Table of Content

2019, Volume 50 Issue 2
06 April 2019 Previous Issue    Next Issue
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Anatomy
Cancer Biology
Anthropology
Medical Education
Neurobiology
Cell and Molecules Biology


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Anatomy

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Fenestration and dehiscence in anterior alveolar bone of 52 adolescent patients with skeletal class Ⅱ and high-angle malocclusion assessed with cone-beam computed tomography FENG Jian-kun GAO Jian-hong 2019, 50 (2):  241-244.  doi: 10.16098/j.issn.0529-1356.2019.02.016 Abstract ( )   PDF (2134KB) ( )   Objective To investigate the incidence of dehiscence and fenestration in anterior alveolar bone of 52 adolescent patients with skeletal class Ⅱ and high-angle malocclusion assessed with cone-beam computed tomography, to assist clinical orthodontic treatment. Methods Fifty-two patients aged from 13 to 20 with skeletal class Ⅱ high-angle malocclusion were randomly collected from those who take cone-beam computed tomography（CBCT）examination in Hospital of Stomatology Chengde. Images of all patients were imported to SIDEXIS XG 2.56 software to determine the incidence of the dehiscence and fenestration in anterior alveolar bone. The χ2 test analysis was used to evaluate the incidence differences between sex and jaw as well as tooth site respectively. Results The incidence of the alveolar bone defect was 86.53% with the fenestration incidence(65.38%)and the dehiscence incidence(67.30%) respectively.Eighty-two teeth fenestration and 107 teeth dehiscence were detected out of 416 teeth. The incidence of the fenestration in both jaws was similar(P>0.05) whereas the incidence of the dehiscence in maxillary was lower than the mandibular(P<0.05).Difference incidence is positively related with tooth site(P<0.05) but negatively with sex.Conclusion Dehiscence and fenestration of the adolescent patients with skeletal classⅡand high-angle malocclusion are widely existed in anterior alveolar bone area before the orthodontic treatment. The orthodontics should pay close attention to the alveolar morphology before and during the treatment. References | Related Articles | Metrics

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Construction and application of a web-based three-dimensional organ model visualization system including Chinese anatomical variations WANG Hong-kai ZHANG Nan SUN Xiao-bang ZHANG Yan-jun HUO Li YU Li-juan SHI Hong-cheng ZHANG Bin 2019, 50 (2):  220-226.  doi: 10.16098/j.issn.0529-1356.2019.02.012 Abstract ( )   PDF (9650KB) ( )   Objective Digital three-dimensional (3D) anatomical model have promoted anatomy education and clinical research and treatment. However, most existing digital 3D models of Chinese human are based on standard human subjects which do not include the anatomical variations between different individuals. This limitation hampers the users’ understanding of inter-subject organ morphology variations. Therefore, we constructed a web-based 3D visualization system of human trunk anatomical deformation model to demonstrate these variations. Methods We learned the individual differences of organ morphology from 138 healthy Chinese CT images using the statistical shape model technique, and interactively demonstrated the deformation of anatomical variances through web browser. Results The proposed system realized cross-platform network-based visualization and allowed the adjustment of anatomical deformation through the user’s interactive operation. Conclusion Our web-based system provides novel inter-subject variation information for anatomy education and medical applications. References | Related Articles | Metrics

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Anatomical morphometry of the coracoid and its clinical significance inChniese population based on three-dimensional reconstruction of CT ZENG Sheng-qiang MA Li ZHANG Lei WANG Guo-you FU Shi-jie GUO Xiao-guang DENG Kai BAO Ding-su 2019, 50 (2):  227-231.  doi: 10.16098/j.issn.0529.1356.2019.02.013 Abstract ( )   PDF (2172KB) ( )   Objective To study the anatomical morphological features of coracoid process by 3D reconstruction of scapular CT, and to provide anatomic basis for clinical diagnosis and treatment of related diseases. Methods The imaging data of 256 patients with CT three-dimensional reconstruction of scapula were selected and the coracoid process related anatomical data were measured and compared and analyzed in the Hospital of Traditional Chinese Medicine Affiliated to Southwest Medical University. Results The anatomical data of 256 cases of coracoid process reconstruction model were analyzed. The width of the tip of the coracoid process was (14.31±2.71) mm and the thickness of the apex was (8.79±1.44)mm. The width of the folded part was (22.87±2.82) mm, the thickness of the folded part was (14.68±2.39) mm, the width of the base was (26.29±3.05) mm, the length of the base was (10.71±3.01) mm, the length of the tip to the base was (19.36±2.35) mm. There were significant differences in the thickness of the coracoid tip, the width of the reentrant part and the length from the reentrant part to the base of the coracoid process (P<0.05), but there was no significant difference in other anatomical morphology of the coracoid process (P>0.05). Conclusion Three-dimensional CT reconstruction can provide anatomical parameters of coracoid process and provide anatomic basis for clinical diagnosis and treatment. References | Related Articles | Metrics

