Acta Anatomica Sinica

Establishment and identification of radioresistant non-small cell lung cancer model

QI Shu-ya SHI Han-fei SHI Meng-jie ZHANG Meng-hui ZUO Yan-zhen SUN Da-yong

2017, 48 (2): 156-159. doi: 10.16098/j.issn.0529-1356.2017.02.006

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Objective To establish a radioresistant model of non-small-cell cancer A549, and to provide the experimental basis for further researchonradioresistance.Methods The cell number and the dose of radiation therapy were confirmed. The intermittent radiation were used to induce the radioresistant cell model. The morphology of cells were observed with an inverted phase contrast microscope. Colony formation was used to identify the radioresistance of the RA549 cell.Results RA549 cells were longer and bigger than A549 cells; surviving fraction of RA549 was significantly increased than A549. Conclusion Intermittent radiation can successfully induce the radioresistant cell model of lung cancer cells.

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Role of microRNAs in liver regeneration

ZANG Xia-yan GENG Xiao-fang ZHANG Chun-yan XU Cun-shuan

2017, 48 (2): 230-235. doi: 10.16098/j.issn.0529-1356.2017.02.020

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microRNAs (miRNAs) are a group of endogenous, small, noncoding RNA molecules which can regulate gene expression at posttranscriptional level by binding to the 3’ untranslated regions (3’UTRs) of their target mRNAs. It has been reported that miRNAs regulated a variety of biological processes, including cell proliferation, differentiation, apoptosis, metabolic process，organ development，etc. Liver as one of the organs with extraordinary regenerative abilities, is considered to be an important material for the research of regeneration mechanism. This paper summarized the research advance of the role of microRNAs in liver regeneration.

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Compatibility between adipose derived stem cells of rats and three dimensional printing gelatin scaffold

TIAN Xiao-hong ZHANG Bin FANG Yan BAI Shu-ling AO Qiang TONG Hao

2017, 48 (2): 209-216. doi: 10.16098/j.issn.0529-1356.2017.02.016

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Objective To observie the compatibility of rat adiposederived stem cells (ADSCs) and three dimensional printing (3DP) gelatin scaffolds crosslinked with glutaraldehyde, the most suitable for cell growth aperture, in order to provide experimental basis for constructing tissue-engineered tissues or organs. Methods The enzyme digestion method was adopted to separate and extract rat ADSCs. Flow cytometry and multidirectional differentiation method were used for identification. The ADSCs were cultured with 3DP gelatin scaffolds of different aperturs. Scanning electron microscopy (SEM) and transmission electron microscope (TEM) were used to observe the ultra-microstructure. The cell vitality analyzer was used to detect its survival rate. MTT method was used to detect the effects of two dimensional (2D) and three dimensional (3D) cultural methods on ADSCs cell vitality. Results The seeding ADSCs had multi-directional differentiation potential, and had the basic characteristics of stem cells. After inoculation of ADSCs to 3DP gelatin scaffolds, cells were oval or spindle observed by SEM, which was different from the traditional 2D culture form. The cells scattered within the scaffold gap, the structures such as the nucleus and organelles were clear, which demonstrated a good compatibility with scaffold. The cell survival rate of 90μm aperture scaffold was the highest, and 3D cultural method was more advantageous to maintain the vitality of ADSCs. Conclusion The compatibility of ADSCs with 3DP gelatin scaffold is good, and 90μm aperture scaffold is most suitable for the growth of ADSCs.

