Rheumatoid arthritis ( RA) was a common autoimmune disease characterized with main clinical manifestations of chronic inflammation in joints.Anti-carbamylated proteins(anti-CarP)antibody, as a promising bio-marker, played a crucial role in the diagnosis of RA patients and the screening of pre-RA patients .In this article , recent progress on the study of anti-CarP antibody and RA related diseases has been reviewed , which was expected to provide new references for the diagnosis and treatment of related clinical diseases .

Citrullination is a post-translational modification of arginine caused by peptidyl arginine deiminase in the presence of calcium ions.Up-regulation of citrullination plays a significant role in the progress of numerous disorders,and it has important physiologic and pathological significance.High-throughput and quantitative methods with high specificity and sensitivity are still needed to identify the citrullinated sites in tissue cells.With the development of chemical derivatization and mass spectrometry, new rapid and accurate proteomics technology still has a good prospect of development and application.This article aimed to provide new ideas and directions to identify the citrullinated sites in the complex biological samples by introducing strategies available to identify citrullinated peptides, such as color development reagent analysis, immunodetection and mass spectrometry.

Objective: To observe the effects of recombinant adenovirus TRAIL (AdS-TRAIL & Ad5F35-TRAIL) on apoptosis of non-small cell lung (NSCLC) cells, so as to assess the value of Ad-TRAIL in gene therapy of NSCLC. Meth-ods: CAR and CD46 expression levels in lung cancer cell lines (A549, Z793, QG56 and NCI-H520) and the primary lung cancer cells from samples of 10 NSCLC patients were assayed by flow cytometry analysis. The lung cancer cell lines and primary lung cancer cells were infected with Ad5-TRAIL & Ad5F35-TRAIL adenoviral vectors at MOI 10 or 50, re-spectively; the percentage of apoptosis cells labeled by Annexin V-FITC in different cells were measured by flow cytometry 48 h after transfection. Results: The expression of CD46 were higher than that of CAR in all the lung cancer lines (A549, Z793, QG56 and NCI-H520) and the primary lung cancer cells. Significant apoptosis was observed in Z793 and QG56 cells transfected with Ad5-TRAIL or Ad5F35-TRAIL at MOI 10, with the apoptosis rate being (1.76±2.10)% (Ad5-TRAIL), (15.96±2.89) % (Ad5F35-THAIL) and (6.05±1.58) % (Ad5-TRAIL), (10.11±1.26) % (Ad5F35-TRAIL), respectively, compared to no adenovirus-transfected cells ([2.33±0.37] % and [5.95±1.89]%, respectively, P < 0.05). Less than 10% of apoptosis cells were detected in NCI-H520 cells transfected with Ad5- or Ad5F35-TRAIL at MOI 50 ([12.89±3.2] % for AdS-TRAIL and [9.08±1.35]% for Ad5F35-TRAIL, respectively) compared to no adenovirus-transfected cells ([7.04±2.17] %, P > 0.05). Moreover, apoptosis induced by Ad5- or Ad5F35-TRAIL transfection in A549 cells was not detected both at MOI 10 and 50. About half of the primary lung cancer cells from 10 patients induced apoptosis after transfected with Ad5-TRAIL or Ad5F35-TRAIL vector. A higher percentage of apoptotic cells were found in Ad5F35-TRAIL group than those in Ad5-TRAIL and control groups. Conclusion: Ad5-TRAIL can induce apoptosis of NSCLC cells in vitro, and Ad5F35-TRAIL is more potent than Ad5-TRAIL, so Ad5F35-TRAIL is more suitable for gene therapy of NSCLC.

Objectives: By observing and evaluating the clinical curative effect systematically of electrical acupuncture (EA) on Tianshu (ST 25) on diarrhea-predominant irritable bowel syndrome (D-IBS), to make it benefit for the application and spread of EA on Tianshu (ST 25)on D-IBS. Methods: 103 D-IBS matched the involved standards were allocated into treatment group (EA on ST 25, n=53) and control group (EA on Daheng, SP 15, n=50) in randomized,controlled and single-blind ways. Curative effect differences were analyzed by Ridit analysis.Results: The incidence of diarrhea, abdominal pain, abdominal distention or abdominal discomfort, borborygmus, defecation urgency and defecation incompletion feeling were 100%,89.3%, 71.8%, 74.8%, 83.5% and 78.6% respectively in D-IBS. Generally, after treatment, 11cases got excellent results, 34 cases improved and 8 failed in treatment group, totally effective rate was 84.9%; while in control group, 4 cases got excellent effects, 26 improved and 20 failed,totally the effective rate was 60.0%. Conclusion: Compared with control group, EA on Tianshu (ST 25) has a better curative effect on D-IBS.