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Influences of nonplanarity on the measurements of the posterior semicircular canal angles YANG Xiao-kai ZHENG Yan-yan CHEN Xiao-su 2019, 50 (2):  232-235.  doi: 10.16098/j.issn.0529-1356.2019.02.14 Abstract ( )   PDF (3772KB) ( )   Objective To measure the angles among the posterior semicircular canal, and to observe the influences of nonplanarity on the measurements. Methods With 3D Slicer,we directly measured angles among the left and right posterior semicircular canals from MRI volume data of 20 patients which underwent MRI scaning with 3D-CISS sequence,and then calculated angles among the posterior semicircular canals and sagittal head plane. Results The angle between the left and right posterior semicircular canals was 190.5°±8.9° between 73° and 108°,and the angle among the posterior semicircular canals and sagittal head plane was 45.3°±4.4° between 36.5°and 54°. Conclusion Whether the plane including or excluding the common crus will result in differences of up to 9°,therefore, this study is consistent with previous studies but this method is more intuitive and convenient. References | Related Articles | Metrics

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Measurement of orbital aperture area and the relationship with aging of both genders in Zhejiang  PAN Lei FENG Wei ZHAO Ye JIN Ting-ting LI Jing-yu WU Su-fan 2019, 50 (2):  236-240.  doi: 10.16098/j.issn.0529-1356.2019.02.015 Abstract ( )   PDF (6298KB) ( )   Objective To study the significance of parameters of bony orbit in different age groups of Chinese population in both genders，providing anatomical evidence and strategy for clinical rejuvenation. Methods The data were obtained from 114 adults from Zhejiang province who underwent computed tomographic examinations followed by 3D reconstruction with volume rendering. They were divided into 3 groups: young, mid-age and old-age groups. The measurements of orbital aperture area (S) that including four subareas (S1, S2, S3, S4), up-to-bottom diagonal (L1), and mediumtolateral diagonal (L2) were obtained and calculated by Adobe Photoshop software. The comparison was made among age and gender groups. Results There was significant increasing of up-to-bottom diagonals (L1) with aging in both genders (P<0.05), and a sexual difference appeared in mid-age group. The medium-to-lateral diagonals (L2) for male subjects were longer than female in all age groups(P<0.05). But L2 did not change with aging. The total orbital aperture area of male subjects had no obvious change, while the subarea S1, S2, and S3 significantly increased and S4 decreased with aging. The orbital aperture area of female subjects rose from young to old-age group and the most augmented parts were S3 and S4. There was significant difference between male and female subjects in mid-age group. Conclusion The bony orbital aperture area changed with aging, but the changing pattern varied in male and female subjects. The study and analysis of the morphological change of the orbital aperture is significant to our clinical treatment in cosmetic rejuvenation. References | Related Articles | Metrics

Cancer Biology

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MicroRNA-140-5p promotes cell proliferation via protein kinase C isoform ε in renal cell carcinoma  LIU Wei-hong YU Xiao REN Yu LIU Kai-tai LIN Chen WANG Ping 2019, 50 (2):  185-191.  doi: 10.16098/j.issn.0529-1356.2019.02.008 Abstract ( )   PDF (2571KB) ( )   Objective To explore the role of micro RNA-140-5p(miR-140-5p) in renal cell carcinoma (RCC) tumorigenesis. Methods The expression of miR-140-5p was determined by Real-time PCR in RCC (Caki-1, ACHN, OS-RC-2 and 786-O), and compared with kidney cortex proximal tubule cell (HK-2). After Caki-1 and 786-O cells were transfected with miR-140-5p inhibitor or mimic, the cell proliferation, cell cycle and related proteins were evaluated by MTS assay, flow cytometry and Western blotting. Results Caki-1 and 786-O showed the highest and lowest expression of miR-140-5p respectively. After the level of miR-140-5p in Caki-1 cells was decreased, cell proliferation was inhibited and cell number in G0/G1 phase was increased significantly, along with the reduction of cyclin D, E and CDK2, 4. Furthermore, the increasing level of miR-140-5p in 786-O cells promoted cell proliferation in vitro and tumorigenesis in vivo.Moreover, microarray demonstrated that protein kinase C isoform ε(PKCε), a markedly enhanced gene in RCC cells with the changed level of miR-140-5p. The combination treatment with down-regulation of PKCε, inhibitor/mimic induced changes of cell proliferation and cell cycle arrests were marked reversed in Caki-1 and 786-O cells.Conclusion These result indicate that miR-140-5p may promote cell proliferation via PKCεin RCC. References | Related Articles | Metrics

