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    Original article (Basic research)
    Construction of eukaryotic expression vector of Klotho gene and its stable expression in TCMK-1 cell line
    SHEN Yue, LU Li-ming, QIAN Ying-ying, et al
    2014, 34 (5):  567. 
    doi: 10.3969/j.issn.1674-8115.2014.05.001

    Abstract ( 888 )   PDF (995KB) ( 1040 )  

    Objective To construct the eukaryotic expression vector of the mouse Klotho gene and to acquire mouse kidney epithelial cell line TCMK-1 with stable expresses Klotho. Methods The Klotho-FLAG gene was amplified by PCR and cloned to the corresponding site of eukaryotic expression vector pCD-CXIN (CMV-MCS-IRES-Neomycin). Once confirmed by enzyme digestion and sequencing, the recombinant pCD-CXIN-Klotho plasmids were transfected to TCMK-1 cells by lipidosome and a cell line that stably expressed Klotho was obtained by G418 screening. The expression of Klotho mRNA was detected by Real-Time PCR, and the expression of Klotho protein was detected by Western blotting. Results The recombinant Klotho gene expression vector with FLAG tag was constructed, and the cell line obtained by transfection and G418 screening presents higher expression of Klotho mRNA and protein compared with that of control groups. Conclusion The recombinant Klotho expression vector has been successfully constructed and the TCMK-1 cell line with stable expression of Klotho is obtained, which might be constructive for the research on the functions of Klotho protein in mouse kidney epithelial cells.

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    Protective effect of sodium valproate on rats with adriamycin-induced nephropathy
    DAI Qin, LIU Jian, HAO Xu, et al
    2014, 34 (5):  573. 
    doi: 10.3969/j.issn.1674-8115.2014.05.002

    Abstract ( 798 )   PDF (1917KB) ( 831 )  

    Objective To investigate the protective effect of sodium valproate (VPA), an inhibitor of histone deacetylases, on rats with adriamycin-induced nephropathy. Methods Twenty male SD rats were randomly divided into the normal control group (n=6) and adriamycin-induced nephropathy group (the model was established by penial vein injection of adriamycin, n=14). The Adriamycin-induced nephropathy group was further divided into the model group (n=7) and VPA treatment group (n=7). After the model was established for eight weeks, urine samples of rats were collected by metabolism cages and blood samples were extracted from the abdominal aorta. Then rats were sacrificed and renal tissues were obtained. Changes of serum creatinine (SCr), blood urea nitrogen (BUN), triacylglycerol (TG), total cholesterol (TC), glutamic-oxal acetic transaminase (GOT), glutamic-pyruvic transaminase (GPT), albumin(Alb), and urinary albumin to creatinine ratio (UACR) of each group were observed. Changes of the collagen of tubulointerstitium and glomeruli were observed by the optical microscope after being stained by picrosirius. The expression of αsmooth muscle actin (α-SMA) in renal tissues was detected by the immunohistochemistry. The expressions of collagen typeⅠ (ColⅠ), collagen type Ⅲ (ColⅢ), and α-SMA mRNA in renal tissues were determined by the Real-Time PCR. Results Compared to the normal control group, SCr, BUN, TG, TC, and UACR of adriamycin-induced nephropathy group were significantly increased (P<0.05 and P<0.01); Alb was significantly decreased (P<0.01); and the expressions of collagen and α-SMA in tubulointerstitium and glomeruli were increased. The differences of GOT and GPT among three groups were not statistically significant (P>0.05). Compared to the model group, SCr, BUN, TC, and UACR of the VPA treatment group were significantly decreased (P<0.05); the expressions of collagen and α-SMA in tubulointerstitium and glomeruli were decreased; and the expressions of ColⅠ, ColⅢ, and α-SMA mRNA were decreased. Conclusion VPA can significantly ameliorate the renal function of rats with, mitigate the fibrosis of renal tissues, and slow the development of the disease.

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    Effects of silencing CCL19 on migration, invasion, and proangiopoiesis of colorectal cancer cells
    LU Jun, ZHAO Jing-kun, LU Ai-guo, et al
    2014, 34 (5):  578. 
    doi: 10.3969/j.issn.1674-8115.2014.05.003

    Abstract ( 950 )   PDF (2860KB) ( 953 )  

    Objective To investigate the role of chemokine CCL19 in colorectal carcinoma tissues. Methods SW620 Cell line with highest expression level of CCL19 was screened for the study by the Real-Time quantitative PCR and Western blot. CCL19 of SW620 was silenced by siRNA. The changes of proliferation, migration and invasion, and proangiopoiesis abilities of SW620 were detected by the CCK-8, Transwell, and tube formation assay, respectively. Results The Western blotting and Real-Time PCR confirmed that the expression of CCL19 in SW620 was higher than that of other cell lines. The proliferation, migration, invasion, and proangiopoiesis abilities of SW620 cells within 48-120 h were significantly improved after CCL19 being silenced. Conclusion The expression level of CCL19 in SW620 is relatively high. Silencing CCL19 can significantly enhance the proliferation, migration, invasion, and proangiopoiesis abilities of SW620 cells in vitro, so it is believed that the CCL19 can inhibit the development of colorectal cancer and is potential to be applied to the research of anticancer drugs.

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    Effects of acetylation on acid tolerance response of Salmonella typhimurium
    REN Jie, SANG Yu, YAO Yu-feng, et al
    2014, 34 (5):  584. 
    doi: 10.3969/j.issn.1674-8115.2014.05.004

    Abstract ( 680 )   PDF (1577KB) ( 767 )  

    Objective To investigate the effect of acetyltransferase/deacetylase Pat/CobB on the acid tolerance response (ATR) of Salmonella typhimurium. Methods The pat gene and cobB gene of Salmonella Typhimurium, which encode acetyltransferase and deacetylase, were deleted by the λ-Red system. The growth differences were compared through the growth curves. The transcriptional levels of some genes involved in acid stress were detected by Real-Time PCR. The differences of acid tolerance response between deletion mutants and wild type strains both at the log phase and at the stationary phase were examined by the colony forming units (Cfu) counting. Results The deletion mutants of pat and cobB of Salmonella typhimurium were successfully constructed. The deletion of pat and cobB did not affect the growth of Salmonella Typhimurium and the acid tolerance response both at the log phase and at the stationary phase. Conclusion The pat/cob genes of Salmonella Typhimurium encode the acetylase/deacetylase. The mutation of pat/cob genes will not affect the growth of bacteria and will not involve in the acid tolerance response.

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    Effects of free fatty acid mixture on toll-like receptor-4 of pulmonary microvascular endothelial cells of rats and mechanism of FFA-induced inflammation response
    LI Jing, SHANG Jia-wei, LIU Xi, et al
    2014, 34 (5):  590. 
    doi: 10.3969/j.issn.1674-8115.2014.05.005

    Abstract ( 706 )   PDF (1377KB) ( 914 )  

    Objective To explore the activating effect of the free fatty acid (FFA) mixture on the toll-like receptor-4 (TLR4) of rat pulmonary microvascular endothelial cells (RRPMVECs) and the mechanism of FFA-induced inflammation response. Methods RRPMVECs cultured in vitro were prepared by FFA of different concentrations. The expression of TLR4 protein was detected by the Western blotting. RRPMVECs were then divided into the TLR4 siRNA group (transfected by TLR4 siRNA), scramble siRNA group (transfected by scramble siRNA), and negative control group. All groups were prepared by FFA of 0.1 mmol/L, LPS of 10 ng/mL, TNF-α of 5 ng/mL, and blank medium, respectively. The expressions of TLR4 mRNA and IKKβ mRNA were detected by the Real-Time PCR. The expressions of P-IκBα and NF-κB were measured by the Western blotting. The expression of inflammatory factor IL-1β in the cell culture supernatant was detected by the ELISA. Results The relative expression of TLR4 protein in RPMVECs increased significantly after being prepared by FFA of 0.1 mmol/L (P<0.05) and increased with the concentration of FFA. The relative expressions of TLR4 mRNA of the scramble siRNA group and negative control group increased significantly after being prepared by LPS and FFA (P<0.05) and did not significantly changed after being prepared by TNF-α. The expression of TLR4 mRNA of the TLR4 siRNA group did not significantly changed after being prepared by LPS, FFA, and TNF-α. The relative expressions of IKKβmRNA, p-IκBα, and IL-1β in RPMVECs of the scramble siRNA group and negative control group increased significantly after being prepared by LPS, FFA, and TNF-α (P<0.05). The interference of TLR4 siRNA could suppress the expressions of groups prepared by FFA and LPS, but did not suppress the expressions of groups prepared by TNF-α. Conclusion FFA induces the inflammation response of RPMVECs through the TLR4/IKKβ/NF-κB signaling pathway.

