Acta Anatomica Sinica

Expression of microRNA-613 in human glioma and its effects on growth and metastasis

LI Chuan-fen JING Wen XIANG Dong-sheng LI Tao JIAO Shu-xin YANG Cui-ling MA Zhi-hong WANG Min

2018, 49 (5): 579-583. doi: 10.16098/j.issn.0529-1356.2018.05.003

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Objective To investigate the expression of microRNA-613(miR-613) in human glioma and its effect on the proliferation and metastasis. Methods Thirty human glioma patients tissues and 30 Non-glioma patients tissueswere detected by Real-time PCR, and to analysie their relationship with clinicopathological characteristics of gliomas. MiR-613mimics and unrelated sequence control(miR-613 con) were transfected into U87 cells using Lipofectamine TM2000. The expression of miR-613 in the two groups of cells was detected by Real-time PCR, the proliferation of cells was detected by CCK8 assay, and the invasion and migration of cells were detected by Transwell assay. Results The expression of miR-613 in Glioma tissues was lower than that in corresponding paracancerous tissues (P<0.05). The expression level of miR-613 in glioma tissue was negatively correlated with pathological grade and positively correlated with survival time (P<0.05). Compared with miR-613 con cells, miR-613 mimics cells proliferation, invasion and migration were significantly decreased (P<0.05). Conclusion MiR-613 can inhibit the proliferation, invasion and migration of glioma cells, and it might be a potential therapeutic target for the treatment of glioma.

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Enhancing effect of DLL3 gene silencing on the sensibility in human leukemia K562/ADM cells to adriamycin

WANG Shi-guang SI Xu-yan WANG Li-jun TONG Lei LI Yang-jie

2018, 49 (5): 624-629. doi: 10.16098/j.issn.0529-1356.2018.05.010

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Objective To investigate whether Delta-like ligand 3 (DLL) gene silencing could influence the sensibility of human leukemia K562/ADM cells to adriamycin (ADM) in vitro . Methods K562/ADM cells were stably transfected with specific shRNA interference plasmid vector targeting for DLL3. The mRNA expression level of DLL3 were measured by RT-PCR. CCK-8 assay was employed to detect the cytotoxic effect of ADM in K562 and K562/ADM cells. The apoptosis and intracellular ADM concentration were analyzed by flow cytometry. Western blotting was performed to determine the protein expression levels of DLL3, glutathione S transferases-π, (GST-π) and P-glycoprotein (P-gp). Results K562/ADM cells had a significantly higher mRNA and protein expression level of DLL3 than K562 cells (P<0.05). The IC₅₀ value of K562 and K562/ADM cells to ADM were 1.08 and 34.93 mg/L respectively. After DLL3 gene silencing, the resistant factor of K562/ADM cells declined to 13.12 with a reversal fold of 2.47. DLL3 gene silencing had no effect on apoptosis of K562/ADM cells, but a significant effect on downregulating protein levels of P-gp and GST-π (P<0.05) and increasing intracellular ADM concentration (P<0.05), and resulting in an enhance of ADM-induced apoptosis (P<0.05). Conclusion DLL3 gene silencing may enhance the sensibility of K562/ADM cells to ADM by down-regulating P-gp and GST-π, and increasing intracellular ADM concentration.

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Effects of microRNA-145 on the migration and invasion of lung cancer cell line A549 through mitogen-activated protein kinase and phosphatidylinositol 3 kinase/protein-serine-threonine kinase pathways

2018, 49 (5): 630-635. doi: 10.16098/j.issn.0529-1356.2018.05.011

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Objective To study the effect of microRNA-145(miR-145) on the migration and invasion and on the mitogen-activated protein kinase(MAPK) and phosphatiolylinositol 3 kinase/protein serine threonine kinase(PI3K/AKT) pathways in non-small cell lung cancer A549 cells. Methods The A549 cells were divided into miR-145 mimics, negative mimics (miR-NC), antago miR-145 and antago-miR control (antago-NC) groups. Transwell migration experiment and matrix matrigel invasion experiment were used to detect the influence of miR-145 on the migration and invasion ability of human non-small cell lung cancer A549. Western blotting method analyzed the effect of miR-145 on MAPK and PI3K/AKT pathways. In addition, the inhibitors of extracellular regulated protein kinase(ERK) and AKT pathways were used to detect the changes of the cell migration and invasion in A549 cell lines. Results The amount of miR-145 mimics group through transwell chambers was significantly less than the miR-NC group (175.33±23.67). The miR-145 mimics group had a lower number of cells through matrigel matrix than miR-NC group (153.33±22.33), P<0.05. When the expression of miR-145 was inhibited, the number of cells in antago-NC group through Transwell chambers or matrigel matrix were significantly less than the antago miR-145 group (245.00±23.00) and (185.00±12.00), P<0.05. The above results proved that miR-145 inhibited cell migration and invasion ability of A549 cells. The transfection of miR-145 mimics respectively inhibited the phosphorylation of ERK1/2, AKT (ser-473) and AKT (thr-308) about 90%, 78% and 73%, while the transfection of antago miR-145 promoted the phosphorylation of ERK1/2, AKT (ser-473) and AKT (thr-308) and rose up to 115%, 125%and 129%. The migration and invasion ability of A549 cells decreased when the activation of MAPK pathway and PI3K/AKT pathways were inhibited. Conclusion miR-145 regulates lung cancer A549 cell migration and invasion through MAPK and PI3K/AKT pathways, which may provide reliable research results for the treatment of lung cancer.

