Objective To investigate the effect of high mobility group box-1 protein (HMGB1) on immune function of peritoneal macrophages in mice. Methods The peritoneal macrophages were isolated from female BALBc mice in vitro. The macrophages were stimulated with different doses of HMGB1 (1, 10, 100, 1 000ng/ml) for 6 hours, or with 100ng/ml HMGB1 for different duration (0, 6, 12, 24, 48, 72 hours). The phagocytosis ability of macrophages was determined by the neutral red dye uptake assay and optical densities of samples were read at 540nm. The cytotoxicity to L1210 cells was measured with MTT assay and was expressed by absorbance at 570nm. The chemotaxis was assayed by using the Costar Transwell cell culture chamber inserts and the cells on the lower surface of the transwell membrane were fixed and counted. The cells were harvested at the indicated time points, fixed, and stained for indirect immunofluorescence, then the expressions of I-Ak MHC class Ⅱ alloantigen on macrophages were determined with flow cytometry. Results The effects of HMGB1 on phagocytic activities, cytotoxicity, chemotaxis and I-Ak antigen expression of macrophages were enhanced in a dose-dependent manner, starting at doses as low as 1ng/ml and with a maximal response at 100ng/ml (P

Objective Intranuclear forkhead/winged helix transcription factor p3(Foxp3) plays a key role in T cell-mediated immunosuppression.The present study was performed to investigate the effects of high mobility group box-1 protein(HMGB1) on Foxp3 gene as well as protein expressions in splenic regulatory T cells(Tregs) and their potential regulating mechanisms in mice.Methods CD4+CD25+Tregs isolated from the spleens of male BABL/c mice by magnetic beads were seeded on 96-well(1?105 cells/well) cell culture plates coated with anti-CD3(1 ?g/ml) and soluble anti-CD28(1 ?g/ml),and the cells were stimulated with HMGB1 at various intervals or at different concentrations.After being stimulated,the Foxp3 mRNA/protein expressions in the Tregs were determined.The time-dependent and dose-dependent responses between HMGB1 and intranuclear Foxp3 expression were analyzed by flow cytometry,and the expressions of Foxp3 mRNA of Tregs were analyzed by quantitative PCR of SYBR GREEN.Results After stimulation with HMGB1,the intranuclear Foxp3 protein and mRNA expressions of splenic Tregs in mice were markedly down-regulated in 24 h to 72 h(P

Objective:To study the effect of interleukin-18 on transdifferentiation in renal tubular epithelial cells(TECs).Methods:Human proximal tubular epithelial cell line(HK-2) was cultured in vitro.TECs were exposed to different concentrations(0,0.1,1,10 and 100 ng/ml) of IL-18 for 24,48 and 72 hours.At the end of each incubation,the expressions of the ?-SMA and TGF-?_1 mRNA were assessed by RT-PCR,the rate of ?-SMA expressing TECs was assessed by immunocytochemistry,and the expression of the ?-SMA protein was assessed by Western blotting.Results:(1)The expressions of ?-SMA and TGF-?_1 mRNA were increased significantly by a dose- and time-dependent manner when TECs were exposed to IL-18(P

Objective To discuss the methods and significance of exposing recurrent laryngeal nerve in thyroidectomy.Methods 328 recurrent laryngeal nerve were dissected and exposed.In 216 procedures(3 nerve were not recurrent,and all were in right lateral.),150(45.73%)nerve were dissected by infra thymid gland route,178(54.27%)by lateral thyroid gland route.Results There was no permanent injury of recurrent laryngeal nerve.It took 5 minutes on average to reveal nerve by infra thyroid gland route and there were 5 cases (2.00%)of temporary recurrent laryngeal nerve injury.It took 3 minutes on average to reveal nerve by lateral thyroid gland route,and there were 2 cases(1.12%)of temporary recurrent laryngeal nerve injury.Conclusions It is safe and convenient to expose recurrent laryngeal nerve during operation.Lateral route has advantages of less time consuming,minimal injury and less bleeding.

Objective To investigate the clinical efficacy of transarterial chemoembolization (TACE) combined with sorafenib for the treatment of inoperable hepatocellular carcinoma (HCC), and to discuss the influence of treatment interval on the survival time. Methods During the period from July 2008 to May 2011 at authors’ hospital, a total of 50 patients with inoperable HCC were treated with TACE together with sorafenib. The treatment intervals between each TACE procedure were recorded. The results were analyzed. Results Up to Dec. 31, 2011, the median follow-up time of the 50 patients was 310 days. The mean interval between TACE treatments was 69 days before the combination treatment was employed , while the mean interval was 112 days after the combination treatment started , and the longest interval was 648 days. Conclusion Combination treatment TACE with sorafenib can remarkably prolong the treatment interval in patients with inoperable HCC, thus the patient can get more survival benefits.

