Objective:To detect collagen I synthesis activity in the vitreous of PVR induced by macrophages in rabbits.
Methods:PC Ⅲ (Procollagen Ⅲ ) concentrations were measured by radioim-
munoassay in the vitreous samples of 14 rabbit eyes with experimental PVR and 14 control eyes.
Results:The mean PC Ⅲ concentration on the 7th day after macrophage injection as 257.58mu;g/L(range,236.04~266.88mu;g/L,n= 4)and significantly increased on the 14th day later. On the 28th day the mean concentration of PC Ⅲ as 912.23mu;g/L (range, 881.36~943.10mu;g/L ;n= 2). There was a significant difference between the 7th and the 14th, 21st of 28th day statistically(P＜0.05). PC Ⅲ was not detected in control eyes.
Conclusion:The PC Ⅲ level in the vitreous of rabbit eyes with experimental
PVR increased significantly from the 7th to the 28th day after macrophages injection and is well consistent with the time course of scarring and the development of traction retinal detachment in the PVR model.
（Chin J Ocul Fundus Dis,1996,12: 43-44）
ObjectiveTo study the effects of leukemia inhibitory factor (LIF) and basic fibroblast growth factor (bFGF) on the proliferation and differentiation of human bone marrow mesenchymal stem cells (hBMSCs).
MethodshBMSCs at passage 4 were divided into 4 groups according to different culture conditions:cells were treated with complete medium (α-MEM containing 10%FBS, group A), with complete medium containing 10 ng/mL LIF (group B), with complete medium containing 10 ng/mL bFGF (group C), and with complete medium containing 10 ng/mL LIF and 10 ng/mL bFGF (group D). The growth curves of hBMSCs at passage 4 in different groups were assayed by cell counting kit 8; cellular morphologic changes were observed under inverted phase contrast microscope; the surface markers of hBMSCs at passage 8 including CD44, CD90, CD19, and CD34 were detected by flow cytometry.
ResultsThe cell growth curves of each group were similar to the S-shape; the cell proliferation rates in 4 groups were in sequence of group D > group C > group B > group A. Obvious senescence and differentiation were observed very early in group A, cells in group B maintained good cellular morphology at the early stage, with slow proliferation and late senescence; a few cells in group C differentiated into nerve-like cells, with quick proliferation; and the cells in group D grew quickly and maintained cellular morphology of hBMSCs. The expressions of CD44 and CD90 in groups A and C at passage 8 cells were lower than those of groups B and D; the expressions of CD19 and CD34 were negative in 4 groups, exhibiting no obvious difference between groups.
ConclusionLIF combined with bFGF can not only maintain multiple differentiation potential of hBMSCs, but also promote proliferation of hBMSCs.
Objective To observe the effects of keratinocytes on proliferation and collagen secretion of fibroblasts. Methods The conditioned medium,collected from cultured keratinocytes, was added to the cultured fibroblasts as the tested groups(12.5％, 25% and 50％ groups) and DMEM as control group. The MTT, hydroxyproline coloricmetric method and flow cytometer were employed to measure the fibroblast proliferation, the collagen secretion andthe change of the cell cycle.Results In fibroblast proliferation, the absorbency(A) value of tested groups was significantly different from that of the control group (P＜0.01). A value increased as increasing concentration, there was statistically significant difference betweetheconcentrations of 25%,50% and the concentration of 12.5%(P＜0.01), but no statistically significant difference between the concentrations of 25% and 50%(P＞0.01). In collagen secretion, there was no statistically significant difference between the tested groups and the control group(P＞0.01), and between the tested groups(P＞0.01). In cell cycle, 50% of conditioned medium could make the fibroblast pass the limit of G1/S and S/G2 period, the cell rates of S,G2-M period increased. Conclusion The conditioned medium from keratinocytes can increase fibroblasts proliferation, have little effect on general collagen secretion.
