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    Research development |on mechanism of wound repair and correlative technique
    FANG Yong
    2009, 29 (12):  1403. 
    Abstract ( 2047 )   PDF (4040KB) ( 1766 )  

    Accompanying with the development of cell biology and molecular biology, researches on the mechanism of wound repair progress remarkably. Along with the overlap of physics, chemistry, biology and tissue engineering materials, the methods and technology derived from the theories of wound repair start to be used in clinical application. This article summarizes the research progress on the mechanism of wound healing and the technique of wound handling.

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    Photochemical tissue bonding in tissue repair
    YAO Min, WANG Ying
    2009, 29 (12):  1407. 
    Abstract ( 1831 )   PDF (2989KB) ( 1314 )  

    Suture was a traditional method for tissue repair. However, sutures used in the closure of surgical wounds can induce inflammation and lead to scarring that appears as crosshatch marks. Threads used for sutures may cause immune response and surgical infection. In this review, we introduce a novel sutureless technique for wound repair, called photochemical tissue bonding (PTB). Absorption of visible light by a photosensitizing dye initiates photochemical reactions leading to covalent protein-protein crosslinks that bridge the wound surfaces, thus producing an immediate seal between the wound surfaces. This technique has been applied in various tissues including skin, cornea, nerve, blood vessels, et al. PTB reduces inflammation resulting in less scarring in skin. PTB dramatically improves the incidences of vascular anastomotic leakage and stricture. In addition, PTB enhances the recovery of nerve function after nerve repair.

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    Original article (Basic research)
    Degradation of dicalcium phosphate dihydrate-coated Mg-Zn alloy and bone formation in vivo
    TAO Hai-rong, GU Jian-hua, HE Yao-hua, et al
    2009, 29 (12):  1410. 
    Abstract ( 1999 )   PDF (8696KB) ( 1157 )  

    Objective To investigate the degradation of dicalcium phosphate dehydrate-coated Mg-Zn alloy in vivo and bone formation. Methods Left femoral condyles were drilled in 72 New Zealand rabbits, and were randomly divided into experiment group (n=24, implanted with dicalcium phosphate dehydrate-coated Mg-Zn alloy rods), Mg-Zn alloy control group (n=24, implanted with Mg-Zn alloy rods) and poly-L-lactide acid rod group (n=24, implanted with poly-L-lactide acid rods). Serum concentrations of Mg2+ were examined 1 d pre-operation, and 1 d, 1 week, 2 weeks, 5 weeks and 10 weeks post-operation in experiment group and Mg-Zn alloy control group. Operation sites were examined by X-rays at 3, 6, 12 and 18 weeks post-operation. After X-ray examination at each time point, 6 rabbits in each group were sacrificed, and subjected to histopathological observation of live and kidney tissues by HE staining. Tissues from condyles of femur were observed by HE staining and 2, 4, 6-trinitrophenol rosein staining, and mineral apposition rate of bone was calculated. Results There was no significant difference in the concentrations of serum Mg2+ at each time point between Mg-Zn alloy control group and experiment group (P>0.05). X-ray examination revealed gas emerged near the implants 3 weeks after surgery in Mg-Zn alloy control group. However, there was no obvious histological abnormality in liver and kidney tissues. The mineral apposition rate was higher and the degradation of material was lower in experiment group than those in the other two groups. Conclusion Dicalcium phosphate dehydrate-coated Mg-Zn alloy has a favourable biocompatibility, and degrades more slowly in vivo.

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    Effects of berberine on cholesterol efflux in THP-1 macrophage derived foam cells
    LIU Xiao-yan, YAN Shi-min, GONG Hui, et al
    2009, 29 (12):  1415. 
    Abstract ( 2017 )   PDF (4521KB) ( 2692 )  

    Objective To investigate the effects of berberine on cholesterol efflux in THP-1 macrophage derived foam cells, and explore the possible mechanism. Methods HP-1 cells were induced into macrophages by phorbol myristate acetate (PMA), and were treated with acetylated low-density lipoprotein (Ac-LDL) to establish the THP-1 macrophage derived foam cell models. Foam cells were divided into blank control group and berberine (5 to 20 μmol/L) treatment groups according to the way of treatment and berberine concentrations. After treatment for 24 h, flow cytometry was employed to detect Ac-LDL aggregation, enzymic method was adopted to detect contents of cholesterol and triglyceride, scintillation counting technique was used to detect cholesterol efflux, and effects of peroxisome proliferator-activated receptor-γ (PPARγ) antagonist GW9662 pretreatment on cholesterol efflux (pioglitazone as positive control) were analysed. Besides, RT-PCR was applied to detect expression of liver X receptor α (LXRα) and ATP binding cassette transporter A1 (ABCA1) mRNA. Results Compared with blank control group, Ac-LDL aggregation and contents of cholesterol and triglyceride of foam cells in various berberine treatment groups decreased significantly (P<0.01), while cholesterol efflux increased (P<0.01) in a dose-dependent manner. After GW9662 pretreatment, there was no significant difference in cholesterol efflux between various berberine treatment groups and control group (P>0.05). Furthermore, expression of LXRα and ABCA1 mRNA of foam cells in various berberine treatment groups was higher than that in blank control group. Conclusion Berberine may increase cholesterol efflux in THP-1 macrophage derived foam cells, the mechanism of which may be associated with activation of PPARγ pathway and increase of expression of LXRα and ABCA1 mRNA.

