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.

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 .

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: 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 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.

Objective To summarize the treatment experience and strategies of patients with acute type A aortic dissection involving coronary arteries in order to improve the efficacy of such patients.Methods Between March 2013 and April 2016,we recruited 37 patients with coronary involvement caused by acute type A aortic dissection,26 men,11 women;mean age (49.7 ± 10.4) years.All procedures were done on an emergency basis within 24 hours after the patient's arrival.Results Acute type A aortic dissection with coronary involvement is a more complex operation associated with high in-hospital mortality(18.9％,7/37) and low short-term survival(64.9％,13/37).There were 9 patients underwent coronary artery bypass graft after completion of the root procedure because of ventricular wall motion abnormality(7 patients) and new ST-segment elevation (2 patients) during weaning from cardiopulmonary bypass.Four of them were survival during follow-up due to the success from rescue coronary artery bypass graft.Conclusion Acute type A aortic dissection with coronary involvement is associated with high in-hospital mortality and low short-term survival.If patients suffered abnormal ventricular wall motion or new ST-segment elevation during weaning from cardiopulmonary bypass,rescue coronary artery bypass graft is essential to salvage these critically ill patients.

Objective:To investigate the prevalence and independent risk factors of non-alcoholic fatty liver disease (NAFLD) and advanced chronic liver disease among the type 2 diabetes mellitus (T2DM) population in the Shenyang community, so as to provide evidence for the prevention and control of T2DM combined with NAFLD.Methods:This cross-sectional study was conducted in July 2021. 644 T2DM cases from 13 communities in Heping District, Shenyang City were selected. All the surveyed subjects underwent physical examination (measurements of height, body mass index, neck circumference, waist circumference, abdominal circumference, hip circumference, and blood pressure), infection screening (excluding hepatitis B and C, AIDS, and syphilis), random fingertip blood glucose, controlled attenuation parameter (CAP), and liver stiffness measurement (LSM). The study subjects were divided into the non-advanced chronic liver disease group and the advanced chronic liver disease group according to whether the LSM value was greater than 10 kPa. Cirrhotic portal hypertension development was indicated in patients with LSM ≥ 15 kPa. The comparison of multiple mean values among the sample groups was performed by analysis of variance when the normal distribution was met.Results:In the T2DM community population, there were 401 cases (62.27%) combined with NAFLD, 63 cases (9.78%) combined with advanced chronic liver disease, and 14 cases (2.17%) combined with portal hypertension. There were 581 cases in the non-advanced chronic liver disease group and 63 cases (9.78%) in the advanced chronic liver disease group (LSM ≥10 kPa), including 49 cases (7.61%) with 10 kPa≤LSM<15 kPa, 11 cases (1.71%) with 15 kPa ≤LSM<25 kPa, and 3 cases (0.47%) with LSM ≥ 25 kPa. Age, body mass, body mass index, neck circumference, waist circumference, hip circumference, waist-to-height ratio, systolic blood pressure, and CAP were all statistically different between the non-advanced chronic liver disease group and the advanced chronic liver disease group ( F=-1.983,-2.598,-4.091,-2.062,-3.909, -4.581,-4.295,-2.474, and -5.191, respectively; P<0.05). There was a statistically significant difference in terms of whether or not there was combined cerebrovascular disease (2=4.632, P=0.031); however, there were no statistically significant differences in terms of lifestyle, diabetes complications, and other complications ( P>0.05). Conclusion:Patients with T2DM have a higher prevalence of NAFLD (62.27%) than those with advanced chronic liver disease (9.78%). 2.17% of T2DM cases in the community may not have had early diagnosis and early intervention, and they might have been combined with cirrhotic portal hypertension. So, the management of these patients should be strengthened.

Objective:To evaluate the clinical effects of anterolateral thigh free flap with fascia lata in the repair of dura mater defect after resection of head squamous cell carcinoma.Methods:From June 2016 to June 2018, Xijing Hospital of Air Force Medical University applied the free transplantation of anterolateral thigh flap with fascia lata to repair the dura mater defect of 12 patients with head squamous cell carcinoma, including 9 males and 3 females, aged from 35 to 74 years. The size of scalp soft tissue defects in patients after carcinoma resection ranged from 12 cm×10 cm to 24 cm×21 cm, and the size of dura mater defect of patients ranged from 7 cm×6 cm to 16 cm×14 cm. The size of flap of patients ranged from 14 cm×12 cm to 27 cm×24 cm, and the size of fascia lata ranged from 8 cm×7 cm to 17 cm×15 cm. The superficial temporal artery and middle temporal vein were connected by end to end anastomosis with the first musculocutaneous perforating branch of the descending branch of lateral femoral artery and its accompanying vein. The flap donor area was transplanted with autologous split-thickness skin graft from trunk and fixed with packing. Postoperative survival of flaps and skin grafts was observed. The patients were followed up regularly. The cranial magnetic resonance imaging was performed to observe the recurrence of intracranial tumors and dural integrity, shape of the flap and whether the donor site region was left with significant dysfunction were observed.Results:All the flaps and skin grafts survived well in 12 patients after surgery. Ten patients had primary healing at the edge of the flap suture; 2 patients had local sinus tract formation at the suture site of flap, with a small amount of cerebrospinal fluid leakage, and were recovered after outpatient dressing change. The patients were followed up for 10 to 36 months, and 3 patients with tumors involving in the dura mater sagittal sinus region had postoperative intracranial tumor recurrence. The tumor was resected again. All the patients had good dural integrity. The flaps of all patients were in good shape, and no obvious dysfunction remained in the flap donor site.Conclusions:Free transplantation of anterolateral thigh flap with fascia lata is an effective and reliable method to repair the dura mater defect following head squamous cell carcinoma resection. It can repair the scalp and dura mater defects caused by the invasion of squamous cell carcinoma and provide possibilities for skull reconstruction.