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    Cell and Molecules Biology
    Cardiac fibroblasts of the rat transfected by lenti virus-stromal cell-derived factor-1α-green fluorescent protein vector
    ZHU Zhan-zhan HOU Bin SUN Wen-wen YUAN Hui-fang ZHANG Yong-chun CAI Xin-hua
    2017, 48 (1):  43-47.  doi: 10.16098/j.issn.0529-1356.2017.01.008
    Abstract ( )   PDF (612KB) ( )  

    Objective To investigate the effect of lentivirus-stromal cell-derived factor-1α-green fluorescent protein(LV-SDF-1α-GFP) on the cardiac fibroblasts, the optimum conditions of infection, the expression and secretion of the target protein. Methods The cardiac fibroblasts of neonatal rats were primarily isolated and cultured by differential adherence methods, and were observed and identifi with immunofluorescence. LV-SDF-1α-GFP with different titers and conditions was transfected into cardiac fibroblasts. The expression of fluorescence and the optimal transfection conditions were observed. LV-SDF-1α-GFP target gene virus and negative control CON145 virus were transfected into cardiac fibroblasts. The growth curve was drawn, and the effect of transfection on the proliferation of cardiac fibroblasts was explored.The cardiac fibroblasts were transfected with the optimum transfection dose, and the expression of SDF-1α was detected by Dot-blotting. The measurement data underwent statistical analysis. Results There was no statistical difference between the cardiac fibroblasts with SDF-1α transfected lentivirus and without no-transfected SDF-1α lentivirus. The peak of the expression of SDF-1α appeared in culture day 4 and statistical analysis showed significantly difference (P<0.05). Conclusion The LV-SDF-1α-GFP vector is of higher transfection efficiency to cardiac fibroblasts with the both low cytotoxicity and ability of secreting SDF-1α protein.

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    Screening of candidate molecules interacting with protein kinase Pkmyt1 using yeast two hybrid technique
    LIU Chao MENG Zhi-chao LUAN Zhi-dong REN Li-li LIU Yi-meng XIAO Jian-ying
    2017, 48 (1):  48-53.  doi: 10.16098/j.issn.0529-1356.2017.01.009
    Abstract ( )   PDF (475KB) ( )  

    Objective Protein kinase Pkmyt1 negatively regulates the mitosis of mouse 1-cell stage fertilized eggs, but the specific mechanism is not clear. Therefore, the yeast two hybrid system was used to screen the candidate proteins interacting with Pkmyt1 from human ovary cDNA library in order to provide a new clue for the study of Pkmyt1 regulating the early development of mouse fertilized eggs. Methods Mice ovarian tissue cDNAs was used as a template to build pGBKT7-Pkmyt1 bait plasmid, and pGBKT7-Pkmyt1 was transformed into yeast competent cells at SD/Trp(SDO), SD/Trp/X-α-Gal(SDO/X) and SD/Trp/X-α-Gal/AbA plates(SDO/X/A) medium to detect the toxicity and self-activation ability of yeast and detect its expression in yeast using Western blotting method. The pGBKT7-Pkmyt1 containing yeast cells was fused with human ovary cDNA library, followed by screening positive clones by PCR, plasmid extraction, DNA sequencing and detection the yeast self-activated ability again, the use of bioinformatics analysis of selected proteins and embryo development. Results The enzyme digestion analysis and Blast analysis showed that the pGBKT7-Pkmyt1 plasmid was successfully constructed. When the plasmid was transferred into golden Y2H, the SDO medium was evenly grown, and no clones were grown on the SDO/X/A plate, and the protein was detected by Western blotting to detect the expression of Pkmyt1 in the yeast cells. After PCR validation, the positive clones were inserted into fragments. 182 proteins which interact with Pkmyt1 were obtained by preliminary screening, and 46 proteins were further verified by the recovery experiment with Pkmyt1. Conclusion Totally 46 proteins were found through the selection of the interaction between Pkmyt1 and they regulated in mouse oocyte maturation and early development of fertilized eggs.

