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    Cell and Molecules Biology
    Endothelial differentiation potential of neonatal rat cardiac fibroblasts
    ZHANG Jun-yue YIN Guo-tian GUO Zhi-kun
    2018, 49 (4):  443-449.  doi: 10.16098/j.issn.0529-1356.2018.04.005
    Abstract ( )   PDF (1183KB) ( )  

    Objective  To investigate the differentiation of cardiac fibroblasts (CFSs) into endothelial cells and the potential for vascularization, and to provide a cytological and theoretical basis for clinical treatment of myocardial injury. Methods  CFSs were isolated, cultured and purified from the fresh left ventricular tissue. The 3rd generation of CFSs was induced by endothelial cells induced medium for 28 days. After continuous induction culture of 28 days, the endothelial cell medium (ECM) was replaced, digested and passaged down to P3 generation. The growth and morphological changes of the induced cells were detected. The expression of the endothelial cell markers and functional characteristics in the induced cells were evaluated by immunocytochemistry, flow cytometry and angiogenesis analysis. Results  The third generation of CFSs in neonatal rats was triangular, spindle and polygonal, and the rate of proliferation was rapid. Vimentin and discoidin domain receptor 2(DDR2) expressions were positive. The 3rd generation of CFSs was cultured for 3 hours and the induced medium was added, cells began to converge after 3 days, the cells formed string beads were induced at 21 days, and the cells were pooled into the ring shape at 28 days. The immunohistochemistry staining showed that vWF and CD31 were positively expressed in the differentiated cells, but not expressed in the control group. The immunofluorescence staining shows that vWF, CD34 and CD105 were positive in the experimental group, but not in the control group. Flow cytometry analysis showed that the expression rate of CD31 was 50.5% in the induced cells, whereas the expression in the control group was just 5.82%. Conclusion  Cardiac fibroblasts induced with vascular endothelial growth factor(VEGF) and basic fibroblast growth factor(bFGF ) can differentiate into vascular endothelial cells in vitro, and have the characteristics and functions of vascular endothelial cells.

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    Cancer Biology
    TRIM28siRNA enhancing the sensitivity of non-small cell lung cancer cell PAa to etoposide through activating E2F transcription factor 1
    LIU Lei GAO Ya-xian WANG Jian-ping ZHAO Xue-rong XU Qian HU Wang LIU Mei-qi
    2018, 49 (4):  455-460.  doi: 10.16098/j.issn.0529-1356.2018.04.007
    Abstract ( )   PDF (400KB) ( )  

    Objective  To investigate the effect of small interfering RNA (siRNA)-mediated silencing of TRIM28(tripartite motif containing 28)on the sensitivity of non-small cell lung cancer cell (NSCLC) PAa to etoposide and explore the possible mechanism. Methods  The expression of TRIM28 in PAa cell was silenced by RNA interference. MTT and colony formation assay were used to assess the inhibitory effect of TRIM28siRNA alone or in combination with etoposide on the growth and proliferation of PAa cells. The apoptosis of different groups was detected by flow cytometry(FCM ) and the expression of E2F transcription factor 1(E2F1) was evaluated by Western blotting. Results  A PAa cell line silenced with the TRIM28siRNA was established. TRIM28 in combination with etoposide significantly inhibited the proliferation and colony formation of PAa cells. TRIM28siRNA in combination with etoposide induced apoptosis and increased the expression of E2F1 at mRNA and protein levels. Conclusion  TRIM28siRNA in combination with etoposide has an obvious effect on the growth of NSCLC cells, suggesting a new clinical treatment strategy for NSCLC.

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    MicroRNA-539 inhibiting the occurrence and development of hepatocellular carcinoma by targeting E2F transcription factor 3
    DENG Xue-jun GONG Shao-xin YANG Sheng-hui HUANG Li-rong ZHOU Fang
    2018, 49 (4):  469-474.  doi: 10.16098/j.issn.0529-1356.2018.04.009
    Abstract ( )   PDF (307KB) ( )  

    Objective To investigate the expression of microRNA-539 (miR-539) in hepatocellular carcinoma (HCC) and the possible mechanism of miR-539 inhibition cancer cells. Methods  Ninety cases of liver cancer patients after liver cancer tissue and adjacent tissue specimens were collected, and the expression of miR-539 in clinical tissues was detected by Real-time PCR. MiR-539 mimics was transfected into QGY-7701, Huh7 and PCL/PRF5 hepatoma cells, and the expression of miR-539 was detected by Real-time PCR. The effect of miR-539 on the proliferation of HCC cells was detected by MTT assay. The effect of miR-539 on the apoptosis of hepatocellular carcinoma cells was detected by flow cytometry. The effect of miR-539 on the expression of E2F transcription factor 3(E2F3) protein in hepatocarcinoma cells was detected by Western blotting. Results  The expression of miR-539 in hepatocellular carcinoma was significantly lower than that in adjacent tissues. The expression of miR-539 was not correlated with sex, age and histological differentiation of hepatocellular carcinoma (P>0.05). The expression of miR-539 was correlated with tumor diameter and lymph node metastasis (P<0.05). The expression of miR-539 in miR-539 mimics group was significantly higher than that in blank group and miR-539 NC group (P<0.05). In QGY-7701 cells, overexpression of miR-539 significantly inhibited cell proliferation (P<0.05), decreased E2F3 protein expression (P<0.05), but was no significant effect on apoptosis (P>0.05). Conclusion  miR-539 is a tumor suppressor factor in the development of hepatocellular carcinoma, which may inhibit the proliferation of cancer cells through the down-regulation of E2F3 protein level.

