Table of Content

2018, Volume 49 Issue 3
06 June 2018 Previous Issue    Next Issue
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Neurobiology
Histology,Embryology and Developmental Biology
Anatomy
Cell and Molecules Biology
Cancer Biology
Technology and Methodology
Neurobiology
Anatomy
Neurobiology
Cancer Biology
Anthropology
Review


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Neurobiology

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Neural immune change in depression model mice WAN Ren-ling PAN Qi AN Xiao-hong QIAO Cai-li WU Hua-li SHANG Jing 2018, 49 (3):  281-287.  doi: 10.16098/j.issn.0529-1356.2018.03.002 Abstract ( )   PDF (631KB) ( )   Objective To explore the difference of chronic restrain stress (CRS) and chronic unpredictable mild stress (CUMS) model in the neuronal/immune system in 45 male C57BL/6 mice through evaluating behavioral alteration, the hippocampus and the spleen structure and related factor expressions，so as to provide certain references for the pathogenesis of depression and early drug screening for antidepressants. Methods Totally 45 mice were randomly divided into control group，CRS group，and CUMS group．After 6 weeks of stress exposure, Behavioral assessment experiments were carried out to detect the behavior changes of mice. HE staining was to examine structural changes in hippocampus and spleen; The Nissl staining was used to explore neuron damage in hippocampus; In addition, the Real-time PCR was used to detect the gene expression of brain derived neurotrophic factor(BDNF),5-hydroxytrypamine transporter(5-HTT), indoleamine 2, 3 dioxygenase 1（IDO1）in hippocampus and inflammation factor IL-1β, IL-6 in spleen. Results Compared with the control group, CRS group had no significant difference in the number of crossing and rearing. However, it was contrary to the CUMS groups; The immobility of tail suspension and forced swimming test were significant increased in both groups; Both stress had a similar damage effect on hippocampus and spleen structure, while CUMS group of hippocampus neuron damage was more serious; Only CUMS group had significant change in the level of gene expression decreased in BDNF and 5-HTT and elevated in IDO1 expression levels; The spleen gene expression levels of IL-1β and IL-6 were significantly increased in CUMS group, while the spleen gene expression levels of IL-6 had no significant change in CRS group. Conclusion The results show that both CRS and CUMS can induce depressive-like behaviors in C57BL/6 mice. From the perspective of neuro-immunology observation, depression symptoms in CUMS model tend to be more obvious. This suggestes that CUMS model may be a better model of depression. References | Related Articles | Metrics

Histology,Embryology and Developmental Biology

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Gill injury by copper and its mechanism in zebrafish ZHAO Qiao-ya SUN Xue-jing WANG Ling-ling WANG Tao-zhi CHEN Qiu-sheng LIN Jin-xing 2018, 49 (3):  367-373.  doi: 10.16098/j.issn.0529-1356.2018.03.016 Abstract ( )   PDF (1324KB) ( )   Objective To study the effect of copper on microstructures and ultrastructures of gill of zebrafish, antioxidant defense system of superoxide dismutase (SOD) activity, malondialdehyde (MDA) content and related immune factors ［tumor necrosis factor alpha (TNF-alpha), interleukin（IL）-6, IL-1 beta) ］in the acute toxicity test.Methods Three copper exposures (0.05 mg/L, 0.1 mg/L, 0.2 mg/L) and a control group were set in this study. Results The copper exposure groups showed varying degrees of damage, such as the epithelial cell swelling, shedding, surface mucus, basal cell hyperplasia and gill lamellae fusion under the light microscope. The mitochondria and rough endoplasmic reticulum cell were swelled, and mitochondrial cristae collapse or the disappearance of oncosis morphology were observed under transmission electron microscope(TEM). SOD activity and MDA content extended upward as increase of the copper ion concentration and exposure time. In addition, copper ion exposure on zebrafish SOD, MDA as compared with the control, showed an induction effect. Compared to the control group, TNF-alpha, IL-6, and IL-1 beta levels increased by different extents after treatment with different concentrations of copper and time. Conclusion The heavy metal copper has obvious damage and immune toxicity to the gill structure and antioxidant system of zebrafish. References | Related Articles | Metrics

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Air-induced uterine endometrial decidualization in mice LIU Ruo-nan MIAO Yan-ping WEI Yan-hui ZHOU Rong-yan LI Xiang-yun 2018, 49 (3):  374-378.  doi: 10.16098/j.issn.0529-1356.2018.03.017 Abstract ( )   PDF (420KB) ( )   Objective To establish a nonsurgical induced decidualization technique in mice, and to explore the air-induced decidua in this study. Methods Clean air (2 μl) was transferred into unilateral uterine horn of 30 pseudopregnant 3.5 days mice by a non-surgical transfer method. The morphology, weight, diameter, cellular differentiation, and decidua-related marker genes in airinduced decidua were detected. Results After air transfer four days, twenty-five pseudopregnant mice displayed different degree of decidualization phenomenon. Compared to the control side uterine horn, air-transferred uterine horns appeared significantly dilation and overweight with increased diameter. The permeability of uterine endometria was extensively spread along the whole horns. Three decidualization-realated marker genes ［cyclooxygenase-2(COX-2),prolactin(Prl)3c1 and Prl8a2］ had a higher expression level. In addition, polykaryocytes was observed in air-induced decidua. Conclusion Mouse uterine endometrial decidualization can be effectively induced by a non-surgical air transfer method . References | Related Articles | Metrics