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    Histology,Embryology and Developmental Biology
    Effect of altitude chronic hypoxic on morphology and structure of carotid body in yak and migrated cattle
    LIU Feng-yun MA Lan HU Lin LI Yu-xian LIU Shi-ming WU Tian-yi
    2017, 48 (1):  70-77.  doi: 10.16098/j.issn.0529-1356.2017.01.013
    Abstract ( )   PDF (1039KB) ( )  

    Objective Yak (Bos grunniens) is an indigenous mountain species and genetically adapted to high altitude. When cattle from low altitude is taken to a high altitude, it may develop hypoxic pulmonary artery hypertension known as brisket disease. We compared the morphological and histological variations of carotid body (CB) among yaks, migrated cattle and low altitude cattle. The results could provide important insights of the process of adaptive evolution. Methods The CBs were sampled from 9 indigenous yaks born and lived at 3000-4000 m, and 9 cattle whose ancestors migrated to high altitude at 2500m for several generations in Qinghai-Tibet plateau. In addition, 12 CBs of low altitude cattle living at 1300 m in Gansu were used as control. The samples were fixed, sectioned, stained, observed under an optical and an electron microscope. The expression of hypoxia-inducible factor-1α (HIF-1α), nitric oxide synthase (NOS), leptin receptor (LEPR) and erythropoietin (EPO) were observed by immunohistochemical methods. Results No significant difference was observed for the size of CB in yaks which lived in various altitude. Furthermore, the size, shape, and the percentage of chief cells and sustentacular cells from each CB of yaks had no significant difference either. However, the CB size of migrated cattle was the largest comparing to the lowland cattle and yaks. The CB of yaks was the smallest at size. The percentages of light chief cells, dark cells, pyknotic cells of each CB in yak were 67.1%∶28.2%∶4.7%, and that in migrated cattle were 78.5%∶18.6%∶2.9%, while that in lowland cattle were 87.3%∶10.2%∶2.5%. Similar to CB of lowland cattle, a small amount of clear cells of yak were of clear cytoplasm; and nuclear chromatin of dark cells was denser with more particles. In the CB in migrated cattle, the clear cells were mostly of clear cytoplasm, and few particles were investigated in nuclear chromatin of dark cells, but there were no distinct difference in sustentacular (type 2) cells from CB in these three species. Compared with lowland cattle, a small amount of nucleus heterochromatin of chief cells (type 1) in CB in migrated cattle increased. Some of chief cells exhibited anomalies in morphology. Intracytoplasmic organelles of chief cells were mostly swelling and dissolved, which resulted in small remnant of organelle and dense core vesicles. The expression of HIF-1α, NOS, LEPR and EPO varied in CB in these three species, while the positive expression rate of EPO in CB from yak was below that from lowland cattle(P<0.05). Conclusion High altitude does not cast significant influence on the size of CB and the amount of chief cells in CB of yak. There is a certain extent of convergence in the ratio of different types of chief cells in CB between yak and migrated cattle, but the functional system of the type 1 cells is damaged because of the hypoxia in migrated cattle, suggesting that migrated cattle are experiencing the process of acclimatizing themselves to high altitude. Chronic hypoxia may affect the EPO protein expression in the CB from yak and migrated cattle.

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    Impact of active vitamin D3 on the expression of miR-29a and on the production and development of pulmonary fibrosis in rats
    XU Meng-ting LIU Nai-guo DONG Hong-liang ZHENG Jing NI Na WANG Nan
    2017, 48 (1):  78-86.  doi: 10.16098/j.issn.0529-1356.2017.01.014
    Abstract ( )   PDF (2043KB) ( )  