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Three-dimensional visualization of the mouse renal distal convoluted tubule

DENG Si-qi GU Ling MIAO Jun-ke LIAN Jie ZHAI Xiao-yue

2018, 49 (5): 636-640. doi: 10.16098/j.issn.0529-1356.2018.05.012

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Objective The renal distal convoluted tubule (DCT) is the last segment of the nephron. The study on the demarcation, as well as the spatial course and relationship between the DCT and its adjacent tubules, may lay an important structural basis for understanding the mechanism on the morphogenesis of the connection between the collecting duct and the nephrons, and contribute to the understanding on the regulation mechanism of water and salt metabolism in the DCT. The aim of this study is to visualize the course of the mouse distal convoluted tubules with the aid of the three-dimensional reconstruction technology based on the consecutive renal tissue sections. Methods Three C57/BL/6 J mice were fixed via perfusion and embedded in Epon 812. Tissue blocks were cut perpendicularly to the longitudinal axis of the kidney. A total of 720 consecutive sections with a thickness of 2.5 μm were obtained from the renal capsule to the outer stripe of the renal outer medulla. After image recordings and alignment, distal convoluted tubules from 90 nephrons were traced with the custommade computer program. Based on the recorded data, the courses and the spatial arrangement of the DCT were visualized, and the length was also measured.
Results The DCT commenced downstream from the macula densa at an average length of 40-180 μm. The shape of the cell of DCT changed abruptly from simple cubical to high columnar, and the cell nucleus was located near the apical membrane of the cell. The DCT surrounded their parent glomerulus which were located at cortical labyrinth. Virtually, the DCT did not intermingle and each tubule occupied its own separate territory in the cortex. The end of the DCT gradually transited to the connecting tubules whose cells turned short and the nuclei were not limited to the apical membrane. Usually, the DCT of superficial nephron returned and made coils on the way to touch the capsule just for one time. The length of distal convoluted tubule was 500-900 μm. Conclusion Because of the short and convoluted length, the DCT occupied a relatively small and independent space, which may facilitate precise regulation by hormone along the segment.

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Analysis of the risk factors of vertigo in the patient with the change of cervical curvature

WANG Ying YIN Chuang XU Jian-ke WEI Na TAN Jun

2018, 49 (5): 641-645. doi: 10.16098/j.issn.0529-1356.2018.05.013

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Objective To analyze the risk factors of vertigo in young and middle-aged patients with cervical curvature changes. Methods From June 2016 to March 2017, 78 young patients (32 males and 46 females) with vertigo were collected in the department of neurology of the third affiliated hospital of Xinxiang. The patients were divided into normal cervical curvature group (n=22) and abnormal cervical curvature group(n=56).The medical history and laboratory examination were compared between the two groups and were statistically analyzed. Results There were significant differences in fasting blood glucose (FBG), glycated hemoglobin (HbA1c), homocysteine (Hcy), smoking, diabetes, homocy steinemia and occupation between the two groups (P<0.05). There was no significant difference in gender and age between the two groups (P>0.05). Conclusion FBG, HbA1c, Hcy, smoking, diabetes and occupation are the common risk factors in young and middle-aged patients of vertigo with cervical curvature changes.

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Microanatomy of suboccipital retrosigmoid keyhole approach via petrosal fissure and cerebello-pontine fissure

CHANG Shu-feng YANG Bo ZHENG Lu FU Zhan-sheng YANG Wan-jing HUANG Xiao-feng WANG Jun-shan LIU Chang

2018, 49 (5): 646-650.

