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    Cancer Biology
     Effect of Survivin shRNA on c-myc and nuclear factor-κB expression in esophageal cells
    FENG Ming Myire?Tuerhong DONG Juan-juan Msa?Aerken LI Xiu-mei
    2019, 50 (1):  72-76.  doi: 10.16098/j.issn.0529-1356.2019.01.012
    Abstract ( )   PDF (2468KB) ( )  

    Objective To study the effect of Survivin shRNA interference on c-myc and NF-κB expression levels in esophageal cells ECA109. Methods Survivin shRNA interference technology was applied to liposomes transfection esophageal ECA109 cells. Half quantitative RT-PCR and Western blotting method were applied to detect genes expression of c-myc and NF-κB gene. Flow cytometry was used to detect the proliferation and apoptosis of cell lines index changes. Results 1. After Survivin shRNA transfected esophageal ECA109 cells, Survivin mRNA and protein expression obviously down-regulated, and c-myc mRNA and protein expression also down-regulated; 2. After Survivin down-regulated, NF-κB mRNA expression had no obvious change, however the phosphorylated proteins level of NF-κB was reduced, whose upstream factor nuclear factor kappa B inhibitor kinase predominate(IKK) alpha, IKK beta mRNA express was cut; 3. Esophageal cell ECA109 apoptosis increased significantly. Cells ratio increased in G2 phase but decreased in S phase. The cell cycle blocked. Conclusion Survivin is effective on adjusting c-myc gene expression on mRNA and protein level. Survivin regulates NF-κB activation by down-regulation of IKK alpha, IKK beta mRNA expression.

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    Expression and mechanism of microRNA-940 in breast cancer
    XUE Shi-hang LU Zhen-yi ZHANG Tong-cheng ZHU Cong-lun
    2019, 50 (1):  56-62.  doi: 10.16098/j.issn.0529-1356.2019.01.010
    Abstract ( )   PDF (3033KB) ( )  

    Objective To explore microRNA-940(miR-940) expression in breast cancer tissues and cells, as well as its effect on breast cancer cell proliferation, invasion and migration capacities and the related molecular mechanism. Methods miR-940 expression in breast cancer tissues, para-carcinoma tissues from 78 patients undergoing surgical resection in our hospital from January 2016 to January 2017, and human breast cancer cell lines MCF-7, SK-BR-3, MDA-MB-231 and BT-549, as well as that in normal human breast cell line MCF-10 A, were detected using Real-time PCR. miR-940 expression in breast cancer cell line MDA-MB-231 was up-regulated through transfection with miR-940 mimics using liposome LipofectaminsTM 2000. Changes in cell proliferation capacity were detected using CCK-8 assay, while those in cell invasion and migration capacities were detected using transwell assay. The potential target gene of miR-940 was predicted by biologic information method , binding of miR-940 with the 3’UTR of CXC chemokine receptor 2 (CXCR2) was detected using dual luciferase reporter assay, and the effect of miR-940 on CXCR2 protein expression was detected using Western blotting. Results miR-940 expression in breast cancer tissues and cells was remarkably down-regulated (P<0.01), and miR-940 expression was closely correlated with TNM stage and lymph node metastasis (P<0.01). Up-regulating miR-940 expression in MDA-MB-231 cells would lead to markedly decreased cell proliferation capacity (P<0.01), as well as notably lowered cell invasion and migration capacities (P<0.01). Dual luciferase reporter assay indicated that miR-940 might bind with specific sequences in the 3’UTR of CXCR2, thus evidently suppressing luciferase activity (P<0.01); meanwhile, up-regulating miR-940 would result in apparently reduced CXCR2 protein expression in cells (P<0.01). Conclusion miR-940 expression is down-regulated in breast cancer, which can target CXCR2 to suppress breast cancer cell proliferation, invasion and migration.

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    Cancer Biology
    Features of CT reconstruction images and expressions of epithelia-mesenchymal transition related genes in lung adenocarcinoma with micropapillary pattern
    ZHOU Ji-hang XU Li-yun LU Chang-chang HUANG Yan-yan WANG Xin-chen LE Han-bo
    2019, 50 (1):  63-71.  doi: 10.16098/j.issn.0529-1356.2019.01.011
    Abstract ( )   PDF (7774KB) ( )  

    Objective To investigate the features of CT reconstruction imags, expressions of genes associated with epithelial-mesenchymal transition(EMT) in lung adenocarcinoma with micropapillary pattern (MPP) and the relationships between them. Methods Thirty-seven cases of invasive lung invasive adenocarcinoma (IAC) with MPP, 80 cases without MPP and their corresponding para-cancerous tissues were selected. The clinicopathological features and features of CT reconstruction images were compared. Real-time PCR and Western blotting were used to detect the expression of EMT markers. Results In MPP group, the proportions of lymph node metastasis, spicule, pleural indentation, solid type and lobulation were higher(P<0.001).In MPP group, the expressions of E-cadherin, β-catenin were decreased(P<0.05), and the expressions of N-cadherin, vimentin,Snail and transforming growth factor(TGF)-β were increased(P<0.05). In spicule group, the percentage of decreased expression of E-cadherin and β-catenin was higer, while the percentage of increased expression of vimentin was higher(P<0.05). In the group of solid type, the percentage of decreased expression of E-cadherin was higher, while the percentages of increased expression of N-cadherin and vimentin were higher(P<0.05). Conclusion Lung invasive adenocarcinoma has characteristics of CT reconstruction images and EMT changes. CT reconstruction images are associated with the expression of the related molecules associated of EMT.

