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

0.05).But the expression of Caspase-9 in AC group was significantly higher than that in CIN and squamous cell carcinoma group(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 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 evaluate the applicability and security of transpancreatic precut sphincterotomy vs double guidewire technique for cannulation in difficult bile duct cannulation in endoscopic retrograde eholangiopancreatography (ERCP). Methods Retrospective analysis of 158 cases difficult bile duct cannulation in ERCP from January 2012 to January 2014, according to the intubation tube method, we divided all the cases into 3 groups, transpancreatic precut sphincterotomy group (group A); double guide wire technique group (group B); single guide wire technique group (group C). Then compare the intubation success rate and the incidence of complications among the 3 groups. Results 54 of 58 patients in group A intubation successful, the success rate is 93.1%, 50 of 56 patients in group B intubation successful, the success rate is 89.3%, 26 of 44 patients in group C intubation successful, the success rate is 59.1%, there was no significant difference between group A and B(P > 0.05), group A and group C, group B and C have significant difference (P < 0.05). In group A, 4 cases were complicated with acute pancreatitis, hemorrhage in 6 cases, infection in 2 cases, the complication rate is 20.7%; In group B, 5 cases were complicated with postoperative pancreatitis, 4 cases of infection, incidence of complications is 16.1%; 7 patients were complicated with pancreatitis in group C, hemorrhage in 2 cases, infection in 4 cases, complication rates is 29.5%, 3 groups were no perforation occurred.The complication rate of group B is lower than in group A, but no significant difference (P > 0.05), group A and group C, B and C complication rates had significant difference (P < 0.05). Conclusions When selective bile duct intubation is difficulty and guide wire thread into the pancreatic duct, continue to single guide wire have low intubation success rate and higher incidence of complications,transpancreatic precut sphincterotomy and double guide wire technique can effectively improve the success rate of intubation, and complication rates are relatively low, no significant difference between the two.

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.

BACKGROUND:An abroad study repoRed the distribution and expression of augmenter of liver regeneration(ALR)in the central nervous system.There are few literatures on how to prepare and evaluate ALR protein polyclonal antibody in recombinant rats,and how to construct prokaryotic expression vector.There are no repots concerning ALR in the central nervous system in China.OBJECTIVE:TO express ALR fusion protein in E coli BL21 and prepare and identify polyclonal antibody.METHODS:RNA was extracted from the hippocampus of Sprague Dawley rats.The prokaryotic expression plasmid pET28a-ALR was constructed and the positive recombinant plasmid was transformed into BL21.Protein ALR was expressed by inducing transformed BL21 with Isopropyl-β-D-thiogalactopyranoside(IPTG)and purified by Ni~(2+)affinity chromatography column after immune the rabbit for 4 times.the serum of rabbits was extracted from hear as polyclonal antibody.The titer and specificity of the rabbit's antiserum was respectively measured by ELISA and Western blotting The following parameters were measured:construction of prokaryotic expression plasmid pET26a-ALR;pET28a-ALR recombinant enzyme digestion evaluation;results of ELISA and Western-blotting.RESULTS AND CONCLUSION:Expecting bands were obtained by double enzyme digestion electrophoresis,respectively 5.3 kb and 0.4 kb.Nucleotide sequence analysis verified that prokaryotic expression vector pET28a-ALR was successfully constructed.The 19 ku fusion protein was successfuIly expressed.The titer of the antiserum measured by ELlSA could achieve 1:2 000 This indicated that antibody and purified recombinant ALR had a good reaction.and high titer.could meet the experimental require.Western blotting analysis proved that the antibody could identify the prokaryotic expression product of ALR.Prokaryotic expression system expressed ALR fusion protein,prepared and purified polyclonal antibody of ALR protein,and could meet the experimental require of ALR immunoblotting.