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    Effects of high glucose and advanced glycation end products on formation of vascular-like structure of vascular endothelial cells
    LIN Xue-song, WANG Le, QIAO Liang
    2014, 34 (5):  596. 
    doi: 10.3969/j.issn.1674-8115.2014.05.006

    Abstract ( 705 )   PDF (2255KB) ( 914 )  

    Objective To observe the effects of high concentration glucose and advanced glycation end products (AGEs) on wound closure, formation ability of vascular-like structure, and the expression of relevant regulatory factor of vascularization of vascular endothelial cells cultured in vitro. Methods Human umbilical vein endothelial cells (HUVECs) cultured in vitro were divided into the high concentration glucose group (30 mmol/L of D-glucose), AGEs group (150 mg/L of AGEs-BSA), high concentration glucose+AGEs group (30 mmol/L of D-glucose+150 mg/L of AGEs-BSA), normal group (5 mmol/L of D-glucose) according to different concentrations of glucose and AGEs-BSA added in the culture fluid, and the mannitol group (30 mmol/L of mannitol). The proliferation curve of injury model of HUVECs was detected by the electric cell-substrate impedance sensing (ECIS). The formation of vascular-like structure of HUVECs in Matrigel matrix was observed by the inverted microscope. The expressions of vascular endothelial growth factor (VEGF) and angiopoietin-2 (Ang-2) in cell culture supernatants were detected by the ELISA and the expression of Tie-2 receptor in HUVECs was observed by the fluorescence microscope. Results The results of ECIS showed that proliferation curves of HUVECs of each group were similar and differences of their slopes, which represented the healing rate, were not significant. Observations of the inverted microscope found that lengths of formed vascular-like structures of the high concentration glucose group, AGEs group, and high concentration glucose+AGEs group were significantly shorter than those of the normal group (P<0.05 or P<0.01). The results of ELISA indicated that expressions of the Ang-2 levels in cell culture supernatants of the high concentration glucose group, AGEs group, and high concentration glucose+AGEs group were significantly higher than those of the normal group and expressions of the VEGF levels were significantly lower (P<0.05). Observations of the fluorescence microscope showed that Tie-2 receptors were expressed in the cytomembrane and cytoplasm of HUVECs of the normal group and were only expressed in the nucleolus of HUVECs of the AGEs group. Conclusion High concentration glucose and/or AGEs can inhibit the formation ability of vascular-like structure of vascular endothelial cells. The mechanism may be relevant to the up-regulation of Ang-2, down-regulation of VEGF, and the differential expression of Tie-2 receptor in different locations of cells.

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    Effects of Wnt/β-catenin pathway on the mucus hypersecretion of epithelial cells of airway under the hypertonicity condition
    LI Qi, CHEN Gui-hua, ZHOU Xiang-dong
    2014, 34 (5):  603. 
    doi: 10.3969/j.issn.1674-8115.2014.05.007

    Abstract ( 767 )   PDF (1362KB) ( 814 )  

    Objective To investigate the effects of hypertonicity on the secretion of Mucin (MUC) 5AC of 16HBE cells and possible mediate effects of Wnt/β-catenin pathway. Methods The hypertonic medium was prepared by the solution of high concentration of sodium chloride and the16HBE cells were stimulated by the hypertonic medium. The 16HBE cells were divided into the negative control group cultured in normal medium and groups stimulated by the hypertonic medium for 3, 6, 9, and 12 h. The secretion volume of MUC5AC was detected by the ELISA. The expressions of Wnt family proteins of human airway, i.e. Wnt1, Wnt2, Wnt3a, Wnt4, Wnt5a, Wnt7a, Wnt10a, and Wnt10b, were detected by the Western blotting. The 16HBE cells were transfected by the specific interference RNA (siRNA) of Wnt4 and stimulated by the hypertonic medium. The secretion volumes of MUC5AC in culture supernatants were detected by the ELISA. The expressions of signaling molecules related to classic Wnt/β-catenin pathway were detected by the Western blotting. The translocation of nuclear factor (NF)-κB p65 was visualized by the cell immunofluorescence. Results The secretion volumes of MUC5AC in supernatants and cytoplasm of 16HBE cells cultured by the hypertonic medium were significantly higher than those of the control group. The secretion volumes of MUC5AC exhibited a time-dependent manner in a certain range of concentration (P<0.05). Compared to the negative control group, the expression levels of Wnt1, Wnt2, Wnt3a, Wnt5a, Wnt7a, Wnt10a, and Wnt10b of the hyperosmotic groups were not significantly different (P>0.05), while the expression levels of Wnt4 significantly increased (P<0.05). The relative expression levels of β-catenin and Cyclin D1 in cells of the hyperosmotic groups were significantly higher than those of the negative control group. The cell immunofluorescence indicated that nuclear translocation of NF-κB p65 occurred (P<0.05). The secretion volumes of MUC5AC in supernatants and cytoplasm of the hyperosmotic groups that were transfected by the Wnt siRNA were significantly lower than those of the hyperosmotic groups that were not transfected by the Wnt siRNA (P<0.05). The levels of β-catenin and CyclinD1 and the translocation of NF-κB p65 in cells were also significantly inhibited (P<0.05). Conclusion The Wnt/β-catenin signaling pathway plays an essential role in the hypersecretion of MUC5AC in 16HBE cells induced by the hypertonicity.

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    Inhibitory effect of trifluoperazine on Ishikawa cells of human endometrial carcinoma
    ZHENG Jing, SUN Xiao, WANG Jing, et al
    2014, 34 (5):  609. 
    doi: 10.3969/j.issn.1674-8115.2014.05.008

    Abstract ( 632 )   PDF (966KB) ( 906 )  

    Objective To explore the effects of trifluoperazine (TFP) on the growth and proliferation of Ishikawa cells of human endometrial carcinoma in vitro and their possible mechanisms. Methods The inhibitory effect of TFP of different concentrations on the growth of Ishikawa cells of human endometrial carcinoma was detected by the MTT assay. The half maximal inhibitory concentration (IC50) of the drug was calculated and the optimal time was selected. The effect of TFP on the cell cycle of Ishikawa cells was measured by the flow cytometer. The regulatory effect of TFP on the expressions of forkhead box transcription factor O1 (FOXO1) and Kruppellike factor 6 (KLF6) in Ishikawa cells was determined by the Real-Time PCR. Results TFP of different concentrations would inhibit the growth of Ishikawa cells. The inhibitory effect depended on the dose and time. The IC50 value of TFP was 16.56 μmol/L. The optimal time was 24 h for being treated by TFP of 20 μmol/L. After Ishikawa cells were treated by TFP of 20 μmol/L for 24 h, the proportion of S phase cells was significantly decreased (P<0.01) and the expression of KLF6 mRNA was significantly up-regulated (P<0.05). Conclusion The intervention of TFP in vitro can inhibit the growth and proliferation of Ishikawa cells of endometrial carcinoma. The mechanism may be relevant to up-regulate the expression of the tumor suppressor gene KLF6.

