surrounding structures. The angle between the origin of iliopectineal arch and the medial end of the inguinal ligament was sharp and at (15.20±6.79)°and filled by a thin membrane-like structure. The medial and inferior part of the iliac fascia was thick and had no correlation with the width and length of pelvis. Conclusion Blood vessel and lymphatic vessel are prone to injury in the subinguinal and inguinal approaches. Inferior part of iliac fascia and iliopectineal arch are compact, and thus the lateral window may pass through medially under the fascia iliaca.

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    Gastrodin inhibits lipopolysaccharide-induced autophagy in human umbilical vein endothelial cells
    XIE Ya-nan YANG Ping WEN Fei SUN Lin CHEN Yuan-li* LU Di*
    2014, 45 (6):  824-829.  doi: 10.3969/j.issn.0529-1356.2014.06.016
    Abstract ( )  

    Objective To investigate the regulation mechanism of gastrodin (Gas) on atutophagy of human umbilical vein endothelial cells(HUVECs)stimulated with lipopolysaccharide (LPS). Methods Gas at 5, 10, 20 and 50μmol/L was added 1h before 1mg/L LPS stimulation, after 12h incubation. The protein and mRNA expression of Beclin-1, ATG5 and LC3 were detected by Western blottingting and RT-PCR respectively. The autophagosome was observed under fluorescent microscope by monodansylcadaverine (MDC) staining. Cell viability rate was measured by MTT analysis. Results Autophagy was induced in HUVECs by LPS-stimulation. The protein and mRNA expression of Beclin-1, ATG5 and LC3 were significantly increased,which was in a time-dependent manner. Compared with LPS-stimulated group, pertreatment with Gas inhibited the expression of autophagy in dose-dependent manner and the cell viability was effectively increased. Conclusion Autophagy was induced in HUVECs by LPS-stimulation. Gas may increase the viability rate of HUVECs through inhibiting the expression of autophagy.

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    Expressions of the RECK, matrix metalloproteinases-9 and vascular endothelial/growth factor in placentas of preeclamptic patients
    HUA Mao-fang * LIU Xiao-yun LI SHan-feng
    2014, 45 (6):  830-834.  doi: 10.3969/j.issn.0529-1356.2014.06.017
    Abstract ( )  

    Objective To investigate the expression of reversion-inducing cysteine-rich protein with reversion-inducing-cysteine-rich protein with kazal motifs (RECK), matrix metalloproteinase-9 (MMP-9) as well as vascular endothelial growth factor (VEGF) genes in placenta of preeclamptic patients, and discuss the regulation of these genes in invasion of placenta trophoblasts. Methods Reverse transcription polymerase chain reaction (RT-PCR), Western blotting and immunohistochemistry were employed in the detection of expression of RECK gene, MMP-9 and VEGF in placenta tissues from 120 gravidae with full-term pregnancy undergoing cesarean section, i.e., normal pregnancy group (n=30), mild preeclampsia group (n=30), moderate preeclampsia group (n=30) and severe preeclampsia group (n=30). Results The mRNA expression level of MMP-9 and VEGF in three preeclampsia groups were all significantly lower (P<0.05) than those in normal pregnancy group. The expression level of RECK mRNA in the severe preeclampsia group was significantly higher (P<0.05) than that in normal pregnancy group. The expression of MMP-9 and VEGF proteins in three preeclampsia groups were lower than those in normal pregnancy group with statistically significant differences (P<0.05),and the expression of RECK protein in moderate and severe preeclampsia groups were both significantly higher (P<0.05) than that in normal pregnancy group. Conclusion The expression level of RECK is negatively correlated with those of MMP-2 and MMP-9 in preeclamptic patients, which indicates that they are involved in regulating the process of shallow invasion of trophoblasts in placenta.

