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    2014, Volume 45 Issue 1
    06 February 2014
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    Aging related autofluorescence can be used as a marker of cell senescence
    ZHANG Jing YANG Lu-meng CHEN Xiao-chun*
    2014, 45 (1):  1-6.  doi: 10.3969/j.issn.0529-1356.2014.01.001
    Abstract ( )  

    Objective To describe the emission spectra of autofluorescence derived from lipofuscin in aged brain sections and to investigate how aging related autofluorescence is used reasonably and abolished properly. Methods Three months old and 24 months old C57BL/6J male and female mice were used in this study. Staining of senescenceassociated beta galatosidase and accumulation of lipofuscin were used to evaluate the degree of brain aging. Laser confocal microscopy was used to analyze the emission spectra of autofluorescence. Immunohistochemistry and immunofluorescence was used to identify astrocyte. Results Strong SA-β-Gal staining and autofluorescence and accumulation of lipofuscin appeared in the CA3 of hippocampus and the periventricular area of the third ventricle in 24 months old mice. The emission wavelength of aging related autofluorescence was between 500-565 nm. Astrocytes in periventricular area of the third ventricle were of not only strong SA-β-Gal staining, but also strong autofluorescence. Abolition of background autofluorescence from lipofuscin following immersion in autofluorescence eliminator reagent was observed in aged brain tissue. Conclusion Aging related autofluorescence can be used as a marker of cell senescence with a specific phenotype. Reduction of background autofluorescence from lipofuscin can improve the specificity of immunofluorescence detection in aged brain tissue samples.

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    Prenatal alcohol exposure inducing insulin resistance and stress injury in mouse hippocampi
    ZHOU Shu-fang GAO Lin HE Wei-ya Cui Zhan-jjun DENG Jin-bo*
    2014, 45 (1):  7-14.  doi: 10.3969/j.issn.0529-1356.2014.01.002
    Abstract ( )  

    Objective To investigate the relationship of prenatal alcohol exposure inducing insulin resistance and the stress injury in the mouse hippocampi.
    Methods C57BL/6J mice were used to establish the models of prenatal alcohol exposure with control group, moderate ethanol group and high ethanol group. The pups of different groups at postnatal day 7(P7), P14 and P30 were gathered for the measurements of fasting blood glucose and blood insulin. The insulin resistance index (HOM-IR) was calculated. The stress injury in hippocampal CA1 area was observed. For instance, the expression of stress injury proteins, c-Fos and NF-κB, of hippocampus at P7 and P14 was tested with immunofluorescent labeling and Western blotting to analysis. Results After prenatal alcohol exposure, fasting glucose and insulin resistance index increased with dose dependency and long term effect (P<0.05,n=90). After prenatal alcohol exposure, c-Fos and NF-κB positive cells increased in hippocampi with dose dependency and long term effect (P<0.05,n=90). The expression of c-Fos and NF-κB proteins with Western blotting in hippocampus was harmonized with the results of immunocytochemistry (P<0.05,n=30). Conclusion The prenatal alcohol exposure can induce pups insulin resistance, and the insulin resistance is probably caused by stress injury. In addition, the insulin resistance may be involved in the pathogenesis and brain injuries of fetal alcohol syndrome, through c-Fos and NF-κB pathway.

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    Abnormal expression of AMPA receptor in hippocampus of senescence accelerated mouse prone 8 correlated to synaptic plasticity
    FENG Min ZHANG Ying-jun LU Juan XIONG Yin-yi WU Qiao-feng YU Mei-ling LU Sheng-feng YU Shu-guang*
    2014, 45 (1):  15-19.  doi: 10.3969/j.issn.0529-1356.2014.01.003
    Abstract ( )  

    Objective To observe the expression differences of AMPA receptor in hippocampal neurons between SAMP8 and SAMR1 and to explore the pathogenesis of Alzheimer’S disease(AD). Methods Male SAMP8 mice of 10-month old were taken as animal model of AD and SAMR1 mice were taken as control group. Morris water maze(MWM), electron microscope and western blotting were used in this study. Results Comparing with the control group, the model group had significant learning impairment and memory retention deficits. The thickness of post synaptic density(PSD) and the curvature of the synaptic interface in the model group dicrease. The width of synaptic cleft in the model group increased. The expression of GluR2 in the model group dicreased. There were no significant differences in the expression of GluR1 between two groups. Conclusions Impaired synaptic plasticity mediated by pathological alteration of AMPA receptor in hippocampus mey be the pathogenesis underlying learning and memory disorders of AD.

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    Effect of epigallocatechin gallate on temporal and spatial profiles of neurons apoptosis and Caspase-3 protein expression after spinal cord injury in rats
    LIU Xin-ning DIAO Yao* LAN Zhen-he MA Jing GE Rui JIAO Ying ZHU Yue
    2014, 45 (1):  20-25.  doi: 10.3969/j.issn.0529-1356.2014.01.004
    Abstract ( )  

    Objective To evaluate the effect of epigallocatechin gallate(EGCG) on temporal and spatial profiles of neuron apoptosis and its associated Caaspase-3 protein expression after spinal cord injury in rats. Methods SD rats were randomly divided into three groups (25 rats each): sham-operated group, SCI group (normal saline solution intraperitoneal injection, immediately and 1 hour after SCI), and EGCG treatment groups (50mg/kg, intraperitoneal injection, immediately and 1 hour after SCI ). SCI model was made by clamps methods. The rats were sacrificed at 6, 12, 24 hours, 3 and 7days; 5 rats at each time point after injury. Spinal cord tissue in the injured area was harvested, sectioned 0-5.00 mm rostral from the epicenter post-injury and examined by cresyl violet (CV) staining and terminal-deoxynucleoitidyl transferase (TdT)-mediated deoxy uridine triphosphate-biotin nick end labeling (TUNEL) staining method and immunohistochemical method. Results At all the time points observed, CV-staining result showed that the neurons was not found in section 0-0.50 mm for SCI and EGCG group, the neurons was increased in EGCG group than in the SCI group in the sections 1.00-4.60 mm. The number of TUNEL positive neurons and Caspase-3 neurons were decreased in EGCG group than in SCI group in the section 1.05-3.05 mm and 1.10-3.10 mm rostral from the epicenter. Conclusion EGCG can decrease the expression of Caspase-3 and inhibit neuronal cell apoptosis in the rats after spinal cord injury on temporal and spatial profiles, which may play a significant neuroprotective role in the secondary SCI.