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    Study on human umbilical cord mesenchymal stem cells labeled by Effectene mediated gadopentetate dimeglumine and their MRI imaging in vitro
    SHUAI Han-lin, SHI Chang-zheng, SONG Hong, et al
    2014, 34 (5):  613. 
    doi: 10.3969/j.issn.1674-8115.2014.05.009

    Abstract ( 670 )   PDF (2738KB) ( 886 )  

    Objective To apply the human umbilical cord mesenchymal stem cells (hUCMSCs) labeled by Effectene mediated gadopentetate dimeglumine (Gd-DTPA) and to investigate the biologic characteristics of magnetically labeled stem cells and the rules of MRI of Gd-DTPA labeled MSCs in vitro. Methods The hUCMSCs was separated and purified by the tissue adherent method. The biological characteristics of cells in vitro were identified after being subcultured and amplified. The hUCMSCs was marked by Effectene transfected Gd-DTPA. The proliferation ability of Gd-DTPA labeled stem cells was detected by the counting method. Then stem cells were induced to differentiate to adipocytes and osteoblasts in vitro. The effects of Gd-DTPA on the biological characteristics of hUCMSCs were observed. The clinical 1.5T MRI system was used to observe changes of the signal intensity of Gd-DTPA labeled hUCMSCs with cell passages and explore the minimum cell mass of MRI. Results Primary cells were obtained by applying the tissue adherent method for two weeks. The cells were fusiform with whirlpool-like growth. After two passages, cells were morphologically homogeneous. The flow cytometry detected that the third passage cells had high expressions of CD29, CD44, CD90, and CD105 and no expression of CD31, CD40, CD45, and HLA-DR. The cells could be induced to adipocytes and osteoblasts in vitro. Effectene could successfully transfected with Gd-DTPA into stem cells. The proliferation of labeled stem cells was not affected and the cells could be induced to differentiate towards osteoblast or adipocyte in vitro. The results of MRI scanning of Gd-DTPA labeled stem cells showed high signal on T1WI and the continuous tracing lasted about 12 d in vitro. Conclusion The tissue adherent method can effectively isolate and purify hUCMSCs. It is feasible to perform MRI tracking in vitro through hUCMSCs labeled by Effectene transfected Gd-DTPA.

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    Effects of short-term exposure to PM2.5on chronic airway inflammation and oxidative stress of passive smoking rats
    HU Jian-rong, XU Hua-jun, LI Qing-yun, et al
    2014, 34 (5):  619. 
    doi: 10.3969/j.issn.1674-8115.2014.05.010

    Abstract ( 830 )   PDF (1420KB) ( 812 )  

    Objective To explore the effects of short-term exposure to PM2.5 on the chronic airway inflammation and oxidative stress of passive smoking rats and the possible mechanisms. Methods Forty eight male Wistar rats were randomly divided into the normal control group (n=12) and the passive smoking model group (n=36). Rats of the latter group were then randomly divided into the simple passive smoking group (n=12), low dose (1.25 mg/mL) PM2.5 exposure group (n=12), and high dose (5 mg/ mL) PM2.5 exposure group (n=12) after being given passive smoking for 45 d. The pulmonary function of rats was detected at the time point of 24 h after stopping exposure to PM2.5. The bronchoalveolar lavage fluid (BALF) was collected and cells were counted and classified after being smeared and stained by Giemsa. IL-6, T-AOC, MDA, GSH-Px, and CAT in BALF were measured. The tissues of left lung lobes were excised and the histological changes and intracellular ultrastructure were observed by the optical microscopy and electron microscopy. Results Total cell count of BALF of the passive smoking model group was significantly higher than that of the normal control group (P<0.05). The percentage of neutrophils of high dose PM2.5 exposure group was significantly higher than that of the low dose PM2.5 exposure group (P<0.05). The parameters of pulmonary function of rats were significantly decreased with the increase of the exposure concentration of PM2.5 (P<0.05). Compared to the normal control group, the IL-6 and MDA levels of BALF of the passive smoking model group were significantly increased and the T-AOC, GSH-Px, and CAT levels were significantly decreased. The differences were statistically significant (P<0.05). Compared to the simple passive smoking group, the IL-6 and MDA levels of the high dose PM2.5 exposure group were significantly increased and the T-AOC, GSH-Px, and CAT levels were significantly decreased. The differences were statistically significant (P<0.05). The results of optical microscopy showed that neutrophils, lymphocytes, macrophages, and acidophilic granulocytes in bronchial walls, vessel lumens, and small airways of rats of the passive smoking model group were infiltrated and structural damages were appeared, such as significantly thinned and fused interalveolar septum, effusion and congestion in alveolar spaces, and dilated alveolar ducts. Above inflammatory process of the PM2.5 exposure group was further exacerbated. The electron microscopy showed that the ultrastructure of type Ⅱ had obvious changes. Conclusion Short-term exposure to PM2.5 can aggravate the chronic airway inflammation and oxidative stress of passive smoking rats.

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    Effects of ulinastatin on mild and severe intestinal ischemia-reperfusion injuries of mice
    LU Hua-xiang, JIA Yi-tao, YU Wei-rong, et al
    2014, 34 (5):  626. 
    doi: 10.3969/j.issn.1674-8115.2014.05.011

    Abstract ( 676 )   PDF (1888KB) ( 730 )  

    Objective To evaluate the effects of ulinastatin on mild and severe intestinal ischemia/reperfusion(I/R) injuries of mice. Methods Intestinal I/R injury models for ischemic time of 90 min (model 1) and 180 min (model 2) were established. Adult male C57BL/6 mice were randomly divided into 6 groups (n=10 each), i.e. the normal control group, sham operation group, model 1 control group, model 1 treated group, model 2 control group, and model 2 treated group. Mice of the model 1 treated group and model 2 treated group were injected i.v. with ulinastatin (16 IU/g) after the ischemia and before the reperfusion. Mice of the model 1 control group and model 2 control group were injected with the same volume of normal saline at the same period of time. The blood of inferior vena cava was collected after two hours of reperfusion. Injuries of small intestinal mucosa were observed and the degrees of pathological injury of small intestine were scored by the Chiu scoring. The activity of myeloperoxidase (MPO) was detected and serum TNF-α and IL-6 levels were determined by the ELISA. Results The structure of small intestinal mucosa of the normal control group and sham operation group was normal. The small intestinal mucosa of other groups was significantly injured and the activity of MPO and levels of serum TNF-α and IL-6 were increased. Compared to the model 1 control group, injuries of small intestinal mucosa of the model 1 treated group were significantly severer and Chiu scores were significantly higher (P<0.01). While compared to the model 2 control group, injuries of small intestinal mucosa of the model 2 treated group were significantly milder and Chiu scores were significantly lower (P<0.01). The activity of MPO and levels of serum TNF-α and IL-6 of the model 1 treated group and model 2 treated group were significantly lower than those of relevant control groups. The differences were statistically significant (P<0.01). Conclusion Ulinastatin can relieve severe I/R injuries while aggravate mild I/R injuries. This may be relevant to its anti-inflammation effect.

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    Effects of N-stearoyltyrosine on H2O2-induced oxidative-stress damage of PC12 cells
    YAO Li-yun, LIN Qi, LU Yang
    2014, 34 (5):  631. 
    doi: 10.3969/j.issn.1674-8115.2014.05.012

    Abstract ( 647 )   PDF (983KB) ( 766 )  

    Objective To investigate the effects of N-stearoyltyrosine (NsTyr) pretreatment on oxidative stress damage of PC12 cells induced by H2O2. Methods PC12 cells cultured in vitro were divided into the blank control group, H2O2 damage group (induced by H2O2 of different concentrations for 16 h), and NsTyr pretreatment group (induced by H2O2 of 100 μmol/L for 16 h after being pretreated by NsTy of different concentrations for 30 min). The apoptosis rate and ROS level were detected by the flow cytometer. The changes of expressions of Bax and Bcl-2 were determinated by the Western blotting. Results Compared to the H2O2 (100 μmol/L) damage group, the survival rates of PC12 cells of the NsTyr (5 and 10 μmol/L) pretreatment groups were significantly increased (P<0.01); the apoptotic rates of PC12 cells decreased (P<0.05); the intracellular production of ROS decreased (P<0.05, P<0.01); and the ratios of Bcl-2/Bax increased (P<0.05). Conclusion NsTyr can protect PC12 cells against H2O2-induced oxidative-stress damage; up-regulate the expression of Bcl-2; down-regulate the expression of Bax; and inhibit the apoptosis.