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    Cell and Molecules Biology
    MiR-382 promoting BRL-3A cell proliferation of rat
    GAO Hang ZHANG Chun-yan WHANG Feng-juan XU Cun-shuan
    2018, 49 (4):  437-442.  doi: 10.16098/j.issn.0529-1356.2018.04.004
    Abstract ( )   PDF (389KB) ( )  

    Objective  To explore the effect of miR-382 on cell proliferation and apoptosis of rat hepatocyte line BRL-3A in vitro. Methods  BRL-3A cells were transiently transfected with miR-382 mimics and inhibitors for 48 hours,MTT assay was used to detect the cell viability. Flow cytometry was used to observe the cell cycle. The expression of proliferation/apoptosis-related genes was detected by Real-time PCR. Results  The expression of miR-382 was significantly up-regulated by treating with miR-382 mimics in the rat BRL-3A cells,and the expression of miR-382 was significantly down-regulated by treating with miR-382 inhibitor in the rat BRL-3A cells(P<0.01). MTT result showed that the activity of BRL-3A cells was significantly increased after miR-382 overexpression. Flow cytometry showed that the number of cells in S phase of miR-382 overexpression group was significantly higher than that of the negative control group, while the number of cells in S phase of miR-382 interference group was significantly lower than that of the negative control group. The mRNA expression levels of poliferating cell nuclear antigen(PCNA), Bcl-2 and Ccnd1 in BRL-3A cells was up-regulated and the mRNA expression levels of Caspase-3 and Bax were down-regulated by Real-time PCR after miR-382 overexpression, while the miR-382 interference group was opposite to the above result. Conclusion  sMiR-382 may promote cell proliferation via regulating the expression of proliferation-related apoptosis-related proteins in rat hepatocyte line BRL-3A.

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    Cancer Biology
    Expression of Golgi phosphoprotein 3 in colorectal cancer tissues and its effects on proliferation, invasion and migration of colorectal cancer cells
    LIU Shi-hong WANG Wei MU Xiao-song
    2018, 49 (4):  461-468.  doi: 10.16098/j.issn.0529-1356.2018.04.008
    Abstract ( )   PDF (744KB) ( )  

    Objective  To analyze the expression of Golgi phosphoprotein 3 (GOLPH3) in colorectal cancer tissues and its influence on the proliferation, invasion and migration of colorectal cancer cells. Methods  The colorectal cancer tissues and paracancerous tissues were collected from the surgically resected colorectal cancer patients (40 cases) without radiotherapy or chemotherapy before the operation. The expression of GOLPH3 in the colorectal cancer tissues and paracancerous tissues were detected by immunohistochemistry, and the relationship between GOLPH3 and clinicopathological features of colorectal cancer was analyzed. Colorectal cancer cell starin with GOLPH3 stable silent was constructed, and a negative control group and blank control group were established. The expression of GOLPH3 protein of cells in the three groups was detected by Western blotting. MTT assay was used to detect the proliferative activity of cells in the three groups. The invasion ability and migration ability of cells in the three groups were detected by transwell test. Results  The expression of GOLPH3 was mainly positive in the cancer tissue sample, and the positive expression rate (65.0%) of the cancer tissues was significantly higher than that of the paracancerous tissues (35.0%) (P<0.05). The GOLPH3 positive expression rate of cancer tissues with TNM stage Ⅲ-Ⅳ, invasion depth≥2 cm, middle-low differentiation and lymphatic metastasis were significantly higher than that of cancer tissues with TNM stageⅠ-Ⅱ, invasion depth<2 cm and high differentiation and without lymphatic metastasis (P<0.05). Compared with the negative control group and the blank control group, the expression level of GOLPH3, the cell proliferation activity, the invasion ability and migration ability rate in the gene silencing group were significantly lower (P<0.05). Conclusion  GOLPH3 is highly expressed in the colorectal cancer tissues, and the expression level is related to the tumor staging, invasion depth, differentiation and lymphatic metastasis. Inhibiting the expression of GOLPH3 gene could deregulate the GOLPH3 protein level, and inhibit the proliferation, invasion and migration of colorectal cancer cells.