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    A semi-quantitative RT-PCR analysis for NeuroD mRNA expression in brain during rat development
    GUO Wei LIU Xiao-yan KE Li-ning LI Yu-yu LIN Ling XU Jian-wen*
    2014, 45 (1):  26-30.  doi: 10.3969/j.issn.0529-1356.2014.01.005
    Abstract ( )  

    Objective To observe the NeuroD mRNA expression during rat neural system development and to investigate its possible roles in neural development and differentiation. Methods Total mRNA was abstracted from different parts of brains in different development stages with Trizol kit in rats. The mRNA of NeuroD and β-actin was reversely transcripted and amplificated by semi-quantitative reverse transcription-polymerase chain reaction method (RT-PCR). The quantity of NeuroD and β-actin mRNA expression was detected by gel imaging system. Results NeuroD mRNA existed in E7.5. The mRNA expression of NeuroD and β-actin peaked in E10.5 and E21.5, which average gray values were 20437.88±598.28 and 14482.23±1134.49, respectively. The mRNA expression of NeuroD and β-actin in E10.5 and E21.5 was significantly higher than other time points (P<0.01). The ratios of NeuroD to β-actin in E12.5,E18.5 and E21.5 were 1.59±0.09、1.61±0.07 and 1.70±0.11 respectively, which were higher than other time points(P<0.01). Conclusion The expression of NeuroD mRNA showed an apparent time-specific pattern during the development of the rat brain. The expression level of neuroD increased obviously in two phases. One was in E10.5, when the original rat brain came into being; the other was the anaphase in neural development, when brain form made the perfection, brain tissue stepped into its maturation. Thus, our results suggested the functions of NeuroD: 1. NeuroD influenced the generation and differentiation of nerve cells; 2. NeuroD played its role in the maturation of nervous system.

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    Effects of sericin on Akt signal transduction pathway of hippocampus of the rat with diabetes mellitus
    SONG Cheng-jun ZHANG Yan HE Ya-qiang CHEN Zhi-hong*
    2014, 45 (1):  31-36.  doi: 10.3969/j.issn.0529-1356.2014.01.006
    Abstract ( )  

    Objective To investigate the effects of sericin (obtained by soaking, decocting and concentrating from silkworm cocoon) on Akt signal transduction pathway of hippocampus of the rat with diabetes mellitus. Methods Thirty-six male SD rats were randomly divided into 3 groups, 12 rats per group: normal control group, diabetes mellitus model group and sericin treatment group. The type 2 diabetes mellitus rat model was established by continuous peritoneal injection of streptozotocin with a small dose (25mg/kg/d, 3d). The rats in sericin treatment group were lavaged with sericin for 35d (2.4g/kg·d). TUNEL was used to detect neurons apoptosis in CA1 region of hippocampus; Western blotting and RT-PCR to detect the expression of Akt signal transduction pathway related factors: Akt, NF-κB and Bad. Results Compared with normal control rats, the apoptotic index of hippocampal neurons of diabetes mellitus rats increased obviously (P<0.01); Akt and NF-κB expression in hippocampus decreased obviously, Bad expression increased obviously (P<0.01). Compared with diabetes mellitus rats, the apoptotic index of hippocampal neurons of rats in sericin treatment group decreased significantly (P<0.05); Akt and NF-κB expression in hippocampus increased significantly, Bad expression decreased significantly (P<0.01, P<0.05). Conclusion Sericin can reduce hippocampal neurons apoptosis by regulating abnormal changes of Akt signal transduction pathway, thus sericin has fairly protective effects on hippocampus injury during diabetes mellitus.

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    Effects of remyelination of Olig on oligodendrocyte mediated demyelination in the rat
    YAO Hong-bo ZHANG Meng GONG Xue-wu SUN Li-hui*
    2014, 45 (1):  37-40.  doi: 10.3969/j.issn.0529-1356.2014.01.007
    Abstract ( )  

    Objective To investigate the role of oligodendrocytes mediated demyelination in the rat about Olig in myelin regeneration. Methods Forty healthy Wistar rats were randomly divided into normal group, model control group, model group and research group. Morphological observation and Immuno- histochemistry detection expression of Olig1 and Olig2 were performed. Results Olig1 was located the cytoplasm of oligodendrocytes in normal group, nuclear expression was increased significantly in model group, and Olig1 was retransferred to the cytoplasm in the research group. Expression of Olig2 was located in the nucleus in the normal group. A few of Olig2 were expressed in the cytoplasm in the model group. Conclusion The brain may not be able to generate oligodendrocytes when Oligl and Olig2 are deficient, which is affect of the myelin regeneration.

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    Distribution of monoaminergic neurons in the central nervous system of adult tree shrew
    LAN Wei DING Yu-qiang*
    2014, 45 (1):  41-46.  doi: 10.3969/j.issn.0529-1356.2014.01.008
    Abstract ( )  

    Objective To explore the distribution of tyrosine hydroxylase (TH) and tryptophan hydroxylase 2 (Tph2) in the CNS of the adult tree shrew (Tupaia belangeri chinensis). Methods TH and Tph2 expressions in the adult tree shrew were localized using immunohistochemistry. Results TH-immunoreactive signals were mainly distributed in the glomerular layer of the olfactory bulb, striatum, paraventricular hypothalamic nucleus, subtantia nigra, ventral tegmental area and locus coeruleus. Tph2- immunoreactive signals were mainly located in the midbrain and the pontine raphe complex. Conclusion The distribution pattern of TH and Tph2 in the tree shrew CNS is consistent with that of most vertebrate. Some distribution characteristics shown in the tree shrew support its close afnity to primates. The distribution pattern of TH and Tph2 in the tree shrew CNS indicates the conservation of the evolution of the dopamine and serotonin systems, and suggests that the tree shrew can be a potential animal model to study human mental diseases associated with the dopamine and serotonin systems.

