The effect of reduced glutathione on the chondrogenesis of human umbilical cord mesenchymal stem cells

It has been discussed whether reduced glutathione (GSH) could promote the chondrogenic differentiation ability of human umbilical cord mesenchymal stem cells (hUC-MSCs). hUC-MSCs were isolated from human umbilical cord and their specificity was identified, then induced into cartilage-like cells in chondrogenic induction medium with transforming growth factor beta 1 (TGF-β1), especially with GSH. The morphological change before and after induction was observed through inverted phase contrast microscope, Type II collagen (COL2-A1) and glycosaminoglycan (GAG) were analyzed qualitatively by Toluidine blue and immunofluorescence technique, respectively, the contents of COL2-A1 and GAG were estimated from the determination of hydroxyproline content and Alcian Blue method separately. The mRNA expressions of GAG and COL2-A1 were assayed by real-time fluorescence quantitative PCR. After continuously cultured for 21 days with GSH, Toluidine blue staining and immunofluorescence reaction were all positive in basic induction medium group (group B), basic induction medium +0.5% dimethylsulfoxide (DMSO) group (group BD) and basic induction medium +0.5% DMSO +500 μM GSH group (group BDG). Moreover, compared with group B and group BD, the contents of COL2-A1 and GAG in group BDG relatively increased and the mRNA expression level of COL2-A1 and GAG also comparatively increased (P < 0.05) and both had a significant statistical significance (P < 0.05). So GSH might promote the induction of hUC-MSCs to differentiate into cartilage-like cells.

Transcriptome analysis of blood stasis syndrome in subjects with hypertension

OBJECTIVE:To screen for m RNAs associated with blood stasis syndrome and to explore the genetic mechanisms of blood stasis syndrome in hypertension.METHODS:This study involved groups of patients with hypertension and blood stasis,including those with Qi deficiency,Qi stagnation,cold retention and heat retention;as well as hypertensive patients without blood stasis and healthy individuals.Human umbilical vein endothelial cells were co-cultured with the sera of these healthy individuals and patients with blood stasis syndrome.Total RNA was extracted from these cells and assessed by a high-throughput sequencing method(Solexa)and digital gene expression.Differentially expressed genes among these six groups were compared using whole genome sequences,and m RNAs associated with blood stasis syndrome identified.Differences in gene use and gene ontology function were an-alyzed.Genes enriched significantly and their pathways were determined,as were network interactions,and encoded proteins.Gene identities were confirmed by real-time polymerase chain reactions.RESULTS:Compared with cells cultured in sera of the blood stasis groups,those culture in sera of healthy individuals and of the non-blood stasis group showed 11 and 301 differences,respectively in stasis-related genes.Genes identified as differing between the blood stasis and healthy groups included activating transcription factor 4,activating transcription factor 3,DNA-damage inducible transcription factor 3,Tribbles homolog 3,CCAAT/enhancer binding protein-β,and Jun proto-oncogene(JUN).Pathway and protein interaction network analyses showed that these genes were associated with endoplasmic reticulum stress.Cells cultured in sera of patients with blood stasis and Qi deficiency,Qi stagnation,heat retention,and cold retention were compared with cells cultured in sera of patients with the other types blood stasis syndrome.The comparison showed differences in expression of 28,28,34,and 32 specific genes,respectively.CONCLUSION:The pathogenesis of blood stasis syndrome in hypertension is related to endoplasmic reticulum stress and involves the differential expression of the activating transcription factor 4,activating transcription factor 3,DNA-damage inducible transcription factor 3,Tribbles homolog 3,CCAAT/enhancer binding protein-β,and JUN genes.