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    Effect of camptothecin on the proliferation and apoptosis of HeLa cells
    ZHAO Xing-yu ZHAO Rong TIAN Feng-yu HOU Jian-cheng ZHANG Wei
    2018, 49 (4):  450-454.  doi: 10.16098/j.issn.0529-1356.2018.04.006
    Abstract ( )   PDF (517KB) ( )  

    Objective  To study the effect of different concentrations of camptothecin(CPT) on the apoptosis of human cervical cancer cells (HeLa), and to explore the mechanism of apoptosis induced by CPT in HeLa cells. Methods  The cultured in vitro human cervical cancer cells (HeLa) were divided into the control group and CPT groups treated with various concentrations(100 nmol/L-10 μmol/L). The viability of HeLa cells were detected by methyl thiazolyl tetrazolium (MTT) assay. The morphology changes of HeLa cells were observed with an inverted microscope. The cells apoptosis was analyzed by 4’6-diamidino-2-phenylindole(DAPI) and flow cytometry (FCM). Western blotting was used to detect the expression of Bcl-2, Bax,Cleave-Caspase-3,Cyt-C and poly ADP-ribose polymerase(PARP). Results  Compared with the control group, the viability of HeLa cells decreased after treatment with different concentrations of CPT for 24 hours, and the cells morphology was changed in a dose-dependent manner. DAPI dying showed that there were apoptotic bodies FCM assay indicated that the early apoptosis ratios was increased after treatment with different concentrations of CPT for 24 hours. The cell apoptotic rate increased obviously in the CPT-treated groups than in the control group. The expression of Bcl-2 was decreased whereas the expressions of Bax,cleave-Caspase-3, Cyt-C into cell plasm and 89 kD phosphoribosyl pyrophosphate(PRPP) were increased when compared to that in the control group. Conclusion  CPT can induce HeLa cell apoptosis, and its mechanism may be to activate the mitochondrial dependent pathway, and further activate substrate PARP.

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    Histology,Embryology and Developmental Biology
    Epithelial-mesenchymal transition during the development of pulmonary endoderm-associated second heart field in mouse embryos
    SHI Liang LI Hui-chao JING Ya XIE Jian-shan CHEN Hao CAI Yu-jin YANG Yan-ping
    2018, 49 (4):  480-485.  doi: 10.16098/j.issn.0529-1356.2018.04.011
    Abstract ( )   PDF (702KB) ( )  

    Objective  To explore the relationship between epithelial-mesenchymal transition (EMT) andthe development of pulmonary endoderm-associated second heart field in mouse embryos. Methods  The transforming growth factor-β(TGF-β) 2 and TGF-β3 expressions in mouse embryonic hearts from embryonic day (ED) 9 to ED14 were detected by Western blotting. Both the immunohistochemical and immunofluorescent staining method were used to reveal forkhead box A2 (Foxa2), islet-1 (ISL-1), laminin (LN) and E-cadherin (E-cad) expressions in serial sections of 35 mouse embryos from ED10 to ED12. Results  From ED10 to ED12, both TGF-β2 and TGF-β3 expressed in mouse embryonic heart. In the pulmonary endoderm-associated second heart field development between ED10 and ED12, there was no clear boundary between some ISL-1 positive pulmonary endoderm and surrounding ISL-1 positive second heart field mesenchym which seemed to be intermingled. At the same time, the basement membrane underlying the pulmonary endoderm became interrupted or blurred. Especially at ED11 and ED12, some pulmonary endoderm or its solid cord cells lost Foxa2 expression and showed strongly ISL-1 positive, accompanied by their basement membrane interrupting or missing, E-cad expression decreasing, obvious intercellular space emerging and detachment from endoderm, which suggested the loss of apico-basal polarity. Conclusion  Epithelial-mesenchymal transition is involved in the development of pulmonary endoderm-associated second heart field of mouse embryos.

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    Histological characteristics of the sweat glands and sebaceous glands in adult yaks
    CAO Mi-xia CUI Yan HE Jun-feng YANG Xue ZHANG Qian LIU Peng-gang
    2018, 49 (4):  486-491.  doi: 10.16098/j.issn.0529-1356.2018.04.012
    Abstract ( )   PDF (559KB) ( )  

    Objective   To investigate the histological features of sweat glands and sebaceous glands in Qinghai healthy adult yak. Methods  The histological structures of sweat glands and sebaceous glands in the back, ribs and abdomen of 10 healthy adult yaks were observed and analyzed by HE, periodic acid Schiff(PAS)and Alcian blue periodic acid Schiff(AB-PAS)staining method. Results  The sweat glands of Qinghai adult yak were single tubular-shaped glands, which belonged to the apocrine glands. The cytoplasm of the gland cells contained glycogen. The secretions of sweat glands were acidic and neutral mixtures. Sebaceous glands were botryoid, multilobular, branched acinar glands, which belonged to the holocrine glands. Its secretions were neutral and the cytoplasm of gland cells contained a small amount of glycogen. The density, diameter, length and volume of sweat glands were decreased from the back to the abdomen, but the depth of the skin surface was increased, and the difference was significant (P<0.05). The density, diameter, length and depth of the sebaceous glands were decreased from the back to the abdomen. There was a significant difference between body regions (P<0.05). Conclusion  The morphological structures of sweat glands and sebaceous glands in different body regions of Qinghai adult yak are similar, but its density, size and depth are significantly different, and there is a gradual trend from the back to the abdomen. Compared with other cattles, Qinghai adult yaks have a high density of sweat glands and sebaceous glands, but the function of the glands of Qinghai adult yaks is more developed, which may be related to its adaptation to the plateau alpine environment.