Objective Rheumatoid arthritis (RA) is a typical type Ⅲ hypersensitivity with a large number of immune complexes (IC) and complement deposits in the synovial tissue , but its specific pathogenesis is not yet clear.This article was to explore the expression of the antigenic profile of serum ICs in RA.Methods ICs were isolated from the serum of 55 patients with RA (41 cases of anti-CCP antibody [+] and 14 cases of anti-CCP antibody [-]), 41 with systemic lupus erythematosus (SLE), and another 41 healthy controls by polyethylene glycol (PEG) precipitation, separated by immunoprecipitation, digested with trypsin in gel, and then subjected to mass spectrometry for identification.The levels of total proteins were compared among different groups using Vennny 2.1.0.The protein expression was considered to be up-regulated when the total protein level of the RA group was >2 times and down-regulated when it was <0.5 times that of the control.Further functional analysis was performed on the differential proteins in RA using the STRING software.Results Totally, 277 proteins were identified in the serum ICs of the RA patients, including 162 in the anti-CCP (+) and 248 in the anti-CCP (-) RA group.Compared with the SLE and healthy control groups, only 129 proteins were found in the RA patients, including 38 in the anti-CCP (+), 109 in the anti-CCP (-) RA group, and 18 in both the two groups.Among the proteins identified in the RA patients and healthy controls, 2 and 11 were up-regulated while 17 and 21 down-regulated in the anti-CCP (+) and anti-CCP (-) RA group, respectively.Conclusion More differentially expressed proteins were identified in the anti-CCP (-) than in the anti-CCP (+) RA patients.The identification of differentially expressed proteins provides a new idea and direction for the investigation of the pathogenesis and new biomarkers of RA.

Objective The inhibitor of differentiation 3 (Id3) is an important transcriptional regulation factor, which participates in tumorigenesis, cell proliferation, and cell apoptosis.β-catenin, as a central molecule of the Wnt signaling pathway, is critical for tumor development.This study aimed to evaluate the expressions of these two molecules and the regulatory effect of Id3 on β-catenin in different tumor cells.Methods Total RNA was extracted using the Trizol Reagent.The relative mRNA expression levels of Id3 and β-catenin in tumor cells were detected by quantitative real-timePCR(qRT-PCR).The recombinant eukaryotic expression vector pEGFP/Id3 with the human Id3 gene was transfected into A549, A549/ DDP and SW-480 cells using the non-liposome-mediated method.The protein expressions of Id3 and β-catenin were determined by Western blot.Results The expression of Id3 was significantly lower in the colorectal cancer cell lines SW-480 and HT-29 than in A549 and other tumor cells (P<0.05), but remarkably higher in nasopharyngeal carcinoma CNE and 5-8F cells than in other tumor cells (P<0.05).The expression of β-catenin was the highest in SW-480in comparison withother malignant tumor cells(P<0.05), and the second highest was in gastric cancer AGS and colorectal cancer HT-29 cell lines, but low in H446, A549, SPC-A-1, A549/DDP, and SK-MES-1 cell lines and extremely low or almost absent in CNE and 5-8F cells (P<0.05).After transfected with pEGFP/Id3, the cells showed a decreased volume, wrinkled membrane and absent refraction under the fluorescence microscope, which, however, were not observed in most of the cells transfected with the empty vector pEGFP.Compared with the control, the Id3/pEGFP group showed remarkably increased expressions of Id3 mRNA in the A549, A549/DDI, and SW-480 cells (1.24±0.12 vs 193.12±2.80, 1.09±0.11 vs 188.30±2.60, and 0.92±0.29 vs 19.08±0.59, P<0.01), and the expression of β-catenin was significantly down-regulated in the transfected SW-480 cells with an overexpression of Id3 (0.98±0.05 vs 0.32±0.03, P<0.01), but exhibited no statistically significant differences from those in the transfected A549 and A549/DDP cells (0.98±0.07 vs 1.04±0.08 and 0.98±0.05 vs 0.32±0.03, P>0.05).Western blot showed the same results.Conclusion The expression levels of Id3 and β-catenin vary in different tumor cell lines.Anabnormally high level of β-catenin is an important risk factor for colorectal cancer, and the down-regulatedexpression of β-catenin after eogenous transfection of Id3 may provide some new ideas for target gene therapies of colorectal cancer.

Objective To scan protein expression profile of immune complexes (ICs) derived from the synovial tissue of the patients with rheumatoid arthritis (RA) based on liquid chromatography-tandem mass spectrometry (LC-MS).Methods The samples of synovial fluid were obtained from knee joints of the patients with RA and osteoarthritis (OA) used as control during therapeutic arthrocentesis in knee jiont at the Department of Orthopedics of Jinling Hospital,School of Medicine,Nanjing University.The protein expression profile of ICs was identified by enrichment strategy based on immunoprecipitation and LC-MS analysis.The value of fraction of total (FOT) was used to estimate protein abundance and screen the up-and down-regulated proteins.The function enrichment,interaction network and signal pathway of differential proteins were analyzed using softwares David and String.Results A total of 511 and 526 protein spots in ICs of RA and OA patients were identified respectively.Among them,170 proteins existed only in RA group.45 and 85 proteins in RA group were statistically up-and down-expressed compared with controls.Conclusion HSP90AA1,HSP70,HLAG,Thioredoxin,Annexin A2 and vitronectin may be involved in the pathogenesis of RA through different paths and possible to become promising diagnostic indicators or new therapeutic targets for RA.