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[Abstract] Objective: By microanatomy?research on structures in various sections of suboccipital retrosigmoid?Keyhole via approach of petrosal fissure and cerebello-pontine fissure， so as to provide anatomy data for clinical application.Methods: 15 wet head specimens of 30 normal adults were dissected: the approach operation was simulated， the cadaveric heads were fixed onto head shelf in lateral?position， retromastoid?transverse (vertical) incision was adopted， then a bone window of 2.5cm-3cm was opened， to incise endocranium and measure change of operation field exposure scope， exposed situations of trigeminal?nerves， facial?and auditory nerves and glossopharyngeal nerves at place where such nerves enter brainstem before and after separation of petrosal fissure and cerebello-pontine fissure； petrosal fissure and ramus?inferior and ramus?superior of cerebello-pontine fissure were dissected under microscope； research objects concerninig petrosal fissure， ramus?inferior and ramus?superior of cerebello-pontine fissure， petrosal?vein， cerebellar?arteries， trigeminal?nerves， facial and auditory?nerves and glossopharyngeal nerves at place where such objects enter brainstem were measured， and photos were taken.Results: The exposed anatomical structures by this Surgical approach: upwards to tentorial edge， downwards to foramen magnum jugular tuberculum， inwards to lateral side of pons?and midbrain. Cerebellopontine?angle?area including petrosal?vein， upper， middle and lower neurovascular?territories of cerebellum were exposed. Distance between before and after separation of petrosal fissure and cerebello-pontine fissure is of statistical difference.Conclusion: This Surgical approach is the supplement and enlargement of typical retrosigmoid?approach，with small incision and less brain injury， it can fully utilize the natural space of epencephalon， so as to increase operation space on the premise of not pulling or less pulling epencephalon； this Surgical approach is operable in anatomy in microvascular decompression treatment of cranial nerve disease，this Surgical approach can make the structures in relevant area of posterior cranial fossa more exposed， to provide anatomy space for excision of space-occupying lesions in cerebellopontine angle area.

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Transcription and regulation of rDNA in cell development

LI Jia WANG Qi GONG Tian-tian ZHANG Yue SHAN Zhi-yan LEI Lei

2018, 49 (5): 695-698. doi: 10.16098/j.issn.0529-1356.2018.05.023

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The growth and development of cells are regulated by ribosome DNA(rDNA) and protein gene. Eighty-five percent of the total RNA is the transcription product of rDNA. Changes in rDNA transcription may lead to ribosomal biosynthesis stress, which affects cells growth, proliferation and differentiation, and further leads to the occurrence of diseases. Revealing the mechanism of rDNA transcriptional regulation is helpful to further elucidate the mechanism of cell development and find a new target for the treatment of diseases. In this paper, we describe the structure of rDNA and its transcriptional regulation,and discuss the roles of rDNA on the stem cells and occurring, treatment of disease, which provides a theoretical basis for rDNA in clinical research.

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Histogenesis of neuroimmune in the mouse retina

LI Rui-ping XU Gao-lei SUI Yi-zheng WANG Lai FAN Wen-juan DENG Jin-bo

2018, 49 (5): 561-570. doi: 10.16098/j.issn.0529-1356.2018.05.001

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Objective To investigate the histogenesis of neuroimmune system, such as microglia and blood retinal barrier (BRB), in the mouse retina. Methods Immunofluorescent staining, DiI diotistic assays, gelatin-ink perfusion and transmission electron microscopy (TEM) were used to visualize the structures of retinal microglia and BRB of Kunming mice at various ages (n=5-10). Results At as early as embryonic day 10(E10), the microglia distributed over retina evenly. With development, the microglia changed from an amoeba-like shape to a star-like shape with many processes. The number of microglia increased after the birth and reached their plateau at postnatal day 5 (P5), then felled down after P30. After birth, the retinal vasculature developed from the optic disk, and grew out to entire retina at P10. As age increasing, the volume density of vasculature was declined. The blood retinal barriers appeared as early as at P0. At P30, they became mature, consisting of endothelial cell, basal membrane, pericyte and the terminal feet of astrocytes. Conclusion As retinal development, microglia become more and more mature, and their number changed with parabolic model. At P30, the BRB have been developed maturely in the structures with endothelial cell, basal membrane, pericyte and the terminal feet of astrocytes. The microglia and BRB are involved in anti-infection and resistance to various pathogens.

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Brain barrier structure of APPSWE Tg2576 mice

