Objective To modify the methods of operation and establishment of cerebral ischemia rat models made by thread occlusion. Method We randomly divided 120 male SD rats into a common group (n = 50), an improvement group (n = 60) and a sham-operated group (n - 10). Rats in the common and improvement groups were made into models using the common and improvement methods separately. All models were evaluated on the basis of physical sign indices and 2,3,5-triphenyltetrazolium chloride (TTC) staining. The TTC staining results were taken as gold standards. Then, we compared the achievement ratios of the two groups, and computed the sensitivity and specificity of every physical sign index based on these standards. The χ~2 or correction χ~2 test was used to compare the ratios. Results (1) The achievement ratio in the improvement group was significantly higher than that in the common group (71.67% vs. 52.00%, P = 0.034). (2) The sensitivity of evaluation for both common and improvement methods was 98.55%. However, the specificity of evaluation for the improvement method was significantly higher than that for the common method (100.00% vs. 40.00%, P =0.000). Conclusions The establishment achievement ratio and evaluation correctness of models are obviously elevated by modification of the operation and establishment methods.

Using objective structuned clinical exammation ,OSCE to access a new resident standardized training , analyze the result and perform stratifying management and teaching based on the result , which may improve the effi-ciency and effectiveness of resident standardized training .

ObjectiveTo observe the activity and safety of a novel combination therapy for patients with recurrent or refractory aggressive non-Hodgkin lymphoma (NHL).MethodsSix consecutive patients with recurrent or refractory aggressive NHL were treated with B-VIP regimen,boanmycin (5 mg/m2 on Days 1,4,8,12 and 15),vincristine (1.4 mg/m2 on Days 1,8 and 15),ifosfamide (1.2 g/m2 on Days 1,2,3 and 15,16,17) and prednisone (50 mg on Days 1 to 10),every 21 days.All the patients had received ≥5 cycles (average 8.3 cycles) of previous chemotherapy.ResultsSix patients (100 ％) were evaluable for response.The overall objective response rate was 66.7 ％ (4 patients),including 1 case complete (CR) and 3 cases partial responses.Myelosuppression was the most frequent serious complication of this regimen.ConclusionIn the current study,B-VIP was a highly active and safe combination therapy for patients with refractory disease with a poor prognosis or for patients with multiply recurrent aggressive NHL.

Objective To investigate the changes of invasion and migration potential of residual hepatocarcinoma cells after fractionated X-ray irradiation and its underlying mechanism.Methods HepG2 cells were exposed to X-rays (2 Gy × 10) and recovered for 30 days after irradiation to generate residual cells.The changes of cellular invasion and migration potential were detected in the residual and its control cells using a Transwell assay.The expressions of epithelial-mesenchymal transition(EMT)-related proteins of N-cadherin and Snail were detected by Western blot.HepG2 subcutaneous tumor models were established using nude mice that were divided into control and radiation group.In radiation group,the tumors were locally irradiated with a dose of 2 Gy per fraction daily,5 days per week for 2 weeks until the cumulative dose reached to 20 Gy.The growth of the tumor was observed,and on the day 39 after cell implantation (i.e.day 14 after radiation),the liver metastasis and the expression of N-cadherin in tumor with or without radiation were detected.Results The invasions and migrations of the residual cells and xenograft tumor were significantly enhanced in comparison with the control group (t =5.126,7.714,P ＜0.05).The expressions of N-cadherin and Snail in the radiation group were significantly higher than that in the control group (t =7.509,7.184,P＜0.05).In the HepG2 subcutaneous tumor model,the weight and volume of tumor in nude mice of the radiation group was significantly smaller than that in the control group (t =2.396,3.170,P ＜0.05),and the number of liver metastases in nude mice and the expression of N-cadherin in tumor were significantly higher than those in the control group (t =2.994,5.695,P ＜0.05).Conclusions Fractionated irradiation enhances the abilities of invasion and migration by inducing EMT in hepatocarcinoma cells,which provides new insights of the recurrence and metastasis of hepatocarcinoma after radiotherapy.