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    Relationship between cardiac function and angiogenesis after rat myocardial infarction
    ZUO Hong-bo LI Ci-xia GUO Zhi-kun*
    2014, 45 (6):  835-840.  doi: 10.3969/j.issn.0529-1356.2014.06.018
    Abstract ( )  

    Objective To explore the relationship between changes of heart function and angiogenesis after myocardial infarction, and provide morphological reference for clinical treatment of myocardial infarction. Methods Acute myocardiac infarction (AMI) model was established with Sprague-Dawley rats by ligation of the left anterior descending coronary artery.Cardiac function at different weeks after AMI was evaluated by echocardiography. The change of Ⅷ factor and vascular endothelial growth factor(VEGF)expression at the infarction edge in rats hearts at each week after AMI was detected by immunohistochemistry and Western blotting. Results The cardiac function of 1W after myocardial infarction was worst, cardiac function was improved during 2 weeks and 3weeks, but reduced at the 4th week; Ⅷ factor and VEGF in post-myocardial infarction 1 week, 2 weeks were highly expressed. Expression of both factors began to decrease at the 3rd week. At the 4th week, expression levels were close to normal. Conclusion The change of cardiac function after myocardial infarction is inconsistent with that of angiogenesis, early increased vascular density can not obviously improve cardiac function.

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    Differentiation of cardiomyocyte-like cells from human umbilical cord-derived mesenchymal stem cells induced by valsartan
    HE Wan-li LI Yong-hai YANG Jun-zheng LIN Jun-tang GUO Zhi-kun XU Zhen-ping*
    2014, 45 (6):  841-846.  doi: 10.3969/j.issn.0529-1356.2014.06.019
    Abstract ( )  

    Objective To explore the feasibility of differentiating the human umbilical cord-derived msenchymal stem cells (hUCMSCs) to myocardiocyte-like cells after induction with valsartan. Methods The hUCMSCs were induced by valsartan and cultured for 1 week, 2 weeks and 3 weeks respectively. The antibodies against cardiac troponin T (cTnT)and recombinant GATA binding protein 4 (GATA4)were used to identify the cells which were induced to differentiate by immunocytochemistry and immunofluorescence. The mRNA expressions of cardiac troponin I(cTnI), myocardial-related transcription factor and myocardial development-related genes were detected by RT-PCR to identify the feasibility of differentiating hUCMSCs and inducing capacity of valsartan. Results The morphology of normal hUCMSCs showed uniform fibrous and whirlpool-like shape. They became significantly smaller and short spindle shape after valsartan induction. Immunochemical and immunofluorescence results showed that the expression of cTnT and GATA4 protein in the induced group was positive and seldom in the control group. RT-PCR results showed that the mRNA expression of cardiac-related transcription factor Tbx5, GATA4 and Nkx2.5 was spontaneously and continuously in the hUCMSCs. They increased after being induced by valsartan 1 week and then declined. The mRNA expression of cTnI was detected at 3 weeks, but the atrial natriuretic polypeptide(ANP)and β-myosin heavy chain (β-MHC) were not detected. Conclusion The hUCMSCs have capacity to be induced to differentiate into cardiomyocyte-like cells by valsartan.

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    Variation in chin form of Chinese population
    DAI Cheng-ping LI Hai-jun*
    2014, 45 (6):  847-849.  doi: 10.3969/j.issn.0529-1356.2014.05.020
    Abstract ( )  

    Objective To understand the variation in the chin form of Chinese population. Methods This study was conducted on 364 human mandibles from China. Chin form is divided into 3 types: straight, round and protruding, and comparisons were made among different time periods. Results The “round” and “straight” were the two main chin forms in Neolithic age, while most were “Round” in Bronze-Iron age and present. The frequency of “straight” was the highest in Neolithic age, and the lowest in Bronze-Iron age. Conclusion The chin form varies in different time periods, which may be caused partly by the size variation of mandibles.