Anatomy

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High-frequency ultrasound imaging of the median nerve and its clinical significance LI Wen-juan WANG Xia LIU Chun-hong 2018, 49 (3):  361-366.  doi: 10.16098/j.issn.0529-1356.2018.03.015 Abstract ( )   PDF (395KB) ( )   Objective To depict the high-frequency ultrasound imaging of the median nerve of the upper limb in the normal living subject,master the median nerve scanning technique for the measurement of its cross-sectional area (CSA) at 5 easily-identified anatomical sites, reveal its relationship with surrounding tissues,and provide the reference range for clinical diagnosis of peripheral nerve diseases. Methods In 240 healthy volunteers, the CSA of the median nerve were measured by using high-frequency ultrasound at 5 sites which were the carpal tunnel ［rasceta］, forearm midpoint, the point where the nerve passed through the pronator teres, supracondylar humerus and at the midpoint of the humerus. At each site, the measurement was repeated 3 times. The average values of the measurements were analyzed for the correlation between the CSA and height and weight. Results High frequency ultrasound showed that the normal median nerve cross section appeared as a reticular low echo image in a round, oval or triangular shape, whereas its longitudinal section appeared as several parallel low echo bundles with intermittent banded echo segmentation.The CSA of the median nerve at these 5 loci were (10.67±0.94) mm2, (10.25±0.61) mm2, (9.83±0.50) mm2, (11.44±0.43) mm2, and (11.24±0.56) mm2, respectively. There was statistical significance of the CSAs of the median nerve between humeral supracondylar humerus and the midpoint of the humerus and between men and women. Conclusion The display rate of the median nerve in the high-frequency ultrasound probe and the whole process is 100%. There are differences of the CSA in different parts of the median nerve. The CSA of the median nerve is thick in the upper arm and thin in the carpal tunnel and at the forearm. References | Related Articles | Metrics

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Anatomy of anconeus free muscle flap for restoration of thumb oppositional function  GAO Wen-jing SHI Zi-yu HAN Yi-kai SUN Xuan-feng ZHANG Zheng XU Gao-lei ZHANG Zhen-hua 2018, 49 (3):  355-360.  doi: 10.16098/j.issn.0529-1356.2018.03.014 Abstract ( )   PDF (806KB) ( )   Objective To compare the anatomical differences between the anconeus and the abductor pollicis brevis (APB), in order to provide anatomical features of the anconeus for its suitability as the free muscle flap for restoration of thumb oppositional function. Methods Nine Chinese cadavers with elbow, forethigh and plam of hand preserved well were collected. By using standard calipers and Image J 1.45 d, we gave a quantitative description of muscle architecture and the neurovasculature involved in the operative planning of restoration thumb oppositional function in which anconeus free muscle flap is used. Results The mean measures of the anconeus were larger than those of the APB ［anconeus/APB fiber length=(82.0±12.0)mm/(51.6±8.3)mm, area=(936.7±221.8)mm2/(704.4±244.2)mm2］. There was no significant difference between mean fiber angles (anconeus/ APB=60.9°±10.2°/71.2°±11.7°; P>0.05) and neurovasculature ［anconeus/APB:artery diameter=(1.9±0.2)/(2.0±0.5)mm, nerve diameter=(1.7±0.3)/(2.1±0.4)mm; P>0.05］. The length (32.0±3.1)mm and caliber ［diameter=(1.16±0.28)mm］of the vascular pedicle of the anconeus(recurrent posterior interosseous artery) and its venae comitans (diameter=0.8 mm) were sufficient for microsurgical anastomosis. Conclusion The anatomic rationale of the anconeus free muscle flap for thenar reconstruction is suitable and compared to other free muscle flap. It has some advantages: reliable and matching anatomy, minimal donor site morbidity, and the potential to restore a greater degree of opposition and thus function in a one-stage procedure. References | Related Articles | Metrics

Cell and Molecules Biology

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Construction of 4T1 CXCR4 knockout gene stable strain using CRISPR/Cas9 gene editing system LIU Si-nian LUAN Jing-yang ZHANG Yan CAO Guan-hua CAO Guang-jin LI Ling 2018, 49 (3):  303-308.  doi: 10.16098/j.issn.0529-1356.2018.03.006 Abstract ( )   PDF (486KB) ( )   Objective To knock out CXCR4 gene in 4T1 cell by CRISPR/Cas9 system and to construct 4T1 cell strain with knocking out CXCR4 gene stably. Methods Two pairs of sgRNAs that could specifically identify the exons of CXCR4 gene were designed to construct LentiCRISPRv2-sgRNA recombinant plasmid and transformed into competent Stbl3.Then the recombinant plasmids were screened for sequencing and transfected into 293T cell to packaging into lentivirus. After infection of 4T1 cells with lentivirus, stable transfected cells were selected by puromycin,and monoclonal cells were isolated and cultured by limiting dilution method . The genomic DNA of monoclonal cells was extracted and sequenced. The expression of CXCR4 mRNA was detected by Real-time PCR. The expression of CXCR4 protein was detected by Western blotting. Results The LentiCRISPRv2-sgRNA recombinant plasmid was successfully constructed, and a cell strain of knocking out CXCR4 deletion 27 bp was obtained. The expression level of CXCR4 mRNA was reduced, and there was almost no expression of CXCR4 protein in cell strain. Conclusion Recombinant plasmids targeting the CXCR4 gene was obtained by CRISPR/Cas9 gene edit system, and the cell strain with stable knockout CXCR4 gene was successfully screened out. References | Related Articles | Metrics