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Up-regulation of serine protease inhibitor Kazal type 1 expression in human lung adenocarcinoma and promotion of proliferation of cancer cells  LU Chang-chang HUANG Yan-yan XU Li-yun HE Jian-ying QIU lei LE Han-bo 2019, 50 (2):  192-200.  doi: 10.16098/j.issn.0529-1356.2019.02.009 Abstract ( )   PDF (10472KB) ( )   Objective To explore the relationship between serine protease inhibitor Kazal type 1(SPINK1) expression and survival time of lung adenocarcinoma (LAC) and its effect on the growth of LAC cells. Methods The SPINK1 expressions in 285 cases of lung adenocarcinoma tissues and adjacent tissues were detected by Real-time PCR and immunohistochemical staining. The relationship between SPINK1 expression and clinicopathological features and prognosis of lung adenocarcinoma was analyzed. Human LAC cells were treated with lentivirus vector with SPINK1 or SPINK1 siRNA. Green fluorescent protein(GFP) expression was observed under microscope. SPINK1 expression was detected by enzyme-linked immunosorbent assay (ELISA) or Real-time PCR. The growth of LAC cells was evaluated by cell count and colony formation assay. Results The levels of SPINK1 mRNA and protein level in lung adenocarcinoma were up-regulated (P<0.0001). The overall survival time in the group of SPINK1 high expression was shorter than that of SPINK1 low expression (50.0 months vs 64.5 months, P<0.001). In SPINK1 overexpression group, SPINK1 protein was up-regulated in the cultures of LAC cells (P<0.05), and the ability of proliferation and colony formation was up-regulated (P<0.05). In SPINK1 knockdown group, SPINK1 mRNA was down-regulated in LAC cells (P<0.05), and the ability of proliferation was decreased (P<0.05). Conclusion High expression of SPINK1 in lung adenocarcinoma suggests poor prognosis. SPINK1 promotes the proliferation of LAC cells. References | Related Articles | Metrics

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Troubles and answers of identity authentication of human tumor cell lines established in China BIAN Xiao-cui LIU Yu-qin YANG Zhen-li FENG Hai-liang 2019, 50 (2):  201-210.  doi: 10.16098/j.issn.0529-1356.2019.02.010 Abstract ( )   PDF (4099KB) ( )   Objective To authenticate the identity of human tumor cell lines established in China and make sure the authenticated cell lines will be used in science and technology research in China. Methods We collected and extracted genomic DNA of those tumor cells, amplify short tandem repeat (STR) loci by PCR and performed electrophoretic analysis to get STR profiles (as the DNA finger print of the cell line). Comparing their STR profiles against international STR database and those of their donor tissues, crosscontaminated cell lines will be excluded from the service list. PCR detection of human papilloma virus type 18（HPV-18）fragment were performed to double check HeLa crosscontaminated cell lines. Suspected cells contaminated by T leukemia cells were checked by detection characteristics of T cells, such as expression of some surface markers. Results Altogether 46 samples of 37 human tumor cell lines were collected from different labs. All 6 cell lines established in our center showed identical STR profile with their original tissue or primary culture. Among the tumor cell lines established by others 8 cell lines showed unique STR profile, indicating authentic. The other 23 (62.2%) of them showed identical STR with HeLa or with high evaluation value (EV) value, HPV fragment were found in their genomic DNA indicating replacement by or hybrid with HeLa. Some tumor cell lines established in China were cross-contaminated by human colorectal cancer cell line HCT-8, cervical squamous cancer line SiHa, T leukemia cell line CCRF-CEM and a rat cell line. Conclusion A large proportion of human tumor cell lines established by Chinese scholars were cross-contaminated. So scientists must first do authenticate these cell lines before experiments or obtain cell lines from China Infra-structure of Cell Line Resource. Furthermore, when trying to establish cell lines, preserve original tissue/DNA for future authentication. References | Related Articles | Metrics

