Bone marrow mesenchymal stem cells (BMSCs) are a group of pluripotential non-hematopoietic somatic stem cells niched in bone marrow.With the characteristics of stable genetic traits,pluripotential in differentiation,easy to isolate from source tissue,and fast to proliferate when cultured in vitro,BMSCs are currently attracting extensive research interests,and considered to be one of the most promising candidates in corneal tissue engineering.At present,many research groups,domestic and abroad,have reported that BMSCs can not only differentiate into corneal limbal stem cells,corneal epithelial cells,and corneal endothelial cells,but also play an important role in ocular surface repair.However,the successful application of BMSCs in cornea usually depends on the correct selection of supporting materials or scaffold,such as xenogeneic corneal stroma and amniotic membrane.Other unsolved problems in BMSCs-related corneal tissue engineering include the molecular biologic mechanism underlying the directional differentiation from BMSCs to corneal cells,the standards to identify BMSCs from differentiated corneal cells,the optimal scaffold materials and the potential tumorigenicity with grafting of transformed or undifferentiated BMSCs.This paper reviewed the progresses and issues of corneal tissue engineering with BMSCs.

Objective To investigate the clinical effect of orthokeratology (ortho-k)during night and wearing frameglass during daytime for treating high myopic in teenagers.Methods 53 teenager cases (aged 10 - 14 years old)of high myopia with spherical equivalent degree -6.00--8.00 diopters (D)in our hospital were performed the retrospective analysis,among them,23 cases were the combined wearing of ortho-k and frameglass and 30 cases were simple wearing of frameglass.The changes of eye axi-al length at 6,12,18 24 months before and after wearing frameglass were measured and compared between the two groups.The spherical equivalent degrees were compared between before wearing frameglass and at 2 years after wearing frameglass.Results The spherical equivalent degree after stopping wearing for 1 month after 2-year wearing in the ortho-k and frameglass group was(-7.28±0.57)D,which after 2 years in the frameglass group was (-7.80±0.68)D,the 2-year spherical equivalent degrees in the or-tho-k and frameglass group and the frameglass group were increased by (-0.29 ±0.22)D and(-1.07 ±0.41)D respectively,the difference between the two groups was statistically significant(P <0.01);the eye axial growths were(0.12±0.04)mm and(0.37± 0.13)mms respectively,the difference between the two groups was statistically significant (P <0.01).Conclusion The combina-tion of ortho-k during night and frameglass during daytime can effectively relieve the myopia increase and eye axis elongation in the patients with high myopia and control the myopia development to some extent.

Objective: To investigate the nootropic components of Hovenia dulcis. Methods: Six saponins were isolated from Hovenia dulcis and Step-down test was used to examine the memory ability of mice. The escape latency and the times of wrong performance within 3 min were used to evaluate the memory ability of mice. To study the effects of saponins on learning and memory in mice, we divided the mice into 6 groups: youth group, aged group, aged plus piracetam(0.3 g/kg) group, aged plus saponins (0.6 g/kg) group, aged plus saponins (0.3 g/kg) group, and aged plus saponins(0.15 g/kg) group. To study the influence of saponins on impairment of memory acquirement, consolidation, and reoccurrence (induced by scopolamine, sodium nitrite and 40% ethanol, respectively), mice were also divided into the following 7 groups: control group, untreated group, Piracetam group (0.3 g/kg), compound 3 group(0.3 g/kg), compound 4 group(0.3 g/kg), compound 5 group(0.3 g/kg), and compound 6 group (0.3 g/kg). Results: The chemical structures of six saponins were elucidated as 3-O-stigmasterol-(6-O- palmitoyl)-β-D-glucopyranoside (1), β-daucosterin (2), hovenidulcioside A1 (3), hoduloside I (4), hoduloside IV (5), and saponins C2 (6). Among them, compounds 1 and 2 were isolated from this plant for the first time. Compounds 5 and 6 had enhancing effect on the learning and memory ability of natural senile mice, and they could improve the impairment of memory acquirement, consolidation and recurrence in mice induced by scopolamine, sodium nitrite and 40% ethanol, respectively. Conclusion: The aglycone of jujubogenin might be the main saponins contributing to the nootropic effect of total saponins from Hovenia dulcis.

Humans ; Male ; Prostate-Specific Antigen ; blood ; Prostatectomy ; Prostatic Neoplasms ; blood ; metabolism ; radiotherapy ; surgery

Chromatography ; methods ; DNA Fingerprinting ; Drugs, Chinese Herbal ; analysis ; Magnetic Resonance Spectroscopy ; Mass Spectrometry

Objective To investigate the effects of epigenetic drugs on 2D- and 3D-cultured cholangiocarcinoma cells in vitro.Methods In this study,we have built compact and round TFK-1 spheroid in poly-HEMA coated 96-well plate and determined the effects of epigenetical drugs on 2D and 3D cultured cholangiocarcinoma cells:TFK-1.Viability of 2D and 3D cells model was examined by WST assay and FDA/PI staining. Using methylation-specific PCR analysis,we demonstrated the changes of methylation status of promoters regarding three tumor suppressor genes APC,E-Cadherin,and p16 INK4A.Results The average diameters of compact and round TFK-1 spheroids were in the range of 350-400 μm.The TFK-1 spheroid cells were more resistant to the epigenetic drugs and demonstrated a 11.2155-fold higher IC50 values for hydralazine and valproic acid than the same cells grown as monolayer. Higher doses of epigenetic drugs were needed to reverse the hypermethylation status in 3D cultured cells than 2D cells; however,the parallel dosage - dependent characteristic did not show in the 3D spheroid group.Conclusions Taken together,we established a 3D culture model of human cholangiocarcinoma epithelial spheroid.The 3D spheroid cells are more complex than the 2D monolayer cells and their unique characteristics are able to affect the consequences of epigenetic therapy.The 3D spheroid is a promising model for the research of epigenetic therapy.