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    Study on X-linked inhibitor of apoptosis associated factor-1 suppressing xenograft growth in nude mice with hepatocellular carcinoma
    ZHU Li-ming, TU Shui-ping, DAI Qiang, et al
    2009, 29 (12):  1419. 
    Abstract ( 2166 )   PDF (8049KB) ( 1123 )  

    Objective To investigate the inhibitory effect of X-linked inhibitor of apoptosis associated factor-1 (XAF1) on xenograft growth in nude mice with hepatocellular carcinoma. Methods The models of xenografted nude mice with human hepatocellular carcinoma cell line SMMC7721 were established. Intratumor injection was performed on three tumor sites in each group of mice (n=5) with recombinant adenovirus Ad5/F35-XAF1, control virus Ad5/F35-Null at the same infective titre or PBS of the same volume every two days for two weeks. The volumes of xenografts in all nude mice were measured every three days, and the differences between Ad5/F35-XAF1 group and the other two groups were compared. The apoptosis of tumor cells was determined by in situ end-labeling TUNEL method, the expression of XAF1 protein and microvessel density (MVD) were detected by immunohistochemistry. Results Intratumoral injection of Ad5/F35-XAF1 significantly inhibited the growth of tumor xenografts with smaller tumor size, less tumor weight and lower MVD compared with those injected with control virus Ad5/F35-Null and PBS (P<0.05 or P<0.01). However, the apoptosis index and expression of XAF1 protein in Ad5/F35-XAF1 group were significantly increased compared with the other two groups (P<0.01). Conclusion Ad5/F35-XAF1 significantly inhibits xenograft growth in nude mice with hepatocellular carcinoma, which is probably associated with the effects of XAF1 inducing hepatocellular carcinoma cell apoptosis and suppressing tumor angiogenesis.

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    Expression change of stromal cell-derived factor 1alpha in retinas after partial optic nerve injury
    PAN Dong-chao, BI Yong-yan, FENG Dong-fu, et al
    2009, 29 (12):  1424. 
    Abstract ( 1532 )   PDF (4942KB) ( 1199 )  

    Objective To investigate the changes of expression of stromal cell-derived factor 1α (SDF-1α) in retinas after partial optic nerve injury in rats. Methods Models with injury of partial optic nerve were induced in rats. Retinal tissues were collected 1,2,3,5,7,10 and 14 d after injury. Enzyme linked immunosorbent assay and Real-time quantitative PCR were employed to detect the expression of SDF-1α protein and mRNA in retinal tissues respectively in injury group (n=28), sham operated group (n=28) and normal control group (n=12). Results The expression of SDF-1α protein and mRNA in retinas was higher than that in sham operated group and normal control group at different time points after injury (P<0.01), and it reached the peak at the 5th day after injury. The expression of SDF-1α protein and mRNA maintained a high level at the 14th day after injury. Conclusion The expression of SDF-1α protein and mRNA is up-regulated after partial optic nerve injury, and may last for a long time.

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    Effect of mesenchymal stem cells on subcutaneous xenograft tumors in mice with Lewis lung cancer
    LIU Feng, JIANG Bin, ZHANG Wen-ying, et al
    2009, 29 (12):  1428. 
    Abstract ( 2055 )   PDF (3557KB) ( 1171 )  

    Objective To investigate the effect of mesenchymal stem cells (MSCs) on subcutaneous xenograft tumors in mice with Lewis lung cancer. Methods MSCs isolated from bone marrow of C57BL/6 mice were made into single cell suspension and were cultured in vitro. The cells of the 4th to 5th passage were used for the subsequent experiments. Fifty six C57BL/6 mice were inoculated subcutaneously with Lewis lung cancer cells, and  were grouped into Group D0 (MSCs were given simultaneously with inoculation)and Group D10(MSCs were given 10 d after inoculation). Group D0 included three subgroups (n=8): Group 1 with inoculation of tumor cells, Group 2 with inoculation of tumor cells and MSCs, and Group 3 with inoculation of tumor cells and tail intravenous injection of MSCs. Group D10 included four groups: Group 4 with inoculation of tumor cells and injection of MSCs in tumors, Group 5 with equivalent PBS (the control of Group 4), Group 6 with inoculation of tumor cells and tail intravenous injection of MSCs, and Group 7 with equivalent PBS (the control of Group 6). The time of tumor formation and the volume of tumor were observed and compared among the groups. Results In Group D0, earlier onset of tumor development was observed in Group 2 as compared to Group 1 and Group 3 (P<0.05), while there was no significant difference on the volume of tumor in the three groups (P>0.05). In Group D10, the volume of tumors were larger in Group 4 compared to the control (P<0.05), while there was no significant difference on the volume of tumors between Group 6 and the control (P>0.05). Conclusion Inoculating mixture of MSCs and Lewis lung cancer cells  accelerates tumor formation,and injection of MSCs in tumors stimulates the growth of tumors.

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    Relationship between extracellular matrix metalloproteinase inducer and clinical type of coronary heart disease
    ZHANG Jun-feng, WANG Chang-qian, WANG Jin, et al
    2009, 29 (12):  1431. 
    Abstract ( 1750 )   PDF (3541KB) ( 1308 )  

    Objective To investigate the relationship between extracellular matrix metalloproteinase inducer (EMMPRIN), the upstream regulatory factor of matrix metalloproteinase (MMPs), and the formation of atherosclerosis and the clinical type of coronary heart disease. Methods A total of 223 patients were classified into four groups according to results of coronary angiography (CAG) and clinical data: STEMI group (65 patients with ST-segment elevation myocardial infarction), NSTE ACS group (42 patients with non-ST-segment elevation acute coronary syndrome), SAP group (75 patients with stable angina pectoris) and normal control group (41 patients of CAG-negative). The mean fluorescence intensity (MFI) of EMMPRIN on monocytes of peripheral blood (PBMCs) were examined by flow cytometry. MMP-9 in serum was measured with ELISA; high-sensitivity C-reactive protein (hs-CRP) in serum was measured with immune velocity method. Results The EMMPRIN MFI on PBMCs in SAP group, STEMI group and NSTE ACS group was higher than that in the normal control group (P<0.05 or P<0.01). The EMMPRIN MFI in STEMI group and NSTE ACS group was higher than that in SAP group (P<0.05 or P<0.01). The expression characteristic of EMMPRIN on the PBMCs was consistent with that of hs-CRP and MMP-9 in each group. The EMMPRIN MFI of the PBMCs had positive correlation with the level of MMP-9 and hs-CRP in serum (r=0.168,P<0.05;r=0.305, P<0.01). Conclusion EMMPRIN may has promotive effect on the formation of atherosclerosis and unstablility of coronary heart disease as an upstream regulatory factor of MMPs.