    Objective To investigate the preventive and therapeutic effects of active vitamin D3[1,25(OH)2D3] on the production and development of pulmonary fibrosis in rats, and the effect on miR-29a. Methods A total of 150 male SD rats were randomly divided into fibrogenic intervention group (n=90) and fibrotic-intervention group (n=60), then subdivided into model group Ⅰ, treatment group Ⅰ and control group Ⅰ(n=30)for fibrogenic intervention group, and model group Ⅱ, treatment group Ⅱ and control group Ⅱ(n=20)for fibrotic-intervention group. Bleomycin(5mg/kg)was injected into the rat trachea to establish the model of pulmonary fibrosis in the model groups Ⅰ/Ⅱ and treatment groups Ⅰ/Ⅱ, while the control groupsⅠ/Ⅱ were injected with isopyknic sterile saline. The treatment groups Ⅰ/Ⅱ were intraperitoneal injected with active vitamin D3 on the 2nd day and 14th day after surgery respectively. The model groupsⅠ/Ⅱ were intraperitoneally injected with solvent of vitamin D3 (0.1% ethanol in 99.9% propylene glycol ), the control groupsⅠ/Ⅱ were injected intraperitoneally with sterile saline, on the 2nd day and 14th day after surgery respectively. All injections were carried out once every other day. In the fibrogenic intervention group, 10 rats were euthanized at 14th, 21st and 28th day in each group, and in the fibrotic-intervention group, 10 rats were euthanized at 21st and 28th day in each group respectively. After obtaining lung tissues from experimental rats, the differences of collagen fiber and hydroxyproline content were compared by the Masson staining and basic-hydrolysis method Respectively. The expression of α-smooth muscle actin (α-SMA), type Ⅰ collagen (ColⅠ) mRNA and miR-29a were detected by Real-time PCR. The protein expressions of α-SMA and Col Ⅰwere measured by immunohistochemical technology, and quantized with image analysis. Results Fibrosis appeared in lungs of experimental rats treated with bleomycin after 14 days, and gradually aggravated with time. At 14th, 21st and 28th day, the hydroxyproline content, the mRNA and protein expression levels of α-SMA and ColⅠin model groupⅠ/Ⅱ and treatment group Ⅰ/Ⅱ were significantly higher than that of control group Ⅰ/Ⅱ, but the expressions of miR-29a in model groupⅠ/Ⅱ and treatment group Ⅰ/Ⅱ were significantly lower than that of control group Ⅰ/Ⅱ. Compared with model groupsⅠ/Ⅱ, the expressions of miR-29a was increased in treatment groups Ⅰ/Ⅱ (P<0.05). In the fibrogenic intervention group, the hydroxyproline content, the mRNA and protein expression of α-SMA and Col Ⅰin treatment group Ⅰwere all obviously reduced at three time points compared with model groupⅠ (P<0.05). In the fibrotic-intervention group, the hydroxyproline content, the mRNA and protein expression of α-SMA and Col Ⅰwere slightly decreased in treatment group Ⅱ compared with model groupⅡ, but no significant difference was found (P>0.05). Conclusion Active vitamin D3 may have an inhibiting effect on genesis and progression of pulmonary fibrosis in rats, with better preventive effect than therapeutic effect, and may promote the expression of miR-29a, suggesting that active vitamin D3 inhibits the genesis and progression of pulmonary fibrosis probably by promoting the expression of miR-29a.

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    Effect of cryopreservation on the expression of sperm surface protein PH-20 and ratio of apoptosis in normal birth and infertility
    MA Xiao-ping GAO Xiao-qin ZHOU Hua WANG Jun
    2017, 48 (1):  87-91.  doi: 10.16098/j.issn.0529-1356.2017.01.015
    Abstract ( )   PDF (383KB) ( )  

    Objective To investigate the effect of cryopreservation on the expression of sperm surface protein PH-20 and the sperm apoptosis. Methods Semen samples were obtained from fertile men (n=14, group A) and infertile men (n=20, group B). Western blotting was used to detect the PH-20 protein expression in human spermatozoa.The localization of this protein on human spermatozoa was determined by indirect immunofluorescent staining using PH-20 antibody. The sperm apoptosis was examined by terminal deoxynucleotidyl transferase(TdT) mediated deoxyuridine triphophate-biotin nick end labeling(TUNEL). Results After cryopreservation, the level of PH-20 protein expression was significantly lower in both group A and B than that of fresh sperm (P<0.05). The percentage of PH-20 positive rate was significantly lower in both group A and B than that of fresh sperm (P<0.05). The ratio of sperm apoptosis was not significantly different in thawed group A than that of fresh sperm (P>0.05). The ratio of sperm apoptosis was significantly lower in thawed group B than that of fresh sperm (P<0.05).
    Conclusion Cryopreservation caused significant reduction of PH-20 protein expression, the percentage of PH-20 positive rate in human sperm. But the cryopreservation had no significant influence on normal fertility of sperm apoptosis.

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    Cancer Biology
    Expression and clinical significance of Ezrin in the serous ovarian cancer
    XIE Zhe LI Lan-fang LUO Qing-ya LIU Yi LI Gui-qin WANG Geng-chao MAO Cheng-yi YI Ping
    2017, 48 (1):  54-60.  doi: 10.16098/j.issn.0529-1356.2017.01.010
    Abstract ( )   PDF (505KB) ( )  

    Objective To investigate the expression feaure of Ezrin in the normal ovarian epithelial tissues and the serous ovarian cancer tissues and evaluate the relationships between Ezrin expression and the clinicopathological characteristics of ovarian cancer patient. Methods The relative Ezrin mRNA- expression was detected by Real-time PCR in 40 specimens of ovarian carcinoma and 27 specimens of normal ovarian epithelial fresh frozen tissues. A total of 134 specimens of paraffin-embeded ovrian cancer tissues and 27 specimens of normal ovarian epithelial tissues were collected,and the Ezrin protein level was examined by immunohistochemistry. The correlation of Ezrin with the clinicopathological characteristics of patients with ovarian cancer was analyzed by SPSS 20.0. Results Both the RNA and protein levels of Ezrin were significantly reduced in ovarian cancer compared to the normal control(P<0.05). Statistical analysis revealed that Ezrin expression had no correlation with age, cytoreduction satisfactory degree and sensitivity of chemotherapy. However there was a significant difference in ezrin protein levels with the disease-free and overall survival (P<0.05). Although ezrin did not serve as an independent predictor for disease progression (HR 1.406, P>0.05) and overall survival (HR 1.260, P>0.05), the expression of Ezrin was negatively correlated with clinical stage,differentiation, and Oentum majus metastasis (P<0.05). Conclusion The down-regulation of Ezrin may play a substantial role in the pathophysiology of serous ovarian cancer. Therefore, it may be used as the potential pre-diagnostic and prognostic marker of ovarian cancer and direct the clinical treatment in the future.

