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    Original article (Basic research)
    Immunological evaluation of subunit vaccine prepared by combining specific antigen Rv3117 of Mycobacterium tuberculosis with DDA/MPL adjuvant
    YE Juan, GAO Meng-zhe, ZHANG Shu-lin, et al
    2015, 35 (1):  6. 
    doi: 11.3969/j.issn.1674-8115.2015.01.001

    Abstract ( 737 )   PDF (529KB) ( 1135 )  

    Objective To construct a TB subunit vaccine (Rv3117/DDA/MPL) by combining specific antigen Rv3117 of Mycobacterium tuberculosis (M. tuberculosis) with the adjuvant of dimethyl dioctadecyl ammoniumbromide (DDA) /monophosphoryl lipid A (MPL) and to evaluate the immune responses after the bacillus calmetteguerin (BCG)-Prime in C57BL/6 mice. Methods The molecular cloning method was adopted to construct a recombinant plasmid pET32a-Rv3117 and the specific recombinant Rv3117 protein of M. tuberculosis expressed in Escherichia coli BL21(DE3) PLysS was obtained. Then the antigen Rv3117 was induced and purified. The endotoxin was removed by Triton X-114 phase separation method and the subunit vaccine was prepared by fully emulsifying M.tuberculosis specific antigen Rv3117 with DDA/MPL adjuvant. The C57BL/6 mice were randomly divided into the control group (without any treatment), PBS group (immunized with PBS), BCG group (only immunized with BCG), BCG+DDA/MPL group (immunized with DDA/MPL adjuvant twice after being immunized with BCG for 2 weeks), and BCG+Rv3117/DDA/MPL group (immunized with subunit vaccine Rv3117 twice after being immunized with BCG for 2 weeks) (n=5/group). The humoral immunity and cellular immunity were evaluated by the enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunospot Assay (ELISPOT), respectively. Results The specific antigen Rv3117 of M. tuberculosis was successfully coloned and purified. The results of ELSPOT and ELISA indicated that level of interferon γ (IFN-γ) produced by T lymphocytes of mice of the BCG+Rv3117/DDA/MPL group was significantly higher than that of the control group, PBS group, BCG group, and BCG+DDA/MPL group (P<0.05) after being stimulated by the purified protein derivative (PPD) of tuberculin (10.0 μg/mL). The levels of tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), and IL-12p70 of culture supernatants of the BCG+DDA/MPL group were significantly higher than those of the control group and PBS group (P<0.05) and the total level of IgG and IgG1 was also higher. The levels of above indexes of the BCG+Rv3117/DDA/MPL group were significantly better than those of the BCG+DDA/MPL group (P<0.05). Conclusion The TB subunit vaccine Rv3117/DDA/MPL can strengthen the humoral immunity and cellular immunity of BCG in C57BL/6 mice. The specific antigen Rv3117 of M. tuberculosis is expected to be a novel candidate antigen for designing the TB vaccine and diagnosing the TB.

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    Effects of Rhizoma Atractylodis before and after being fried with bran on immune system and gastrointestinal hormones of rats with experimental spleen deficiency
    LIU Fen, LIU Yan-ju, TIAN Chun-man
    2015, 35 (1):  8. 
    doi: 11.3969/j.issn.1674-8115.2015.01.002

    Abstract ( 435 )   PDF (421KB) ( 1035 )  

    Objective To explore the effects of Rhizoma Atractylodis before and after being fried with bran on the gastrointestinal motility and immune function of rats with experimental spleen deficiency syndrome and to confirm if Rhizoma Atractylodis fried with bran can strengthen the effect of Rhizoma Atractylodes on interventing the spleen deficiency syndrome and possible mechanisms. Methods Rats were fed with Xiaochengqi Decoction combined with irregular hunger method to establish the spleen deficiency syndrome model. After the model was successfully copied, rats were randomly divided into the normal group, model group, Rhizome Atractylodes group, Rhizome Atractylodes fried with bran group, and domperidone group. After 10 d of treatment, the residual rate of stomach and small intestinal propulsion ratio of the rats were detected by the charcoal gavage method. The serum gastrin (GAS), motilin (MTL), and vasoactive intestinal peptide (VIP) levels were detected by the ELISA method. The serum interleukin (IL)-1, -2, -6 and tumor necrosis factor (TNF)-α levels were detected by the radioimmunoassaymethod. The indexes of spleen and thymus and the proliferation rate of spleen lymphocytes (T, B) were measured. Results Compared to the normal group, the gastrointestinal dynamics, hormone expressions, and immune function of the model group were significantly abnormal. Compared to the model group, residual rates of stomach of the Rhizome Atractylodes group, Rhizome Atractylodes fried with bran group, and domperidone group significantly decreased; small intestinal propulsion ratios significantly increased; serum GAS, MTL, and VIP levels increased with different degree; serum IL-1, -2, -6 and TNF-α levels also increased with different degree; and indexes of spleen and thymus and proliferation rates of spleen lymphocytes (T, B) increased with different degree and the differences were statistically significant (P<0.05 or P<0.01). Compared to the Rhizome Atractylodes group, the effects of Rhizome Atractylodes fried with bran were more significant and the differences of two groups were statistically significant (P<0.05 or P<0.01). Conclusion Rhizome Atractylodes extract can improve the gastrointestinal motility and regulate gastrointestinal hormone secretion and immune function of experimental spleen deficiency rats. The effects of Rhizome Atractylodes fried with bran are better than those of Atractylodes rhizome.

