Objective To evaluate the results of endovascular intervention for infrapopliteal arterial occlusion in 40 patients. Methods There were 41 affected limbs in these 40 patients receiving 44 times of endovascular intervention for infrapopliteal arterial occlusion during Nov. 2006 and Dec. 2007. The average age was 76±6. The ABI(ankle brachial index)before intervention was 0.39±0.20 in anterior tibial artery and 0.39±0.23 in posterior tibial artery. CLI (critical limb ischemia) was 80.49% (33/41). Results The after intervention ABI increased by 0.43±0.22 (P<0.01) in anterior tibial artery and 0.43±0.25(P<0.01)in posterior tibial artery. 35 patients (36 limbs) were followed-up for (6±3) months. The limbs of Fontaine Ⅰ and Fontaine Ⅱ A were 28 (77.78%), CLI decreased to 19.44% (7/36) (P<0.01). At follow-up the ABI in anterior tibial artery was 0.63±0.22 and 0.56±0.22 in posterior tibial artery. The difference were all significant when compared with that before intervention and after intervention. The perioperative amputation rate was 0. The perioperative mortality rate was 2.5%. The total mortality rate was 15%. The limb salvage rate were 100%. Conclusion The effect of endovascular intervention for infrapopliteal arterial occlusion is satisfactory.

Objective To study the resistances of CDl33 + subset purified from gastric cancer cell line to chemotherapy drugs and the mechanism of this resistance regarding to the mRNA expressions of both Bcl-2 and BAX in relation to the relative apoptotic genes.Methods CD133 + subset and CD133-subset were purified from KATOⅢ cell linc by magnetic activated cell sorting.The proliferating ability of these two subsets resistantnt to 5-FU,Cisplatin(DDP,PDD) and Etoposide was checked and compared by CCK-8 test.The apoptotic changes of these two subsets regarding to the expression of mRNA of both Bcl-2 and BAX were also analized by RT-PCR.Results In CD133 + subset,the contant percentage of CD133 + expression rate was 90％ via analysis of flow cytometye.Twelve hours after treatment of5-FU,DDP and VP-16,the cells in both CD133 + subgroup and CD133-subgroup would gradually start to change in apoptotic morphology.The growth inhibiting rate by CCK-8 measurement for 5-FU,DDP and VP-16 groups in CD133 + subgroup was significantly lower than that in CD133-subgroup.The data under different treatment respectively was,5-FU:(30.56 ± 1.99) ％-(88.60 ± 1.95) ％ vs (32.81 ± 2.67) ％-(95.73±2.12)％,P=0.045,cisplatin:(45.89 ±3.64)％-(81.20 ± 1.18)％ vs (50.21 ±3.22)％-(90.46±1.89)％,P=0.043,VP-16:(37.21 ±3.80)％-(78.49 ±3.22)％ vs (35.55 ±3.23)％-(89.32 ±-3.54) ％,P =0.048).After treatment of these three kind of anti-tumour drugs,the expression level of Bcl-2 mR-NA decreased significantly and the expression level of BAX mRNA increased significantly in both CD133 + subgroup and CD133-subgroup.However,these changing ranges of Bcl-2 mRNA and BAX mRNA were more obvious in CD133 + subgroup in comparison with those in CD133-subgroup.Conclusions In some degree,resistent potentiality of CD133 + cells to 5-FU,DDP and VP-16 has been identified,which may probably be due to the up-regulation of the expression of BAX and down-regulation of the expression of Bcl-2.

The paper is to report the establishment of a method for the determination of multi-residue organochlorine and pyrethroid pesticides in traditional Chinese medicines (TCMs). Fifty-six pesticides were extracted by high-speed homogenization, and then purified through gel permeation chromatography (GPC) and solid phase extraction (SPE) cartridges. The residues were simultaneously identified and quantified by GC-ECD equipped with dual tower, dual column and two micro-ECD detectors. The analytical performance was demonstrated by the analysis of 3 TCMs samples' extracts, spiked at three concentration levels for each pesticide. In general, the recoveries ranging from 70% to 110%, with relative standard deviations (RSDs) better than 15%, were obtained. The limit of detection (LOD) for most of the targeted pesticides tested was below 0.01 mg kg(-1). The method had good extraction efficiency, purification effect and good reproducibility, which could be applied to the determination of organochlorine and pyrethroid pesticide residues in the routine analysis of TCMs.