CUI Zhan-jun LIU Fang ZHAO Kai-bing LI Bing-mei LIU Zhong-hua

2018, 49 (5): 571-578. doi: 10.16098/j.issn.0529-1356.2018.05.002

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Objective To investigate the basic structure of the blood brain barrier (BBB) and blood cerebrospinal fluid barrier (BCSFB) in mice and their changes in structure, function, and ultrastructure during the development and progression of AD. Methods The APPSWE Tg2576 mice were used and divided into APPSWE transgenic positive mice (model group) and littermates wild type mice (control group), twety mice in each group. After 16 months of feeding, whole body perfusion was performed and the craniotomy was performed to obtain the lateral ventricle wall and its choroid plexus. Immunofluorescence and transmission electron microscopy were used to observe the ultrastructure of BBB and BCSFB, so as to observe the changes of brain barrier of AD model. Results The vascular density was significantly lower in the AD model group than in the control group; the normal structure of the brain barrier in AD mice was impaired, mainly due to the connection between the brain vascular endothelial cells (or choroid plexus endothelial cells) and their organelles being damaged. The ultrastructure of the choroid plexus also showed significant changes. The main manifestations were the widening of the intercellular space, and some of the connecting structures between the cells, such as adhesion and connection, and some vesicle-like structures in the cytoplasm. Conclusion Compared with normal mice, the brain barrier of AD rats is damaged, which may lead to corresponding changes in the brain barrier transport mechanism and affect the clearance of Aβ in the brain, and the steady-state mechanisms existing in the brain barrier, such as secretions and receptors thereof. Mediated signaling may also change, and these factors may be involved in the formation and progression of AD.

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Role of mammalian target of rapamycin in brain physiology and pathology

ZHOU Xiang FU Wei-da CHEN Zhi-chi WANG Tong-tong YE Xing ZHOU Peng CUI Huai-rui SUN Chen-you

2018, 49 (5): 688-694. doi: 10.16098/j.issn.0529-1356.2018.05.022

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Target of rapamycin（TOR）and its mammalian ortholog mammalian target of rapamycin(mTOR) have been discovered in an effort to understand the mechanisms of action of the immunosuppressant drug rapamycin. mTOR is a serine/threonine kinase existed in two functionally distinct complexes, mTORC1 and mTORC2, which control many basic cellular functions such as protein synthesis, energy metabolism, cell size, lipid metabolism, autophagy, mitochondria and lysosome biogenesis. In addition, mTOR-controlled signaling pathways regulate many integrated physiological functions of the nervous system including neuronal development, synaptic plasticity, memory storage, and cognition. Thus it is not surprising that deregulation of mTOR signaling may involve in many neurological and psychiatric disorders. Preclinical and preliminary clinical studies indicate that inhibition of mTORC1 can be beneficial for some pathological conditions such as epilepsy, cognitive impairment, and brain tumors, while a direct or indirect s timulation of mTORC1 can be beneficial for other pathologies such as depression or axonal growth and regeneration.

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Effects of microRNA-25 on cerebral ischemia/reperfusion injury-induced cell apoptosis

ZHANG Jun-feng ZHAO Zhao-hua HAO Jia-hui FANG Zhi-chao XU Cang-bao XU Xi

2018, 49 (5): 584-590. doi: 10.16098/j.issn.0529-1356.2018.05.004

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Objective To evaluate the role and the possible mechanisms of microRNA-25 (miR-25) in regulating oxygen-glucose deprivation (OGD)/reperfusion (OGDR)-induced cell apoptosis in cultured SH-SY5Y and IMR-32 cell. Methods The OGD model was constructed in human SH-SY5Y and IMR-32 cells to mimic I/R injury. Fragments mediated miR-25 overexpression, mi-R25 mimics, were synthesized and constructed into recombined lentivirus plasmid. Using lipofectamine, the plasmids were transfected into the cells. The cell viability and apoptosis were detected with MTT assay and TUNEL. The mRNA and protein expression of the target genes were detected using RT-PCR, Real-time PCR and Western blotting respectively. Results Compared with the normal group, miR-25 was down-regulated in OGDR model group with decreased cell viability and increased apoptosis. Meanwhile, mRNA and protein expression of Bax and Caspase-3 were increased, but with decreased expression of Bcl-2 (n=3, P<0.05). Compared with the OGDR group, cell viability was increased and the apoptosis was decreased in miR-25 transfection group, with decreased expression of Bax and Caspase-3, as well as increased expression of Bcl-2 (n=3, P<0.05). Conclusion Taken together, our result indicate that up-regulation of miR-25 inhibits cerebral I/R injury-induced apoptosis through regulating Bax/Bcl-2 and Caspase-3 signaling pathway, which will provide a promising therapeutic target for cerebral ischemia/reperfusion injury.

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Methylation of endothelin receptor B gene in four breast cancer cell lines and its effect on MCF-7 cell proliferation

LIU Li-kun ZHU Wen-bin LIU De-shui LI Xin LIN Yue-ming ZHANG Wei ZHOU Li YUE Li-ling

