Differentiation potential of rabbit CD90-positive cells sorted from adipose-derived stem cells in vitro

To investigate the differentiation potential of purified CD90⁺ cells sorted from adipose-derived stem cells (ADSCs), CD90⁺ cells were sorted from rabbit ADSCs using flow cytometry. Then, cell expansion of CD90⁺ cells and unsorted ADSCs was observed using an inverted microscope. Furthermore, cell surface markers including CD40, CD105, and CD90 on CD90⁺ cells and unsorted ADSCs were quantified using flow cytometry. Additionally, multi-lineage differentiation ability between CD90⁺ cells and unsorted ADSCs was compared, and expression of adipocyte-related genes PPAR-r and CEBPA as well as stem cell-related gene SOX2 in CD90⁺ cells and unsorted ADSCs was determined using real-time quantitative PCR. We found that CD90⁺ cells had a stronger cell proliferation ability than unsorted ADSCs. CD90⁺ cells showed a stronger ability of osteoblast and chondrocyte differentiation than unsorted ADSCs and CD90^? cells, whereas the adipose differentiation ability of CD90⁺ cells was similar to that of ADSCs and CD90^? cells. CD14, CD105, and CD90 on CD90⁺ cells were expressed more highly than those on ADSCs. Additionally, the mRNA expression level of SOX2 in CD90⁺ cells was significantly higher than that in ADSCs, whereas the expression of PPAR-r and CEBPA was markedly lower than that in ADSCs. These results suggested that the purified CD90⁺ cells sorted from ADSCs exhibit a stronger differentiation potential than the unsorted ADSCs.     

Crystal structure of Usutu virus envelope protein in the pre-fusion state

Background

Usutu virus (USUV) is a mosquito-born flavivirus that can infect multiple avian and mammalian species. The viral surface envelope (E) protein functions to initiate the viral infection by recognizing cellular receptors and mediating the subsequent membrane fusion, and is therefore a key virulence factor involved in the pathogenesis of USUV. The structural features of USUV-E, however, remains un-investigated thus far.

Findings

Using the crystallographic method, we determined the structure of USUV-E in the pre-fusion state at 2.0?angstrom. As expected, the overall fold of USUV-E, with three β-barrel domains (DI, DII, and DIII), resembles those of other flaviviral E proteins. In comparison to other pre-fusion E structures, however, USUV-E exhibits an apparently enlarged inter-domain angle between DI and DII, leading to a more extended conformation. Using our structure and other reported pre-fusion E structures, the DI-DII domain-angle difference was analyzed in a pairwise manner. The result shows a much higher degree of variations for USUV-E, indicating the potential for remarkable DI-DII domain angle plasticity among flaviviruses.

Conclusion

We report the crystal structure of USUV-E and show that its pre-fusion structure has an enlarged DI-DII domain-angle which has not been observed in other reported flaviviral E-structures.

     

内蒙古高原不同生境条件下甘蒙锦鸡儿水分调节特性和抗逆性的比较研究

对分布于内蒙古和林格尔（半干旱地区）和阿拉善（强干旱地区）地区的甘蒙锦鸡儿种群水分调节特性和抗逆性进行了比较研究.结果表明,阿拉善种群比和林格尔种群的叶片渗透调节物质含量高、渗透势低、渗透调节能力强,叶片含水量和自由水含量低、束缚水含量和束缚水/自由水比值高,叶水势和气孔导度低,表明阿拉善种群比和林格尔种群有更强的水分调节能力.阿拉善种群丙二醛(MDA)含量大于和林格尔种群,但细胞膜相对透性和超氧自由基含量小于和林格尔种群,表明阿拉善种群自由基清除能力强、细胞膜稳定性高.有效的水分调节能力和较强的抗逆性是甘蒙锦鸡儿适应干旱环境的重要生理基础.     

Protective Role of Deoxyschizandrin and Schisantherin A against Myocardial Ischemia–Reperfusion Injury in Rats

Background

Our previous studies suggested that deoxyschizandrin (DSD) and schisantherin A (STA) may have cardioprotective effects, but information in this regard is lacking. Therefore, we explored the protective role of DSD and STA in myocardial ischemia–reperfusion (I/R) injury.

Methodology/Principal Findings

Anesthetized male rats were treated once with DSD and STA (each 40 µmol/kg) through the tail vein after 45 min of ischemia, followed by 2-h reperfusion. Cardiac function, infarct size, biochemical markers, histopathology and apoptosis were measured and mRNA expression of gp91^phox in myocardial tissue assessed by RT-PCR. Neonatal rat cardiomyocytes were pretreated with DSD and STA and then damaged by H₂O₂. Cell apoptosis was tested by a flow cytometric assay. Compared with the I/R group: (i) DSD and STA could significantly reduce the abnormalities of LVSP, LVEDP, ±dp/dt_max and arrhythmias, thereby showing their protective roles in cardiac function; (ii) DSD and STA could significantly attenuate the infarct size and MDA release while increasing SOD activity, suggesting a role in reducing myocardial injury; (iii) tissue morphology and myocardial textual analysis revealed that DSD and STA mitigated changes in myocardial histopathology; (iv) DSD and STA decreased apoptosis (33.56±2.58% to 10.28±2.80% and 10.98±1.99%, respectively) and caspase-3 activity in the myocardium (0.62±0.02 OD/mg to 0.38±0.02 OD/mg and 0.32±0.02 OD/mg, respectively), showing their protective effects upon cardiomyocytes; and (v) DSD and STA had similar protective effects on I/R injury as those seen with the positive control metoprolol. In vitro, DSD and STA could significantly decrease the apoptosis of neonatal cardiomyocytes.

