Objective To investigate the expression of Interleukin 6 ( IL-6 ) and interleukin 10 (IL-10) in cerebrospinal fluid(CSF) in patients with primary central nervous system lymphoma.Methods Forty cases of primary central nervous system lymphoma ( PCNSL) , 11 cases of secondary central nervous system lymphomas( SCNS) , 20 cases of brain metastase, 20 cases of central nervous system infection and 16 cases of other nervous system disease( ONSD) were collected during the period from July 2013 to December 2015 in Fudan University Hua Shan Hospital North.Enzyme linked immunosorbent assay was used to detect the levels of IL-6 and IL-10 in serum and cerebrospinal fluid.The overall level of each group was compared using Kruskal-Wallis test.Two independent samples were compared by U Mann-Whitney test.The diagnostic value of IL-6 and IL-10 levels for PCNSL was evaluated by ROC curve.Results In PCNSL group, BM group, SCNSL group, CNSI group and ONSD group, the serum levels of IL-6 and IL-10 were not statistically significant ( H values were 4.165 and 5.368, respectively, P>0.05).IL-6 levels in CSF were significantly higher in CNSI[623.73 (1018.77-184.37) pg/ml] than those in other groups ( ZPCNSL =51.36, ZSCNSL =28.18, ZBM =51.50, ZCNSI =85.45, ZONSD =42.16, P<0.05 ) .The levels of IL-10 in CSF were significantly higher in CNSI and PCNSL than those in other groups ( ZPCNSL =74.50, ZSCNSL =34.68, ZBM =35.35, ZCNSI =72.95, ZONSD =15.66, P<0.05).The levels of IL-10 in CSF in PCNSL group[64.88(20.03-206.14)pg/ml]was significantly higher than in SCNSL group[6.28(2.78-18.87)pg/ml, Z=-3.753,P<0.05], BM group[7.30(3.72 -14.49) pg/ml, Z=-5.034,P<0.05] and ONSD group [3.14(2.931-4.20)pg/ml, Z=-5.786,P<0.05].The area under the ROC curve of IL-6 and IL-10 levels in CSF was 0.461 and 0.806 respectively for the diagnosis of PCNSL.When IL-10 was 19.62 pg/ml, the sensitivity was 77.5% ( 31/40 ) and the specificity was 70.1% ( 47/67 ) .The area under the ROC curve of CSF IL-6 and IL-10 levels was 0.861 and 0.718 respectively for the diagnosis of CNSI.When IL-6 was 155.12 pg/ml, the sensitivity was 80.0% (16/20) and the specificity was 90.8% (79/87).When IL-10 was 26.76 pg/ml, the sensitivity was 80.0% ( 16/20 ) and the specificity was 66.7% ( 58/87 ) . Conclusion The expression levels of IL-6 and IL-10 in CSF can be used as an indicator for the differential diagnosis of primary central nervous system lymphoma.

Objective To explore the correlations between quantitative parameters of dynamic contrast-enhanced MRI ( DCE-MRI) and nasopharyngeal carcinoma ( NPC) clinical stage.Methods 69 patients with NPC were selected from Oct 2013 to Oct 2015 in Shaoxing People's Hospital,39 men and 30 women,aged 39-76 years old,average 50.6 ±10.5 years old.All patients underwent clinical staging,DCE-MRI and histopathological examina-tion before treatment.Clinical stages were determined based on Chinese 2008 staging system.Value of the volume transfer constant ( Ktrans) and the extravascular extracellular space volume per unit volume of tissue ( Ve) was detec-ted by bi-compartmental pharmacokinetic model.The correlations between quantitative parameters and clinical stage were analyzed by Pearson correlation analysis.Results Mean Ktrans and Ve for NPC were (0.532 ±0.156) mL/min/100cm3 and (0.982 ±0.356),respectively.Ktrans of NPC showed negative correlation with clinical stage (r=-0.506,P=0.001),T stage (r=-0.326,P=0.014),N stage (r=-0.296,P=0.017) and M stage (r=-0.312,P=0.015),while Ve showed positive correlation with clinical stage (r=0.415,P=0.002),T stage (r=0.478,P=0.001),N stage (r=0.318,P=0.015) and M stage (r=0.346,P=0.011).Conclusion DCE-MRI quantitative parameters are significantly related to clinical stage of NPC,which has important values for clinical treat-ment and prognostic assessment.