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    Anthropology
    Physical anthropological characteristics of Mongolian head and face in China
    LI Yong-lan LIU Lu
    2019, 50 (1):  98-106.  doi: 10.16098/j.issn.0529-1356.2019.01.017
    Abstract ( )   PDF (1248KB) ( )  

    Objective To explore the physical anthropology characteristics of Chinese Mongolian head and face. Methods A total of 12 head and face indexes (head length, head breadth, face breadth, minimum frontal breadth,morphological facial height, nasal height, nasal breadth, interocular breadth, physiognomic ear length, lip height, mouth breadth, auricular height) of the 17 Mongolian ethnic groups were analyzed by the cluster analysis and principal component analysis. Results The head length, face breadth and nose breadth of Duerbote, Guoer Ross, Fuxin, Calaqinleft were small, and the interocular breadth was close. Northern Han was close to Horqin. The head breadth, face breadth, nose breadth of Bargud and Ordos were large but mouth breadth, minimum frontal breadth, interocular breadth were small. Buriat, Torghut, Chifeng, Northeast Han and Xilincol had something in common that head, face and nose were more broad, and the mouth breadth, minimum frontal breadth and interocular breadth were large. Head length, face breadth, nose breadth of Xinjiang Qahar and Torghut were small, but mouth breadth, interocular breadth and minimum frontal breadth were medium. The Mongolian ethnic group’s facial features in Yunnan province differed greatly from the Mongolian ethnic groups in the north. Conclusion According to the characteristics of head and face, Mongolian people can be divided into three groups. The principal component analysis shows that the width index of head and face is the main basis for distinguishing the facial features of all ethnic groups in Mongolian. The facial features of Northern Han and Mongolian have some commonalities. Generally speaking, Mongolian ethnicity morphological facial height and, nasal height are close to East Asia but not North Asia.

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    Vital capacity and body composition of multi national college students in Guangxi
    GONG Jian-gu HUANG Li-qian TANG Peng QIN Hua-xiao DENG Tuo HUANG Fang YUAN Chao-he ZHENG Qing-song XU Lin LIU Peng
    2019, 50 (1):  107-110.  doi: 10.16098/j.issn.0529-1356.2019.01.018
    Abstract ( )   PDF (1190KB) ( )  

    Objective To study the basic situation of vital capacity and body composition of the multi ethnicity college students in Guangxi, and to explore the correlation between vital capacity and body composition. Methods The vital capacity and body composition of 1322 students who volunteered to participate in the investigation in a college in Guangxi were measured, and carried on the statistical analysis. Results There were ethnic differences in vital capacity, muscle volume, estimated bonse mass, body moisture and protein among the Han, Zhuang and other ethnic minority students (P<0.05). In the correlation analysis, the vital capacity and the amount of fat were negatively correlated (correlation coefficient r=-0.092, P<0.01). The vital capacity and muscle mass, the presumption of bone mass, body moisture and protein content were positively correlated (correlation coefficients were 0.687,0.633,0.658,0.653,respectively and P<0.01).Conclusion Ethnic differences are in vital capacity and some body composition among ethnic university students in Guangxi, and the comparison of the vital capacity and body composition among the three are: Han>Other minorities>Zhuang. The higher lung capacity, the lower the amount of fat in the body, and the higher muscle mass, bone mass, body moisture and protein content.

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     Soft tissue facial profile of Han nationality young students for different somatotypes in Zibo Shandong
    SUN Zhao-xia CHENG Hui WU De-ye
    2019, 50 (1):  111-118.  doi: 10.16098/j.issn.0529-1356.2019.01.019
    Abstract ( )   PDF (1246KB) ( )  

    Objective To observe the differences of somatotype, facial tissue profile between male and female of Han students, and the differences of facial tissue profile in homogeneous somatotype between male and female. Methods A total of 326 males and 478 females who were 18-23 years old with no disability, disease, deformity and no facial plastic history were studied. Somatotype was assessed by Heath-Carter method , and the angles of facial tissue profile were measured by FreeHand MX software. Results Han young males had central somatotype (3.52-3.28-3.40), and females with balanced endomorphy (4.52-3.03-3.23) that facial profile was to accord with esthetic Ricketts and Steiner plane. For total samples, it was negative correlation between height, body weight, tensive biceps girth, calf girth, humerus width and frontonasal angle, it was positive correlation between height and nasal tip angle, tensive biceps girth and nasolabial angle.It was negative correlation between height and mentolabial angle, between body weight, calf girth, endomorphic factor and fontonasal angle. It was positive correlation between ectomorphic factor and fontonasal angle for males homogeneous somatotype (P<0.05). It was negative correlation betweenendomorphic factor and nasolabial angle.It was positive correlation between endomorphic factor and nasal tip angle for females homogeneous somatotype (P<0.05). It was negative correlation between years of age and nasolabial angle for males and females, and mentolabial angle for females(P<0.05). Conclusion Frontonasal angle is different between Han young students. It is different that somatotype influence soft tissue facial profile between Han young students. Frontonasal angle, nasal tip angle, lip convex angle are relative stable with years of age to change for Han young students.

