1.Iinhibitory effects of miR-200a on proliferation and migrating ability of conjunctival fibroblasts and its mechanism
Xue, YIN ; Ya, LIANG ; Zhilan, YUAN
Chinese Journal of Experimental Ophthalmology 2016;34(12):1087-1091
Background Scarring of surgical area,the most important factor,leads to the failure of glaucoma filtering surgeries.Therefore,more and more attentions are paid to the causes and process of scar formation.Objective This study was to compare the differences of proliferation and migrating abilities of fibroblasts between filtering bleb scar tissue and normal Tenon capsular tissue,and to investigate the inhibitory effects of miRNA-200a (miR-200a) on biological behavior of conjunctival fibroblasts.Methods Normal Tenon capsular tissue and filtering bleb scar tissue were collected during the strabismus surgery and glaucoma filtering surgery,respectively for the primarily culture of fibroblasts.The proliferation (absorbency,A) of the cells was assayed by cell counting kit-8 (CCK8) method;the relative migrating distance of the cells was measured by cell scratch test;and the relative expressions of transforming growth factor-β1 (TGF-β1)mRNA and miR-200a mRNA in the cells were detected by realtime fluorescence quantitative PCR.TGF-β1 mimic of 0,1,2 and 5 ng/ml was added in the medium of human normal Tenon capsular-derived fibroblasts (HTFs),and 0.00,0.25,0.50,1.00 μg/ml TGF-β1 inhibitor was added in the medium of human scarring-derived fibroblasts (HSFs) for 24 hours,respectively,and CCK8 was used to evaluate the proliferation of the cells.The relative migrating distance as well as the relative expressions of miR-200a mRNA were analyzed in the 2 ng/ml TGF-β1 mimic-or 1.00 μg/ml TGF-β1 inhibitor-treated cells.Results The primary conjunctival presented the spindle and star-like in shape with large body and oval nuclei.The cells showed the positive response for keratin and vimentin antibodies.The A values were 1.476±0.110 in the HSFs and 0.958±0.074 in the HTFs,with a significant difference between them (t =24.900,P=0.016).The relative expressions of TGF-β1 mRNA were significantly higher in the HSFs than those in the HTFs,and the relative expressions of miR-200a were evidently lower in the HSFs than those in the HTFs,showing significant differences between them (t =6.358,P =0.024;t=7.394,P =0.018).Compared with the 2 ng/ml TGF-β1 mimic-treated HTFs,the relative migrating distance increased,while the expression level of miR-200a mRNA was significantly reduced in the 2 ng/ml TGF-β1 mimictreated HSFs (all at P<0.05);Compared with the 1.00 μg/ml TGF-β1 inhibitor-treated HTFs,the relative migrating distance decreased,but the expression level of miR-200a mRNA was significantly elevated in the 1.00 μg/ml TGF-β1 inhibitor-treated HSFs (all at P<0.05).Conclusions The proliferation and migrating abilities are stronger in the HSFs than those in the HTFs,which probably is regulated by the expression of miR-200a in the cells.The miR-200a plays a negative feedback for the effect of TGF-β1 promoting proliferation and migration of fibroblasts.
2.IN VIVO ~1H NMR SPECTROSCOPY IN RAT C6 GLIOMA
Hong YIN ; Zhilan HUANG ; Yuangu GAO
Medical Journal of Chinese People's Liberation Army 1983;0(02):-
By using the high magnetic field spectrometer to assess the practical value of 1 H NMR spectroscopy in vivo through observation of changes in metabolism of C6 glioma, fourty Wister rats were divided into two groups: ten as controls, and in thirty rats C6 glioma cells were implanted spheroids in right caudate nucleus. MR images were carried out at intervals. The 1 H NMR spectroscopy were performed by using point resolved spectroscopy (PRESS) technique. The NAA Cho, Cr+PCr, Glu+Gln, Lip, Lac were observed in normal rats. The NAA intensity was seen to be decreased or disappeared in C6 glioma, and NAA/Cho ratio and NAA/Cr ratio decreased obviously( P
3.Survival of bone marrow mesenchymal stem cells after transplantation into the rat infarcted myocardium
Chuwei LIN ; Shenghua ZHOU ; Haiying DAI ; Ping DENG ; Hongguang HUANG ; Zhilan YIN ; Xiansong GUAN
Chinese Journal of Tissue Engineering Research 2014;(41):6628-6632
BACKGROUND:The preliminary findings confirm that bone marrow mesenchymal stem celltransplantation is safe and effective in the treatment of acute myocardial infarction, but its exact mechanism is unclear. There are few studies addressing the survival status of transplanted stem cells and its acting timing.