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    Anatomy
    Using the nerve entry point as the blocking target for the subscapularis muscle spasticity
    TIAN Xu-feng HU Shuang-jiang YANG Sheng-bo
    2018, 49 (4):  475-479.  doi: 10.16098/j.issn.0529-1356.2018.04.010
    Abstract ( )   PDF (530KB) ( )  

    Objective This study is aimed to accurately localize the nerve entry point (NEP) of the subscapalaris muscle, and to provide guidance for implementing the chemical neurolysis of the ethanol or phenol injection for the treatment of subscapalaris muscle spasticity. Methods Twenty Chinese adult cadavers in the supine position were examined. The joining lines from the most inferior point of the jugular notch to the apex of the acromion and from the most inferior point of the jugular notch to the junction of sternal body and xiphoid process were designated as the horizontal (H line) and the longitudinal (L line) reference lines of the NEP, respectively. The cadavers were dissected to expose the NEPs of muscular branches of subscapalaris muscle.The NEPs were marked by barium sulfate. Spiral CT scanning and three-dimensional reconstruction were performed. The body surface projection points (P) of the NEPs were determined under Syngo system, P by N projecting to the back skin was designated as P’. The intersections after the vertical P and the H line, after the horizontal P and L line were designated as PH and PL, respectively. The percentage location of PH and PL points on H line and L line and depth of NEPs were measured respectively. Results The points PH of the upper branch and lower branch of subscapular muscle were located at (78.49±5.48)% and (86.17±1.68)% of the H line, and at (21.94±3.59)% and (30.00±1.79)% of the L line, respectively. The depth of NEPs was located at (46.65±2.74)% or (45.87±2.39)%, respectively. Conclusion These results provide guidance for improving the efficacy and efficiency of the chemical neurolysis of the subscapalaris muscle spasticity.

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    Histology,Embryology and Developmental Biology
    Effects of isoliquiritigenin on pulmonary vascular remodeling in chronic hypoxia rat model
    ZHANG Shan-qiang LI Xue-mei YAO Li-jie GUO Lin-na JIANG Yang JIN Hai-feng
    2018, 49 (4):  492-496.  doi: 10.16098/j.issn.0529-1356.2018.04.013
    Abstract ( )   PDF (429KB) ( )  

    Objective  To investigate the possible effect and the underlying mechanism of isoliquiritigenin (ISL) on hypoxic pulmonary hypertension(HPH)through observation of the changes of pulmonary artery pressure, vascular remodeling, and right ventricular hypertrophy in a chronic hypoxia rat model. Methods  Thirty male Sprague-Dawley rats were randomly divided into 3 groups (10 rats per group): normoxia group, hypoxia group and hypoxia group treated with ISL. For hypoxia group treated with ISL, rats were injected intraperitoneally ISL[10 mg/(kg·day), dissolved in 0.5% DMSO]. For normoxia and hypoxia groups, rats were injected intraperitoneally equal amount of 0.5% DMSO. After 4 weeks hypoxia exposure, the right ventricle systolic pressure (RVSP) was recorded using Power Lab Software. The weight ratio of [right ventricle (RV) / left ventricle +septum (LV+S) ] was calculated as an index of RV hypertrophy. After HE staining, the percent medial wall thickness (WT%) and percent medial wall area (WA%) in small pulmonary arteries were determined. The content of superoxide dismutase (SOD) and malonaldehyde (MDA) were measured using commercial kits. NADPH oxidase 4(NOX4) mRNA levels in lung tissues were measured by Real-time PCR. Results  The average RVSP, ratio of RV/LV+S, WT%, and WA% of hypoxia group were increased significantly compared with the normoxia group (P<0.01). However, the average RVSP, ratio of RV/LV+S, WT%, and WA% of hypoxia treated with ISL group were much lower than those of hypoxia group (P<0.01). In addition, the result showed that hypoxia decreased the level of SOD, companied with an increased level of MDA both in lung tissue and in serum (P<0.01). ISL treatment elevated the level of SOD and reduced the level of MDA (P<0.01). Real-time PCR result showed ISL obviously down-regulated the NOX4 mRNA levels in lung tissues (P<0.01). Conclusion  ISL may have beneficial effects on the HPH and these effects may be related to the inhibition of the oxidative stress caused by hypoxia.