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    Analysis and evaluation of Heath-Carter somatotype based on dorsal muscle strength
    LIU Yuan-yuan YIN Shuai SUI Yue-lin SUN Yang PANG Yin LU Lan-hong MU Hong-mei
    2019, 50 (1):  119-122.  doi: 10.16098/j.issn.0529-1356.2019.01.020
    Abstract ( )   PDF (793KB) ( )  

    Objective To explore the somatotype classification of high level dorsal muscle strength. Methods Dorsal muscle strength of a complete training unit of 418 male soldiers, according to the provisions of GJB1337-92 were measured and classified the somatotype by the method of Heath-Carter. Results The mean value of dorsal muscle strength of the 418 male soldiers was (134.3±22.6) kg, overall evaluation was good. Dorsal muscle strength good and above was 270, accounting for 64.7%; the mean value was (147.0±16.7) kg, the mean value of somatotype was 2.4(0.8)-5.0(1.2)-2.3(0.9), somatotype attitudinal mean(SAM)was 1.5, 87.8% focused on the medium of embryo type. The factors of endomorphy, mesomorphy and ectomorphy were the main factors that affected the dorsal muscle strength score. The better of the dorsal muscle strength score, the lower value of the endomorphy and ectomorphy factors, the higher value of the mesomorphy factor. Conclusion The high level of dorsal muscle strength in the somatotype classification is highly concentrated in the medium of the embryo type. Dorsal muscle strength is significantly positively correlated with the mesomorphy value.

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    Review
    Role of histone variants in the reprogramming of parental genomes
    WANG Nan HUANG Xing-wei CHENG Xiang-rong JIANG Qi PANG Nan LEI Lei
    2019, 50 (1):  132-136.  doi: 10.16098/j.issn.0529-1356.2019.01.023
    Abstract ( )   PDF (1172KB) ( )  

    Oocytes have the ability to reprogram the sperm genome to ensure normal embryonic development. After sperm enters into the egg, the paternal genome undergoes protamine to histone exchange, whole genome demethylation and other processes, which start embryonic development. Histone H3 variant H3.3 can replace canonical histones H3.1 and H3.2 in nucleosomes, thereby modifying chromatin structure and regulate gene expression. The absence of H3.3 during early embryonic development will lead to excessive chromosomal aggregation and mis-isolation. This article reviews the role of histone variant H3.3 and its chaperones in spermatogenesis, fertilization and early embryonic development. In particular, the importance of H3.3 in the reprogramming of the paternal genome is of great significance in understanding the formation of totipotent zygotes after fertilization and their preimplantation development.

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    Neurobiology
    Apoptosis of olfactory bulb cells in MPTP cynomolgus monkey model of Parkinson’s dise 
    CHEN Ming-yu CHENG Wei YAN Li-li GUAN Yun-qian WU Di YUE Feng REN Zhen-hua
    2019, 50 (1):  3-7.  doi: 10.16098/j.issn.0529-1356.2019.01.001
    Abstract ( )   PDF (3333KB) ( )  

    Objective To explore the apoptosis and the proliferation of glia cells in olfactory bulb of cynomolgus monkeys damaged by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Methods Three adult cynomolgus monkeys were injected with MPTP to induce the damage of olfactory bulb as MPTP group (model group) and other three adult cynomolgus monkeys as saline group (control group). Immunohistochemical staining was performed to examine the expression of Caspase-3, Bcl-2, ionized calcium binding adaptor molecule-1(Iba-1) and glial fibrillary acidic protein(GFAP) in the olfactory bulb. The numbers of Caspase-3, Bcl-2, Iba1 and GFAP-positive cells were counted and the average absorbance was measured by Image J v1.8.0 in control and MPTP group. Results Compared with the normal control group, Caspase-3-positive cells increased significantly in the glomerular layer (GL) of olfactory bulb, and Bcl-2-positive cells decreased significantly in MPTP group after MPTP injury, and GFAP-positive and Iba-1-positive cells were located in GL and external plexiform layers (EPL). The number of GFAP-positive and Iba-1-positive cells increased in olfactory bulb in the MPTP group. Conclusion MPTP can induce the cell apoptosis, astrocyte and microglial activation in olfactory bulb of cynomolgus monkeys, which may be associated with the dysosmia in Parkinson’s disease.

