0.05).But the expression of Caspase-9 in AC group was significantly higher than that in CIN and squamous cell carcinoma group(P

Objective To explore the antitumor effect of interleukin 21(IL-21) after transfected into in HeLa cells in severe combined immunodeficiency(SCID) mice or human peripheral blood lymphocytes-SCID(hu-PBL-SCID) mice.Methods pcDNA4/HisMax-IL-21 was transfected into human cervical cancer cell line HeLa to produce HeLa-IL-21 cells.Totally 48 SCID mice were randomly and equally divided into SCID group(0.2 ml PBS,i.p.) and hu-PBL-SCID group(0.2 ml 2?107/ml hu-PBL,i.p.).Then every group was further divided into 3 subgroups(n=6) to receive a subcutemeous implantation of HeLa,HeLa-vector or HeLa-IL-21 cells,and the rest 6 mice were intraperitoneally injected with anti-asialo GM1 50 mg/animal 1 d before and 4 d after HeLa-IL-21 cells transplantation for NK cell depletion.The growth of the tumor mass was observed.Flow cytometry was used to detect CD8-and DX5-positive cells in hu-PBL from hu-PBL-SCID mice,and spleen NK cells from SCID mice or hu-PBL-SCID mice.The secretion of IFN-? and cytotoxicity of splenic cells from HeLa/HeLa-IL-21-bearing SCID mice or hu-PBL-SCID mice were detected with double sandwich ELISA assay and LDH assay respectively.Results Growth of HeLa-IL-21 tumors was significantly suppressed compared with that of HeLa cells,with a slow development and smaller volume.But this growth suppression was not observed in NK cell depletion groups,with SCID group more severe than hu-PBL-SCID group.hu-PBL-SCID mice bearing HeLa-IL-21 cells(16.55?4.53)had a higher toxicity in spleen cells than those SCID mice bearing HeLa-IL-21 cells(8.32?2.12) or HeLa cells(3.42?1.56)(P

To observe the expression of cyclooxygenase (COX)-1 and COX-2 in brain after spared nerve injury (SNI) and compare the analgesic effects of COX inhibitors with different selectivity. Radioimmunoassay, RT-PCR and Western blotting techniques were used to evaluate the change of brain COX expression at different time points( before SNI, 1 h, 12 h, 1 d, 3 d, 7 d, 14 d, 30 d and 60 d after SNI); By exploring hot plate test, we observed the reacting time of animals after injection of saline, NS-398, SC-560 and indomethacin at different time points. The results showed that: ( 1 ) The expression of brain COX-1 didn't increase significantly until 14 d after SNI, while that of COX-2 increased significantly and rapidly after SNI and reached peak at the time point of 1 d ( all P ＜0.05 ); (2) NS-398 showed significant analgesic effect on neuropathic pain after SNI at the early phase ( P ＜ 0.05 ), but didn't persist for over 30 d; ( 3 ) Indomethacin and SC-560 didn't show significant analgesic effects until 14 d. These results suggest that brain COX-1 is involved in the late phase of neuropathic pain and may play a role in the persistence of pain, while brain COX-2 is involved in the early phase of neuropathic pain and may play a role in the pain origination.