2018, 49 (5): 611-616. doi: 10.16098/j.issn.0529-1356.2018.05.008

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Objective To investigate the expression level and methylation status of endothelin receptor B (EDNRB) gene in four breast cancer cell lines and the effect of re-expression of methylated EDNRB on MCF-7 cell proliferation. Methods Methylation-specific PCR (MS-PCR) and bisulfite sequencing PCR (BSP) were used to detect methylation status of EDNRB in breast cancer cell lines. EDNRB gene expression level was measured by reverse transcription-PCR (RT-PCR). The colorimetric MTT assay and colony formation were used to investigate the effect of re-expression of methylated EDNRB on MCF-7 cell proliferation. Results No gene expression and promoter hypermethylation of EDNRB were detected in MCF-7 and ZR-75-1 cell lines. While the low frequency of methylation event happened in MDA-MB-231 cells which had the highest expression level of EDNRB. These results suggested that, in 4 breast cancer cell lines, the negative correlation existed between methylation status in the promoter region and expression of EDNRB gene. The expression of EDNRB gene in MCF-7 cells could be recovered by 5-azacytidine (5-Aza-CR) in a dose-dependent manner from 5 μmol/L to 10 μmol/L. Re-expression of EDNRB gene significantly inhibited cell proliferation. Conclusion Methylation of CpG islands in the promoter region of EDNRB gene may play important roles in the carcinogenesis and development of breast cancer. EDNRB gene may be a new molecular marker in the early diagnosis of breast cancer.

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Effect of adverse psychological stress on the growth and epithelial growth factor receptor, phosphorylated protein kinase B and vascular endothelial growth factor expressions of human ovarian cancer cells transplanted into nude mice

JIN Ling-yan WANG Wan-wan ZHANG Hong CHEN Qi

2018, 49 (5): 617-623. doi: 10.16098/j.issn.0529-1356.2018.05.009

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Objective To explore the effect of adverse psychological stress on the growth and epithelial growth factor receptor(EGFR),phosphorylated protein kinase B(p-Akt) and vascular endothelial growth factor(VEGF) expressions of human ovarian cancer cells transplanted into nude mice. Methods Twenty-four female BALB/c(nu/nu)nude mice, aged 4 to 6 week-old, were randomly divided into two groups (12 mice per group): the tumor bearing group in which the human ovarian cancer tissue block was transplanted into nude mice subcutaneously without stress treatment, and the tumor bearing with stress treatment group in which the human ovarian cancer tissue block was transplanted into nude mice subcutaneously with stress treatment. After mice were sacrificed, tumors were separated and weighted. The protein expressions of EGFR, p-Akt and VEGF in the two groups were detected by immunohistochemistry and Western blotting method . Results The intensities of the immunohistochemical positive staining for EGFR,p-Akt,VEGF proteins in the tumor bearing with adverse stimulation group were higher than those in tumor bearing group. The results of Western blotting demonstrated that the intensities of the protein expressions for EGFR,p-Akt,VEGF in human bearing +stress group were higher than those in tumor bearing group. Conclusion The adverse psychological stress may promote the human ovarian cancer occurrence and development through by EGFR and its downstream PBK/AKT signal pathway.

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Influence of venous drainage position on arterial blood flow in the conjoined flap

CHENG Sheng Ⅺ Shan-shan LIN Dong-hui HE Yao-zhi XUE Bin-wei TANG Mao-lin MAO Yi-hua

2018, 49 (5): 651-656. doi: 10.16098/j.issn.0529-1356.2018.05.015

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Objective To investigate the influence of venous drainage position on the direction of arterial blood flow in the conjoined flap. Methods Seventy-two Sprague-Dawley rats were randomly divided into four groups: normal group received no treatment; the control group only remained the right intercostal posterior artery and vein; experimental group 1 only remained the right intercostal posterior artery and the left intercostal posterior vein, and experimental group 2 only remained the right intercostal posterior artery and the left iliolumbar vein. The distance among the angiosomes was measured. At day 7 after surgery, the survival rate of the flap was measured and the morphology of vessels by lead oxide-gelatin angiography was observed. Materials from the choke-area 1 was drawn, HE staining was used to compare the diameter expansion of the artery. Blood from the tail vein was collected, and the contents of lactate was detected. Results The distance of angiosomeⅠtoⅡ was nearest, Ⅰ to Ⅲ was farthest, the difference was statistically significant (P<0.05). At day 7 after surgery, the survival rate of the control group was (86.5±4.3) percent, while 100 percent in experimental groups 1 and 2. Angiography showed that the number of blood vessels was relatively small in control group, larger in experimental group 1, and the largest in experimental group 2 in regions of angiosome Ⅲ and Ⅳ. Compared to the normal group, the diameter of the artery of the experimental 2 was most obviously expanded, followed by the experimental group 1, the difference was statistically significant (P<0.05). The lactate contents in the control group was the highest, the difference was statistically significant (P<0.05). Conclusion Changes in position of venous drainage can result in the changes of the effect of arterial perfusion and venous drainage, and then alter the vascular rete, improve the flap survival.