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    Inhibition of green tea polyphenol on angiogenesis of chick chorioallantoic membrance
    LIU Chang, SHEN Xiu-hua, TANG Wen-jing, et al
    2015, 35 (1):  13. 
    doi: 11.3969/j.issn.1674-8115.2015.01.003

    Abstract ( 511 )   PDF (493KB) ( 953 )  
    Objective To observe the effect of green tea polyphenol extract, (-)-epigallocatechin-3-gallate (EGCG), on anti-angiogenesis. Methods Fresh chicken eggs with normal chicken embryos were randomly divided into the blank control group, PBS group, low dose EGCG group, medium dose EGCG group, and high dose EGCG group after being incubated for 5 d and each group consisted of 7 eggs. Then, 10 μL sample was injected through the air chamber of eggs (EGCG solutions of 100, 1 000, and 10 000 μg/mL were used for the low dose, medium dose, and high dose EGCG groups, respectively). Chick chorioallantoic membrane (CAM) was solidified and cut out after 2 d. The angiogenesis of CAM of each group was photographed recorded and the vascular intersection number (VN) was manually counted. The vessel angiogenesis area (VA) was scanned and analyzed and the ratio of VA and CAM (VA/CAM) was calculated. Results Blood vessels of CAM of the blank control group and PBS group were abundant and thick. With the increase of EGCG concentration, blood vessels were gradually sparse and scattered. VNs of 1 cm×1 cm area of the PBS group, low dose EGCG group, medium dose EGCG group, and high dose EGCG group were 9.5±1.29, 9.8±1.30, 8.2±1.30, and 6.5±0.58, respectively. The differences of the high dose EGCG group and other groups were statistically significant (P<0.05). VA/CAM values of the PBS group, low dose EGCG group, medium dose EGCG group, and high dose EGCG group were 0.24±0.04, 0.18±0.01, 0.14±0.02, and 0.08±0.01, respectively. The differences among groups were statistically significant (P<0.01). Conclusion Green tea polyphenol can inhibit the angiogenesis of CAM with dose-dependent effect.
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    Effects of ursolic acid on activation of NADPH oxidase and downstream signaling pathways of hepatic stellate cells induced by angiotensinⅡ
    CHEN Tao, HE Wen-hua, ZHU Xuan, et al
    2015, 35 (1):  17. 
    doi: 11.3969/j.issn.1674-8115.2015.01.004

    Abstract ( 448 )   PDF (467KB) ( 887 )  
    Objective To analyze the effects of ursolic acid (UA) on the activation of NADPH oxidase (NOX) and the downstream signaling pathways of PI3K/Akt and P38MAPK of hepatic stellate cells (HSC) induced by angiotensinⅡ(AngⅡ). Methods Culture-activated HSC-T6 cells were divided into the control group (received no medicines), AngⅡ group (received AngⅡ of 1 μmol/L), and 4 intervention groups, which were pretreated with UA of 50 μmol/L, NOX inhibitor DPI of 20 μmol/L, PI3K/Akt inhibitor LY294002 of 10 μmol/L, and P38MAPK inhibitor SB203580 of 10 μmol/L for 30 min and then treated with AngⅡ for different periods of time. Expressions of NOX subunit p47phox, total PI3K, phosphorylated p-Akt, and p-P38MAPK of the membrane were detected by the Western blotting. Collagen Ⅰ mRNA expressions induced by AngⅡ were detected by the RT-PCR and the proliferation rate of HSC-T6 was measured by the CCK-8. Results After being treated with AngⅡ for 15 min, the expression of p47phox of membrane was significantly higher than that of the control group (P<0.05). After being intervened by DPI and UA, the expression of p47phox of membrane was significantly lower than that of the AngⅡ group (P<0.05). After being treated with AngⅡ for 30 min, the expressions of PI3K and p-Akt were significantly higher than those of the control group (P<0.05). After being intervened by UA, DPI, and LY294002, the expressions of PI3K and p-Akt were lower than those of the AngⅡ group (P<0.05). After being treated with AngⅡ for 30 min, the expression of p-P38MAPK was significantly higher than that of the control group (P<0.05).After being intervened by SB203580, UA, and DPI, the expression of p-P38MAPK was significantly lower than that of the AngⅡ group (P<0.05). After being treated with AngⅡ for 12 h, the mRNA expression of collagenⅠ was significantly higher than that of the control group (P<0.05). After being intervened by SB203580, LY294002, UA, and DPI, the mRNA expression of collagenⅠ was significantly lower than that of the AngⅡ group (P<0.05). After being treated with AngⅡ for 12, 24, and 48 h, the proliferation rate of HSC-T6 significantly increased. After being intervened by LY294002, SB203580, UA, and DPI, the proliferation rate HSC-T6 was lower than that of the AngⅡ group (P<0.05). Conclusion UA can block the signal transduction of AngⅡ in HSC by inhibiting the activation of NOX subunit p47phox, therefore inhibit the proliferation of HSC and the mRNA expression of CollagenⅠ.
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    Protective effect of fibroblast growth factor 21 on cardiomyocyte hypertrophy induced by high glucose
    MENG Zhe-ying, WANG Yu, LIN Yan-duan, et al
    2015, 35 (1):  23. 
    doi: 11.3969/j.issn.1674-8115.2015.01.005

    Abstract ( 605 )   PDF (529KB) ( 939 )  
    Objective To observe the expression of fibroblast growth factor 21 (FGF21) for inducing the primary myocardial cells by high glucose and to explore the protection effect of endogenous FGF21 on the cardiomyocyte hypertrophy induced by high glucose. Methods Cardiomyocytes of neonatal rat C57BL/6J were primarily cultured and expressions of FGF21 were detected for different concentrations of glucose (5, 25, 33, and 45 mmol/L) and different exposure time (24 and 48 h). The small interfering RNA (siRNA) for silencing the FGF21 gene was designed and synthesized. Morphological changes of cardiomyocytes were observed and expressions of hypertrophy phenotype ANP and β-MHC mRNA of cardiomyocytes were detected by the Real-time PCR. Results The FGF21 level increased with the concentration of glucose. The FGF21 level of high glucose groups (33 and 45 mmol/L) was higher than that of the control group at the same time point (P<0.05). With the increase of exposure time, the FGF21 level increased first and then decreased. The FGF21 level at 24 h increased and was higher than that at 48 h (P<0.05). The expression of FGF21 mRNA of high glucose groups (33 and 45 mmol/L) was higher than that of the control group. With the increase of exposure time, the expression of FGF21 mRNA increased first and then decreased (P<0.05). After being treated by FGF21-siRNA, the cardiomyocyte hypertrophy induced by high glucose was more significant and expressions of ANP and β-MHC increased (P<0.05). Conclusion The expression of endogenous FGF21 depends on the concentration of glucose to a certain extent under high glucose condition. FGF21 may involve in the process of cardiomyocyte hypertrophy induced by high glucose and protect cardiomyocytes against the hypertrophy.
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    Evaluation of physicochemical and biological properties of modified keratin/silk fibroin compound biomaterial in vitro
    FENG Chao, LI Zhe, Lü Xiang-guo, et al
    2015, 35 (1):  29. 
    doi: 11.3969/j.issn.1674-8115.2015.01.006