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Multiple phenotypes in cardiac fibroblasts CHANG Yu-qiao LI Ci-xia JIA Yang-yang GUO Zhi-kun 2018, 49 (3):  317-323.  doi: 10.16098/j.issn.0529-1356.2018.03.008 Abstract ( )   PDF (960KB) ( )   Objective To explore the multiple phenotypes in cardiac fibroblasts (CFs). Methods CFs of neonatal rats were isolated from twelve 1-3-day neonatal SD rats and cultured using combined type I collagenase(w/v, 0.1%) trypsin (w/v, 0.25%) digestion method . Morphological characteristics were observed between primary and subcultured CFs. Common molecular markers for fibroblasts such as vimentin, fibroblast specific protein(FSP1) and discoidin domain receptors 2(DDR2) were detected by immunohistochemical staining. Nanog, c-kit, sca-1, CD73 and CD90 recognized as stem cell markers were evaluated in CFs by immunofluorescence staining, and co-expression of those markers was investigated by three-color immunofluorescence technology. Results By the combined enzyme digestion method , the isolated fibroblasts had large volume. Passage 3 cells exhibited good biological activity, fast proliferation and high purity. Vimentin, FSP1 and DDR2 were all expressed and DDR2 was strongly expressed in CFs. Nanog, c-kit, sca-1, CD73 and CD90 were detected in some CFs. Interestingly, CD73, CD90 and sca-1 were co-expressed with nanog positive CFs respectively and the result showed the positive expression rate of nanog+ /CD73 + was the highest than other three groups (59.02%±8.39%). It had significant different (P<0.01). while the positive rate of nanog+ /CD90+ in CFs had no different with the rate of nanog+ /sca-1+ (P>0.05). And the rate of nanog + /CD90+ and nanog+ /sca-1+had extremely significant different with the rate of CD90+ /sca-1+ (P<0.01). But the positive rate of CD90+ /sca-1+ was significantly lower than that in the other three groups (P<0.01). Conclusion CFs have several subpopulations with stem cell characteristics and multiple phenotype. References | Related Articles | Metrics

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Comparaison of three methods on the effect of myogenic differentiation of porcine dedifferentiated adipocytes HU Xiao-li SUI Li-hong LI Fang-zheng HUAN Yan-jun JIANG Zhong-ling JIANG Nan SONG Xue-xiong 2018, 49 (3):  324-329.  doi: 10.16098/j.issn.0529-1356.2018.03.009 Abstract ( )   PDF (879KB) ( )   Objective To compare the three methods on the effect of myogenic differentiation of porcine dedifferentiated adipocytes (DA), in order to screen the better method for inducing myogenic differentiation of porcine DA. Methods The glucocorticoid, galectin-1 and 5-azacytidine(5-aza) were used to induce myogenic differentiation of porcine DA for 6 days, 15 days and 21 days respectively, and then the morphology of cell differentiation was observed by inverted microscope respectively. After 21 days of induction, the expression of muscle specific protein which desmin and myosin heavy chain(MyHC) was detected by indirect immunofluorescence, and the expression of myogenin(MyoG) mRNA was detected by Real-time PCR. Each method was used for detecting induction effect in 3 groups and repeat three times in each group. Results The morphological observation of induced cell showed that the best myotube morphology and the largest number of multinucleated cells was found via galectin-1 induced, followed by 5-aza, however the higher percent of cell death was occurred during 5-aza induction. The irregular cell morphology, multiform radial protrusion, and some cells of adipogenic differentiation were observed by glucocorticoid induction. The indirect immunofluorescence showed that the strong positive of Desmin and MyHC as well as a high expression of MyoG mRNA was observed in induced cells via galectin-1 method and glucocorticoid method, while the weak positive expression of these two kinds of protein with a low MyoG mRNA level was found in induced cells by 5-aza induction. Conclusion Judged from myotube morphology, differentiation of purity and viability, specific protein and gene expression, we suggest that galectin-1 method is superior to the glucocorticoid and 5-aza method for inducing myogenic differentiation of porcine DA. References | Related Articles | Metrics

