The in vitro tumoricidal activity of peritoneal macrophages harvested from C57BL mice and activiated by li posomes containing water-soluble muramyl dipeptides (MDP) was measured by ~(125)I.UdR release assay. The results demonstrated that liposomes containing MDP could effectively activiate macrophages to become tumoricidal. The percentage of pu(?)monarymetastasis and the average metastatic pulmonary colonies of mice with experimental tumor metastasis decreased after intravenous injection of MDP encapsulated in liposomes. These results suggested that systemic administration liposomes containing MDP whichinduced macrophage activiation might provide a useful immunot heray for tumor, especial1y for pulmonary mini metastasis.

Objective To investigate the changes of plasma fibrinolytic activity and D dimer level in the primary hypertension of the elderly.Methods ELISA was applied to measure blood and urine D dimer.Chromophore substrate method was used to detect PL,Plg,t PA and PAI 1.Results The levels of PAI 1,D dimer and Plg were obviously higher in the elderly primary hypertension and hypertentive kidney disease than those in the control group,while t PA and PL activity were lower than in the control group.Conclusion The changes of PLA and D dimer play an important role in hypertension and hypertentive renal disease of the elderly,on which the early diagnosis can be based.

Objective To investigate the effect of triptolide on asthmatic airway remodeling and signal transducer and activator of transcription 6 (STAT6), acid neutrophil chemokines (eotaxin) impact. Methods The total of 30 mice with ovalbumin (OVA) model of asthma were randomly divided into three groups, control group, asthma group and triptolide group. After 24 hours of the last shot, lung tissue was stained Bronchial inflammatory cell infiltration was determined by using semi-quantitative method and calculate the proportion of goblet cells in airway epithelial cells. Hydroxyproline was determined by McMillan airway mucus score. The mRNA level and protein level of STAT6 and eotaxin in airway epithelium were determined by RT-PCR and immunohistochemistry. Results Compared with asthma group, peribronchial inflammatory cells infiltration of triptolide group were reduced, which mucus index is (1.31 ± 0.23) and hydroxyproline is (284 ± 13) μmg/100 mg. it had a significant in asthma group (P < 0.05). Besides, the protein level and mRNA level of STAT6 and eotaxin were significantly decreased (P < 0.05). Moreover, it was a positive correlation between STAT6 and eotaxin level in airway epithelial (r = 0.668, P < 0.05). Conclusion Triptolide can inhibit airway remodeling and might through the down regulation of STAT6 and eotaxin expression.

Aim To study the sensitivity of human gastric cancer BGC-823 cells to paclitaxel after trans-fection of SHIP2 ( The SH2 domain containing inositol 5-phosphatase 2 ) cDNA. Methods Apoptotic cells were determined by the propidium iodide method using flow cytometry. The levels of protein and mRNA ex-pression were measured by Western blot analysis and qRT-PCR, respectively. pCMV6-SHIP2 plasmid and empty vector were transiently transfected into BGC-823 cells, respectively. Stable cell lines were established after infecting BGC-823 cell with GV112-Puromycin and GV112-Puromycin-Bim( Bcl-2 interacting mediator of cell death) lentivirus particles. pCMV6-SHIP2 plas-mid was transiently transfected into the stable cell lines. Results BGC-823 cells were relatively insensi-tive to paclitaxel compared with SGC-7901 cells. The apoptotic rate was only (25. 6 ± 1. 6)% after the treat-ment with 0 . 3 μmol · L-1 paclitaxel for 48 h in BGC-823 cells. The expression levels of Bim protein and mRNA in BGC-823 cells treated with paclitaxel at dif-ferent time points were not significantly changed. The expression of Bim protein was increased after transfec-tion of pCMV6-SHIP2 plasmid, and the apoptotic rate was up to ( 50. 8 ± 0 . 9 )% in BGC-823 cells treated with paclitaxel for 48h. The expression of Bim protein was significantly inhibited after infecting with GV112-Puromycin-Bim lentivirus particles. The apoptotic rate of infected BGC-823 cells was only ( 27. 6 ± 1. 6 )%after treatment upon paclitaxel for 48h. Conclusion Overexpression of exogenous SHIP2 can increase the expression of Bim, induce apoptosis and enhance sen-sitivity of BGC-823 cells to paclitaxel.