    Abstract ( 637 )   PDF (1030KB) ( 1127 )  

    Objective To explore the feasibility of modified technique for the keratin/silk fibroin compound biomaterial, identify physicochemical properties, and evaluate biological properties. Methods The keratin and silk fibroin were extracted from human hair and worm silk, respectively. The SDS-PAGE was adopted to determine the properties of those two proteins. Two proteins were mixed at different concentrations and different concentrations of gelatin were added. The tensile mechanical tests were used to detect the keratin/silk fibroin compound biomaterial and then the optimal combination of protein concentration and gelatine concentration was screened. Variations of the spatial structure of proteins were analyzed by the Fourier transform infrared spectroscopy (FTIR). The compound biomaterial was embedded beneath the rabbit skin to evaluate its biocompatibility and the cytotoxicity was detected by the MTT assay. Results The concentration and purity of keratin and silk fibroin met the requirements of subsequent tests after extraction and purification. Mechanical tests showed that best mechanical properties were achieved when the concentration of gelatin was 2% and the ratio of keratin and silk fibroin was 60∶40. Results of the FTIR indicated that the spatial structure of each protein of the compound biomaterial did not vary significantly. Tests of embedding the compound biomaterial beneath the rabbit skin showed that the biocompatibility and degradation rate of the compound biomaterial were ideal. Results of the MTT indicated that the compound biomaterial has no significant cytotoxicity. Conclusion The physicochemical properties of reported keratin/silk fibroin compound biomaterial can be improved by proper ratio of keratin and silk fibroin and proper concentration of gelatine, which is more suitable for subsequent fundamental researches and clinical applications of repair and reconstruction of tissue engineering.

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    Effects of adenanthin to cell death and expression of telomerase through peroxiredoxin Ⅰ
    CUI Jia-yi, CAO Yang, LEI Hu, et al
    2015, 35 (1):  36. 
    doi: 11.3969/j.issn.1674-8115.2015.01.007

    Abstract ( 717 )   PDF (728KB) ( 1006 )  

    Objective To investigate the effects of adenanthin on telomerase of leukemia NB4-LR1 cells and relevant mechanisms. Methods NB4-LR1 cells were treated with different concentrations of adenanthin (0, 2, 4, and 6 μmol/L) for different periods of time (0, 6, 12, and 24 h). The expression of human telomerase reverse transcriptase (hTERT) protein was detected by Western blotting, and the expression of hTERT mRNA was detected by Real-time PCR. The RNA interference method was adopted to decrease the expression of peroxiredoxin Ⅰ (Prdx Ⅰ) in NB4-LR1 cells. Then the expression of hTERT protein was detected by Western blotting. hTERT was over-expressed in NB4-LR1 cells and the NB4-LR1-hTERT-GFP and NB4-LR1-GFP cells were treated with different concentrations of adenanthin (2 and 4 μmol/L). The variations of proliferation and viability of cells were detected by trypan blue exclusion assay. Results The results of Western blotting showed that adenanthin inhibited the expression of hTERT in a dosetime dependent manner. The expression of hTERT decreased after Prdx I was knocked down. Over-expression of hTERT in NB4-LR1 cells partially inhibited the toxic effect of adenanthin on cells, but could not inhibit cell differentiation induced by adenanthin. Conclusion Adenanthin inhibits the expression of telomerase through Prdx Ⅰ and contributes to the death of cells.

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    Effects of methylation of RFC1 gene of U937 cell line on cell cycle and apoptosis
    ZHANG Na, JIANG Hui, LI Hong, et al
    2015, 35 (1):  41. 
    doi: 11.3969/j.issn.1674-8115.2015.01.008

    Abstract ( 561 )   PDF (693KB) ( 1040 )  

    Objective To investigate the methylation, expression, and transcription of the promoter of reduced folate carrier 1 (RFC1) gene of U937 cell line and to observe the effects of variations of the methylation level on the cell apoptosis and cycle induced by methotrexate (MTX). Methods The quantitative real-time methylation-specific PCR and real-time quantitative PCR were adopted to detect the methylation status and mRNA expression of RFC1 gene promoter of U937 cell line after being incubated by 5-azacytidine. The effects of variations of RFC1 gene expression on the apoptosis and cell cycle of U937 cells induced by MTX were verified by the flow cytometry. Results The un-methylation rate of RFC1 gene promoter of U937 cells was (8.09±0.87)% which was in high methylation status and increased to (9.44±2.01)%, (13.51±2.19)%, and (15.76±1.65)% after being demethylated by 5-azacytidine of 1, 2, and 4 μmol/L for 72 h, respectively. Compared with the control group, the un-methylation rate of RFC1 promoter significantly increased (P=0.005, P=0.001) and the mRNA expression of RFC1 promoter increased (P=0.003, P=0.000) after being treated by 5-azacytidine of 2 and 4 μmol/L. And 5-azacytidine of 2 μmol/L inhibited the S phase and increased G1 phase of cell cycle. MTX of 5 nmol/L induced the apoptosis and inhibited the S phase of cell cycle. The combination of MTX and 5-azacytidine significantly increased the apoptosis and inhibited S phase of cell cycle. Conclusion The methylation level of RFC1 gene promoter of U937 cell line is abnormally high. And 5-azacytidine can demethylate and increase the mRNA expression of RFC1 gene promoter, therefore enhance the apoptosis effect of MTX and specifically inhibit the S phase of cell cycle.