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Construction of Pim-1 recombinant adeno-associated virus 2 vector and its infection in rat retina

ZHANG Shou-mei HUANG Ting-ting WANG Dong LIU Fang XU Jia-jun

2017, 48 (2): 121-127. doi: 10.16098/j.issn.0529-1356.2017.02.001

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Objective To construct a recombinant adeno-associated virus 2 vector (rAAV2-Pim-1) carrying rat protooncogene Pim-1, and to detect infected cell types and the expression of Pim-1 in rat retina infected by rAAV2-Pim-1. Methods The pAOV-CAGMINI-EGFP-2A-MCS-3FLAG vector and Pim-1 PCR product were cleaved by Nhel enzyme. The recovered vector and target gene DNA were identified after agarose gel electrophores and linked to transformation, positive plasmid cloning and sequencing analysis. rAAV2-Pim-1 expression plasmid pAOV-CAGMINI-EGFP-2A-Pim-1-3FLAG, packaging plasmid pAAV-RC and helper plasmid pHelper were co-transfected into 293 cells through using the Lipofectamine 2000, then high titer of rAAV2-Pim-1 was purified. After rAAV2-Pim-1 injection into rat intravitreal body, the infected retinal cell types were detected by immunofluorescence histochemistry staining; the expression of Pim-1 in the retina was quantified by Real-time PCR and Western blotting. Results The construction of rAAV2-Pim-1 plasmid was successful and the nucleotide sequence was right; Green fluorescence in 293 cells was found after plasmid transfection into 293 cells; The virus titer was 5.7×10¹⁵vg/L. After rAAV2-Pim-1 infected rat retina in vivo, the infection percentage of retinal ganglion cells (RGCs) reached 71%, and a few of amacrine cells and hardly astrocytes were infected; the expression of Pim-1 mRNA and protein in the retina of rAAV2-Pim-1 group was about 6.61 times and 2.29 times of the rAAV2-EGFP group respectively.Conclusion rAAV2-Pim-1 virus vector is successfully constructed and Pim-1 is overexpressed in the RGCs of infected rat retina.

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Screening the differentially expressed genes after spinal cord injury by RNA-Seq and subsequent functional analysis for partial candidate genes

CHEN Ying WANG Di NIU Xue-ying WANG Xiao-dong CHEN Xue

2017, 48 (2): 128-134. doi: 10.16098/j.issn.0529-1356.2017.02.002

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Objective We apply RNA-Seq technology to characterize the temporal changes in global gene expression after spinal cord injury (SCI) in rats. Methods Spinal cord contusion injury was produced with the Infinite Horizon Device. A total of 48 rats were randomly divided into sham control, and contusion injury for 1 day, 4 days and 7days. RNA-Seq technology was carried out to screen the differentially expressed genes (DE genes) after SCI. We also performed expression pattern and pathway analysis for the DE genes, and selected the candidates to further expression variation validation. Results Compared with sham group, there were 944DE genes at the first day, 1362 DE genes at the 4th day and 1421 DE genes at the 7th day. The expression variation patterns were roughly divided into 8 kinds of forms.In addition, Real-time PCR results showed that the expression patterns of heme-oxygenases 1（Hmox1）, Plau, Serpine1 and Ncf2 were consistent with RNA-seq analysis. The result of immunohistochemistry showed that Hmox1 was highly expressed in spinal cord neurons after injury. Conclusion RNA-Seq analysis is useful to screen the DE genes after SCI, and the validated genes could partially explain the molecular mechanism of SCI.

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Mechanism of ischemic postconditioning relieved brain edema and cerebral infarction after cerebral ischemia in tree shrews

