Expression of CD4+CD25+CD127Low regulatory T cells and cytokines in peripheral blood of patients with primary liver carcinoma.

Objective: To assess the clinical utility of the ratio of CD4+CD25+CD127low regulatory T cells (Tregs) in subjects at high risk of HCC, investigate the relationship between the percentage of Tregs and the expression of transforming growth factor (TGF)-ß1 and interleukin (IL)-10 in patients with hepatocellular carcinoma before and after treatment. Methods: Peripheral venous blood was collected from patients with liver cancer before and after treatment. The proportion of CD4+CD25+CD127low Tregs was detected by flow cytometry. The levels of TGF-ß1 and IL-10 in serum were detected by enzyme-linked immunosorbent assay, and were compared with healthy subjects as a control group. Results: The proportion of CD4+CD25+CD127low to CD4+T lymphocytes in patients with hepatocellular carcinoma was significantly higher than that in healthy controls (P<0.01). The proportion of CD4+CD25+CD127lowTregs, whose AUC of ROC curve was 0.917, could effectively separate the HCC patients from the healthy subjects with a diagnostic sensitivity of 90%, specificity of 80%. The proportion of CD4+CD25+CD127low to CD4+T lymphocytes and the levels of TGF-ß1 and IL-10 in patients with hepatocellular carcinoma after the operation and chemotherapy were significantly lower than those before treatment (P<0.05).The proportion of CD4+CD25+CD127lowTregs was positively correlated with the concentrations of TGF-ß1 and IL-10 before and after treatment of primary liver cancer (P<0.05). Conclusion: CD4+CD25+CD127lowTregs may be a significant predictor of HCC biopsy outcome and play an inhibitory role on effector T cells by regulating cytokines.

Efficient production of human zona pellucida-3 in a prokaryotic expression system.

The zona pellucida-3 (ZP3) protein plays a pivotal role in oocyte and gamete development. We aimed to produce a recombinant ZP3 peptide using the Escherichia coli secretory system and apply it to a protein chip for detecting anti-ZP3 antibodies. The ZP3 gene was cloned into the pHOA downstream of the phoA promoter and transformed into E. coli YK537. Recombinant ZP3 was secretory expressed by decreasing the inorganic phosphate concentration. Then, rZP3 was purified and coated onto a protein chip, which was used to detect AZP3A in serum samples from 63 infertile patients. The area under the receiver operating characteristic curve was 0.934. The results, in terms of AZP3A detection, of the rZP3-coated protein chip were consistent with those of the ELISA kit. Therefore, our protein chip assay has potential for diagnosis of infertility due to AZP3A, and represents a less costly and simpler assay for clinical and research applications.

ICOS signal facilitates Foxp3 transcription to favor suppressive function of regulatory T cells.

Inducible costimulator (ICOS) plays an important role in the suppressive immunity mediated by regulatory T cells (Tregs), but the molecular regulation mechanism is not well known. Here we performed a study to explore the possible mechanism by which ICOS regulates the suppressive functions and survival of Tregs. This study showed that both the ICOS and CD28 signal could promote the survival of Tregs. However, ICOS but not CD28 improved the suppressive function of Tregs. Mechanistic studies demonstrated that ICOS could induce the transcription activity of Foxp3, by facilitating the nuclear factor of activated T cells (NFAT): Foxp3 over NFAT: activator protein 1 (AP-1). The results of Q-PCR showed that AP1 downstream regulatory genes (IL-2 and IL-6) were down-regulated, and Foxp3 downstream regulatory genes (IL-4, IL-10 and TGF-ß) were up-regulated. Further, ICOS promoted anti-apoptosis may be by activating protein kinase B (Akt) signal. These findings demonstrated that ICOS signal could facilitate Foxp3 transcription in favor of survival and suppressive function of Tregs.

Depletion of regulatory T cells by anti-ICOS antibody enhances anti-tumor immunity of tumor cell vaccine in prostate cancer.

ICOS+Treg cells exert important immunosuppressive effects in tumor immunity. We adopt a combination approach of ICOS+Treg cells depletion with tumor cell vaccine to evaluate anti-tumor immunity in mouse prostate cancer model. Streptavidin (SA)-mGM-CSF surface-modified RM-1 cells were prepared as the vaccine and the mouse subcutaneous prostate tumor model was used to evaluate the immunity. Tumor growth, flow cytometry, immunohistochemistry, immunofluorescence and enzyme linked immunosorbent assay (ELISA) were performed to evaluate the therapeutic effects. Our results demonstrated that SA-mGM-CSF vaccine was prepared successfully and tumor growth was inhibited. The tumor size in the combination group was much smaller than that in the vaccine with IgG mAb group. The portions of dendritic cells, CD8+ and CD4+T cells in the mice blood and tumor tissues were increased after treatment with vaccine. There were more immune-suppressing Tregs infiltrated into tumor after treatment with tumor cell vaccine, and ICOS blocking could deplete the infiltrated Tregs, and T lymphocytes increased more dramatically in the combination therapy group. The concentrations of interferon-γ were increased in all vaccine group, the concentrations of Interleukin-10 and Interleukin-4 were much lower in the combination group. Our study demonstrated that ICOS blocking could deplete the tumor-infiltrated ICOS+Treg cells. Combining GM-CSF surface-modified RM-1 cell vaccine with Anti-ICOS antibody could induce better antitumor immunity than a vaccine alone.

