New tumor antigens are continuously being discovered due to the improvement of immunologic techniques.Whether dendritic cells induce immune activation or immune inhibition after they capture tumor antigens depends on the danger signals(GM-CSF,MCP1,and HSP) or the inhibitory signals(TGF-?,IDO,and iNOS) released by tumor cells.Under the regulation of danger signals,dendritic cells activate Th1 immune response and eliminate tumors;under the regulation of inhibitory signals,they activate Th2 immune response and can not effectively eliminate tumors.Progress in tumor immunotherapy mainly is manifested by antibody-based therapy,T cell-based therapy,and tumor vaccine-based therapy.To day at least 7 antibodies have been confirmed effective when combined with chemotherapeutic agents in treatment of tumors.Despite of the progress made in antibody therapy,discovery of new targets,development of new antibodies,and expanding of the application scope to more tumors still need intensive research efforts.The clinical effects of T cell-based therapy have not been satisfactory;most tumor vaccine-based therapies are in phase Ⅰ and Ⅱ clinical trial,and the outcomes of few phase Ⅲ clinical trials are not satisfactory,leaving more work to be done for improvement.As we understand more about the roles of antibody in immune surveillance,it will help to make immunotherapy of tumors a promising strategy.

Objective:To study MUC1 based cancer vaccine.Methods:MUC1 gene was inserted into pMAL-p2 vector and constructed recombinant pMAL-MUC1. MUC1-MBP fusion protein expression was induced by IPTG in E coli DH5? transformed by the recombinant pMAL-MUC1 .The fusion protein was analyzed by Western blot and purified by amylose affinity chromatography. The antiserum,T cell proliferation and CTL activity of spleen from C57 mice immunized by MUC1-MBP were determined respectively by ELISA,adding 3H-TdR and MTT.Results:Had successfully constructed pMAL-MUC1 expression vector,and purified MUC1-MBP and MBP. C57 mice immunized by MUC-MBP generated MUC1 specific antibody and CTL.The titer of polyclonal antibody to MUC1 was about 1∶5 760?3 221. CTL cytotoxicity to the MCF7 and lewis lung cancer cells respectively were at 47.7%?4.3% and 67.5%?6.5%.Conclusion:Human recombinant MUC1-MBP fusion protein activated T and B cell response in mice.The results suggested that the recombinant Muc1 may be used to develop protein vaccine against carcinoma.

Objective: To study the anti-tumor effect of recombinant human MUC1-MBP. Methods: The C57BL/6 mice were in oculated with MUC1-MBP by subcutaneous. MUC1 specific CTL activity of spleen were determined by MTT; The effects on prevention and treatment of tumor were observed by establishing lewis lung cancer-carrying mice. Results: The cytotoxicity of CTL from immunized mice to the MCF7 and Lewis lung cancer cells respectively was (47.7?4.3) % and (67.5 ?6.5) %; 5?10 5 lewis lung cancer cells following immunization were injected iv into C57BL/6 mice, after three weeks, the number of lung and tail tumor colonies was 51 and 5 for PBS and MUC1-MBP groups respectively and the suvival time was significantly delayed in immunized mice. The average volume of tumors in mice with MUC1-MBP was 386 mm 3 wherea control group was 4 000 mm 3 at tumor treating experiment. Conclusions: Recombinant human MUC1-MBP have significantly effects on prevention, treatment and inhibiting metastases of tumor. Our results suggested that the recombinant MUC1-MBP might be used to develop protein vaccine against human carcinoma.

