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Objective@#To investigate the effects of artesunate treatment on chronic graft-versus-host disease (cGVHD).@*Methods@#Recipient BALB/c mice received 8 × 106 bone marrow cells with 8×106 spleen cells from B10D2 mice. Artesunate solubilized in acetone was injected intraperitoneally every day at the dose of 1 mg/kg at Day 28 after BMT. The clinical scores, survival and histopathological damage were analyzed. The frequency of Th17 and Tregs in PB and spleens from the mice were evaluated by flow cytometry. In addition, CD4+ T cells from the spleens of mice were cultured in vitro, then stimulated with artesunate, the frequency of Th17 and Tregs in these splenocytes were evaluated by flow cytometry.@*Results@#Artesunate administration diminished clinical and histopathological damage, and improved the survival of cGVHD mice[(46.57±7.83)% vs (55.71±6.99)%, χ2=5.457, P=0.020]; Artesunate contributed to Tregs development [(4.45±0.04)% vs (8.40±0.23)%, t=15.679, P<0.001; (6.62±0.24)% vs (10.48±0.48)%, t=6.587, P=0.003] while decreased Th17 cells [(1.51±0.18)% vs (0.58±0.19)%, t=7.233, P<0.001; (1.48±0.38)% vs (0.71±0.18)%, t=3.653, P=0.011] expressions in both PB and spleens, and decreased the Th17/Treg ratio (0.34±0.05 vs 0.09±0.03, t=7.621, P=0.002; 0.19±0.03 vs 0.06±0.02, t=6.993, P=0.002). Moreover, artesunate suppressed the Th17 cells expressions [(0.82±0.37) % vs (3.39±1.22) %, t=4.044, P=0.007] and contributed to Tregs development [(34.63±1.29) % vs (14.28±1.69) %, t=19.119, P<0.001], and also decreased the Th17/Treg ratio (0.24±0.09 vs 0.02±0.01, t=4.780, P=0.003) in vitro.@*Conclusions@#Artesunate suppressed the Th17 cells expressions and contributed to Tregs development, which provided new sights into the development of a novel drug for cGVHD, e.g., artemisinin.
ABSTRACT
AIM:To investigate the expression and potential role of complement 5a receptor (C5aR) in chro-nic graft-versus-host disease (cGVHD).METHODS:The expression of C5aR on lymphocytes and the frequency of CD4+CD25+ Foxp3+ regulatory T cells (Tregs) in 20 cGVHD patients and 9 healthy donors was detected by flow cytometry.The correlation between the expression of C5aR and the percentage of Tregs in the cGVHD patients was analyzed.In addition, the splenocytes from the mice were cultured in vitro, and stimulated these splenocytes with recombinant mouse C5a protein (rmC5a).The peripheral blood mononuclear cells (PBMCs) from cGVHD patients were cultured in vitro, which was inhibited by C5aR antagonist (C5aRA).The frequency of Tregs in these splenocytes and the PBMCs were evaluated by flow cytometry.RESULTS:The expression of C5aR on the lymphocytes was significantly increased in the cGVHD patients compared with the healthy donors, while the percentage of Tregs was markedly lower in the cGVHD patients.The expression of C5aR was negatively correlated with the percentage of Tregs.Furthermore, the development of Tregs was suppressed by rmC5a stimulation, but was promoted by C5aRA in vitro.CONCLUSION:C5aR elevation is associated with Treg reduction in cGVHD, indicating that C5aR may play a potential role in suppressing Tregs, resulting in the incidence of cGVHD.
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A 13-year-old girl suddenly developed hypertrophic rough-surfaced plaques on both elbows, knees and ankles without obvious precipitating factors at 3 years of age.Because there was no subjective symptom or other discomfort, no treatment was given.Thereafter, the lesions neither faded nor spread.Skin examination revealed lesions covered with 1-2 mm-sized keratotic papules on the elbows and knees, which were clustered together and confluent in some areas.Circular keratotic plaques were observed on the ankles.Histopathology showed epidermal hyperkeratosis with mild parakeratosis, follicular keratotic plugs, acanthosis, broadened dermal papillae, telangiectasis and perivascular mononuclear infiltration in dermal papillae, infiltration of a few nonspecific cells in the superficial dermis.According to the clinical manifestations and histopathological findings, the patient was diagnosed with keratosis circumscripta, and treated with oral vitamin A and topical halometasone cream and urea-vitanmin E cream.The degree of skin hypertrophy and roughness was decreased after 15 days of treatment, but increased 1 month after drug withdrawal.
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Objective:To study an identifying model of HPLC fingerprint for the overall quality control of traditional Chinese medicine. Methods: A HPLC method is used to establish the standard fingerprint as well as sample fingerprint first. Then through the principles of numerical taxonomy and statistics, the similarity of the sample to the standard is tested by coefficients of cosine and distant, which are calculated with computer. Results: The combination of the two coefficients of cosine and distant can test the similarity of the sample to the standard correctly and accurately. Conclusion: The coefficients along with HPLC fingerprint can be applied to the on-line control in workshop to ensure the products' quality of all batches.