Objective To clarify whether the regulatory effect of all-trans-retinoic acid (atRA) on the antigen presentation function of dendritic cell(DC) is tightly associated with NF-κB signaling pathway.Also to clarify atRA mainly affected differentiated DC or influence the differentiation procedure from bone marrow cell to DC.Methods MOG35-55 immunized C57BL/6J mice received administration of atRA or not,splenic DC and CD4 cells isolated from immunized mice were subjected to in vitro cross-culture,treated with IL-12 and IL-23 respectively.Th1 and Th17 polarization of stimulated CD4 cells were determined by intracellular staining and FACS analysis,while the production of their corresponding cytokines,IFN-γ and IL-17,were measured by ELISA.Bone marrow cells were isolated from the femurs of the na(i)ve mice,treated with IL-4 and GM-CSF with or without synergistic RA treatment.MHC Ⅱ and CD11c molecules on the DC were assayed by immune staining and FACS analysis,their antigen presentation functions were decided by the proliferation and cytokine production of the Th1 effecter cells stimulated by antigen pulsed DC.To investigate the status of the NF-κB signaling pathway,the amount of phospho-Ser536 NF-κB p65 in the whole DC lysate and total NF-κB p65 in the nucleus were detected by Western blot.Finally,selective RA receptor antagonists were studied to figure out which receptor was majorly involved in the regulatory effect of RA on DC.Results Splenic DC from RA treated mice showed significantly decreased function of presenting antigen to stimulate CD4 cell polarization and cytokine production.Compared with untreated control,RA in vitro treated DC showed decreased expression of MHC Ⅱ and CD11c on its cell surface,which was accompanied with depressed function of stimulating Th1 cell proliferation and IFN-γ production.Further study revealed that RA mainly affect the differentiation procedure from BMC to DC,however,has no significant effect on differentiated DC as the aspects of MHC Ⅱ,CD11c expression,responder cell proliferation and cytokine production were evaluated.Decreased amount of phosphor-Ser536 NF-κB p65 in the whole cell lysate and total NF-κB p65 in the nucleus was investigated in RA treated DC,and decreased antigen presentation function of RA treated DC always came together with diminished activation of NF-κB signaling pathway.Finally it was demonstrated that RARβγ antagonist,but not RARα antagonist,could entirely block the RA effect on DC.Conclusion Retinoic acid inhibit the differentiation of DC as well as decrease its antigen presentation function,which might be resulted from the inactivation of NF-κB p65 signaling pathway and mediated by RARβγ.

Objective:To construct an E.coli expressing system of human interferon-?(IFN-?).Methods:Extracted DNA from human blood and PCR human IFN-?, cloned the human IFN-? gene into plasmid T-easy and pBV-220. Expressed human IFN-? in E.coli DH5?, the expressing product was analysed by SDS-PAGE, Western blot and anti-virus capacity test.Results:DNA sequence analysis showed the recombinant plasmid pBV- IFN-? contained human IFN-?. SDS-PAGE and Western blot proved that there were hIFN-? in E.coli DH5? after temperature inducing and the expressing product has anti-virus activity.Conclusion:A human IFN-? E.coli expressing system was constructed successfully, and the recombination human IFN-? has anti-virus activity.

Objective:To establish a DNA typing method for HLA-I antigens with medium resolution method by DNA chip technique.Methods:The chip was made with specific medium distinguish-typing probes designed according to gene frequency of HLA-I alleles from Northern Chinese. Unsymmetrical PCR was used to amplify HLA-I exon2,3,and then the PCR products labeled and hybridized with probes on the chip.Typing of HLA-I was certified by scanning the hybridizing signals of through a set of computer software.Results:HLA-I alleles were successfully typed in 30 clinical samples .This medium-distinguishing probes were able to discern 57 HLA-I alleles accurately.Conclusion:DNA typing of HLA-I by chip has been proven to be a high-resolution and high-specific method. It is able to check out the new alleles that can not be distinguished by other methods with the same resolution., and it is more intuitional and more suitable for clinical application .