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Establishment of doxorubicin induced cardiomyocyte injury model by human induced pluripotent stem cell-derived cardiomyocytes CUI Ning KE Bing-bing WU Fu-jian BAI Rui LIU Tao-yan LI Lei LAN Feng CUI Ming 2018, 49 (3):  309-316.  doi: 10.16098/j.issn.0529-1356.2018.03.007 Abstract ( )   PDF (1164KB) ( )   Objective To establish a humanized doxorubicin-induced cardiomyocyte injury model by using human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Methods The human induced pluripotent stem cells were induced to differentiate into hiPSC-CMs. Then the phenotypes of hiPSC-CMs such including cell viability, calcium transients, oxidative stress and DNA damage were detected after treated with various concentrations of doxorubicin for 24 hours. Results Doxorubicin induced a decrease in cell viability of hiPSC-CMs, destroyed calcium transients, increasing oxidative stress, leading to decrease of mitochondrial membrane potential and causing DNA damage, meanwhile dexrazoxane had cardioprotective effects on hiPSC-CMs. Conclusion Doxorubicin induced cardiomyocyte injury model is successfully established with hiPSC-CMs, overcoming the limitation of difficult access to human cardiomyocytes and the different reaction to drug between species, so could to better study on the mechanism of doxorubicin-induced cardiotoxicity and drug screening. References | Related Articles | Metrics

Cancer Biology

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Enhancing effect of gambogic acid on the sensibility of human gastric carcinoma SGC7901/DDP cells to cisplatin TONG Lei WANG Li-jun YUAN Lei 2018, 49 (3):  337-341.  doi: 10.16098/j.issn.0529-1356.2018.03.011 Abstract ( )   PDF (338KB) ( )   Objective The aim of this study is to investigate the effect of gambogic acid (GA) on the sensibility of human gastric carcinoma SGC7901/DDP cells to cisplatin (DDP) and its possible mechanism. Methods The drug-resistant SGC7901/DDP cells were established by stepwise exposure to DDP. CCK-8 assay was employed to detect the cytotoxic effect of GA and DDP on SGC7901/DDP cells, and the combined effect of GA and DDP was analyzed by Chou-Talalay method . The cell apoptosis was studied by flow cytometry. Western blotting was performed to detect the protein levels of Bcl-2, Bax, Survivin, multidrug resistance-associated protein 2 (MRP2), phosphorylated c-Jun N-terminal kinase (p-JNK) (Thr183/Tyr185) and JNK. Results The IC50 values of GA and DDP were 2.94 μmol/L and 39.76 μmol/L after 48 hours treatment respectively, and the combined both drugs improved the antitumor effects on SGC7901/DDP cells. GA enhanced cisplatin-induced apoptosis (P<0.05), down-regulated Survivin and MRP2 and up-regulated Bax at the protein level (P<0.05), and inhibited phosphorylation of JNK in SGC7901/DDP cells (P<0.05). SP600125, a specific inhibitor of JNK, declined the protein level of MRP2 (P<0.05). Conclusion GA may enhance the sensibility of SGC7901/DDP cells to cisplatin by down-regulating MRP2 as a result of inactivation of the JNK signaling pathway, and up-regulating Bax and down-regulating Survivin. References | Related Articles | Metrics

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Inhibitory effect of dihydromyricetin combined with etoposide on the choriocarcinoma cell JAR WANG Kang XU Qian ZUO Yan-zhen LIU Lei LU Yan-jie LEI Yun-tao CUI Ying LI Yu-hong 2018, 49 (3):  330-336.  doi: 10.16098/j.issn.0529-1356.2018.03.010 Abstract ( )   PDF (696KB) ( )   Objective To investigate the inhibitory effect and related mechanism of dihydromyricetin(DMY) combined with etoposide(VP16) on the choriocarcinoma cell JAR.Methods JAR cells cultured in vitro were devided into control group, DMY group, VP16 group and DMY+VP16 group. The cell proliferation was measured by MTT assay. The apoptosis rate of each group was analyzed by annexin V-FITC/PI double dye flow cytometry. The cell viability was measured by the colony formation capability assay. The protein expression levels of Bcl-2, inhibitor apoptosis protein 2(c-IAP2) were detected by Western blotting analysis. Results The result of MTT assay showed that compared with the control group, the cell survival ratio of DMY group, VP16 group and DMY+VP16 group reduced at 24 hours and 48 hours.The result of double dye flow cytometry showed that the apoptosis rate of the experimental groups were higher than that of the control group, and the apoptosis rate of DMY+VP16 group was significantly higher than those treated with DMY or VP16 alone.The result of the colony formation capability assay showed that,for JAR cells,the number of clones in experimental groups were less than that in the control group, and the number of clones in DMY+ VP16 group was the least. For normal liver HL7702 cells, there was no significant difference between DMY group and the control group, and there was no significant difference between VP16 group and DMY+VP16.The result of Western blotting showed that the protein expression levels of Bcl-2 and c-IAP2 in experimental group decreased compared with those in the control group, and the decrease of he protein expression levels of Bcl-2 and c-IAP2 in DMY+VP16 group were the most significant. Conclusion Dihydromyricetin combined with etoposide enhanced the inhibitory effect on choriocarcinoma cell JAR, which can reduce the dosage of the chemotherapy drug VP16.The inhibitory effect of tumor cells may be related to the down-regulation of protein expression of Bcl-2, C-IAP2. References | Related Articles | Metrics