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    Effects of andrographolide on proliferation and apoptosis of NB4 cells in vitro
    TANG Yu-hong, WANG Yu-li, LU Jing-feng, et al
    2015, 35 (1):  47. 
    doi: 11.3969/j.issn.1674-8115.2015.01.009

    Abstract ( 510 )   PDF (411KB) ( 1009 )  

    Objective To investigate the effects of andrographolide (AD) on the cell proliferation and apoptosis of human acute promyelocytic leukemia-NB4 cells in vitro. Methods NB4 cells were treated with AD of different concentrations for 48 h. The cell proliferation was detected by the MTT assay and the early apoptosis rate was determined by the AnnexinV/PI double staining. The subdiploid peak and cell cycle were analyzed by the DNA-PI staining. The expressions of apoptosis related proteins were detected by the Western blotting. Results AD of 5-25 μmol/L significantly inhibited the proliferation of NB4 cells. The half inhibit concentration (IC50) of AD at 48 h was (1.98±0.22) μmol/L. With the increasing of AD concentration, the early apoptotic rate, subdiploid peak proportion, and apoptosis proportion elevated significantly (P<0.05). The results of Western blotting showed that the expressions of apoptosis related proteins were significantly activated. Conclusion AD can effectively inhibit the proliferation of NB4 cells, induce cell apoptosis, and arrest the cell cycle.

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    Study on inhibition of fibrosis of C2C12 cells in vitro by miRNA138
    XUE Ming-feng, GONG Sui-liang, Dai Jia-ping, et al
    2015, 35 (1):  51. 
    doi: 11.3969/j.issn.1674-8115.2015.01.010

    Abstract ( 581 )   PDF (719KB) ( 1082 )  

    Objective To explore the establishment of a fibrosis model in vitro by inducing rat myoblast C2C12 by transforming growth factor-β (TGF-β), observe the effects of variations of the miRNA138 level on cell fibrosis indexes, and analyze whether the effects of miRNA138 on cell fibrosis was achieved by its target gene Smad4. Methods The miRNA138-mimics, miRNA138-inhibitor, and miRNA138-NC were designed, synthesized and transfected into C2C12 cells by lipofectminae. Transfected cells were then induced by TGF-β1 of 10 ng/mL for 48 h. The expression of miRNA138 was detected by Real-time PCR. The variations of expressions of fibrosis related proteins Smad4, vimentin, α-SMA, and collagen I were detected by the Western blotting to verify whether the fibrosis model had been successfully established and analyze the effects of variations of the miRNA 138 expression on cell fibrosis indexes. The effects of variations of the miRNA 138 expression on the proliferation activity of C2C12 during the course of fibrosis were measured by the CCK-8. Bioinformatic software was used to analyze the theoretical binding site of miRNA138 and Smad4, which was verified by the luciferase reporter gene test. The Smad4 gene expression vector was constructed and co-transfected into C2C12 cells with miRNA-mimics. Transfected cells were induced by TGF-β1 for fibrosis. The variations of fibrosis related proteins were detected by the Western blotting and the effects of expression of exogenous Smad4 on the inhibition of cell fibrosis by miRNA138 were analyzed. Results The expression of miRNA138 significantly increased after C2C12 cells were transfected by miRNA138-mimics and decreased after C2C12 cells were transfected by miRNA138-inhibitor. The expression of miRNA138 of the control group had no significant changes. Results of protein detection showed that expressions of Smad4, vimentin, α-SMA, and collagen I were significantly increased and the cell proliferation activity in logarithmic phase was enhanced after C2C12 cells were induced by TGF-β1 for 48 h, which indicated that the cell fibrosis model was successfully established. The over-expression of miRNA138 significantly inhibited the fibrosis of C2C12 cells. The data of luciferase activity detections showed that the predicted binding site of miRNA138 and Smad4 existed. High expression of exogenous Smad4 in C2C12 cells significantly impaired the inhibition of cell fibrosis by miRNA138-mimics. Conclusion MiRNA138 can inhibit the fibrosis of C2C12 cells induced by TGF-β1 via suppressing the expression of its target protein Smad4.

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    Original article (Clinical research)
    Analysis of gene expression and polymorphism of allele HLA-A*02 of Han patients with nasopharyngeal carcinoma in Xinjiang
    LIU Kai, TAN Yao, LI Ya-wei, et al
    2015, 35 (1):  59. 
    doi: 11.3969/j.issn.1674-8115.2015.01.011

    Abstract ( 602 )   PDF (247KB) ( 848 )  

    Objective To explore the relationship between the polymorphism of allele HLA-A*02 and the incidence of nasopharyngeal carcinoma (NPC) of Han patients in Xinjiang. Methods Polymerase chain reaction/sequence-specific primers (PCR-SSP) method was adopted to analyze HLA-A alleles of 63 Han patients with NPC in Xinjiang and 51 health Han volunteers. The differences of allele HLA-A*02 and its subtypes of two groups were compared. Results The frequency of allele HLAA*02 in NPC patients was 39.7% (51/126) and the frequency of allele HLA-A*02 in healthy people was 16.7% (17/102). The difference was statistically significant (P=0.002). Seven subtypes of allele HLA-A*02 were detected in NPC patients and 4 subtypes of allele HLA-A*02 were detected in healthy people. The difference was not statistically significant (P>0.05). The differences of constituent ratios of subtypes were not statistically significant (P>0.05). Conclusion The allele HLA-A*02 is relevant to the incidence of NPC of Han patients in Xinjiang. The distribution of subtypes of allele HLA-A*02 of NPC patients in Xinjiang have their own features.

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    Effects of residual renal function before hemodialysis on long-term prognosis of patients undergoing hemodialysis
    ZHANG Yu-mei, BIAN Zhi-xiang, CHEN Pei-hua, et al
    2015, 35 (1):  63. 
    doi: 11.3969/j.issn.1674-8115.2015.01.012

    Abstract ( 745 )   PDF (445KB) ( 1021 )  