Objective To discuss the value of medium resolution typing method for HLA-B antigens of Northern Chinese by DNA chip technique.Methods The chip was made with specific medium distinguish typing probes designed according to gene frequency of HLA-B alleles from Northern Chinese.Unsymmetrical PCR was used to amplify HLA-B exon 2 and 3, then labeled PCR products and hybridize with probes on the chip.Certified the typing of HLA-B by analysed and scanned the signals of hybridize through a set of computer software.Results HLA-B alleles were successfully typed in 30 clinical samples. This medium distinguish probes were able to discern 42 HLA-B alleles from the scope of HLA-B 7~83. Using it we can distinguish B14,73 and 82 three new alleles contrast to PCR-SSP methods.Conclusions DNA typing of HLA-B by chip was proven to be a high-resolution and high-specificity method. It is able to check out the multitudinous samples in one DNA chip and it is more suitable for clinical application.

Objective To investigate whether toll like receptor ( TLR) signaling pathways can in-crease the expression of IL-17 R in neuralglial cells , and if they can whether the increased IL-17 R is func-tional.Methods Experimental autoimmune encephalomyelitis (EAE) was induced in B6 mice by immuni-zation with an emulsion of myelin oligodendrocyte glycoprotein 35-55 ( MOG35-55 ) in complete Freund's adju-vant (CFA).The expression of Il17ra and Il17rc in the brains and spinal cords of mice with EAE were de-tected by real-time PCR.Luxol fast blue ( LFB) staining was performed to the spinal cord sections to detect tissue demyelination.Immunohistological staining against IL-17RA and CD3 were undertook to visualize IL-17RA+and CD3 + cells.Same approaches were also applied to immunized Rag1 -/- mice to figure out whether T cells infiltration is necessary for increasing IL-17RA expression in the central nervous system ( CNS) .Then B6 mice were immunized with incomplete Freund′s adjuvant ( IFA) plus different TLRs ago-nists to measure the expression of Il17ra in the brains and spinal cords by qPCR .The purified astrocytes , microglia and oligodendrocytes isolated from neonatal mice brains were cultured in vitro for two weeks , and then treated with different TLRs agonists .The expression of Il17ra at mRNA and protein levels in the cells were determined by qPCR and Western blot respectively .The astrocytes were treated with IL-17A and LPS individually or in the combination to detect the level of CCL 2, CXCL8 and IP-10 in the supernatant by ELISA.Results B6 mice with induced EAE showed significantly increased Il17ra expressions in the brain and spinal cord , which was also detected in immunized Rag1 -/-mice.Although no spinal cord demyeliza-tion and CD3 cells infiltration were detected in Rag1 -/-mice, significantly increased number of IL-17RA positive cells could still be visualized .In vivo TLRs agonist participated immunization and in vitro treatment of purified neuroglial cells demonstrated that TLRs agonists could directly evoke IL -17RA expression in the CNS or cultured astrocytes , microglia and oligodendrocytes with high efficiency .Both IL-17 A and LPS could stimulate astrocytes to secrete CCL2, CXCL8 and IP-10, however, a combined use of IL-17A and LPS fur-ther augmented the production of these chemokines to a large extend .Conclusion Taken together , we con-cluded that TLRs agonists could directly stimulate neuroglial cells to express IL -17RA which functionally re-spond to IL-17A by secreting chemokines .

To study the transactivation of a novel thyroid hormone receptor isoform, TR?. pcDNA3. 1-TR? and pGL3-Promoter/thyroid hormone responsive element were co-transfected into COS-7 cells. The expression of reporter gene was detected. It could be increased up to 45 times by T3. TR? and showed the characteristics of a transcription factor.

Objective To compare the results and safety between video-assisted thoracoscopic surgery ( VATS ) and conventional radical operation in patients with stage Ⅰ , Ⅱ esophageal cancer. Methods Retrospectively reviewed 43 patients with stage Ⅰ , Ⅱ esophageal cancer,underwent either VATS radical operation (VATS group,16 cases) or conventional radical operation (control group,27 cases ) from September 2007 to September 2009. Patient's operative characteristics and postoperative courses were compared between two groups. Results In VATS group the operation time was ( 115.6 ± 48.0) min,the peri-operative blood loss was ( 131 ± 71 ) ml,the first postoperative day chest lead quantity was (331 ± 170)ml, the time of postoperative chest tube was (7.25 ± 2.35) d,the postoperative 36 h visual analogue scale (VAS) was (3.4 ± 1.2) scores,the postoperative drainage of chest was ( 1281 ± 534) ml,the 72 h postoperative locomotor activity of right upper extremity was (5.1 ± 1.5) cm. While in control group was ( 145.6 ± 20.6)min, (292 ± 111 ) ml, (494 ± 194) ml, ( 10.00 ± 2.79 )d, (7.3 ± 1.4) scores, ( 1780 ± 731 ) ml, ( 15.6 ± 3.1 )cm respectively (P ＜ 0.01 or ＜ 0.05 ). The lymph node dissection number,the total cost of hospital between were no statistically significant differences in two groups (P ＞0.05). Conclusion Comparing with conventional radical operation, VATS radical operation for patients with stage Ⅰ , Ⅱ esophageal cancer appears to be as effective but less morbid.