Objective To investigate the mechanism of apoptosis induced by UMI-77 , a novel selective inhibitor of Mcl-1, in gastric cancer MGC-803 cells. Methods MGC-803 cells were treated with UMI-77 in different concen-trations for 24 h, apoptotic rates were determined by Annexin V/PI method using flow cytometry. Mitochondrial membrane potential was determined by JC-1 staining on a flow cytometer. The activation of Caspase-9, Caspase-3 and cleavage of PARP were measured by Western blot analysis. The protein level of Bcl-2, Bcl-XL and Mcl-1 was monitored by Western blot as well. Chemically synthesized Mcl-1 siRNA was transfected into MGC-803 cells using Lipofectamine 2000 Reagent. The efficacy of gene silencing was confirmed by Western blot analysis, and opoptotic rates before and after transfection was measured by flow cytometry using Annexin V/PI staining. Results UMI-77 was effective in induction of apoptosis in gastric cancer MGC-803 cells, apoptotic rates were increased in a dose-de-pendent manner. Mitochondrial membrane potential was collapsed after UMI-77 treatment. Activation of Caspase-9, Caspase-3 and cleavage of PARP occurred at 24 h (P<0. 05). The expression level of Bcl-2 and Bcl-XL were not altered after exposure to UMI-77 , while Mcl-1 was down-regulated after 12 h ( P<0. 05 ) . Transfection with Mcl-1 siRNA successfully decreased the expression level of Mcl-1 in MGC-803 cells ( P<0. 05 ) and blocked apoptosis induced by UMI-77 ( P<0. 05 ) . Conclusion UMI-77 induces apoptosis through activation of the intrinsic path-way in gastric cancer MGC-803 cells, and knocking down Mcl-1 expression abrogates apoptosis by UMI-77.

Objective To investigate the effects of Arctigenin ( ATG ) on concanavalin ( ConA )-stimulated cell proliferation and cytokine secretion in mouse spleen cells, and its possible mechanism. Methods The toxicity of ATG on mouse spleen cells was determined by MTT assay. The inhibition of proliferation was investigated by tritiat-ed thymidine incorporation method. Secreted cytokines (IFN-γand IL-2) were analyzed by ELISA. The associated proteins and phosphorylation levels of mTOR pathway ( mTOR/P70 S6 K/Akt/AMPK/Raptor ) were detected by Western blot. Results ATG had no significant toxicity to mouse spleen cells. ATG significantly inhibited mouse primary spleen cells proliferation induced by ConA. ATG suppressed IL-2 and IFN-γ production of mouse spleen cells in a concentration-dependent manner. ATG remarkably suppressed the phosphorylation of mTOR and P70S6K, and enhanced the phosphorylation of upstream AMPK and Raptor, while the phosphorylation of Akt did not change significantly. Conclusion ATG markedly suppresses the proliferation of mouse spleen stimulated by ConA cells and secretion of IFN-γand IL-2 , which may be correlated to the abilities of enhancing the phosphoryla-tion of AMPK and Raptor, inhibiting the phosphorylation of mTOR and P70S6K.

Objective To investigate the role of anti apoptosis gene bcl 2 in the survival of cultured uveal melanoma UM cells. Methods Antisense oligodeoxynucleotides (AS ODN) bcl 2 were delivered with cationic lipid to primary cultured UM cells. The inhibitory effect of AS ODN bcl 2 on proliferation of UM cells was examined by 3, 4,5 Dimethyliazol 2,5 diphenyl tetrazolium bromide (MTT) method. Using DNA ladder to determine if the UM cells had been apoptotic. Bcl 2 expression was detected by RT PCR and Westernblot technics. Results The effect of AS ODN bcl 2 in inhibiting the proliferation of cultured UM cells had opposite relation to dosage. It down regulated the mRNA and protein level of bcl 2 gene, and the sense ODN didn′t have this effect. Conclusion AS ODN bcl 2 can down regulate bcl 2 expression, inhibits UM cells proliferation and induces apoptosis.