Objective To establish a method of detecting hepatitis B virus x antigen (HBxAg) and antibody to HBxAg (anti-HBx) and to demonstrate its clinical significance of HBxAg and anti-HBx in sera from patients of chronic hepatitis B (CHB),liver cirrhosis (LC) and hepatocellular carcinoma (HCC). Methods Full length HBx gene was cloned into pET30a(+),a prokaryotic expression vector,named pET30a-X.It was transformed into Escherichia coli BL21 (DE3),followed the fusion protein of HBx-His was induced by IPTG.The purified fusion protein was used to immunize rabbit as an antigen to generate polyclonal antibody to HBx protein.The method of enzyme-linked immunosorbent assay (ELISA) was established by using purified fusion protein and generated antibody,which was used to detect HBxAg and anti-HBx in sera from patients of CHB,LC,HCC and normal healthy people.Results The positive rates of HBxAg/anti-HBx were 8.7%/10.4% for CHB,17.9%/40.6% for LC,and 9.8%/34.4% for HCC, respectively.In statistics,the positive rates of anti-HBx in LC and HCC were higher than that in CHB (P

Orthotopic liver transplantation has proven to be effective in the treatment of a variety of life-threatening liver diseases, however, the limitations of donated organs available and long-term immunosuppression provided an impetus for developing alternative therapies. Cell replacement strategies have been one major effective approach for overcoming the obstacles of organ transplantation in recent years. The exogenous cells should be able to proliferate and differentiate into mature hepatic cells after grafting. Use of mature hepatocytes is also hampered by limited tissue source and inability to proliferate and maintain the function for a long term in vitro. Embryonic stem cells are immortal and pluripotent and may provide a novel cell source for potential cell therapy. This review summarizes the mechanisms of controlling early liver development and hepatic differentiation of visceral endoderm in embryoid bodies, and provides an overview of diverse differentiation systems in vitro and in vivo that were applied to hepatic research in recent years. Several studies have demonstrated that ES cell-derived hepatocytes can incorporate into liver tissue and function in vivo , but a few of them have shown complete restoration of liver function after transplantation into mice with liver diseases. Further studies should be made to exploit efficient methods and clinical applications of hepatocytes derived from ES cells in the future. In addition to clinical transplantation for treatment of liver diseases, ES cells can provide a valuable tool for drug discovery applications and study on of molecular basis of hepatic differentiation.
Animals ; Cell Differentiation ; physiology ; Cells, Cultured ; Embryonic Stem Cells ; cytology ; transplantation ; Hepatocytes ; cytology ; Humans ; Liver Diseases ; therapy

OBJECTIVETo explore the effects of Triptolide (TP) on TNFalpha-induced cell proliferation and expressions of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS) and their inducing products PGE2, NO in human rheumatoid arthritis synovial fibroblasts (RASF).

METHODSFibroblasts (RASF) were obtained from synovial tissue of patients with RA and were cultured in vitro. RASF were stimulated with TNFalpha(20 microg/L) in the presence or absence of TP(0 - 100 microg/L) for 20 h. The RASF proliferation was determined by (3)H-TdR incorporation, and the productions of PGE2 and NO in culture supernatants of RASF were detected with competitive ELISA and enzyme reduction of nitrate. Expressions of COX-2 and iNOS mRNA in RASF were analyzed by semi-quantitative RT-PCR. Expressions of COX-2 and iNOS protein were estimated by Western-blot method and cellular enzyme immunoassay in synovial fibroblasts. NF-kappaB activity in whole-cell extract of RASF was also measured by an ELISA-based method.

RESULTSTP (>20 microg/L) down-regulated markedly TNFalpha-induced COX-2 and iNOS mRNA and protein expression, and their inducing products PGE2 and NO of synovial fibroblasts. This effect was positively correlated with TP concentrations. NF-kappaB activity in TNFalpha-stimulated synovial cells was suppressed profoundly by TP treatment (IC(50) approximately 35microg/L). The activity of NF-kappaB was correlated with the levels of COX-2 and iNOS expression in TNFalpha-stimulated RASF. No change was observed in proliferation of synovial cells after treatment of TP.

CONCLUSIONTP could significantly down-regulate TNFalpha-induced COX-2, iNOS expression and production of PGE2, NO in human RASF, which is associated with the suppression of NF-kappaB activity.