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    Effects of different dialysates on apoptosis and expression of PKCdelta of U937 cell line
    ZHU Xiao-juan, GUO Li-li, PAN Yu, et al
    2009, 29 (12):  1434. 
    Abstract ( 1577 )   PDF (6597KB) ( 1261 )  

    Objective To investigate the effects of different dialysates on expression of protein kinase C-δ (PKCδ) and apoptosis of U937 cell line. Methods Different dialysates were added into culture fluid with U937 cell line at exponential phase of growth, and groups were divided: fluid A+fluid B group (dialysate A+dialysate B), fluid A+fluid B+rottlerin (PKCδ specific inhibitor)group, fluid A+powder B group (dialysate A+powder B) and fluid A+powder B + rottlerin group. Besides, blank control group and normal control group were established. Cells were harvested 24 h and 48 h after treatment, morphological changes were observed by Hoechst33258 fluorescence staining, cell apoptosis was measured by Annexin-V-FITC/PI double staining, and expression of PKCδ mRNA and protein was detected by RT-PCR and Western blotting, respectively. Results Cell apoptosis significantly increased in fluid A+powder B group, with typical morphology of apoptosis. After treatment for 24 h and 48 h, cell apoptosis rates in fluid A+powder B group were (34.99±3.54)% and (46.35±4.22)%, respectively, significantly higher than those at corresponding time points in blank control group [(7.40±1.33)% and (7.62±0.95)%], normal control group [(5.37±0.29)% and (6.46±1.02)%] and fluid A+powder B+rottlerin group [(23.58±4.12)% and (33.70±4.50)%] (P<0.05). Compared with normal control group, blank control group and fluid A+powder B+rottlerin group, the expression of PKCδ mRNA and protein of U937 cells in fluid A+powder B group were significantly increased (P<0.05). There was no significant difference in cell apoptosis rates and expression of PKCδ mRNA and protein between fluid A+fluid B group and blank control group, normal control group and fluid A+fluid B+rottlerin group (P>0.05). Conclusion Fluid A+powder B can significantly increase apoptosis of U937 cell line, the mechanism of which may be associated with the up-regulation of expression of PKCδ. Compared with fluid A+powder B, fluid A+fluid B is superior in reducing  apoptosis of peripheral blood monouclear cells.

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    Experimental study of diffusion weighted imaging in diagnosis of diffuse axonal injury
    DUAN Zhi-xin, WANG Hong-cai, WU Fang-fang, et al
    2009, 29 (12):  1439. 
    Abstract ( 1780 )   PDF (5242KB) ( 1454 )  

    Objective To observe the changes of diffusion weighted imaging (DWI) after diffuse axonal injury (DAI) in rats. Methods Models of various degrees of DAI (mild, moderate, and severe) were established in 135 SD rats by Marmarou method, and MRI examinations were performed 4, 8 and 24 h after injury. Another 8 rats were served as control group. The findings of MRI were analysed, and the apparent diffusion coefficient (ADC) values were compared among each group. Results No clear traumatic lesion was found from MRI in rats after injury. Four hours after injury, ADC values decreased in each DAI group, and there were significant differences between moderate DAI group and control group, and between severe DAI group and control group (P<0.05). Eight hours after injury, ADC values increased in each DAI group, and there was no significant difference between DAI groups and control group (P>0.05). There were significant differences in ADC values between 8 h after injury and 4 h after injury in severe DAI group (P<0.05), while there was no significant difference in moderate and mild DAI groups (P>0.05). Twenty-four hours after injury, ADC values continuously increased, especially in severe trauma group. Conclusion ADC values may reveal traumatic changes that can not be demonstrated by MRI. ADC values decrease in acute phase of DAI in rats, then increased, and the degree of variation may be related to the severity of DAI.

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    Study of diffuse axonal injury at early stage using proton magnetic resonance spectroscopy
    LI Xue-yuan, WANG Bo-cheng, FENG Dong-fu, et al
    2009, 29 (12):  1443. 
    Abstract ( 1845 )   PDF (5042KB) ( 1446 )  

    Objective To investigate the early alterations of neurometabolites in the splenium of corpus callosum by proton magnetic resonance spectroscopy (1H-MRS) in patients with diffuse axonal injury (DAI) at early stage and their prognostic value. Methods Twenty-one patients with DAI underwent 1H-MRS examination in the splenium of corpus callosum 2 to 14 d after injury, the neurometabolite alterations were evaluated and compared with those of 10 healthy subjects (normal controls). Logistic regression analysis was performed to explore the correlation among neurometabolite alterations, clinical indexes and Glasgow outcome scale (GOS) six months after injury. Results There was a significant decrease in NAA/Cr and NAA/Cho and increase in Cho/Cr in the splenium of corpus callosum in patients with DAI as compared with control group (P<0.05), and these trends tended to be more evident with the increase of injury severity. NAA/Cr and NAA/Cho in patients with poor outcomes were lower than those in patients with good outcomes (P<0.05). NAA/Cr, NAA/Cho and Cho/Cr predicted long-term outcome with 89% accuracy, and the combination with GOS provided the predictive accuracy of 94%. Conclusion 1H-MRS examination in the splenium of corpus callosum at early stage of DAI can depict brain injury severity, and is useful in predicting outcomes.

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    Correlation of tumor growth and endothelial progenitor cells entering blood induced by surgical injury
    WANG Guo-jiang, DAI Qiang, ZHANG Yan-jie, et al
    2009, 29 (12):  1447. 
    Abstract ( 1623 )   PDF (3447KB) ( 1064 )  

    Objective To explore the correlation of tumor growth and endothelial progenitor cells (EPC) entering blood induced by surgical injury in tumor bearing nude mice. Methods Forty-two tumor bearing nude mice were randomly divided into seven groups (n=6): non-surgical injury groups (1 d and 30 d), anesthetic group, surgical injury groups (24 h, 48 h, 72 h and 30 d after surgery). Blood samples and xenograft tumor tissues were taken from anesthetic group 24 h after anaesthesia and surgical injury groups 24 h, 48 h, 72 h and 30 d after surgery. EPC levels in peripheral blood were measured by flow cytometry, serum VEGF levels were determined by ELISA, microvessel density (MVD) and expression of VEGF were detected by immunohistochemistry. Results The levels of EPC in 24 h post-surgery group, 48 h post-surgery group and 72 h post-surgery group were significantly higher than that in non-surgical injury 1 d group (P<0.05). The levels of VEGF in 24 h post-surgery group, 48 h post-surgery group, 72 h post-surgery group and anesthetic group were significantly higher than that in non-surgical injury 1 d group (P<0.05). There was no significant difference in MVD among groups (P>0.05). Pearson correlation analysis revealed that serum VEGF levels were related to EPC levels in peripheral blood (r=0.695 6, P<0.01), while EPC levels in peripheral blood were not related to MVD (r=0.221 4, P>0.05), and serum VEGF levels had no correlation with MVD (r=0.224 9, P>0.05). Conclusion Surgical injury has no obvious influence on xenograft tumor growth.