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    Review
    Mechanism of autophagy opens a new way for treatment of diseases
    WANG Hai-jie TAN Yu-zhen
    2017, 48 (1):  103-105.  doi: 10.16098/j.issn.0529-1356.2017.01.018
    Abstract ( )   PDF (143KB) ( )  

    Autophagy summarizes all processes in which intracellular material is degraded within the lysosome and where the macromolecular constituents are recycled. Japanese scientist Yoshinori Ohsumi discovered 15 autophagy-related genes (ATG) and elaborated autophagic mechanisms. He has won the 2016 Nobel Prize in Physiology or Medicine. His pioneering research achievements provide an important base for studying physiological and pathological effects of cell autophagy and open a new way for treatment of diseases by modulating autophagy. Autophagy is a common cellular response. The level of autophagy is low in normal condition, while physiological and pathological stimulations up-regulate autophagy significantly. ATG deletion or autophagic disfunction may cause some diseases. Recently, prevention and treatment of autophagic disfunction-related diseases by activating or inhibiting cell autophagy have been tried.

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    Polarization and functional regulation of macrophage
    GONG Tian-tian HUANG Shao-gang ZHANG Yue LI Jia LEI Lei SHAN Zhi-yan
    2017, 48 (1):  106-110.  doi: 10.16098/j.issn.0529-1356.2017.01.019
    Abstract ( )   PDF (253KB) ( )  

    Macrophages have the great plasticity in various tissues in vivo, which play important roles in the development and homeostasis. In response to certain inductors, macrophages can change their phenotype and result in polarization. Polarized macrophages can react to the immune response and participate in tissue repair and remodeling, which have the significant clinic application value. In this review, the classification, regulation mechanism and reprogramming of polarization are addressed, which may provide a theory basis for research of macrophage polarization.

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    Related research progress of autophagy in cervical cancer cells
    TONG Yan TAN Bu-zhen FANG Yu-ting LAI Zhi-qing WANG Yu WANG Xiao-hong
    2017, 48 (1):  111-114.  doi: 10.16098/j.issn.0529-1356.2017.01.020
    Abstract ( )   PDF (185KB) ( )  

    Cell autophagy refers to use lysosomes to degrade the misfolded proteins and the damaged organelles in the cytoplasm. This process can reuse the intracellular degradation substances in order to maintain stable environment in the cell.Normal cells maintain at a low level of autophagy. When a cell is under the condition of unfavorable factors, its autophagy can be activated against unfavorable factors.Autophagy plays an important role in cancer occurrence,development, metastasis, recurrence and drug resistance, but the detailed mechanism of autophagy in these areas is still unclear.In this paper, the autophagy\| related genes in cervical cancer cells and their application in cervical cancer treatment are reviewed.

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    Anthropology
    Relationship between anterior cruciate ligament anatomical reconstruction femoral tunnel and lateral collateral ligament femoral tunnel
    DONG Yi-long QIAN Yue-nan ZHONG Xi-qiang LIU Liang-le CAI Chun-yuan
    2017, 48 (1):  61-64.  doi: 10.16098/j.issn.0529-1356.2017.01.011
    Abstract ( )   PDF (319KB) ( )  