Arthritis, Rheumatoid ; drug therapy ; metabolism ; Cyclooxygenase 2 ; Diterpenes ; pharmacology ; Epoxy Compounds ; Fibroblasts ; metabolism ; Gene Expression Regulation ; drug effects ; Humans ; Isoenzymes ; analysis ; genetics ; Membrane Proteins ; NF-kappa B ; metabolism ; Nitric Oxide Synthase ; analysis ; genetics ; Nitric Oxide Synthase Type II ; Phenanthrenes ; pharmacology ; Prostaglandin-Endoperoxide Synthases ; analysis ; genetics ; RNA, Messenger ; analysis ; Synovial Membrane ; cytology ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology

Objective:To compare the efficacy of posterior atlas uniaxial and polyaxial screw instrumentation and fusion with bone graft for Gehweiler type IIIb atlas fracture.Methods:A retrospective cohort study was performed to analyze the clinical data of 36 patients with Gehweiler type IIIb atlas fracture admitted to Henan Provincial People′s Hospital from January 2015 to October 2020. There were 29 males and 7 females, with age range of 23-82 years [(48.8±15.5)years]. All patients were treated with posterior atlas screw-rod internal fixation and fusion with bone graft, of which 14 received atlas uniaxial screw internal fixation (uniaxial screw group) and 22 received atlas polyaxial screw internal fixation (polyaxial screw group). The operation time and intraoperative blood loss were compared between the two groups. The atlas fracture union rate and atlantoaxial posterior arch bone fusion rate were compared between the two groups at 3 months and 6 months after operation. The anterior atlantodental interval (ADI), basion-dens interval (BDI) and lateral mass displacement (LMD) were compared between the two groups to evaluate the reduction of fracture fragments before operation, at 1, 3, 6 months after operation and at the last follow-up. At the same time, the visual analogue scale (VAS) and neck dysfunction index (NDI) were compared between the two groups to evaluate neck pain and functional recovery. The postoperative complications were observed.Results:All patients were followed up for 12-44 months [(27.2±9.9)months]. There was no significant difference in operation time or intraoperative blood loss between the two groups (all P>0.05). The atlas fracture union rate and atlantoaxial posterior arch bone fusion rate were 85.7% (12/14) and 78.6% (11/14) in uniaxial screw group at 3 months after operation, insignificantly different from those in polyaxial screw group [72.7% (16/22) and 77.3% (17/22)] (all P>0.05). All patients in the two groups achieved bone union and fusion at 6 months after operation. There was no significant difference in ADI between the two groups before and after operation (all P>0.05). The BDI in the two groups did not differ significantly before operation ( P>0.05), but a significantly higher value was found in uniaxial screw group at 1, 3, and 6 months after operation and at the last follow-up [(5.9±1.3)mm, (5.8±1.3)mm, (5.9±1.2)mm and (5.8±1.2)mm] than in polyaxial screw group [(3.1±0.6)mm, (3.1±0.6)mm, (3.1±0.6)mm and (3.1±0.6)mm] (all P<0.01). The two groups did not differ significantly before operation ( P>0.05), but LMD at 1, 3, and 6 months after operation and at the last follow-up was (1.6±0.8)mm, (1.5±0.8)mm, (1.5±0.7)mm and (1.5±0.9)mm in uniaxial screw group, significantly lower than that in polyaxial screw group [(4.8±1.6)mm, (4.6±1.6)mm, (4.9±1.6)mm and (4.9±1.6)mm] (all P<0.01). There was no significant difference in VAS between the two groups before operation ( P>0.05). The VAS at 1, 3, and 6 months after operation and at the last follow-up was 3.0(3.0, 4.0)points, 2.0(1.0, 2.0)points, 1.0(0.8, 2.0)points and 1.0(0.0, 1.3)points in uniaxial screw group and was 3.5(3.0, 4.0)points, 2.0(2.0, 3.0)points, 2.0(1.0, 2.0)points and 2.0(1.0, 3.0)points in polyaxial screw group. In comparison, the VAS scored much lower in uniaxial screw group than in polyaxial screw group at 6 months after operation and at the last follow-up (all P<0.01). There was no significant difference in NDI between the two groups before operation ( P>0.05). The NDI at 1, 3, and 6 months after operation and at the last follow-up was 34.9±6.3, 23.4±6.2, 13.9±2.7 and 9.4±2.8 in uniaxial screw group and was 33.2±6.1, 24.4±6.3, 18.1±4.1 and 12.7±3.2 in polyaxial screw group, showing a significantly lower NDI in uniaxial screw group than in polyaxial screw group at 6 months after operation and at the last follow-up (all P<0.01). The complication rate was 21.4% (3/14) in uniaxial screw group when compared to 22.7% (5/22) in polyaxial screw group ( P>0.05). Conclusion:For Gehweiler type IIIb atlas fracture, both techniques can attain atlas fracture union and atlantoaxial posterior arch bone fusion, but the posterior atlas uniaxial screw instrumentation and fusion is superior in reduction of atlas fracture displacement and lateral mass separation, neck pain relief and functional improvement.