ZHANG Chuan-rao YU Zhi-cheng LI Shu-qing

2017, 48 (2): 135-141. doi: 10.16098/j.issn.0529-1356.2017.02.003

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Objective To observe the effects of ischemic postconditioning(PC) on changes of cerebral water content, cerebral blood flow, infarct area and hippocampal ultrastructural , and to explore the neuroprotective mechanisms of ischemic PC on undergoing thrombotic cerebral ischemic injury. Methods Tree shrews were randomly grouped into control, ischemia 4 hours，ischemia 24 hours，ischemic postconditioning 4 hours and ischemic postconditioning 24 hours (n=8) Eight animals were used for HE staining(n=3)and electron microscopy(n=5). The model of thrombotic cerebral ischemia was induced by photochemistry in tree shrews. Four hours after the model establishment, the common carotid artery on the ischemia side was clamped for 5 minutes, then perfused by removing the clamp for 5 minutes, and repeated the same management 3 times, so that the model of PC was established. The changes of the brain water content of local cortex were measured by Elliott dry-wet weight and the brain infarct area was determind by 2,3,5-triphenyl-tetrazolium chloride staining.In addition, the regional cerebral blood flow of local cortex was measured by laser doppler, and the ultrastructural changes in the CA1 area of hippocampus in different groups were observed under an electron microscope. Results More neuron pycnosis was observed in hippocampal CA1 area. Obvious swelling of mitochondria, partial disrupt and vanish of the mitochondria cristae and more endoplasmic reticulum cisterna appeared in the neuron of hippocampus at the 24th hour after cerebral ischemia.With the time prolonging of ischemic, the brain water content was significantly increased (86.81%±1.08%) in the ischemia group at the 24th hour compared with sham group. Cerebral infarction area was maximum (33.00%±3.03%) and regional cerebral blood flow decreased obviously［(134.27±28.75) ml/min］. The brain water content was significantly decreased (81.04%±1.04%, P<0.01) and the infarct area was significantly shrink in ischemic postconditioning group (16.79%±1.29%, P<0.01) than that in ischemia group.The regional cerebral blood flow in ischemic postconditioning group was in contrast to ischemia group significantly at the 24th hour ［(195.25±21.18) ml/min, P<0.01］. Conclusion Ischemic postconditioning attenuates the edema in ischemic brain and narrow the cerebral infarction area in tree shrews. The mechanism may be related to the improvement of local cerebral blood flow.

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Changes of mitofusin-2 expression after experimental subarachnoid hemorrhage in rats

YANG Xiao-mei ZHANG Yan SUN Juan PENG Shu-chen ZHOU Chang-man CHEN Chun-hua

2017, 48 (2): 142-149. doi: 10.16098/j.issn.0529-1356.2017.02.004

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Objective To investigate the dynamic changes of mitofusin-2(Mfn2) expression in rat cerebral arteries after experimental subarachnoid hemorrhage (SAH) and to reveal the relationship between Mfn2 and cerebral vasospasm (CVS). Methods A SAH model was induced by endovascular perforation of the intracranial portion of the internal carotid artery. One hundred and forty six male SD rats were randomly divided into six groups: sham group and SAH groups which were sacrificed at different time points (24 hours, 48 hours, 72 hours, 7days and 14days). Mortality, neurobehavioral score and brain water content were measured. Histology was conducted to observe the morphological changes. Western blotting and RT-PCR were performed to measure the Mfn2 protein and mRNA changes of the major cerebral arteries at different time points after SAH. Results Blood clot surrounded the basilar artery gradually dissipated after SAH. HE staining showed that the most severe morphological vasospasm in basilar arteries was observed at the 24th hour after SAH. No positive immunohistochemical staining of Mfn2 was shown in the media layer of basilar artery at the 7th day after SAH. Western blotting showed that Mfn2 protein was remarkably increased at the 48th hour and the 72th hour after SAH compared to sham groups (P<0.05) and significantly decreased at the 7th day after SAH (P<0.05). The protein level at the 14th day after SAH was almost the same level with the sham and SAH 24 hours groups. The mRNA level changed in the same tendency as the protein level. Conclusion This study indicate that Mfn2 plays essential roles in both acute and delayed CVS which may provide a theoretical basis for understanding of the mechanism of the CVS after SAH.