Objective To explore the effects of octylphenol, an environmental contaminant with estrogenic activity, on the expression of apoptosis regulator gene bcl-2 mRNA in human breast cancer cell line MCF-7 and to compare its effects with 17?-estradiol (E 2). Methods MCF-7 cells were cultured in vitro and exposed to various concentrations of octylphenol and E 2 from 12 h to 120 h respectively. Total RNA was abstracted and the expression of bcl-2 mRNA was measured using reverse transcription polymerase chain reaction (RT-PCR) approach. Results The expression of bcl-2 mRNA in MCF-7 cells increased significantly after exposure to octylphenol and E 2 24 h and the effects continued to 120 h. The stimulation effect on the expression of bcl-2 mRNA was induced by octylphenol with a wide range of concentrations, and the strongest effect was found at the concentration of 10-6 mol/L of octylphenol. Conclusion The results suggested that octylphenol had similar estrogenic activities as E 2 on regulating the expression of bcl-2 mRNA in MCF-7 cells, but the effect was weaker than that of E 2.

Background Inflammation and adipogenesis are two parallel processes with increasing activity in severe thyroid-associated ophthalmopathy(TAO),and mevastatin was proved to have the inhibiting effect on the differentiation of adipose.Objective The aim of this work was to investigate the effects of mevastatin on the expression of cyclooxygenase-2(COX-2)and peroxisome proliferator activated receptor-γ(PPAR-γ)and differentiation of TAO-derived orbital preadipocytes,and explore its modulation effects on lipopolysaccharide(LPS)induced inflammation and the differentiation of TAO-derived orbital preadipocytes in vitro.Methods The retroorbital adipose tissue was obtained from 4 TAO patients during the surgery.The orbital fibroblasts were cultured from orbital adipose tissues using explant culture method.To study the suppressing effect of mevastatin on inflammatory response,cultured cells were divided into 5 groups.The 1000 μg/L LPS orbital fibroblasts were stimulated for 8 hours in group A,and 1000 μg/L LPS combined with 5 μmol/L,10 μmoL/L or 20 μmoL/L mevastatin were used respectively for the substitute in the group B,group C and group D.The orbital fibroblasts in group E were cultured routinely without any intervention as control.To observe the inhibiting effect of mevastatin on the differentiation of adipose,the group A were then subdivided into group A1-A6.After 1000 μg/L LPS was used to treat the cells for 8 hours,the ceils were induced to differentiate into adipocytes.All orbital preadipocytes from A1 to A6 were stimulated to differentiate into mature adipocytes with cocktail differentiation medium for a 10-day duration.During the procedure,group A2,A3 and A4 were interfered with 5,10 or 20 μmol/L mevastatin,and in the group A5 and A6,10 μmol/L mevastatin were added at the fourth day or eighth day.Intracellular fat accumulation in differentiated adipocytes was determined by oil red O staining.The absorption(A492 nm)was measured in the ceils by enzyme-linked immunosorbent assay(ELISA).Expression of COX-2 and PPAR-γ mRNA was detected by reverse transcription polymerase chain reaction(RT-PCR),and the expression of COX-2 and PPAR-γ protein was detected by Westernblot.The level of PGE2 in the supernatant was detected by ELISA.Results The expression of COX-2 protein and mRNA as well as the PGE2 levels in B,C,D group decreased markedly in comparison with those in A group(P＜0.05).With the increase of mevastatin concentration,the expression of COX-2 protein and mRNA as well as the PGE2 levels in B,C,D groups decreased successively(F =228.380,101.745,1586.881,P＜0.05).The expression of COX-2 protein and mRNA and PGE2 levels in E group were lower significantly than those in A,B and C groups(P＜0.05),but no significant differences were found between E group and D group(P＞0.05).The A492 value and the expressions of PPAR-γ protein and mRNA in differentiated cells showed the successively decrease in A1-A4 group with the elevation of mevastatin concentration(P＜0.05),and the evidently decreased A492 value and the expressions of PPAR-γ protein and mRNA also were seen in A1 and A5 groups compared with A3 group(P ＜ 0.05).Conclusions Mevastatin inhibits LPS-induced COX-2 expression,PPAR-γ expression,PGE2 secretion and differentiation of TAO-derived orbital fibroblasts in vitro in dose-dependent manner.Mevastatin plays these effect more prominently in early stage of adipocytes differentiation.

Objective To study the related influencing factors of MCF 7 cells proliferation test in vitro to provide some criteria for the standard detection of environmental estrogens (EE) by cell proliferation test of MCF 7 cells in vitro . Methods The sensitivity to E 2 stimulation of different sources of MCF 7 cell lines was detected. Cell proliferation test was performed on the screened sensitive cells in different culture conditions. Results In the estrogen free medium, C cell line of MCF 7 was more sensitive to E 2 stimulation than A and B cell lines ( P

Animals ; Betaine-Homocysteine S-Methyltransferase ; therapeutic use ; Homocysteine ; blood ; Hyperhomocysteinemia ; blood ; therapy ; Male ; Rats ; Rats, Wistar ; Recombinant Proteins ; therapeutic use