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    Analysis of the difference in gene expression and mutation of glioblastoma in different age groups by using the transcriptional and exome sequencing
    WANG Zhi-gang LIU Wei
    2019, 50 (1):  8-12.  doi: 10.16098/j.issn.0529-1356.2019.01.002
    Abstract ( )   PDF (6853KB) ( )  

    Objective To explore the gene expression difference of glioblastoma in different age groups through the sequencing of the transcriptome and exome. Methods Glioblastoma multiforme(GBM) data was downloaded from the Cancer Genome Atlas(TCGA), and the samples were divided into two groups. There were 78 cases in the middle and low age group not more than 60 years old, compared with the high age group of 60 years old, with a total of 76 cases. DESeq2 package was used to analyze differentially expressed genes based on gene level reads numbers. Cytoscape plug-in ClueGO was used to analyze gene function enrichment in different age groups. Results Most of the gene expression trends in the two groups were consistent between the two groups, but there were some different genes. In the middle and low age group, the differential gene was mainly related to the proliferation of neural precursor cells, while the high age group was metabolic. In the two groups, the high mutation genes were basically consistent, but the mutation rates were different. The mutation differentially genes were concentrated in the middle and low age groups. Conclusion There is a significant difference in gene expression and gene mutation between the middle and low age group and the elderly GBM group.

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    Dynamic observation of injury of HT22 cell after oxygen and glucose deprivation/reoxygenation
    ZHANG Yi ZHOU Xiao-hong JIN Xiao-fei ZHANG Ying GAO Wei-juan
    2019, 50 (1):  13-17.  doi: 10.16098/j.issn.0529-1356.2019.01.003
    Abstract ( )   PDF (5501KB) ( )  

    Objective To investigate the damage of hippocampal neuron cell line HT22 at different time points after oxygen and glucose deprivation/reoxygenation. Methods Logarithmic growth phase of HT22 cells was randomly divided into six groups, normal control group,OGD/R 0 hour, OGD/R 12 hours, OGD/R 24 hours, OGD/R 48 hours,OGD/R 72 hours groups. All groups were treated with reoxygenation after deprived of oxygen and glucose for 6 hours except normal group.The cell morphology was observed by invertded microscope. Cell counting kit-8(CCK-8) method was used to test the cell survival rate. Lactate dehydrogenase (LDH) method was used to detect cell damage. Bax and Bcl-2 immunofluorescence were used to detect apoptosis. Results HT22 cells in normal group were bipolar or multipolar. Obvious synapses and multiple synapses were woven into a network. Cell membrane was smooth and complete and the refractivity of cell was strong. But the axons decreased, cell was shrinkage and cytoplasmic aggregation in HT22 in OGD/R groups, and the cell viability was significantly reduced(P<0.05), the lowest point appeared in OGD/R 12 hours to OGD/R 24 hours; OGD/R 12 hours and OGD/R 24 hours LDH increased significantly (P<0.05), OGD/R 0 hour, 48 hours, and 72 hours increased. The ratios of Bax/Bcl-2 in OGD/R groups were significantly increased (P<0.05) except for OGD/R 0 hour and OGD/R 72 hours, and peaked at OGD/R 24 hours. Conclusion Injury of cells afer OGD/R is a complex and dynamic process. Cell injury is aggravating as the reoxygenation time increased and reached a peak in OGD/R 24 hours, from then on, cell damage gradually decreased.

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    Cell and Molecules Biology
    Gastrodin inhibits apoptosis of H9c2 cardiomyocytes through protein kinase B/p38mitogen actived protein kinase signaling pathway
    ZHANG Ling YANG Ping JIANG Yong-liang PU Yu-wei XIE Li-qiu SUN Lin LU Di
    2019, 50 (1):  40-48.  doi: 10.16098/j.issn.0529-1356.2019.01.008
    Abstract ( )   PDF (7764KB) ( )  

    Objective To investigate the effect and precise molecular mechanisms for explaining how gastrodin suppresses the apoptosis in the serum deprivation/reperfusion-induced H9c2 cardiomyocytes. Methods Rat H9c2 cardiomyocytes were cultured and treated differently in vitroand randomly divided into following groups, including the control group, the serum deprivation-stimulated (0.5-6 hours) groups, the serum deprivation/reperfusion(2/4 hours) group, gastrodin (10, 20 and 40 μmol/L) treatment groups, wortmannin (1 μmol/L) and gastrodin(20 μmol/L) co-treatment group, wortmannin (1 μmol/L) treatment group. The variation of protein expression levels of p-p38 mitogen actived protein kinase(MAPK), p-Akt and apoptosis related cleaved caspase-3, Bcl-2 and Bax were analyzed by Western blotting.H9c2 cardiomyocytes were analyzed by flow cytometry.The level of RNA expression levels of apoptosis related Caspase-3,Bcl-2 and Bax were analyzed by Real-time PCR assay. The expression level of p-Akt was assayed by double immunofluorescence labeling. Results The results showed that serum deprivation (0.5-6 hours) and serum deprivation/reperfusion (2/4 hours) induced apoptosis increasing; Using Western blotting and Real-time PCR assay, we found that treatment with gastrodin significantly alleviated cleaved Caspase-3 activation and increased the ratio of Bcl-2/Bax in a concentration-dependent manner after serum deprivation and serum deprivation/reperfusion exposure. The same result has been showed by flow cytometer. Serum deprivation and serum deprivation/reperfusion inhibited the Akt/p38MAPK signaling pathway; After treatment with gastrodin in a concentration-dependent manner induced the expression of p-Akt and decreased the expression of p-p38MAPK. The regulation effects of gastrodin were reversed by the Akt specific inhibitor wortmannin. Conclusion In conclusion , the present study demonstrated that gastrodin exerted protective effects against serum deprivation/reperfusion-induced apoptosis in H9c2 cardiomyocytes, at least partly via the Akt/p38MAPK pathway.