Objective To evaluate the effectiveness of ultrafiltration technology in endotoxin removal from purified recombinant MUC1-MBP fusion protein (MUC1-MBP)and to demonstrate the effect of ultrafiltration on endotoxin removal.Methods CM Sepharose FF weak cation exchange (CM)(CM group), CM combined with Phenyl Sepharose 6 FF exchange (C6)(CM+C6 group),CM combined with ultrafiltration (CM+ultrafiltration group), and CM combined with C6 and ultrafiltration (CM+C6+ultrafiltration group)were used to purify the MUC1-MBP from E.coli. and remove endotoxin;the expression level of endotoxin was detected by Chromogenic End-point Tachypleus Amebocyte Lysate.Results There was a single band at the expected molecular weight of 62 000 by SDS-PAGE analysis.and the purity>96% by Quantity One analysis.The endotoxin levels in CM group and CM +C6 group were quite high and there was no significant difference between two groups (P>0.05 );the endotoxin level in CM+ultrafiltration group was significantly lower than that in CM group, and there was significant difference (P<0.01);the endotoxin level in CM+C6+ultrafiltration group was significantly decreased compared with CM+C6 group (P<0.01);there was no significant differences of endotoxin level between CM+ultrafiltration group and CM +C6+ultrafiltration group (P>0.05 ). Conclusion The effects of CM or CM combined with C6 on endotoxin removal are quite poor, especially C6;CM combined with ultrafiltration are quite effective on endotoxin removal,and ultrafiltration plays an important role in endotoxin removal.

Objective:To study the inductive effect of recombinant MUC1-MBP fusion protein combined with R848 on the immune activity of T cells in the mice,and to provide experimental basis for searching the suitable adjuvants of recombinant MUC1-MBP fusion protein.Methods:A total of 21 C57BL/6 mice were randomly divided into control group(treat with normal saline),MUC1-MBP+R848 group (treated with MUC1-MBP+R848),and MUC1-MBP+baillus calmette guerin(BCG) group(treated with MUC1-MBP+BCG)(n=7).After immunized for 4-7 d,the spleen tissue was taken and the spleen indexes of the mice in various groups were measured;the stimmulus index(SI) of the mice was detected by lymphocyte proliferation response;the levels of tumor necrosis factor-γ(TNF-γ) and interleukin-4(IL-4) were detected by ELISA method;the proprotions of T lymphocyte subsets in spleen cells were analyzed by flow cytometry.Results:Compared with control group,the spleen indexes,SI,and TNF-γ levels of the mice in MUC1-MBP+ R848 and MUC1-MBP+BCG groups were significantly increased (P<0.05 or P<0.01);the indexes metioned above of the mice in MUC1-MBP+ R848 were higher than those in MUC1-MBP+BCG group;the IL-4 levels had no significant difference(P>0.05).Compared with control group,the proprotions of CD3+T,CD4+T,and CD8+ T cells of the mice in MUC1-MBP+ R848 group and MUC1-MBP+BCG group were significantly increased(P<0.05 or P<0.01).Conclusion:Both MUC1-MBP fusion protein combined with R848 and BCG can induce the Th1 type of immune activity in the mice,and R848 is the potential candidate adjuvant for MUC1-MBP.

Objective: To study the protective function of Gardenia yellow (GY) against CCl 4 induced hepatotoxicity in mice.Methods: Healthy Kunming male mice, weighting (20?2) g, 10 per group were randomized into 5 groups:control group, CCl 4 injured group, low dose group(CCl 4 injured+0.1 ml GY solution), medium dose group(CCl 4 injured+ 0.2 ml GY solution) and high dose group(CCl 4 injured+0.4ml GY solution). GY solution was given i.g. 5 d prior to CCl 4 injury. Serum glutamate pyruvate transaminase(SGPT) and glutamic oxaloacetic transaminase(SGOT) and lactic dehydrogenase(LDH) activities were determined 18 h after CCl 4 injury. Hepatic malondialdehyde(MDA), glutathione(GSH) and liver index were also detected. Results: In GY treated groups, the increases of serum SGOT, SGPT, LDH activities and liver GSH were inhibited obviously. The elevations of MDA and liver index were prevented significantly. The lesions in liver lobule were ameliorated obviously. Conclusion: Gardenia yellow can protect against CCl 4 induced hepatotoxicity.