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    Expression of STAT3, pSTAT3, VEGF and bFGF in tissues of nonsmall-cell lung cancer
    WANG Jiong-yi, JIANG Bin, LIU Feng, et al
    2009, 29 (12):  1450. 
    Abstract ( 2083 )   PDF (6511KB) ( 1333 )  

    Objective To investigate the expression of signal transducer and activator of transcription 3 (STAT3), phosphorylated STAT3 (pSTAT3), vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in tissues of nonsmall-cell lung cancer (NSCLC) and their relationship. Methods The expression of STAT3, pSTAT3, VEGF and bFGF in tissues of 68 cases of NSCLC and lung tissues of 27 normal cases were detected by immunohistochemistry methods, and their relationship with clinicopathological features was analyzed. Results The expression of STAT3, pSTAT3, VEGF and bFGF in tissues of NSCLC was significantly higher than that in normal lung tissues (P<0.05). There was positive correlationship among pSTAT3,VEGF and bFGF in tissues of NSCLC (P<0.05). The expression of STAT3 and pSTAT3 in NSCLC with poor differentiation was higher than that in NSCLC with high or moderate differentiation, the expression in NSCLC at TNM Ⅲ+Ⅳ stage was higher than that in NSCLC at TNMⅠ+Ⅱ stage, and the expression in NSCLC with lymph node metastasis was higher than that in NSCLC without lymph node metastasis(P<0.05). The expression of VEGF and bFGF in NSCLC at TNM Ⅲ+Ⅳ stage was higher than that in NSCLC at TNMⅠ+Ⅱ stage (P<0.05), and the expression of bFGF in NSCLC with lymph node metastasis was higher than that in NSCLC without lymph node metastasis (P<0.05). Conclusion STAT3,pSTAT3,VEGF and bFGF are highly expressed in NSCLC, and are involved in tumor metastasis and invasion.

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    Effect of Kanglaite injection on immune function of rats with Lewis lung carcinoma
    WU Yan, PAN Pei, WANG Yu-jie, et al
    2009, 29 (12):  1455. 
    Abstract ( 1780 )   PDF (4556KB) ( 1469 )  

    Objective To investigate the effect of Kanglaite injection (KLT) on immunological function of rat models with Lewis lung carcinoma. Methods Forty C57BL/6 mice were used to establish Lewis lung carcinoma models and divided randomly into the high dose(25 mL/kg), middle dose (12.5 mL/kg) and low dose (6.25 mL/kg) of KLT groups and model group(n=10). The mice in the KLT groups were sacrificed after injecting corresponding dose of KLT with intraperitoneal injection for 14 d. No treatment was performed on the rats in model group. The body weight, tumor and spleen weight was weighed, then the ratio of tumor restriction and the index of spleen was calculated. MTT colorimetric method and ELISA were used to detected activity of T cell proliferation and expression of IL-2 in spleen. The expression of NF-κB and IκBα protein was detected by Western blot. Results The ratio of tumor restriction in the high, middle, low dose of KLT groups decreased gradually. The indexes of spleen of the high and middle dose of KLT groups were higher than those in the model group (P<0.05 or P<0.01).Compared with the model group, the activity of T cell proliferation in the high, middle, low dose of KLT groups and the expression of IL-2 in the high and middle dose of KLT groups was increased significantly (P<005 or P<0.01). The expression of NF-κB protein in the nuclei of high, middle, low dose of KLT groups increased dose-dependently, and the expression of NF-κB and IκBα protein in the cytoplasm decreased dosedependently. Conclusion KLT could enhance immunological function by effecting T cell proliferation, expression of IL-2, NF-κB and IκBα, while restricting tumor growth in Lewis lung carcinoma models.

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    Effects of different doses of topiramate on expression of NCAM and GAP-43 mRNA in hippocampus of rats with epilepsy
    WU Dan-hong, ZHONG Ping, BAO Guan-shui, et al
    2009, 29 (12):  1459. 
    Abstract ( 1598 )   PDF (4307KB) ( 1151 )  

    Objective To explore the effects of different doses of topiramate (TPM) on the expression of nerve cell adhesion molecule (NCAM) and growth-associated protein 43 (GAP-43) mRNA in hippocampus of rats with epilepsy. Methods Forty-eight rats were randomly divided into normal control group, kainic acid (KA) group, 10 mg/kg TPM group, 40 mg/kg TPM group, 100 mg/kg TPM group and 400 mg/kg TPM group (n=8). The models of rats with epilepsy treated by different doses of TPM were established. The behavior of rats was observed, and the expression of NCAM and GAP-43 mRNA in hippocampus of rats was determined by Real-time PCR. Results The expression of NCAM and GAP-43 mRNA in KA group was significantly higher than that in normal control group (P<0.01), while there was no significant difference between 10 and 40 mg/kg TPM groups and KA group, that in 100 and 400 mg/kg TPM groups was significantly lower than that in KA group (P<0.01), and that in 400 mg/kg TPM group was significantly lower than that in 100 mg/kg TPM group (P<0.01). Conclusion KA can up-regulate the expression of NCAM and GAP-43 mRNA in hippocampus of rats with epilepsy. Higher dose of TPM can inhibit the expression of NCAM and GAP-43 mRNA, and the inhibitory effect is related with the dose of TPM.