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    Compatibility study of bone mesenchymal stem cells and ferulic acid treatment PHBV film in vitro
    SONG Zhang-shuo BAI Shu-ling* PAN Feng TIAN Xiao-hong TONG Hao
    2014, 45 (1):  47-52.  doi: 10.3969/j.issn.0529-1356.2014.01.009
    Abstract ( )  

    Objective To evaluate the growing condition of bone mesenchymal stem cells (BMSCs) which were cultivated in the film of different concentration in the ferulic acid treatment of abamectin poly-3-hydroxy butyrate-co-3-hydroxy valerate(PHBV), and to investigate biocompatibility between the membrane of poly-3-hydroxy butyrate-co-3-hydroxy valerate and bone mesenchymal stem cells to provide a basis for the choice of scaffold material of bone tissue engineering. Methods The BMSCs were harvested by a whole bone marrow culture method,and identified by flow cytometric to analysis detection cell surface antigen. The BMSCs of the third passage were inoculated on the material surface and cultured for 8 hours,12 hours and 24 hours. Changes in form of BMSCs on different concentrations in the ferulic acid treatment surfaces were observed by scanning electron microscopy(SEM).The material leaching liquor to culture BMSCs was prepared and material was detected with the influence of cells by MTT and flow cytometer cell cycle assays method. Results The BMSCs were successfully cultured, these cells were uniformly negative for CD44,CD90 and positive for CD34,CD11b.SCM showed that PHBV material surface coarse, visible crystals.The cells on the group of a and b material surface growed well,and proliferated significantly.The MTT analysis and flow cytometer cell cycle assays showed that the PHBV film of 5% ferulic acid treatment promoted the growth and proliferation of BMSCsin vitro. Conclusion The PHBV film with 5% ferulic acid treatment had good biocompatibility of BMSCs. Environment is conducive to cell growth, adhesion and proliferation in vitro.

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    Effect of hyperthermia to apoptosis on mice thymocytes
    ZHANG Jin-hong ZHANG Lan WANG Fang*
    2014, 45 (1):  53-57.  doi: 10.3969/j.issn.0529-1356.2014.01.010
    Abstract ( )  

    Objective To study the effect of hyperthermia to apoptosis on mice thymocytes. Methods One-month-old male mice were divided into 6 groups randomly (A-F).5 mice per group, and 1 control group (A) and 5 experimental groups (B-F). Mice in experimental groups were stressed for 2 hours at 37℃ and then raised at room temperature(RT). At the different time points (0hour,2hours,6hours,10hours,20hours) after RT recovering, thymuses from one group were collected. Transmission electron microscopy was used to observe cellular morphology. Flow cytometry and hematoxylineosin (HE) staining were used to analyze apoptosis and cell cycle of thymocytes. Expression of proliferating cell nuclear antigen(PCNA) and heat shock protein(HSP70) were detected by immunohistochemistry. Results Typical morphology of apoptotic cells were observed under transmission electron microscopy at 2hours, 6hours and 10hours after hyperthermia. At 6hours after hyperthermia, thymocytes of S phase and apoptosis reached the max while thymocytes of G0/G1 phase and G2/M phase reduced to the minimum. At the same time point, PCNA expressed most. But HSP70 began to express at 0hours and expressed most at 2-6hours after hyperthermia. Conclusion Apoptosis of thymocytes was obviously effected by hyperthermia. HSP70 had protective effect during apoptosis.

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    Regulation of resveratrol to key enzyme in homocysteine metabolism in insulin resistance model
    FAN Xiao-ming ZHANG Tie-jun KANG Li LIU Shi-chang GUO Jia-zhi LU Di GUI Li* LI Shu-de*
    2014, 45 (1):  58-63.  doi: 10.3969/j.issn.0529-1356.2014.01.011
    Abstract ( )  

    Objective To investigate the regulation mechanism of resveratrol(RES) on key enzymes of homocysteine metabolism for SD rats with insulin resistance.
    Methods Six weeks healthy mice were made as the insulin resistance model with diet by high fat and sugar and gavage by high fat for 3 months. The rats were randomly divided into the insulin resistance group (n=10)and the resveratrol intervention group (n=10). High fat and sugar were fed in the insulin resistance group for eight weeks. The resveratrol intervention group was fed by high fat and sugar and gavage by resveratrol[30mg/(kg·d)] for eight weeks. The concentrations of homocysteine, glucose, insulin, total cholesterol and triglyceride were measured in the fasting state. The insulin resistance index (IRI) was calculated. The expressions of methylenetetrahydrofolate reductase (MTHFR), cystathionine β synthase (CBS) and methionine synthetase (MTR) were detected by immunohistochemical, RT-PCR and Western blotting in the rat liver tissue. Results The concentrations of the blood glucose, blood insulin and triglyceride in the resveratrol intervention group were lower than that in the insulin resistance group (P<0.05). The ISI was lower in the resveratrol intervention group. The concentration of homocysteine was decreased (P<0.05). However, the weight and the concentration of total cholesterol were no statistical significance (P>0.05). The protein expressions of MTHFR and CBS in the resveratrol intervention group were higher compared with the insulin resistance group by Western blotting and immunohistochemical staining(P<0.05).The mRNA expressions of MTHER and CBS in the resveratrol intervention group were higher compared with the insulin resistance group by RT-PCR. However, the expression of MTR protein and mRNA were not different between the resveratrol intervention group and the insulin resistance group (P>0.05). Conclusion Resveratrol can improve the expression of the MTHFR and CBS which participate the metabolism of homocysteine and facilitate the transformation of homocysteine, thus it may play a key role in alleviating insulin resistance.