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    Protective effect of flavonoids of rosa roxburghii tratt on myocardial cytotoxicity induced by doxorubicin
    YUAN Hui-fang ZHANG Yong-chun CAI Xin-hua XU Ping CHEN Hui
    2019, 50 (1):  49-55.  doi: 10.16098/j.issn.0529-1356.2019.01.009
    Abstract ( )   PDF (16160KB) ( )  

    Objective To investigate the effect and mechanism of flavonoids of rosa roxburghii tratt(FRRT) on apoptosis in doxorubicin(DOX)-induced cardiotoxicity. Methods Myocardial cells were isolated and cultured from 26 newborn rats of 1-3 days or H9C2 myocardial cell line. The myocardial cytotoxicity model was established by incubating myocardial cells with a concentration of 5 μmol/L DOX, and FRRT group were intervened with FRRT incubation in advance. Ultrastructure of H9C2 myocardial cells were observed with transmission electron microscope. The expressions of Bax, P53 and Bcl-2 in myocardial cells were detected with Western blotting and immunofluorescence, and the results of Western blotting were analyzed with semi-quantitatively. Results Transmission electron microscopy showed that myocardial cells had a large number of autophagic vacuoles and lipofuscin, and cells were in poor condition in DOX group, the myocardial cells of the FRRT+DOX group had no structure of lipofuscin, and only a small amount of autophagic vacuoles. The results of immunofluorescence showed that the expressions of Bax and P53 were positive, and Bcl-2 was weakly positive in DOX group. The expression of Bax and P53 were not obvious, and Bcl-2 was strongly positive in FRRT+DOX group. The results of Western blotting showed that the myocardial cells were incubated with DOX for 18hours, the expression level of Bax was the highest, and the difference had statistical significance between groups (P<0.01).The expressions of Bax were increased in DOX group,and reduced in FRRT+DOX group, and the difference had statistical significance between groups (P<0.05). The expressions of P53 were increased in DOX group,and reduced in FRRT group, and the difference without statistial significance between groups(P>0.05). The expressions of Bcl-2 were reduced in DOX group, and increased in FRRT+DOX group, the difference had statistical significance between groups(P<0.05, P<0.01). Conclusion FRRT protects cardiomyocytes by down-regulating DOX-induced cardiomyocyte apoptosis.

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    Neurobiology
    Fluoxetine improving the depression-like behavior induced by chronic restraint stress by up-regulation the expression of bromodomain-containing protein 4 in hippocampus of mouse
    WANG Zhen HUANG Yi-jia NAI Ai-tao NIU Lei LUO Shi-shi WAN Wei LIU Zheng-hai CHEN Xi XU Yang CAO Wen-yu
    2019, 50 (1):  18-23.  doi: 10.16098/j.issn.0529-1356.2019.01.004
    Abstract ( )   PDF (1320KB) ( )  

    Objective To investigate the effect of fluoxetine (FLX) on the expression of bromodomain-containing protein 4 (BRD4) in the hippocampus induced by chronic restraint stress (CRS), with the depression-like behaviour also being determined. Methods Twenty-four male Kunming mice were randomly divided into normal saline(NS) group, CRS group and fluoxetine(FLX) intervention(CRS+FLX) group. The mice of the CRS group were subjected to 3 weeks chronic restraint stress. The mice of CRS+FLX group were treated with fluoxetine by intraperitoneal injection 30 minutes before restraint stress from the 8 to 21 days. The mice in the NS group and CRS group were treated with NS. Depression-like behavior was determined by sucrose preference test, sucrose splash test, forced swimming test, novelty object recognition test and open field test. Expression of BRD4 in the hippocampus was determined by Western blotting and Real-time PCR. Results Compared with NS group, mice in the CRS group showed reduced sucrose preference (P<0.01) and decrease the time spent licking in Sucrose splash test (P<0.05) , increase immobility time in the forced swimming test (P<0.01) and decrease on the discrimination ratio in the novelty object recognition test, while FLX treatment effectively reversed the depression-like behavior induced by CRS (P<0.05) Compared with the NS group, CRS led to significantly decreased expression of BRD4 protein and mRNA in the hippocampus (1.000 ± 0.04577 vs 0.08337 ± 0.01658; 1.000 ± 0.04379 vs 0.6672 ± 0.03193, P<0.05), which was attenuated by FLX treatment (0.08337 ± 0.01658 vs 0.4983 ± 0.08574; 0.6672 ± 0.03193 vs 0.8572 ± 0.03181, P<0.05). Conclusion Our finding indicates that FLX could alleviate CRS-induced depression-like behavior in mice, which might be attributed to the increased expression of BRD4 in the hippocampus.