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Effect and molecular mechanism of trefoil factor family 3 on the cell cycle of thyroid papillary carcinoma cell DAI Jin LIN Xu GUO Meng-yao ZHANG Jing ZHANG Wen-jing XUE Gang WU Jing-fang 2019, 50 (2):  211-219.  doi: 10.16098/j.issn.0529-1356.2019.02.011 Abstract ( )   PDF (12940KB) ( )   Objective To investigate the effects of trefoil factor family 3(TFF3) gene silencing on proliferation and cell cycle of papillary thyroid carcinoma TPC-1,BCPAP cell lines and related molecular mechanisms. Methods To obtain TPC-1 and BCPAP stable cell lines of thyroid papillary carcinoma by packaging TFF3 shRNA lentiviral vectors and virus infection;Growth curve and colony formation assay were used to detecte cell proliferation after TFF3 silencing.Flow cytometry was used to detect the effects of TFF3 gene on cell cycle of TPC-1 and BCPAP cell; Real-time PCR was used to investigate the mRNA levels of P27,P21,cyclin D1 and cyclin-dependent kinase(CDK)4,in TPC-1 and BCPAP cell. The protein expression levels of P27,P21,cyclin D1 and CDK4,protein kinase B(Akt) and phosphorylated protein kinase B(pAkt) were detected by Western blotting and immunocytochemistry. Results shTFF3-TPC-1 and shTFF3-BCPAP stable cell lines were obtained by TFF3 shRNA lentiviral packaging and virus infection. Growth curve and colony formation assay result showed that cell viability was decreased after silencing TFF3.The result of flow cytometry showed that the proportion of cells in the G1 phase was significantly increased (*P<0.05), however the proportion of cells in the S and G2 phase was significantly decreased in TFF3 slienced group compared with the control group (*P<0.05)；The expression of P27 and P21 mRNA and protein in TPC-1 and BCPAP cells was significantly up-regulated(*P<0.05),while the expression of cyclin D1 and CDK4 mRNA and protein was decreased in group TFF3 silenced (*P<0.05).There was no significant difference in the content of Akt protein in the four groups of cells, but the expression of pAkt was weakened after TFF3 silenced;Immunocytochemical staining showed that cyclin D1 positive protein expression was located in the nucleus of cancer cells, P27 and P21 proteins lo ated in cytoplasm and nucleus,and after silencing TFF3, the positive signals were upregulated in the nucleus, but cytoplasmic positive expression was decreased. Conclusion Silencing TFF3 can significantly prolong the cell cycle and inhibit cell proliferation，which might be related to the inhibition of phosphatidylinosital 3-kinase and protein kinase B(PI3K/Akt) pathway-related protein expression. References | Related Articles | Metrics

Anthropology

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Morphological characteristics of lips of female in Lhasa Tibetan,Kham Tibetan and Amdo Tibetan   ZHONG Hua WANG Fei LI Xiao-jing LI Guang YU Li-xia CI Ren BAIMA Duo-ji CHU Duo LUO Sang LIU Ya-li DING Ya-li 2019, 50 (2):  245-248.  doi: 10.16098/j.issn.0529-1356.2019.02.017 Abstract ( )   PDF (779KB) ( )   Objective To investigate the lip index and mouth breadth of the native Tibetan nationality female. Methods A total of 542 Tibetan nationality people aged 18-23 years old in Lhasa were selected to measure the mouth breadth,lip height and lip index. Results The mean of the Tibetan nationality female’s lip height was(18.01±4.29)mm, mouth breadth was(49.40±5.55)mm, and lip index was (36.60±8.21)mm. The mean values of lip height and lip index of the Kham Tibetan were significantly different from those of the Lhasa Tibetan and the Amdo Tibetan (P<0.01). There was significant difference in the mean mouth breadth between Lhasa Tibetan and the Amdo Tibetan (P<0.01).There was a significant negative correlation between lip height and age(r=-0.124,P<0.01),there was a significant negative correlation between lip index and age(r=-0.155,P<0.01),There was no statistically significant correlation between the width of the mouth and the age（P＞0.05）. Conclusion Kham Tibetan has a higher lip height, and the Amdo Tibetan has a wider mouth breadth. Tibetan nationality female lip index indicates the normal lips group as the main (69.2%), and wide group with mouth breadth as the main (73.6%). References | Related Articles | Metrics

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Sex differences of manubrium in Macaca mulatta and its correlation with the length of clavicle and femur  ZHAO Xiao-jin HAN Xiao-fan WANG Feng-chan 2019, 50 (2):  249-253.  doi: 10.16098/j.issn.0529-1356.2019.02.018 Abstract ( )   PDF (914KB) ( )   Objective To evaluate the sexual dimorphism in the sternum, and further to investigate whether a correlation exists between the sternal variables and the length of the clavicle and femur. Methods We examined intact sterna, clavicle and thighbone from 45 Macaca mulatta (13 males and 32 females). Six linear variables, including four sternal variables, length of clavicle and length of femur, were measured. Statistical analysis was done using SPSS 22.0 package, and t-test was used to determine the difference between sexes. A receiver-operator characteristic (ROC) curve analysis was conducted in order to identify the sensitivity and specificity of gender identification of each variable. The sex discrimination rate of each variable was derived using the discriminant analysis. Results The sexual differences of sternal variables were statistically significant (P<0.05). The correct discrimination rate was higher in anterior width of sternum (84.4%) and relatively lower in posterior width of sternum (66.7%). The correlation was higher between sternal variables and length of clavicle and weaker between sternal variables and length of femur. The error was larger when estimating stature using manubrium. Conclusion The manubrium of Macaca mulatta can be used for estimation of sex. The principal factors influencing sternum morphology may be locomotor and habit behavior. References | Related Articles | Metrics