    Objective To evaluate the relationship between the residual renal function before hemodialysis and the long-term prognosis of patients who undergo hemodialysis. Methods Data of adult patients with the end-stage renal disease (ESRD) who underwent the maintenance hemodialysis between January, 2005 and September, 2013 was collected. Patients were followed up till March 31, 2014. According to the estimated glomerular filtration rate (eGFR) before hemodialysis, patients were divided into four groups and their eGFRs were ≥10.5, 8.1-10.4, 6-8, and <6 mL·min-1·(1.73 m2)-1, respectively. The simplified equation of modification of diet in renal disease (MDRD) was adopted to evaluate the eGFR. The endpoint event was all-cause mortality. Results The median eGFR of 294 patients before hemodialysis was 5.43 (2.27-13.92) mL·min-1·(1.73 m2)-1. A total of 65 patients died during the period of follow-up and the top three causes of death were cerebrovascular accidents, infections, and cardiovascular diseases. With the decrease of eGFR after hemodialysis, blood urea nitrogen, serum creatinine, serum uric acid, phosphorus, calcium and phosphate product, serum ferritin, proportion of hyperuricacidemia complication, and proportion of Charlson combidity indexes (CCI) between 0-2 gradually increased and proportion of diabetes, hematocrit, and proportion of CCI≥5 gradually decreased. The multivariate Cox regression analysis indicated that the serum creatinine was a protective factor of long term prognosis of patients who underwent the hemodialysis and CCI, combined cerebrovascluar diseases, combined chronic obstructive pulmonary disease, emergent hemodialysis due to other reasons, and triglyceride were risk factors. The stratified analysis was performed for the emergency hemodialysis group, selecting hemodialysis group, albumin <35 g/L group, albumin ≥35 g/L group, age <65 years group, and age ≥65 years group. The results showed that the long-term prognosis of patients with high eGFRs before hemodialysis was not better. Conclusion Although the incidences of anemia, hyperuricacidemia, and calcium and phosphorus metabolism disorder of patients with low eGFRs before hemodialysis are more than those of patients with high eGFRs, the long-term prognosis of patients with high eGFRs before hemodialysis is not better. There is no significant relationship between the eGFR before hemodialysis and the long term prognosis.

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    Correlative factors and therapeutic effect of one treatment or multi-treatments to Graves disease by 131I
    MA Yu-bo, PAN Yi-fan, GU Ai-chun, et al
    2015, 35 (1):  69. 
    doi: 11.3969/j.issn.1674-8115.2015.01.013

    Abstract ( 521 )   PDF (256KB) ( 949 )  

    Objective To retrospectively analyze correlative factors and therapeutic effect for the treatment to Graves disease by 131I and to identify critical influencing factors of one treatment and multi-treatments. Methods A total of 455 outpatients with Graves disease underwent the 131I treatment. The dose of 131I was determined based on the dose formula and clinical experience. For unrecovered or recurrent patients, the same treatment was repeated after 3 months or more until they were cured. Patients were followed up for more than 1 year after the treatment. The differences of correlative factors and therapeutic effect of patients cured by 1 treatment or multi-treatments were retrospectively analyzed and critical influencing factors and their clinical values were identified. Results There were 367 patients (80.7%) and 88 patients(19.3%) who were cured by one treatment (one treatment group) and multi-treatments (multi-treatment group), respectively. The differences of age, gender, 24h 131I uptake rate, withdrawal time of antithyroid drugs (ATD), thyroid hormone and autoantibody levels before treatment, and thyroid weight of two groups were not statistically significant (P>0.05). The activity values of the first 131I treatment of one treatment group and multi-treatment group were (314.5±136.9) MBq and (292.3±96.2) MBq, respectively (t=2.590 7, P<0.01). The ratios of 131I dose of the first treatment and thyroid weight of two groups were (3.8±1.2) MBq/g and (3.5±1.0) MBq/g (t=2.105 4, P<0.05). The total activity values of two groups were (314.5±136.9) MBq and (706.7±399.6) MBq (t=-13.283 0, P<0.01). There were 186 patients (50.7%) of one treatment group and 48 patients (54.5%) of multi-treatment group suffered from the permanent hypothyroidism (χ2=0.283 7, P>0.05). Conclusion For the Graves disease, one 131I treatment is better than multi 131I treatments. The critical influencing factor is that the activity of the first 131I treatment should be sufficient. Average MBq/g ratio ≥4.4 is recommended.

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    Therapeutic effect of imatinib on children with chronic myelogenous leukemia
    LI Shan-shan, JIANG Hui
    2015, 35 (1):  73. 
    doi: 11.3969/j.issn.1674-8115.2015.01.014

    Abstract ( 551 )   PDF (548KB) ( 1099 )  
    Objective To retrospectively analyze the therapeutic and adverse effects of imatinib (IM) on children with chronic myelogenous leukemia (CML). Methods The clinical data of 5 children who were diagnosed as CML combined with positive Philadelphia chromosome according to the MICM standard and treated with IM from Jan. 2012 to Jan. 2013 were collected. Among 5 cases, there were 2 males  and 3 females. The median age was 9 (6-11) years old. IM of 340 mg/(m2·d) was orally administrated and the median follow-up time was 25 (15-30) months. The clinical manifestations, variations of indexes of peripheral hemogram, bone marrow cytology, and molecular biology, as well as growth and development and bone metabolism were observed regularly. Results After being treated with IM, all patients attained complete hematological response (CHR) and complete cytogenetic response (CCyR) in 3 and 12 months, respectively. The serum osteocalcin level of 1 patient was one time higher than the upper limit of normal level after being treated with IM for 25 months. Indexes relevant to growth and development of other patients showed no significant abnormalities. Conclusion The therapeutic effect of IM on children with CML is significant and safe and the adverse effects should be observed through the long-term follow-up.
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    Clinical observation of interventional therapy of thyroid tumor by 32Phosphorus-radiocolloid
    WANG Jian, LOU Cen
    2015, 35 (1):  78. 
    doi: 11.3969/j.issn.1674-8115.2015.01.015

    Abstract ( 625 )   PDF (246KB) ( 1005 )  
    Objective To observe the effects of interventional therapy on the thyroid tumor by intratumoral injection of 32Phosphorus-radiocolloid (32P-colloidal) under ultrasonic guidance. Methods Patients with thyroid tumors who underwent intratumoral injection of 32P-colloidal (32P-colloidal group, n=30) and pure ethanol (control group, n=30) under ultrasound guidance were selected. The therapeutic effect and adverse reactions were recorded by follow-up after being treated for 2, 4, 6, and 8 months. Results Regular follow-up showed that the therapeutic effect of 32P-colloidal group was correlated with the tumor size after being treated for 6 and 8 months (P=0.042 and 0.019, respectively). The therapeutic effect of 32P-colloidal group was significantly better than that of the control group after being treated for 8 months (P=0.011). The main adverse reactions of 32P-colloidal group were mild pain at tumors sites (33.00%), local skin erubescence (16.67%), and radiodermatitis (6.70%). Conclusion The interventional therapy of thyroid tumor by intratumoral injection of 32P-colloidal under ultrasonic guidance is convenient and effective with less adverse reactions. The therapeutic effect is correlated with the tumor size.
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    Analysis of co-morbidities of chronic obstructive pulmonary disease for patients in Yangpu District of Shanghai
    CHENG Yi, HAN Xue, LUO Yong, et al
    2015, 35 (1):  82. 
    doi: 11.3969/j.issn.1674-8115.2015.01.017