Objective To investigate the clinical value of MR diffusion tensor imaging (DTI) in evaluating diffuse axonal injury (DAI).Methods The DTI imaging data of DAI patients and healthy volunteers were analyzed retrospectively.Fractional anisotropy (FA) values were measured in corpus callosum,etc.The serum myelin basic protein (MBP) was detected.Results Compared to the control group,the FA values were lower in genu of corpus callosum,splenium of corpus callosum,anterior limb of the internal capsule,posterior limb of the internal capsule and thalamus in DAI group (P ＜ 0.01,P ＜ 0.05).The mean FA values of two groups were negatively correlated with serum MBP levels (r =-0.755,P =0.001).Conclusions DTI can noninvasively reflect the damages of white matter fibers in DAI patients,and it has importantly clinical significance in evaluating prognosis and therapeutic effect of DAI patients.

Objective To observe the phosphorylation levels of extracellular signal-regulated kinase 5 (ERK5) in acute myocardial infarction (AMI) patients and the effects of ERK5 selective inhibitor XMD8-92 on human platelet activation in vitro,and to explore its mechanism.Methods Western blot was applied to detect the phosphorylation levels of ERK5,Akt473 and Akt308 in AMI patients (n =34) and stable angina patients (n =33,control).The effects of different concentration of XMD8-92 on human platelet aggregation induced by collagen was tested by aggregometer in vitro.The release of ATP was measured simultaneously by luciferase detection.The effects of XMD8-92 on integrin aIIbβ3 were detected by platelet spreading on immobilized fibrinogen and clot retraction.The effects of XMD8-92 on phosphorylation levels of Akt473,Akt308 PTEN370 and ERK5 were detected by Western blot.Results The levels of phosphor-Akt473,Akt308 and phosphor-ERK5 were significantly higher in AMI patients than that in control group (P＜0.05).ERK5 inhibitor XMD8-92 diminished collagen-induced platelet aggregation,ATP secretion,the average area of platelet spreading on immobilized fibrinogen and the clot retraction extent.The levels of phosphor-Akt (Ser-473/Thr-308) and phosphor-PTEN (Ser370) were significantly down-regulated in the presence of XMD8-92.Conclusions ERK5 plays a role in platelet activation in AMI process.It regulates platelet activation by regulating PTEN and Akt phosphorylation.Its specific inhibitor is hoped to be new antithrombotic drug.

Objective To observe the clinical effect of modified laryngeal mask airway combining fiberoptic bronchoscope intubation in difficult airways.Methods Forty patients,21 males and 1 9 females,aged 30-55 years,ASA physical status Ⅰ or Ⅱ,scheduled for elective general anes-thesia surgery after failure to direct laryngoscope two attempts were randomly divided into two groups,20 cases in each group.Patients in group LMA-FOB underwent intubation with modified la-ryngeal mask airway combining fiberoptic bronchoscope,and patients in group FOB underwent intu-bation with fiberoptic bronchoscope.The fiberoptic bronchoscope score,the intubation time and the rate of successful intubation at first attempt were recorded.All the patients were followed up postop-eratively for adverse effects.Results The fiberoptic bronchoscope scores (Ⅰ/Ⅱ/Ⅲ/Ⅳ:1 5/4/1/0 vs. 8/4/5/3,P <0.05)and the rate of once successful intubation (90% vs.60%,P <0.05)was signifi-cantly higher,and the intubation time [(75 ± 20)s vs.(105 ± 25 )s,P < 0.05 ]was significantly shorter in group LMA-FOB than that in group FOB.In group LMA-FOB,one patient had blood stain in the LMA and one patient felt slight sore throat.There were no significant adverse effects in the two groups.Conclusion Modified Laryngeal mask airway combining fiberoptic bronchoscope intubation in difficult airways was effective to improve the grade of the view of the larynn and the success rate of intubation and shorten the intubation time.No significant adverse effect postoperatively was reported. It is relatively safe,effective and promising in patients with difficult airway.

OBJECTIVETo establish and evaluate a BA-ELISA method for the quantitative detection of cystic fibrosis transmembrane conductance regulator (CFTR) protein.

METHODSWe deliberately selected three tables of CFTR and made the synthetic peptide be expressed in E. coli, then used the antigen to immunize rabbits to obtain the anti-CFTR polyclonal serum. After that, 96 well plates were coated with the purified antibody against CFTR. The antigen CFTR which was extracted from human sperm was detected by anti-CFTR antibody labeled with biotin, horseradish peroxidase conjugated avidin, and the substrate. The concentrations of two kinds of antibodies and the experiment parameters were optimized. Thereby, the double antibody sandwich BA-ELISA method for the quantitative detection of CFTR protein was established. Furthermore, the reproducibility, specificity and so on were evaluated by clinical specimens of sperm.

