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.

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.

Objective To analyze the endoscopic features of patients with the lower esophageal sphincter (LES) and crural diaphragm (CD) separation and explore its significance in hiatus hernia (HH) diagnosis.Methods From January 2011 to June 2012,the data of patients underwent esophagus high resolution manometry (HRM) examination were retrospectively analyzed and of which the patients with LES-CD separation were selected.Patients who received digestive tracts operation or lack of complete endoscopic date were excluded.A total of 93 cases were enrolled for analysis.According to Chicago Criteria,the cases were divided into three groups.There were 21 cases in type Ⅰ group (LES-CD separation＜1 cm),37 cases in type Ⅱ group (LES-CD separation≥l cm and ＜2 cm),35 cases in type Ⅲ group (LES-CD separation≥2 cm).The differences among groups in endoscopic features were analyzed.HRM were taken as golden standard,the sensitivity and specificity of endoscopy in HH diagnosis were analyzed.Measurement data were expressed as x ± s,one-way ANOVA was performed for multi-groups comparisons,LSD method or Dunnett T3 test for pairwise comparisons and chi-square test or Fisher's exact test for two groups' comparisons.Results There were no significant differences in the distance between esophagus gastric junction (EGJ) and foreteeth of the three groups (P＞0.05).Distances between EGJ and diaphragmatic HH pressure impression in type Ⅲ group,type Ⅱ group and type Ⅰ group were (3.57±0.78) cm,(1.89±0.81) cm and (1.14±0.67) cm,respectively and the differences were significant (LSDt=9.26,11.44,3.57; all P＜0.05).The percentage of continuously-open cardia and showing pressure impression of the gastric pouch of type Ⅲ group was higher than that of type Ⅱ group and the percentage of continuously-open cardia and showing pressure impression of gastric poach of type Ⅱ group was higher than that of type Ⅰ group (80.0％,40.5％ and 4.8％,x2 =11.64,29.76 and 8.59; 91.4％,27.0％ and 4.8％,x2 =30.69,40.73 and 4.32' all P＜0.05).The occurrence rates of the His angle blunting and sac of type Ⅲ group were higher than those of typeⅡ group and type Ⅰ group (74.3％ and 77.1％,24.3％ and 24.3％,4.8％ and 4.8％,x2 =17.97 and 25.41,both P＜0.05),there was no significant difference between type Ⅱ group and type Ⅰ group (P＞0.05).The rates of esophagitis of type Ⅲ group and type Ⅱ group were higher than that of type Ⅰ group (71.4％,59.5％ and 14.3％,x2 =17.14 and 11.15,both P＜0.05),there was no significant difference between type Ⅲ group and type Ⅱ group (P＞0.05).There was one case of Barrett's esophagus in each group.The sensitivity and specificity of endoscopy in HH diagnosis were 91.4％ and 81.0％,respectively.Conclusions The distance between EGJ and hiatal pressure impression,continuously-open cardia,the His angle blunting,hiatal pressure impression of the gastric pouch and hernia sac were important indicators of HH endoscopic features.The sensitivity of endosocopy is higher than the spcificity of endosocopy in HH diagnosis.

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 To investigate the rehabilitative effect of low frequency repetitive transcranial magnetic stimulation (rTMS) on convalescing patients with Broca's aphasia. MethodsTwenty-eight patients with Broca's aphasia recovering from cerebral infarction were randomly divided into a stimulation group and a control group with 14 subjects in each.Patients in the control group accepted conventional drugs,speech rehabilitation and sham stimulation,while patients in the stimulation group were in given low frequency rTMS in place of the sham stimulation.Their speech performance was evaluated using the China Rehabilitation Research Center's aphasia examination (CRRCAE) pre-stimulation,post-stimulation and 90 days later. ResultsCompared with before treatment and with the controls,the speaking scores of the stimulation group increased significantly after treatment and also 90 days later. ConclusionLow frequency rTMS can not only improve the speech performance of Broca's aphasia sufferers in the short term,but it also plays a lasting role.It may thus have clinical application for patients with Broca's aphasia.