Technology and Methodology

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Optimization of mouse in utero electroporation technique YANG Xiao-wei LIU An-jun REN An-jing WANG Zhi-BinZHANG Wei-ping Lü Jian-xin XIE Zhi-fang 2018, 49 (3):  391-394.  doi: 10.16098/j.issn.0529-1356.2018.03.020 Abstract ( )   PDF (514KB) ( )   Objective To optimize the in utero electroporation protocol for transfering genes into the embryonic murine brain. Methods Critical steps of in utero electroporation were optimized, including choosing isoflurane anesthesia, improving the preparation of micropipettes, reducing the exposure time of embryos during the surgery as well as optimizing electroporation parameters. Forty-two mouse embryos from 6 mothers were subjected to in utero electroporation at gestational day 16.5 using the optimized protocol. Results The survival rates of mothers and embryos were 100%. The tranfered gene was expressed in 85%±3% of electroporated embryonic brains. Conclusion Using the optimized in utero electroporation protocol, a satisfying survival rate of embryos and transfection efciency can be achieved. References | Related Articles | Metrics

Neurobiology

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Mechanism of metabolic disorders aggravated ischemic brain injury in tree shrews of type 2-diabetes mellitus ZHAO Ling LI Shu-qing 2018, 49 (3):  273-280.  doi: 10.16098/j.issn.0529-1356.2018.03.001 Abstract ( )   Objective To explore the mechanism of metabolic abnormality or without exacerbating ischemic brain injury by establishing tree shrew models with type 2 Diabetes mellitus (T2DM), combined with the cerebral ischemia or not. Methods Thirty-six tree shrews were randomly devided into 4 groups: control, ischemia, T2DM and T2DM combined with cerebral ischemia groups. The experimental diabetes model was established by the use of high lipid breeding combined with streptozocin (STZ) injection in tree shrews. The local cerebral thrombosis was induced by photochemical reaction in tree shrews. The metabolic status of the body was measured by serum biochemical markers. 2,3,5-triphenyl tetrazolium chloride(TTC) staining, HE staining and trunsmission electron microscopy were used to observe the changes of the body’s metabolic status at hour 24 after the most prominent clinical manifestations. Results The body weight of tree shrews in the T2DM and T2DM combined with cerebral ischemia groups was decreased (120.29±13.82)g, but there was no statistical difference(P>0.05). Fasting blood glucose (FBG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and triglyceride (TG) were significantly higher, and their values were (26.75 ± 10.60）mmol/L,（7.40 ± 3.26) mmol/L,（2.93±0.70）mmol/L,（1.93±0.63）mmol/L, respectively(P<0.05) in T2DM group, and (32.29±6.08) mmol/L, (7.80 ± 3.41) mmol/L, (3.06 ± 0.95) mmol/L and (1.73 ± 0.29) mmol/L, respectively(P<0.01) in T2DM combined with cerebral ischemia group. Serum C-reactive protein (CRP) level was obviously increased (1.43± 0.53) mg/L(P<0.01). The changes of the above indicators were more obvious when T2DM was combined with cerebral ischemia and the neurons were damaged and the area of cerebral infarction was significantly increased (19.56 ±1.25) % (P<0.01). Conclusion The metabolic abnormality of T2DM tree shrews can aggravate ischemic brain injury, and its mechanism may be related to the inflammatory response caused by the synergistic effect of CRP. References | Related Articles | Metrics

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Distribution and expression changes of the vesicular glutamate transporter 1 and vesicular glutamate transporter 2 immunopositive fibers and terminals in the postnatal development of the spinal cord of rat LI Jing CHEN Tao LI Jin-lian 2018, 49 (3):  288-293.  doi: 10.16098/j.issn.0529-1356.2018.03.003 Abstract ( )   PDF (765KB) ( )   Objective To explore different distribution and expression changes of vesicular glutamate transporter 1(VGLUT1) and VGLUT2 immunopositive (-ip) fibers and terminals in the spinal cord of postnatal 0 day (P0)-22 days (P22) rats. Methods Immunohistochemical staining was performed for the VGLUT1 and VGLUT2 in the cervical and lumber enlargements of P0-P22 rats. Results VGLUT1-and VGLUT2-ip fibers and terminals were observed in the cervical and lumber enlargements of the spinal cord in the postnatal P0-P22 rats but not soma. These fibers and termianls showed a typical complementary distribution pattern, especially in the posterior horn of the spinal cord. On P0, VGLUT1-ip fibers and terminals were mainly observed in the laminae Ⅲ-Ⅴ of the cervical and lumber enlargements, with a weak expression in the intermediate gray and anterior horn. From P3, the expression of VGLUT1 was increased in the laminae Ⅲ-Ⅴ and extended to the lateral, and the VGLUT1-ip profiles were detected in the basal part of posterior horn (lamina Ⅵ) and the lateral part of the anterior horn (lamina Ⅳ). An obvious trend of the belt distribution from the dorsomedial to ventrolateral side was observed. It was more obvious at P7, and expanded gradually to the medial and lateral side of spinal grey with development. On P22, VGLUT1-ip profiles were observed on almost whole spinal gray, except for a week expression in the lamina Ⅱ. On the contrary, VGLUT2-ip fibers and terminals were observed to distribute densely in laminae I-Ⅱ and the margins of the anterior horn from P0. In the other areas of the spinal cord, medium density of VGLUT2-ip fibers and terminals were distributed much more evenly. With the development, the distribution pattern of VGLUT2-ip fibers and terminals were not changed greatly, although its density was gradually increased. On the other hand, in the posterior cord of the spinal white matter, the developmental process of the VGLUT1 positive corticospinal posterior tract gradually went down from the cervical (P3) to the lumbar cord (P7). Conclusion The distribution of VGLUT1-and VGLUT2-ip fibers and terminals is largely different in the postnatal developing cervical and lumber spinal cord of rats, showing an obvious complementary pattern. This may be important for us to understand the difference of their functional involvement during the postnatal development of the spinal cord. References | Related Articles | Metrics