Differential Expression of MicroRNA in Endothelial Cells Incubated with Serum of Hypertension Patients with Blood-Stasis Syndrome

Objective： To screen out blood-stasis syndrome （BSS）-associated microRNA and therefore determine the possible target for treating hypertension. Methods： A high-energy sequencing method and digital gene expression sequencing theory were adopted to sequence microRNA （miRNA） and messenger RNA （mRNA）, and to determine differential expression in human umbilical vein endothelial cells incubated with serum samples from hypertension patients with or without BSS, and healthy controls. The results were confirmed using gene prediction software. Results： A total of 13 miRNAs and 11 mRNAs showed statistical difference both in the BSS/normal groups and BSS/non-BSS groups, respectively. Four pairs of target mRNNmiRNA were identified： FRMD4Nhsa-miR-34a, MAP3K14/hsa-miR-34a, PER1/hsa-miR-34a, and FGF2/hsa-miR-132. Conclusion： Four mRNNmiRNA pairs mentioned above seem to be involved in pathogenesis and maintenance of hypertension with BSS.

Effect of Kangshuai Yizhi Formula Ⅰ (抗衰益智方Ⅰ) on Learning and Memory Dysfunction Induced by Scopolamine in Mice

Objective：To evaluate the improvement of Kangshuai Yizhi FormulaⅠ（抗衰益智方Ⅰ,KYFⅠ）on the learning and memory dysfunction in mice,and on the mechanism of the hippocampal cholinergic system and the nervous system of monoamine which are closely related to learning and memory function.Methods：Mice in the low-,middle-,and high-dose KYFⅠgroups were given low-,middle-,and high-dose KYF,respectively, by gastrogavage for 35 successive days.Animals in the control group and the model group were treated with distilled water.The acute learning and memory dysfunction model was established by injection of scopolamine from day 31,and Morris water maze was used to assess the behavior performance of scopolamine-induced model mice for five days.The activities of acetylcholinesterase（AChE）,choline acetyl transferase（ChaT） and the content of monoamine neurotransmitters in hippocampus were measured.The activity of monoamine oxidase（MAO）in hippocampus and serum was also detected.Results：（1）Compared with the control group,the mean escape latency was shortened,and the frequency across the platform and the staying time at the platform area on the 5th day were decreased in the model group by Morris water maze test.The activities of AChE and MAO were increased,and the ChaT activity and monoamine neurotransmitter content were decreased as well.（2） The escape latency for 4 days in the low-,middle-,and high-dose KYFⅠgroups was significantly shortened than that in the model group,with the shortest latency in the high-dose KYFⅠgroup（P〈0.05,P〈0.01）.The frequency across the platform was significantly increased and the staying time at the platform was significantly prolonged in the middle-and high-dose KYFⅠgroups（P〈0.05,P〈0.01）.（3）As compared with the model group,the activity of ChaT and the content of monoamine neurotransmitters in the hippocampus were significantly increased, and the activities of AchE and MAO were significantly decreased in the hippocampus in the high-dose KYFⅠgroup（P〈0.01）.Conclusions：High-dose KYFⅠcan significantly improve the learning and memory dysfunction induced by scopolamine in mice.Its mechanism may be related to improving the central cholinergic system and regulating the hippocampal monoamine neurotransmitters.

Planar cell polarity genes, Celsr1-3, in neural development

flamingo is among the ＇core＇ planar cell-polarity genes, protein of which belongs to a unique cadherin subfamily. In contrast to the classic cadherins, composed of several extracellular cadherin repeats, one transmembrane domain and one cytoplasmic segment linked to catenin binding, Drosophila Flamingo has seven transmembrane segments and a cytoplasmic tail with no catenin-binding sequence. In Drosophila, Flamingo has pleotropic roles in controlling epithelial polarity and neuronal morphogenesis. Three mammalian orthologs of flamingo, Celsrl-3, are widely expressed in the nervous system. Recent work has shown that Celsrl-3 play important roles in neural development, such as in axon guidance, neuronal migration, and cilium polarity. CeIsrl-3 single-gene knockout mice exhibit different phenotypes, but there are cooperative interactions among these genes.