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Correlation between the dizziness of the middle and young people and the abnormal curvature of the cervical spine WANG Ying TAN Jun 2019, 50 (2):  254-258.  doi: 10.16098/j.issn.0529-1356.2019.02.019 Abstract ( )   PDF (1156KB) ( )   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, collecting the young patients with vertigo from the Department of Neurology of the Third Affiliated Hospital of Xinxiang. There were 78 cases, of which were 32 males and 46 females.Based on whether the cervical curvature change, dividing the patients into normal cervical curvature group (n=22)and abnormal cervical curvature group(n=56), analysis of the clinical characteristics between the two groups we carried out, to investigate the correlation between cervical curvature change and the different type of dizziness. Results The ABCD2 scores and dizziness clinical symptoms were significantly higher than those in the normal cervical flexure group, and the difference was statistically significant (P<0.05). There was a high positive correlation between the abnormal curvature of the cervical spine and the different dizziness types (P<0.01). Conclusion With the prolonging of sick time, the change of cervical lordosis increases, clinical symptoms of dizziness in youth is more and more serious.  References | Related Articles | Metrics

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Body density of the Mongolian in China  LI Yong-lan ZHENG Lian-bin 2019, 50 (2):  259-263.  doi: 10.16098/j.issn.0529-1356.2019.02.020 Abstract ( )   PDF (774KB) ( )   Objective To study the body density of thirteen Mongolian groups, to compare their gender and ethnic differences, and to investigate the correlation between the body density and longitude, latitude, annual average temperature and age. Methods The triceps skinfold and the subscapular skinfold were measured. Jinji, Changling formula was used to calculate the body density of thirteen Mongolian groups. Results In the thirteen Mongolian groups, the body density of Yunnan Mongolian was the largest, and that of Ejina Torghut Mongolian was the least. The differences of the Mongolian body density among ethnic groups, gender, age groups and 4 regions were statistically significant. The body density of Mongolian had a significant negative correlation with longitude and age. Conclusion The body density of the southern Mongolian is greater than that of the northern Mongolian, and the body density of the western Mongolian is greater than that of the eastern Mongolian. References | Related Articles | Metrics

Medical Education

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Teaching model design and experimental study of intelligent scalpel holding training  YU Meng ZHANG Wei-tao ZHANG Peng LI Hao-xu 2019, 50 (2):  264-268.  doi: 10.16098/j.issn.0529-1356.2019.02.021 Abstract ( )   PDF (2465KB) ( )   Objective To effectively enhance the scalpel holding training, reduce the environmental requirements for training, and change the current experimental-teaching method . Methods We proposed a design method for intelligent scalpel holding training model, and corrected the design and experiment of version 2. Results The designed training model not only simulated anatomical work scene, but also intelligently recognized the hand-type of holding scalpel and the scalpel knife action. Concretely, the proposed model was able to apperceive the scalpel holding angle, force, insertion and evulsion, the direction, speed and distance of scalpel holding training. By using the four scalpel holding training plans, the proposed model completed the whole experimental teaching process, including scalpel holding training, interactive voice guidance and evaluation of the training result. Conclusion The experimental result shows that the teaching guidance model can greatly enhance the scalpel holding training teaching effect and efficiency, and is a worthy promotion of medical experimental teaching aids and training equipment. References | Related Articles | Metrics

Neurobiology

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Effect of resveratrol on activation of microglia cell line N9 after oxygen-glucose deprivation/reoxygenation injury in vitro LIAO Hong-yan LIU Jie LIU Jing GUO Shuang YANG Qin 2019, 50 (2):  137-144.  doi: 10.16098/j.issn.0529-1356.2019.02.001 Abstract ( )   PDF (4464KB) ( )   Objective To investigate the effect of resveratrol on the activation of microglia cell line N9 after oxygen-glucose deprivation/reoxygenation (OGD/R) injury in vitro.Methods N9 microglias were cultured under oxygen-glucose deprivation for 150 minutes and reoxygenation for 24 hours. The experiments were divided into normal (Nor), control (Ctrl) and resveratrol pretreatment group (Res). Cell viability was determined by cell counting kit-8(CCK-8) assay. The percentage of apoptosis was measured by flow cytometry. Expression of ionized calcium binding adaptor molecule-1(Iba1), CD11b, Caspase-3, Bax，interleukin-10 (IL-10), tumor necrosis factor α(TNF-α) and IL-1β proteins were detected by immunofluorescence. The levels of IL-10, TNF-α and IL-1βprotein in the supernatant were measured by ELISA. Results CCK-8 assay showed that the viability of microglia in the control and 1,5,20,40,80 μmol/L resveratrol groups significantly decreased compared with the normal group after OGD/R injury (P＜0.05，n=3), and significantly enhanced in the 5, 20, and 40 μmol/L resveratrol group than those in the control group, the peak was 20 μmol/L resveratrol group (P＜0.05,n=3). Immunofluorescence, Western blotting and ELISA or flow cytometry method showed that expressions or levels of CD11b, Iba1, Caspase3, Bax, IL-10, TNF-α, IL-1β proteins and the percent of apoptosis significantly increased in the control and the resveratrol group after OGD/R injury (P<0.05, n=3). However, except the expression or level of IL-10 protein was higher, the rests were lower in the resveratrol group than those of the control group (P<0.05, n=3). Conclusion Resveratrol pretreatment can inhibit the activation and apoptosis of microglia, reduce inflammation after OGD/R. References | Related Articles | Metrics