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Effect and its mechanism of Juglone on invasion and metastasis ability of human cervical cell line HeLa

ZHAO Xing-yu HOU Jian-cheng JIN Lian-hai WANG Yu-qi LIU Song ZHANG Wei

2017, 48 (2): 160-164. doi: 10.16098/j.issn.0529-1356.2017.02.007

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Objective To observe the effect of Juglone on invasion and metastasis of Hela cells and explore the possible mechanism. Methods HeLa cells were cultured and treated with 10,20,50,100μmol/L Juglone for 24 hours. The morphology changes of HeLa cells were observed with an inverted microscope. The viability of HeLa cells was detected by MTT assay. The cell scratch test was used to detect cell migration ability after treatment of Juglone. The ability of cell invasion was measured by Transwell chamber. The expression of matrix metalloprateinases (MMP)-2 and MMP-9 were detected by Western blotting. Results Compared with control group, the viability of HeLa cells decreased after treatment with different concentrations of Juglone for 24 hours, and the cell morphology was changed in a dose-dependent manner. Scratch test results showed that the level of cell movement ability decreased significantly with the increase of the concentration of Juglone. Transwell invasion assay showed that Juglone had a strong inhibitory effect on the invasiveness of HeLa cells in vitro. Western blotting results showed that Juglone inhibited the expression of MMP-2 and MMP-9 protein in HeLa cells. Conclusion Juglone can inhibit the invasion and metastasis in HeLa cells, and its possible mechanism may be related to down regulating the expression of MMP-2 and MMP-9.

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Recent progress on the mechanism of energy metabolism of hypoxia-inducible factor 1-driven skeletal muscle adaptations to hypoxia

SONG Ya-qiong ZHOU Bo-jiang

2017, 48 (2): 236-240. doi: 10.16098/j.issn.0529-1356.2017.02.021

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Hypoxia-inducible factor1 (HIF-1) is a key nuclear transcription factor that regulates the oxygen homeostasis of tissue cells and widely exists in both mammals and humans. The expression and activity of HIF-1 are tightly regulated by cellular oxygen concentration. It can regulate the biological effects of cellular energy metabolism, angiogenesis, erythropoiesis, cell survival, cell proliferation and apoptosis in the physiological and pathological hypoxic-ischemic condition, so that the cells can adapt to the hypoxic environment to survive, or to apoptosis. This paper reviewed the structure and function of HIF-1 and the recent progress on the study of the mechanism of energy metabolism of HIF-1-driven skeletal muscle adaptations to hypoxia.

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Validation of a system to evaluate chronic functional joint instability rehabilitation therapy

YU Hui-xian YANG Chun-sheng ZHANG Ran HU Zhi-wei HE Lei CHEN Ya-ping

2017, 48 (2): 165-169. doi: 10.16098/j.issn.0529-1356.2017.02.008

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Objective To investigate the assessment of patients with functional ankle instability prior to and post of regular rehabilitation therapies. Methods One hundred and twenty-four patients with unilateral chronic ankle instability (51 females and 73 males, age 34-56) were received conventional rehabilitation for two month, including muscle strength, range of joint motion, balance, proprioception, etc. The visual analogue scale（VAS）scores (instant pain and pain after 500-meter walk ) were recorded. Standing time of affected limb, the star excursion balance test, static and dynamic plantar pressure were performed. Results There were significant differences in VAS pain score of instant pain(5.32±0.27 vs 1.07±0.08) and after 500 meter walk (8.79±1.78 vs 4.51±1.78) (P<0.05), and standing time of affected limb and the star excursion balance test between the data after and before training. In contrast, no significant difference was observed in the groups of static and dynamic plantar pressure. Static: normal anterior foot（1095.30±61.28）gr/cm² vs（1670.30±151.22）gr/cm²; Dynamic: normal anterior foot（1654.98±294.27）gr/cm² vs affected limb（2822.30±312.28）gr/cm² (P > 0.05). Conclusion Rehabilitation therapy can significantly improve the pain-release, the balance, etc. However, the effect on the dynamic response is not accurate. Thus, the abnormal of walking posture, the biomechanics of the foot and ankle, and biomechanics of functional ankle instability should be considered during the rehabilitation treatment.