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    Expressions and clinical significance of ROBO4 in both serum and tumor tissues in patients with epithelial ovarian cancer
    MA Rui-qiong YE Xue CHENG Hong-yan CHEN Jun CUI Heng WEI Li-hui CHANG Xiao-hong*
    2014, 45 (1):  64-69.  doi: 10.3969/j.issn.0529-1356.2014.01.012
    Abstract ( )  

    Objective To explore the levels of ROBO4 and its roles in patients with ovarian tumor, and to investigate the correlation between the expressions of ROBO4 in ovarian tumor tissues and the related clinicopathological characteristics. Methods We examined expression of ROBO4 in 40 cases of the normal ovarian tissue, 54 cases of the ovarian cyst tissue, and 110 cases of ovarian cancer by using the immunohistochemistry and RT-PCR, and compared with clinicopathological features of ovarian cancer. Serums were checked by ELISA in patients with ovarian cancer and ovarian benign tumors and health women.
    Results The intensities of ROBO4 protein immunohistochemical positive staining and its mRNA expression were significantly lower in ovarian cancer than those in both normal ovarian tissues and benign tumor tissues (P<0.05). The serum ROBO4 in patients with malignant ovarian tumor was significantly lower than those in healthy women and benign tumors (P<0.05). Loss or reduced expression of ROBO4 was related with clinical stages, lymph node metastasis or omental metastasis or ascites of ovarian cancer (P<0.05), but not related with age, pathologic types, and histological grade. The correlation analysis showed that there was no correlation between serum ROBO4 and CA125. Conclusion Reduced expression of ROBO4 plays an important role in tumorigenesis of ovarian cancer, which is related to invasion and metastasis of ovarian cancer.

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    Effects of microRNA-100 on mitosis block and Polo-like kinase 1 expression in hepatocellular carcinoma cells
    FAN Bing-lin JI Xiao-hui ZHU Wu-ling*
    2014, 45 (1):  70-73.  doi: 10.3969/j.issn.0529-1356.2014.01.013
    Abstract ( )  

    Objective To investigate the effect of microRNA-100(miR-100) on mitosis block and Polo-likekinase 1(PLK1) expression in hepatocellular carcinoma cells. Methods Expression of miR-100 and PLK1 was determined by quantitative real-time PCR (qRT-PCR) and immunofluorescence (IF) in hepatocellular carcinoma HepG2 cells. HepG2 cells were transiently transfected by miR-100 mimic with Cy3 fluorescence labeled through Oligofectamine liposomes, followed by cell mitosis and PLK1 expression analysis. Results Expression level of miR-100 decreased whereas PLK1 mRNA and protein expression abnormally increased in HepG2 cells compared with the normal hepatocytes. At 48hours after transfection of miR-100 mimic, the mitotic index of HepG2 cells in experimental group was significantly less than that of control cells. Western blotting analysis indicated that PLK1 protein expression levels were significantly reduced in experimental cells compared with the control cells. Immunocytochemical results showed that PLK1 protein in the nucleus almost disappeared in meta/ana-phase and telophase of mitosis. Conclusion MiR-100 may inhibit the expression of PLK1 protein, which leads to mitotic block of hepatocellular carcinoma cells.

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    Effects of TWEAK antisense oligonucleotide on the proliferation and invasive capability of the human colon cancer cell line SW480
    ZHANG Yan-xin* ZHAO Hong-jun SONG Wen-gang GAO Feng-lan Lü Hui-fang
    2014, 45 (1):  74-79.  doi: 10.3969/j.issn.0529-1356.2014.01.014
    Abstract ( )  

    Objective To explore the effects of TNF-like weak inducer of apoptosis(TWEAK) antisense oligonucleotide on the proliferation and invasive capability of the human colon cancer cell line SW480. Methods Colon cancer SW480 cells were transfected with synthesized TWEAK antisense oligodeoxynucleotide (ASODN) and scrambled oligodeoxynucleotide (SCODN). The cells were divided into control group, LF control group, ASODN group and SCODN group; MTT was applied to detect the inhibition of tumor cell proliferation, flow cytometry to examine the cell cycle distribution, ELISA to investigate the expression of soluble TWEAK and its receptor Fn14 in supernatant of cell culture fluid, Western blotting and immunofluorescence cytochemistry(IF) to detect the expression of soluble TWEAK and its receptor Fn14 and Transwell invasion assay to observe the changes on invasive capability of colon cancer cells. Results There was no significant difference on proliferation between the control group, LF control group and SCODN group (P>0.05). Compared with the control group, the proliferation rate of ASODN group tumor cells was inhibited obviously (P<0.05) in a dosage-time dependent manner. The ratio of G2+M phase cells was remarkably higher than that of control group after transfection with TWEAK ASODN (P<0.05). The expression rates of TWEAK and its receptor Fn14 in both cell culture fluid and cells were lower than those of control group, and the difference was of statistical significance (all P<0.05). The inhibition rate of infiltration was 31.39% after transfection with TWEAK ASODN, higher than those of control groups (P<0.05). The expression of TWEAK and its receptor Fn14 in cell culture fluid and cells was closely correlated (all P<0.05). Conclusion TWEAK ASODN may affect the occurrence and development of tumor probably by inhibition of binding between TWEAK and its receptor Fn14, and inhibition of TWEAK protein expression may restrain the proliferation and infiltration of the human colon cancer cell line SW480.

