Circular RNA CircSLC8A1 contributes to osteogenic differentiation in hBMSCs via CircSLC8A1/miR-144-3p/RUNX1 in periprosthetic osteolysis

Circular RNAs (circRNAs) are often found in eukaryocyte and have a role in the pathogenesis of a variety of human disorders. Our related research has shown the differential expression of circRNAs in periprosthetic osteolysis (PPOL). However, the involvement of circRNAs in the exact process is yet unknown. CircSLC8A1 expression was evaluated in clinical samples and human bone marrow mesenchymal stem cells (hBMSCs) in this investigation using quantitative real-time PCR. In vitro and in vivo studies were conducted to explicate its functional role and pathway. We demonstrated CircSLC8A1 is involved in PPOL using gain- and loss-of-function methods. The association of CircSLC8A1 and miR-144-3p, along with miR-144-3p and RUNX1, was predicted using bioinformatics. RNA pull-down and luciferase assays confirmed it. The impact of CircSLC8A1 in the PPOL-mouse model was also investigated using adeno-associated virus. CircSLC8A1 was found to be downregulated in PPOL patients' periprosthetic tissues. Overexpression of CircSLC8A1 promoted osteogenic differentiation (OD) and inhibited apoptosis of hBMSCs in vitro. The osteogenic markers of RUNX1, osteopontin (OPN) and osteocalcin (OCN) were significantly upregulated in hBMSCs after miR-144-3p inhibitor was transferred. Mechanistic analysis demonstrated that CircSLC8A1 directly bound to miR-144-3p and participated in PPOL through the miR-144-3p/RUNX1 pathway in hBMSCs. Micro-CT and quantitative analysis showed that CircSLC8A1 markedly inhibited PPOL, and osteogenic markers (RUNX1, OPN and OCN) were significantly increased (P<0.05) in the mice model. Our findings prove that Circ SLC8A1 exerted a regulatory role in promoting osteogenic differentiation in hBMSCs, and CircSLC8A1/miR-144-3p/RUNX1 pathway may provide a potential target for prevention of PPOL.     

Crystal structure of a staphylokinase: variant a model for reduced antigenicity.

Staphylokinase (SAK) is a 15.5-kDa protein from Staphylococcus aureus that activates plasminogen by forming a 1 : 1 complex with plasmin. Recombinant SAK has been shown in clinical trials to induce fibrin-specific clot lysis in patients with acute myocardial infarction. However, SAK elicits high titers of neutralizing antibodies. Biochemical and protein engineering studies have demonstrated the feasibility of generating SAK variants with reduced antigenicity yet intact thrombolytic potency. Here, we present X-ray crystallographic evidence that the SAK(S41G) mutant may assume a dimeric structure. This dimer model, at 2.3-A resolution, could explain a major antigenic epitope (residues A72-F76 and residues K135-K136) located in the vicinity of the dimer interface as identified by phage-display. These results suggest that SAK antigenicity may be reduced by eliminating dimer formation. We propose several potential mutation sites at the dimer interface that may further reduce the antigenicity of SAK.     

Gene expression,X-inactivation,and methylation during spermatogenesis: The case of Zfa,Zfx, and Zfy in mice

While it has become clear that X-inactivation in the female soma is complete in mouse (in contrast to being “patchy” in man), the degree of X-inactivation in the testes has not been ascertained. We have compared autosomal and X-linked zinc finger homolog expression and X-linked and Y-linked zinc finger homolog methylation in an attempt to elucidate this question. Using RTPCR, we have extended earlier studies of Zfx and Zfa expression in developing testes and find that Zfa expression starts at the time of X-inactivation while Zfx expression is continuous. Cell separation studies did not preclude continued expression of Zfx in adult germ cells. The methylation status of four CCGG residues in the Zfx promoter was studied using PCR bridging this region before and after DNA digestion with the isoschizomers Msp I and Hpa II, the latter being methylation sensitive. Hpa II resistant Zfx promoter DNA was found in all female tissues, but not in male tissues, including the testes. Previous studies have shown that Zfy is expressed at meiosis (like Zfa and unlike Zfx). Despite its expression, the Zfy gene is adjacent to, or contains, highly methylated CCGG sites since hybridization after Msp I digestion detected multiple small fragments that were not released after DNA digestion with Hpa II. Thus, Zfx is not methylated in sperm, while Zfy is, in contrast to their apparent patterns of expression. © 1993 Wiley-Liss, Inc.     

