Differentiation of isolated human umbilical cord mesenchymal stem cells into neural stem cells

AIM：To investigate whether umbilical cord human mesenchymal stem cell（UC-MSC）was able to differentiate into neural stem cell and neuron.·METHODS：The umbilical cords were o btained from pregnant women with their written consent and the approval of the Clinic Ethnics Committee.UC-MSC were isolated by adherent culture in the medium contains 20%fetal bovine serum（FBS）,then they were maintained in the medium contain 10%FBS and induced to neural cells in neural differentiation medium.We investigated whether UC-MSC was able to differentiate into neural stem cell and neuron by using flow cytometry,reverse transcriptase-polymerase chain reaction（RT-PCR）and immunofluorescence（IF）analyzes.·R ESULTS：A substantial number of UC-MSC was harvested using the tissue explants adherent method at about 2wk.Flow cytometric study revealed that these cells expressed common markers of MSCs,such as CD105（SH2）,CD73（SH3）and CD90.After induction of differentiation of neural stem cells,the cells began to form clusters;RT-PCR and IF showed that the neuron specific enolase（NSE）and neurogenic differentiation 1-positive cells reached 87.3%±14.7%and 72.6%±11.8%,respectively.Cells showed neuronal cell differentiation after induced,including neuron-like protrusions,plump cell body,obviously and stronger refraction.RT-PCR and IF analysis showed that microtubule-associated protein 2（MAP2）and nuclear factor-M-positive cells reached 43.1%±10.3%and 69.4%±19.5%,respectively.·CONCLUSION：Human umbilical cord derived MSCs can be cultured and proliferated and differentiate into neural stem cells,which may be a valuable source for cell therapy of neurodegenerative eye diseases.

Effects of Chinese herbal medicine on plasma glucose,protein and energy metabolism in sheep

Background： The use of antibiotics in animal diets is facing negative feedback due to the hidden danger of drug residues to human health. Traditional Chinese herbal medicine has been used to replace antibiotics in the past two decades and played an increasingly important role in livestock production. The present study was carried out to assess the feeding effects of a traditional nourishing Chinese herbal medicine mixture on kinetics of plasma glucose, protein and energy metabolism in sheep. Ruminal fermentation characteristics were also determined. Methods： Four sheep were fed on either mixed hay （MH-diet） or MH-diet supplemented with 2% of Chinese herbal medicine （mixture of Astragalus root, Angelica root and Atractylodes rhizome; CHM-diet） over two 3S-day periods using a crossover design. The turnover rate of plasma glucose was measured with an isotope dilution method using [U-^13C]glucose. The rates of plasma leucine turnover and leucine oxidation, whole body protein synthesis （WBPS） and metabolic heat production were measured using the [1-^13C]leucine dilution and open circuit calorimetry. Results： Body weight gain of sheep was higher （P = 0.03） for CHM-diet than for MH-diet. Rumen pH was lower （P = 0.02）, concentration of rumen total volatile fatty acid tended to be higher （P = 0.05） and acetate was higher （P = 0.04） for CHM-diet than for MH-diet. Turnover rates of plasma glucose and leucine did not differ between diets. Oxidation rate of leucine tended to be higher （P = 0.06） for CHM-diet than for MH-diet, but the WBPS did not differ between diets. Metabolic heat production tended to be greater （P = 0.05） for CHM-diet than for MH-diet. Conclusions： The sheep fed on CHM-diet had a higher body weight gain and showed positive impacts on rumen fermentation and energy metabolism without resulting in any adverse response. Therefore, these results suggested that the Chinese herbal medicine mixture should be considered as a potential feed additive for sheep.

Efficacy of internal limiting membrane peeling for diabetic macular edema after preoperative anti-vascular endothelial growth factor injection

AIM:To explore the efficacy of minimally invasive vitrectomy(MIV)with or without internal limiting membrane(ILM)peeling on the treatment of diabetic macular edema(DME)in proliferative diabetic retinopathy(PDR)combining with preoperative anti-vascular endothelial growth factor(anti-VEGF)injection.METHODS:Totally 132 eyes(132 patients)diagnosed PDR with DME were included between June 2015 and June 2018 in Tianjin Eye Hospital.The single MIV treatment group included 68 eyes and the MIV combined with ILM peeling group included 64 eyes.Anti-VEGF drugs were injected intravitreally 1wk before the operation and the period of follow-up was 1 to 3y.Best-corrected visual acuity(BCVA),central retinal thickness(CRT),total macular volume(TMV),macular edema(ME)severity,intraocular pressure(IOP),and complications were recorded.Prognostic factors of visual acuity following ILM peeling were analyzed.RESULTS:The BCVA was higher than preoperative values at 1,3,6,and 12mo after surgery in both groups(all P<0.05).At 6 and 12mo,the BCVA of the combined group was significantly higher than that of the MIV only group(0.52±0.23 v/s 0.64±0.29 IogMAR,P=0.011 in 6mo;0.41±0.25\/s 0.52±0.25 IogMAR,P=0.008 in 12mo).Mean CRT values postoperative were significantly lower than preoperative values in both groups from the 1^(st) month(lmo 397.65±106.18 vs 451.94±118.88 μm in MIV only group;388.88±108.68 v/s 464.36±111.53 μm in combined group;both P<0.05)and decreased gradually.The differences between the two groups were statistically significant at 3,6,and 12mo(P=0.004,0.003,0.00 respectively).The TMV was decreased from the 3^(rd) month in the single treatment group(3mo 11.14±1.66 vs 12.20±2.09 mm^(3),P<0.05).At 12mo,the proportion of eyes with edema that had CRT more than 350μm was significantly lower than before surgery(13.24%vs 77.94%in MIV only group;1.56%vs 81.25%in combined group;both P<0.05).There was no significant difference in the recurrence incidence of macular epiretinal membrane,ME,transient IOP increase,vitreous rebleeding,or traction retinal detachment between the two groups.BCVA after ILM excision was positively correlated with the CRT and ME degree before and after surgery(r=0.430,0.485,respectively;P<0.05).CONCLUSION:MIV combined with ILM peeling accelerates the absorption of ME,improves vision,reduces the postoperative CRT and TMV,and reduces the recurrence rate of postoperative ME.