1.Clinical application of anti-carbamylated protein antibody
Journal of Medical Postgraduates 2016;29(4):421-426
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 .
2.Methods for identification of citrullinated proteins in tissue cells
Journal of Medical Postgraduates 2017;30(4):421-426
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.
3.Clinical Research of Irritable Bowel Syndrome Treated by Electroacupuncture on Tianshu(ST 25)
Huirong LIU ; Huangan WU ; Xiaolong WANG ; Chuantao ZUO ; Yihui GUAN ; Jianqiao FANG
Journal of Acupuncture and Tuina Science 2007;5(2):91-94
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.
4.CAR- or CD46-dependent TRAIL-adenoviral vector induced apoptosis in lung cancer cells
Lingdi MA ; Yong WANG ; Cheng NI ; Shizhong WANG ; Yongyi BAO ; Naifu GUAN ; Ke ZHANG ; Xiaolong FAN
Chinese Journal of Cancer Biotherapy 2009;16(6):577-582
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.
5.Differentially expressed proteins in serum immune complexes of rheumatoid arthritis: Analysis by mass spectrometry
Xiaolong GUAN ; Haiyong WANG ; Ying ZHOU ; Yuzhong XU ; Ang LI ; Shusheng SONG ; Xiaojun LI ; Wei YU
Journal of Medical Postgraduates 2017;30(5):495-501
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.
6.Regulatory effect of overexpressed Id3 on the expression of β-catenin in different tumor cells
Qinfei ZHAO ; Fangfang CHEN ; Yuzhong XU ; Haiyong WANG ; Xiaolong GUAN ; Ning SUN ; Ang LI ; Shusheng SONG ; Wei YU ; Xiaojun LI
Journal of Medical Postgraduates 2017;30(5):464-469
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.
7.Differential proteomic analysis for immune complexes deposited in synovial tissues of RA patients using liquid chromatography-tandem mass spectrometry
Haiyong WANG ; Xiaolong GUAN ; Qinfei ZHAO ; Yuzhong XU ; Yu ZHANG ; Shusheng SONG ; Ang LI ; Xing LV ; Ying ZHOU ; Wei YU ; Jianning ZHAO ; Xiaojun LI
Chinese Journal of Clinical Laboratory Science 2017;35(1):15-20
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.
8.Surgical treatment for 37 patients with acute type A aortic dissection involving coronary arteries
Ming GONG ; Xinliang GUAN ; Xiaolong WANG ; Yuyong LIU ; Haiyang LI ; Wenjian JIANG ; Jiachen LI ; Yang LIU ; Hongjia ZHANG
Chinese Journal of Thoracic and Cardiovascular Surgery 2018;34(8):465-468
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.
9.Observation of neuropsychological development status in children after surgical treatment of congenital heart diseases
Shuantong LIN ; Ying LIANG ; Xiaolong WANG ; Xinguang WEI ; Jingxin YAO ; Dianyuan LI ; Hao ZHANG ; Cun LONG ; Fu-Qing JIANG ; Yulong GUAN
Chinese Journal of Thoracic and Cardiovascular Surgery 2018;34(11):683-687
Objective To observe neuropsychological development status in children after surgical treatment of congenital heart diseases(CHDs)and analyze the risk factors. Methods 89 children who received outpatient review in Fuwai Hospital from September 2015 to March 2016 after surgical treatment of CHDs were recruited in this study and 90 normal children were recruited as the control group. The children with CHDs were divided into simple CHDs group(RACHS- 1 score≤2)and com-plex CHDs group(RACHS- 1 score≥3)according to RACHS- 1 classification. Neuropsychological development status was meas-ured according to pediatric-psychological mental test scale developed by Capital institute of pediatrics,Beijing and statistical a-nalysis was compared. Results The measurements of neuropsychological development showed the normal children behaved better than the children with CHDs(P < 0. 05). The simple CHDs group achieved better distribution of development quotient than complex CHDs group(P = 0. 032)and there was no difference between the normal control group and simple CHDs group (P = 0. 420). Multivariate regression analysis indicated that younger age at cardiac surgery,lower preoperative blood urea ni-trogen(BUN),higher preoperative creatinine(Cr)and prolonged duration of cardiopulmonary bypass(CPB)accounted for low-er scores in the test scale(P < 0. 05). Conclusion Distinct neuropsychological difficulties could be present especially in chil-dren with complex CHDs. Younger age at cardiac surgery,preoperative BUN,Cr and CPB duration were perioperative factors that were associated with long-time neuropsychological development.
10.TMEM16A inhibits angiotensin II-induced basilar artery smooth muscle cell migration in a WNK1-dependent manner.
Huaqing ZHENG ; Xiaolong LI ; Xin ZENG ; Chengcui HUANG ; Mingming MA ; Xiaofei LV ; Yajuan ZHANG ; Lu SUN ; Guanlei WANG ; Yanhua DU ; Yongyuan GUAN
Acta Pharmaceutica Sinica B 2021;11(12):3994-4007
Vascular smooth muscle cell (VSMC) migration plays a critical role in the pathogenesis of many cardiovascular diseases. We recently showed that TMEM16A is involved in hypertension-induced cerebrovascular remodeling. However, it is unclear whether this effect is related to the regulation of VSMC migration. Here, we investigated whether and how TMEM16A contributes to migration in basilar artery smooth muscle cells (BASMCs). We observed that AngII increased the migration of cultured BASMCs, which was markedly inhibited by overexpression of TMEM16A. TMEM16A overexpression inhibited AngII-induced RhoA/ROCK2 activation, and myosin light chain phosphatase (MLCP) and myosin light chain (MLC20) phosphorylation. But AngII-induced myosin light chain kinase (MLCK) activation was not affected by TMEM16A. Furthermore, a suppressed activation of integrin