1.Study on immunogenicity and arthritogenicity of a synthetic cyclic citrullina-ted peptide
Ensheng CHEN ; Mingzhu CUI ; Xiaofeng ZHAO ; Beijia YU ; Changhong XIAO ; Weiwang GU
Chinese Journal of Immunology 2017;33(1):25-30
Objective:To establish a synthetic cyclic citrullinated peptide induced arthritis model in mice, explore immunogenicity and arthritogenicity of this peptide. Methods: 36 DBA/1 mice were randomly divided into three groups, which were injected the type Ⅱ collagen ( CⅡ, CIA ) emulsion, cyclic citrullinated vimentin peptide ( CCit-Vim, CCV-IA ) emulsion, cyclic citrullinated vimentin peptide conjugated KLH ( CCit-Vim+KLH,CCV+K-IA) emulsion on day 0 and 21,respectively. Using arthritis index( AI) ,paw swelling to evaluate the incidence of arthritis;ELISA tested serum anti-CCit-Vim antibody,anti-CⅡantibody,anti-CCP antibody and TNF-α, IIF detected AKA;Histopathology of the ankle joint was obsearved. Results: There were three mice appeared arthritis in CCV+K-IA,the incidence rate of 25%,but arthritis occurs later time,short duration,and the incidence and extent of arthritis were lower than the CIA. CCV-IA no arthritis performance. CCV+K-IA produce anti-CCit-Vim antibody were significantly higher than those in CIA (1. 32±0. 59 vs 0. 78±0. 27,P=0. 031). While Anti-CCP antibody of CCV+K-IA were significantly lower than CIA (54. 73±7. 33 vs 64. 37±9. 91,P=0. 007). The anti-CⅡ antibody in CCV+K-IA and CCV-IA were lower than the CIA(15. 73±2. 10, 16. 71±3. 03 vs 19. 50±2. 36,P<0. 05). The TNF-α produced by CCV+K-IA and CIA were both significantly higher than the CCV-IA (645. 61±35. 26,618. 98±53. 32 vs 533. 63±79. 49,P<0. 05). The AKA positive rate of CCV+K-IA is 50% (6/12),significantly higher than CCV-IA 25% ( 3/12 ) and CIA 16. 67% ( 2/12 ) . Histopathology of the ankle showed that the CCV-IA and CCV+K-IA have a mild synovial hyperplasia,no obvious synovial pannus formation and inflammatory cell infiltration. Conclusion:The cyclic citrul-linated peptide conjugated KLH not only has stronger immunogenicity but also has arthritogenicity. It induced a higher positive rate of AKA than CⅡ.
2.Short-term curative effect of superior rectal artery preserving laparoscopic resection of sigmoid colon carcinoma
Xiaolong TANG ; Hui QU ; Qingsi HE ; Guorui SUN ; Hao ZHANG ; Ensheng ZHAO
Chinese Journal of General Surgery 2018;33(1):30-33
Objective To study the operational techniques and feasibility of superior rectal artery preserving laparoscopic resection of sigmoid colon carcinoma.Methods From Jan 2015 to Nov 2016,85 patients with sigmoid colon carcinoma were divided into artery preserving group (27 cases) and traditional surgical group (58 cases).Results The mean operation time was (283 ± 51) min,the mean lymph node dissection was (15 ± 8) and the mean blood loss was (62 ± 17) ml in the artery preserving group.The mean operation time was (179 ±e63) min,the mean lymph node dissection was (15 ±7) and the mean blood loss was (67 ± 17) ml in the traditional surgery group.The number of resected lymph nodes and blood loss had no statistical significance between these two groups (t =0.058,P >0.05).Longer operating time were observed in the retained vascular group as compared to the traditional surgical group (t =7.530,P < 0.05).There was no anastomotic fistula in the retained vascular group,however,two anastomotic fistula cases occurred in the traditional surgical group (x2 =0.043,P > 0.05).Conclusions Preservation of superior rectal artery was safe and feasible for laparoscopic resection of sigmoid colon carcinoma.
3.Differential expression analysis of absent in melanoma 2-inflammasome pathway associated protein in rheumatoid arthritis and osteoarthritis synovium
Fujuan QIU ; Yong CHEN ; Xiaofeng ZHAO ; Ensheng CHEN ; Fangfang ZUO ; Yi YUAN ; Zixun WU ; Qin SU ; Changhong XIAO
Chinese Journal of Rheumatology 2020;24(6):383-387,C6-1
Objective:To compare the expression levels of absent in melanoma 2 (AIM2) inflammasome pathways in rheumatoid arthritis (RA) and osteoarthritis (OA) synovial specimens.Methods:Synovial tissue samples were collected from 41 RA and 26 OA patients, respectively. Horseradish peroxidase immunohi stochemical staining was used to detect AIM2 inflammasome pathway-related proteins, including AIM2, apoptosis-associated speck-like protein containing a CARD(ASC), caspase-1, and interleukin-1 (IL-1β). A semi-quantitative score (H-score) was performed according to the degree of positiveness. Correlation analysis between H-score results and clinical indicators of erythrocyte sedimentation tate (ESR) and C-reactive protein (CRP) were performed. The H score between RA and OA was analyzed by t test and Spearman correlation analysis were utilized for correlation analysis between H score and ESR and CRP.Results:The H scores of AIM2 protein in RA synovial tissues was (132±7) and (54±8) in OA synovial tissues ( t=7.42, P<0.01). The H scores of ASC protein in RA synovial tissues was (107±9) and (74±6) in OA synovial tissues ( t=2.36, P<0.05). The H scores of caspase-1 protein in RA synovial tissues was (99±5) and (74±10) in OA synovial tissues ( t=2.15, P<0.05). The H scores of IL-1β protein in RA synovial tissues was (118±11) and (76±7) in OA synovial tissues ( t=3.30, P<0.05) . In the correlation analysis, AIM2 was positively correlated with ESR [ r=0.74, P<0.01, 95% CI(0.38, 0.9)], and ASC was positively correlated with ESR [ r=0.5, P<0.05, 95% CI(0.16, 0.74)], IL-1β was positively correlated with ESR [ r=0.62, P<0.05, 95% CI (0.31, 0.81)], and the difference was statistically significant ( P<0.05). At the same time, AIM2 was positively correlated with CRP [ r=0.65, P<0.05, 95% CI(0.25, 0.86)]; ACS was positively correlated with CRP [ r=0.42, P<0.05, 95% CI(0.05, 0.69)]. IL-1β was positively correlated with C-reactive Protein [ r=0.41, P<0.05, 95% CI(0.05, 0.67)] and positively correlated with C-reactive protein, and the difference was statistically significant ( P<0.05). Conclusion::The expression of AIM2 inflammasome pathway-related proteins in RA synovium, including AIM2, ASC, caspase-1, and IL-1β, is higher than that of OA and are positively correlated with disease activity. Activation of AIM2 inflammasome pathway may be associated with the pathogenesis of RA disease activity.