Objective To investigate the impacts of neoadjuvant chemotherapy on the expression of insulin-like growth factor-1 receptor (IGF-1R) and on operation procedure and the significance of prognosis. Methods The expression of IGF-1R in 40 patients with breast cancer before and after neoadjuvant chemotherapy was measured by immunohistochemistry. The diagnosis was proved by core biopsy. All the patients took the TAC chemotherapy regimen. Modified radical operation was performed after two chemotherapy cycles and the IGF-1R expression was measured again. The clinical effect of neoadjuvant chemotherapy was assessed according to WHO criterion by measuring the size of tumor by physical examination and B type ultrasound. Results After neoadjuvant chemotherapy the tumor size shrank in 29 patients, there was no CR (complete response) or PD (progressed disease) to be documented. IGF-1R expression could be downregulated in 25 patients. Conclusion Neoadjuvant chemotherapy can inhibit the tumor growth by downregulation of the expression of IGF-1R.
Pulmonary arterial hypertension(PAH) is a kind of pulmonary hypertension disease. Recently, the researches of its pathogenesis have reached more and more deeply. The treatment of pulmonary arterial hypertension is individual and systematic, not only relying on medicine treatment. The treatment of PAH is as follows: common treatment, non-specific medicine treatment, targeted medicine treatment, NO breath-in treatment, gene treatment, intervention and surgery treatment.The article reviews the main treatment of pulmanory arteral hypertesion to provide new thought and evidence in clinic.
Objective To study the effects of adenosine 2A receptor activation on activation, proliferation, and toxicity of T lymphocytes stimulated by phytohemagglutinin (PHA) in vitro. Methods A model of activated T cells was established by stimulating the cells with PHA. Those T cells were treated with different concentrations of adenosine 2A receptors agonist (0.01 μmol/L, 0.1 μmol/L, 1 μmol/L, and 10 μmol/L CGS21680). The expressions of CD69, CD25 and proliferation of T cells were measured by fluorescent antibody stain and flow cytometry. ELISA method was used to detect IL-2 and INF-γ levels. Results All concentrations of CGS21680 significantly inhibited the expressions of CD25 and CD69 on PHA-stimulated T cells surface and proliferation of T cells （Plt;0.05， Plt;0.01）. IL-2 and INF-γ secreted by T cells were significantly suppressed, too （Plt;0.01）. Conclusion Activation of adenosine 2A receptor can effectively inhibit the activation, proliferation, and toxicity of T cells in vitro.
Objective To explore the role of estrogen receptor alpha (ERα) and estrogen receptor beta (ERβ) in estrogen-induced proliferation of endometrial cancer, and explore whether metformin inhibits the proliferation of endometrial cancer cells through ERα and ERβ. Methods Stable transfected Ishikawa cells were constructed by lentivirus. The effects of down-regulated ERα and ERβ on estrogen-induced Ishikawa cell proliferation were detected by methyl thiazolyl tetrazolium assay. The effects of down-regulated ERα and ERβ on estrogen-induced Ishikawa cell cycle were detected by flow cytometry. In addition, quantitative real-time polymerase chain reaction and Western blotting assays were used to detect changes in the expression of cyclinD1 and P21 involved in cell cycle regulation. The effects of down-regulated ERα and ERβ on estrogen-induced Ishikawa cell proliferation were observed by adding metformin to estrogen treatment. Results Down-regulation of ERα inhibited the proliferation and cell cycle of Ishikawa cells (P<0.05). Down-regulation of ERα also inhibited the expression of cyclinD1 and promoted the expression of P21 (P<0.05). Down-regulation of ERα counteracted the effect of estrogen-induced cell proliferation, cell cycle, and the expression changes of cyclinD1 and P21 (P<0.05). Down-regulation of ERβ promoted the proliferation and cell cycle of Ishikawa cells (P<0.05). Down-regulation of ERβ also promoted the expression of cyclinD1 and inhibited the expression of P21 (P<0.05). Down-regulation of ERβ enhanced the effect of estrogen-induced cell proliferation, cell cycle, and the expression changes of cyclinD1 and P21 (P<0.05). Metformin inhibited the proliferation of estrogen-induced Ishikawa cells (P<0.05), while in the down-regulated ERα Ishikawa cells or down-regulated ERβ Ishikawa cells, the inhibition of metformin on Ishikawa cells disappeared (P<0.05). Conclusions ERα may promote estrogen-induced proliferation of endometrial cancer cells, while ERβ may inhibit estrogen-induced proliferation of endometrial cancer cells. In addition, ERα and ERβ may also mediate the inhibitory effect of metformin on endometrial cancer cells.