[Expression of Livin in laryngeal squamous cell carcinoma and relationship with bFGF].

OBJECTIVE: To detect the roles of Livin and its relationship with bFGF in laryngeal squamous cell carcinoma (LSCC) through observing the expression of Livin and bFGF in laryngeal squamous cell carcinoma. METHOD: Expression of Livin and bFGF in 41 cases of LSCC (11 cases with lymph node metastasis) and 20 cases of normal soft palate mucosa were detected by immunohistochemistry. RESULT: Livin were positively detected in 29 (70.73%) cases of LSCC and negatively detected in all normal soft palate tissue. The positive rate of Livin was higher in LSCC than that in normal soft palate tissue and the expressions were significantly associated with lymph node metastasis status (P < 0.05) but not with histological grade, clinical stage and age (P > 0.05). The expression of Livin was positively correlated with the expression of bFGF. CONCLUSION: The elevated expression of Livin in LSCC might play an important role in the development of laryngeal squamous cell carcinoma and bFGF might be involved in the process.

[Bacteriological study on adult chronic sinusitis operated after endoscopic sinus surgery].

OBJECTIVE: To explore the bacteria isolated from middle nasal meatus, maxillary sinus, ethmoid sinus and postoperative cavity of patients with chronic rhinosinusitis and their characteristics of antibiotic resistance. METHODS: Eighty-seven patients with chronic rhinosinusitis were operated on by ESS to obtain the pus specimen for bacterial culture and antibiotic susceptibility test, before and 1 month, 3 months and 6 months after operation. RESULTS: Totally 645 strains (26 species) of bacteria were detected in 464 specimens [total positive rate was 78.9% (366/464)], in which aerobic bacteria was 95.3% (615/645). Gram negative bacteria and gram positive bacteria were 51.2% (330/645) and 48.8% (315/645), respectively. There was supernumerary tendency in detectable rate of gram negative bacteria isolated from postoperative groups. The main pathogens of postoperative patients were gram negative bacteria, with Enterobacter aerogenes, Pseudomonas aeruginosa and Hemophilus influenza occupying the first 3 places. The detectable rate of multiple drug resistance bacteria in postoperative group was much higher than preoperative groups, in which gram negative bacteria was the most, especially for Pseudomonas aeruginosa. There was significant difference in beta-lactamase detectable rate of the bacteria isolated from the delayed recovery group and the preoperative group (chi2 = 4.85, P < 0.05), Enterobacteriaceae occupied the first place among the beta-lactamase detectable bacteria isolated from the delayed recovery group. There was no significant difference in detectable rate of kinds of bacteria isolated from recovery group and control group. CONCLUSIONS: The main pathogens of patients with chronic rhinosinusitis are multiple drug resistance gram negative bacteria after operation, in which Pseudomonas aeruginosa occupies the first place. Gram negative bacteria are becoming the main opportunity pathogenic bacteria, which shows antibiotic resistance. microbial population of postoperative cavity from recovery group are becoming balanced.

[Correlation between the prognosis and the expression of survivin in surgical margins of laryngeal squamous cell carcinoma].

OBJECTIVE: To investigate the relationship between the local recurrence and the five years survival rate and the expression of Survivin in surgical margins of laryngeal squamous cell carcinoma. METHOD: Immunohistochemical technique (S-P) and image analysis were used to detect the expression of Survivin in the pathological negative margin specimen and primary site of cancer of 54 patients. RESULT: (1) Expression of Survivin was detected in 66. 7% (36/54) in laryngeal carcinoma, 40.7% (22/54) in surgical margins and negative in normal mucous membrane of palatoglossal pillar. A significant difference was observed for positive index(PI) in three groups (P < 0.01). (2) PI of Survival in surgical margins of patients with the local recurrence was higher than those with no local recurrence (P < 0.01). (3) The 5-year survival rate of patients with Survivin positive margin was lower than those with negative margin (29.3% vs 58.5%, P < 0.01). CONCLUSION: The expression of Survivin in surgical margins may predict the possibility of local recurrence and poor prognosis in a certain extent for patients with laryngeal carcinoma, which may guide the treatment after surgery.