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The in vitro culture of Blastocystis hominis in Roswell Park Memorial Institute (RPMI) 1,640 medium containing 20% calf serum is described. The morphological and reproductive mode studies of B. hominis in symptomatic patients' faeces and in further RPMI 1,640 medium-cultured samples were undertaken by light microscopy using iodine staining and hematoxylin staining. Three distinct morphological forms, vacuolar, granular and amoeboid, were distinguished in faeces and in vitro cultures. The cystic form was detected in long-term cultures. Five modes of reproduction, namely, binary fission, endodyogeny, plasmotomy, budding and schizogeny were observed.

Background The development of Systemic lupus erythematosus (SLE) has been associated with the balance of Th17 and Treg cells. IL-2 and rapamycin can influence the populations of both Th17 and Treg cells. However, it is unclear whether low dose of IL-2 and rapamycin can relieve the symptoms of SLE patients and what is the mechanisms. In this study, we aim to analyze the effect of low dose of IL-2 plus rapamycin on the number of Tregs, Th17 cells and the ratio of Th17/Treg cells, as well as to evaluate its therapeutic efficacy in refractory SLE patients. Result Fifty refractory SLE patients and 70 healthy controls were enrolled and followed up for 24 weeks. We found that compared with HC, the refractory SLE patients had a lower number of Tregs, a similar number of Th17 cells, but an increased ratio of Th17/Treg. After the treatment, the number of Tregs of the patients at 12th and 24th week was significantly increased. While the number of Th17 cells was unchanged, the ratio of Th17/Treg was significantly decreased at both 6 weeks and 24 weeks. After 6, 12 and 24 weeks of treatment, the SLEDAI score was significantly reduced. The prednison dosage at 6th,12th and 24th week post treatment was significantly decreased. Conclusion Our results support that the reduction of Tregs and the imbalance of Th17/Treg cells were correlated with the occurrence and development of refractory SLE. Low dose of IL-2 combined with rapamycin was able to restore the number of Tregs and the balance of Th17/Treg cells. As a result, this approach was able to induce immune tolerance and promote disease remission, allowing for the reduction in prednisone dosage. Trial registration ChiCTR-IPR-16009451 Registration date: 2016/10/16