Objective To evaluate the effect of ultra-low dose naloxone on postoperative hyperalgesia caused by large-dose remifentanil.Methods Forty ASA Ⅰ-Ⅲ adult patients,scheduled for gastrointestinal surgery,were randomly assigned into 2 groups (n =20 each):large dose remifentail group (group R) and ultra-low dose naloxone group (group N).Anesthesia was induced with iv injection of remifentanil,propofol and cisatracurium and maintained with inhalation of sevoflurane and infusion of remifentanil.The patients were tracheal intubated and mechanically ventilated.In group R,remifentanil was infused at a rate of 0.25 μg· kg-1 · min-1 starting from the beginning of skin incision.The infusion rate was adjusted according to hemodynamics during operation and subsequently increased/decreased by 0.05 μg· kg-1· min-1 each time.In group N,naloxone was infused at 0.1 μg·kg-1· h-1 while infusing remifentanil,naloxone infusion was stopped at the beginning of peritoneum closure and the other treatments were similar to those previously described in group R.All patients were sent to post-anesthesia care unit after surgery and stayed there for 90 min.Morphine was given when need.The patient-controlled intravenous analgesia was used for postoperative analgesia after leaving post-anesthesia care unit.The first pain time was calculated.The morphine consumption and complications such as nausea,vomiting and pruritus were recorded at 15,30,60 and 90 min and 2,6,24,48 and 72 h after surgery.Results Compared with group R,the morphine consumption was significantly reduced at each time point after surgery,the first pain time was prolonged,and incidence of nausea was decreased (P ＜ 0.05),while no significant change was found in the incidence of vomiting and prutirus in group N (P ＞ 0.05).Conclusion Infusing ultra-low dose naloxone (0.1μg· kg-1 ·h-1) during operation can attenuate postoperative hyperalgesia caused by large-dose remifentanil in patients.

Objective To evaluate the effect of left gastric venous caval shunt in the treatment of portal hypertension. Methods Eight patients suffering from portal hypertension underwent left gastric venouscaval shunt. The graft was of autogenous vein in 5 cases and artificial vein in 3 cases. Results There was no mortality and major complication nor early rebleeding. All patients were followed up from 10 mos to 10 years with an average of 5 years and 2 mos.Postoperatively,5 cases retrieved active living style. Two cases died, and one was lost during the follow-up. Conclusion Left gastric venous caval shunt decreasesthe venous pressure of the portal system within pericardiac and lower esophageal area. The shunt is a safe and effective surgical treatment presenting less alterations to splanchnic hemodynamics and with an additionaladvantage for pericardial devascularization.

Objective To investigate the relationship between high-glucose-induced fibronectin(FN) expression and up-regulation of integrin-linked kinase(ILK) in human kidney tubular epithelial cells (HKC) and kidney of CD-1 mice. Methods Cultured human kidney tubular epithelial cells and streptozotocin (STZ)-indueed diabetic model of CD-1 mice were enrolled in this study.Western blot was used to detect the expression of FN and ILK.The kinase dead ILK plasmid (pCMV-kdlLK) were transferred to HKC. Results Four weeks after injection of STZ,CD-1 mice had higher blood glucose level as compared to the control [(20.3±2.7) mmol/L vs (6.1±1.4) mmol/L,P＜0.01].Meanwhile,expression of FN and ILK was significantly increased in diabetic mice as compared to the control (P＜0.01).There was positive correlation between the expression of FN and ILK (r=0.899,P＜0.01).High-glucose could up-regulate FN and ILK expression in cultured HKC in a time- and dose-dependent manner.Blockage of ILK activation by pCMV-kdILK abrogated high-glucose-incuced FN expression in HKC. Conclusions Highglucose can induce FN expression through up-regulating ILK expression.Blockage of ILK activation abrogates this effect.

Objective To investigate the effect of high glucose on renal tubular epithelial-mesenchymal transition,and to analyze the relationship between high glucose and transforming growth factor β1(TGF-β1)and the mechanism of renal interstitial fibrosis. Methods HKC and Smad7-overexpression HKC cells were grown in DMEM/F12 medium containing 5%～10%newborn calf serum.They were cultured for 16 h in free serum medium after 80%cells were adhered onto the surface of the flask.Afterwards,they were stimulated by high glucose(glucose concentration:25 mmol/L and 50 mmol/L).The expression of α-SMA,E-cadherin and fibronectin was detected by Western blot while the supernatant level of TGF-β1 was detected by ELISA.Cell motility and migration was evaluated using Boyden chamber motogenicity assay. Results In HKC induced by high glucose,the expression of α-SMA and fibronectin protein was highly upregulated while the expression of E-cadhefin protein was down-regulated.The expression of TGF-β1was up-regulated in a dose-dependent manner.These above-mentioned effects could be obviously inhibited by anti-TGF-β1 antibody.The protein expression of α-SMA,fibronectin and E-cadherin had no obvious change in Smad7-overexpression HKC induced by high glucose.HKC exhibited enhanced motility and invasive capacity in high glucose groups,compared to that in control group.Migrated cell counting was(12.4±3.7)and(18.6±4.4)cell/HP in 25 and 50 mmol/L glucose groups respectively. Conclusion High glucose may induce renal tubular epithelialmesenchymal transition through TGF-β1 pathway,which can be inhibited by blocking the Smad signal pathway.