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    Treatment and mechanism in total flavonoids of Bidens on adjuvant arthritis in rats
    WANG Ling ZHOU Yu YUAN Yi CHEN Zhi ZHANG Chuan-zhi CAI Ming
    2018, 49 (4):  506-511.  doi: 10.16098/j.issn.0529-1356.2018.04.015
    Abstract ( )   PDF (263KB) ( )  

    Objective  To investigate the effects of the total flavonoids of Bidens (TFB) on the joint swelling index, and the level of inflammatory factors, the toll-like receptor 4(TLR4) signal transduction pathway and the pathological score in the rats with adjuvant arthritis (AA). Methods  Totally 50 male SD rats were randomly divided into blank group, model group, TFB (40 mg/kg, 80 mg/kg, 160 mg/kg) group, each group was 10. The swelling index of joint in each group was compared, and the level of serum interleukin-1β (IL-1β) and tumor necrosis factor α (TNF-α) was detected by radioimmunoassay. Western blotting was used to detect the expression of toll-like receptor 4(TLR4) and nuclear factor-kappa B (NF-κB) in macrophages, and HE staining was used to evaluate the pathological score of rat joints. Results  The joint swelling index, the level of inflammatory factors, the expression of TLR4 and NF-κB in the model group were significantly higher than that in the blank group. In different doses of TFB group (40 mg/kg, 80 mg/kg; 160 mg/kg), the joint swelling index, inflammatory factor level, TLR4, NF-κB expression and pathological score were significantly lower than those in the model group (P<0.05), and the higher the dose of TFB, the lower the expression level of all the indexes. Conclusion  TFB has therapeutic effects on AA rats, and its mechanism may be related to reducing the joint swelling index and inflammatory factor level, inhibiting the expression of TLR4 and NF-kappa B, and preventing pannus formation.

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    Expression and age-related changes of centromere A in rat heart
    FENG Ying YIN Guo-tian LI Ai-rong GUO Zhi-kun
    2018, 49 (4):  512-517.  doi: 10.16098/j.issn.0529-1356.2018.04.016
    Abstract ( )   PDF (1078KB) ( )  

    Objective  To study the expression and age-related changes of centromere protein A(CENP-A) in the rat heart. Methods  Ten rats in each group were collected from postnatal 1 day, 1 week old, 2 weeks old, 3 weeks old, 4 weeks old, 3 months old, 6 months old. The expression and age-related changes of CENP-A in the rat heart were observed by the immunohistochemistry, immunofluorescence, Western blotting, and Real-time PCR techniques. The CENP-A proteins of seven groups were quantitated by SPSS 20.0 statistical software. Results  CENP-A was mainly expressed in the vascular wall, epicardium and epicardial tissue and in a very few cardiomyocytes. The CENP-A in vascular wall was of stable expression with age. But the expression of CENP-A in the epicardium and subepicardial tissues was gradually reduced, which showed that postnatal 1 day was the most and 6 months old are the least. CENP-A in the vascular wall was mainly expressed in the smooth muscle cells, and was basically unchanged with ages.A small number of cardiac myocytes near the subepicardium also express CENP-A, and decreased with age. Conclusion  The expression of CENP-A in the rat heart gradually decreased with aging, but in the vascular wall remained stable expression.

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    Expression and significance of Smad2/3 and Smad7 in follicular cells of chemotherapy-induced premature ovarian failure in rats
    DING Yan TAO Ran ZHANG-Jing CHEN Jing REN Jun-xu
    2018, 49 (4):  497-505.  doi: 10.16098/j.issn.0529-1356.2018.04.014
    Abstract ( )   PDF (1998KB) ( )  

    Objective  To study the effect of cisplatin on ovarian structure and function in rats and its possible molecular mechanism. Methods  Three-month-old SD female rats were divided into control group (C group, n=5) and chemotherapy group (H group, n=5). Rats in H group were given intraperitoneal injection of cisplatin 2 mg/(kg.d), rats in C group were given intraperitoneal injection of equal volume of saline. After 7 days continuous injection, rat ovarian index (ovarian wet weight / body weight) was measured. The levels of serum estradiol (E2) and follicle stimulating hormone (FSH) were tested by enzyme-linked immunosorbent assay (ELISA). The follicle development was observed by hematoxylin-eosin staining. The expression of Smad2, p-Smad2, Smad3, p-Smad3, Smad4 and Smad7 proteins/mRNAs in ovary tissue were detected by immunohistochemistry and Real-time PCR. Results  Compared with C group, the ovarian volume in H group was decreased and the number of growing follicles was significantly decreased (P<0.05). The ovarian index decreased (P<0.05), and the levels of E2 and FSH in serum increased (P<0.05). Immunohistochemistry and Real-time PCR showed that Smad2 (p-Smad2), Smad3 (p-Smad3), Smad4 and Smad7 were expressed in ovarian follicles at all levels, and the expression of Smad2 protein and mRNA in H group increased (P<0.05). The expressions of Smad3, Smad4 and Smad7 protein and mRNA were decreased (P<0.05), and the expression of P-Smad3 was decreased (P<0.05). Conclusion  Cisplatin-induced follicle injury in rats promotes premature ovarian failure, which result in the increase of Smad2 expression, and decrease of Smad3 and Smad7 expression in the follicle cells. The Smad intracellular signal pathway is involved in the process of chemotherapy ovarian injury.