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    In vitro oxalate-degrading ability of 10 strains of lactic acid bacteria for yoghourt fermentation
    ZHAO Shu-tian, ZHANG Shi-qing, GU Xin, et al
    2009, 29 (12):  1463. 
    Abstract ( 2448 )   PDF (4408KB) ( 1487 )  

    Objective To investigate the oxalate-degrading abilities of 10 strains of lactic acid bacteria for yoghourt fermentation. Methods Ten different strains of lactic acid bacteria (L.acidophilus, L.paracasei, Enterococcaceae faecium, B. lactis, B. adolescentis, B. infantis, B.longum, Lactococcus lactis subsp. Cremori, L.bulgaricus and S. thermophilus) were cultured separately in culture fluid containing 5 mmol/L oxalate. Seventy-two hours after culture, the concentrations of oxalate and lactic acid bacteria were detected. Besides, blank control cultured without lactic acid bacteria was established. Results Seventy-two hours after culture, the concentrations of all the 10 strains of lactic acid bacteria were significantly higher than those before culture (P<0.01). Compared with blank control, the concentrations of oxalate in the culture fluid with 10 strains of lactic acid bacteria decreased 72 h after culture, and were significantly different from those before culture for L.acidophilus, Lactococcus lactis subsp. Cremori, B.longum, B. adolescentis and B. lactis (P<0.05 or P<0.01). The oxalate-degrading rate of B. lactis was the highest (29.03%), and that of Enterococcaceae faecium was the lowest (0.23%). The correlation analysis revealed that there was no significant correlation between times of proliferation and oxalate-degrading rates (r=0.435 7, P=0.208 2). Conclusion All of the 10 strains of lactic acid bacteria for yoghourt fermentation have the ability of oxalate degrading, and there is no correlation between lactic acid bacteria proliferation and oxalate degradation.

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    Effects of kidney calcium oxalate calculus resistant acidophilus milk versus commercially available sacidophilus milk on urinary oxalate excretion in rats
    ZHAO Shu-tian, ZHANG Shi-qing, GU Xin, et al
    2009, 29 (12):  1467. 
    Abstract ( 1868 )   PDF (4102KB) ( 1263 )  

    Objective To compare the effects of kidney calcium oxalate calculus resistant acidophilus milk (KCOCRAM) versus commercially available acidophilus milk (CAAM) on urinary oxalate excretion in rats. Methods Fifty SD rats were randomly divided into 5 groups (n=10), and were bred with KCOCRAM (KCOCRAM group), CAAM (CAAM group), sterilized KCOCRAM (sterilized KCOCRAM group), sterilized CAAM (sterilized CAAM group) and water (blank control group), respectively for a consecutive 20 d, with 4 mL/d for each rat. The data of body weight and 24 h urinary oxalate excretion volume were obtained 1 d before breeding and every 4 d during breeding. Results During breeding, body weight and 24 h urinary oxalate excretion volume of each group increased with time. There was no significant difference in urinary oxalate excretion volume among sterilized KCOCRAM group, sterilized CAAM group and blank control group (P>0.05). The increase tendency of KCOCRAM group and CAAM group was weaker than that of the other three groups. The urinary oxalate excretion volume was significantly lower in KCOCRAM group than that in sterilized KCOCRAM group from 8 d after breeding, and that was significantly lower in CAAM group than that in sterilized CAAM group from 12 d after breeding (P<0.05). The urinary oxalate excretion volume was significantly lower in KCOCRAM group than that in CAAM group from 16 d after breeding (P<0.05). There was a positive correlationship between body weight and 24 h urinary oxalate excretion volume in each group (r=0.97-0.99, P<0.01). Conclusion Both KCOCRAM and CAAM can reduce urinary oxalate excretion in rats, and the former has a more favourable effect.

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    Effects of external substance P on scalding wound healing and neovascularization in diabetic rats
    NI Tao, FANG Yong, XU Peng, et al
    2009, 29 (12):  1471. 
    Abstract ( 1736 )   PDF (4409KB) ( 1346 )  

    Objective To evaluate the effects of external substance P (SP) on scalding wound healing and neovascularization. Methods Eighty-four Wistar rats were induced into diabetic models by intraperitoneal injection of streptozotocin (STZ), and deep partial thickness scalding wound on the back with diameter of 2 cm was prepared. Rats were randomly divided into experiment group (n=42, local injection of SP) and control group (n=42, local injection of PBS). The process of wound healing was observed, and the percentages of wound closure were calculated on D0, D1, D3, D7, D10, D14, D21 post scald. The expression of CD31, surface marker of neovascular endothelial cells, was detected within the wound sites by immunohistochemical staining, and the microvessel density was calculated. Results The percentage of wound closure in experiment group was significantly higher than that in control group on D7 post scald [(42.69±3.26) % vs (30.24±1.17)%, P<0.01]. Immunohistochemical detection revealed that the expression of CD31 and the microvessel density in experiment group were significantly higher than those in control group from D7 post scald (P<0.01). Conclusion External SP may promote scalding wound healing in diabetic rats, the mechanism of which may be associated with upregulation of expression of CD31 and acceleration of neovascularization.

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    Preventive effects of calcium antagonist on injury of renal tubular epithelial cells induced by hyperoxaluria
    LI Wen-feng, ZHANG Shi-qing
    2009, 29 (12):  1474. 
    Abstract ( 1913 )   PDF (3429KB) ( 1324 )  

    Objective To investigate the effects of hyperoxaluria on rat renal tubular epithelium intracellular calcium concentration ([Ca2+]i) and cell apoptosis, and explore the protective effects of calcium antagonist-nifedipine. Methods Sixty male SD rats were randomly divided into six groups (n=10). Rats in water-drinking group were treated with deionized water, nifedipine group with deionized water and nifedipine 10 mg·kg-1·d-1, calculiinduced group with deionized water containing 1% ethylene glycol, and three calculi-induced+nifedipineintervening groups with deionized water containing 1% ethylene glycol plus nifedipine 3, 6 and 10 mg·kg-1·d-1, respectively. Four weeks later, the 24 h oxaluria concentration was measured, the apoptosis index of renal tubular epithelial cells was detected with TUNEL method, and Ca2+ fluorescence intensity of cells of renal proximal tubules was determined by flow cytometry using Fluo-3/AM staining. Results The 24 h oxaluria concentrations in calculi-induced group and calculi-induced+nifedipine-intervening groups were higher than those in water-drinking group and nifedipine group (P<0.01). The apoptosis index and Ca2+ fluorescence intensity were significantly higher in calculi-induced group than those in water-drinking group (P<0.01). The Ca2+ fluorescence intensity in calculi-induced+nifedipine (3, 6 and 10 mg·kg-1·d-1)-intervening groups was 76.7%, 62.7% and 56.4% of calculi-induced group, respectively, with a significant dose-effect relationship (r=0.839, P<0.01). The apoptosis index of renal tubular epithelial cells was significantly correlated with Ca2+ fluorescence intensity (r=0.826, P<0.01). Conclusion Hyperoxaluria can increase apoptosis and \[Ca2+\]i concentration of renal tubular epithelial cells in rats, and nifedipine can effectively protect renal tubular epithelial cells to resist hyperoxaluria.