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    Relationship between protective effects of ginkgo biloba extract on retinal ischemia-reperfusion injury and autophagy
    YUAN Hai-hong, ZHOU Wei, WU Guo-zhong, et al
    2014, 34 (5):  635. 
    doi: 10.3969/j.issn.1674-8115.2014.05.013

    Abstract ( 682 )   PDF (2603KB) ( 893 )  

    Objective To explore the effects of pretreatment by using the ginkgo biloba extract (GBE) on cell autophagy after rat retina being injured by the ischemiareperfusion and the mechanism on which GBE can protect the retina from ischemia-reperfusion injury. Methods Rats were randomly divided into the following five groups: the normal control group, ischemia-reperfusion model group, and groups intervened by low concentration (1 mg/kg), medium concentration (3 mg/kg), and high concentration (10 mg/kg) of GBE. The retinal ischemia-reperfusion injury model was induced by perfusing the anterior chamber with saline to elevate a hydrostatic pressure of 110 mmHg (1 mmHg=0.133 kPa) for 50 min. The thicknesses of the inner plexiform layer (IPL) and inner nuclear layer (INL) of rat retinas and cell numbers of the ganglion cell layer (GCL) were measured. The apoptosis of retinal cells was detected by the terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphatebiotin nick end labeling (TUNEL). The expression variations of LC3 and Beclin1 which were related to the autophagy were detected by the immunohistochemical staining and Western blotting. Results Compared to the model group, damages of IPL, INL, GCL, and retina of intervention groups (pretreated by GBE of 3 mg/kg and 10 mg/kg) caused by the ischemia-reperfusion injury were significantly decreased (P<0.05); the apoptosis of retinal cells was decreased (P<0.01); the numbers of positive cells of LC3 and Beclin1 were increased (P<0.01); and the expressions of LC3 and Beclin1 were upregulated. Conclusion The mechanism of the protective effects of GBE on the retinal ischemia-reperfusion injury of rats may be related to the upregulation of expressions of Beclin1 and MAP1-LC3 which are related to the autophagy.

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    Effect of extremely low frequency electromagnetic fields on expression of ROCK1 and COL1A1 in human fetal sclera fibroblasts
    NIU Zong-zhen, ZHU Huang, CAI Xiao-jing, et al
    2014, 34 (5):  640. 
    doi: 10.3969/j.issn.1674-8115.2014.05.014

    Abstract ( 733 )   PDF (937KB) ( 950 )  

    Objective To investigate the effects of extremely low frequency electromagnetic fields (ELF-EMFs) on the expression of Rho-associated coiled-coil protein kinase 1 (ROCK1) and COL1A1 in human fetal sclera fibroblasts. Methods HFSFs were cultured and passaged in vitro and divided into the exposure group which was exposed to the electromagnetic field of 0.2 mT and 50 Hz for 24 h, and the control group which was not exposed to the electromagnetic field. The expressions of ROCK1 and COL1A1 mRNA were detected by the Real-Time quantitative polymerase chain reaction (Real-Time PCR) and the expression levels of ROCK1 and COL1A1 proteins were detected by the Western blotting. Results The results of the Real-Time PCR showed that compared to the control group, the expression of ROCK1 mRNA in HFSFs of the exposure group was significantly up-regulated (t=3.006, P=0.0397), while the expression of COL1A1 mRNA was significantly downregulated (t=4.225, P=0.0134). The results of the Western blotting indicated that the expression of ROCK1 protein in HFSFs of the exposure group was increased and the expression of COL1A1 protein was decreased. Conclusion ELF-EMFs can regulate the expression of ROCK1 and COL1A1 in HFSFs and thus remodel the sclera and cause the occurrence and development of myopia.

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    Inhibitory effect of 5-fluorouracil combined with Rg3 on proliferation of gastric cancer cells
    NIE Na, WU Xiao-ling, LUO Hong-chun, et al
    2014, 34 (5):  645. 
    doi: 10.3969/j.issn.1674-8115.2014.05.015

    Abstract ( 600 )   PDF (953KB) ( 968 )  

    Objective To investigate the inhibitory effect of 5-fluorouracil (5-FU) combined with Rg3 on the proliferation of gastric cancer cells. Methods The inhibitory effect of 5-FU combined with Rg3 on the proliferation of human gastric cancer cells MGC-803 and MKN-28 was investigated by the MTT assay. The in vitro tumor spheroid model was built for studying the inhibitory effect of 5-FU combined with Rg3 on the growth of tumor spheroid. The gastric xenograft tumor model of nude mice was built and mice were randomly divided into the saline group (negative control group), 5-FU group, Rg3 group, and 5-FU+Rg3 group (combined administration group). The inhibitory effect of 5-FU combined with Rg3 on the growth of xenograft tumor of nude mice was investigated. Tumor sizes of each group were measured and growth curves of tumors were plotted. Results The viability of MGC-803 cells and MKN-28 cells of the combined administration group was significantly lower than that of the other groups after being administrated by 5-FU and Rg3 for 48 h. The difference was statistically significant (P<0.01). The sizes of tumor spheroid of MGC-803 cells and MKN-28 cells of the combined administration group were significantly smaller than those of the other groups after being administrated by 5-FU and Rg3 for 7 d. The difference was statistically significant (P<0.01). The results of apoptosis tests showed that the ability of inducing apoptosis of the combined administration group was significantly stronger than that of the other groups. The difference was statistically significant (P<0.01). The results of tumor bearing nude mice tests showed that each administration group could inhibit the growth of tumors. Compared to the negative control group, the differences were statistically significant (P<0.01). The combination of 5-FU and Rg3 could prolong the median survival time of tumor bearing nude mice. The difference was statistically significant (P<0.01). Conclusion Both 5-FU and Rg3 can inhibit the growth of gastric cancer cells and the combination of 5-FU and Rg3 may be a potentially effective treatment method for gastric cancer.

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    Original article (Clinical research)
    Relationship between vascular endothelial function and levels of liver enzymes
    ZHANG Lu, LI Fa-hong, WEI Fang-fei, et al
    2014, 34 (5):  650. 
    doi: 10.3969/j.issn.1674-8115.2014.05.016

    Abstract ( 758 )   PDF (898KB) ( 945 )  

    Objective To investigate the relationship between the vascular endothelial function and the levels of liver enzymes of untreated outpatients with hypertension. Methods Four hundred outpatients who were suspected of having hypertension but not receiving antihypertensive treatments for at least two weeks were selected. The non-invasive flow-mediated dilatation (FMD) of patients was measured by the UNEXEF38. The FMD corrected for base arterial diameter (FMDc) was also calculated. The levels of serum liver enzymes were detected by the Hitachi automated Bioanalyzer 7600-020, including γ-glutamyl transpeptidase (GGT), alkaline phosphatase (ALP), and alanine transaminase (ALT). Results The simple correlation analysis showed that levels of GGT, ALP, and ALT were all have significant negative correlation with FMD and FMDc (r=-0.17 to -0.12, P<0.05). The multiple regression analysis showed that when the level of serum GGT and ALP increased by 2.7 times, FMD would decreased by 0.50% and 1.05% (P<0.05) and FMDc would decreased by 0.53% and 1.07% (P<0.05). After confounding factors were adjusted, the level of serum ALT was not significantly correlated with the vascular endothelial function (P>0.05). Conclusion Among liver enzymes, increased serum GGT and ALP are independent risk factors for decreasing the vascular endothelial function, while the level of ALT is not correlated with the vascular endothelial function.

