Using Amino-Labeled Nucleotide Probes for Simultaneous Single Molecule RNA-DNA FISH

Using amino-labeled oligonucleotide probes, we established a simple, robust and low-noise method for simultaneous detection of RNA and DNA by fluorescence in situ hybridization, a highly useful tool to study the large pool of long non-coding RNAs being identified in the current research. With probes either chemically or biologically synthesized, we demonstrate that the method can be applied to study a wide range of RNA and DNA targets at the single-cell and single-molecule level in cellular contexts.     

Altered Resting-State Amygdala Functional Connectivity after 36 Hours of Total Sleep Deprivation

Objectives

Recent neuroimaging studies have identified a potentially critical role of the amygdala in disrupted emotion neurocircuitry in individuals after total sleep deprivation (TSD). However, connectivity between the amygdala and cerebral cortex due to TSD remains to be elucidated. In this study, we used resting-state functional MRI (fMRI) to investigate the functional connectivity changes of the basolateral amygdala (BLA) and centromedial amygdala (CMA) in the brain after 36 h of TSD.

Materials and Methods

Fourteen healthy adult men aged 25.9±2.3 years (range, 18–28 years) were enrolled in a within-subject crossover study. Using the BLA and CMA as separate seed regions, we examined resting-state functional connectivity with fMRI during rested wakefulness (RW) and after 36 h of TSD.

Results

TSD resulted in a significant decrease in the functional connectivity between the BLA and several executive control regions (left dorsolateral prefrontal cortex [DLPFC], right dorsal anterior cingulate cortex [ACC], right inferior frontal gyrus [IFG]). Increased functional connectivity was found between the BLA and areas including the left posterior cingulate cortex/precuneus (PCC/PrCu) and right parahippocampal gyrus. With regard to CMA, increased functional connectivity was observed with the rostral anterior cingulate cortex (rACC) and right precentral gyrus.

Conclusion

These findings demonstrate that disturbance in amygdala related circuits may contribute to TSD psychophysiology and suggest that functional connectivity studies of the amygdala during the resting state may be used to discern aberrant patterns of coupling within these circuits after TSD.     

Cloning and Characterization of tesk1, a Novel Spermatogenesis-Related Gene,in the Tongue Sole (Cynoglossus semilaevis)

Testis-specific protein kinase 1 (Tesk1) is a serine/threonine kinase with unique structural features. In the present study, we cloned and characterized the tesk1 gene of tongue sole, Cynoglossus semilaevis. The full-length tesk1 cDNA consists of 1,672 nucleotides, encoding a 331 amino acid polypeptide with a characteristic structure composed of an N-terminal kinase domain and a C-terminal proline-rich domain. The tesk1 genomic sequence contains eight exons and seven introns. Real-time quantitative PCR revealed that tesk1 mRNA is expressed predominantly in the testis, though the level of expression varied throughout development. We used in situ hybridization to show that tesk1 mRNA is expressed in the spermatids of males and pseudo-males, but not in triploid males. Our results suggest that tongue sole Tesk1 may play a role in spermatogenesis.     

Transplantation of ATP7B–Transduced Bone Marrow Mesenchymal Stem Cells Decreases Copper Overload in Rats

Background

Recent studies have demonstrated that transplantation of ATP7B-transduced hepatocytes ameliorates disease progression in LEC (Long-Evans Cinnamon) rats, a model of Wilson''s disease (WD). However, the inability of transplanted cells to proliferate in a normal liver hampers long-term treatment. In the current study, we investigated whether transplantation of ATP7B-transduced bone marrow mesenchymal stem cells (BM-MSCs) could decrease copper overload in LEC rats.

Materials and Methods

The livers of LEC rats were preconditioned with radiation (RT) and/or ischemia-reperfusion (IRP) before portal vein infusion of ATP7B-transduced MSCs (MSCs^ATP7B). The volumes of MSCs^ATP7B or saline injected as controls were identical. The expression of ATP7B was analyzed by real-time quantitative polymerase chain reaction (RT-PCR) at 4, 12 and 24 weeks post-transplantation. MSC^ATP7B repopulation, liver copper concentrations, serum ceruloplasmin levels, and alanine transaminase (ALT) and aspartate transaminase (AST) levels were also analyzed at each time-point post-transplantation.

Results

IRP-plus-RT preconditioning was the most effective strategy for enhancing the engraftment and repopulation of transplanted MSCs^ATP7B. This strategy resulted in higher ATP7B expression and serum ceruloplasmin, and lower copper concentration in this doubly preconditioned group compared with the saline control group, the IRP group, and the RT group at all three time-points post-transplantation (p<0.05 for all). Moreover, 24 weeks post-transplantation, the levels of ALT and AST in the IRP group, the RT group, and the IRP-plus-RT group were all significantly decreased compared to those of the saline group (p<0.05 compared with the IRP group and RT group, p<0.01 compared with IRP-plus-RT group); ALT and AST levels were significantly lower in the IRP-plus-RT group compared to either the IRP group or the RT group (p<0.01 and p<0.05. respectively).