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    Optimization of establishment of pig pluripotent cells and directly differentiation into neural lineage cells
    LI Xue ZHANG Ben NIU Shu-dong WANG Yu-ge WEN Li-bo LIANG Chen QI Xiao-juan LI Yu LEI Lei
    2019, 50 (1):  24-28.  doi: 10.16098/j.issn.0529-1356.2019.01.005
    Abstract ( )   PDF (3370KB) ( )  

    Objective To generate the induced pig pluripotent cells (iPPCs) using the optimized induced pluripotent stem cell(iPSc) technology, and to discuss the method of directly differentiate into neural lineage cells. Methods we generated the iPPCs using the classical iPS technology in combination with valproic acid (VPA) and the methyltransferase inhibitor 5-aza-2 -deoxycytidine (5-AZA) with Oct4 retrovirus infected repetitively. Real-time PCR analysis of the expression of pluripotent related genes, immunofluorescent detection of neuron specific marker after using retinoic acid (RA) and extracellular matrix induction. Results By using this optimizing iPS technology, we successfully established a reasonable method to generate iPPCs. Real-time PCR result indicated VPA treatment significantly increased the expression of pluripotent genes, which was the same as repeated infection with Oct4 retrovirus. In addition, the iPPCs might directly differentiate into neural lineage cells after being induced with the retinoic acid and extracellular matrix. Conclusion We establishes a reasonable method to generate pig pluripotent cells, which may be a new donor cell source for human neural disease therapy.

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    Neurobiology
    Granulocyte colony stimulating factor improving the learning and memory function and its anti-inflammatory mechanism of Alzheimer’s disease model rats
    SU Xiang ZHANG Zhen YANG Lin-jing QIU Yi-hui SONG Cheng-long CHEN Juan-yan SONG Shui-jiang
    2019, 50 (1):  29-34.  doi: 10.16098/j.issn.0529-1356.2019.01.006
    Abstract ( )   PDF (2999KB) ( )  

    Objective To investigate the effect of granulocyte colony-stimulating factor (G-CSF) on cognitive impairment in Alzheimer’s disease(AD) in rats and its effect on brain inflammatory response. Methods Forty-five male Sprague-Dawley rats were randomly divided into 3 groups: sham operation group, model group, and G-CSF group. Rat models were induced by intracerebro ventricular injection of amyloid beta-peptides 1-42 (Aβ1-42), and then G-CSF were given to G-CSF group. The learning and memory function and brain histopathological changes were observed and compared. The expression of nuclear factor κB p65 (NF-κB p65), phosphorylated p38 mitogen activated protein kinase (p-p38MAPK), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor α(TNF-α) and interleukin-1β(IL-1β) were detected by Western blotting. The levels of iNOS, COX-2, TNF-α and IL-1β mRNA were detected by Real-time PCR. Results The number of neurons in the model group decreased significantly, the nuclear condensation was obvious and the nucleolus was unclear. Neuronal nuclear condensation was significantly improved in G-CSF group. The escape latency of the sham group, model group and G-CSF group were (35.68±6.73) s, (57.92±7.35) s and (40.27±8.91)s. The swimming time of the target quadrant was 54.72%±4.22%, 36.73%±3.2% and 44.68%±4.01%. The times of crossing the platform were 8.7±2.1, 3.9±1.6 and 6.5±1.7. Compared with the model group and the control group, the escape latency of G-CSF group were significantly shorter, target quadrant swimming time increased significantly, through the platform number increased significantly, the difference had statistical significance (P<0.05).In the sham group, model group and G-CSF group, the protein levels of NF-κB p65 were 0.144±0.033, 0.502±0.035 and 0.473±0.061, the protein levels of p-p38MAPK were 0.194±0.021, 0.511±0.039 and 0.266±0.048. Compared with the model group, the levels of F-κB p65 and p-p38MAPK protein in the sham group and G-CSF group decreased significantly, and the difference was statistically significant(P<0.05). Compared with the model group, the protein and mRNA levels of iNOS, COX-2, TNF-α and IL-1β in the sham group and G-CSF group were significantly lower than those in the model group (P<0.05).Conclusion G-CSF can improve the learning and memory dysfunction of rats with Alzheimer’s disease probably through inhibiting the activation of p38 MAPK and NF-κB p65 signaling pathways and down-regulating iNOS, COX-2, TNF-α and IL-1β expressions to inhibit the neuroinflammatory state of the brain.

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     Inhibition of the aquaporin4 correlates with the analgesic effects via extracellular regulated protein kinase pathway activation of spinal dorsal horn after sciatic nerve injury
    ZHAO Liang LI Dan LIU Nan LIU Lu LI Hong-peng
    2019, 50 (1):  35-39.  doi: 10.16098/j.issn.0529-1356.2019.01.007
    Abstract ( )   PDF (4558KB) ( )  