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3D visualization of membranous labyrinth from voxel models

YANG Xiao-kai WU Shu-zhi CHEN Xiao-su ZHENG Yan-yan

2017, 48 (2): 170-174. doi: 10.16098/j.issn.0529-1356.2017.02.009

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Objective To explore three dimensional (3D) visualization of membranous labyrinth using a voxel model. Methods 3D Slicer software was used to deal with MRI images of the temporal bone, volume clipping with the model, three-dimensional display of the membranous labyrinth by both surface rendering and volume rendering. Results With the inner voxel model we explored not only the surface of inner ear，but also the membranous labyrinth inside. The file size of inner ear voxel model file was smaller than surface model file. Conclusion Inner ear voxel model is invaluable in the study of inner ear anatomy.

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Effect of assessing the right bronchus orifice preintubation by bronchoscopy on adjusting the position of right double-lumen endobronchial tube

ZHU Guang-qiu WANG Xiao-xia CAO Dong-hang

2017, 48 (2): 175-178. doi: 10.16098/j.issn.0529-1356.2017.02.010

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Objective To investigate the clinical value of examining the anatomical position of right superior bronchus orifice preintubation by bronchoscopy on adjusting the position of right double-lumen endobronchial tube (R-DLT). Methods One hundred and sixty adult patients who were scheduled to intubate with R-DLT undergoing thoracic surgery were randomly assigned into two groups (n=80 per group): experimental group and control group. All patients were received intravenous anesthetics for induction. The length of right main bronchus and the position of right superior bronchus orifice in the transverse section of right main bronchus were measured prior to intubation by bronchoscopy in experimental group. The R-DLT was inserted into right bronchus by the conventional method and then adjusted the position by bronchoscopy in two groups. Bronchoscopy was performed through the tracheal lumen to ensure that the endobronchial portion of the DLT was in the right bronchus. The blue endobronchial cuff was just seen below the carina. In addition, lifted or inserted and twisted the tube properly according to the data measured in the experimental group. The relative position of orifices of the tube and the right upper lobe bronchus was assessed by bronchoscopy in the two groups. The R-DLTwas considered as effective preliminary location if right superior bronchus orifice was partly seen through the lateral orifice of R-DLT. The position of R-DLT was further adjusted till the three orifices of segmental bronchi were seen. Results In experimental group, the length of right main bronchus was (2.29±0.58)cm which was shorter than 1cm in 2 cases (2.5%). The central angle in the transverse section of right main bronchus between the central point of right superior bronchus orifice to 12 o’clock position was (94.5±8.3)° which was smaller than 75°or bigger than 105° in 4 cases (5.0%) in total. The number of effective preliminary location of R-DLT was 77(96.3%) in experimental group, and 62(77.5%) in control group, which showed significant difference between the two groups(P<0.05). There was 1 case (1.25%) failed intubation in each group, because of short distance within 1cm between right superior bronchus orifice and tracheal carina. Conclusion Examining the anatomical position of right superior bronchus orifice preintubation is helpful to improve effective preliminary location of R-DLT, and it is beneficial to find out anatomical variation and select appropriate airway tube.

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Comparison of distribution characteristics of extracellular matrix components in the testis of the Tibetan sheep and the Small-tail Han sheep from Plateau