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    Variation of some mandibular traits related to masticatory muscles in Chinese population
    LI Hai-jun* DAI Cheng-ping
    2014, 45 (6):  850-854.  doi: 10.3969/j.issn.0529-1356.2014.06.021
    Abstract ( )  

    Objective To understand the variation of some Chinese non-metric mandibular traits related to masticatory muscle. Methods This study was conducted on 379 human mandibles from China. Mandibular traits was divided into several types, and then comparisons were made among different time periods. Results Tuberositas masseterica and tuberositas ptelygoidea were stronger in Neolithic Age than in the Bronze and Iron Age, and became weaker as human beings move into the modern era. Mental spine showed no big different among different time periods. Conclusion Masseter muscles and musculi pterygoideus internus may become weaker as time period goes by in Holocene, which may be caused partly by the decrease of masticatory force needed.

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    Analysis on regional differences of body surface area development of Chinese Han teenagers in rural areas
    GAO Guo-zhu* CHEN Li-wu
    2014, 45 (6):  855-860.  doi: 10.3969/j.issn.0529-1356.2014.06.022
    Abstract ( )  

    Objective To investigate the regional differences of the body surface area development of Chinese Han teenagers from areas. Methods This report was based on the data of height and weight of 107745(male 53857,female 53888)Chinese Han teenagers’ physique and health survey report in 2010 of 30 provinces and cities. It calculated the body surface came with Stevenson and DuBois’s formula. The 36 parameters of height, weight and body surface obtained from teenagers of 7-18 years old. It used varimax to factor analysis, average of the various parameters change to factor points and factors points to the samples Q cluster analysis (Ward’s method). Thirty samples of men and women were divided into several categories using the Cubic model fitting growth curve of the above body surface area. Results The three indicators(height, weight and body surface area) comprehensively summarized the differences of growth and development of different area groups. Both boys and girls were embraced into three categories. Boys were be divided into high and low development group. Girls were be divided into high, middle and low group. There were development differences among different groups. Conclusion There are both similarities and differences of body surface area development. As a whole the body surface area of Chinese Han teenagers in rural regions of China is lower from north to south and from east to west. The gap of the girl’s body surface area in China is reducing now, and the body surface of the middle-eastern and coastal provinces and cities has a tendency to go beyond the north’ body surface.

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    Glutamate release from isolated synaptoneurosomes from the mouse cortex
    JIN Guo-rong DING Cai-yun WANG Yi-min LIU Jian-rong WEI Hong-en*
    2014, 45 (6):  861-864.  doi: 10.3969/j.issn.0529-1356.2014.06.023
    Abstract ( )  

    Objective To detect the release of glutamate from the isolated synaptoneurosomes from the mouse cortex. Methods The mouse cortex was dissected, homogenized and filtered through three layers of 147 μm nylon meshes. The synaptoneurosomes were resuspended after centrifuged. An enzyme-linked fluorescence detection method was used to detect the glutamate release, which utilized glutamate dehydrogenase, and the reduction of nicotinamide adenine dinucleotide phosphate(NADP) to NADPH. The glutamate standard curve was set up and then the glutamate release from isolated synaptoneurosomes was tested and analyzed in three independent experiments. Results A good linear relationship was found between the glutamate concentrations and fluorescence values. The release of glutamate from synaptoneurosomes significantly increased after depolarization agent KCl added and gradually reached a plateau. Glutamate release was increased more when Triton X-100 was added. The inactive synaptoneurosomes after one freeze-thaw cycle had a different curve of glutamate release. Conclusion The release of excitatory neurotransmitter glutamate from synaptoneurosomes of the mouse cortex isolated using the improved method is well studied.

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    Research progress of biological function in integrin-interacting protein kindlin family
    YANG Mei CHEN Xi ZHANG Shui-wen ZHANG Hong-quan*
    2014, 45 (6):  865-869.  doi: 10.3969/j.issn.0529-1356.2014.05.024
    Abstract ( )  

    Integrin-interacting protein kindlin is a novel family of focal adhesion proteins consisting of three members, Kindlin-1, Kindlin-2 and Kindlin-3. Kindlins are highly conserved and belong to FERM-containing proteins. Present studies have showed there is a close relationship between Kindlin family and integrin activation and genetic diseases. In addition, Kindlin family is also a critical mediator for tumorigenesis. We review structural features, distribution and progress of biological functions of kindlin family in this paper.

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