Objective To detect anti-heterogeneous nuclear ribonucleoprotein A2 (hnRNP A2)/RA33 antibody by ELISA with the purified recombinant hnRNP A2 antigen. Methods The serum of 179 patients with RA, 141 patients with SLE, 97 patients with other diffused rheumatic diseases, 30 patients with seronegative spondyloarthropathies, 10 patients with osteoarthritis, 59 patients with arthralgia/arthritis and 40 controls were detected. In addition, clinical characters and laboratory indexes were compared to study the significance of anti-hnRNP A2/RA33 antibody in RA. Results The sensitivity and specificity of anti-hnRNP A2/RA33 antibody in RA were 36.9% and 87.1%. The positive rates of anti-hnRNP A2/RA33 antibody in SLE, other CTD, seronegative spondyloarthropathies and OA were 19.2%, 7.2%, 6.8% and 0. The positive rate of anti-hnRNP A2/RA33 antibody was 43.3% in early RA patients. Conclusion Detection of anti- hnRNP A2/RA33 antibody with purified recombinant hnRNP A2 antigen is a reliable method for early diagnosis of RA.

Objectives To assess the presence of autoantibodies directed against the epitopes of SmB and SmD in systemic lupus erythemotosus(SLE) as well as other different connective tissue diseases(CTDs) and analyze the clinical significance of them.Methods Polypeptides of SmB and SmD were designed with the animo acids sequence by the biologic technical software.The sera from different patients including 102 SLE,153 other CTDs and 54 controls were detected by ELISA with thesynthetic polypeptides.Results The positive percentage of anti-SmB and antiSm-D epitopes were higher in SLE than in other groups (P

Objective To explore the effects of visfatin in glucose metabolism by testing the expression of visfatin in liver of rats in different glucose metabolic statuses .Methods SD rats were randomly divided into five groups :normal control group (NC group) ,diet induce obesity group (DIO group) ,diabetes mellitus group (DM group) ,diabetes controlled by insulin group (INS group)and diabetes controlled by metformin group (MET group) .Tested the data of blood glucose (FPG) ,triglyceride(TG) ,total cholesterol(TC) ,free fat acid(FFA) ,fasting insulin(Fins) .The liver of rats was used to test visfatin ,glucose‐6‐phosphatase(G‐6‐pase) mRNA by RT‐PCR and visfatin ,AMP‐activated protein kinase‐α (AMPKα) ,phosphor‐AMP‐activated protein kinase‐α (p‐AMPKα) protein by Western blot .Results FBG of group DM increased than group NC and DIO (P< 0 .01) ;FBG of group INS and MET decresed than group DM (P < 0 .01) ;HOMA‐IR of group DIO and DM increased than group NC (P < 0 .01) ;HOMA‐IR of group DM increased than group DIO (P< 0 .01) .ISI of group DIO and DM decresed than group NC (P< 0 .01) ;ISI of group DM de‐creased than group DIO(P< 0 .01) .TG of group DIO increased than group NC (P< 0 .01) .TG of group INS and MET decreased than group DM (P< 0 .05) .The level of TG and TC of group DM ,INS and MET increased than group NC (P< 0 .05) .The level of serum FFA of group DM ,INS and MET were significantly higher than group NC (P < 0 .05) ;FFA of group DM increased than group DIO(P< 0 .05) .The expression of visfatin mRNA of group DM increased than group NC and DIO (P< 0 .05) ;visfatin mRNA of group INS and MET decreased than group DM (P< 0 .01) .Group DM ,INS and MET had a significantly higher level of G‐6‐Pase mRNA of than group DIO( P < 0 .05) ;Group MET had a significantly lower level of G‐6‐Pase mRNA of than group DM (P <0 .05) .The expression of visfatin protein of group DM ,INS and MET increased than group NC ( P < 0 .05) .The expression of AMPKα protein of group DM ,INS and MET decresed than group NC(P< 0 .05) ;AMPKα of group DM decresed than group DIO (P<0 .05) .The expression of p‐AMPKα protein of group DIO ,DM ,INS and MET decresed than group NC (P< 0 .01) .Conclusion The ex‐pression of visfatin in liver of SD rats might have something to do with insulin resistance and diabetes .We could′t consider that visfatin can affect the pathway of metformin activated AMPK to decrease blood glucose .