    Objective To investigate the relationship between the anterior cruciate ligament (ACL) single-bundle anatomical reconstruction femoral tunnel and the lateral collateral ligament(LCL) reconstruction femoral tunnel, in order to provide the anatomical data for ACL and LCL one-stage reconstruction and to avoid mutual interference between the two femoral tunnels. Methods Thirty adult cadaveric knees without degeneration, deformity and joint damage were utilized. The cutting ends of each specimen were at least 20cm above and below the knee. The cadavers were 16 males and 14 females, aged from 23 to 66 years, 38.7 years for average. The femoral tunnel was drilled through an accessory medial portal (AMP) at 120°of knee flexion. The posteriolateral corner ligament (PLC) in the outer surface of lateral femoral condyle was visualized and the LCL was isolated and the LCL tunnel was drilled. All the specimens were underwent CT scan to observe the tunnel collisions on CT slices, and calculate the shortest distance between two tunnels. Results The shortest distance between the LCL reconstruction femoral tunnel and the ACL reconstruction femoral tunnel was (3.90±2.40)mm when the depth of LCL tunnel was 25mm. The shortest distance between the LCL tunnel and the ACL tunnel was (2.70±1.90)mm when the LCL tunnel depth was 30mm. The shortest distance between the LCL tunnel and the ACL tunnel was (4.4±2.60)mm when the ACL tunnel depth was 25mm. The shortest distance between the LCL tunnel and the ACL tunnel was (3.2±2.10)mm when the ACL tunnel depth was 30mm. Among 30 specimens, 6 cases were found tunnel collisions, the collision probability as high as 20%. Conclusion The risk of tunnel collisions between the reconstruction femoral LCL and ACL tunnels is high. Thus, when the one-stage of clinical anatomical reconstruction, preoperative preparation should be individualized, and the length and diameter of the femoral tunnel for reconstruction LCL should be planned to avoid collision with the ACL femoral tunnel.

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    Anatomy
    Anatomy of the scapulae safety prediction
    ZHANG Zhen-hua WANG Li-bo CUI Wen-ming WEN Jing PENG Shi-qin SHI Zi-yu XU Gao-lei
    2017, 48 (1):  65-69.  doi: 10.16098/j.issn.0529-1356.2017.01.012
    Abstract ( )   PDF (278KB) ( )  

    Objective To assess the differences between different scapulae in the same subject to obtain a safe zone for avoiding iatrogenic nerve lesions, and to provide data reference for the improvement and operation of arthroscopic surgery. Methods Totally 100 well preserved dry scapulae were selected and measured the scapular body, glenoid, and the course of the suprascapular nerve. Data were processed and analyzed by statistical methods. Results The Pearson’s correlation between major ongitudinal axis (A), major transversal axis (B), glenoid major longitudinal axis (C), glenoid major transversal axis (D) and the distance from the supraglenoid tubercle to the scapular notch (E) respectively were 0.797, 0.786, 0.792 and 0.687; between them and the distance from the midline of the posterior glenoid rim to the base of the scapular spine (F) respectively were: 0.368, 0.381, 0.403 and 0.396. According to the results of statistical calculation, the safe zone referring to the whole population proved to be 2.2cm for the distance from the supraglenoid tubercle to the nerve at the scapular notch and 1.3 cm for the distance from the midline of the posterior glenoid rim to the base of the scapular spine. Conclusion It is important to understand the safety zone in the operation to avoid the injury of the suprascapular nerve; therefore, the related indexes of the scapulae should be evaluated, so that as obtain the better entry during the operation. The linear predictors should be used to obtain specific values of the posterosuperior limit in each patient.

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    Anthropology
    Comparison between different measurements of the body fat rate for 6-8 years’ bone aged girls
    ZHANG Qing LIU Zhi-guo YANG Tao BAO Juan LI Shan-shan
    2017, 48 (1):  92-98.  doi: 10.16098/j.issn.0529-1356.2017.01.016
    Abstract ( )   PDF (317KB) ( )  

    Objective To compare the different measurement methods of the body fat rate for 6-8 years’ bone aged girls,and establish a formula based on the skinfold measurement. Methods Fifty-nine 6-8 years bone aged girls in primary schools were recruited and divided into three groups which were the underweight,normal and overweight groups. Fat rates were tested respectively by dual-energy X-ray absorptiometry(DEXA), bioelectrical impedance method(BIA), and skinfold method. All the data were statitically analysised with SPSS 13.0. Results There was a significant correlation between the BIA and the DEXA measurements in normal and especially in obese girls. Changling formula significantly overestimated the body fat rate in underweight and normal group. Yao Xing-jia formula significantly underestimated the body fat rate in overweight group.Yuan Tian-heng formula significantly overestimated the body fat rate in all three groups. We established a body fat rate formula based on skinfold measurement: body fat percentage=3.919 +0.715 skinfold in waist circumference+0.592 skinfold in triceps. Conclusion For normal and overweight bone-aged 6-8 years’girls, BIA is an effctive way to measure their body fat rate. Changling,Yuan Tian-heng, and Yao Jia-xings’ fomulas showed some deficency in overestamating or underestimateing their body fat rate. Skinfold of waist circumference together with skinfold of triceps may effectively evaluate their body fat rate.