AIM: To compare the expression of three cyclooxygenase (COX) isoforms in the process of inflammatory pain and evaluate the analgesic effects of different protocols about usage of COX inhibitors on inflammatory pain. METHODS: Formalin was injected subplantarly to mice to induce inflammatory pain. The expression of COX-1, COX-2 and COX-3 was evaluated by radioimmunoassay and RT-PCR, respectively. For the analgesic effect assay, animals were divided into 5 groups including control, SC, NS, IN and NS + SC group. The former 4 spectively. In the NS + SC group, animals received NS398 during the first 1 month and SC-560 during the second month in the NS + SC group. RESULTS: The expression of COX-1 was higher at the late phase while that of COX-2 was higher at the early phase of inflammatory pain. The expression of COX-3 did not significantly change in the process of inflammatory pain. Additionally,behavioral assessment showed that using COX-2 inhibitors at the early phase followed by COX-1 inhibitors at the late phase could get better analgesic effect on inflammatory pain compared with single using COX-1 selective or COX-2 selective inhibitors. CONCLUSION: In brain, the expression of COX-2 increases rapidly in the inflammatory pain process while COX-1 expression does not increase till the late phase. Brain COX-3 is poorly involved in the inflammatory process. Combined use of COX-1 and COX-2 selective inhibitors may be a better protocol in inflammatory pain treatment.

OBJECTIVE Through investigation of the changes in biological characteristics of an E scherichia coli bacteriophage with broad host range,to study the mechanism of phage-host specificity and the bactericidal efficacy on sewage water samples.METHODS In this study,one strain of E.coli bacteriophages with broad host range was screened from natural environment using the E.coli 285.Then,in order to analyze the difference of biologic effects before and after the host range changed,we made a comparison of biological properties between the phage with broad host range and the phage f2.RESULTS The ultrastructure under electron microscope showed that the two phages were round-shaped particles,but phage XY′s diameter between 20-90 nm,short-tailed and occasionally visible;the anti-serum K value of phages f2 and XY was 30.1 and 41.5,respectively;whereas,between phages XY and f2,there was a low correlativity(K value

Objective To investigate the growth inhibition and DNA damage of energized fusion protein anti-CD20(Fab)-LDM on B JAB cells in vitro.Methods The binding activity of fusion protein anti-CD20 (Fab)-LDP to B JAB cells was studied by flow cytometry and confocal laser scanning microscopy.MTT assay was used to study the energized fusion protein anti-CD20(Fab)-LDM on cell growth of B JAB cells.Comet assay was employed to detect DNA damage in B JAB cells.The cell growth cycle of BJAB was analyzed by FACS.Results The recombinant fusion protein anti-CD20 (Fab)-LDP possessed an significant target affinity towarded BJAB cells.The energized fusion protein anti-CD20(Fab)-LDM showed obvious inhibition on proliferation,as well as induced potent DNA damage in B JAB cells in vitro compared with lidamycin.B JAB cells treated with energized fusion protein anti-CD20 (Fab)-LDM showed S phase cell cycle.Conclusion The energized fusion protein anti-CD20 (Fab)-LDM could target binding to BJAB cells and significantly inhibit the proliferation of B JAB cells by inducing DNA damage and S phase arrest.