    Objective To investigate the effect of inhibiting aquaporin 4(AQP4) on the activation of glial cells and mitogen-activated protein kinase(MAPK) signaling pathway in sciatica caused by lumbar disc herniation. Methods Male SD rats(n=90) were divided into sham group(n=18), chronic compressive injury(CCI)+DMSO group(n=36) and CCI+TGN-020 group(n=36). Make CCI model. Test sample of spinal by Western blotting method and immunology chemical fluorescence double staining method . Results Western blotting showed the astrocytes and extracellular regulated proptein kinase(ERK), c-Jun N-terminal kinase(JNK), p38MAPK pathway were activated. Immunofluorescence found TGN-020 could inhibit the expression of AQP4, weaken the activation of glial cells induced by nerve injury, and affected the activation of ERK,JNK and p38MAPK signaling pathways. We also found that the number of cells expressing p-ERK and glial fibrillary acidic protein(GFAP) at the same time increased obviously after injury, and could be significantly reduced by TGN-020. Conclusion Inhibition of AQP4 can inhibit the activation of astrocyte and activation of MAPK signaling pathway of spinal dorsal horn, which are caused by sciatic nerve injury. Inhibition of AQP4 can reduce the activation of astrocyte by inhibiting activation of the ERK pathway in the astrocytes in spinal dorsal horn, and improving the neuropathic pain caused by sciatic nerve injury.

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    Histology,Embryology and Developmental Biology
    Imatinib mesylate regulating the effect of Telocytes on salivary secretion of submandibular gland
    PENG Li-juan HUANG Yue YAN Hui-wen GAO Jie HU Rong SU Min
    2019, 50 (1):  91-97.  doi: 10.16098/j.issn.0529-1356.2019.01.016
    Abstract ( )   PDF (14887KB) ( )  

    Objective To explore the effect of Telocytes (TCs) on salivary secretion of submandibular gland by imatinib mesylate’s regulation. Methods The TCs immune-labellings C-kit/CD117 and CD34 were double labelled with immunofluorescent staining technology to localize TCs in the submandibular gland of mice. The transmission electron microscopy (TEM) method was used in the present study to observe the ultrastructure of TCs and their relationship with neighboring cells in the submandibular gland of mice. The mice were divided into 5 groups:normal group(24), 1-week, 2-week, 3-week and 4-week medicine groups(24),which were administered medicine at 80 mg/(kg·d) daily in order to study the effect of imatinib mesylate treatment on TCs function. Immunofluorescent staining technology was used to observe the change of TCs. Western blotting was used to evaluated the change of level of CD117/CD34/α-amylase(α-Amy)protein expression. Results The immunofluorescent staining showed that TCs were widely distributed in connective tissues in submandibular gland and cellular body was small with protrusions, or telopodes(Tps). Tps were connected each other and formed network-like structure, surrounding acinus and ducts. The network structure consisting of TCs protrusions in the medication groups were gradually reduced with the time of experiment. The electron microscopy observation showed that the TPs possess like-moniliform structure, connecting with adjacent tissues and being surrounded by extracellular vesicles. The number of TPs and the intracellular organelles decreased but the intracellular vesicles increased with the time of experiment. Western blotting showed that the level of CD117/CD34/α-Amy protein expression were also declined, and positively correlated with each other. Conclusion There are TCs in the submandibular gland of mice. Intervention of TCs with imatinib mesylate may reduce the salivary function of submandibular gland by affecting the structure, immunolabel and intercellular communication of TCs.

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    Anatomy
     Intramuscular nerve distribution pattern of anterolateral abdominal wall muscles
    YANG Xian ZHAO Hong-ming YANG Sheng-bo
    2019, 50 (1):  77-81.  doi: 10.16098/j.issn.0529-1356.2019.01.013
    Abstract ( )   PDF (6970KB) ( )  

    Objective To investigate the intramuscular nerve distribution pattern of anterolateral abdominal wall muscles and to explore its clinical significance. Methods Twenty-four anterolateral abdominal wall muscles from twelve Chinese adult cadavers that were fixed with formaldehyde were removed for modified Sihler’s staining. Results Most of the innervation of the anterolateral abdominal wall muscles originated from the segmental distribution of the intercostal nerve. Each muscular serration of the obliquus externus abdominis muscle had an independent innervation and there was a single longitudinal nerve dense band above the iliac crest between the anterior superior iliac spine and iliac tubercle. In the obliquus internus abdominis muscle, there was a longitudinal nerve dense band above the midpoint of the inguinal ligament and a transverse nerve dense band above the iliac crest. In the transversus abdominis muscle, there was an arched nerve dense band between the anterior superior iliac spine and the junction of the ninth and eighth costal cartilage. The rectus abdominis muscle had one transverse nerve dense band in the middle of each muscle belly and the upper 3 muscle bellies had independent innervation. Conclusion The obliquus externus abdominis and rectus abdominis muscle can be divided into neuromuscular subdivisions. The nerve dense zone in each muscle should be considered as the optimal target site for botulinum toxin A injections and local anesthesia, and it should not be designed as the incision site for abdominal wall surgery.