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    Effects of combination of modified electroconvulsive therapy on cognitive function of first-episode schizophrenia of different ages
    XU Wei-qian, QI Gang-qiao, CHEN Xing-yang, et al
    2014, 34 (5):  655. 
    doi: 10.3969/j.issn.1674-8115.2014.05.017

    Abstract ( 768 )   PDF (513KB) ( 882 )  

    Objective To investigate the effects of drug treatment alone or drug combined with modified electroconvulsive therapy (MECT) on psychotic symptoms and cognitive function of patients with schizophrenia. Methods One hundred and four inpatients with first-episode schizophrenia were divided into the low age group (age 18-26 years, n=52) and high age group (age 39-46 years, n=52) according to their ages. Each group was further divided into the drug treatment group and drug combined with MECT group based on the therapeutic method. The positive and negative syndrome scale (PANSS) score and Wisconsin card sorting test (WCST) were used to assess the psychotic symptoms and cognitive function for patients of each group before treatment and after 3, 6, and 12 weeks of treatment. The results were statistically analyzed. Results The reduction of PANSS scores of the drug combined with MECT group of low age group were significantly higher than those of the drug treatment group after three weeks of treatment. The difference was statistically significant (P<0.001). The reduction of PANSS scores of the drug combined with MECT group of high age group were significantly higher than those of the drug treatment group after three weeks and six weeks of treatment. The difference was statistically significant (P<0.001). The number of responses and correct responses and completed categories of WCST of the drug combined with MECT group of low age group were significantly higher than those of the drug treatment group after three weeks of treatment. The number of continuous errors of the drug combined with MECT group of high age group was significantly lower than those of the drug treatment group. The differences were statistically significant (P<0.05). Conclusion The MECT can rapidly improve the psychotic symptoms and increase the cognitive function of patients of different ages.

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    Meta analysis of level of peripheral triglyceride of patients with depression
    CHEN Yu, ZHOU Chan-juan, ZHOU Lin-ke, et al
    2014, 34 (5):  660. 
    doi: 10.3969/j.issn.1674-8115.2014.05.018

    Abstract ( 633 )   PDF (5179KB) ( 952 )  

    Objective To compare the level of peripheral triglyceride between patients with major depressive disorder (MDD) and healthy control (HC) subjects by the meta analysis. Methods Case-control studies or cross-sectional studies relevant to patients with depression were retrieved from the PubMed, EMBASE, China National Knowledge Internet, Wanfang Data, VIP, and SinoMED. The deadline of retrieval time was October, 2013. The analysis was conducted by the Review Manager 5.0. Results Twenty-one papers from home and abroad were selected, which included 1 767 patients with depression and 1 950 healthy control subjects. The results of meta analysis indicated that triglyceride levels of patients with depression were 15.95 (95% confidence interval: 8.87-20.03) mg/dL (1 dL=100 mL), which were significantly higher than those of healthy control subjects (P<0.000 01). The subgroup analysis showed that antidepressants, age, and body mass index had certain effects on the triglyceride level. Conclusion The level of peripheral triglyceride of patients with depression elevates significantly and may be the risk factor of metabolic syndrome.

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    Significance of echocardiographic screening of cardiac defects for fetuses with single umbilical artery
    XIN Tian-tian, XI Li-li, ZHAO Li-jiao, et al
    2014, 34 (5):  666. 
    doi: 10.3969/j.issn.1674-8115.2014.05.019

    Abstract ( 755 )   PDF (536KB) ( 938 )  

    Objective To explore the relationship between the single umbilical artery (SUA) and cardiac defects and the necessity of routinely conducting fetal echocardiography for fetuses with SUA. Methods The routine fetal echocardiography was performed for 666 singleton pregnancies with high risk of fetal congenital heart disease (CHD) from September, 2009 to September, 2012. The sequential segmental analysis method was used to scan cardiac planes and to analyze and evaluate the position, major structures, and connections of the fetal heart. All cases were divided into the isolated SUA group (group A, n=63), SUA with other high risk factors of CHD group (group B, n=15), and non-SUA with other high risk factors of CHD group (group C, n=588). The echocardiography was performed for all fetuses after they were born for one month. The cardiac defects of three groups were analyzed and compared. Results The incidences of cardiac defects of three groups were 9.5% (6/63), 40.0% (6/15), and 3.9% (23/588), respectively. The results of statistical analysis showed that the incidence of cardiac defects of group B was significantly higher than that of other two groups and the incidence of cardiac defects of group A was significantly higher than that of group C. The differences were statistically significant (P<0.05). Conclusion The incidence of CHD of fetuses with SUA is extremely high. The routine fetal echocardiography is necessary for fetuses with SUA, especially for those associated with other high risk factors of CHD.

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    Analysis of dietary nutrition status of pre-school children in suburb of Chongqing
    PENG Rong, WEI Xiao-ping, LIANG Xiao-hua, et al
    2014, 34 (5):  671. 
    doi: 10.3969/j.issn.1674-8115.2014.05.020

    Abstract ( 699 )   PDF (737KB) ( 1021 )  

    Objective To analyze the dietary nutrition status of pre-school children in suburb of Chongqing and to provide scientific basis for further nutrition intervention. Methods Three of seven kindergartens in Banan district of Chongqing were randomly selected to conduct the dietary questionnaire survey which covered general information on health, illness, and the dietary intakes of children for three consecutive days. Totally 492 valid questionnaires were returned. Results The ratios of actual intakes of staple foods, fruits and nuts, vegetables, eggs, fish and shrimp, poultry meat, beans and soy products, and milk and dairy products per child per day and low limits of those of recommended intakes were 118.1%, 135.0%, 61.7%, 77.8%, 41.7%, 281.9%, 34.7%, and 119.2%, respectively. The ratios of actual intakes of daily energy, protein, vitamin A, calcium, iron, and zinc per child per day and those of recommended intakes were 100.8%, 120.2%, 121.2%, 65.7%, 126.8%, and 75.0%, respectively. Conclusion The intakes of staple foods, fruit, and milk of children from kindergartens in suburb of Chongqing were reasonable, while the intakes of vegetables, fish and shrimp, eggs, and beans were inadequate, and the intakes of poultry meat were excessive. Intakes of nutrients were high or low except the energy and thiamin. Propaganda and education of scientific and reasonable dietary nutrition intervention for parents should be enhanced.

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    Observation on diagnosis and treatment of new-onset herpes simplex virus keratitis after cataract surgery
    GU Zhen-sheng, LU Shang-lin
    2014, 34 (5):  677. 
    doi: 10.3969/j.issn.1674-8115.2014.05.021

    Abstract ( 645 )   PDF (530KB) ( 906 )  

    Objective To investigate the clinical manifestations, diagnosis, and therapy of new-onset herpes simplex virus (HSV) keratitis after the cataract surgery and to provide references for identification and prophylaxis of this disease. Methods Seven cases with HSV keratitis after the cataract surgery from 2008 to 2012 were retrospectively analyzed. The history and process of diagnosis and treatment were traced and the possible reasons of this disease were analyzed. Results After being treated by the antivirus, low concentration steriods, artificial tears, and corneal epithelium repair agents, conditions of seven patients were much better within one week. Ulcers of four patients healed after two weeks and ulcers of other patients healed after three weeks. Only one patient's visual acuity was affected due to residual scar opacity at central region and all other patients' visual acuity reached the premorbid level. Conclusion Ocular surgical trauma followed by topically administrating corticosteriod can induce the occurrence of HSV keratitis. Since its initial clinical representations are not typical, misdiagnosis can be made.

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    Meta analysis of relationship between polymorphism of genes of vascular endothelial growth factor and advanced retinopathy of prematurity
    ZHU Tian-wen, ZHANG Yong-hong, CHEN Yan, et al
    2014, 34 (5):  681. 
    doi: 10.3969/j.issn.1674-8115.2014.05.022

    Abstract ( 791 )   PDF (1790KB) ( 754 )  

    Objective To investigate the relationship between the gene polymorphism of vascular endothelial growth factor (VEGF) and the advanced retinopathy of prematurity (ROP). Methods Retrievals of the literature in Cochrane Library, PubMed, EMBASE, CNKI, Wanfang Data, and Sino Med Database were conducted. The deadline of searches was December 30, 2013 and search words were vascular endothelial growth factor, retinopathy of prematurity, and polymorphism. References of important papers were retrospectively retrieved. Randomized controlled trials (RCTs) relevant to VEGF and advanced ROP were obtained for comparative study and the quality of papers was evaluated. The heterogeneity was estimated by the State 10.0 and the meta analysis was performed according to the results of estimation. Results A total of 9 papers (9 independent studies) with 1745 samples were selected to analyze. The funnel plot showed that there was no significant publication bias. The results of meta analysis indicated that mutations of three alleles, i.e. 405 G→C (rs2010963), -460 T→C (rs833061), and 936 C→T (rs3025039), were not relevant to patients with advanced ROP. Their OR values were 0.95 (95%CI:0.67-1.36,P=0.79), 1.20(95%CI:0.89-1.61,P=0.23), and 1.23(95%CI:0.43-3.53,P=0.7), respectively. Conclusion The polymorphism of 405 G→C (rs2010963), -460 T→C (rs833061), and 936 C→T (rs3025039) of VEGF gene are not relevant to the advanced ROP.