Conclusions/Significance

These data suggest that DSD and STA can protect against myocardial I/R injury. The underlining mechanism may be related to their role in inhibiting cardiomyocyte apoptosis.     

The complete mitochondrial genome of Neobenedenia melleni (Platyhelminthes: Monogenea): mitochondrial gene content,arrangement and composition compared with two Benedenia species

The complete mitochondrial (mt) genome sequences of Neobenedenia melleni were determined and compared with those of Benedenia seriolae and B. hoshinai. This circular genome comprises 13,270 bp and includes all 36 typical mt genes found in flatworms. Total AT content of N. melleni is 75.9 %. ATG is the most common start codon, while nad4L is initiated by GTG. All protein-coding genes are predicted to terminate with TAG and TAA. N. melleni has the trnR with a TCG anticodon, which is the same to B. seriolae but different from B. hoshinai (ACG). The mt gene arrangement of N. melleni is similar to that of B. seriolae and B. hoshinai with the exception of three translocations (trnF, trnT and trnG). The overlapped region between nad4L and nad4 was found in the N. melleni mt genome, which was also reported for the published Gyrodactylus species, but it was not found in those of B. seriolae and B. hoshinai, which are non-coding regions instead. The present study provides useful molecular characters for species or strain identification and systematic studies of this parasite.     

Overexpression of monocarboxylate anion transporter 1 and 4 in T24-induced cancer-associated fibroblasts regulates the progression of bladder cancer cells in a 3D microfluidic device

Stromal fibroblasts are essential for tumor proliferation and invasion. Here we presented a 3-dimensional (3D) microfluidic co-culture device to reconstruct an in vivo-like tumor microenvironment for investigation of the interactions of cancer-associated fibroblasts (CAFs) and bladder cancer cells. With this device, we verified that the cytokines secreted by bladder cancer cells T24 effectively transform the fibroblasts into CAFs. Compared to fibroblasts, the CAFs, which undergo the aerobic glycolysis, showed higher ability to produce lactate and provide energy for bladder cancer cell proliferation and invasion. We also demonstrated that this kind of tumor-promoting effect was associated with the upregulation of monocarboxylate anion transporter 1 (MCT1) and MCT4 expression in CAFs. We concluded that MCT1 and MCT4 are involved in bladder cancer cell proliferation and invasiveness. Moreover, this 3D microfluidic co-culture device allows for the assay to characterize various cellular events in a single device sequentially, facilitating a better understanding of the interactions among heterotypic cells in a sophisticated microenvironment.     

Atomic‐Scale Spacing between Copper Facets for the Electrochemical Reduction of Carbon Dioxide

Copper (Cu) offers a means for producing value‐added fuels through the electrochemical reduction of carbon dioxide (CO₂), i.e., the CO₂ reduction reaction (CO₂RR), but designing Cu catalysts with significant Faradaic efficiency to C₂₊ products remains as a great challenge. This work demonstrates that the high activity and selectivity of Cu to C₂₊ products can be achieved by atomic‐scale spacings between two facets of Cu particles. These spacings are created by lithiating CuO_x particles, removing lithium oxides formed, and electrochemically reducing CuO_x to metallic Cu. Also, the range of spacing (d_s) is confirmed via the 3D tomographs using the Cs‐corrected scanning transmission electron microscopy (3D tomo‐STEM), and the operando X‐ray absorption spectra show that oxidized Cu reduces to the metallic state during the CO₂RR. Moreover, control of d_s to 5–6 Å allows a current density exceeding that of unmodified CuO_x nanoparticles by about 12 folds and a Faradaic efficiency of ≈80% to C₂₊. Density functional theory calculations support that d_s of 5–6 Å maximizes the binding energies of CO₂ reduction intermediates and promotes C–C coupling reactions. Consequently, this study suggests that control of d_s can be used to realize the high activity and C₂₊ product selectivity for the CO₂RR.     

血管内皮细胞生长因子复性条件研究

利用大肠杆菌 BL2 1 (DE3)表达血管内皮细胞生长因子 (VEGF16 5)表达蛋白以包含体形式存在。为了获得有生物活性的蛋白质 ,我们对影响复性的参数 :氧化型、还原型谷胱甘肽比例 ,复性液的 p H值 ,复性时间 ,精氨酸浓度 ,血管内皮细胞生长因子起始浓度进行了较为系统的研究 ,初步获得了具有一定生物活性的 VEGF16 5二聚体蛋白。