Ligang YUAN ZHANG Yong Cong Li CHENG Xu-dong LU Qi-zhong

2017, 48 (2): 179-186. doi: 10.16098/j.issn.0529-1356.2017.02.011

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Objective To compare the distribution and histochemical characteristics of extracellular matrix components in the testis between 4 Tibetan sheep and 5 small-tail Han sheep from plateau at the same age. Methods Histochemistry and transmission electron method were used to characterize the microstructure and ultrastructure of the two kinds sheep testis. Image-Pro Plus(IPP) statistics methods were used to identify the integrated absorbance for laminin(LN), type Ⅳ collagen(ColⅣ) and heparan sulfate proteoglycans(HSPG) distribution investigation by immunohistochemistry. Results The light microscopy observation showed that compared with the testis of the small-tail Han sheep from plateau, the collagen and reticular fibers were richer in interstitial of the Tibetan plateau sheep. The periodic acid-schiff(PAS) and Alcian blue-periodic acid schiff(AB-PAS) staining positive reactions were more obvious in interstitial basement and blood capillary of the Tibetan plateau sheep testis. Immunostaining analysis showed that the Col Ⅳ was present strongly in the seminiferous epithelium of both the Tibetan plateau sheep and the small-tail Han sheep. The expression of LN in the Sertoli cells and peritubular myoid cells of the Tibetan plateau sheep testis was weak but moderately positive in the Sertoli cells, the Leydig cells and peritubular myoid cells of the small-tail Han sheep testis. In the both kinds of sheep testis, the expression of HSPG was especially strong in the peritubular myoid cells but weak in the Sertoli cells and Leydig cells. The integrated absorbance of Col Ⅳ and LN expression was significantly lower in testis of the Tibetan sheep than the smalltail Han sheep (P<0.01）, but the average absorbance of HSPG expression was markedly higher than it (P<0.01). Electron micrographs of portion of the seminiferous tubule in the both of the testis showed elaborately developed tunicae propria with a well defined epithelial basal lamina and a distinct layer of typeⅠcollagen fibrils. The collagen fibrils were found around the interstitial blood capillaries and the abundant Lipid droplets in the Leydig cells of the small-tail Han sheep testis from plateau. Conclusion In plateau environment, the connective tissue in the tunicae propria and interstitial tissue of the Tibetan sheep testis is more abundant, and partly affect the development of the seminiferous epithelium. The AB-PAS positive reaction in interstitial basement and the wall of blood capillary in the Tibetan plateau sheep testis are closely related to the secretory function of the Leydig cells. The integrated absorbance of LN expression is significantly higher and HSPG is lower markedly in testis of the smalltail Han sheep testis, which is very important to the development degree of seminiferous epithelium.

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Inhibition effect of sorafenib on adjuvant arthritis in rats

LIU Fei GONG Yong-fang WANG Zhen-zhen ZHANG Xiao-ming DENG Xue-fei JIAO Yi HUANG Xue-ying

2017, 48 (2): 187-192. doi: 10.16098/j.issn.0529-1356.2017.02.012

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Objective To observe the inhibition effect of sorafenib on adjuvant arthritis in rats. Methods A total of 36 male SPF SD rats were divided into 6 groups and 0.1ml of the complete Freund’s adjuvant was subcutaneously injected into the left hind paw except the normal group. The volume of rat hind paw was measured. The CD4⁺ and CD8⁺T cell subsets of the peripheral blood were analyzed by flow cytometry. The microvessel density of synovial tissues was determined by immunohistochemical chain mildew avidin peroxidase enzymatic (SP) method. Results Compared with model group, the rats of sorafenib groups alleviated the volume of rat hind paw and reduced microvessel density. Sorafenib (20, 40mg/kg) groups decreased the percentage of CD4⁺T cell and increased the percentage of CD8⁺T cell at the same time. All the values were statistically significant(P<0.05). Conclusion Sorafenib can ameliorate adjuvant arthritis in rats, which effect may be related to sorafenib causing CD4+ and CD8+T cell subset of peripheral blood deviation and reducing microvessel density of synovial tissues.

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Correlation between the expression of P34H and the activity of hyaluronidase in human spermatozoa