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    Imaging diagnosis and differential diagnosis of gastric tumors and tumor-like lesions of children
    GONG Ying, XIE Chan-lai, SUN Ying-hua, et al
    2014, 34 (5):  688. 
    doi: 10.3969/j.issn.1674-8115.2014.05.023

    Abstract ( 715 )   PDF (1526KB) ( 1036 )  

    Objective To analyze the imaging findings of the ultrasonography (US), gastrointestinal imaging (GI), computed tomography (CT), and magnetic resonance imaging (MRI) of children's primary gastric tumors and tumor-like lesions and to improve the imaging diagnosis and differential diagnosis. Methods The clinical and imaging data of seventeen children with primary gastric tumors and tumor-like lesions that were confirmed by surgeries from January, 2008 to May, 2013 were retrospectively analyzed, including seven cases of gastric duplication, two cases of gastric ectopic pancreas, two cases of inflammatory myofiroblastic tumors, two cases of immature teratoma, two cases of stromal tumors, one case of gastric diverticulum, and one case of poorly differentiated adenocarcinoma. Eleven cases were examined by the US; nine cases were examined by the GI; 15 cases were examined by the CT scan; and six cases were examined by the MRI. The imaging features of the primary gastric tumors and tumor-like lesions of children were analyzed. Results Twenty lesions occurred among seventeen cases. Among which five lesions located in the antrum; four lesions located in the fundus; nine lesions located in the gastric body (five in the greater curvature and four in the lesser curvature); and two lesions located in the gastric cardia. The average diameter of these lesions was 5.5 cm and their shapes were various. All seven cystic lesions were gastric duplications. One cystic-like lesion with air-fluid level was a gastric diverticulum. One solid-cystic mass with calcification was a teratoma. Eight solid masses were teratoma, gastric stromal tumor, inflammatory myofibroblastic tumor, adenocarcinoma, and ectopic pancreas. Conclusion The primary gastric tumors and tumor-like lesions of children are mainly cystic and solid masses. The cystic or cystic-solid masses (gastric duplication, diverticulum, and typical teratoma) have imaging characteristics while solid masses lack imaging characteristics.

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    Application value of spectral computed tomography imaging for judgment of texture of pituitary adenoma
    HOU Guo-qiang, XIE Bao-shu, JIA Feng, et al
    2014, 34 (5):  695. 
    doi: 10.3969/j.issn.1674-8115.2014.05.024

    Abstract ( 717 )   PDF (2391KB) ( 941 )  

    Objective To explore the application value of spectral computed tomography (CT) imaging for the judgment of texture of pituitary adenoma. Methods Clinical data of 36 patients treated for pituitary adenoma between December, 2011 and November, 2013 were collected. Cases were divided into the soft group, medium group, and tough group according to textures of tumors found during operations. Patients were examined by MRI and spectral CT enhanced scan before operations. Spectral attenuation curves of tumor tissues and frontal white matter and their slopes were analyzed and compared. Results Among 36 cases of pituitary adenoma, 24 cases belonged to the soft group; 8 cases belonged to the medium group; and 4 cases belonged to the tough group. Their slopes of CT spectral attenuation curves of tumor tissues were 0.669±0.066, 0.333±0.024, and 0.145±0.049, respectively. The ratios of slopes of spectral attenuation curves of tumor tissues and frontal white matter were 1.355±0.279, 0.674±0.057, and 0.318±0.005. The differences among groups were statistically significant (P<0.001). Conclusion Preoperative spectral CT imaging examination is helpful for judging the texture of pituitary adenoma. The ratio of slopes of attenuation curves of the tumor tissue and frontal white matter is relevant to the texture of pituitary adenoma.

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    Analysis of applying scoring systems of severity of illness on children with complex congenital heart diseases after operations
    ZHANG Ting-ting, ZHUANG Zhou-ying, FU Li-juan
    2014, 34 (5):  700. 
    doi: 10.3969/j.issn.1674-8115.2014.05.025

    Abstract ( 747 )   PDF (797KB) ( 869 )  

    Objective To analyze the applicability of three scoring systems of severity of illness for the evaluation of children with complex congenital heart diseases (CHD) after operations. Methods The general data of 237 patients were collected and the results of operations were scored according to the requirements of three scoring systems, i.e. the pediatric risk of mortality score (PRISM), pediatric risk of mortality score Ⅲ (PRISM Ⅲ), and pediatric clinical illness score (PCIS). The intra-class correlation coefficient (ICC) was used to measure the inter-rater reliability. The performance and applicability of the three scoring systems were evaluated by the Ttest, area under the receiver operating characteristic (ROC) curve, Hosmer-Lemeshow goodness of fit test, and missing value analysis. Results Among 237 children with complex CHD, 24 died after operations and 213 survived. ICC>0.8 meant that the collected data was reliable. Scores of PRISM and PRISM III of the death group (22.3±6.9 and 15.7±7.1) were significantly higher than those of the survival group (19.3±6.5 and 11.1±4.7) and the differences were statistically significant (P<0.05). But the difference of scores of PCIS between the death group (79.6±5.5) and survival group (81.9±7.6) was not statistically significant (P>0.05). The calibration capability of the three scores was good (P>0.05), but the discriminatory ability was insufficient. The AUCs of PRISM, PRISM III, and PCIS were 0.695, 0.724, and 0.220, respectively. Some indexes of three scoring systems were missed due to incompatibility with the work pattern of care units of our country. Conclusion The clinical applicability of PRISM, PRISM III, and PCIS for the evaluation of children with complex CHD after operations was not ideal and need to be improved. It is necessary to optimize evaluation indexes that are more sensitive to these diseases to build more specific scoring systems based on existing scoring systems.

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    ARIMA model of data of hepatitis C report of China from 2004 to 2012 and trend prediction
    WU Tian-yong, ZENG Qing, YU Meng, et al
    2014, 34 (5):  705. 
    doi: 10.3969/j.issn.1674-8115.2014.05.026

    Abstract ( 719 )   PDF (1472KB) ( 736 )  

    Objective To analyze, fit, and forecast the data of hepatitis C report of China by the autoregressive integrated moving average model (ARIMA). Methods The data of hepatitis C report of China from January, 2004 to July, 2012 was differentiated to obtain smoothness and analysis, fitting, and forecasting were conducted by the seasonal ARIMA model. Results The incidence of Hepatitis C showed an increasing trend from January, 2004 to July, 2012 and has obvious periodic changes on a basis of one year. The smoothing test, differentiation, and model identification and diagnosis were conducted based on the data of Hepatitis C report and the obtained optimal model was seasonal series of ARIMA (1,1,1)×(1,1,1)12. The residual test of this model showed a white noise sequence, fitting the data in the 95% confidence interval. Based on the cases of hepatitis C from July, 2012 to December, 2014, it could be predicted that the incidence of hepatitis C increased continuously and showed an obvious trend of periodical fluctuation. Conclusion The seasonal series ARIMA model of time series can simulate and predict the tendency of incidence of hepatitis C in China and can provide reference for the prevention and control of the epidemic.