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    Microanatomical and clinical significance of the recurrent laryngeal nerve in the thyroid region
    WANG Qin HU Guo-qin1 XU Sheng-chun*
    2014, 45 (1):  80-83.  doi: 10.3969/j.issn.0529-1356.2014.01.015
    Abstract ( )  

    Objective To investigate the anatomic characteristics of the recurrent laryngeal nerve in the thyroid region. Methods The recurrent laryngeal nerve and its surrounding structures of 48 adult cadavers were examined with gross anatomical technique. Results The relationship between the recurrent laryngeal nerve and the lateral cricopharyngeal triangle in 48 adult cadavers (96 sides) was detected. The main trunk of the recurrent laryngeal nerve coursed superiorly and entered into the lateral cricopharyngeal triangle either through (29%,28/96 sides) or deep to (71%,68/96 sides) the cricopharyngeal muscle. The entering point of recurrent larynageal nerve into the larynx was(5.9±1.8)mm inferior to the inferior cornu of the thyroid cartilage. At the level of(15.2±5.5)mm inferior to the inferior cornu of the thyroid cartilage,69% (66/96 sides) of the recurrent laryngeal nerve gave off a small branch to the larynx and its entering point into the larynx was (4.8±2.1) mm inferior to the inferior cornu of the thyroid cartilage. The spatial relationship between the recurrent larynageal nerve inferior to connective tissue membrane and the inferior thyroid artery branch was below the triangle, and the nerve ran either in front of (66%,63/96 sides) or posterior to (33%,32/96 sides) the inferior thyroid artery. Conclusion Correct identification and safe dissection of the recurrent laryngeal nerve are essential in surgery. Most of the recurrent larynageal nerve have external laryngeal branch. It is necessary to confirm the entering point of recurrent larynageal nerve into the larynx while trying to find recurrent larynageal nerve based on the location of the inferior cornu of the thyroid cartilage. The relationship between the inferior thyroid artery and the RLN is complicated, thus the inferior thyroid artery may not be considered as the first choice to localize the recurrent laryngeal nerve.

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    Endoscopic anatomy of a double nostrils surgical approach to Meckel’s cave
    BAI Zhi-qiang JIN Yong* TAO Bao-hong WU Zhong-min
    2014, 45 (1):  84-88.  doi: 10.3969/j.issn.0529-1356.2014.01.016
    Abstract ( )  

    Objective To study the structures relevant to the endoscopic approach to the Meckel’s cave via the amphirhinal in order to provide the anatomy and morphology information for the Meckel’s cave surgery. Methods By simulating the operative approach to the Meckel’s cave via the amphirhinal under the endoscope, the operative approach-relevant anatomic landmarks in 10 cadaveric heads (20 sides) were observed, analyzed, photographied and measured. The cadavers were arterially and venously perfused with colored latex. Results The approach to the Meckel’s cave via the amphirhinal was divided into 4 steps: finding ostium of the maxillary sinus,entering the maxillary sinus,entering pterygopalatine fossa(PPF)and entering the Meckel’s cave. The distance from the nasal columella to the ostium of the maxillary sinus was (45.07±2.01) mm;the distance from the nasal columella to the sphenopalatine foramen was(64.84±3.00) mm;the distance from the nasal columella to the anterior foramen of the pterygoid canal was (71.34±2.99) mm. The angle between the lines from nasal columella to posterior nasal spine and from nasal columella to apertura maxillaries was (38.81±1.72)°. The angle between the lines from nasal columella to posterior nasal spine and from nasal columella to sphenopalatine foramen was(25.92±2.05) °. The diameter of sphenopalatine artery was (2.21±0.24)mm, and the diameter of vidian artery was (1.07±0.27)mm. The PPF region was structurally complicated due to the inside maxillary artery and its terminal branch-sphenopalatine artery, and great variation of the descending palatine artery. Tracing the sphenopalatine artery made it easier to localize the maxillary artery and its branches. When the constitutions such as pterygopalatine ganglions, veins were carefully departed, the vidian nerve and vessel bundles were tracked. The vidian ending was characterized to face with the ruptured fractions of the internal carotid artery. Meckel’s cave was straightly accessed by anatomically departing the quadrangular space (QS). Conclusion The selection of the operative approach to Meckel’s cave should be individualized. The location and the size of the lesion in Meckel’s cave are the major factors in deciding approaches for surgical treatment. The endoscopic approach of double nostrils-Meckel’s cave provides direct exposure to the anteromedial compartment of Meckel’s cave and PPF. Notable anatomical landmarks for the endoscopic surgeon are the sphenopalatine foramen, the vidian nerve and its canal and maxillarynerve (V2). A dissection superficial to the vascular structures preserves the neural elements. The nerves, such as the vidian nerve and V2, and their bony foramina are critical anatomical landmarks to identify and control the ICA( internal carotid artery) at the skull base.

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    Enoloscopic anatomy of the endonasal transmaxillary transpterygoid approach to Meckel’s cave
    GU Ye ZHANG Xiao-biao*WANG Xue-jianYU Yong HU FanXU Wen-long XIE Tao SUN Chong-jing
    2014, 45 (1):  89-93.  doi: 10.3969/j.issn.0529-1356.2014.01.017
    Abstract ( )  

    Objective To investigate anatomic characteristics of an endoscopic endonasal transmaxillary transpterygoid approach to Meckel’s cave, which included searching significant anatomic landmarks and obtaining relevant anatomic measurements. To establish an anatomic basis for clinical applications of endoscopic endonasal transmaxillary transpterygoid approach to Meckel’s cave. Methods Five adult fresh head specimens (10 sides) were investigated to identify important anatomic landmarks. The distances between the landmarks in the endoscopic endonasal transmaxillary transpterygoid approach to Meckel’s cave were measured during the anatomic procedure. Imaging navigation system (IGS) was employed. Results The distances from columella nasi (NC) to choana, sphenoid ostium (SO), anterior aperture of palatosphenoidal canal (PSC), sphenopalatine foramen (SPF), and the anterior aperture of vidian canal (VC) were(66.5±3.3) mm, (61.2±1.6) mm, (64.6±1.4) mm, (62.8±2.3) mm, and (75.4±3.3) mm, respectively. The distances from anterior aperture of VC to anterior aperture of PSC and foramen rotundum (FR) were (2.1±0.7) mm and (7.5±0.7) mm. The lengths of PSC and VC were (6.4±0.5) mm and (13.3±1.2) mm. The anterior aperture of VC was identified via finding PSC. VC was considered a landmark to the anterior genu of petrous internal carotid (ICA). Paraclival ICA was be identified by paraclival carotid prominence (CP). Imaging navigation system (IGS) helped identifying anatomical landmarks and guiding anatomic manipulating. Conclusion Fully exposure of Meckel’s cave via the endoscopic endonasal transmaxillary transpterygoid approach is feasible. The approach enters into anterior Meckel’s cave through a quadrangular space, which is bordered medially and inferiorly by ICA, laterally by maxillary nerve, and superiorly by the abducens cranial nerve. The data from the experiment and IGS is useful to locate crucial anatomical landmarks.