Objective To investigate the clinicopathologic characters and prognostic value of epithelial mesenchymal transition (EMT) related proteins Snail and chemokine receptors 4(CXCR4) in gastric cancer.Methods The expressions of Snail and CXCR4 in the gastric tissues of 50 patients with gastric cancer were detected by Immunohistochemistry.The relation between the expressions of Snail and CXCR4 and the clinicopathologic characters were analyzed.The relative relationship between Snail expression and CXCR4 expression were identified by Spearman method.The correlation between the expressions of Snail and CXCR4 with the survival was analyzed by Kaplan-Meier method.Results The expression of Snail and CXCR4 expression were positive in 31 of 50 cases (62％) and 29 of 50 cases (58％) respectively.The expression levels of Snail and CXCR4 in the gastric cancer patients with diameter more than 5 cm,worse tissue differentiation,vascular invasion,lymphatic vessel invasion,lymph nodes metastasis,and T3 + T4 staging were significantly higher than those in the patients with diameter less than 5 cm,without vascular invasion,lymphatic vessel invasion,lymph nodes metastasis,and T1 + T2 staging (P ＜ 0.05).The Expression of Snail and CXCR4 was positively Correlated(r =0.330,P =0.014).The co-expression of Snail and CXCR4 was significantly associated with poorer prognosis and was applied as an independent prognostic factor for gastric cancer (P =0.001).Conclusions The co-expression of Snail and CXCR4 was the independent prognostic factor for gastric cancer.The combining effect of EMT and CXCR4 may promote the progressions and metastasis of gastric cancer.Therefore,it may be of an effective way for the metastasis of gastric cancer to adjust these targets of the Snail and CXCR4.

ObjectiveTo investigate the expression of CXCR4 and CD133mRNA in primary lesion of primary gastric adenocarcinoma and the relation with clinicopathological features,and to explore the correlation of CXCR4 and CD133.MethodsThe primary lesion of primary gastric adenocarcinoma and normal tissues adjacent to gastric cancer were obtained from 50 patients.The diction of CXCR4 and CD133 protein expression was detected by the immunohistochemical staining,and the relative gray scale values of CXCR4 and CD133mRNA by semi-quantitative RT-PCR (Fisher' s exact probability method).Their relationship with clinicopathological features was also investigated ( Spearman relation analysis).ResultsThe positive rates of CXCR4 and CD133 protein in gastric cancers were 76.0％ and 66.0％ respectively,which were significantly higher than that in normal tissues adjacent to gastric cancer ( 16.0％ and 10％ ; P =0.000,P =0.000).The increment of relative gray scale values of CXCR4mRNA was associated with the larger tumor diameter,the later TNM stage and the occurrence of lymphatic metastasis( P ＜ 0.05 ).And the larger diameter of tumor,the later TNM stage were associated with the higher relative gray scale values of CD133mRNA (P ＜0.05).The levels of the relative gray scale values of CXCR4 mRNA and CD133mRNA were positively related(r =0.453,P ＜ 0.01 ).ConclusionsThe higher expression of CXCR4 and CD133mRNA correlateswith tumour diameter,TNM stage and lymphatic vessel invasion. The relative gray scale values of CD133mRNA increase with the increment of the relative gray scale values of CXCR4.

AIM: The multi-residues method was used to determine organophosphorus pesticides in traditional Chinese herbal medicines (TCHMs).METHODS: Fifty three pesticides were extracted by high-speed homogenization,and then cleaned sequentially by C_(18) and Carb/NH2 solid phase extraction (SPE) cartridges.The residues were simultaneously identified and quantified by GC-FPD equipped with dual tower,dual column and two FPD detectors.RESULTS: The analytical performance was demonstrated by the analysis of 6 TCHMs samples extracts,spiked at three concentration levels for each pesticide.In general,the recoveries ranging from 70% to 120%,with relative standard deviations (RSDs) less than 15%,were obtained.The limit of detection (LOD) for most of the targeted pesticides tested was below 0.01 mg/kg.CONCLUSION: The method has good extraction efficiency,purification effect and good reproducibility,which can be applied to the determination of organophosphorus pesticide residues in the routine analysis of TCHMs.

AIM:To study a method for the determination of 13 N-methylcarbamate peticides in traditional Chinese herbal medicines(TCHMs).METHODS:Thirteen pesticides were extracted by acetonitrile,and purified through solid phase extraction(SPE)cartridges,then detected by HPLC with post-column derivatization and Fluorescense Detector excited wavelength was 330 nm,emissive wavelength was 465 nm.RESULTS:The analytical performance was demonstrated by the analysis of 6 TCHMs samples' extracts,spiked at three concentration levels for each pesticide.In general,the recoveries ranging from 74.1 to 108.8%,with relative standard deviations(RSDs)better than 15%,were obtained.The limit of quantification(LOD)of 13 pesticides from 0.000 3 to 0.06 mg/kg.CONCLUSION:The method has good extraction efficiency and purification effect,which could be standard of N-methylcarbamate peticides residues in the routine analysis of TCMs.

