An online enrichment-separation system of C18 and inductively coupled plasma-mass spectrometry (ICP/MS) with flow injection （C18-FI-ICP/MS） was established for the determination of trace heavy metals in seawater. The optimized experimental parameters including pH，C18 column，concentration of methol in elute and the velocity of elute were investigated. This system was applied to the online preconcentration , separation and determination of trace heave metals in seawater samples successfully. The detection limit　(3σ) for Cd, Pb, Co, Ni and Zn was 0.03, 0.09, 0.1, 0.1, 0.3 μg/L respectively. The RSD of Cd, Pb, Co, Ni, and Zn for six measurements of seawater was 6.8%，3.4%，1.3%，2.6%，0.5%，respectively. The accuracy was verified with spiked recovery test. The recovery for five elements was 91.3%，95.1%，100.4%，100.3%，95.2%, respectively.

Objective To investigate the effects of lipoxin A4 (LXA4) on the inflammatory response to focal cerebral ischemia-reperfusion(I/R) inmy in rats.Methods Fifty-six healthy male SD rats weighing 200-250 g were randomly divided into 3 groups:group Ⅰ sham operation(group S,n=8);group Ⅱ cerebral I/R(n=24)and group Ⅲ lipoxin A4+I/R(group LXA4,n=24).Right mid-cerebral artery was occluded for 2 h by inserting cranially a nylon thread with rounded tip into internal carotid artery.LXA4 0.03 nmol/5 μl was injected into cerebral ventricle at 5 min after cerebral ischemia.Neurological deficit was scored at 24 h of reperfusion.Then four animals in each group were killed and their brains were removed for microscopic examination and expression of MPO at 24 h of reperfusion.Meantime,content of IL-1β,TNF-α,TGF-β1,and IL-10 in the brain tissue were measured at 1,6,12,24 and 48 h of reperfusion by ELISA.Glial cell activity was examined at 24 h of reperfusion by immuno-histochemistry.Results Intra-cerebroventricular administrated LXA4 0.03 nmol/5 μl provided mild neuroprotection against focal cerebral I/R injury,improved neurological deficits,and reduced morphological brain damages and PMN infiltration.LXA4 also decreased the content of TNF-α and IL-1β,and increased the content of IL-10 and TGF-β1.The numbers of activated astroglia and microglia were decreased in group LXA4 compared with group I/R.Conclusion LXA4 protects the brain against I/R injury by inhibiting inflammatory response.

Objective: To investigate the effect of survivin and bcl-2 antisense oligodeoxynucleotides ( AsODN) combined transfection on the human gallbladder carcinoma cell line GBC-SD in vitro. Methods: Survivin and Bcl-2 protein expressions were detected by immunohistochemical method; Cultured cells were divided into 4 groups: Nomal control group, survivin antisense observed group, bcl-2 antisense observed group and combined group. After transfected for 24 h, the expression of survivin mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). Cell morphological changes were observed under electron microscopy. Apoptosis index (AI) was examined by flow cytometry; Inhibitory rate (IR) was determined by the colorimetri MTT cell viability and proliferation assay. Results: Survivin and Bcl-2 protein were highly expressed in gall bladder carcinoma cells; The expression of survivin mRNA was decreased 47. 8%. Abnormal morphological changes of cells were observed in the three AsODN transfection groups; The AI in survivin antisense observed group,bcl-2 antisense observed group,and combined group was 11.38%?3.91% , 9.26%?4.15% , 28.45%?6.34% respectively and significantly higher than the nomal control group (P

Objective:To observe the expression of Th1 versus Th2 type cytokines in primary hepatic cancer(PHC)and its adjacent liver tissues.Methods:The gene expression of Th1/Th2 cytokines was detected by RT-PCR using IFN-?and IL-2 as Th1 type cytokine genes,IL-4 and IL-10 as Th2 type cytokine genes.Results:Thl type cytokines were expressed in 7 and 9 cases,while Th0 type cytokines in 4 and 2 among 11 PHC and their adjacent liver tissues,respectively.Conclusion:Th1 type cytokines are expressed predominantly in primary hepatic cancer and its adjacent liver tissue.