Objective To establish and apply the protein chip to detect eleven autoantibodies profile, and evaluate the authenticity and reliability with ANAs protein chip in clinical autoantibodies profile detection.Methods By comparing the results of IIF and ELISA , validation the sensitivity and specificity of ANAs protein chip in clinical autoantibodies profile detection. The autoantibodies detected were anti-SSA-52,anti-SSA-60,anti-SSB,anti-Sm,anti-RNP,anti-Scl-70,anti-Jo-1,anti-dsDNA,anti-rRNP,anti-centromere antibodies and antinuclear antibodies (ANA). To each autoantibody, we have selected 70 positive and 294 negative samples except the 32 rare samples that contain anti-Jo-1 antibody.Results The sensitivity to all the autoantibodies was 100% except anti-SSA52 and anti-SSB antibodies was 95.7%and 98.6% respectively. The specificity to all the autoanbodies was 100% except anti-SSB, anti-RNP-68, anti-Scl-70, anti-dsDNA, anti-CENP-B and ANA was 98.0%, 98.0%, 99.7%, 99.7%, 99.7% and 98.3% respectively. Conclusions To all the eleven antinuclear autoantibodies , the sensitivity is all above 95.0% and specificity is all above 98.0%, which indicate that there is high concordances between the ANAs protein chip and the methods used in clinical screening and confirmation,and it could meet the requirement of clinical autoantibodies profile detection. The protein chip method is fast, easy for detection with the characteristic of high-throughput,high sensitivity and specificity,it is hence recommended to apply ANAs protein chip to detect autoantibodies profile in clinical detection.

Objective To clone human Sjogren's syndrome antigen A(SSA)for expressing of antigen SSA-52kD and establishing a new clinical detecting method.Methods According to the human SSA-52kD cDNA sequence reported in GenBank,primers of human SSA-52kD cDNA were designed and synthesized.Human SSA-52kD cDNA was amplified from RNA of cultured Hela cell by reverse transcriptase polymerase chain reaction(RT-PCR).The production of amplification was ligated to PET-30a vector and then transformed into the competent bacteria DH5?to construct the recombinant plasmid PET-30a-SSA-52kD.The recombinant plasmid was digested with Bgl Ⅱ and Hind Ⅲ,and positive clones were sequenced.Results The Human SSA-52kD cDNA fragment containing 1447bp was amplified by RT-PCR.Restriction endonuclease mapping using Bgl II and Hind III showed that the target gene was inserted into the recombinant plasmid.The complete coding sequence of Human SSA-52kD was consistent with that of GenBank through DNA sequencing.Conclusions The full length of human SSA-52kD cDNA was successfully cloned and the recombinant plasmid PET-30a-SSA-52kD was constructed.

Objective To evaluate the BALB/c murine infective effects in different concentrations and different aerosol challenge times by Bordetella pertussis.Methods Four experiment groups according to different concentrations and different aerosol challenge times were designed.BALB/c murines were challenged by aerosol way.Group 1: 1010cfu/mL Bordetella pertussis challenge 15 min, group 2: 1010cfu/mL challenge 30 min, group 3: 109cfu/mL challenge 30 min, group 4: 1011cfu/mL challenge 30 min, using the normal saline challenge 30 min as control.At 0d,3d,7d,14d and 21d after challenge, the WBCs of all groups were measured and lung tissues were homogenized to calculate the bordetella pertussis clone in lung.Results After 3 days of challenge, WBCs in all groups were slightly increased.The WBCs of group 1, group 2, group 3 and group 4 were significantly increased after 7 days, with the average numbers of 8.52×109 per/L, 1.74×1010per/L, 1.15×1010per/L and 5×1010per/L, respectively.After 14 days, they were 1.77×1010per/L, 1.67×1010per/L, 1.27×1010per/L and 3.84×1010per/L respectively.WBCs in all groups were dramatically declined after 21 days.The WBC of negative control group had no obvious change during the whole process with the stable number of 3.4~7.0×109per/L.Bordetella pertussis were detected in lung of all experimental groups in each sampling point.The CFU in lung wase at peak at 7d or 14d after challenge, which was obviously decreased at 21d.Conclusion This aerosol challenge method can establish a bordetella pertussis infection mouse model successfully.

Objective:To construct and prepare recombinant virus strains chimeric with human metapneumovirus (HMPV) antigenic epitopes.Methods:Recombinant influenza virus vectors which chimeric with different HMPV antigenic epitopes were rescued by reverse genetics using eight-plasmid system. The recombinant influenza virus strain used the internal genes of A/PR/8/34 (PB1, PB2, PA, NP, NS, M, HA, and NA) as a backbone, with concomitant genetic modifications to insert the B-cell epitopes of HMPV into the HA gene, and the CTL+ Th cell epitopes of HMPV into the NA gene. Preparation of recombinant influenza virus strains using reverse genetics in a " 7+ 1" model. The recombinant virus strains were evaluated by measuring hemagglutinin (HA) titers, half tissue culture infectious dose (TCID 50) and growth curves. Sequencing analysis was conducted to verify whether the rescued viruses carried the chimeric HMPV epitopes. Results:The epitopes of HMPV were inserted into the influenza virus genome and two recombinant influenza virus strains were rescued successfully, named as FLU/HMPV/B and FLU/HMPV/CTL+ Th. HA titers of the recombinant strains were both 2 7, their TCID 50 were 10 5.2/ml and 10 5.0/ml, respectively. After cultured for three passages in chick embryo, these two recombinant strains could proliferate steadily. Whole genome sequencing verified that the FLU/HMPV/B carried the B-cell epitopes of HMPV, the FLU/HMPV/CTL+ Th carried the CTL and Th cell epitopes of HMPV. Growth curve tests also verified that the recombinant strains could proliferate steadily in eggs. Conclusions:Two recombinant influenza virus vector strains carrying the B cell, CTL and Th epitopes of HMPV were rescued successfully. The result of the recombinant virus strains in terms of growth characteristics as well as genetic stability indicate that they meet the requirements for proceeding to the next step of animal experiments. The immunogenicity and immunoprotective effect will be further evaluated by mouse experiments. Ultimately new ideas for the realization of " one vaccine for two uses" or " one vaccine formultiple uses", as well as a new strategy for the development of HMPV vaccine will be proposed.