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    Sexual dimorphism by the length of metacarpals and phalanges of Macaca mulatta
    ZHAO Xiao-jin WANG Feng-chan TIAN Hua-xiang DUO Tian
    2017, 48 (1):  99-102.  doi: 10.16098/j.issn.0529-1356.2017.01.017
    Abstract ( )   PDF (240KB) ( )  

    Objective To investigate the utility of length measurements of the metacarpals and phalanges to estimate sex in a Macaca mulattapopulation. Methods The length of the metacarpals and phalanges of 11 males and 25 females was measured by direct dry bone measurement. Data analyses were performed using SPSS version 20.0. Results The result showed that the accuracies of sex determination of hand bones gradually increased from metacarpal to distal phalange. The average lengths of all five metacarpals and distal phalanges were sexually dimorphic with accuracies of 88.9% and 100.0% respectively. The total lengths of the first hand bone (including one metacarpal and two phalanges) and the fifth hand bone (including one metacarpal and three phalanges) were sexually dimorphic with accuracies of 90.3% and 100.0% respectively. The result according to factor analysis showed that the morphologic classes for all 19 hand bones were divided into the rows (metacarpals, proximal phalanges, middle phalanges, and distal phalanges) rather than the rays (five digits). Conclusion The results of this study suggest that the length of metacarpals and phalanges in monkeys can be used reliably for sexing and be a valuation tool both in theory and application in biological context in primates.

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    Cell and Molecules Biology
    Effect of Heshouwuyin on protein expression of steroidogenic acute regulatory protein and cytochrome P450 cholesterol side chain cleavage enzyme in Leydig cells of rats
    WANG Jian-mingSUN Jing QI Feng SONG Qing-liang NIU Si-yun HUI Chen-hong XU Tian-rong HE Fei
    2017, 48 (1):  30-36.  doi: 10.16098/j.issn.0529-1356.2017.01.006
    Abstract ( )   PDF (599KB) ( )  

    Objective To explore the effect of Heshouwuyin on the steroidogenic acute regulatory protein (StAR) and cytochrome P450 cholesterol side chain cleavage enzyme(P450scc) protein expression in Leydig cells of rats. Methods Juvenile male Wistar rat,10-20 days old, were used to isolate the Leydig cells and cultured, and then collected them. The oxidative damage technique was used to establish Leydig cell aging model. This study had a Leydig aging model, Heshouwuyin and Shouwu pill treatments, and normal and Heshouwuyin control group. Results The intensities of immunohistichemical staining for StAR and P450scc were significantly lower in the Leydig aging model group than those in the normal group and Heshouwuyin control group (P<0.01), and were significantly higher in the Heshouwuyin and Shouwu pill group than that in the aging model group (P<0.05). Heshouwuyin group was better than Shouwu pill group. Conclusion Heshouwuyin can increase the protein expression of StAR and P450scc in Leydig cells, improve the synthesis of testosterone, thus delaying the senescence of Leydig cells.

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    Bioinformatic analysis of gene expression of human peripheral blood cells related to radiation injury
    XU You-qin ZHANG Qing-fen ZHANG Chao
    2017, 48 (1):  37-42.  doi: 10.16098/j.issn.0529-1356.2017.01.007
    Abstract ( )   PDF (445KB) ( )  

    Objective To investigate genes associated with radiation injury and to reveal human peripheral blood cells change during radiation. Methods The microarray data of radiation induced gene expression in human blood were downloaded from the Gene Expression Omnibus (GEO) database and Qlucore Omics Explorer 3.0 software was used to screen differentially expressed genes. Further analysis of differentially expressed genes was conducted by the on-line tools STRING and DAVID. Results Of 94 differentially expressed genes, 31 genes were of co-overexpression and 11 genes were co-underexpressed. These genes were involved in the biological process and molecular function of regulation of apoptosis, regulation of programmed cell death, regulation of cell death, intracellular signaling cascade, response to DNA damage stimulus and regulation of cell cycle. STRING analysis showed that ubiquitin C(UBC),proliferating cell nuclear antigen(PCNA),murine double minute-2(MDM2) had important roles in the protein-protein interaction network, which participates in p53 pathway. Conclusion UBC, PCNA and MDM2 may be potential therapeutic targets of ionizing radiation exposure through the bioinformatics analysis, which needs further study.