OBJECTlVE To study the effect of ganoderma poIysaccharide( GLP)on zebrafish ( Danio rerio)survivaI,deveIopment and senescence. METHODS A p53-knockout zebrafish modeI was estabIished by injecting of morphoIino phosphorodiamidate(mO)into normaI zebrafish eggs in the singIe ceII deveIopment stage. Different concentrations of GLP 1,2 and 3 g·L-1 were used to treat both wiId type and p53-knockout zebrafish embryos 8 h post fertiIization(8 hpf)under 28℃ for 3 d. The survivaI and maIformation rates were caIcuIated after 72 hpf,and the effect of GLP on ceII senescence was evaI-uated by senescence associated β-gaIactosidase(SA-β-gaI)staining both in wiId type and p53-knockout zebrafish embryos. In addition,the differentiaI gene expression of cancer inhibitor gene(p53),teIomer-ase reverse transcriptase( TERT),murine doubIe minute2( mdm2)and p21 was examined by RT-PCR both in wiId type and p53-knockout zebrafish embryos after treatment with different concentrations of GLP. RESULTS GLP 1 and 2 g·L-1 had no effect on deveIopment of wiId type zebrafish embryos,but after GLP 3 g·L-1 treatment,most of zebrafish embryos dispIayed maIformation and the survivaI rate was onIy 48.6%. GLP 1,2 and 3 g·L-1 had no effect on survivaI and deveIopment of p53-knockout zebrafish embryos. After GLP 2 g·L-1 treatment for 3 d,the SA-β-gaI staining positive rate of wiId type zebrafish embryos was reduced compared with controI group(P﹤0.01),whiIe there was no significant change in p53-knockout zebrafish embryos. The resuIts of RT-PCR showed that GLP 2 g·L-1 treatment depressed p21 and p53 gene expression(P﹤0.05,P﹤0.01),and had no effect on mdm2 and TERTgene expres-sion of wiId type zebrafish embryos. For p53-knockout zebrafish embryos,the TERT,mdm2,p21 and p53 gene expression dispIayed no significant difference after GLP 2 g·L-1 treatment. CONCLUSlON GLP 2 g·L-1 may improve senescence of wiId type zebrafish embryo ceIIs. GLP ≥3 g·L-1 may Iead to death and abnormaI deveIopment of wiId type zebrafish embryos. The improvement of GLP on senescence of wiId type zebrafish embryo ceIIs might be mediated by its down-reguIation of p21 and p53 gene expression.

Objective To investigate the application of C- arm CT imaging technique in evaluating the efficacy of transarterial chemoembolization(TACE) for hepatocellular carcinoma(HCC). Methods During the period from Sep. 2012 to Mar. 2013 at authors’ hospital, C- arm CT scanning was employed during the performance of TACE in 80 patients with HCC. The residual active tumor lesions, Lipiodol deposition and the embolization of the feeding arteries after TACE therapy were assessed by C- arm CT plain scan images and the images of early artery phase and parenchymal phase, as well as the post- processing images. The efficacy of TACE was thus evaluated. Results A total of 139 hepatic tumors were detected in 80 patients by C- arm CT imaging. C- arm CT scanning performed immediately after TACE showed that satisfactory result was obtained in 128 lesions of 75 patients(128/139, 92.9%). Fewer, medium and more residual active tumors were seen in 78, 29 and 21 lesions respectively, while dense, moderate and thin Lipiodol deposition was seen in 64, 39 and 25 lesions respectively. After TACE therapy obvious decrease in blood supply was demonstrated in 101 lesions, while no obvious decrease in blood supply was seen in 27 lesions. Conclusion C- arm CT scanning after TACE for patients with HCC can conveniently, accurately and comprehensively reveal the residual active tumors, Lipiodol deposition and embolization status of feeding arteries. Therefore, this technique should be regarded as an important means to evaluate the efficacy of TACE.