Objective To discuss the clinical value of D-dimer(DD) testing in diagnosis of patients with deep vein thrombosis(DVT) of the lower extremity.Methods An analysis retrospectively was made on the changes of DD testing in 106 cases of DVT at different periods after onset and different clinical stages of DVT,and DD testing in 99 patients with primary deep venous insufficiency(PDVI) as control group.(Results) DD was higher in the acute stage of DVT,and gradually decreased with time in chronic DVT,and was negative in the patients with PDVI;the positive rate was up to 85.7% in the distal DVT.Conclusions The DD testing can be used as one of the methods for diagnosis,prediction and prognosis of acute DVT,especially for diagnosis of distal DVT.

Objective To study the apoptosis of gallbladder carcinoma cell line GBC-SD induced by antisense oligodeoxynucleotide (ASODN) targeting survivin. Methods ASODN targeting survivin was transfected into GBC-SD cells mediated by lipofectin. Cultured cells were divided into 3 groups: control group,sense oligonucleotide (SODN) group and ASODN group. After transfected for 16 h, the cultured cells were harvested and the following texts were carried out. The expression of survivin mRNA was detected by RT-PCR. Flow cytometer were used to detect apoptosis. Morphological changes were observed by electron microscopy. Results The expression of survivin mRNA was decreased 47.83% in ASODN group while apoptosis was increased from (0.50?0.23)% to (26.28? 3.91)%. Abnormal morphological changes of cells were observed in ASODN group and apoptosis bodies were found in some gallbladder carcinoma cells. Conclusion The expression of survivin may be decreased in GBC-SD cells after ASODN transfection.ASODN targeting survivin could induce gallbladder carcinoma cells apoptosis effectively.

Objective To investigate the effect of lipoxin A4(LXA4) on inflammatory response in a rat model of permanent focal cerebral ischemia (PFCI). Methods Seventy-two adult male SD rats weighing 200-250 g were randomly divided into 3 groups (n = 24 each):group Ⅰ sham operation (group S); group Ⅱ PFCI and group Ⅲ LXA4. PFCI was induced by thread occlusion of right middle cerebral artery according to the method described by Longa in group Ⅱ and Ⅲ. In group Ⅲ LXA4 100 ng/5 μl was injected into right ventricle of the brain after PFCI was successfully induced, while in group Ⅰ and Ⅱ equal volume of normal saline was injected instead of LXA4. Six animals were killed at 6, 12 and 24 h of ischemia. Their brains were immediately removed for microscopic examination and determination of myeloperoxidase (MPO) activity, TNF-α, IL-10 and transforming growth factor-β1 (TGF-β1) contents in the ischemic cortex. The expression of glial fibrillary acidic protein (GFAP)was measured by immuno-histochemistry. Apoptosis in neurons was assessed using TUNEL. Results PFCI significantly increased MPO activity, TNF-α, IL-10 and TGF-β1 contents and GFAP expression in the ischemic cortex and neuronal apoptosis in group Ⅱ as compared with group S. LXA4 significanfly decreased MPO activity,TNF-α content, GFAP expression and neuronal apoptosis and increased IL-10, TGF-β1 contents at 12,24 h of ischemia. LXA4 significantly ameliorated PFCI-induced cerebral histopathologic damage. Conclusion LXA4 can protect the brain against PFCI injury by inhibiting inflammatory response.

Objective To investigate the effect of lipoxin A4 ( LXA4 ) on the permeability of blood-brain barrier (BBB) after focal cerebral ischemia-repeffnsion (I/R) in rats. Methods Fifty-four adult male SD rats weighing 200-250 g were randomly divided into 3 groups ( n = 18 each): group Ⅰ sham operation (group S); group Ⅱ focal cerebral I/R ( group I/R) and group Ⅲ LXA4 ( group L). Focal cerebral I/R was produced by middle cerebral artery occlusion (MCAO) with a 4-0 nylon thread with rounded tip inserted into right internal jugular vein and threaded cranially in group Ⅱ and Ⅲ . In group Ⅲ LXA4 100 ng was injected into right lateral ventricle of the brain after MCA was successfully occluded. MCAO was maintained for 2 h. The neurological deficit was evaluated and scored (0 = no deficit, 5 = death) at 24 h of reperfusion. 2% Evans blue 4 ml/kg was injected via femoral vein at 1 h before the animals were sacrificed. The animals were killed and their brains were immediately removed for determination of brain water content, Evans blue content and expression of matrix metalloproteinase-9 (MMP-9)in the ischemic cortex. Results The neurologic deficit scores, the brain water and Evans blue content and MMP-9 protein expression in the cortex were significantly higher in I/R group than in S group. The cerebral I/R-induced changes were significantly attenuated in LXA4 group. Conclusion LXA4 can protect blood-brain barrier against cerebral I/R injury by inhibiting MMP-9 protein expression in the brain tissue.