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Neuroprotective effect of γ-secretase inhibitors on neural stem cells after cerebral ischemia in rats LIU Zong-xiu ZHANG Zi-wei ZHOU Xiao-hong CHENG Yuan GAO Wei-juan 2019, 50 (2):  145-151.  doi: 10.16098/j.issn.0529-1356.2019.02.002 Abstract ( )   PDF (8604KB) ( )   Objective To investigate the neuroprotective effects of γ-secretase inhibitors N-［N-(3,5-difluorohenacetyl-L-alanyl)]-S-phenylglycinet-butylester(DAPT) on neural stem cells (NSCs) after cerebral ischemia in rats. Methods Thirty-three SD rats were randomly divided into three groups: model group, neural stem cell transplantation group (transplantation group) and DAPT+ transplantation group. In the other sham operation group, only the exposed blood vessels were separated under anesthesia. The neurobehavioral changes of rats in each group were observed 7 days after transplantation. The volume of cerebral infarction was observed by TTC staining. The pathological changes of brain tissue were observed by Nissl staining. The expression of BrdU/neuronal nuclei(NeuN) positive cells was detected by immunofluorescence double labeling. Results The neurological function score of the sham operation group was zero, no cerebral infarction, cell morphology was intact, Nissl bodies were abundant, and BrdU/NeuN positive cells were negative. Compared with the sham operation group, the neurological scores of the model group increased. There were obvious symptoms of neurological deficits, obvious cerebral infarction, disordered arrangement of nerve cells, nuclear pyknosis, rare Nissl bodies, and a significant decrease in the number of surviving neurons (P<0.05), with a small amount of BrdU/NeuN positive cells expression (P<0.05); Compared with the model group, the transplant group and the DAPT+ transplantation group had different degrees of neurological function score reduction, the cerebral infarction volume decreased, and the number of nerve cell survival increased (P<0.05), BrdU/NeuN expression of positive cells increased (P<0.05), and the indexes of DAPT+ transplantation group recovered most obviously. Conclusion DAPT can promote the differentiation of transplanted NSCs into neurons, and has neuroprotective effects on neural stem cells after cerebral ischemia. References | Related Articles | Metrics

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Mechanisms of pain hypersensitivity in a mouse model of chronic compression of dorsal root ganglia  TAO Jin WANG Tao ZHU Jie MA Chao 2019, 50 (2):  152-157.  doi: 10.16098/j.issn.0529-1356.2019.02.003 Abstract ( )   PDF (3191KB) ( )   Objective To investigate the mechanisms of pain hypersensitivity to lowdose capsaicin in a mouse model of chronic compression of dorsal root ganglion (CCD). Methods Chronic compression of L4 DRG was performed in mice by inserting an L-shaped stainless steel rod into the L4 intervertebral foramina. Different doses of capsaicin（0.01, 0.1, 1 g/L）1 μl were injected into the skin on the calf area and behavior responses were videotaped on pre-CCD 1 days and post-CCD 1, 3, 5, 7 days. The optimal concentration that led to a significant difference after CCD was determined and was used in the following in-vivoDRG imaging studies. Immunofluorescent staining was conducted to evaluate the expression of transient receptor potential vanilloid 1 (TRPV1) in DRG from na?ve and CCD mice. Results Behavioral tests showed that 0.1 g/L capsaicin elicits a significant difference in pain-like behaviors after CCD（n=8; post-CCD 1 day, P<0.01; post-CCD 5, 7 days P<0.01; post-CCD 3 days, P> 0.05. In vivocalcium imaging showed an enhanced number of activated DRG neurons to the injection of capsaicin in CCD mice, which was 75 in total of 398(n=4) for control mice and 169 in total of 382(n=6) (P<0.01). According to immunofluorescent staining results, there were 148 TRPV1+ neurons in total 653 counted neurons (n=10) for control mice and 237 TRPV1+ neurons in total of 611 neurons (n=6) for CCD mice (P<0.01). Conclusion Chronically compressed DRG neurons show upregulated TRPV1 receptor and enhanced responses to low-dose capsaicin, that produce pain hypersensitivity in the CCD mice. References | Related Articles | Metrics