Objective To investigate the potential effect of glucagon-like peptide-1 (GLP-1) analogue exendin-4 (Ex-4) on immune function of T lymphocytes via neuroendocrine modulation mechanism in mice following severe burns.Methods Male BALB/C mice were randomly (ramdam number) divided into thermal injury group (n =50) and sham-thermal group (n =30).The thermal injury model was made by exposing the back skin of 15％ total body surface area (TBSA) to 95 ℃ water for 7 seconds,while in sham-thermal model the mice were immersed in 37 ℃ water instead.The expression of GLP-1 receptor (GLP-1 R) was determined in sorted CM + T cells from normal mice by immunofluorescence method.In ex vivo experiment,the mice were sacrificed at 24 h post-bum,then the mononuclear cells (MNC) from spleen were separated from both groups and cultured in RMPI 1640 with 10％ FCS (fetal calf serum) in presence of ConA (concanavalin A,5 μg/mL).Cells were pretreated with catecholamine receptor antagonist propranolol (prop) for 1 hour,followed by consecutive dose of Ex-4 for another 48 h.In in vivo experiment,prop (30 mg/kg) was i.p.injected 30 minutes before thermal injury,then Ex-4 (2.4 nmol/kg) was injected i.p.immediately after scalding.Mice were sacrificed at 6 h and 24 h after thermal injury,then the serum and the spleens were collected.Results GLP-1R was expressed on splenic CD4 + T cells from normal micc.Ex-4 exerted no marked effect on the functions of T cells in terms of proliferation and IL-2 secretion at all doses examined ex vivo,which was not affected by pretreatment with prop.In vivo,T cell functions were suppressed by Ex-4 in thermal mice (P ＜ 0.05),but was restored by pretreatment with prop.Regardless of ex vivo or in vivo,Ex-4 could induce T cells switched to Th2 response (P ＜ 0.05).Moreover,the Th2 switch by Ex-4 was greatly potentiated by prop intervention in thermal mice in vivo other than ex vivo.Norepinephrine level was increased and epinephrine was decreased by Ex-4 in thermal mice.Both norepinephrine and epinephrine levels were obviously enhanced by pretreatment with prop.Conclusions Ex-4 can inhibit the proliferation and IL-2 secretion of splenic T lymphocytes through the sympathetic nervous system,however,it might induce Th2 switch from Th cells by acting directly on GLP-1R.

Objective To compare the inhibition effects of three synthesized fragments used in small interfering RNA(siRNA) against CD133 gene in KATO-Ⅲ gastric cancer cells,and to study effects of suppressed CD133 on the proliferating ability of intervened cells.Methods Three fragments of siRNA were designed and synthesized targeted at the mRNA of CD133.Cell fluorescence counting under confocal laser scanning microscope was used to determine the transfection efficiency after transfection with the CD133FITC-siRNA.The knock-down effect of the CD133 gene was detected by RT-PCR and Western blotting.CCK-8 (cell counting kit-8 assay) was performed to measure the variation of the cell proliferative viability after the above-mentioned treatment.Results The transfection efficiency of siRNA was (85 ± 8) ％ in KATO Ⅲ Gastric cacer cell.All these three fragments of CD133 siRNA effectively inhibited the expression of CD133 gene,the inhibition rate being (11 ± 2) ％,(19 ± 2) ％,(24 ± 3) ％respectively.Compared with the control group,the cell proliferation viability was restrained (42 ± 4)％ in CD133siRNA-3 group (P ＜0.05).Conclusions CD133siRNAs were successfully transfected into KATO Ⅲ Gastric cacer cells and repressed the expression of CD133.Meanwhile,the CD133siRNA fragment 3 was screened from three CD133 siRNA,which has the best inhibition effect.The results provide preliminary evidence for the intereference of CD133+ gastric cancer cells subsequently.

Objective To investigate the biological effect of epithelial-to-mesenchymal transition (EMT) under the treatment of transforming growth factor (TGF)-β1 on the human gastric cancer cell line SGC7901 in vitro,and to observe whether the TGF-β1 can generate the tumor initiating cells ability in SGC7901 or not.Methods SGC7901 cells were cultured with TGF-β1.The morphological change was observed.The effect on proliferation of SGC7901 cells was detected by CCK-8.The invasion assay was used to investigate the motility and the invasion ability of SGC7901 cells.Immuofluorescence was used to detect the expression of E-cadherin and N-cadherin.The mRNA and protein's expression levels of EMT-related factors and CD44 were analyzed by RT-PCR and Western blotting respectively.Results TGF-β1 induced morphological alterations from epithelial to mesenchymal cells.The proliferation of SGC7901 cells was inhibited,and the ability of motility and invasion of SGC7901 cells were greatly enhanced after being treated with TGF-β1.RT-PCR and Western blotting showed that the expression of Snail (P ＜ 0.05),N-cadherin (P ＜ 0.05) and CD44 (P ＜ 0.05) were significantly increased while the expression of E-cadherin was decreased (P ＜ 0.05).Conclusions TGF-β1 can generate the EMT.The CD44 expression was up-regulated.TGF-β1 can inhibit the proliferation and promote the motility and invasion ability of SGC7901 cells.