【Abstract】ObjectiveTo investigate the growth effect of methylprednisolone （MP） on human hepatocellular carcinoma cell line HepG2.Methods Periodic distribution of cells and cellular apoptosis were detected by using cell culture,immunofluorescence staning of Annex Ⅴ and flow cytometric analysis in hepatocellular carcinoma cell.Results Compared with control group, methylprednisolone increased Ｇ0/Ｇ1 phase cell, decreased S phase cell on human hepatocellular carcinoma cell line HepG2 ,which had positive correlation with the time.The apoptosis rate and the necrosis rate of cells had the relation of dose-dependent with the concentration of MP, the cell membrane of early cellular apoptosis was stained green fluorescence. Conclusion Methylprednisolone can induce Ｇ0/Ｇ1 arrest , may play a proliferation-inhibition effect on the hepatocellular carcinoma cell line HepG2.
ObjectiveTo explore the immunosuppressive effect of XGD1 and its mechanism. MethodsDifferent concentrations of XGD1 were added to PHA or ConA induced human peripheral blood T lymphocyte.Seventytwo hours later modified MTT assay was employed to test the effect of XGD1 on T cell proliferation. Flowcytometry was used to examine the effect of XGD1 on the expression of IL2 receptor(IL2R) on the surface of T cells individually at 48 h and 72 h.And the effects of XGD1 combined with cyclosporine A(CsA) on the proliferation of Tlymphocytes and the expression of IL2R were also investigated. ResultsIn the concentration range of 0.2～25 μg/ml,XGD1 exerted marked inhibitory effect on PHA or ConA induced T cell proliferation,which was proportional to dose. Flowcytometry showed that XGD1 inhibited the expression of IL2R,and the percentage of IL2Rα positive cells after stimulation of PHA decreased from 47.67% to 25.03% in the presence of XGD1 (1 μg/ml).XGD1 and CsA had synergism in inhibition of T cell proliferation and IL2R expression. ConclusionThe study suggests that XGD1 has immunosuppressive effect. The suppressive effect of XGD1 on T cell proliferation is most probably mediated by decreasing IL2R expression.
Objective
To investigate the effect of different concentrations of raloxifene (RAL) on the proliferation and apoptosis of human aortic valve interstitial cells (AVICs) in vitro.
Methods
AVICs were isolated from human aortic valve by collagenase type Ⅱ, and cultured in different concentrations (0 nmol/L, 0.1 nmol/L, 1 nmol/L,10 nmol/L, 100 nmol/L and 1 000 nmol/L) of RAL. AVICs cultured in 0 nmol/L RAL were treated as the control group and those in other concentrations of RAL as the experiment groups. The proliferation and apoptosis of AVICs were evaluated by Cell Proliferation Assay (MTS assay) on day 0, 3, 5, 7 and 9. Flow cytometry was used to detect the cell cycle and apoptosis of AVICs on day 7.
Results
MTS results showed that the optical density value at 490 nm was much less in 10 nmol/L RAL and 100 nmol/L RAL groups (P＜0.05) on day 5, 7 and 9 than that in the control group. Flow cytometry results demonstrated that S-phase rate (P＜0.05) and cell apoptosis rate (P＜0.05) on day 7 were lower in the 10 nmol/L and 100 nmol/L RAL groups compared with the control group.
Conclusion
RAL with suitable concentration can inhibit proliferation and apoptosis of AVICs, which will lay an important foundation for further research of the role of RAL on heart valve diseases.