Glycogen synthase kinase 3β inhibition synergizes with PARP inhibitors through the induction of homologous recombination deficiency in colorectal cancer

Monotherapy with poly ADP-ribose polymerase (PARP) inhibitors results in a limited objective response rate (≤60% in most cases) in patients with homologous recombination repair (HRR)-deficient cancer, which suggests a high rate of resistance in this subset of patients to PARP inhibitors (PARPi). To overcome resistance to PARPi and to broaden their clinical use, we performed high-throughput screening of 99 anticancer drugs in combination with PARPi to identify potential therapeutic combinations. Here, we found that GSK3 inhibitors (GSK3i) exhibited a strong synergistic effect with PARPi in a panel of colorectal cancer (CRC) cell lines with diverse genetic backgrounds. The combination of GSK3β and PARP inhibition causes replication stress and DNA double-strand breaks, resulting in increased anaphase bridges and abnormal spindles. Mechanistically, inhibition or genetic depletion of GSK3β was found to impair the HRR of DNA and reduce the mRNA and protein level of BRCA1. Finally, we demonstrated that inhibition or depletion of GSK3β could enhance the in vivo sensitivity to simmiparib without toxicity. Our results provide a mechanistic understanding of the combination of PARP and GSK3 inhibition, and support the clinical development of this combination therapy for CRC patients.Subject terms: Cancer therapeutic resistance, Targeted therapies     

国家公园遴选标准的国际经验及对我国的启示

建立以国家公园为主体的自然保护地体系，是我国生态文明思想的重大举措。国家公园作为国际普遍认可的自然保护手段，受到世界各国的欢迎。美国、加拿大、德国、南非、日本等国的国家公园建设较早、发展较好，通过分析其国家公园建设经验得出：（1）国家公园是以自然保护为首要目标，并为公众提供享受大自然的场所；（2）其评价对象与标准主要包括，自然景观/自然遗迹、生态系统、生物多样性/重要栖息地、面积/范围、自然区域/自然环境、文化景观等；（3）根据资源条件和目标定位的差异，国外国家公园的选建模式主要有基于生态区划的国家公园建设模式、基于社区共建的国家公园建设模式和基于资源主导的国家公园建设模式。在国际经验研究和分析的基础上，结合我国自然资源状况及国家公园建设需求，提出我国国家公园发展建议：（1）明确国家公园的功能定位和发展目标；（2）制定国家公园评价准则和方法，建立以国家代表性、完整性、原真性为主，包括生态区位重要性、历史文化价值等因素的国家公园评价指标体系；（3）划定面向国家公园规划布局的生态地理分区，在区域内建立国家公园，并与生态系统优先区、生物多样性保护关键区，或代表性自然景观格局相协调。     

Nanoantenna Integrated Thermomechanical Infrared Detector

Plasmonics - We numerically studied the nanoantenna integrated thermomechanical mid-infrared detector working at room temperature based on bi-material structures. Three separate bilayer...     

Effects of simulated warming on soil ammonia-oxidizing bacteria and archaea communities in an alpine forest of western Sichuan,China

Ongoing climate change, characterized by winter warming, snow cover decline and extreme weather events, is changing terrestrial ecosystem processes in high altitude and latitude regions. Winter soil processes could be particularly sensitive to climate change. In fact, winter warming and snow cover decline are interdependent in cold biomes, and have a synergistic effect on soil processes. Soil microorganisms not only play crucial roles in material cycling and energy flow, but also act as sensitive bio-indicators of climate change. However, little information is available on the effect of winter warming on forest soil ammonia-oxidizing bacteria (AOB) and archaea (AOA). The alpine and subalpine forest ecosystems on the eastern Tibet Plateau have important roles in conserving soil, holding water, and maintaining biodiversity. To understand the changes in AOB and AOA communities under climate change scenarios, an altitudinal gradient experiment in combination with soil column transplanting was conducted at the Long-term Research Station of Alpine Forest Ecosystems, which is situated in the Bipeng Valley of Lixian County, Sichuan, China. Thirty intact soil columns under an alpine forest at an altitude of 3582 m were transplanted and incubated at 3298 m and 3023 m forest sites, respectively. Compared with the 3582 m, we expected air temperature increases of 2 °C and 4 °C at the 3298 m and 3023 m, respectively. However, the temperatures in the soil organic layer (OL) and mineral soil layer (ML) increased by 0.27 °C and 0.13 °C, respectively, at 3023 m and ? 0.36 °C and ? 0.35 °C at 3298 m. Based on a previous study and with simultaneous monitoring of soil temperature, the abundances of AOB and AOA communities in both the OL and ML were measured by qPCR in December 2010 (i.e., the onset of the frozen soil period) and March 2011 (i.e., the late frozen soil period). The soil columns incubated at 3023 m had relatively higher AOB abundances and lower AOA/AOB ratios than those at 3298 m, while higher AOA abundances and AOA/AOB ratios were observed at 3298 m. The abundance of the microbial community at the late frozen period was higher than that at the onset of frozen soil, and the changes in microbial community abundance at the late frozen period were more substantial. Furthermore, the nitrate nitrogen (N) concentrations in both the OL and ML were significantly higher than ammonia N concentrations, implying that soil nitrate N is the primary component of the inorganic N pool in the alpine forest ecosystem. Additionally, the responses of AOA and AOB in the soil OL to soil column transplanting were more sensitive than the responses of those in ML. In conclusion, climate warming alters the abundance of the ammonia-oxidizing microbial community in the alpine forest ecosystem, which, in turn, might affect N cycling.