RESULTSThe optimal concentration of coated anti-CFTR IgG was 4 µg/ml, while the biotin labeled anti-CFTR IgG was 10 µg/ml; the optimal blocking buffer was 1% BSA-PBST, the optimal time of the reaction between antigen and antibody was 60 min, the optimal chromogenic time was 15 min, the intra-assay and inter-assay coefficient were 2.16%-9.23% and 2.29%-11.71% respectively; The lowest detectable limit was 0.15 ng/ml; the standard curve had a good linear correlation of R2 = 0.962.

CONCLUSIONThe BA-ELISA method for the quantitative detection of CTFR protein is successfully established, and it is demonstrated that the method has strong specificity, high sensitivity and good reproducibility. It provides the basis and evidence of the further application of the method.

Animals ; Antibodies ; Cystic Fibrosis Transmembrane Conductance Regulator ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Escherichia coli ; Humans ; Peptides ; Rabbits ; Reproducibility of Results ; Sensitivity and Specificity

Objective To explore the predictive values of the levels of procalcitonin (PCT) and thyroid hormone on the prognosis in patients with systemic inflammatory response syndrome (SIRS) complicated with euthyroid sick syndrome (ESS) and their values on differential diagnosis of ESS. Methods A total of 238 patients with SIRS hospitalized in the Emergency Department, Jiangsu Provincial Hospital of Integration of Chinese and Western Medicine, Jiangsu Branch of China Academy of Chinese Medical Sciences from July 2012 to December 2014 were divided into two groups: death group (31 cases) and survival group (207 cases), 182 patients being complicated with ESS and 56 patients without ESS. The differences in the levels of PCT, free triiodothyronine (FT3) and acute physiology and chronic health evaluation (APACHE Ⅱ) score in patients with different clinical outcomes were analyzed. The effects of the inflammatory mediators including levels of PCT, interleukin-6 (IL-6), C-reactive protein (CRP), and thyroid function such as free thyroxine (FT4) and thyroid stimulating hormone (TSH) on clinical outcomes and their predictive values on death of emergency patients with SIRS were also studied. Furthermore, the correlations between APACHEⅡscore and PCT, IL-6, CRP, thyroid hormone were analyzed. Results The level of PCT and APACHE Ⅱ score in death group were significantly higher than those in survival group [PCT (ng/L): 8.38 (13.88) vs. 1.04 (3.57), APACHEⅡscore:27.42±6.88 vs. 16.35±6.72, both P<0.01], while FT3 level was obviously lower in death group (pmol/L: 2.19±0.58 vs. 3.07±0.94, P < 0.05). No significant differences in levels of IL-6, CRP, FT4 and TSH were observed between the two groups (all P>0.05). The mortality was increased markedly in patients with higher PTC level and lower FT3 level compared with normal PCT level and normal FT3 level [18.8% (30/160) vs. 1.3% (1/78), 17.1%(31/181) vs. 0 (0/57), both P<0.05]. However, the abnormalities of IL-6, CRP, FT4 and TSH levels did not contribute to patient's mortality (all P>0.05). PCT was positively correlated with APACHEⅡscore (r>0.33, P<0.001), while FT3 was negatively correlated with APACHEⅡscore (r<-0.33, P<0.001). There were no correlations between IL-6 (r = 0.319, P < 0.001), CRP (r = 0.161, P < 0.05), FT4 (r = -0.170, P < 0.01), TSH (r = -0.057, P = 0.385), and APACHEⅡscore. The levels of PCT, IL-6 and CRP and APACHEⅡscore in patients with ESS were significantly higher than those in patients without ESS [PCT (ng/L):2.54 (5.90) vs. 0.20 (0.43), IL-6 (ng/L):98.62 (351.20) vs. 16.85 (33.60), CRP (mg/L):88.00 (110.50) vs. 25.50 (48.00), APACHEⅡscore:17.62±8.17 vs. 10.98±4.97, all P<0.01]. The cut-off values for predicting patient's death of these indexes showed by receiver operating characteristic curve (ROC curve) analysis were as follows: PCT: cut-off value ≥ 1.755 ng/L, sensitivity: 87.1%, specificity: 58.0%, area under the ROC curve (AUC): 0.802; FT3: cut-off value ≤ 2.92 pmol/L, sensitivity: 93.5%, specificity: 54.1%, AUC: 0.785;APACHE Ⅱ score: cut-off value ≥ 21.5, sensitivity: 83.9%, specificity: 88.4%, AUC: 0.920. Conclusions The levels of serum PCT, FT3 and APACHEⅡscore are prognostic factors in patients with SIRS. Meanwhile, the levels of serum PCT, IL-6, CRP and APACHE Ⅱ score should be taken into consideration in differential diagnosis of ESS in patients with SIRS.

