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OBJECTIVE@#To explore the effect of miR-335-5p/ADCY3 interaction on the lymphocyte function in the patients with aplastic anemia (AA).@*METHODS@#Blood samples were collected from 22 healthy volunteers (HC) and 50 AA patients including 38 severe AA (SAA) and 12 non-severe AA (NSAA). Peripheral blood mononuclear cells (PBMNC) were isolated. The expression of miR-335-5p and ADCY3 mRNA was detected by using RT-PCR. Negative control miR-335-5p (NC group) and miR-335-5p mimic (mimic group) were transfected to AA-PBMNC by using RNAimax reagent, respectively. The proliferative ability, activation and cytokines of CD4 T and CD8 T cells were measured by flow cytometry. Dual-luciferase reporter assay was used to verify the targeted relationship between miR-335-5p and target gene.@*RESULTS@#The expression of miR-335-5p was significantly downregulated in SAA-PBMNC and NSAA-PBMNC compared with HC-PBMNC (0.08±0.01 vs 0.74±0.10, P＜0.01; 0.17±0.02 vs 0.74±0.10, P＜0.01). Meanwhile, the expression of miR-335-5p in SAA-PBMNC was very statistically significantly lower than that in NSAA-PBMNC (P＜0.01). Compared with NC group, upregulation of miR-335-5p in vitro could significantly inhibited the proliferation of CD4 T and CD8 T cells in AA-PBMNC (P＜0.05 and P＜0.05, respectively). And, upregulating miR-335-5p in AA-PBMNC could significantly inhibited the activation of CD4 and CD8 T cells (P＜0.01 and P＜0.01, respectively). The ratio of CD4TNFα T, CD8IFNγ+T and CD8TNFα T cell by up-regulating the expression of miR-335-5p from AA-PBMNC in vitro was also significantly lower (P＜0.01, P＜0.05 and P＜0.05, respectively). In addition, the expression of ADCY3 was higher in AA-PBMNC than that in HC-PBMNC (1.70±0.15 vs 0.76±0.12, P＜0.01). Furthermore, by means of dual-luciferase reporter assay, the luciferase activity of ADCY3'UTR wildtype could be inhibited by miR-335-5p.@*CONCLUSIONS@#The expression of miR-335-5p was significantly downregulated in AA, and that correlates with disease severity. Up-regulating miR-335-5p can correct the hyperimmune status in AA patients by targeting ADCY3. These changes may relates with the strengthen of inhibition for targeted gene ADCY3.
Subject(s)
Humans , Anemia, Aplastic , Genetics , CD8-Positive T-Lymphocytes , Leukocytes, Mononuclear , Lymphocyte Count , MicroRNAs , GeneticsABSTRACT
Fifteen compounds were isolated from the toad skin by a combination of various chromatographic methods including macroporous resin, silica gel, ODS and semi-preparative HPLC. Their structures were identified as 4,5-dimethyl-1,3,4,5-tetrahydropyrrolo[4,3,2-de]quinolin-6-ol(1), serotonin(2), N-methyl serotonin(3), O-methyl bufotenine(4), 1,2,3,4-tetrahydro-6-hydroxy-β-carboline(5), O-methylserotonin(6), glycinebetaine(7), caffeine(8), bufotenine(9), shepherdine(10), tryptophan(11), (5-hydroxy-1H-indol-3-yl)acetic acid(12), 5-hydroxy tryptophol(13), 2-methyl-6-hydroxy-1,2,3,4-tetrahydro-β-carboline(14), bufothionine(15). Among them, compound 1 was a new compound,compound 5 was a new natural product. Compounds 4-8 and 10-14 were separated from toad skin for the first time.
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AIM:To investigate the effect of suramin concentration changes on trabeculectomy in rabbit, and to provide treatment strategies for glaucoma on the basis of experiment. METHODS:Thirty-two albino rabbits were randomly divided into four groups, including standard control group, experimental group Ⅰ, experimental group II, and experimental group Ⅲ. Each eye was performed standard trabeculectomy. During surgery, standard control group was given a piece of cotton with 0. 3mg/mL mitomycin C ( MMC ) for 2min, and the other three groups were given a piece of cotton with 0. 3, 0. 4, and 0.5mg/mL suramin respectively for 2min. The filtering blebs and intraocular pressure ( IOP ) were observed at the 3, 7, 15, and 30d after surgery. Some conjunctiral specimen were observed with hitochemicall ( HE staining) and immunohistochemicall methods. RESULTS:At postoperative 7, 15, and 30d, the changes of the IOP, functional filtering blebs, and the number of positive cell nuclear in experimental group II and experimental group Ⅲ were significantly different compared with those in standard control group and experimental group Ⅰ (all P0. 05). The changes of the IOP and the number of positive cell nuclear in experimental group Ⅲ were significantly different compared with those in experimental group II (P0. 05). The status of filtering channel in experimental groupII and experimental group Ⅲ were better than those in experimental group Ⅰ and standard control group. CONCLUSION: The concentration of suramin has a significantly influence on its effect. When the concentration is 0. 3mg/mL, the antiproliferative effect of suramin is no more than that of MMC. The effect of 0. 4, 0.5mg/mL suramin is better than MMC. 0. 5mg/mL suramin has a better effect on controlling IOP and suppressing the growth of fibroblasts than 0. 4mg/mL suramin.
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AIM:To observe the rehabilitation effect of oral Calcium Dobesilate Tablets and Epalrestat Tablets on the proliferative diabetic retinopathy ( PDR ) patients after vitrectomy. METHODS:After vitrectomy patients informed consent, they were randomly divided into intervention group and control group, and their vision and fundus were followed up and compared at 2d;3, 6, 9, 12mo after operation. RESULTS: The recovery and sustainment of visual acuity, and subsiding of macular edema of patients in drug intervention group were significantly better than that in control group. CONCLUSION: PDR patients after vitrectomy actively orally take medicine to protect retina, which can improve vision and fundus rehabilitation.
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<p><b>OBJECTIVE</b>To compare therapeutic effects of acupoint application of Chinese medicine and routine acupuncture on bronchial asthma.</p><p><b>METHODS</b>One hundred cases were randomly divided into an observation group and a control group, 50 cases in each group. The observation group was treated with application of Chinese medicine at Dazhui (GV 14), Feishu (BL 13), Gaohuang (BL 43), Danzhong (CV 17), Qihai (CV 6) in the dog days; and the control group was treated with acupuncture at Zhongfu (LU 1), Tiantu (CV 22), Danzhong (CV 17), Dingchuan (EX-B 1), Fenglong (ST 40). The long-term therapeutic effects were observed.</p><p><b>RESULTS</b>The total effective rate was 94.0% in the observation group and 76.0% in the control group with a significant difference between the two groups (P < 0.05).</p><p><b>CONCLUSION</b>The therapeutic effect of acupoint application of Chinese medicine in the dog days on bronchial asthma is significantly superior to that of routine acupuncture.</p>