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    Effects and mechanism of Fructus lycii on renal calcium oxalate stone formation in rats
    ZHANG Shi-qing, LI Jian-tao, GU Xin, et al
    2009, 29 (12):  1477. 
    Abstract ( 2051 )   PDF (5598KB) ( 1513 )  

    Objective To investigate the effects and possible mechanism of Fructus lycii on renal calcium oxalate stone formation in rats. Methods Wistar rats were divided into several groups according to different stone inducer (cigarette smoking, ethylene glycol solution drinking or combination of both), either Fructus lycii infusion interference or not and different interfering concentrations (10% and 25%). Besides, a blank control group was set. After treatment for 40 d, 24 h urine was collected, and renal tissue samples were obtained. The concentrations of calcium, oxalate and citric acid in urine were measured. The deposit condition of calcium oxalate crystals in nephric tubules was observed and scored. The levels of malondialdehyde (MDA) and activity of total-superoxide dismutase (T-SOD) in renal tissues were detected. Apoptosis cells in kidney were observed with TUNEL staining, and index of apoptosis was calculated. Results Compared with blank control group, the urine calcium concentration in group of combination of cigarette smoking and ethylene glycol solution drinking were significantly higher (P<0.01), the scores of calcium oxalate crystals in renal tubules, the levels of MDA in renal tissues and the index of apoptosis of renal tubule epithelial cells in groups of ethylene glycol solution drinking and combination with smoking were higher, while the concentrations of citric acid in urine and activity of T-SOD in renal tissues were lower. Ten percent and 25% Fructus lycii infusion significantly decreased the urine concentrations of calcium in group of combination of cigarette smoking and ethylene glycol solution drinking (P<0.01), decreased the scores of calcium oxalate crystals in renal tubules, the levels of MDA in renal tissues and the index of apoptosis of renal tubule epithelial cells in groups of ethylene glycol solution drinking and combination with smoking, and increased the concentrations of citric acid in urine and activity of T-SOD in renal tissues. There was no significant dose-effect relationship between two concentrations of Fructus lycii infusion. Conclusion Fructus lycii infusion can effectively inhibit the formation of renal calcium oxalate stone in rats with smoking and/or ethylene glycol drinking by reducing the free radicals and apoptosis of renal tissue, decreasing the concentration of elements for stone formation and increasing the concentration of elements for inhibition of stone formation in urine.

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    Original article (Clinical research)
    Expression of c-met, e2f-1 and Ki-67 in tissues of gastric cancer
    WU Hong-biao, ZHANG Peng, LI Xiao-qiang, et al
    2009, 29 (12):  1482. 
    Abstract ( 1864 )   PDF (5886KB) ( 1231 )  

    Objective To detect the expression of c-met (hepatocyte growth factor receptor), e2f-1 (transcription factor) and Ki-67 in tissues of gastric cancer, explore the relationship among them, and investigate their correlationship with clinicopathological characteristics and prognosis. Methods The tissue samples of gastric cancer from 86 patients with radical resection were subjected to immunohistochemical staining, and the expression of c-met, e2f-1 and Ki-67 was detected. The relationship between the clinicopathological characteristics and the expression of c-met, e2f-1 and Ki-67, and that among the expression of c-met, e2f-1 and Ki-67 were explored by univariate and multivariate data analysis. And the relationship between prognosis and expression of c-met, e2f-1 and Ki-67 was analysed by Log rank test. Results c-met, e2f-1 and Ki-67 were widely expressed in tissues of gastric cancer. The higher expression of c-met was associated with higher expression of Ki-67, shorter survival time, upper tumor location, higher lymph node metastasis rate, higher N stage and TNM stage (P<0.05), and the lower expression of e2f-1 was associated with larger tumor diameter, higher TNM stage, deeper invasion, higher lymph node metastasis rate and higher N stage (P<0.05). The multivariate analysis revealed that depth of invasion, N stage, TNM stage and expression of Ki-67 were independent factors for positive expression of c-met, and age and survival time were independent factors for positive expression of e2f-1. Log rank test demonstrated that the factors related to survival included expression of c-met, e2f-1 and Ki-67, N stage, tumor diameter, tumor location, lymph node metastasis rate, depth of invasion and TNM stage. Conclusion c-met is an indicator for malignant behavior and poorer prognosis of gastric cancer. There is a higher expression of e2f-1 in early gastric cancer, while advanced gastric cancer may be associated with a lower expression of e2f-1.

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    Correlation between Chlamydia pneumoniae and sudden sensorineural hearing loss
    AO Hua-fei, MAO Xiao-hui, GUO Zhu-ying
    2009, 29 (12):  1487. 
    Abstract ( 1414 )   PDF (4656KB) ( 1372 )  

    Objective To explore the correlation between Chlamydia pneumoniae (Cpn) infection and sudden sensorineural hearing loss (SSHL). Methods One hundred and twenty patients with SSHL were enrolled as SSHL group, and another 120 healthy subjects were served as control group. Human peripheral blood mononuclear cells (PBMCs) were isolated, and the expression of Cpn-specific antigen (Cpn-Ag) and Cpn-DNA in PBMCs was detected by direct immunofluorescence test and PCR, respectively. Specific antibodies (IgA, IgG and IgM) to Cpn were determined by microimmunofluorescence test. Results There was no significant difference in the positive findings between direct immunofluorescence test and PCR (P>0.05). There were significant differences in the positive rates of Cpn-Ag and Cpn-IgM for acute infection between these two groups (P<0.01), and there were significant differences in the positive rates of Cpn-IgA and Cpn-IgG for chronic infection between these two groups (P<0.05). Conclusion There is a significantly higher prevalence of Cpn in patients with SSHL. Cpn infection may be a possible cause for SSHL.