Objective To investigate the analgesic efficacy of intrathecal injection of RC-13,a competitive kinesin superfamily protein 17 antagonist,in a mouse model of bone cancer pain.Methods Forty male C3H/HeJ mice,aged 6-8 weeks,weighing 20-25 g,were randomly divided into 5 groups (n=8 each): sham operation group (group S); bone cancer pain + 5 μl dimethyl sulfoxide (DMSO) group (group R0); bone cancer pain + 2.5 μg RC-13 group (group R1); bone cancer pain + 5 μg RC-13 group (group R2) and bone cancer pain + 10 μg RC-13 group (group R3).In groups R0-3,bone cancer pain was induced by implantation of α-min-imal essence medium (α-MEM) containing osteosarcomaNCTC 2472 cells into the intramedullary space of right femur.In group S,culture medium α-MEM containing no cancer cell was injected instead.10％ DMSO 5 μl and RC-13 2.5 μg/5 μl,5μg/5μ1 and 10 μg/5 μ1 dissolved in 10％ DMSO were injected intrathecally in groups R0-3,respectively,once a day for 3 consecutive days starting from 14th day after inoculation of the tumor cells.Pain behavior was assessed by the paw withdrawal mechanical threshold (PWMT) and spontaneous lifting times (SLTs) measured at 1 day before inoculation and at 3,5,7,10,14 days after inoculation.The same tests were also performed at 1,3,5 and 7 days after administration in groups R0-3.Results Compared with group S,PWMT was significantly decreased and SLTs were increased at 7-14 days after inoculation in the other groups (P ＜ 0.05).Compared with group R0,PWMT was significantly increased and SLTs were reduced at 1 day after administration in group R1,at 1and 3 days after administration in group R2,and at 1,3 and 5 days after administration in group R3 (P ＜ 0.05).Compared with group R1,PWMT was significantly increased and SLTs were reduced at 3 days after administration in group R2,and at 1,3 and 5 days after administration in group R3 (P ＜ 0.05).Compared with group R2,PWMT was significantly increased and SLTs were reduced at 1 and 3 days after administration in group Rs (P ＜ 0.05).Conclusion Intrathecal RC-13,a competitive kinesin superfamily protein 17 antagonist,has a good analgesic efficacy in a mouse model of bone cancer pain and the efficacy is dose-dependent.

Objective To discuss the growth suppressing, apoptosing effect of new type tumor-supressor gene-p33ING1 in stomach cancer cell strain, and to explore new strategies and methods in tumor therapy. Methods The PCDNA3/p33ING1 nuclear expressing microsome was constructed, p33ING1 and wt-p53 were implanted to human stomach cancer cell both and to evaluate the effect of p33ING1 and p53 toward stomach cancer cell and synergism between them. Results The PCDNA3/p33ING1 nuclear expressing microsome was successfully constructed. The human stomach cancer cell strain SSCG-7901 under implantation of p33ING1 and wt-p53 showed a significant decrease in cell growth, the coupling time was delayed, DNA synthetic phase was shortened and G0/G1 phase prolonged. The cell collapse increased. Conclusions Despite of the tumor-inhibiting effect and biochemical activation of p33ING1, it also plays a role with p53 gene in controling growth of stomach cancer cell, inducing cell collapse and hampering cell proliferation cycle. P33ING1 and p53 are synergistic to each other.

Coprinus comatus cultivated on cotton seed hull medium decomposed lignocellulose straggly and was high of absolute biological efficiency. Lignocellulose is the main carbon source for the fruiting stage of the fungus. There existed the positive correlation between the degradation rates of the cellulose and hemicellulose in the medium and the activities of extracellular CMCase (carboxymenthelcel lulase), FPase (filter paper cellulase) and HCase (hemicellu-lase), there also existed the positive correlation between the degradation rate of the lignin in the medium and the activity of extracellular laccase, but no correlation between the degradatio rate of the lignin in the medium and the activity of peroxidase. The activity of extracellular amylase was comparatively high at mycelial growth stage, and the protease activity peek was at teh time when the fruitbody matured.