OBJECTIVE:To study the survival of rat bone marrow mesenchymal stem cells transplanted into the infracted myocardium. METHODS:Bone marrow mesenchymal stem cells were cultured using density gradient centrifugation. Eighty rat models of myocardial infarction were prepared. Bone marrow mesenchymal stem cells were injected via a microsyringe at four sites around the infarcted region at 14 days after modeling. Then, 70 rats with living cells were selected for detecting the survival of bone marrow mesenchymal stem cells at days 3, 5, 7, 10, 20, 28 after transplantation. RESULTS AND CONCLUSION:Under ×400 visual field, the number of Brdu-positive bone marrow mesenchymal stem cells was (36±12) at 3 days posttranplantation, (33±13) at 5 days, (28±9) at 7 days, (15±5) at 10 days, (5±3) at 14 days, 0 at 20 days, and 0 at 28 days, showing a overal downward trend after transplantation. The number of bone marrow mesenchymal stem cells was negatively correlated with transplant days (P<0.01, r=-0.47). The number of cells decreased most significantly within 1 week after transplantation, and then decreased to 0 at 20 days. These findings indicate that transplanted bone marrow mesenchymal stem cells in the myocardium cannot survive for a long term and also cannot be transformed into myocardial tissue.
4.Efficacy and metabolic safety of long-term treatment with ethinyl oestradiol/cyproterone and desogestrel/ethinyl oestradiol tablets in women with polycystic ovary syndrome.
Jun ZHANG ; Mi SU ; Liangzhi XU ; Zhilan YANG ; Weiyao YIN ; Ying NIE ; Xiaoyong QIAO ; Ran CHENG ; Yaxian MA
Journal of Southern Medical University 2018;38(8):917-922
OBJECTIVETo evaluate the efficacy and metabolic safety of long-term treatment with ethinyl oestradiol/cyproteroneand desogestrel/ethinyl oestradiol tablets in women with polycystic ovary syndrome (PCOS).
METHODSWomen with PCOSfrom West China Second Hospital of Sichuan University enrolled between September, 2011 and August, 2013 were randomlyallocated to receive either ethinyl oestradiol/cyproterone tablets (Group A, =355) or desogestrel/ethinyl oestradiol tablets(Group B, =357) for a prospective observation period of 6 months. Women with insulin resistance also received metformin. Atbaseline, 3 months, and 6 months, the patients were evaluated for menstruation, acne score, body mass index (BMI), waist-tohip ratio (WHR), plasma levels of sex hormones, fasting blood glucose (FPG), HOMA-insulin resistance index (HOMA-IR), serum lipid, ovarian volume, and the number of ovarian follicles.
RESULTSAll the patients had a regular menstrual cycle aftertreatments. Testosterone level, acne score, LH/FSH, ovarian volume, and the number of follicles decreased significantly afterthe treatments without significant differences between the two groups. Significant increases were noted in TG, TCh, LDL, HDL, and AIP, and HDL level in group A as compared with group B ( < 0.001). FPG decreased in both groups, and wassignificantly lower in group B at 6 months ( < 0.05). BMI and WHR decreased in all the patients with insulin resistance aftercombination treatment with metformin ( < 0.05), but increased significantly in patients without insulin resistance ( < 0.05). Ingroup A, HOMA- IR significantly increased in patientswithout insulin resistance at 3 months ( < 0.05), whereas asignificant increase was not observed until 6 months ingroup B ( < 0.05).
CONCLUSIONSBoth ethinyl oestradiol/cyproterone tablets and desogestrel/ethinyl oestradioltablets can relieve the symptoms of PCOS, but it isadvisable to assess the risk of cardiovascular diseasebefore the treatments.