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    Anatomical measurements of the structures relevant to the lumbar spondylolysis articular process and isthmus
    RUAN Cai-lian*
    2014, 45 (1):  94-97.  doi: 10.3969/j.issn.0529-1356.2014.01.018
    Abstract ( )  

    Objective To measure the structures relevant to the spondylolysis process and isthmus of the lower lumbar spine by using the imaging method, and to analyze the new lumbar posterior planting bone surgery operation in the process of the safety range and the patients with the relationship between the height and weight. Methods Lateral lumbar spine X-ray images were collected from 300 normal adult males and females, 150 each. The width, length, depth, the spondylolysis depth, graft length and absolute depth of the superior articular process of the 3rd-5th lumbar vertebral were measured with the Digimizer software. The equation of linear regression method was used to analyze these indicators and the patients’height and weight. Results The width, length, depth and the depth of spondylolysis and absolute depth of the articular process, and patients’weight and height in both males and females were not correlated (P> 0.05). There was a significant correlation between planting bone bed length and height (P< 0.05), but not the weight (P> 0.05). Conclusion The image method measuring the lower lumbar spine on the joints and outstanding spondylolysis anatomical structure related to posterior lumbar for bone graft operation provides accurate and reliable operation data.

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    Cardiac microvascular endothelial cell injury induced by microwave irradiation and endoplasmic reticulum stress
    QU Xiao-ling LI Yu-zhen LIU Xiu-hua SONG Zhi-yuan*
    2014, 45 (1):  98-103.  doi: 10.3969/j.issn.0529-1356.2014.01.019
    Abstract ( )  

    Objective To study the relationship between the damage of cardiac microvascular endothelial cells by microwave radiation and endoplasmic reticulum stress. Methods Cardiac microvascular endothelial cells of the rat that were cultured 3-4 generation were divided into control and radiation groups. Of radiation groups, using 10mW/cm2,30mW/cm2,50mW/cm2 microwave radiated cardiac microvascular endothelial cells, respectively, and radiated for 6min. After radiated 24hours,cells were collected. The cells were exposed to 30mW/cm2 microwave for 6 minutes. After cultured for 1hour, 3hours or 24hours, endothelial cells were collected and the control group finished experiment at 24 hours. The annexin V-propidium iodide double staining method was used to detect apoptosis rate. The method of phalloidin staining to observe the changes of microvascular endothelial cytoskeleton was used. Westen blotting was used for detecting the protein expression of calreticulin, CHOP and GRP78. Results The doseeffect study on the apoptosis showed that, after microwave irradiation, the apoptosis rates in 10mW/cm2, 30mW/cm2, 50mW/cm2 irradiation groups were (2.34±0.15)%, (2.72±0.96)%, (2.62±0.34)% and there were significant difference (P<0.05) comparing to the control group(0.88±0.32)%. Aging studies on the apoptosis found that, at 1hour, 3hours and 24hours of post exposure to 30mW/cm2,the apoptosis rate of cells were (1.12±0.15)%, (1.49±0.54)% and (1.85±0.45)%.The control group was (1.10±0.28)%.Post irradiated 1hour group had no significant difference (P>0.05), post irradiated 3hours group and 24hours group had significant difference (P<0.05). Molecular detection of endoplasmic reticulum stress showed that, comparing with the control group, the protein expressions of CRT, GRP78 and CHOP in the 30mW group increased by 124%, 76% and 256%, respectively. In the 50mW group,the protein expressions of GRP78 and CHOP increased by 52% and 189%, respectively. Comparing with the control group, the protein expressions of CRT, GRP78 and CHOP in the 30mW and GRP78 and CHOP in the 50mW group showed significant differences (P<0.05). The protein expressions of CRT, GRP78 and CHOP in the 10mW and CRT in the 50mW group showed no significant differences (P>0.05). Cytoskeleton staining showed that, under the control conditions, endothelial cells displayed a few actin stress fibers. Exposure of endothelial cells to microwave caused a dramatic increase in the number of F-actin stress fibers. Maximal stress fiber formation occurred when endothelial cells were challenged for 3h or with 30mW microwave. Conclusion Microwave radiation can induce serious endoplasmic reticulum stress, resulting in cardiac microvascular endothelial cell damage.

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    Effect of testosterone on the in vitro mouse follicle development
    MU Xiao-li LIU Tao LIU Hong-bin GAO Yuan GAO Xuan GENG Ling YOU Li XU Cheng-yan*
    2014, 45 (1):  104-108.  doi: 10.3969/j.issn.0529-1356.2014.01.020
    Abstract ( )  

    Objective To investigate the effect of testosterone on the in vitro mouse follicle development. Methods We chose follicles of 12-day-old ICR female mice culturedin vitro as our research models. Follicles were divided into four groups randomly with thirty follicles in each group and were treated with different concentration testosterone: 0(control group), 10-4 mol/L,10-5 mol/L,10-6 mol/L. Follicles were divided into three groups randomly with thirty follicles in each group and were treated with different reagents: no treatment (control group), 10-5 mol/L testosterone(group T), 10-5 mol/L androgen antagonist flutamide pretreatment plus 10-5 mol/L testosterone(group F+T). Follicles were cultured on microdrop. We observed follicle outlines, granulosa cells proliferation and oocytes morphology under the inverted microscope every day, recorded follicle degradation. Results The development of follicles was inhibited in the group treated with 10-4 mol/L testosterone. While the granulosa cells proliferation and the development of follicles were promoted in the group treated with 10-5 mol/L testosterone. Meanwhile, testosterone stimulated the follicles development in the early stage of the follicular development, but it inhibited the follicles development in the late stage. Conclusion 10-5 mol/L testosterone can promote the development of primary follicles and secondary follicles, but it can inhibit the antral follicles development.