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    Anatomic morphology measurement and clinical significance of scapular spine based on CT 3D reconstruction
    ZHANG Lei LIU Yang ZENG Yan LI Jing YI Gang GUO Xiao-guang ZHANG Yong-tao FU Shi-jie
    2019, 50 (1):  82-86.  doi: 10.16098/j.issn.0529-1356.2019.01.014
    Abstract ( )   PDF (2134KB) ( )  

    Objective To research anatomic morphology and clinical significance of scapular spine(SS) based on CT three-dimensation reconstruction. Methods A total of 180 SS (left 90, right 90) data collected from Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University were measured based on CT three-dimensation reconstruction, including the morphological characteristics, the shape of the SS, SS length, the width of the central and base length of SS, the width and height of eminence of SS. Results The SS was superficial and constant. The average length of SS and basement were (128.31±8.59)mm and (80.35±6.70)mm respectively. The central width of SS was(9.69±2.48)mm, the average width and height of eminence of SS were (8.98±2.68)mm and (13.48±2.92)mm, there was no statistical difference between sides of body (P>0.05). Conclusion CT three-dimensation reconstruction has unique advantages in anatomical morphology research of SS, offering substantial benefits for the orthopedist in preoperative planning, and using precontoured locking plates may be an additional aid during surgery, so as to reduce intraoperative risks.

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    Origin and significance of coronary artery
    ZHANG Tie-shan LOU Li-fang ZHENG Lin-lin MA Zhong-liang XU Ying WANG Zheng XU Gao-lei ZHANG Zhen-hua
    2019, 50 (1):  87-90.  doi: 10.16098/j.issn.0529-1356.2019.01.015
    Abstract ( )   PDF (4541KB) ( )  

    Objective To describe the normal and variant anatomy of the coronary artery ostia in Chinese subjects, and to provide data reference for coronary angiography and intervention. Methods Fifty-five heart specimens were dissected. The number of ostia and their positions within the respective sinuses were observed. Vertical and circumferential deviations of the ostia were observed. The heights of the cusps, the ostia and sinutubular ridge from the bottom of aortic sinus were measured and recorded, analyzed and reported the result . Results The number of openings in the aortic sinuses varied from 2-5 in the present series; multiple ostia were mostly seen in the anterior sinus. The majority of the ostia was below the sinutubular ridge (94.5%) and at or above the level of the cusps (80%). No openings were present in the pulmonary sinuses and posterior aortic sinus. There were high openings in 5 specimens, which were higher than the sinutubular junction. Conclusion Most of the coronary openings are located in the right aortic sinus or left aortic sinus, above the cusps and below sinutubular ridge. Some mutations, such as multiple openings, longitudinal or circumferential positional shifts, and slit-like openings can cause difficulties in interpreting images, and adversely affect clinical operations such as angiography, angioplasty and coronary artery bypass grafting.

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    A rat model of intrauterine adhesion established by endometrial scraping
    CHEN Xing MAO Le-le DIAO He SUN Yu QIN Li-hua BAI Wen-pei
    2019, 50 (1):  123-127.  doi: 10.16098/j.issn.0529-1356.2019.01.021
    Abstract ( )   PDF (13251KB) ( )  

    Objective Intrauterine adhesion (IUA) seriously affects reproductive health of women. In order to better study IUA, we established a rat model of IUA by endometrial scraping, and evaluated the method of this model. Methods A total of 60 SD rats were randomly divided into three groups: the sham group, the endometrial layer scraped group and the endometrial and partly mesenchymal layer scraped group (n=20 each). In the sham group, uteri were cut open and then closed without any other surgical procedures. In the other two groups, endometrial and mesenchymal layers of rats were scraped and sutured. After 3, 7, 14, 21 and 28 days of scraping, uteri were sampled for histological and immunochemical evaluation. Results The endometrial layer of uteri was disappeared. The new endometrial layer of uteri was seen in the endometrial layer scraped group after 7 days of scraping. In the endometrial and partly mesenchymal layer scraped group, folds of endometrium were absent, the endometrial layer was removed, at least 50% of mesenchymal layer was disappeared, the number of endometrial glands were significantly decreased, and the fibrosis area of endometrium was increased with the development of fibrosis. The expression of transforming growth factor β1 was increased while matrix metalloproteinase 9 was decreased, which was in accordance with pathophysiological changes.
    Conclusion Endometrial and partly mesenchymal layer scraping can establish an IUA rat model. The IUA rat model is in agreement with IUA changes, which can be used in IUA studies.

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    Technology and Methodology
    Establishment of a diabetic lower extremity arterial disease rat model of restenosis after percutaneous transluminal angioplasty
    ZOU Zhi-wei ZHANG Qian DONG Jian-jun
    2019, 50 (1):  128-131.  doi: 10.16098/j.issn.0529-1356.2019.01.022
    Abstract ( )   PDF (6100KB) ( )  

    Objective To establish an animal model of restenosis in diabetic lower extremity arterial disease rat after percutaneous transluminal angioplasty(PTA). Methods We divided diabetic rats(n=30) into three groups: atherosclerosis group(AS), restenosis group(RS) and control group. We examined the artery by hematoxylin and eosin(HE), and detected the plaque components by αsmooth muscle actin(α-SMA) immunohistochemical staining. Results Artery restenosis rat models were successfully developed. The smooth muscle cells (SMCs) were the key components of the RS plaque. Conclusion An experimental animal model of restenosis in diabetic lower extremity arterial disease rat after PTA is set up by the steps described in this study.

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