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    Neurobiology
    Pro-apoptosis effects of docosahexaenoic acid on rat C6 glioma cells in vitro
    LI Hao-ming PENG Min YANG Qing-qing YU Ting-ting QIN Jian-bing HAN Xiao JIN Guo-hua
    2018, 49 (4):  419-423.  doi: 10.16098/j.issn.0529-1356.2018.04.001
    Abstract ( )   PDF (414KB) ( )  

    Objective  To clarify the pro-apoptotic effects of docosahexaenoicacid (DHA) on rat C6 glioma cells in vitro. Methods  After treatment of 10, 25, 50, 75 and 100 μmol/L DHA for 24, 48 and 72 hours, C6 cell viability was detected. After treatment of 100 μmol/L DHA for 24 hours, apoptotic cells were labeled by the terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)method, and the proportion of apoptotic cells was counted by flow cytometry. The expression of cleaved Caspase-3 in apoptotic cells was detected by immunocytochemistry and Western blotting. Results  25, 50, 75 and 100 μmol/L DHA inhibited the C6 cell viability at 24, 48 and 72 hours. The TUNEL positive cells were found after treatment of 100 μmol/L DHA for 24 hours. Flow cytometry analysis showed that the proportion of apoptotic cells was increased. Moreover, after treatment of 100 μmol/L DHA, cleaved Caspase-3 positive cells were found by immunocytochemistry. Cleaved Caspase-3 positive protein band was investigated by Western blotting. Conclusion  DHA may induce the apoptosis of C6 cells in vitro.

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    Effects of intervention of brain lipid binding protein expression on differentiation of neural stem cells
    YU Ting-ting LI Hao-ming JIN Guo-hua HAN Xiao QIN Jian-bing
    2018, 49 (4):  424-430.  doi: 10.16098/j.issn.0529-1356.2018.04.002
    Abstract ( )   PDF (878KB) ( )  

    Objective  To explore the effect of BLBP on differentiation of hippocampal neural stem cells (NSCs) into neurons in vitro. Methods  BLBP overexpression adenovirus and BLBP RNA interference lentiviral vector were built. NSCs were obtained from rat hippocampus by serum-free culturing and immunofluorescence was used to identify NSCs. The NSCs were assigned to Ad-NC group, Ad-BLBP group, LV-NC-RNAi group and LV-BLBP-RNAi group. On the 4th day after infection, BLBP mRNA and protein expression in the NSCs were detected by Real-time PCR and Western blotting, and the neuronal class Ⅲ β-tubulin (Tuj1) positive cells were detected by immunofluorescence. Results  NSCs in Ad-BLBP group expressed a high level of BLBP and there were only fewer Tuj1 positive cells than the Ad-NC group; NSCs in LV-BLBP-RNAi group expressed a lower level of BLBP and there were more Tuj1 positive cells than the LV-NC-RNAi group. Conclusion  Down regulation of BLBP expression can promote the differentiation of NSCs into neurons in vitro.

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    Value of different coma scales and 95% spectral edge frequency in predicting one-month mortality of patients with acute coma
    CHEN Xin YAN Xiao-ming KE Kai-fu
    2018, 49 (4):  431-436.  doi: 10.16098/j.issn.0529-1356.2018.04.003
    Abstract ( )   PDF (257KB) ( )  

    Objective  To evaluate the value of different coma scales and 95% spectral edge frequency(SEF) in the central area in predicting the recent death in patients with acute coma. Methods  Fifty-two patients with acute coma admitted in the neurointensive care unit in Affiliated Hospital of Nantong University from August 2014 to October 2016 were included. Glasgow coma scale (GCS), Glasgow-Pittsburgh coma scale (GCS-P), and full outline of unresponsiveness score (FOUR) were performed within 72 hours of the onset of the patients. EEG monitoring was performed and the data of 95% SEF in the central area were recorded simultaneously. The patients were divided into two groups: survival group and death group according to one-month followup. The age, sex, previous history, GCS score, GCS-P score, FOUR score and 95% SEF in the central area were compared between two groups. Single factor and multivariate logistic regression analysis were used to investigate relevant factors related to the recent death. The predictive value of GCS score, GCS-P score, FOUR score and 95% SEF in the central area were compared with the receiver operating characteristic (ROC) curve. The GCS score, the GCS-P score, and the FOUR score combined with the 95% SEF in the central area were compared with the above scores used alone by McNemarχ2 test in the sensitivity and specificity of predicting the recent death. Results   Of the 52 patients, 39 of them were in the survival group and 13 of them were in the death group. Compared with the survival group, the GCS score, GCS-P score, FOUR score and 95% SEF in the central area were significantly decreased in the death group (P<0.05, P<0.01, P<0.01, P<0.01, respectively). Multivariate logistic regression analysis showed that GCS score, GCS-P score, FOUR score and 95% SEF in the central area were independent predictors of one-month death. ROC curve analysis showed that GCS score, GCS-P score, FOUR score and 95% SEF in the central area had moderate predictive value for one-month death. There was no significant difference in sensitivities between the GCS score, the GCS-P score, the FOUR score combined with the 95% SEF in the central area vs the above scores used alone (P>0.05, P>0.05, P>0.05, respectively), but the specificities were significantly higher (P<0.05, P<0.05, P<0.05, respectively). Conclusion  GCS score, GCS-P score, FOUR score, and 95% SEF in the central area can be used to predict the recent death of patients with acute coma. Coma scales combined with 95% SEF in the central area are more effective in predicting the recent death of patients with acute coma.