Aim To investigate the potential involve-ment of caspase-4 in TRAIL ( tumor necrosis factor-re-lated apoptosis-inducing ligand )-induced apoptosis in gastric cancer cells. Methods The effect of treatment with TRAIL /z-LEVD-fmk alone or in combination for 24 h on the apoptotic rate of gastric cancer cells was detected by FCM ( flow cytometry) using propidium io-dide DNA staining. Chemically synthesized three siR-NAs targeting caspase-4 gene were transfected into gas-tric cancer cells by Lipofectamine 2000 Reagent. The efficacy of gene silencing was confirmed by Western blot analysis . The apoptotic rates of gastric cancer cells to TRAIL before and after transfection with caspase-4 siRNA were observed by FCM. The expression level of GRP78 (78-ku glucose-regulated protein) protein was examined by Western blot, the classic endoplasmic re-ticulum stress inducer tunicamycin ( TM) was used as a control. The expression levels of caspase-4 and caspase-3 after TRAIL treatment were also measured by Western blot. Results z-LEVD-fmk decreased TRAIL-induced apoptosis of gastric cancer cells signifi-cantly(P<0. 05). As compared with negative control, the expression level of caspase-4 protein was reduced after transfection, and the apoptotic rate was also de-creased ( P <0. 05 ) . While TM induced marked up-regulation of GRP78 , treatment with TRAIL resulted in, albeit to a lesser extent, increases in GRP78, in-dicative of induction of ER stress by TRAIL. Activa-tion of caspase-4 and caspase-3 occurred early after TRAIL treatment. Conclusion Activation of caspase-4 contributes to TRAIL-induced apoptosis and is asso-ciated with induction of ER stress by TRAIL in gastric cancer cells.

Objective To evaluate the patient-controlled paravertebral block (PCPB) in optimizing the cellular immune function when used after radical resection of pulmonary carcinoma performed via video-assisted thoracoscope in patients.Methods Forty-one ASA physical status Ⅰ or Ⅱ patients of both sexes,aged 50-64 yr,with body mass index of 20-25 kg/m2,of TNM staging Ⅰ or Ⅱ,undergoing radical resection of pulmonary carcinoma performed via video-assisted thoracoscope,were randomly divided into 2 groups using a random number table:PCIA group (n =21) and PCPB group (n =20).PCIA solution contained sufentanil 2 μg/kg in 100 ml of normal saline.The PCIA pump was set up to deliver a 2 ml bolus dose with a 15-min lockout interval and background infusion at 2 ml/h.In PCPB group,the patients received paravertebral injection of 0.2％ ropivocaine 5 ml at T5 level on the affected side under ultrasound guidance at the end of operation,and then received PCPB.PCPB solution contained 0.75％ ropivacaine 67 ml in 250 ml of normal saline,and the pump was set up to deliver a 5 ml bolus dose,with a 15-min lockout interval and background infusion at 5 ml/h.VAS score was maintained ≤ 3,and analgesia lasted until 50 h after operation.Before induction of anesthesia (baseline),at end of operation,and at 1,3 and 5 days after operation,peripheral venous blood samples were collected to determine the levels of regulatory T cells,natural killer cells and natural killer T cells (by flow cytometry) and plasma concentrations of interleukin-10 and transforming growth factor-β (by ELISA).Results Compared with group PCIA,the level of regulatory T cells was significantly decreased,the levels of natural killer cells and natural killer T cells were increased,and the plasma concentrations of interleukin-10 and transforming growth factor-β were decreased at 1 and 3 days after operation,and no significant change was found in the rate of cellular immune function decline after operation in group PCPB.Conclusion PCPB provides no significant difference clinically in optimizing the cellular immune function when used after radical resection of pulmonary carcinoma performed via video-assisted thoracoscope in the patients.

Objective To obtain genes encoding the novel molecules for diagnosis of schistosomiasis.Methods Juvenile S.japonicum cDNA library was immunoscreened to obtain positive clones.By DNA sequencing and sequence analysis,the target gene was amplified by PCR and subcloned into prokaryotic plasmid pET28a.The recombinant plasmid was transformed into E.coli BL21 followed by expression of the protein induced by IPTG.The protein was identified by Western blotting.Results 34 positive clones were obtained,24 of which were chosen to be sequenced,13 of which were Sj22600 gene.The protein were recognized with sera of infected rabbits and patients with acute and chronic schistosomiasis by Western blotting.Conclusion The gene coding for Sj22600 membrane protein was screened with high frequency in the cDNA library.The E.coli BL21 transformed with the recombinant plasmid can express the fusion protein,which shows immunoactivity.