Anatomy

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Differential analysis of brain structure volume associated with Parkinson’s disease based on nuclear magnetic resonance T1 imaging LIU Gui-xue SHI Yong-hong WANG Shi-qing LI Wen-sheng 2018, 49 (3):  347-354.  doi: 10.16098/j.issn.0529-1356.2018.03.013 Abstract ( )   PDF (527KB) ( )   Objective To investigate the significance of MRI volume measurement in the morphological changes caused by Parkinson’s disease (PD) and its application in early diagnosis. The volume changes of caudate nucleus, putamen, globus pallidus, midbrain, dorsal thalamus, hippocampus and amygdala in patients with Parkinson’s disease were analyzed based on 3.0T MRI. Methods 3.0T MRI scans were performed in 40 patients with early and intermediate Parkinson’s disease (PD group) and 32 age-matched normal control subjects (NC group), respectively. The total brain, bilateral caudate nucleus, bilateral putamen, bilateral globus pallidus, midbrain, bilateral dorsal thalamus, bilateral hippocampus and bilateral amygdala, were manually delineated by the software of ITK-snap, and their volumes were calculated and normalized respectively. SPSS was used to compare and analyze the differences between the PD group and the NC group. Results There was no difference in the volume of the whole brain between the PD and the NC (P>0.05). There were significant differences in the standardized volume of the bilateral putamen, bilateral globus pallidus, bilateral dorsal thalamus, and midbrain (P<0.05), but there were no differences in the standardized volume of bilateral caudate nucleus, bilateral hippocampus and bilateral amygdale (P>0.05). Conclusion MRI volume measurement of the change of caudate nucleus, putamen, globus pallidus, midbrain, dorsal thalamus, hippocampus and amygdala in patients with PD can be helpful for the diagnosis of Parkinson’s disease. References | Related Articles | Metrics

Neurobiology

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Effects of intravenous anesthetics on lipopolysaccharide-induced proliferation of glial cells and its mechanisms DONG Chuang WANG Hao 2018, 49 (3):  299-302.  doi: 10.16098/j.issn.0529-1356.2018.03.005 Abstract ( )   PDF (359KB) ( )   Objective To detect the effect and mechanism of intravenous anesthetics on lipopolysaccharide (LPS) treated glial cells. Methods The proliferative ratio and the apoptotic ratio of glial cells after treated with LPS were detected by MTT assay, colony formation assay, and Annexin V-FITC/PI double staining, respectively. The protein related to the mechanisms was analyzed by Western blotting. Results Intravenous anesthetics reversed the LPS-induced proliferation of glial cells. The levels of p-Akt and p-mTOR were downregulated, while the levels of Caspase-3 and Caspase-9 were upregulated in intravenous anesthetics treated cells than untreated ones. Conclusion Intravenous anesthetics may reverse LPS-induced proliferation of glial cells by suppressing mTOR signaling pathway. References | Related Articles | Metrics

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Expression of ephrin A3 during valproate induced neural stem cells differentiation LI Wen HE Hui PENG Min QIN Jian-bing SHI Jin-hong JIN Guo-hua 2018, 49 (3):  294-298.  doi: 10.16098/j.issn.0529-1356.2018.03.004 Abstract ( )   PDF (470KB) ( )   Objective To investigate the expression of ephrin A3 (Efna3) in the process of neural stem cells differentiation induced by valproate (VPA) and the expression pattern of Efna3 in the nervous system. Methods The hippocampal neural stem cells of SD rats were isolated and cultured. At 24 hours and 48 hours after VPA inducement, the dynamic expression of Efna3 was detected by Real-time PCR, Western blotting and immunofluoresence assay. Real-time PCR was used to detect the expression of Efna3 in various tissues, as well as neural stem cells, astrocytes and neurons of adult SD rats. Results The expression of Efna3 increased significantly in the VPA group compared to the control group. Efna3 showed dominant expression in the cerebrum and hippocampus, and its expression was higher in neurons and lower in astrocytes. Conclusion The promotion of VPA to the differentiation of neural stem cells may be related to the upregulation of Efna3. References | Related Articles | Metrics