Objective To evaluate the value of the mRECIST criteria in assessing the efficacy of transcatheter arterial chemoembolization(TACE) combined with sorafenib in the treatment of hepatocellular carcinoma (HCC).Methods A total of 35 patients who were treated with a combination of TACE and sorafenib for unresectable hepatocellular carcinoma fulfilled the mRECIST and RECIST criteria in our hospital from June 2011 to November 2012.Enhanced CT and/or enhanced MRI were used before (baseline) and after (3 month reexamination) combination treatment in our hospital.The mRECIST and RECIST criteria were used to evaluate the efficacy,and these efficacy assessments were compared.Results In the RECIST criteria,complete remission (CR) was 0％,partial remission (PR) was 2.9％,stable disease (SD) was 85.7％％,and progressive disease (PD) was 11.4％％.In the mRECIST criteria,CR was 8.6％,PR was 51.4％,SD was 34.3％,and PD was 5.7％.For the RECIST criteria,the objective response rate (CR+-PR) was 2.9％,the disease control rate (CR+PR+SD) was 88.6％,and the disease progression rate was 11.4％.For the mRECIST criteria,the objective criteria was 60％,the disease control rate was 94.3％,and the disease progression rate was 5.7 ％.The difference between the efficacy assessment results of mRECIST and RECIST was statisti cally significant(P＜0.001).Conclusion The mRECIST criteria can evaluate the efficacy of target le sions based on viable tumors,which is more adaptive to TACE and targeted drugs with new mecha nisms.

Objective To confirm the transmission of Toxoplasma gondii by semen and to investigate the impact of vaginal status on the transmission of T. gondii in female rabbits. Methods Sixteen male rabbits were infected with T. gondii by intraperitoneal injection each with 1 ×105 RH tachyzoites. Eight rabbits died in 8-14 d after infection.Artificial vagina was used to collect semen from male rabbits weekly before and after infection for 8 weeks. If more than 2 portions of semen from 8 survived male rabbits were collected after infection, the collected semen was mixed weekly for later use. Twenty-seven female rabbits were divided into 4 groups: group 1 with normal vagina (7 rabbits), group 2with wounded vagina (7), group 3 with trichomonas vaginitis (7) and group 4 with colpomycosis infection (6). Tachyzoites were found in mixed semen digested by trypsinase, and were used for endovaginal artificial insemination to female rabbits by uterine cavity tube once a week for 8 consecutive weeks. 2-3 d after every insemination, 2 ml blood was collected from helix vein of each rabbit, and stored at -40 ℃ for use. Anti-T. gondii antibody was examined by ELISA and the B1 gene of T. gondii was detected by PCR. Results Anti-T. gondii antibody was detected in some rabbits (2, 3, 1, and 1 rabbits from each of the groups respectively) on the 16th day after the first insemination. The positive rate of ELISA was 25.9%. The amplification of B1 gene (200 bp) by PCR appeared positive from the blood samples on the 3rd day after the first insemination and the last positive one was proved on the 51th day after the first insemination.Number of positive samples was 2, 1, 3 and 1 in the 4 groups respectively, with an overall PCR positive rate of 18.5%.Only 3 of the 27 rabbits were positive by both ELISA and PCR. Conclusions T. gondii can be transmitted by semen and the health status of vagina shows no impact on it.

Objective Thoracic cavity fistula following esophagus carcinoma resection is a serious complication with a high mortality.This study aims at a better therapy for thoracic cavity fistula following esophagus carcinoma resection by summarizing the ex-perience with the four-tube strategy ( jejunal fistula tube, stomach tube, chest drainage tube, and nasal fistula tube) in the treatment of the complication. Methods We retrospectively analyzed the clinical data about 62 cases of thoracic cavity fistula following esopha-gus carcinoma resection, 35 treated with the four-tube strategy ( treatment group) and the other 27 with the three-tube ( stomach tube, chest drainage tube, and nasal fistula tube) method ( control group) .We compared the hospital days, wound healing time, mortality, and incidence of anastomotic stenosis at 6 months after operation between the two groups of patients. Results Compared with the controls, the treatment group showed remarkable decreases in the hospital days (P<0.05), wound healing time (P<0.05), and mortality (P<0.05), but no statistically significant difference was observed in the incidence rate of anastomotic stenosis at 6 months after operation between the two groups of patients ( P >0.05 ) . Conclusion Compared with the three-tube method, the four-tube strategy has the advantages of shorter healing time and lower mortali-ty, and therefore is preferable for the treatment of thoracic cavity fis-tula following esophagus carcinoma resection.