Objective: To investigate the snoring status of school-aged children in Beijing and explore the association of snoring and academic performance. Methods: A total of 7 925 children aged from 6 to 14 were selected from 15 primary and middle schools at 7 districts (Xicheng, Chaoyang, Changping, Shunyi, Fangshan, Huairou and Mentougou) in Beijing in 2015, using multi-stage stratified cluster random sampling method. The recruited children were asked to complete the Pediatric Sleep Questionnaire (PSQ) and a questionnaire related to sleep behavior. The multiplelogistic regression was used to analyze the association of snoring and academic performance. Results: A total of 794 (12.44%) children showed a decline in academic performance among 6 383 eligible respondentsfor data analysis. 580 (9.08%) children with snoring was identified, of which 333 and 247 were in frequency of 1-2 times per week and frequency of ≥3 times per week, respectively; 357, 170 and 53 were in snoring grade Ⅰ, grade Ⅱ and grade Ⅲ, respectively. Compared with the children without snoring, the OR (95%CI) for children with 1-2 times per week and ≥3 times per week was 1.363 (1.000-1.857) and 1.605 (1.135-2.269), respectively; and the OR (95%CI) for children with grade Ⅰ, grade Ⅱ and grade Ⅲ of snoring was 1.226 (0.893-1.683), 1.595 (1.062-2.397) and 2.31 (1.17-4.565), respectively. Conclusion There is a statistical relationship between snoring and the decline of academic performance. The decline of academic performance positively associated with increased frequency and grade of snoring.

Objective: To investigate the sleep quality of children in Beijing and to analyze the related factors. Methods: The data were collected from the survey of 3-14 years old children in 7 urban districts of Beijing in 2015. By using multi-stage stratified cluster random sampling method, 26 kindergartens and primary and secondary schools in 7 districts and counties, including Xicheng, Chaoyang, Changping, Shunyi, Fangshan, Huairou and Mentougou, were randomly selected, with a total of 11 420 children. Children′s sleep status was investigated with Children′s Sleep Questionnaire (PSQ), and the proportion of children with sleep quality problems when the PSQ score was greater than 7. Various sleep related factors were investigated with self-made questionnaire. A multilevel model was used to analyze the relationship between PSQ score and related factors. Results: The average PSQ score of the children was 3.60±2.69. The proportion of children with sleep quality problems was 8.87%(816/9 198). Multilevel model analysis showed that the younger the children, the higher the PSQ score (<6 years old vs. 6~12 years old vs. >12 years old: 3.94±2.58 vs. 3.58±2.66 vs. 3.30±2.84, F=33.015, P<0.001); male PSQ score higher than female (3.89±2.75 vs. 3.30±2.60, t=10.560, P<0.001); and snoring, obesity, father/mother snoring, playing games before bed, surfing the internet, eating and other factors were statistically related to PSQ. Conclusions Sleep quality of children in Beijing should not be neglected, especially preschool children with high PSQ scores. Parents should pay attention to children′s snoring problems and try to reduce some pre-sleep behaviors that may affect sleep quality.

Objective: To investigate the snoring status and related family factors of children from 3 to 14 years old in Beijing. Methods: From May to July, 2015, data of children from 3 to 14 years old were obtained from a status survey from 7 districts(Xicheng, Chaoyang, Changping, Shunyi, Fangshan, Huairou and Mentougou) in Beijing. A total of 11 420 children from 25 primary and middle schools were randomly selected. Pediatric Sleep Questionnaire (PSQ) and a self-administered questionnaire were carried out for the adopted children. Self-administered questionnaire included the snoring related family factors. Logistic regression was used to estimate the odd ratio(OR) with 95% confidence intervals for variables. Results: A total of 9 198 children meet the inclusion criteria and are analyzed in the study, of whom 901 (9.80%) were found with snoring behavior. The incidence of boys is higher than girls. Obese children take higher risk of snoring. Compared with younger children (≤6 years old), older children (≥12 years old) have a significantly lower risk of snoring (OR=0.464, 95%CI 0.368-0.585). There is no statistical association between full-term infants, infant feeding pattern, parental cigarette smoking and child snoring.The children with family history of snoring have a significantly higher risk of snoring occurrence. The educational background of mother is statistically related to children snoring (OR=1.241, 95%CI 1.058-1.457). Conclusions The incidence of children snoring in Beijing is 9.80%, male gender, obesity, and young age are all risk factors for children snoring. There is a significantly statistical relationship between snoring and related family factors, such as family snoring history and education experience.