Cancer Biology

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Effects of annexin A5 knockdown on cell cycle of human gastric cancer MGC-803 and MKN-45 cells  WANG Xiao-jie ZHAO Yi-na LI Xin 2018, 49 (3):  342-346.  doi: 10.16098/j.issn.0529-1356.2018.03.012 Abstract ( )   PDF (400KB) ( )   Objective To investigate the effects of annexin A5 (ANXA5) konckdown on cell cycle of human gastric cancer cell lines MGC-803, MKN-45 cells. Methods Cells were diveded into interference group, negative control group and blank control group. The siRNA targeted to ANXA5 was transfected into MGC-803, MKN-45 cells by lipofectamine 2000 respectively. At the same time, negative siRNA was also transfected into negative control group and in blank control group nothing was added in. Real-time PCR and Western blotting was used to identify whether the siRNA targeted to ANXA5 can highly suppress annexin A5 expression 48 hours after transfection. When the effect of the siRNA was assured, Real-time PCR and Western blotting were applied to observe the expression of p21cip1 mRNA and P21cip1 protein, Western blotting was used to detect the expression of cyclinD1, flow cytometry(FCM) was applied to observe the cell cycle progression. Results The expression of p21cip1 mRNA, P21cip1 protein and cyclinD1 decreased significantly when compared with the negative control group and the blank control group(P<0.05). FCM analysis showed that the percentage of cells in G1 phase increased in interference group(P<0.05). Conclusion ANXA5 knockdown can down-regulate the expression of p21cip1 mRNA, P21cip1 protein and cyclinD1 in human gastric cancer MGC-803 and MKN-45 cells, it is speculated that ANXA5 may affect the development of gastric cancer by inducing cell cycle arrest by acting on cell cycle related proteins. References | Related Articles | Metrics

Anthropology

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Gene frequency on genetic characteristics of direction of eyeslits in Liaoning Xibo nationality GUO Li ZHENG Rui ZHANG Ting-shen YIN Jiao-yang 2018, 49 (3):  387-390.  doi: 10.16098/j.issn.0529-1356.2018.03.019 Abstract ( )   PDF (180KB) ( )   Objective To obtain the data of the gene frequency on genetic characteristics of direction of eyeslits in Liaoning Xibe people. Methods We used the method from “Measurement Method of Human Body” and “Anthropometric Handbook” written by Rukang Wu and Xiangqing Shao. We investigated the genetic characteristics of direction of eyeslits on 216 students (15-18 years old) who lived in Hutai Middle School, New District of Xinchengzi, Shenyang, Liaoning. We got the data of the gene frequency between the two sexes. Results According to the genetic characteristics of direction of eyeslits, the horizontal position was dominant compared with the oblique position. The dominant gene frequency of direction of eyeslits was 0.3368 in Liaoning Xibo people. The dominant gene frequency was 0.4600 in males, and 0.2528 in females. The recessive gene frequency of direction of eyeslits was 0.6632 in Liaoning Xibo people. The recessive gene frequency of dominant was 0.5400 in males, and 0.7472 in females. Conclusion There are significant differences in gene frequency distribution（P<0.01）between the male and female in horizontal and oblique position in Liaoning Xibo people. Additionally, in direction of eyeslits the frequency of dominant gene is higher than other nationality with Liaoning Xibo people in China. References | Related Articles | Metrics

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Correlation between genetic polymorphism and hypertensive disorder complicating pregnancy in Qinghai WANG Xiang-lin ZHAO De-xiong ZHUANG Wen-ting LI Hong-rong LI Jian-hua DAI Dong-fang 2018, 49 (3):  379-386.  doi: 10.16098/j.issn.0529-1356.2018.03.018 Abstract ( )   PDF (312KB) ( )   Objective To explore the correlation between the polymorphisms in endothelial nitric oxide synthase(eNOS), tumor necrosis super family member 13B(TNFSF13B), hypoxia inducible factor-1α(HIF-1α), vascular endothelial growth factor A(VEGFA) and vascular endothelial growth factor receptor 1(VEGFR1) and susceptibility to hypertensive disorder complicating pregnancy (HDCP) of pregnant women in Qinghai. Methods With the performance of a case-control study, a total of 120 Han patients with HDCP were enrolled as the study group, and 100 healthy pregnant women were included as the control group during the same period. All subjects were from Qinghai province. Sequenom MassARRAY assay was utilized to detect the genotypes of ten clinically relevant polymorphisms of HIF-1 α (RS 11549465, RS 115494657 and RS 2057482), TNFSF13B (RS 16972194), eNOS (RS 2070744), VEGFA (RS 3025029 and RS 2010963) and VEGFR1 (RS 7335588, RS 722503 and RS 12584067). Besides, the differences of single nucleotide polymorphism (SNP) sites were compared between groups subsequently. Results eNOS gene, VEGFA gene,and VEGFR1 gene were found to be significantly different between the study group and the normal group (P<0.001 or P<0.05).The proportions of genotype CC in RS 2070744 and RS 3025039 were higher in the study group than those in the control group (all P<0.001). Among them, the odd ratio (OR) of C allele in the two loci were 2.13 (1.45-3.12) and 4.95 (2.97-8.26) in the HDCP population (all P<0.001), respectively.The proportions of genotype GG in RS 2010963 and RS 7335588 were lower in the study group than those in control group (all P<0.05). Meanwhile, the OR of G alleles in the two loci were 0.50 (0.34-0.74) and 0.46 (0.30-0.72) in the HDCP population (all P<0.001), respectively. Furthermore, the proportion of RS 722503 genotype TC in the study group was 56.67%, higher than that of the control group of 29%, and the OR of C allele was 2.46 (1.58-3.84) in the population HDCP (P<0.001). Conclusion The genetic polymorphisms of VEGFA, VEGFR1 and eNOS are associated with HDCP in Qinghai province, which may be the potential genetic risk factors inducing high incidence of the disease in Qinghai. References | Related Articles | Metrics