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A rare case of unilateral complete absence of rectus abdominis in adult male

JIA Zi-qi WANG Yu-chen ZHANG Miao-yan ZHAO Sen SHEN Xin-hua CAO Cheng-gang MA Chao

2018, 49 (5): 657-659. doi: 10.16098/j.issn.0529-1356.2018.05.016

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Objective To report a rare case of unilateral complete absence of rectus abdominis muscle in an adult male without any other malformations, and reveal the abdominal dissection process, abdominal wall muscles and their blood supplies. Methods The abdominal wall muscles, their blood supplies, and innervations were dissected and observed. The development and morphology of rectus abdominis on both sides were compared. Results The cadaver had no rectus abdominis muscle on the right side. The superior and inferior epigastric arteries met with each other and remained intact in the substitutional connective tissue within the rectus sheath. The intercostal and subcostal nerves (T7-T12) pierced the sheath and reach the superficial fascia. The left rectus abdominis muscle was complete with compensatory hypertrophy. No scar or signs of surgery were found on the skin. No abdominal organ was absent, and no adhesion was found in the peritoneum except the liver area. Conclusion The absence of right rectus abdominis may result from abnormal embryonic development before the 6th week. Rectus abdominis flaps are used in many surgical reconstruction surgeries. The knowledge of the related mutations is clinically significant for preoperative preparation and the use of rectus abdominis flap.

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Anatomy for osteotomy of mandibular angle hypertrophy

CHEN Ka-na SHAO Zheng-ren FAN Hua LI Li MARIYA M. Ei Akka wi YAN Ling

2018, 49 (5): 660-665. doi: 10.16098/j.issn.0529-1356.2018.05.017

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Objective To dissect soft tissue structure around the mandibular angle and the mandibular nerve canal accurately, as to provide anatomical reference for the mandibular angle hypertrophy operation. Methods Twenty formalin-fixed adult head-and-neck specimens were observed and measured for facial arteries and veins, the marginal mandibular branch of the facial nerves, and the vessels and nerves of masseter muscle. Five lines positioned according to the characteristic structures of the mandible were used for the measurement of mandibular nerve canal. The safety range of the soft tissue structure and the mandibular nerve canal，external carotid artery, maxillary artery, and transverse facial artery were calculated. Results The distance between the facial artery and the mandibular angle is 23.18-36.28 mm. Facial artery to the anterior margin of masseter muscle is -9.51-9.27 mm. Separation between the facial vein and the mandibular angle is 17.79-32.03 mm. The marginal mandibular branch of the facial nerve is close to the mandibular angle and the lower edge of the mandible, measurements showed a distance of -8.57-10.70 mm towards mandibular angle. At anterior masseter muscle, separation between mandibular branches and the lower edge of the mandible is-8.83-11.06 mm. The four major sources of masseter arteries are the masseter branch of the facial artery, external carotid artery, maxillary artery, and transverse facial artery. The masseter nerve entered the masseter muscle alongside with the masseter artery. In the five cross sections, the disctance from mandibular nerve canal to the lateral mandibular point is more than 10.50 mm, 14.72 mm, 15.60 mm,8.53 mm and 6.74 mm, respectively. Conclusion Attention should be paid, when undermining soft tissue in the mandibular angle, three main structures specifically the facial artery and the marginal mandibular branch of facial nerve located at the anterior aspect of mandibular muscle, and the mandibular marginal branch of the facial nerve located at the mandibular angle are prerogative. Layers removed from the masseter muscle are best done at the thickest region of the middle inferior aspect. A guide safety line can be drawn quickly during osteotomy on the buccal bone palate of the mandible based on the safety range of mandibular nerve canal.

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Comparison of femoral neck cross-sectional morphology between gorilla and human

DU Bao-pu ZHANG Li-zhao ZHAO Ling-xia

2018, 49 (5): 666-670. doi: 10.16098/j.issn.0529-1356.2018.05.018

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Objective To analyze the mechanical effect of different modes of locomotion on the whole femoral neck of the gorilla and human. Methods The femoral specimens of the gorilla（n=8）and human（n=18）were scanned by the computed tomography. We obtained five femoral neck slices from the base of the femoral head to the trochanteric line, and measured the crosssectional geometric properties. Results Cross-sectional geometric properties ［eg, Ix/Iy, Zx/Zy and standardized cortical area(CA)］ of the femoral neck were increasing from the proximal to distal portion, besides the standardized polar section modulus(Zp) of the human. The values of standardized CA and Zp were lower in the gorilla than the human. Conclusion The similarity and difference of cross-sectional geometric properties between the gorilla and human femoral neck, respectively, correspond to its structure character which acts as a cantilevered beam and the different load conditions during bipedal andquadrupedalwalking. Additionally, the polar section modulus of human mid-neck is significantly lower than other cross-sections, making it a fracture-prone region.