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    Application of MRI diffusion tensor imaging on diagnosis of traumatic brain injury
    WANG Bo-cheng, LI mei, WU Li-zhong, et al
    2009, 29 (12):  1491. 
    Abstract ( 1646 )   PDF (4588KB) ( 1439 )  

    Objective To evaluate the value of MRI diffusion tensor imaging (DTI) on diagnosis of traumatic brain injury(TBI). Methods Twenty two patients with TBI 1 to 7 days post-injury and 14 healthy controls were studied with DTI and conventional MRI. The fractional anisotropy (FA) was quantified from different regions of interest (ROI) including the genu of corpus callosum (CC), the splenium of CC, the genu of internal capsule (IC) and the posterior limb of IC bilaterally. The FA value of the same ROI was compared between TBI group and control group, and FA value of the lesion side was compared with the mirror healthy side in TBI group. Correlations between the FA and Glasgow coma scale (GCS) in TBI patients were analyzed with Pearson linear correlation. Results Compared with control group, the FA value decreased significantly in each ROI (P<0.01). Compared with the healthy side, FA value was also significantly lower in lesion side in TBI group (P<0.01). The FA value in all the sites were not correlated with GCS in TBI group (P>0.01). Conclusion DTI is sensitive for detecting the acute traumatic injury of white matter and evaluate the degree of injury. It offers the possibility to diagnose TBI earlier and accurately.

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    Outcomes of traumatic flail chest treated by operative fixation versus conservative approach
    TENG Ji-ping, CHENG Yun-ge, NI Da, et al
    2009, 29 (12):  1495. 
    Abstract ( 2152 )   PDF (4058KB) ( 1282 )  

    Objective To compare the outcomes of traumatic flail chest with multiple injuries treated by operative fixation versus conservative approach. Methods The clinical data of 60 patients with traumatic flail chest with multiple injuries were retrospectively analysed, and the outcomes between operation group (treated by operative fixation, n=32)and non-operation group (treated by conservative approach, n=28) were compared. Results The mean time of hospital stay, ICU stay and mechanical ventilation was significantly shorter, and the prevalences of chest wall deformity, pulmonary infection, pulmonary atelectasis and respiratory failure were significantly lower in operation group than those in non-operation group (P<0.05). Six months after discharge, the pulmonary function parameters such as inspiratory capacity, forced vital capacity, forced expiratory volume in one second, peak expiratory flow, total lung capacity and maximal midexpiratory flow were significantly higher in operation group than those in non-operation group (P<0.05). Conclusion Traumatic flail chest with multiple injuries treated by operative fixation may lead to less flail chest associated complications. Operative fixation has short- and long-term benefits to flail chest.

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    Effects of chain model of kindred-like nursing on self-nursing of patients with ankylosing spondylitis
    LIU Qing-fen, LIU Qiu-ming, WANG Hai-yan
    2009, 29 (12):  1499. 
    Abstract ( 1662 )   PDF (3292KB) ( 1256 )  

    Objective To evaluate the effects of chain model of kindred-like nursing on self-nursing of patients with ankylosing spondylitis (AS). Methods Sixty patients with AS were randomized into test group and control group. Based on the therapy with same drugs, test group were given contact cards and served with continuous health care, while control group were only given routine instruction on functional exercises. Both groups were measured with Bath ankylosing spondylitis disease activity index (BASDAI) and Bath ankylosing spondylitis functional index (BASFI) before and after treatment. Results After treatment for 3 months, BASDAI and BASFI of test group were significantly improved compared with control group(P<0.05). Conclusion Chain model of kindredlike nursing improves nursing ability at home, and plays an important role in disease activity and function recovery of patients with AS.

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    Clinical study of controlled hypotension with magnesium sulphate during functional endoscopic sinus surgery
    ZHOU Wei-min, CHEN Xi-ming, HOU Rui-jiao, et al
    2009, 29 (12):  1502. 
    Abstract ( 1870 )   PDF (4724KB) ( 1257 )  

    Objective To investigate the effect and safety of administering magnesium sulphate as a technique of controlled hypotension during functional endoscopic sinus surgery. Methods Seventy-five patients arranged for selective functional endoscopic sinus surgery under general anesthesia, were randomly assigned into three groups: control group, magnesium sulphate group and nitroglycerin group(n=25). The patients of magnesium sulphate group received 10% magnesium sulphate 40 mg/kg before the induction of anaesthesia, and then 15 mg·kg-1·h-1 by continuous infusion during the operation. The same volume of5% glucose was administered to the patients in control group. Constant infusion of nitroglycerin beginning with 1 μg·kg-1·min-1 was administered to the patients in nitroglycerin group. The hemodynamic indexes, amount of bleeding, fluid intake, operation time, recovery time, quality of surgery field, muscle relaxation and adverse reaction were recorded. Concentration of Mg2+ in magnesium sulphate group were compared before and after surgery. Results The patients in the magnesium sulphate group maintained balanced heart rates during the operation, while the heart rates of patients in control group and nitroglycerin group increased significantly (P<0.01). The patients of magnesium sulphate group had lower amount of bleeding and shorter operation time compared with control group (P<0.01). The quality of surgery field in magnesium sulphate group was significantly better than that in control group (P<0.01). Time for 75% recovery of myokymia and recovery index in magnesium sulphate group was higher than those in control group and nitroglycerin group(P<0.01). No adverse reaction was observed in all groups. Conclusion Magnesium sulphate can be effective and safe for controlled hypotension during functional endoscopic sinus surgery.

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    Efficacy of etanercept on ankylosing spondylitis
    ZHAO Fu-tao, ZHAO Hao, WANG Yan-ling
    2009, 29 (12):  1506. 
    Abstract ( 1641 )   PDF (3010KB) ( 1506 )  

    Objective To evaluate the efficacy and safety of etanercept in treatment of ankylosing spondylitis. Methods Eighty-six patients were randomized into experiment group and control group (n=43). Patients in experiment group were treated with non-steroidal antiinflammatory drugs (NSAIDs) and etanercept (25 mg, twice/week, subcutaneous injection in the upper arm), and those in control group were managed with NSAIDs and sulfasalazine. Reexaminations were performed after treatment for 3 months, and the prevalence of low back pain, time of morning stiffness, Bath ankylosing spondylitis disease activity index(BASDAI), Bath ankylosing spondylitis functional index(BASFI), erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) were analysed, and the prevalence of adverse effects was observed. Results Compared with those at administration, the prevalences of low back pain significantly reduced (P<0.05 or P<0.01), the time of morning stiffness significantly decreased (P<0.05), BASDAI and BASFI significantly improved, and ESR and CRP significantly decreased 3 months after treatment in both groups (P<0.05). After treatment for three months, the clinical parameters in experiment group were more favourable than those in control group (P<0.05). There was no significant difference in the prevalence of adverse effects between two groups (P>0.05). Conclusion Etanercept has favourable short-term therapeutic effects on ankylosing spondylitis with minor adverse effects.