Objective:To investigate the polymorphisms at HLA-A,-B,-DRB1 loci in Dalian Han population. Methods: A total of 10 000 unrelated marrow donors who live in Dalian were genotyped by SBT and SSO methods. Haplotype frequencies and linkage dis-equilibrium values were calculated by ARLEQUIN software,and DA genetic distances between populations were calculated by poptree2 software. Results: A total of 18 HLA-A alleles, 32 HLA-B alleles and 13 HLA-DRB1 alleles were found in Dalian Han population. HLA-A?02 (31. 65% ),B?40(14. 84% ) and DRB1?15(15. 82% ) occurred most frequently. A?30-B?13-DRB1?07 (4. 56% ) was determined to be the most common three-locus haplotype and the second predominant haplotype was A?02-B?46-DRB1?09 ( 2. 43% ) . A ?30-B ?13 ( 6. 00% ) and B ?13-DRB1 ?07 ( 59. 89% ) were the most common two-locus haplotypes. Moreover,A?33-B?58 and B?13-DRB1?07 were strongest haplotypes with the linkage disequilibria values 0. 336 6 and 0. 665 1,respectively. In China,the closest genetic distances were found with Heilongjiang (0. 001) followed by Jilin (0. 002) and Shandong (0. 002),the furthest was found with Taiwan (0. 047). Compared with other populations worldwide,the closest genetic distances were found with Thailand (0. 029) and Korea (0. 03),the furthest was found with Italy (0. 183). Conclusion: Dalian Han population had rich polymorphism at HLA-A,-B,-DRB1 loci,and the distribution of HLA-A,-B and-DRB1 was in line with the charac-teristics of the northern population.

Objective To detect the feasibility of magnetically labeled swine bone marrow mesenehymal stem cells(MSCs)with SHU 555A combined with poly-L-arginine(PLL),under MR imaging in vitro and in vivo.Methods Swine mesenehymal stem cells were isolated and culture-expanded 3 passages in vitro,then magnetically labeled by incubation with SHU 555A(25?g Fe/ml,Resovist,Schering)for 24 hours with 750 ng/mL poly-L-lysine(PLL;average MW_275 kDa)added 1 hour before incubation.Cellular iron incorporation and detention at 0 d,4 d,8 d,12 d,16 d,20 d after labeling was qualitatively assessed using Prussian blue and quantified at atomic absorption spectrometry.Cell viability was assessed by trypan-blue exclusion test.Cell suspensions underwent MR imaging with T_1-and T_2-weighted spin-echo and fast field-echo sequences on a clinical 1.5 T MR system.At last,1?10~6 SHU 555A labeled and unlabeled MSCs were transextracardially implanted into the infracted and normal myocardium approximately 2 week following the ligation of left anterior descending coronary artery in 1 swine respectively,and finally performed 1.5-T MRI within 1 week after infarction.Results①Intracytoplasmic particles stained with Prussian blue stain were detected for all cells with mean cellular iron content of(13.13?2.30)pg per cell.With division of stem cells, the stained particles decreased gradually with iron content(0.68?0.20)pg per cell.at 16 days after labeling, approximately to the prelabeled baseline values.(0.21?0.06)pg per cell(P＞0.05).The viability of the labeled cells at various time points were not significantly different with that of nonlabeled cells(P＞0.05).②MR images showed signal intensity changed most obviouly in T2*WI in vitro.The percentage change of signal intensity increased with increasing cell numbers,and decreased with the time.As few as 5?10~4-1?10~5 cells could be detected by using this approach.③Two injected sites containing MR-MSCs were detected in vivo,presentingas low signal intensity areas with the T_2*WI scanning sequence.Conclusion Swine bone marrow MSCs can be labeled with SHU555A-PLL and depicted with a standard 1.5-T MR imager in vitro and in vivo.(J lntervent Radiol,2007,16:115-121)