Objective:To explore the regulatory effects of proliferation and apoptosis on THC-8307 by MMS2 siRNA and P53 siRNA.Methods:We experimentally suppressed the MMS2 and P53 expression in human colon cancer cells by the interference RNA technology ( RNAi) as monitored by Real-time qRT-PCR and Western blot.THC-8307 cells that express rate significantly reduced were collected as case group , while using untreated cells as the blank control group , and mock-treated cells as the negative control group.After separately interfering the target genes in each group ,test the relationship and expression level of the two genes.Utilizing flow cytometry techniques to test cells proliferation and apoptosis rate of each group.Results: Compared to the control group , colon cancer cells in which MMS2 and P53 were silenced displayed significant increase of P53,MMS2 mRNA and protein levels(P<0.05).MMS2-depleted cells displayed increase in apoptosis rates ,for both early and later stages ( P<0.05 ).Conclusion: MMS2 and P53 negatively regulate each other in colon cancer cells proliferation and apoptosis .

OBJECTIVE：To investigate the correlation between the morphological characteristics and chemical constituents of garden ginseng （GG），mountain transplanted ginseng （MTG）and mountain-grown ginseng （MGG），so as to provide evidence for elucidating the scientific connotation of “evaluating quality from morphological characteristics ”. METHODS ：A total of 30 batches of GG ，MTG and MGG were collected. The contents of 11 saponins（ginsenoside Rg 1，Re，Rg2，F1，Rb1，Rb2，Rb3，Rc，Rd，Ro， Rf）were determined by UPLC. The determination was performed on Acquity UPLC ® BEH C 18 column with mobile phase consisted of 0.1％ Formic acid solution- 0.1％ Acetonitrile formate solution （gradient elution ）at the flow rate of 0.5 mL/min. The column temperature was set at 35 ℃. The detection wavelength was set at 203 nm，and sample size was 5 μL. The content of starch was determined by colorimetry ，and the absorbance value was measured at 559 nm with 0.5％ iodine-potassium iodide as the color reagent. SPSS 23.0 statistical software was used to perform Pearson relationship analysis for appearance characteristic indexes （number of stem scars ，rhizome length ，texture）with the contents of 11 kinds of saponins ，the content of total saponins ，the content ratio of panaxatriol saponin （PPT）/panaxadiol saponin （PPD），the content of starch. RESULTS ：The linear relationship between UPLC and colorimetry was good （r＞0.999）. RSDs of precision ，reproducibility and stability tests were all lower than 5％ . Average recoveries were 95.17％ -105.20％（RSD＜5％ ，n＝6）. Except for ginsenoside Rf ，the contents of other 10 ginsenosides were positively correlated with the number of stem scars ，so were the contents of 11 gensenosides with the rhizome length（r＞0，P＜0.05），and they were negatively correlated with the firmness of the texture （r＜0，P＜0.05）. The content of starch in P. ginseng was positively correlated with texture firmness （r＝0.95，P＜0.01）. The ratio of PPT/PPD in P. ginseng was negatively associated with the number of stem scars and the rhizome length （r＜0，P＜0.05）. CONCLUSIONS ：The number of stem scars ，the rhizome length and texture porosity of P. ginseng are positively associated with the contents of medicinal material ginsenoside；the increase of PPD content in P. ginseng was higher than that of PPT with the increase of growth years. The traditional“evaluating quality from morphological characteristics ”for P. ginseng has a certain scientific connotation ，which is more stem scars ，longer rhizome and loose texture.

Severe muscle injury is hard to heal and always results in a poor prognosis. Recent studies found that extracellular vesicle-based therapy has promising prospects for regeneration medicine, however, whether extracellular vesicles have therapeutic effects on severe muscle injury is still unknown. Herein, we extracted apoptotic extracellular vesicles derived from mesenchymal stem cells (MSCs-ApoEVs) to treat cardiotoxin induced tibialis anterior (TA) injury and found that MSCs-ApoEVs promoted muscles regeneration and increased the proportion of multinucleated cells. Besides that, we also found that apoptosis was synchronized during myoblasts fusion and MSCs-ApoEVs promoted the apoptosis ratio as well as the fusion index of myoblasts. Furthermore, we revealed that MSCs-ApoEVs increased the relative level of creatine during myoblasts fusion, which was released via activated Pannexin 1 channel. Moreover, we also found that activated Pannexin 1 channel was highly expressed on the membrane of myoblasts-derived ApoEVs (Myo-ApoEVs) instead of apoptotic myoblasts, and creatine was the pivotal metabolite involved in myoblasts fusion. Collectively, our findings firstly revealed that MSCs-ApoEVs can promote muscle regeneration and elucidated that the new function of ApoEVs as passing inter-cell messages through releasing metabolites from activated Pannexin 1 channel, which will provide new evidence for extracellular vesicles-based therapy as well as improving the understanding of new functions of extracellular vesicles.
Creatine/metabolism* ; Extracellular Vesicles ; Muscle, Skeletal/metabolism* ; Myoblasts/metabolism* ; Regeneration ; Connexins/metabolism*