Objective:To identify the immune characteristics elicited by immunogen prepared in two different ways against a 36AA-peptide. Methods：Rabbits were immunized by a 36-AA-hapten coupled with carrier proteins or recombinant fusion protein separately. The kinetics and specificities of antibody responses were compared. Results：After about 3 months of primary immunity, the antisera elicited by two kinds of immunogen reached the peak titer. The highest titer elicited by the immunogen prepared with coupled-hapten method reached 1∶12 000, while the highest titer elicited by the immunogen prepared with gene engineering method was 1∶6 000. But the immune response produced by the recombinant fusion protein was more long-lasting. Conclusion：Both kinds of immunogen can successfully elicit 36-AA specific antibody response in rabbits, which have different immune characteristics. To couple the hapten with vector proteins is a quicker and more effective way to prepare the immunogen.

In this study, a Specific Pathogen-Free (SPF) Bama miniature pig herd was established.The standards for breeding experimental pigs, genetic test and microbiological quality control had been drafted.The local standards of Heilongjiang province SPF technical specifications for microbiological monitoring of pigs had been formulated.The genetic and microbiological quality criterion had been used to control of the SPF pig herd.Classical swine fever virus (CSFV) and highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV) infection models of Bama miniature pigs were established, especially the infection miniature pig model of PRRSV was used to evaluated commercially vaccine.The comparison of the cytokine homology between Bama miniature pigs and domestic pigs showed that the Bama miniature pigs can be used to instead of domestic pigs as an ideal experiental animal.At present the SPF Bama miniature pig colonies have been widely used to study the mechanism of prevention and pathogenic in the classical swine fever virus, porcine reproductive and respiratory syndrome virus and porcine transmissible gastroenteritis virus and porcine circovirus and so on.The completion of the project solved the bottleneck problem of the experimental pig usage which provided a new ideal experimental animal for animal disease and life science research.

Objective    To summarize the clinical experience of surgical resection of synchronous multiple ground-glass nodules (SMGN), and explore the individualized diagnosis and treatment strategy of SMGN. Methods    Clinical data of 84 patients with SMGN who underwent thoracic surgery in Anhui Chest Hospital from July 2016 to August 2018 were analyzed retrospectively, including 18 males (21.4%) and 66 females (78.6%), aged 32-80 (55.6±10.3) years. The results of operation and the information of GGNs were analyzed. Results    Except for 1 patient who was converted to thoracotomy due to extensive dense adhesion of thoracic, other patients underwent video-assisted thoracoscopic surgery successfully. All patients recuperated successfully after operation, without severe perioperative complications or death. Finally, 79 patients were diagnosed as malignant tumors (94.0%), and 5 patients of benign lesions (6.0%). A total of 240 GGNs were removed, among which there were 168 pGGNs, including 68 benign lesions (40.5%) and 100 malignant tumors (59.5%), and 72 mGGNs, including 2 benign nodules (2.8%) and 70 malignant tumors (97.2%). Nodules diameter (P<0.001), consolidation/maximum diameter of nodule ratio (P<0.001), vacuole sign (P<0.001), air bronchograms sign (P=0.001), spine-like process (P=0.001), pleural indentation sign (P<0.001), lobulation sign (P<0.001), and vascular convergence (P=0.002) were correlated with malignant tumor. Conclusion    Analysis of the imaging features of GGNs by thin-section CT scan and three-dimensional reconstruction is of great value in predicting the benign and malignant nodules, which can guide the surgical decision-making and preoperative planning. Through reasonable preoperative planning and following certain principles, simultaneous surgical treatment for SMGN is safe and feasible.