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    Application of nuclide diuretic renography in treatment and follow-up for children with neurogenic bladder
    JIAO Xian-ting, FENG Fang, WANG Hui, et al
    2014, 34 (5):  710. 
    doi: 10.3969/j.issn.1674-8115.2014.05.027

    Abstract ( 833 )   PDF (639KB) ( 908 )  

    Objective To observe the characteristics of diuretic renography (DR) of children with neurogenic bladder (NB) and to evaluate the application value of DR in the diagnosis and follow-up of children with NB. Methods Thirteen children diagnosed as NB by clinical syndromes and urodynamic investigations underwent 15 DR examinations. Two cases were re-examined after treatment. Images and clinical data of these patients were analyzed. Images of two examinations of two followup cases were compared. Results According to the DR images, bladders of 10 cases were badly filled or even unvisualized and their ureters were wholly or partly dilatated. All patients had different degree of hydronephrosis and kidneys of 7 patients shrank (46.7%). Functions of renal cortex were severely damaged. The nephrograms indicated mechanical obstruction of upper urinary tract and some of which showed lower level. The blood perfusion rate (BPR) ratio of the atrophic kidney of patients with single atrophic kidney was significantly lower than that of the contralateral kidney (t=-11.19, P<0.05). For two follow-up cases, the functions of upper urinary tract of one case were significantly improved, while the other showed no change. Conclusion The DR imaging of children with NB showed that every part of upper urinary tract has characteristic changes, which can help the diagnosis. DR imaging can evaluate the damage level of renal functions and DR has certain application value for the evaluation of therapeutic effect and follow up.

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    Clinical manifestations and analysis of methylation of H19 imprinted genes of two cases of Silver-Russell syndrome
    Lü Yong-fen, GONG Yan, LI Pin
    2014, 34 (5):  714. 
    doi: 10.3969/j.issn.1674-8115.2014.05.028

    Abstract ( 869 )   PDF (1608KB) ( 812 )  

    Objective To investigate the genetic abnormalities of Silver-Russell syndrome, to improve the understanding of this disease, and to guide the clinical diagnosis. Methods Two patients who were suspected to have the Silver-Russell syndrome were selected and their medical history was collected in detail and summarized. The DNA of peripheral blood of patients was extracted by the genomic DNA kit and then prepared by the bisulphite conversion process. The target sequence was amplified by the PCR. The methylation status of DMR zone in upstream of H19 imprinted genes of 11P15 area was detected by the pyrophosphate sequencing. The peripheral blood of ten healthy children was collected as controls. Results The clinical characteristics and assistant examinations of two patients accorded with the clinical diagnosis of Silver-Russell syndrome. The methylation levels of six methylation sites of DMR zone in upstream of H19 imprinted genes were 16%-21% and 15%-22%. The methylation status of these areas was low. While the methylation levels of six methylation sites of controls were 48%-55%, which were significantly higher than those of patients. Conclusion The clinical manifestations of Silver-Russell syndrome are diverse. The pyrophosphate sequencing combined with the methylation analysis is easy to operate and can quantify accurately. This technology can be used for examining cases that are clinically diagnosed as SilverRussell syndrome or suspected to have the Silver-Russell syndrome.

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    Original article (Preventive medicine)
    Evaluation of intervention effect of peer-led education on sexual health of rural left-behind middle school students in Sichuan
    WANG Yang, XU Xiao-yang, CAO Hong-mei, et al
    2014, 34 (5):  719. 
    doi: 10.3969/j.issn.1674-8115.2014.05.029

    Abstract ( 683 )   PDF (850KB) ( 1044 )  

    Objective To evaluate the intervention effect of peer-led education on the sexual health of rural left-behind middle school students and explore the health education mode suitable for rural left-behind students in West China. Methods Three rural schools in Dazhou, Sichuan province were sampled by the multi-stage cluster random sampling method. Six second grade classes of each school were selected and randomly divided into the peer-led education group, traditional education group, and control group. Data of questionnaire surveys before and after the intervention were analyzed to evaluate the short term effect (knowledge). Follow-up surveys of the peer-led education group and traditional education group were conducted after being intervened for six months to evaluate the medium and longterm effect (attitude and behavior). Results The awareness rate of sexual health knowledge of the peer-led education group increased from 49.4% to 75.3% and the average sore increased from 6.77±3.14 to 8.61±2.88 after being intervened. The differences were statistically significant (P=0.001). The awareness rate of sexual health knowledge of the traditional education group increased from 51.0% to 67.5% and the average sore increased from 6.89±2.91 to 7.41±3.11 after being intervened. The differences were statistically significant (P=0.024). The difference of the average sores of the control group before and after intervention was not statistically significant (P>0.05). The average sore of the peer-led education group immediately after intervention was 8.61±2.88, which was significantly higher than 7.41±3.11 of the traditional education group (P=0.000). The difference of the average sore was 1.19 (95%CI was 0.58-1.81). Follow-up surveys after six months showed that rates of positive attitude of the peer-led education group and traditional education group increased and rates of negative behavior decreased compared to those before intervention (P<0.05). The results of the peer-led education group were better than those of the traditional education group (P<0.05). Conclusion Sexual health education for rural left-behind middle school students is effective. The peer-led education method is more suitable. Changes of health attitude and behavior still need further study.

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    Epidemiology investigation and analysis of relevant factors of adult chronic kidney disease in Yuzhong District, Chongqing
    HUANG Wen-han, TANG Lin, CAI Ying, et al
    2014, 34 (5):  725. 
    doi: 10.3969/j.issn.1674-8115.2014.05.030

    Abstract ( 640 )   PDF (685KB) ( 1099 )  

    Objective To investigate the incidence and the major risk factors of adult chronic kidney disease (CKD) in Yuzhong District, Chongqing. Methods Residents over 20 years old in Yuzhong district, Chongqing were randomly selected by the hierarchical multistage cluster sampling method and questionnaire surveys were conducted. The indicators of renal damages and the relevant risk factors were examined. The relationships between the detected results of indicators of renal damage (albuminous urine and hematuria), renal functions, and type-B ultrasonic tests and relevant risk factors (age, hypertension, diabetes mellitus, and hyperlipidemia) were analyzed by the SAS 9.2. Results A total of 3 527 residents were investigated and 3 432 of whom were examined and returned valid questionnaires. The prevalence rates of proteinuria, hematuria, and reduced renal function [eGFR<60 mL/(min·1.73 m2)] were 6.09%, 2.71%, and 3.41%, respectively. The rate of abnormal kidneys detected by the type-B ultrasonic tests was 6.59%. The detection rate of CKD of residents was 10.31% after removing duplications of positive albuminous urine, positive hematuria, reduced renal function, and abnormal kidneys detected by the type-B ultrasonic tests. The results of multiple Logistic regression analysis suggested that the age, diabetes mellitus, hypertension, and hyperlipidemia were relevant risk factors of CKD. Conclusion The prevalence of CKD of residents over 20 years old in Yuzhong District, Chongqing is similar to other large cities of China and developed countries. CKD has been a major public health problem that endangers the health of residents and attention should be paid to the early diagnose and treatment of CKD.

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    Original article (Public health administration)
    Analysis of influence factors of resilience of left-behind middle school students in Yunyang County of Three Gorges Reservoir Area by structural equation model
    GUO Xue, LIU Qin, ZHANG Fan, et al
    2014, 34 (5):  731. 
    doi: 10.3969/j.issn.1674-8115.2014.05.031

    Abstract ( 575 )   PDF (1170KB) ( 721 )  

    Objective To establish the structural equation model and explore the influence factors of resilience of left-behind middle school students in Yunyang County of the Three Gorges Reservoir Area. Methods The resilience and youth development module (RYDM) scale was used and the social demographic questionnaire was prepared to investigate 1049 left-behind middle school students in Yunyang County of the three gorges reservoir area. The statistical analysis was carried out by the structural equation model. Results The fitting index of the final structural model showed that the fitness was good and the estimation differences of all path coefficients and other parameters were statistically significant. The influence factors of resilience of left-behind middle school students were divided into external protective factors and internal protective factors. The former factors were influenced by parents who did not work at local area (β=0.024, P=0.024) and internal protective factors (β=1.136, P=0.000). The latter factors were directly influenced by the grade (β=-0.080, P=0.000), students' attitude towards parents who did not work at local area (β=-0.034, P=0.003), and whether students would worry about the safety of their parents (β=0.050, P=0.000). The external protective factors and internal protective factors of resilience were indirectly influenced by whether students were day students or not. Conclusion The resilience of left-behind middle school students in the three gorges reservoir area is influenced by many external and internal factors. Thus it is necessary to integrate individual conditions of families, schools, and students to promote the adaptation of left-behind middle school students.