    Abstract ( 616 )   PDF (323KB) ( 945 )  
    Objective To analyze the features of co-morbidities of chronic obstructive pulmonary disease (COPD) for residents in the Yangpu District of Shanghai. Methods According to the data provided by the Center for Disease Control and Prevention of the Yangpu District of Shanghai, patients whose death ages in medical certificates of death from 2003 to 2011 were ≥ 40 years and one of death causes was COPD were selected. The immediate cause of death (Part I of death causes of the medical certificate) and contributing cause (Part II of death causes of the medical certificate) were collectively designated as non-underlying causes of death. Results From 2003 to 2011, 63 161 patients whose ages were ≥ 40 years were died in the Yangpu District of Shanghai. A total of 8 838 patients died of COPD (underlying cause for 5 678 patients) and the average age was (81.3±7.6) years. Among them, there were 5 555 male patients and 2 783 female patients. When COPD was considered as the non-underlying causes of death, the common underlying causes of death were ischemic heart diseases (33.4%), cerebrovascular diseases (14.8%), lung cancer (14.4%), and diabetes (3.6%). With the increase of age, the proportion of cancer gradually decreased and the proportion of cardiovascular diseases gradually increased (P<0.05), while the proportion of cerebrovascular diseases did not change significantly (P>0.05). When COPD was considered as the underlying cause, the common non-underlying causes of death were pulmonary heart diseases (25.3%), ischemic heart diseases (23.2%), cerebrovascular diseases (13.9%), diabetes (8.8%), and cancer (5.5%). Conclusion Cardiovascular diseases, cancer, cerebrovascular diseases, and diabetes were common co-morbidities of COPD patients in Yangpu District of Shanghai.
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    Analysis of prognosis of surgical treatment to patients with Hirschsprung disease
    LI Long-zhi, XU Wei-jue, Lü Zhi-bao, et al
    2015, 35 (1):  86. 
    doi: 11.3969/j.issn.1674-8115.2015.01.017

    Abstract ( 679 )   PDF (354KB) ( 1092 )  
    Objective To analyze the effects of two surgical procedures on the postoperative complications and defecation function of patients with different pathologic types of Hirschsprung disease (HD). Methods The early postoperative complications of 101 children with HD who underwent radical procedures were retrospectively analyzed. The defecation function (Krechenbeck scores) and stooling frequency of 87 followed up cases were evaluated 6, 12, and 24 months after radical procedures. The relationships of two surgical procedures, different pathologic types, postoperative complications, and defecation function were assessed. Results The rate of early postoperative constipation of the Soave procedure (11.76%) was significantly higher than that of the Ikedasoper procedure (2.78%), while the rate of early pelvic inflammation complication of the Ikedasoper procedure (8.33%) was significantly higher than that of the Soave procedure (0%). The differences were statistically significant (P<0.05). The rate of postoperative complications increased with the severity of pathologic types. The defecation function improved with time. The differences of defecation function of different groups were not statistically significant 24 months after surgery (P>0.05). Conclusion Selection of the most appropriate procedure and management of postoperative diet and defecation are key points for the treatment of HD.
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    Fractionated stereotactic radiotherapy for treatment of locally recurrent nasopharyngeal carcinoma
    TANG Jian-min, HOU Yan-li, DAI Li-yan, et al
    2015, 35 (1):  91. 
    doi: 11.3969/j.issn.1674-8115.2015.01.018

    Abstract ( 446 )   PDF (403KB) ( 904 )  
    Objective To evaluate the therapeutic effect and adverse reactions of fractionated stereotactic radiotherapy (FSRT) for the treatment of locally recurrent nasopharyngeal carcinoma after radiotherapy by the gamma ray stereotactic radiotherapy. Methods A total of 30 patients with recurrent nasopharyngeal carcinoma were divided into two groups (A group and B group) and treated by FSRT. The prescription dose at 50% isodose line was 2 Gy each time and each patient consecutively underwent 5 treatments per week. The mean doses of A group (n=8) and B group (n=22) were 15 Gy and 40 Gy, respectively. Results The complete response (CR) and partial response (PR) rates of A group were 87.5% (7/8) and 12.5% (1/8) after being treated by FSRT for 3 months and the CR and PR rates of B group were 77.3% (17/22) and 22.7% (5/22). The follow-up time was 10-70 months and the median follow-up time was 26 months. The Karnofsky performance status scores for 95.2%(20/21) of the surviving patients were over 90. The one-year survival rate and local control rate of A group were 77.78% and 77.78% and two-year survival rate and local control rate were 51.85% and 77.78%. The one year survival rate and local control rate of B group were 90.23% and 90.23% and two-year survival rate and local control rate were 82.71% and 74.01%. Conclusion The local control effect of FSRT on locally recurrent nasopharyngeal carcinoma after
    radiotherapy is good and it is worth further clinical studies.
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    Original article (Public health)
    Qualitative study on motivators of medical staff
    WANG Meng, LI Guo-hong, JIN Chun-lin, et al
    2015, 35 (1):  97. 
    doi: 11.3969/j.issn.1674-8115.2015.01.019

    Abstract ( 553 )   PDF (550KB) ( 1008 )  
    Objective To provide suggestions for boosting the motivation of medical staff by understanding their motivators. Methods Semi-structured interviews on the working status of 89 medical staff from 6 cities in the eastern, central, and western region of China were conducted, including sources of pressure of current works, major demands, and understanding of relevant policies, etc. The data were analyzed by the Nvivo 10.0 software. Results Medical staff concerned more about “salary” and
    “society respect”. The demands of medical staff of different regions were different. Medical staff of central regions concerned more about “salary”; medical staff of eastern regions concerned more about “scientific research and education” and “work pressure”; while demands of medical staff of western regions were more balanced. Conclusion Motivating measures of medical institutions should focus on the regionalization and customization. Different salary models may be adopted for different positions and work features. The modern personnel management system of hospital should be established and medical liability insurance system should be promoted.
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    Relationships among personality characteristics, interpersonal trust, and subjective well-being of medical students
    YANG Xiu-mu, SHEN Zheng-fu, QI Yu-long, et al
    2015, 35 (1):  102. 
    doi: 11.3969/j.issn.1674-8115.2015.01.020