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Effects of cystatin C pretreatment on expression of heat shock protein 70，microtubule-associated protein light chain-3 and ultrastructure in rats with cerebral cortex after ischemia and reperfusion CHEN Meng DU Chao TIAN Huan-na WANG Ai-le XIA Xiao-xiao 2019, 50 (2):  158-165.  doi: 10.16098/j.issn.0529-1356.2019.02.004 Abstract ( )   PDF (15232KB) ( )   Objective To study the effect of different concentrations of cystatin C (Cys C) pretreatment on apoptosis related proteins, the expression of heat shock protein 70(HSP70) and autophagy related proteins, the expression of microtubule-associated protein light chain-3(LC3) in rats following cerebral ischemia reperfusion injury. Methods The healthy sixty male SD rats were randomly divided into five groups: the sham operation group(sham), the cerebral ischemia reperfusion injury group(IR) and low concentration of Cys C group(Cys C1), middle concentration of Cys C group(Cys C2) and high concentration of Cys C group(Cys C3), 12 rats in each group. The model of cerebral ischemia reperfusion injury was established by a suture method. After ischemia for 2 hours and reperfusion 24 hours, the modified neurological severity scores were detected. Using Western blotting method to detect the expression of HSP70, LC3-Ⅱ/LC3-Ⅰprotein. The number of HSP70 and LC3 positive cells were detected by immunohistochemical SP method . The average absorbance of autophagy related protein LC3/neuronal nuclei (NeuN) in corticalneurons was detected by immunofluorescence double labeling assay. The structural changes of the neurons were observed under transmission electron microscope. Results Compared with the IR group, the mNSS score significantly reduced(P<0.05), and upregulated the expression of HSP70 in Cys C1, Cys C2 group(P<0.01). While the mNSS score obviously increased in Cys C3 group, there was no significantly different(P>0.05) between Cys C3 group and IR group; the expression of HSP70 decreased in the Cys C3 group(P<0.01); the expression of LC3-Ⅱ/LC3-Ⅰprotein in each concentration cystatin C group increased gradually(P<0.01). The average absorbance of LC3/NeuN in the each concentration cystatin C group increased gradually (P<0.01). The results of transmission electron microscope showed that the ultrastructure pathological changes of cerebral cortex neurons could alleviated in Cys C1 and Cys C2 group, there were many obvious autophagic lysosomes. While the ultrastructural damage of cerebral cortex neurons in Cys C3 group was very serious. Conclusion Within a certain concentration range, Cyst C has a protective effects on ischemia reperfusion injury neurons. Otherwise, excessive autophagy maybe leads to autophagic death. References | Related Articles | Metrics

Cell and Molecules Biology

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Knockdown of calcium/calmodulin-dependent protein kinaseⅣ reduce the immune characteristic of C2C12 under stimulation of interferon-γ LI Jun-hua MA Yong-neng GU Rui-cai LIAO Hua 2019, 50 (2):  166-172.  doi: 10.16098/j.issn.0529-1356.2019.02.005 Abstract ( )   PDF (9420KB) ( )   Objective Whether calcium/calmodulin-dependent protein kinase Ⅳ (CaMKⅣ) plays a role in regulating immunologic features of muscle cells in inflammatory environment, remains mostly unknown. In this study, we investigated the influence of CaMKⅣ on the immunological characteristics of myoblasts and myotubes received interferon(IFN)-γ stimulation. Methods To investigate the effects of CaMKⅣ on immune characteristics of C2C12 myoblasts and differentiated myotubes, which are firstly knocked down endogenous CaMKⅣ gene and then treated with IFN-γ. Real-time PCR and Western blotting were performed to analyze the expression of CaMKⅣ. Western blotting and immunofluorescence were performed to analyze the expression of major histocompatibility complex(MHC) class-Ⅰ,MHC class-Ⅱ,Toll-like receptor3(TLR3). The expression of interleukin(IL)-1β,IL-6,tumor necrosis factor(TNF)-α,macrophage inflamator protein(MIP)-1α and monocyte chemoattractant protein(MCP)-1 were measured by Real-time PCR. Results Under IFN-γ induced pro-inflammatory milieu, MHC-Ⅰ molecule H-2Kb, MHC-Ⅱ molecule H2-Ea, TLR3 significantly up-regulated in myoblasts and in differentiated myotubes. In striking contrast, CaMKⅣ inhibition in myoblasts and myotubes led to expression suppression of the above molecules. As well, gene levels of pro-inflammatory IL-1β, IL-6, TNF-α, MIP-1α and MCP-1 in C2C12 cells (especially in myotubes) were markedly down-regulated by CaMKⅣ knocking off. Conclusion Knockdown of CaMKⅣ gene can effectively inhibit the expression of IFN-γ-induced immune molecules, suggesting that CaMKⅣ is involved in regulating the immunological properties of muscle cells. References | Related Articles | Metrics