MA Xiao-ping GAO Xiao-qin YUE Ying-quan WANG Jun

2017, 48 (2): 193-198. doi: 10.16098/j.issn.0529-1356.2017.02.013

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Objective To investigate the correlation between the level of P34H expression and the activity of hyaluronidase in human spermatozoa. Methods Eighty eight semen samples were collected, 68 cases were in the infertile group, and 20 in the normal control group. Semen routine analysis was referred to the WHO standard method. According to the difference of semen parameters, 68 cases of infertile males were divided into the infertile group with normal semen parameters and the abnormal semen parameters. Western blotting was used to detect the level of P34H expression on spermatozoa. The P34H-positive rate on human spermatozoa was determined by indirect immunofluorescent staining using anti-P34H antibody. The HYD-positive rate and HYD-activity intensity in all samples were examined by improved fixed-substrate film method. Results The level of P34H protein expression and the percentage of the P34H-positive rate in infertile groups with normal semen parameters and abnormal semen parameters were significantly lower than that in the control group (P<0.05). The activity of HYD (HYD-positive rate, HYD-activity intensity) in the infertile groups with normal semen parameters and abnormal semen parameters were also significantly lower than that in control group (P<0.05). The relation between the P34H protein expression and HYD-positive rate, HYD-activity intensity had a significant positive correlation (r=0.449, 0.431；P<0.01); the relation between the the percentage of P34H-positive rate and HYD-positive rate, HYD-activity intensity had a significant positive correlation (r=0.727, 0.691；P<0.01). Conclusion The level of P34H protein expression and the percentage of the P34H-positive rate are decreased, while the activity of HYD (HYD-positive rate, HYD-activity intensity) is reduced in male infertility.

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Use of morphology inspection methodology in the myocardial tissue samples

SUN Yang ZHAO Hong

2017, 48 (2): 225-229. doi: 10.16098/j.issn.0529-1356.2017.02.019

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Objective To investigate the use of morphology inspection methods in myocardial diseases, and to evaluate their advantages and disadvantages. Methods Myocardial tissues were staining by phosphotungstic aci hematoxylin(PTAH)，Masson，elastic fiber tissue (ET)+Van Gienson(VG)， Congo red，Schiff periodic acid shiff(PAS)，and Gram staining methods. Immunehistochemical method, and transmission microscopy had also been applied to study the myocardial tissue. Results HE staining differentiation of myocardial tissue was not well; Masson trichromatic staining differentiation of myocardial cells and collagen fibers had clear distinction; ET + VG staining method was able to distinguish among elastic fibers,collagen fibers, fibers and amyloid; Congo red stain was mainly used in the detection of amyloidosis, the PAS reaction assisted in the diagnosis of glycogen storage disease. Immunohistochemical technology was an useful tool in disease model study and diagnosis of the cardiac disease. Electron microscopic observation of the ultrastructure was an important means of etiological diagnosis. Conclusion For myocardial tissue samples, a combination of morphology methods of the histology, subcellular and molecular can get a better observation.

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Novel protein C7orf42 promotes rat hepatocyte BRL-3A proliferation

ZHANG Chun-yan CHANG Cui-fang ZHANG Shi-fu MA Ji ZHANG Fu-chun XU Cun-shuan

2017, 48 (2): 150-155. doi: 10.16098/j.issn.0529-1356.2017.02.005

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Objective To explore the effect of C7orf42 on cell proliferation of rat hepatocyte line BRL-3A in vitro. Methods The expression of C7orf42 was knocked down by siRNA, and MTT and EdU assay were used to discover the effect of C7orf42 on cell proliferation at 48 hours after transfection. Flow cytometry was used to observe the effect of cell cycle progression. Real-tme PCR and Western blotting were used to detect the changes in the expression of cell proliferation-associated gene. Results MTT results showed that the cell viability of the interference group (C7BRL-siR3) was significantly lower than that of the negative control group (NC) at 48 hours after transfection (P<0.05). Meanwhile, the percentage of EdUp-labeling cells was also significantly decreased (P<0.05). At the same time, the flow cytometry results showed that the number of cells in division phase (S+G₂/M) of the interference group was significantly reduced in parallel (P<0.05). Further, the interference group down-regulated the expression levels of cell proliferation-related genes and proteins of JUN, MYC, CCND1 and CCNA2. Conclusion C7orf42 may promote cell proliferation via regulating the expression of JUN, MYC, CCND1 and CCNA2 in rat hepatocyte line BRL-3A.