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    Smad3 promotes the autophagy of granulosa cells in rats
    SHEN Chun-yan DONG Jing-xia XU Jian*
    2014, 45 (1):  109-113.  doi: 10.3969/j.issn.0529-1356.2014.01.021
    Abstract ( )  

    Objective To study the effect of Smad3 on the autophagy of the ovarian granulosa cells (GCs) in the rat. Methods Ovarian granulosa cells were cultured and treated in the following conditions: control group in which GCs were cultured without any treatment; knockdown group in which GCs were transfected by specific Smad3 siRNA with Lipo RNAiMAX; overexpression group in which GCs were transfected by p3×FLAG-CMV-Smad3 plasmid with Lipo 2000. Immunocytochemistry was used to identify cell purity. To determine the efficiency of transfection, the expression of Smad3 protein was evaluated by Western blotting; Western blotting analysis was also used to detect the expression of autophagy associated gene 8(LC3), B cell lymphoma2(Bcl-2) proteins of ovarian GCs. Results Western blotting results showed that: in knockdown group, the ratio of LC3B II/I had no significant difference and the expression intensity of Bcl-2 protein was significantly decreased, compared to the control group. In overexpression group, the ratio of LC3BII/I was significantly increased and the expression intensity of Bcl-2 protein was significantly increased, compared to the control group. Conclusion Both Smad 3 SiRNA and P3×FLA-CMV-Smad 3 can be effectively transfected to GCs. The mechanism, by which Smad 3 promotes the autophagy of GCs, may be related to the expression of Bcl-2 protein.

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    Effect of eucalyptus oil on rat embryo implantation and growth interfered with retinoic acid
    SU Li-li HE Ying-hong CHEN Mo YU Yong-li* HUANG Yi-ling ZHU Cheng-xue LIU Mao-sheng
    2014, 45 (1):  114-119.  doi: 10.3969/j.issn.0529-1356.2014.01.022
    Abstract ( )  

    Objective To study the effect of eucalyptus oil on retinoic acid(RA) interference in rat embryo implantation and growth. Methods Forty-two pregnant SD rats were randomly divided into 6 groups,7 rats each: 3 experiment groups ( high, medium, low doses of Eucalyptus+RA), a solvent group (peanut oil +RA), a retinoic acid group (RA) and a normal group with normal diet. Three experiment groups and Solvent group were lavaged from gestation 7th to 14th day with eucalyptus oil in 300 mg/kg, 200 mg/kg, and 100 mg/kg and peanut oil 2ml per animal per day, respectively. Each of them was garaged with retinoic acid (40mg/kg) at gestation 10th day. RA group was garaged with 40 mg/kg at gestation 10th day. All pregnant rats were sacrificed at 21st day. The weights of the body, uterus and placenta of the pregnant rats were measured. The number of absorbed, live and dead fetus were counted respectively. The length of fetus body and tail as well as body weight were measured. Results The uterus and placenta weight, change in body weight of pregnant rats in normal group, Eucalyptus oil groups and Solvent group showed no significant difference compared with RA group. The solvent group rats’ placental weight was lower than that in eucalyptus oil groups’ and RA group’s (P>0.05), but it was significantly lower than that in normal group (P<0.05). The rate of live fetus in normal group was more than that in other groups with significant difference (P<0.05). The teratogenic rate and the absorption rate of normal group were lower than that of the other groups’ (P<0.05). The mortality of normal group was zero. In gavaged retinoic acid groups, the rates of live fetus in high, medium of eucalyptus oil group were higher than that in RA group (P>0.05). It’s teratogenic rate was lower than that of RA group(P>0.05). The absorption rate in each eucalyptus oil group and solvent group compared with RA group had no difference. The fetus body weight, body length, and tail length of Normal group were significantly higher than that in the other groups(P<0.05). In gavaged retinoic acid groups, the fetus body weight and body length of each eucalyptus oil group were significantly higher than that in solvent group and RA group(P<0.05). The fetus body weight, body length, tail length of solvent group had no significant difference compared with the RA group (P>0.05). The fetus tail length in eucalyptus oil groups was higher than that in RA group, but no significant differences were found(P>0.05). Conclusion Retinoic acid interferes the rat embryo implantation and development; Eucalyptus oil has certain influence on the RA interference of rat embryo implantation, and it also has certain inhibition on fetal growth retardation.

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    Light and transmission electron microscopic observation on the gill sturcture of juvenile Pseudosciaena crocea
    RUAN Cheng-xu WU De-feng* YUAN Chong-gui
    2014, 45 (1):  120-123.  doi: 10.3969/j.issn.0529-1356.2014.01.023
    Abstract ( )  

    Objective  To study the microscopic structure and ultrastructure of gill of juvenile Pseudosciaena croceafor more abundant reference of breathing and salinity adaptation physiology. Methods The microscopic structure and ultrastructure of gill of juvenilePseudosciaena crocea were observed by light and transmission electron microscopic. Results The structures of gill of juvenilePseudosciaena crocea were similar to those of other teleosts, the comb primary filaments lined on the gill, and secondary gill lamellae inlaid on two sides of each filament. Secondary filaments consisted of epithelial cells, endothelial cells, pillar cells and capillary vessel net. Chloride cells were found both in the secondary filaments and the base. In the base of the secondary filaments, the number of chloride cells were more. Inside the cells there were tubular network and abundant mitochondrias. Conclusion The structures of gill of juvenilePseudosciaena crocea were similar to those of other teleosts. The features of the structure of gill maybe can explain whyPseudosciaena croceahave strong adaptability to salinity.