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Expression of anoctamin 1 in the process of myocardial fibrosis  TIAN Xiang-qin TAN Zhao-yang LI Xin-zhi MA Ke-tao LI Li SI Jun-qiang﹡ 2019, 50 (2):  173-178.  doi: 10.16098/j.issn.0529.1356.2019.02.006 Abstract ( )   PDF (8400KB) ( )   Objective To clarify the expression feature of anoctamin 1(ANO1) in the process of cardiac fibrosis and seek new targets for preventing it. Methods The myocardial infarction (MI) rats were prepared by coronary artery ligation. The left ventricular from MI group and the sham operation group were taken at one week (1 week) after MI. The changes of ANO1 expression in the two groups were detected by immunohistochemical staining and immunofluorescence double labeling. The expression of ANO1 in cardiac fibroblasts (CFs) was detected by immunofluorescence labeling, Real-time PCR and Western blotting. Results ANO1 exhibited high expression in MI and co-expressed with α-smooth muscle actin(α-SMA), and it was clearly expressed in the nucleus membrane and cytoplasm of CFs. The expression intensity of ANO1 was coincident with α-SMA. Furthermore, ANO1 expression increased significantly in the CFs after 48 hours culture comparing with the new isolated CFs. Conclusion The expression of ANO1 enhances markedly during the transformation of CFs into myocardial fibroblasts, suggesting that ANO1 may play an important role in the process of cardiac fibrosis. References | Related Articles | Metrics

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A preliminary study on the protective effect and mechanism of paraoxonase 3 gene overexpression on hepatic injury in acutely poisoned mice WU Yang LIU Ying LIU Ji-tao YIN De-feng 2019, 50 (2):  179-184.  doi: 10.16098/j.issn.0529-1356.2019.02.007 Abstract ( )   PDF (4154KB) ( )   Objective To investigate the protective effect and mechanism of oxidative sulfasalase 3 on acute poisoned liver injury. Methods Forty Balb/c mice were randomly divided into 4 groups: normal control (NC) group, dichlorvos (DDVT) control group, green fluorescent protein lentivirus (Lv-GFP) group, and recombinant paraoxonase 3(PON3) lentivirus (Lv-PON3) group, 10 in each group.Mice in Lv-GFP and Lv-PON3 group were injected with 2×107 TU of Lv-GFP and Lv-PON3 lentiviruses via the tail vein respectively. After 3 days, they were intraperitoneally injected with a solution of dichlorvos (DDVT) 9 mg/kg.The same dose of DDVT was injected intraperitoneally into the DDVT group, and the same dose of saline was injected into the NC group.In each group, 10 mice were killed by anesthesia 12 hours after DDVT exposure to take liver tissue. ELISA was used to detect serum acetylcholinesterase(AChE), liver function indexes alamine aminotransferase(ALT) and aspartate aminotransferase(AST) and oxidative stress indexes malondialdehyde(MDA), catalase(CAT), superoxide dismutase(SOD) and glutathione(GSH) in liver tissues of each group. HE staining was used to observe the morphological changes of liver tissues. Results Compared with the NC group, the levels of MDA, ALT, AST, hemoglobin oxygenase 1(HO-1) and Nrf2 mRNA expression were significantly increased in DDVT group and Lv-GFP group, and AChE, CAT, SOD, GSH levels and Kelch-like epichlorohydrin-associated protein 1(Keap1)mRNA were significantly decreased (P<0.05). Compared with the DDVT group, the levels of MDA, ALT, AST and Keap1mRNA expression in liver tissue were significantly decreased in Lv-PON3 group, and AChE, CAT, SOD, GSH levels and HO-1 and Nrf2 mRNA expression were significantly increased (P<0.05). The structure of hepatocytes in the normal control group was clear, no hepatocyte necrosis and fat lesions were seen. Pathological changes such as hepatocyte necrosis and fatty degeneration under light microscope in DDVT and Lv-GFP groups, the number of hepatocyte necrosis and steatosis cells in the liver lesions of the Lv-PON3 group were reduced. Conclusion PON3 can alleviate oxidative stress and improve liver function through Keap1-Nrf2/HO-1 pathway, which has a certain protective effect on liver damage induced by acute poisoning of dichlorvos in mice. References | Related Articles | Metrics