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    Anthropology
    Estimating body stature by cephalic-facial measurements in Tujia ethnicity
    YU Ke-li WEI Yu ZHANG Xing-hua ZHAO Da-peng ZHENG Lian-bin WANG Wen-jia
    2018, 49 (4):  518-523.  doi: 10.16098/j.issn.0529-1356.2018.04.017
    Abstract ( )   PDF (231KB) ( )  

    Objective  To conduct stature estimation from seven facial measurements of 1901 Tujia adults (858 males, 1043 females) in Southern China aged from 18 to 75 years. Methods  The present study applied variable statistical method, such as multiplication factors, linear regression, multiple regression as well as multiple stepwise regression to explore the relationship between stature and cephalic-facial measurements and the reliability of the estimation method of stature in Tujia. Results  All the facial measurements had significant coefficient with stature according to Pearson’s analysis. Linear regression was the best method for stature estimation according to cephalic-facial measurements in Tujia among the four method. Overall, the range of the four method based on the reliability was linear regression, multiple regression, multiple stepwise regression and multiplication factors. Conclusion  Cephalic-facial measurements can be used to establish stature estimation equation. Horizontal head circumference has the highest correlation in the present study so that the estimating equation based on horizontal head circumference is most accurate.

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    Polymorphism of leptin gene G-2548 A and pregnancy-induced hypertension syndrome in Han population of Qinghai Province
    ZHUANG Wen-ting ZHAO De-xiong WANG Xiang-lin LI Hong-rong LI Chang-xing LI Fu-xin KONG De-xia LI Jian-hua
    2018, 49 (4):  524-528.  doi: 10.16098/j.issn.0529-1356.2018.04.018
    Abstract ( )   PDF (266KB) ( )  

    Objective To investigate the relationship between polymorphism of leptin gene promoter region G-2548 A and pregnancy-induced hypertension syndrome(PIH)in Han population of Qinghai Province. Methods  A total of 130 PIH subjects and 135 normal pregnancy subjects were studied. The genotype of leptin was analyzed by polymerase chain reaction fragment length polymorphism (PCR-RFLP). The mutation was confirmed by sequencing. Results  The frequencies of leptin GG, GA, and AA genotype in the PIH group were 53.1%, 40% and 6.9%, and in the control group were 60%, 34.1% and 5.9% respectively. There was difference in frequency distribution of leptin genotype between the PIH and control groups. The frequency of G gene in the PIH group was higher than the control group (χ2=4.21,P<0.05,OR=0.66,95%CI=0.45-0.98). Conclusion  Leptin gene G-2548 A is significantly correlated with PIH in Han group of QingHai. G allele may be a susceptible gene to PIH (OR=1.361, 95%CI: 1.01-1.83) and A allele may be a protective gene to PIH (OR=0.901, 95%CI: 0.82-1.00).

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    Comparison of mandible measurements during Holocene
    LI Hai-jun FAN Xiao-xia FANG Lu-yu DAI Cheng-ping
    2018, 49 (4):  535-539.  doi: 10.16098/j.issn.0529-1356.2018.04.020
    Abstract ( )   PDF (456KB) ( )  

    Objective   To analyze the time variation of gender size difference rates of human mandibles in Holocene, and to explore the change rates of the mandible sizes of males and females among different time periods. Methods  Using 520 mandibles from several archaeological sites in China, we used SPSS 18.0 to compare the metric traits between the male and female in the same and different time periods, by independent sample t test, and calculate the gender difference rates and the change rates among different time periods. Results  The sizes of male mental foramen and mandibular foramen in present mandibles were bigger than that in Neolithic and Bronze-Iron Age, while the difference of the female among different time periods was not significant. The gender difference rate of the size of mental foramen and mandibular foramen was bigger in present than that in Neolithic and Bronze-Iron Age. Conclusion  The gender difference rates in mandible are greater in recent population than that in Neolithic and Bronze-Iron age. The comparison of cross section perimeter ratio indicates that the robust of mandible body at mandibular first molar(M1) declines compared with that at mandibular first premolar(P3) in Holocene. The position of mental foramen became higher during Neolithic to present.