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    Digital patterns in patients with coronary heart disease
    LU Hong JIA Fei ZHU Hao DANG Jie ZHAO Jun-li HUO Zheng-hao*
    2014, 45 (1):  124-127.  doi: 10.3969/j.issn.0529-1356.2014.01.024
    Abstract ( )  

    Objective To investigate the relationship between digital patterns and the coronary heart disease. Methods Using anthropometry, the frequency of finger print patterns of 272 men from Ningxia Han nationality that consisted of 136 patients with coronary heart disease and 136 normal adults was studied. The differences of them were analyzed. Results Patients with coronary heart disease had higher digital frequencies of whorls in both hands as compared with controls (P<0.01). The descending order of both group was Ⅳ> Ⅰ> Ⅱ>Ⅲ>Ⅴ; Frequency of more than 6 digital whorls in the coronary heart disease group was significantly higher than that of the controls (P<0.05). Conclusion Frequency of digital whorls (W) may be one of the important early diagnostic index of coronary heart disease.

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    LIN28B gene polymorphisms in Ningxia Hui and Han ethnic people
    PEI Li-guo QU Lei JIA Fei ZHANG Chuan GUO Meng-jing LU Hong HUO Zheng-hao*
    2014, 45 (1):  128-133.  doi: 10.3969/j.issn.0529-1356.2014.01.025
    Abstract ( )  

    Objective To study the frequencies of rrs314277 and rs314276 allele and genotype distribution of the LIN28B gene polymorphisms in the Hui and Han people in Ningxia, and to analyze the distributions of LIN28B gene polymorphisms among different places and ethnicity. Methods A total of 705 subjects were from Ningxia Medical University, including 311 cases of boys, of which 144 cases of the Hui and 167 cases of the Han; 394 cases of girls, including the 188 cases of Hui and the 206 cases of Han were analyzed in this study. The two SNPs were examined by the sequencing methods. Results The genotypes frequencies and allele gene frequency of LIN28B gene in Ningxia Hui population were: male:rs314277 (AC:4.5%,CC:38.9%; A:2.3%, C:41.1%); rs314276 (AA:2.7%,AC:21.4%,CC:19.3%; A:13.4%,C:30.0%);female:rs314277(AA: 0.6%, AC:7.5%,CC:48.5%; A:4.4%, C:52.3%); rs314276(AA:3.3%,AC:28.9%, CC:24.4%; A:17.8%,C:38.9%). The genotypes frequencies and allele gene frequency of LIN28B gene in Ningxia Han population were: male:rs314277(AC:3.2%, CC:41.6%; A:1.6%, C:43.2%); rs314276(AA: 4.6%,AC:17.4%, CC:22.8%; A:13.3%, C:31.5%); female:rs314277(AC:4.6%,CC:50.7%; A:2.3%, C:52.9%); rs314276(AA: 4.8%, AC:18.8%, CC:31.6%; A:14.2%, C:41.0%). Conclusion This study showed a significant difference of allele between the Hui and Han ethnic of Ningxia population in rs314277(P<0.05), a significant difference of genotypes between the Hui and Han ethnic of Ningxia population in rs314276(P<0.05). No significant differentce is found between the male and female of Ningxia population in the two SNPs (P>0.05). The differences of distribution traits of genotype and allele frequencies between Ningxia population and other different etbnic population in LIN28B gene indicate that this two SNPs may have genetics characteristic in different ethnics.

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    Circumference and its variation with age of rural adults of Hakka in Jiangxi
    ZHENG Ming-xia ZHENG Lian-bin* LU Shun-hua BAO Jin-ping YANG Qian ZHOU Hao-le
    2014, 45 (1):  134-139.  doi: 10.3969/j.issn.0529-1356.2014.01.026
    Abstract ( )  

    Objective To study the circumference and its variation with age of rural adults of Jiangxi Hakka group. Methods Using random sampling method to measure the circumference values of 379 adults (183 males,196 females) of Jiangxi Hakka group,including the circumference of head, neck, chest, chest circumference at inspiration,chest circumference at expiration,abdominal,hip,thigh,calf,biceps,forearm,and maximum biceps.The change pattern of circumference values of different age groups was analyzed. Comparison of the circumference values in 18 ethnic group in China was drawn by cluster analysis. Results 1. The result of variance analysis showed that the male and female circumference values of Jiangxi Hakka had significant variation in vrious age groups. 2. A linear correlation analysis showed that with the increase of age,the male chest circumference,chest circumference at inspiration,chest circumference at expiration,abdominal circumference values had a linear regularity increase.Head and calf circumference values had a linear regularity decrease.Female chest circumference,chest circumference at inspiration,chest circumference at expiration,abdominal circumference,biceps circumference, maximum biceps circumference values had a linear regularity increases.Head and thigh circumference values had a linear regularity decrease.3. Hip circumference value had not significant differences between sexes.The other 11 circumference values had statistically significant difference between sexes,the male was significantly higher than the female. Conclusion Circumference values of Jiangxi Hakka group of rural adults have the features of north type groups of our country.

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    Progress in the studies of neural stem cell differentiation
    XING Hua-jie LI Meng-tao SONG Qing BAI Yun*
    2014, 45 (1):  140-144.  doi: 10.3969/j.issn.0529-1356.2014.01.027
    Abstract ( )  

    Neural stem cells (NSCs) exist widely in human and mammalian central nervous system. NSCs are multipotent stem cells and have the ability of self-renewal. They can generate neurons, astrocytes and oligodendrocytes. Since Reynold et al firstly isolated NSCs from the corpus striatum of mice, great progress as in this field has been made. However, the number of NSCs in central nervous system (CNS) is relatively small. And as NSCs transplanted into animal brain they mainly differentiate into glial cells and form cicatrix tissue, which restricts the repair of CNS. These factors affect the applications of NSCs in clinical medicine. So how to achieve the directed differentiation is a crucial tissue in this field. The research progress is reviewed in this paper.

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