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Differences of bone mineral density and change with age among female adults of minorities in Guangxi

CHEN Xing-cai LI Yan YU Hong-rong GONG Jian-gu ZHOU Li-ningGONG Ji-chun XU Lin DENG Qiong-ying

2018, 49 (5): 683-687. doi: 10.16098/j.issn.0529-1356.2018.05.021

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Objective To explore the differences of bone mineral density as well as prevalence of osteoporosis(POP) and their changes with age in Maonan, Mulam, Yao and Hmong females adults in Guangxi. Methods Totally 860 female volunteers from Maonan, Mulam, Yao and Hmong in Guangxi province were enrolled in this study, and bone mineral density was measured via quantitative ultrasonic bone densitometer on right calcaneus of participants. Results Obvious statistic differences of bone quality index(BQI), T-score and POP were not noticed among Maonan, Mulam and Hmong women but were found between Yao and the other three ethnicities (P<0.05). With aging, value of bone mineral density decreases markedly from its peak in age 35-45 group in Yao and Maonan. And value of bone mineral density in Mulam and Hmong keeps steady prior to age 45 years while declines sharply after age 45 years. And in age 45-55 group, BQI and T-scoreof Yao females were significantly higher than that of Maonan, Mulam and Hmong females (P<0.05). Furthermore, in age 45-55 group, the POP of Yao women was pronouncedly lower than that of Maonan, Mulam and Hmong women but no differences revealed among Maonan, Mulam and Hmong women. Conclusion The differences of bone mineral density and POP were noticed remarkably in different ethnic women in Guangxi, and Yao women showed the best bone quality among four minorities.

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Dermatoglyphics ridge line direction of the palms of Macaca mulatta

WANG Feng-chan

2018, 49 (5): 671-676. doi: 10.16098/j.issn.0529-1356.2018.05.019

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Objective To determine the characteristics of dermatoglyphics ridge line direction of the palms of Taihang Macaca mulatta. Methods A total of 52 (37 females and 15 males) Taihang Macaca mulatta were examined in this study.The model of dermatoglyphics ridge direction of the palms was established, and compared with Macaca fuscata. Results At the population level, in interdigitalⅡ, the end of dermatoglyphics ridge lines were mainly in area 2, 5 and 6, and most frequently ended in area 6. In interdigital Ⅲ, dermatoglyphics ridge lines ended in area 6. In hypothenar 1, dermatoglyphics ridge lines were in 12/4/2 direction. There was no statistical difference when left and right palms(P>0.05) were compared. The two sides were symmetrical. Significant difference was detected between female and male Macaca mulatta in ridge line direction of palm angle 1(P<0.05), but not in that of palm angle 7(P＞0.05) or Palm angle 8 (P＞0.05). The dermatoglyphics ridge line direction of palms showed significant difference between Taihang Macaca mulatt a and Macaca fuscata (P<0.01). Conclusion Dermatoglyphics ridge line direction of the palms is one of the criteria characteristics to detect interspecific differences.

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Environmental and genetic effects on H plane in cephalograms of female twins from six to twelve years old

ZHAO Chan-yuan WANG Yang-yang WANG Mei-yan HUO Feng WANG Peng

2018, 49 (5): 677-682. doi: 10.16098/j.issn.0529-1356.2018.05.020

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Objective To study the relative effects of environment and genes on the soft tissue H plane during the mixed dentitions, and to provide theoretical evidence to establish the correct teeth treatment plan. Methods The lateral cephalograms of one hunded and eighty-three female twins were studied. The environmental and genetic effects were analyzed on soft tissue profile by twin method . Results The influence of environmental factors was detected by measurement items of soft tissue nasal roots (H1), nasal vertex (H2),subnasal point (H4),lower lip pits（H9）, and soft tissue chin point (H10). The influence of environmental and genetic factors was detected by measurement items of lower lip pits（H9). Conclusion The upper and lower lips are detected to be influenced by environmental factors, whereas the lower lip pits are mainly by the environmental and genetic factors.

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Establishment of monoclonal cell strain stably expressing α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor subunit GluA1