    Abstract ( 568 )   PDF (372KB) ( 930 )  

    Objective To investigate the relationship between personality characteristics and subjective well-being of medical students and the mediating effect of interpersonal trust on the relationship. Methods A total of 1 519 medical students were surveyed by the Eysenck Personality Characteristics Scale,  Interpersonal Trust Scale, and the Subjective Well-being Scale. Results The difference of scores of subjective well-being of male and female students was not statistically significant (t=-0.24, P>0.05). The scores of interpersonal trust of male students were significantly lower than those of female students (t=-2.68, P<0.01). Comparisons of different grades showed that the scores of subjective well-being of freshmen were significantly higher than those of sophomores and juniors (F=38.36, P<0.001). The scores of interpersonal trust of sophomores were significantly higher than those of freshmen and juniors (F=8.07, P<0.001). The interpersonal trust negatively correlated with psychoticism, extraversion, and subjective well-being (r=-0.089, r=-0.144, r=-0.122; P<0.05 or P<0.001) and positively correlated with neuroticism (r=0.163, P<0.001). The subjective well-being negatively correlated with psychoticism and neuroticism (r=-0.127, r=-0.348; P<0.001) and positively correlated with extraversion (r=0.171, P<0.001). The psychoticism and neuroticism negatively predicted the subjective well-being; the extroversion significantly and positively predicted the subjective well-being; the psychoticism and extraversion negatively predicted the interpersonal trust; the neuroticism positively predicted the interpersonal trust; and the interpersonal trust significantly and negatively predicted the subjective well-being (F=76.231, 21.253, 58.565 and R2=0.131, 0.041, 0.134, respectively). Conclusion The interpersonal trust partially mediates the relationship of personality characteristics and subjective well-being of medical students.

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    Spatial analysis of birth defects from 1996 to 2012 in China based on geographic information system
    WEN Xiao-yan, PENG Bin, HU Shan, et al
    2015, 35 (1):  107. 
    doi: 11.3969/j.issn.1674-8115.2015.01.021

    Abstract ( 563 )   PDF (674KB) ( 1022 )  

    Objective To explore the spatial distribution characteristics of birth defects in China and to provide scientific evidences for effective prevention and control of birth defects. Methods The database of incidence of birth defects from 1996 to 2012 was established. The ordinary Kriging interpolation was adopted to plot the map of incidence of birth defects in China. The spatial autocorrelation analysis, trend surface analysis, and spatial regression analysis were conducted for the incidence of birth defects. Results Global spatial autocorrelation analysis showed that the spatial aggregation of birth defects in China existed during 1996-2004 (I=0.226, P=0.000 6) and 2005-2012 (I=0.283, P=0.000 03). Local spatial autocorrelation analysis showed that Guangdong, Guangxi, and Fujian were “positive hotspot” regions and Heilongjiang, Jilin, and Liaoning were “negative hotspot” regions. Trend surface analysis showed that the incidence of birth defects increased first and then decreased from west to east and gradually increased from north to south. Spatial regression analysis showed that with the increase of latitude, the incidence of birth defects decreased. Conclusion The incidence of birth defects in China is of significantly geographical distribution. Different prevention and control measures should be established for different regions.

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    Analysis of spatial autocorrelation of HIV/AIDS in Chongqing from 2004 to 2012
    YANG Guo-jing, LI Qin, YI Juan, et al
    2015, 35 (1):  112. 
    doi: 11.3969/j.issn.1674-8115.2015.01.022

    Abstract ( 668 )   PDF (704KB) ( 837 )  

    Objective To explore whether the spatial autocorrelation exists in the incidence of HIV/AIDS in Chongqing, discuss features of the spatial distribution, and analyze the trend of variation with time for hot regions. Methods The annual incidence of HIV/AIDS of districts and counties of Chongqing from 2004 to 2012 were calculated and the global spatial autocorrelation analysis and the local spatial autocorrelation analysis were conducted. Results The annual global Moran's I indexes from 2004 to 2012 were larger than 0 (P<0.05), which indicated that positive spatial autocorrelation existed in the data. Results of the LISA analysis showed that the cluster state was high-high or low-low. Results of the G*i analysis indicated that the hot regions expanded with time. Conclusion The significant positive correlation exists in the spatial distribution of incidence of HIV/AIDS in Chongqing from 2004 to 2012. Hot regions expand with time, which indicates that the prevention and health care of HIV/AIDS in Chongqing should be reinforced.

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    Analysis of regulation of economic and public welfare objectives of public hospitals based on common agency theory
    LIU Zi-min, ZHANG Xin-zhu, MENG Tian-guang
    2015, 35 (1):  117. 
    doi: 11.3969/j.issn.1674-8115.2015.01.023

    Abstract ( 650 )   PDF (440KB) ( 883 )  

    Objective To analyze reasons of the imbalance between economic supervision and public welfare supervision of public hospitals and to propose applicable measures for solving the problem of common agency of multi-clients and balancing the economic supervision and public welfare supervision. Methods The theory of common agency of multi-clients was adopted to analyze the supervision system and objectives of supervisors. The benefit and welfare of 4 activities, i.e. supervisors act with cooperation, supervisors act simultaneously without cooperation, supervisors of economic supervision act first, and supervisors of public welfare supervision act first, were compared. Results Different action modes had different effects on achieving objectives of supervisors and total social welfare. Cooperation of both supervisors could maximize the social welfare. Simultaneous actions of supervisors without cooperation caused agents obtaining maximum benefits by taking advantage of the in-cooperation of supervisors. Sequential actions of supervisors without cooperation caused supervisors who acted first obtaining maximum benefits. Conclusion Cooperation of multi-supervisors can maximize the social welfare. The government should take measures to promote the cooperation of supervisors and improve the communication and coordination among different regulatory institutions during the course of health care reform, including self coordination, third-part coordination, and super-ministry system.