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Relationship between body composition and bone mass abnormal in Tibet Tibetans

YANG Xiu-lin HE Ye MA Li-yang HE Jin-quan HAI Xiang-jun

2017, 48 (2): 199-203. doi: 10.16098/j.issn.0529-1356.2017.02.014

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Objective To investigate the distribution of bone strength index of adult Tibetans in Tibet and analyze the correlation between of their body composition and incidence of bone mass abnormal. Methods The bone strength of right calcaneus, T score and body composition of 560 Tibetans in Shigatse, Tibet by using an ultrasonic bone mineral density meter and bioelectric impedance technique. Binary LogisticRegression was used to test the relationship of incidence of bone mass abnormal and body composition. Results The bone mass index of the male and female participants reached the maximum at their 18-30 years old, and then declined with aging. The incidence of osteoporosis among male and female participants ≥50 years old were 7.6% and 11.7%respectively. Age（B=0.046，OR=1.047，P<0.01）and visceral fat mass（B=0.452，OR=1.572，P<0.05）were risk factors of incidence of bone mass abnormal whereas the subcutaneous fat mass(B=-0.181，OR=0.835，P<0.01)and muscle mass (B=-0.055，OR=0.947，P<0.05)were protective factors. Conclusion The incidence of osteoporosis among Tibet Tibetans ≥50 years old are lower than other nationalities of China in the same age. The participants with high visceral fat mass while low subcutaneous fat mass and muscle mass are more likely to suffer from osteoporosis.

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Measurement of the projection area of Anhui adult’s palm side and the correlation

CHEN Li-wu GAO Guo-zhu

2017, 48 (2): 204-208. doi: 10.16098/j.issn.0529-1356.2017.02.015

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Objective To determine the adult’s palm side projection area and to explore the related influencing factors. Methods The pixel method was used to measure the palm side projection area of 129 living males and females and to measure their hand length, hand width, palm length and palm circumference. Correlation analysis and regression analysis were performed. Stevenson formula was used to calculate body surface area, then to calculate the projection area of palm side accounted for the proportion of body surface area. Results The results of the measurement by applying the method of pixel were more accurate. Hand length and palm circumference were the key indicators to affect the palm side projection area. The projection area of palm side accounted for the proportion of body surface area was less than 1%. Conclusion Hand width can be an alternative of palm circumference，and the estimate formula is Y=8.910X₁+4.242X₂-63.299.

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Research-oriented teaching on the basis of team in stem cell-basics and clinics for graduates

LIU Xing-xia ZHAO Qing PENG Xiao-zhong

2017, 48 (2): 217-221. doi: 10.16098/j.issn.0529-1356.2017.02.017

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Objective To explore the effects of research-oriented teaching on the basis of team in stem cell-basics and clinics for graduates. Methods Forty-one graduates in Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences who participated in the study of stem cell-basics and clinics were consecutively selected. Research-oriented teaching was introduced after traditional lecture-based teaching, and the evaluations were taken at the end, including teaching evaluation, student satisfaction with the study, and assessment of creative ability and project design. Results As shown in the questionnaire survey, the learning interest and the craving for knowledge, the creative ability and the level of project design were effectively improved, the scientific research quality was well trained. Conclusion The research-oriented teaching method can effectively improve graduates’comprehensive ability of self-learning and the creative ability. It is an effective new approach for teaching which is worthy of promotion in graduates’ teaching.

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Anatomic facial makeup of the cranial nerves in human

ZHANG Wei-guang WANG Xu LI Ye ZHANG Yan

2017, 48 (2): 222-224. doi: 10.16098/j.issn.0529-1356.2017.02.018

Abstract ( )

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Objective To investigate the effect of cranial nerves makeup teaching methods on students’ learning of 12 cranial nerves. Methods During the anatomy teaching at Peking University, we had drawn and applied a facial makeup of 12 pairs of cranial nerves in recent three years. Results The teaching method of a facial makeup of 12 pairs of cranial nerves increased the interest, ability of discussion and communication and raised test scores for medical students. Conclusion The ideal teaching effect had been achieved with our teaching method.

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