4. decoction induces apoptosis by activating Fas/caspase-8 pathway in rheumatoid arthritis fibroblast-like synoviocytes.
Fan ZHAO ; Jiayu LI ; Qijin LU ; Ensheng CHEN ; Lixia YUAN
Journal of Southern Medical University 2020;40(8):1119-1126
OBJECTIVE:
To explore the effect of decoction (DGNTD) on cell apoptosis and TNF receptor super family 6 (Fas)/caspase-8 pathway in rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS).
METHODS:
FLS isolated from the synovial tissue of RA patients were cultured and identified using immunofluorescence staining. The cells were treated with 10% blank serum (blank control group), 10% sera containing low, moderate or high doses of DGNTD, or 20 μmol/mL KR-33493 (a Fas inhibitor) combined with 10% serum containing high-dose DGNTD. MTT assay was used to detect the proliferation of the cells after the treatments. Apoptosis of the cells was detected at 48 h in each group using Hoechst 33342 staining and flow cytometry with annexin V-FITC/PI staining. The mRNA and protein expressions of Fas, FADD, caspase-8 and caspase-3 in the cells at 48 h were detected using qPCR and Western blotting.
RESULTS:
Immunofluorescence staining identified the cultured cells as FLS. Treatment with DGNTD-containing sera significantly inhibited the proliferation of FLS, and the inhibitory effects were enhanced as the dose and intervention time increased ( < 0.05). Hoechst 33342 staining and flow cytometry showed that the sera containing different doses of DGNTD significantly promoted apoptosis of FLS ( < 0.05). The expression levels of Fas, FADD, caspase-8, and caspase-3 at both mRNA and protein levels were significantly increased in the cells after treatment with different doses of DGNTD-containing sera ( < 0.05). The application of KR-33493 obviously reversed the effects of DGNTD on the FLS ( < 0.05).
CONCLUSIONS
DGNTD can induce apoptosis of the FLS by activating Fas/caspase-8 signaling pathway.
Apoptosis
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Arthritis, Rheumatoid
;
Caspase 8
;
Cell Proliferation
;
Cells, Cultured
;
Fibroblasts
;
Synovial Membrane
;
Synoviocytes
5.Treatment of Respiratory Diseases with Banxia Houputang: A Review
Jiren AN ; Xinyue YANG ; Jixian SONG ; Qi CHEN ; Cuiling JIA ; Mengfan SUN ; Yashuo ZHAO ; Ensheng JI
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(5):236-245
Respiratory diseases are common, frequently-occurring clinical diseases. As the prevalence rate is increasing year by year, they have become a problem that seriously affects public health. The diseases are mainly located in the lung by traditional Chinese medicine (TCM) syndrome differentiation. Lung governs Qi and controls breathing and is also an organ for the storage of phlegm. Clinically, phlegm and Qi are often used for the treatment. Banxia Houputang (BHT), originated from Synopsis of the Golden Chamber (《金匮要略》), was used to treat plum-stone Ai (globus hystericus) at first. It is composed of Rhizoma Pinelliae, Cortex Magnoliae Offcinalis, Poria, Rhizoma Zingiberis Recens, and Folium Perillae, and treats diseases with the core pathogensis of mutual obstruction of phlegm and Qi. BHT has the effects of moving Qi, dissipating mass, descending adverse Qi, and resolving phlegm, which basically correspond to the pathological characteristics of the lungs. Clinical studies have confirmed that modified BHT can be used either alone or in combination with western medicine to treat chronic pharyngitis, asthma, chronic obstructive pulmonary disease, pneumonia, obstructive sleep apnea, upper airway cough syndrome and other respiratory diseases, with significant effects. It effectively improves the symptoms and signs of the diseases and reduces the recurrence rate. Basic research has shown that BHT plays anti-inflammatory, anti-oxidative stress, anti-apoptotic, autophagy-regulating, and iron overload-regulating roles by regulating the targets in multiple pathways. This paper, by combing the relevant literature in recent years, conducted a systematic review on BHT from the three aspects of syndrome analysis, clinical treatment research and mechanism research, with a view to providing theoretical basis and reference for the mechanism research of BHT in treating respiratory diseases and for expanding its clinical application.