ObjectiveTo evaluate the nephroprotective effects of transplanting metanephric mesenchymal cells (MMCs) into the renal subcaspsule of rats with acute tubular necrosis (ATN) induced by gentamicin. MethodsMMCs were expanded in culture and immunocytochemistry was used to characterize the cells. After gentamicin-induced ATN, fluorescence-labeled cells were transplanted and traced in kidney tissues by fluorescence microscopy. Serum creatinine (Scr) and N-acetyl-b-D-glucosaminidase (NAG) were tested. Kidney pathology was studied by hematoxylin-eosin staining. Apoptosis was examined by the TUNEL assay. Ki-67 and Bcl-2 expression was examined by immunohistochemistry. ResultsMMCs were expanded in culture and the phenotype of the cells was vimentin-positive and keratin-negative. Compared with other ATN groups, in the MMCs-treated group, Scr and NAG clearly decreased[14d Scr: (101.38±20.46) μmol/L vs (248.78±23.15), (252.98±33.52), (229.08±18.18) μmol/L;NAG: (14.83±7.74) U/L vs (33.33±14.88), (29.62±10.54), (30.22±10.94) U/L, P<0.05, respectively];the histopathoiogic lesion scores were lower (P<0.05);the Ki-67 antibody and apoptosis of renal tubular epithelial cells were improved or reduced respectively;the expression of Bcl-2 protein was up-regulated (P<0.05). ConclusionThe subcapsular transplantation of MMCs can ameliorate renal function and repair kidney injury.

Objective To investigate the diagnostic values of procalcitonin (PCT),high sensitive C-reactive protein (hs-CRP),white blood cell (WBC)and percentage of neutrocyte (NEU％)in Gramnegative and Gram-positive bacterial blood stream infection in early stage of sepsis in order to investigate the correlation between PCT and APACHE lⅡ score as well as between PCT and SOFA score,and the prognostic value in assessment of Gram-negative and Gram-positive bacterial blood stream infection.Methods Clinical data of patients admitted to ICU from January 2012 through December 2014 were retrospectively analyzed.A total of 124 sepsis patients with blood stream infection were checked with PCT,hs-CRP,WBC and NEU％ tests,and APACHE Ⅱ score and SOFA score were calculated.The differences in APACHE Ⅱ score and SOFA score were compared between Gram-negative group (n =41) and Gram-positive group (n =83).The correlation between PCT and APACHE Ⅱ score as well as between PCT and SOFA score was analyzed.The differences in diagnostic values of PCT,hs-CRP,WBC and NEU％ between Gram-negative group and Grampositive group were analyzed by using receiver operating characteristic (ROC) curve and it was plotted to assess the prognostic values of PCT,hs-CRP,WBC and NEU％ for septic patients with blood stream infection.Results Compared with Gram-positive group,the levels of PCT [.55.32 (22.01,97.11) vs.2.13 (0.27,5.27)] (P ＜0.01),hs-CRP [105.09 (69.97,186.12) vs.70.54 (42.37,138.63)] (P=0.508),NEU％ [88.30 (75.79,93.52) vs.55.32 (22.01,97.11)] (P=0.302) were higher but WBC was lower [13.59 (10.74,17.58) vs.13.73 (11.32,20.90)] (P=0.058) in Gram-negative group.The ROC curve analysis of PCT showed the area under the curve (AUC) was 0.867 (95％ CI:0.789-0.946).When the optimal cutoff point of PCT was 17.48 ng/mL,the largest Youden's index was found to be 0.661 with 76.9％ sensitivity and 89.2％ specificity.Between two groups,there were significant differences in APACHE Ⅱ score and SOFA score (27.46 ± 9.60 vs.23.67 ± 7.74,P =0.020;8.05 ±3.38 vs.6.59-±3.45,P =0.028).There was significant difference in diagnostic value between PCT and SOFA (r =0.536,P =0.036) in Gram-negative group but no significant difference in Gram-positive group.Conclusions Higher PCT levels are found in Gram-negative group and it can play a role in differntiation between the Gram-negative group and Gram-positive group rather than hs-CRP,WBC and NEU％.PCT can be a better indicator for evaluation of severity of sepsis as well as for prognosis of sepsis patients with Gram-negative bacterium infection.