LI Jiong ZHANG Ji-feng ZHAO Bo CAI Zhen-bin ZHU Xiao-nan GUO Guo-qing

2018, 49 (5): 591-597. doi: 10.16098/j.issn.0529-1356.2018.05.005

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Objective To build a cell line stably expressing α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid(AMPA) receptor GluA1 subunit. Methods The GluA1 gene was amplified by PCR and loaded into the lentiviral expression vector PLV.0-green fluorescent protein(GFP). HEK293T cells were transfected by using the four plasmid system. Puromycin was used for screening stable cell lines. The monoclonal cells were identified by Real-time PCR, Western blotting, immunofluorescence and whole cell patch clamp. Results The amplified product identified by colony PCR was consistent with the molecular weight of GluA1 (2724 bp). Further sequencing result showed that the sequence inserted into the lentiviral vector was consistent with the GluA1 sequence; HEK293T stable cell lines screened by puromycin selection were confirmed by Real-time PCR, Western blotting and immunofluorescence. GluA1 expression was correct in the experimental group, but not expressed in the control group. Immunofluorescent staining showed that the expression of GluA1 protein was relatively uniform on each cell surface. Stable cell lines were detected in voltage clamp mode with 10 mmol/L glutamate in extracellular fluid, and no electrical signal was detected in the empty vector group, whereas 5-40 pA electrical signal was recorded in the GluA1 overexpressing group. Conclusion We successfully established a HEK293T monoclonal stable cell strain expressing GluA1 subunit.

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Effect of N-cadherin overexpression on protrusion formation and catenins expression in mouse neuroblastoma N2a cells

ZHU Shao-yi GUAN Li-hong YANG Ci-qing ZHANG Bi-chao LI Qiu-ling LIN Jun-tang

2018, 49 (5): 598-604. doi: 10.16098/j.issn.0529-1356.2018.05.006

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Objective To analyse the effects of N-cadherin overexpression on cell morphology and catenins expression by using mouse neuroblastoma N2a cells. Methods The mouse N-cadherin gene was cloned to pCAG-MCS-EGFP plasmid and transfected into N2a cells by Lipofectamine 3000. The empty vector pCAG-MCS-EGFP and the overexpression vector pCAG-N-Cad-EGFP transfected group were used as control group and experimental group, respectively. Cell scratche and cell adhesion experiments were used to detect the effects of N-cadherin overexpression on the migration and adhesion of N2a cells. Immunofluorescence was used to detect the effect of N-cadherin overexpression on the morphology of N2a cells. The expression of catenin family was detected by Western blotting. Results Compared with the control group, overexpressing N-cadherin promoted the protrusion formation of N2a cells, which led to the increase and elongation of protrusion in N2a cells. N-cadherin overexpression enhancea cell adhesion and inhibited migration. N-cadherin overexpression resulted in the expression level of α-catenin, β-catenin and p120 protein significantly increased (P<0.05), while γ-catenin expression decreased but the difference was not significant (P>0.05). In addition, the cell aggregation phenomenon was observed when N-cadherin overexpression. Conclusion N-cadherin overexpression promotes protrusion formation and adhesion ability in N2a cells, but inhibits cell migration, regulates the expression of the family of catenin and promotes cell aggregation.

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c-Jun N-terminal kinase mediated apoptosis and autophagy is involved in osteocyte death induced by tricalciumphosphate wear particles in the mouse calvaria prosthesis

CHEN Yang NING Le DU Jia-li ZHOU Chun-yan LI Han-xin ZOU Jia-jun FU Shi-jie ZHANG Yun

2018, 49 (5): 605-610. doi: 10.16098/j.issn.0529-1356.2018.05.007

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Objective To investigate whether c-Jun N-terminal kinase(JNK)-mediated apoptosis and autophagy is involved in osteocytes death induced by tricalciumphosphate(TCP) wear particles in the mouse calvaria. Methods Thirty-six 6-week ICR male mice were randomly divided into three groups: normal control groups (n=12), TCP wear particles groups (n=12) and SP600125-treated groups (n=12). A murine calvarial model of osteolysis was established by 30 mg of TCP wear particles implantation onto the surface of bilateral parietal bones following removal of the periosteum. On the second postoperative day, the SP600125-treated mice were locally injected with the specific inhibitor of JNK pathway (SP600125, 1.0 mg/kg) on the the calvaria under the periosteum each time every three days. After 2 weeks of continuous treatment, the animals were sacrificed and the calvaria were obtained. Calcein-AM labelling and HE staining were used to observe the morphology and viability of calvaria osteocytes; flow cytometry was performed to detected apoptosis of calvaria osteocyte obtained by enzyme digestion; Western blotting was used to examine protein expression of Bcl-2, Bax, p-JNK, Beclin-1 and LC-3 in calvaria osteocytes, respectively. Results Compared with the control group, the viability of calvaria osteocytes was significantly reduced, leading to the obvious increases in the percentage of empty lacunae and osteocyte apoptosis in the TCP group (P<0.05); protein expressions of p-JNK, Beclin-1 and LC-3, and the conversion of LC3-Ⅱ from LC3-Ⅰ were both increased remarkably (P<0.05). Compared with the TCP group, the percentage of apoptosis in calvaria osteocytes decreased, and expression of autophagyrelated protein was obviously inhibited in the SP600125 group (P<0.05). Conclusion JNK-mediated apoptosis and autophagy is involved in osteocytes death induced by TCP wear particle in the calvaria.

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