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    Effectiveness assessment of teaching method based on system dynamics for health policies
    JIANG Hao-yan, CAI Yu-yang
    2015, 35 (1):  123. 
    doi: 11.3969/j.issn.1674-8115.2015.01.024

    Abstract ( 484 )   PDF (328KB) ( 963 )  

    Objective To explore a novel teaching model of health policies in order to stimulate students' interest in learning, enhance their thinking skills, and improve the effect of classroom teaching. Methods Junior medical students of eight-year program who were enrolled in 2011 were randomly selected and divided into the experiment group (n=20) and control group (n=20). The case teaching method based on the system dynamics (SD) was adopted for the experiment group and traditional teaching method was adopted for the control group. The effect of SD-based case teaching method for health policies was analyzed and evaluated by the class observation and questionnaire survey. Results The classroom participation and results of response of the experiment group were better than those of the control group. Survey results indicated that 90% (18/20) of students of the experiment group favored the SD-based case teaching; 78% (14/18) of students believed that this teaching method would improve the learning interest; and 60% (12/20) of students thought that they obtained methods and ways of scientific decisionmaking. Conclusion SD-based case teaching method for health policies can stimulate the learning interest of students, inspire their thinking, and cultivate the awareness of scientific decision-making. It is worth being referenced and promoted for the teaching of health policies.

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    Review
    Advances of GPRC6A and its regulation function of glucose homeostasis
    DU Jing, BAO Yu-qian
    2015, 35 (1):  128. 
    doi: 11.3969/j.issn.1674-8115.2015.01.025

    Abstract ( 670 )   PDF (290KB) ( 940 )  

    The G protein coupled receptor family C, group 6, subtype A (GPRC6A) is a newly identified G protein coupled receptor, which involves in a broad range of physiological functions by combining with various ligands, such as calcium, basic amino acid, osteocalcin, and steroid, as well as the regulation of glucose homeostasis. Deletion of GPRC6A may cause disturbance of glucose homeostasis. Further study on GPRC6A and its regulation function of glucose homeostasis may identify a new therapeutic target for diabetes.

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    Application of cardiac magnetic resonance imaging in diagnosis of coronary artery disease and evaluation of prognosis
    HUANG Si-yi, PAN Jing-wei, WEI Meng
    2015, 35 (1):  132. 
    doi: 11.3969/j.issn.1674-8115.2015.01.026

    Abstract ( 596 )   PDF (325KB) ( 953 )  

    Coronary artery disease (CAD) is one of the most common diseases in China. Cardiac magnetic resonance imaging (CMR) is gradually becoming one of the effective tools for non-invasive detection of CAD because it can provide comprehensive information. This paper reviews progresses of the diagnosis of CAD and evaluation of the prognosis by CMR, including evaluation of the ventricular wall active reserve function by the large dose dobutamine stress test, evaluation of the myocardial blood flow perfusion by the myocardial first-pass perfusion imaging, and diagnosis and evaluation of myocardial infarction and chronic heart failure by the delayed enhancement scar imaging and T2-weighed CMR imaging.

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    Role of PI3K/AKT/mTOR signaling pathway in development and progress of pheochromocytoma and its combined targeted therapies
    CHENG Kang, SUN Fu-kang
    2015, 35 (1):  137. 
    doi: 11.3969/j.issn.1674-8115.2015.01.027

    Abstract ( 489 )   PDF (368KB) ( 956 )  

    PI3K/AKT/mTOR signaling pathway plays a vital role in the development and progress of pheochromocytoma. However, blocking one of the targets alone may activate the feedback loop of upstream targets and the parallel signaling pathways, which causes the resistance to PI3K/AKT/mTOR pathway inhibitors. This paper reviews the role of PI3K/AKT/mTOR signaling pathway in the development and progress of pheochromocytoma and latest research progresses of relevant targeted therapies, especially targeted therapies combined with multi-target and multi-pathway.

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    Brief original article
    Clinicopathologic analysis of peripheral primitive neuroectodermal tumors
    QI Qi, GUO Fei, WU Shi-wu
    2015, 35 (1):  142. 
    doi: 10.3969/j.issn.1674-8115.2015.01.028

    Abstract ( 794 )   PDF (488KB) ( 961 )  

    Objective To investigate the clinicopathologic and immunohistochemical characteristics of peripheral primitive neuroectodermal tumors (pPNET) and its diagnostic criteria. Methods 12 cases of pPNET were observed and analyzed by the HE staining, immmunohistochemistry, and imaging examination. Results 12 patients with pPNET presented progressively growing masses, local pain, and symptoms caused by mass compression. Imaging findings lacked specificity. Under the microscope, tumors consisted of small round cells and Homer-Wright rosettes were observed in 3 cases. Results of immunohistochemistry showed that the positive rate of CD99 was 100% (12/12). Positive rates of NSE and Syn were high, i.e. 92% (11/12) and 75%(9/12), respectively. The positive rate of Vim was 50% (6/12). Conclusion pPNET is a rare, highly malignant sarcoma. Pathological and morphological characteristics and immunohistochemistry labels are helpful for the diagnosis and differential diagnosis of pPNET.

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    Technique and method
    Development and application of metabolite batch quantitative tool
    WEI Run-min, HUANG Feng-jie, XIE Xie, et al
    2015, 35 (1):  146. 
    doi: 11.3969/j.issn.1674-8115.2015.01.029

    Abstract ( 579 )   PDF (810KB) ( 870 )  

    Objective To develop a software that can automatically draw standard curves and perform quantitative computations according to the requirements of quantitative analysis in metabolomics researches and to perform quantitative computations for clinical serum samples. Methods Quantitative computations were performed by the metabolite batch quantitative tool based on the results of metabolomics detections of clinical serum samples, including establishing standard curves, automatic and manual fitting, and quantifying metabolites in samples. Results A total of 40 standard curves of metabolites were established. Ninety clinical serum samples were quantified and their absolute concentrations were obtained. Conclusion The metabolite batch quantitative tool can perform quick quantitative computations for a large number of metabolites in batch samples during the course of metabolomics researches and can provide basic data pre-processing functions, such as data transformation and missing data filling, etc. The software is helpful for improving the automation and standardization levels of metabolite quantification, efficiently reducing human errors, and increasing the efficiency and accuracy of data analysis.

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