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    Therapeutic effects of different doses of recombinant human tumor necrosis factor-receptor Ⅱ: IgG Fc fusion protein on rheumatoid arthritis
    ZHAO Hao, ZHAO Fu-tao, WANG Yan-ling
    2009, 29 (12):  1509. 
    Abstract ( 1975 )   PDF (3071KB) ( 1271 )  

    Objective To observe the therapeutic effects of different doses of recombinant human tumor necrosis factor-receptor Ⅱ:IgG Fc fusion protein (rhTNFR:Fc) on rheumatoid arthritis (RA). Methods Seventy-six patients with RA were randomly divided into rhTNFR:Fc treatment group A (n=22; rhTNFR:Fc 25 mg, twice/week and methotrexate 10 mg/week), rhTNFR:Fc treatment group B (n=24; rhTNFR:Fc 12.5mg, twice/week and methotrexate 10 mg/week) and control group (n=30; methotrexate 10 mg/week). Joint function related parameters including number of swollen joints, number of tenderness, pain degree, time of morning stiffness, Health Assessment Questionnaire (HAQ) score, erythrocyte sedimentation rate, C-reactive protein and rheumatoid factor were detected before and 2, 4, 6, 8, 12, 16 and 24 weeks after treatment in all the patients. After being treated for 3 months, the changes of joint function related paramters were evaluated and compared among groups. ACR20, ACR50 and ACR70 criteria were employed to evaluate the therapeutic effects. Results There was no significant difference in joint function related parameters among three groups before treatment (P>0.05), while those improved significantly after treatment for 3 months (P<0.05), with more favorable results in rhTNFR:Fc treatment group A and B than in control group (P<0.05). The percent of patients with ACR50 in rhTNFR:Fc treatment group A was significantly higher than that in rhTNFR:Fc treatment group B 2 and 4 weeks after treatment (P<0.05), while there was no significant difference in total effect between these two groups 8, 12, 16 and 24 weeks after treatment(P>0.05). Conclusion Small dose of rhTNFR:Fc in combination with methotrexate may yield effective and economical results for treatment of RA.

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    Application of association rules to risk prediction of sudden deafness
    CHENG Xue-feng, AO Hua-fei, GU Jian, et al
    2009, 29 (12):  1512. 
    Abstract ( 1513 )   PDF (3187KB) ( 1084 )  

    Objective To apply data mining to risk prediction of sudden deafness, and form the association rules. Methods The clinical data of 517 patients with sudden deafness was collected,  including the characteristics of 19 attributes: sex, age, season, hypertension, diabetes, heart disease, hypercholesterolemia, atherosclerosis, long-term smoking, alcoholism, mental tension, insomnia, weakness, bedridden, infection, congenital malformation, trauma, tumour and autoimmune diseases. The source database were cleaned, then mapped for mining database. Minimum support to 0.1 and minimum confidence level to 0.9 were set for analysis of association rules. Results One hundred and six strong association rules were formed, and the rules contained the relation between the incidence of sudden deafness and the characteristics of 19 attributes. Conclusion This method is conducive to make the abstract theory of mathematical statistics into useful association rules to guide the practice of disease prevention and control.

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    Original article (Public health administration)
    Preliminary study on grading of injury from adverse drug reaction
    SHENG Hong-bin, HUANG Deng-xiao, LU Yi-yan, et al
    2009, 29 (12):  1515. 
    Abstract ( 1970 )   PDF (3385KB) ( 1366 )  

    Objective To design a quantitative criterion and grading system for injury from adverse drug reaction (ADR) in order to serve the compensation system of injury from ADR. Methods Based on the other grading system of injury cases, the independent scores and serial grades were given to the ADR injury of different organs or different levels through two turns of consultation to 27 experts from Shanghai. Results Injury from ADR was graded from stage 1 to stage 10 according to the damage degree, with 1 stand for death and 10 for slightest injury. Conclusion The grading method of ADR based on the characteristics of injury accords with the condition of China, and can provide reference for making up the compensation system of ADR injury.

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    Review
    Treatment of traumatic brain injury with hypertonic saline solutions
    LUO Wei
    2009, 29 (12):  1518. 
    Abstract ( 1343 )   PDF (4106KB) ( 1134 )  

    Hypertonic saline solutions have received more attention as effective agents for the treatment of brain damage. Studies indicated the beneficial action of hypertonic saline solutions in intracranial pressure, cerebral blood flow and neurologic outcome. This article reviews the available evidence on hypertonic saline solutions as they relate to traumatic brain injury.

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    Brief original article
    Effects of intravenous injection of immunoglobulin on NK cells in peripheral blood of MRL/lpr rats
    GUO Li-ping, JIN Hui-min
    2009, 29 (12):  1522. 
    Abstract ( 1567 )   PDF (2042KB) ( 1040 )  
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    Analysis of antibacterial drugs used in outpatients
    Wang Yu-Jie, Wang Chao, He Min, et al
    2009, 29 (12):  1524. 
    Abstract ( 1514 )   PDF (3066KB) ( 1159 )  
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    Clinical analysis on asphyxia in newborn and multiple organ dysfunction syndrome
    LV Wei, DING Guo-dong, LIN Zhen, et al
    2009, 29 (12):  1527. 
    Abstract ( 1260 )   PDF (1958KB) ( 862 )  
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    Case report
    Eight cases report of epidermoid cyst diagnosed with high-frequency ultrasound
    ZHENG Li, LIU Xiao-qing
    2009, 29 (12):  1529. 
    Abstract ( 1610 )   PDF (2623KB) ( 969 )  
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    One case report of supra-oculomotor nucleus damage after traumatic brain injury
    HU Hai-lin, WANG Jun-fang, SHI Cai-hong
    2009, 29 (12):  1530. 
    Abstract ( 1218 )   PDF (2154KB) ( 968 )  
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