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    Evaluation of implementation effect of clinical pathway for shingles
    NI Yang, WANG Su-ping, GONG Rui-jie, et al
    2014, 34 (5):  736. 
    doi: 10.3969/j.issn.1674-8115.2014.05.032

    Abstract ( 786 )   PDF (616KB) ( 867 )  

    Objective To evaluate the implementation effect of clinical pathway for shingles and to propose corresponding policies. Methods A total of 1 044 patients with shingles were selected and divided into two groups according to whether the clinical pathway was implemented, i.e. the control group (n=343) and the clinical pathway group (n=701). The differences of average hospitalization days, total hospitalization expenses per capita, and cost and ratio per unit of two groups were compared. Case interviews with relevant administrators and medical staff were also carried out. Results The major influence factors of the total hospitalization expenses were the proportion of drug expenses, nursing expenses, hospitalization days, and consultation fees (r=0.867, 0.760, 0.747, and 0.747, P=0.000). Compared to the control group, the hospitalization days of the clinical pathway group were 1.03 days fewer and the total hospitalization expense per capita was 312.68 yuan cheaper. The differences were statistically significant (P=0.003 and P=0.045). The drugs expenses, consultation fees, bed fee, and examination fees were all significantly reduced (P<0.05 or P<0.01). Administrators and medical staff generally believed that the clinical pathway benefited the doctor-patient relationship, but increased the workload. Conclusion Wider implementation and continuous improvement of clinical pathway should be fully supported and corresponding policies should be perfected. The clinical pathway forms require further elaboration and optimization and the medical resources need to be allocated reasonably. Relevant incentive measures should be made so as to improve the enthusiasm of clinical and administrative staff.

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    Study on important coefficients of posts of hospital administrative departments
    WAN Xiang-rong, XI Yi-qun, REN Yi-jiong, et al
    2014, 34 (5):  741. 
    doi: 10.3969/j.issn.1674-8115.2014.05.033

    Abstract ( 1173 )   PDF (1094KB) ( 1113 )  

    Objective To establish important coefficients for posts of hospital administrative departments and to evaluate the differences of importance of posts among departments and within each department. Methods Questionnaires were sent to 30 experts and the Delphi method was adopted to analyze the survey results. The importance of 19 posts (including the external liaison, file management, medical quality management, medical tangle management, and integrated management, etc.) of 6 departments (i.e. the Director's Office, Medical Department, Science and Education Department, Human Resource Department, Infection Control Department, and Equipment Department) was compared and evaluated. The analytical hierarchy process (AHP) method was adopted to establish the judgment matrix based on the survey results and determine the weights of indexes. Then the important coefficients of posts among administrative departments and within each department were calculated. Results Based on the quantitative analysis of the differences among evaluation indexes, the important coefficients of posts of the Director's Office, Medical Department, Science and Education Department, Human Resource Department, Infection Control Department, and Equipment Department were 1.08, 1.20, 0.96, 1.02, 0.90, and 0.84, respectively. For each department, posts with highest important coefficient were the external liaison of the Director's Office, the medical quality management of the Medical Department, the scientific research management of the Science and Education Department, the performance management of the Human Resource Department, the infected management of the Infection Control Department, and the equipment maintenance of the Equipment Department, respectively. Conclusion Important coefficients can reflect the differences of posts among administrative departments and within each department and can be used for the performance management of the administrative departments.

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    Review
    Research progress of neogenin regulation mechanism on osteogenesis
    HU Bin, CHEN Chun-yuan, WANG Yang, et al
    2014, 34 (5):  746. 
    doi: 10.3969/j.issn.1674-8115.2014.05.034

    Abstract ( 761 )   PDF (442KB) ( 845 )  

    Neogenin is a member of the DCC (deleted in colorectal cancer) family which involves in many physiological and pathological processes such as body homeostasis, tissue formation, angiogenesis, iron metabolism, immunoregulation, cell differentiation, proliferation, migration, and apoptosis through interacting with different ligands such as neuronal guidance cues (netrin) and repulsive guidance molecule(RGM). However, research on the osteogenesis modulated by neogenin is still in its infancy. This paper reviews the research progresses of neogenin, which involve its molecular characteristics, distribution, physiological functions, and roles in modulating osteogenesis through BMP signaling pathway.

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    Advances of insulin-like growth factor Ⅰ signaling pathway and therapeutic target of breast cancer
    LIANG Yue, CHEN Xiao-song, SHEN Kun-wei
    2014, 34 (5):  750. 
    doi: 10.3969/j.issn.1674-8115.2014.05.035

    Abstract ( 742 )   PDF (435KB) ( 824 )  

    The insulin-like growth factor Ⅰ (IGF-Ⅰ) signaling pathway has been found to be closely related to the therapeutic effect of breast cancer. Researches on using the IGF-Ⅰ as a new target for the target therapy of breast cancer have attracted more and more attention. Pre-clinical studies or clinical tests have been conducted for relevant targeted drugs and showed good prospect of basic researches and clinical applications. This paper reviews targeted drugs and research developments of IGF-Ⅰ.

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    Advances of relationship between E6/E7 mRNA of human papillomavirus and cervical lesions
    ZHANG Sheng-zhi, SHAO Hua-jiang, MA Jian-ting
    2014, 34 (5):  754. 
    doi: 10.3969/j.issn.1674-8115.2014.05.036

    Abstract ( 738 )   PDF (549KB) ( 863 )  

    Human papillomavirus (HPV) is closely related to the occurrence and development of cervical cancer and the expressions of carcinogenic gene E6 and E7 of the virus are the key to the carcinogenesis. When the virus gene integrates into the host, the E6 and E7 gene will be over-expressed and eventually lead to the occurrence of cervical cancer. This paper reviews the research developments of E6 / E7 mRNA, which is the transcription product of E6 and E7 gene.

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    Advances of diagnosis and catheter intervention of pulmonary atresia with intact ventricular septum
    XIE Jing, SUN Kun
    2014, 34 (5):  759. 
    doi: 10.3969/j.issn.1674-8115.2014.05.037

    Abstract ( 561 )   PDF (609KB) ( 1001 )  

    Pulmonary atresia with intact ventricular septum (PA/IVS) is a critical cyanotic congenital heart disease with high mortality in neonates and infants. Interventions must be given at early stage after birth. The survival rate and prognosis of patients with PA/IVS have been significantly improved with the development of prenatal diagnosis techniques and therapeutic techniques of medicine and surgery. This paper reviews and summarizes the progresses in diagnosis and treatment of PA/IVS to provide a reference for the clinical treatment of the disease.

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    Technique and method
    Isolation and in vitro culture of hepatic progenitor cells of normal rat liver
    ZANG Jin-feng, YUAN Yin, GAO Jun-ye, et al
    2014, 34 (5):  763. 
    doi: 10.3969/j.issn.1674-8115.2014.05.038

    Abstract ( 732 )   PDF (806KB) ( 837 )  

    Objective To explore the techniques of isolation and in vitro culture of hepatic progenitor cells of normal rat liver. Methods According to the cell surface marker CD90.1, hepatic progenitor cells of normal Wistar rats were isolated by immunomagnetic cell sorting, cultured in vitro, and induced to differentiate. Results The percentage of CD90.1 positive cells in non-parenchymal cells was (0.32±0.03) %. The flow cytometric cell sorting showed that (98.26±1.37) % of isolated cells were CD90.1 positive cells. Fresh isolated CD90.1 positive cells were strong positive for CK-19 and negative for HNF-4a. After 8 weeks of culture, these cells were negative for CK-19 and positive for HNF-4a. Conclusion Hepatic progenitor cells exist in normal rat livers and can be differentiated toward hepatocytes.

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