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    Review
    Development of chimeric antigen receptor modified T cells and the selection of tumor target antigen
    TANG Hai-jun LIU Yu-qin
    2017, 48 (1):  115-120.  doi: 10.16098/j.issn.0529-1356.2017.01.021
    Abstract ( )   PDF (245KB) ( )  

    Chimeric antigen receptor modified T cells (CAR-T) has been an emerging immunotherapy modality for malignant tumor. The cytotoxicity efficiency of CAR-T cells to tumor is independent of the major histocompatibility complex (MHC). CAR-T cells can overcome the immunity suppression in the microenvironmet of the tumor site and break out the immunological tolerance of the host. Thus it has its own advantages in the immunotherapy for tumors. The construction of CAR-T cells and the selection of appropriate tumor target molecules are the two key challengs in CAR-T cell immunotherapy, this review will focus on these two issues.

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    Neurobiology
    Effects of resveratrol pretreatment on neurite growth of primary cortical neurons after oxygen-glucose deprivation/reperfusion injury in rats
    TANG Fan-ren YU Ping-ping WANG Li GUO Shuang YANG Qin
    2017, 48 (1):  1-6.  doi: 10.16098/j.issn.0529-1356.2017.01.001
    Abstract ( )   PDF (729KB) ( )  

    Objective To investigate the effects of resveratrol pretreatment on neurite growth of rat primary cortical neurons after oxygen-glucose deprivation/reperfusion (OGD/R) injury in vitro. Methods Primary cortical neurons were cultured under oxygen and glucose deprivation for 150 minutes and reoxygenation for 24 hours. The study had the normal, control and 5μmol /L resveratrol pretreatment groups. Neurons were identified with immunofluorescence. Cell viability was detected with cell counting kit-8(CCK-8) assay. Cell apoptosis was detected with TUNEL assay. Immunofluorescence and Western blotting measured the expressions of microtubule-associated protein 2(MAP-2)and growth associated protein 43(GAP-43), and the length and number of neurites were counted.
    Results Cells had high expression of neuronal specific marker MAP-2. Compared with the control group, resveratrol treatment significantly enhanced the neurons viability (0.551±0.009 vs 0.436±0.013,P<0.01), decreased the numbers of apoptosis (18.3%±1.3% vs 35.3%±1.9%,P<0.01), upregulated the expressions of MAP-2 (0.790±0.102 vs 0.462±0.063,P<0.01) and GAP-43 (0.768±0.084 vs 0.424±0.065,P<0.01) proteins, increased the length (89.510±6.939 vs 61.538±9.14,P<0.01) and numbers (6.347±1.002 vs 3.040±0.608,P<0.01) of neurites. Conclusion Resveratrol pretreatment can reduce injury and promote neurite growth of cultured neurons after OGD/R.

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    Construction of Wnt3a fusion protein vector and its effect on the proliferation and axon formation of neural precursor cells during the chick embryonic spinal cord development
    LI Qiu-ling YANG Ci-qing ZHANG Bi-chao ZHU Shao-yi LIN Jun-tang
    2017, 48 (1):  7-13.  doi: 10.16098/j.issn.0529-1356.2017.01.002
    Abstract ( )   PDF (1161KB) ( )  

    Objective To construct a eukaryotic vector of chicken-derived Wnt3a tagged with EGFP (pCAG-MCs-Wnt3a-EGFP) and investigate the influence to the proliferation and axonal formation of neural precursor cells when Wnt3a was overexpressed during the development of chick embryonic spinal cord. Methods Wnt3a gene was amplified from the total RNA obtained from chick embryonic spinal cord using molecular techniques, then connected with pCAG-MCs-EGFP to construct pCAG-MCs-Wnt3a-EGFP, which was identifled by digestion and genetic sequencing. At embryonic day (E) 2.5-3.0, pCAG-MCs0-Wnt3a-EGFP (experimental group) and pCAG-MCs-EGFP (control group) were transfected into the chick embryonic spinal cord using in vivo electroporation, respectively. Samples were collected at E4 (5 simples of each groups ) and then conducted frozen section. The immunofluorescent staining was performed to detect the expression of Wnt3a and proliferating cell nuclear actigen (PCNA) for analyzing the relationship between Wnt3a and cell proliferation, and observe the axonal formation of neural precursor according to the green fluorescence of Wnt3a protein. Results pCAG-MCs-Wnt3a-EGFP was obtained and its gene sequencing was identical with the Gene bank. Green fluorescence was observed at E4 after pCAG-MCs-Wnt3a-EGFP transformed to chick spinal cord. In transversal section of chick embryonic spinal cord, the results of immunofluorescent staining showed Wnt3a was successfully overexpressed. Meanwhile, the amount of neurons projecting axons was dramatically decreased (n=3, P<0.01), compared to the control group, concomitant with the significant elevation of PCNA level (n=3, P<0.01). Conclusion pCAG-MCs-Wnt3a-EGFP is successfully constructed and our study confirmed that Wnt3a plays a vital role in the proliferation and axonal formation of neural precursor cells in the developing chick spinal cord.