Review

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Application of clearing methods on tissues in the three-dimensional imaging research LI Ying-ze SHAO Zhi-hua LI Si-guang 2018, 49 (3):  400-405.  doi: 10.16098/j.issn.0529-1356.2018.03.022 Abstract ( )   PDF (447KB) ( )   With the development of the microscopy and the big data acquisition processing system, the research direction gradually becomes the construction of biological structures by 3D imaging. However, because of opaque which leads to scattering of imaging light through the tissues, the depth of the imaging becomes a challenge. A technique named “clearing” can increase the organization transparency so that they can enhance the depth of the imaging and have a three-dimensional reconstruction. Being hydrophilic, the clearing reagents were divided into solvent-based clearing and aqueous-based clearing. The solvent-based clearing reduced water in the tissue and aqueous-based clearing used the hydration shell made by water to keep the fluorescent protein stability. In this article we introduced different clearing method and compared them from clearing time and retention in the fluorescence. References | Related Articles | Metrics

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Research progress of neuron specific enolase in development，injury and repair of the central nervous system ZHOU Jing-ying ZHANG Song-ou MENG Xiao-fang LIU Xue-hong 2018, 49 (3):  395-399.  doi: 10.16098/j.issn.0529-1356.2018.03.021 Abstract ( )   PDF (301KB) ( )   As a specific marker of neuron development and reconstruction, neuron specific enolase (NSE) expresses the spatiotemporal distribution in the development and repair of central nervous system. The change of NSE after central nervous system injury can reflect the pathological changes, and can adjust the level of NSE to achieve the effect of post injury repair. It is important for the repair and damage of the nervous system. In this review, we review the molecular structure, expression regulation, biological function of NSE and its relationship with the development, injury and repair of the central nervous system. References | Related Articles | Metrics

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Research progress of 0xidative stress in tissue regeneration GUO Jian-lin1 YAN Pei-shuo XU Cun-shuan 2018, 49 (3):  412-418.  doi: 10.16098/j.issn.0529-1356.2018.03.024 Abstract ( )   PDF (296KB) ( )   Reactive oxygen species play a critical role in cell proliferation, differentiation and apoptosis, oxidative stress, as a stress response, resultsed from the imbalance between the production and removal of reactive oxygen species within body. Low concentration of reactive oxygen species is favorable for cell growth and differentiation, and could be regarded as an essential signal to induce cell proliferation. It has been shown that the damaged animal organs, such as liver, muscle, heart, nerves, limb and tail, have ability to repair and regenerate. Tissue regeneration is closely associated with human disease and treatment, and oxidative stress is involved in these processes. This paper summarizes the role of oxidative stress and its mechanism in the process of tissue regeneration, which would provide the basis for the mechanism of regeneration and the treatment of human diseases especially regenerative medicine in the future. References | Related Articles | Metrics

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Advances in the effects and mechanisms of hydrogen on ischemia-reperfusion injury in diseases and organ transplantation HUANG Jun-long LIU Wen-wu SUN Xue-jun 2018, 49 (3):  406-411.  doi: 10.16098/j.issn.0529-1356.2018.03.023 Abstract ( )   PDF (290KB) ( )   Ischemiareperfusion (I/R) injury is a crucial pathophysiologic process in the development of various diseases or during organ transplantation. Hydrogen has been widely researched as a potential antioxidant in ischemia-reperfusion models of multiple diseases and organ transplantation. Herein, the authors review the current literature regarding the effects of hydrogen on ischemia-reperfusion injury and organ transplantation. Existing data on the effects and mechanisms of hydrogen on ischemia-reperfusion injury and organ transplantation are specifically reviewed and coupled with further suggestions for future work. In general, a substantial body of experimental evidence suggests that hydrogen significantly has a therapeutic effect on ischemia-reperfusion injury in models of multiple diseases and organ transplantation by inhibiting inflammatory response, oxidative stress and apoptosis through several underlying mechanisms. On this basis, further animal experiments and preliminary human clinical trial regarding the effects of hydrogen on ischemiareperfusion injury are need to be conducted for the early use of hydrogen as a drug in the clinic. References | Related Articles | Metrics