OBJECTIVE: To investigate the effects of serum from the obesity patients and obesity patients with Diabetic mellitus on toll-like receptor 4/Nuclear factor -κB p65 (TLR/NF-κB) pathway in human THP-1 monocytes and to explore the inflammatory immune response in obesity. METHODS: Peripheral serum was isolated from healthy volunteers (the control group), the obesity patients (Ob group) and the obesity patients with diabetic mellitus (the Ob with DM group), respectively, 20 in each group. THP-1 monocytes were incubated with the serum for 48 h. The monocytes and culture supernatant were collected. The phosphorylation level of NF-κB p65 protein in THP-1 monocytes was evaluated by Western blot as well as immunofluorescence assay. The TLR4 mRNA expression was evaluated by RT-PCR. ELISA was used to measure the monocyte chemotactic protein-1 (MCP-1) levels in the culture supernatant. RESULTS: In the presence of serum, the obesity group and the obesity with diabetic mellitus group showed the up-regulated phosphorylation level of NF-κB p65 protein and TLR4 mRNA expression in THP-1 monocytes compared with the healthy control group (both P<0.05), and the MCP-1 levels in the obesity patients were up-regulated significantly compared with the healthy control group [healthy control group (26.4 ± 3.9) pg/mL, Ob group (45.8 ± 10.0) pg/mL, Ob with DM group (58.0 ± 15.3) pg/mL; P<0.05]. These parameters were further up-regulated in the obesity patients with diabetic mellitus patients. CONCLUSION The serum from the obesity patients or the obesity patients with diabetes can induce monocyte dysfunction, which might be related to the activation of TLR4/NF-κB signaling pathway.
Cell Line ; Chemokine CCL2 ; Diabetes Mellitus ; blood ; Humans ; Monocytes ; cytology ; metabolism ; Obesity ; blood ; Phosphorylation ; Serum ; Signal Transduction ; Toll-Like Receptor 4 ; metabolism ; Transcription Factor RelA ; metabolism ; Up-Regulation

Objective To establish an intestinal organoid-based assay to investigate the radiation mitigation effect of epiregulin in vitro. Methods Intestinal crypts were released from tissue incubated with EDTA. Intestinal crypts seeded in 3D matrigel were irradiated at 24 h after plating. The radiation mitigation effect of epiregulin was evaluated by measuring the survival rate, size and budding numbers of the organoid after irradiation, and the basic FGF was used as a positive control of epiregulin. Results Radiation-induced lethality and dose-dependent survival curve of the intestinal organoid were consistent with in vivo data. Treatment with epiregulin (400 ng/ml) at 24 h post-radiation significantly increased survival rate of 8 Gy X-ray irradiated intestinal organoid in comparison with non-treated group [(12.56 ± 1.02)％vs. (4.73 ± 0.38)％, t=12.43,P<0.05]. Conclusions Epiregulin has radiation mitigation effect on intestinal organoid and could serve as a potential medical countermeasure to mitigate gastrointestinal toxicity.

Objective:To investigate the expression of osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) in type 2 diabetic rats with periodontitis.Methods:46 male Wistar rats were randomly divided into the healthy group(n =10),the periodontitis group(n =12),the type 2 diabetes mellitus group(n =12) and the type 2 diabetic periodontitis group(n =12).Animal models were prepared respectively.The expression of OPG and RANKL protein in alveolar bone was detected by immunohistochemistry(IHC).Results:Compared with the healthy group,the expression of OPG in the type 2 diabetes mellitus group,the periodontitis group,the type 2 diabetes mellitus with periodontitis group decreased in turn,however the expression of RANKL increased in turn.The expression of OPG and RANKL had no significant difference between periodontitis group and type 2 diabetes periodontitis group,while there was statistically significant difference among the other groups (P ＜ 0.05).Conclusion:Inflammation may lcad to upregulation of RANKL in osteoclasts and immune cells,and downregulation of OPG in osteoblasts.