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    Intermittent starvation attenuating neurofilament phosphorylation and spatial learning and memory disorder induced by sodium nitrite in rats
    HU Zhi-hong YAN Jun-bao LI San-qiang HU Yong-mei
    2017, 48 (1):  14-18.  doi: 10.16098/j.issn.0529-1356.2017.01.003
    Abstract ( )   PDF (505KB) ( )  

    Objective To explore the effect of intermittent starvation on neurofilament phosphorylation and spatial learning and memory in rats induced by sodium nitrite.Methods Thirty-six rats were randomly divided into control, NaNO2 and stavation+NaNO2 groups, 12 rats in each group. NaNO2 group and stavation+NaNO2 group rats were served as drinking water containing sodium nitrite [100mg/(kg·d)]. Starvation+NaNO2 group rats were deprived of food for 2 days, and then re-feeded for 3 days. Sixty days later, the ability of spatial learning and memory of the rats was measured by Morris water maze. Phosphorylation level of neurofilament in the hippocampus was detected by immunohistochemistry and Western blotting. Results In comparison with the control rats, the NaNO2 rats showed significantly longer latency to find the hidden platform from 2 days to 7 days in Morris water maze (control group latency: 31.24±8.53,12.41±6.54,10.49±6.43,8.61±2.56,7.25±2.12,6.03±1.92; NaNO2 group latency: 53.34±5.28,35.15±10.29,23.52±9.50,14.49±8.70,16.87±8.77,12.31±7.12) (P<0.05,P<0.01), and elevated phosphorylation level of neurofilament, while stavation+NaNO2 group rats only showed longer latency on 2 days and 3 days(43.61±1.76,25.25±7.49)in Morris water maze. There was no obvious difference in phosphorylation level of neurofilament between the control and stavation+NaNO2 group. Conclusion Intermittent starvation attenuates neurofilament phosphorylation and spatial learning and memory disorder induced by sodium nitrite in rats.

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    Expression and alteration induced by MPTP / MPP+of mitochondrial transcription termination factors family
    ZHANG Lin-bing HAN Yan-yan QIU Shi XU Shan ZUO Ji LIU Wen
    2017, 48 (1):  19-24.  doi: 10.16098/j.issn.0529-1356.2017.01.004
    Abstract ( )   PDF (627KB) ( )  

    Objective To investigate the expression of mitochondrial transcription termination factors (MTERFs) in various tissues and cells, and the changes of MTERFs induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)/1-methyl-4-pheny lpyridine(MPP+ ). Methods Western blotting was used to measure the expression of the MTERFs in brain, liver, kidney of C57BL mice and in human SH-SY5Y, HCM, L-02 cells. MPTP/MPP+ was injected to imitate PD animal/cell models. The changes of MTERFs protein were detected through western blotting. Results The expression of MTERFs was higher in mice brain and SH-SY5Y cells. In PD mouse model induced by MPTP, MTERF3 reduced significantly. Conclusion The expression of MTERFs was higher in mouse brain and SH-SY5Y cells. After induced by MPTP/MPP+ , MTERF3 decreased obviously, which increased transcription level and might involve in a process of Parkinson's disease.

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    Effect of microRNA-181b on ischemic stroke  by regulating the heat shock protein A5 in mice
    PENG Zhi-feng
    2017, 48 (1):  25-29.  doi: 10.16098/j.issn.0529-1356.2017.01.005
    Abstract ( )   PDF(mobile) (419KB) ( 0 )  

    Objective To explore the role of microRNA-181b (miR-181b) in cerebral ischemic injury in vivoand its mechanism. Methods Using middle cerebral artery occlusion (MCAO) model to mimic ischemic injury in vivo, the heat shock protein A5(HSPA5)protein level was determined by using Western blotting. The extent of neural cell loss in ischemic cortex after MCAO was assessed by Nissl staining. Neurological score was performed to evaluate the degree of cerebral ischemic injury after MCAO. Results We found that miR-181b antagomir down-regulated miR-181b expression levels in cerebral ischemic cortex of mice after MCAO(P<0.05,n=3). MiR-181b antagomir improved neurological deficit of mice at 24 hours after transient MCAO (P<0.05,n=6). HSPA5 protein levels were significantly up-regulated in ischemic cortex of mice after MCAO,and miR-181b antagomir further up-regulated HSPA5(P<0.05,n=3).Consequently, miR-181b antagonists attenuated neural cell loss in ischemic cortex after MCAO(P<0.05,n=3). Conclusion MiR-181b plays an important role in ischemic injury of mice through regulating HSPA5 protein level.

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