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    Size variation and related implications of Chinese female mandibles in Holocene
    LI Hai-jun CHEN Feng ZHANG Yi-dan DAI Cheng-ping
    2018, 49 (4):  529-534.  doi: 10.16098/j.issn.0529-1356.2018.04.019
    Abstract ( )   PDF (238KB) ( )  

    Objective   To better understand variation of Holocene Chinese mandibular morphology, a study was conducted on 215 adult female mandibles. Methods  In this study, 23 mandible measurement traits were measured using traditional anthropometric method. SPSS 18.0 software was used to carry out the average calculation of measurement trait data of each era and the significance test among different periods. Results  The overall size of female mandibles decreased from the Neolithic to Bronze-Iron Ages to present day. The linear characters of the female mandiblular features varied between different time periods. The decrease of thickness of the mandibular corpus primarily occurred during the Neolithic to Bronze-Iron Ages. The decrease in main size and body height was during the Bronze-Iron Ages to present day. It is possible that mandibles became thinner before the overall size decreased. Conclusion  The comparison shows that the evolvement trend of the mandibular of the Holocene female is basically consistent with that of male.

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    Body constitution of Chinese Qiang adults
    DU Hui-min SUN Ze-yang ZHANG Xing-hua YU Ke-li GAO Wen-fang XU Bo-song GAO Yuan WEI Yu ZHENG Lian-bin
    2018, 49 (4):  540-542.  doi: 10.16098/j.issn.0529-1356.2018.04.021
    Abstract ( )   PDF (152KB) ( )  

    Objective  To collect body composition data of Qiang adults in Maoxian, Sichuan Province, in order to reveal the body composition characteristics of Qiang. Methods  A total of 602 adults of Qiang were randomly selected. The bioelectrical impedance method was used to measure the body composition of the subjects. The data were processed by SPSS 17.0. Results  There was a statistically significant difference between male and female Qiang adults in all index. The index related to fat in Qiang adults had a significant correlation with age. Percent body fat, percent trunk fat and visceral fat level increased gradually with age. The total body muscle mass increased first and then decreased with the increase of age. 58.6% of males and 63.7% of females were overweight or obese, 31.8% of men and 43.6% of women were at central obesity, which may lead to a higher prevalence of obesity-related diseases. Conclusion  The Qiang adults have more body fat and more developed muscles and bones, which may be related to the low temperature in Maoxian.

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    Review
    Regulation of neuroimmune system on learning and memory
    DING Yan-ping JIN Yi-ran BAI Yu-jie PAN Zi-yan SHAO Bao-ping
    2018, 49 (4):  543-548.  doi: 10.16098/j.issn.0529-1356.2018.04.022
    Abstract ( )   PDF (244KB) ( )  

    Nervous system and immune system are two main controlling systems for sustaining internal balance of the body, which can mutual regulate each other. The nervous system regulates the immune response of the organism through the nervous and endocrine pathways by releases various cytokines, transmitters and neuropeptides. In turn, immune modulator or neural active substances produced by immune system can regulate the functions of the nervous system. The interaction between the nervous system and the immune system plays an important role in the maintenance and development of the normal function of the nervous system. We review research progress of roles of the immune system include microglia, inflammatory cytokines, especially interleukin (IL)-1β, IL-6, tumor necrosis factor and Toll-like receptors, and peripheral immune system in the normal hippocampal functions such as learning and memory, synaptic transmission, and nerogenesis.

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    Role of Nogo-A and Nogo-A receptors in Alzheimer’s disease
    WANG Zhao-lun ZHANG Yan WANG Jun
    2018, 49 (4):  549-555.  doi: 10.16098/j.issn.0529-1356.2018.04.023
    Abstract ( )   PDF (353KB) ( )  

    Nogo-A is an important myelin associated inhibitory factor, which creates a critical barrier to the central nervous system regeneration after injury. Recent research shows that Nogo-A/Nogo-A receptors can affect the metabolism of amyloid-β-protein (Aβ), and its downstream ROCK kinase. It can modulate the level of Aβ and tau protein, as well as the permeability of blood brain barrier. These findings suggest that Nogo-A/Nogo-A receptors have a close relationship with Alzheimer’s disease (AD). Moreover, two new receptors of Nogo-A, PirB and S1PR2, have been identified recently. The identification of these two receptors may provide new insight into the mechanism of Nogo-A involving in the progress of central nervous diseases. we focuses on the up-to-date knowledge of the basic structure and function of Nogo-A/Nogo-A receptors and their role in AD pathogenesis.

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    Research progress of the role of microglia in the brain injury after subarachnoid hemorrhage
    PENG Shu-chen YANG Xiao-mei
    2018, 49 (4):  556-560.  doi: 10.16098/j.issn.0529-1356.2018.04,024
    Abstract ( )   PDF (212KB) ( )  

    Microglia, a member of mononuclear phagocyte system, plays a significant role in maintaining homeostasis in central nervous system(CNS). As a subtype of stroke, subarachnoid hemorrhage (SAH)leads to an early brain injury within 72 hours and a delayed brain injury in 3-5 days. Vasospasm used to be regarded as a crucial mechanism of brain injury after subarachnoid hemorrhage, but recently increasing attention is focused on inflammation in brain injury. Microglia polarize to M1 type to act as an pro-inflammatory agent or to M2 type to become a neuro-protector in inflammation in both early stage and delay stage after subarachnoid hemorrhage. Here, we summarize researches on the role of microglia in the brain injury after subarachnoid hemorrhage, to find the research hotspot and provide potential targets for further clinical investigations.

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