Conclusions

These results demonstrate that transplantation of MSCs^ATP7B into IRP-plus-RT preconditioned LEC rats decreased copper overload and was associated with an increase in MSC engraftment and repopulation.     

Inferring Novel Indications of Approved Drugs via a Learning Method with Local and Global Consistency

Inferring new indication of approved drugs is critical not only for the elucidation of the interaction mechanisms between these drugs and their associated diseases, but also for the development of drug therapy for various human diseases. This paper proposes a network-based approach to reveal the association between 52 human diseases and potential therapeutic drugs based on multiple types of data. The advantage of the approach is that it can obtain the global relevance features for each drug-disease pair in the network by the learning local and global consistency method (LLGC). Cross-validation tests results demonstrate the proposed approach can achieve better performance comparing with previous methods. More importantly, it provides a promising strategy to maximize the value of therapeutic drugs and offer safe and effective treatments for different diseases.     

甘蔗类异黄酮还原酶(IRL)基因的克隆与表达分析

采用RT-PCR技术从甘蔗中克隆So IRL基因,用生物信息学方法对获得的氨基酸序列进行分析,利用荧光定量PCR技术研究So IRL基因在甘蔗不同组织和不同胁迫条件下的表达特性。结果表明,克隆获得甘蔗So IRL,Gen Bank登录号为KF808324。该c DNA全长1 169 bp,含有1个927 bp的完整开放阅读框(ORF),编码309个氨基酸。系统进化树分析显示,甘蔗So IRL与玉米的IRL蛋白亲缘关系较近。q RT-PCR分析表明So IRL在甘蔗根、茎、叶中均有表达;在RSD病菌及低温(4℃)、聚乙二醇(PEG)、Na Cl和脱落酸(ABA)4种非生物胁迫下均被诱导表达,但表达模式不同。说明该基因可能参与甘蔗应答RSD过程,并可能在非生物胁迫中也发挥了作用。     

大型医院医疗设备采购流程精细化管理的探讨

医疗设备采购工作由于环节众多,过程复杂,且工作性质的特殊性、敏感性关系,很多的审批、监督环节无法省略,对工作速度造成了严重影响。该文探讨了一种基于项目团队模式的医疗设备采购管理新模式。打破了采购工作各相关部门的职能壁垒,按精细化管理的原则,构建了一个跨部门的医疗设备采购工作职能团队,快速、准确、高效地完成大型医院医疗设备购置工作,提高医疗设备采购工作的效率和质量,以充分发掘采购工作的效益。     

Endometrial progesterone resistance and PCOS

Polycystic ovary syndrome (PCOS) is a state of altered steroid hormone production and activity. Chronic estrogen exposure or lack of progesterone due to ovarian dysfunction can result in endometrial hyperplasia and carcinoma. A key contributor to our understanding of progesterone as a critical regulator for normal uterine function has been the elucidation of progesterone receptor (PR) expression, regulation, and signaling pathways. Several human studies indicate that PR-mediated signaling pathways in the nucleus are associated with progesterone resistance in women with PCOS. The aim of this review is to provide an overview of endometrial progesterone resistance in women with PCOS; to present the PR structure, its different isoforms, and their expression in the endometrium; to illustrate the possible regulation of PR and PR-mediated signaling in progesterone resistance in women with PCOS; and to discuss current clinical treatments for atypical endometrial hyperplasia and endometrial carcinoma in women with PCOS and accompanying progesterone resistance.     

XPC promotes MDM2-mediated degradation of the p53 tumor suppressor

Although ubiquitin receptor Rad23 has been implicated in bringing ubiquitylated p53 to the proteasome, how Rad23 recognizes p53 remains unclear. We demonstrate that XPC, a Rad23-binding protein, regulates p53 turnover. p53 protein in XPC-deficient cells remains ubiquitylated, but its association with the proteasome is drastically reduced, indicating that XPC regulates a postubiquitylation event. Furthermore, we found that XPC participates in the MDM2-mediated p53 degradation pathway via direct interaction with MDM2. XPC W690S pathogenic mutant is specifically defective for MDM2 binding and p53 degradation. p53 is known to become stabilized following UV irradiation but can be rendered unstable by XPC overexpression, underscoring a critical role of XPC in p53 regulation. Elucidation of the proteolytic role of XPC in cancer cells will help to unravel the detailed mechanisms underlying the coordination of DNA repair and proteolysis.