Microarray Expression Profiles of lncRNAs and mRNAs in Postoperative Cognitive Dysfunction

Postoperative cognitive dysfunction (POCD) is serious disorder in the central nervous system common in aged patients after anesthesia. Although its clinical symptoms are well recognized, however, the molecular etiology of the POCD remains unrevealed. Similarly, neither gold standard molecular diagnosis nor effective treatment is available for POCD until the present. Therefore, we aimed to explore the molecular mechanism of this disorder through investigating lncRNAs and mRNAs associated with POCD human patients and investigate their underlying regulatory pathways. In this study, we recruited 200 patients requiring hip or knee replacement surgery. Their neurological functions were assessed at two time points, 1 day before the surgery and 30 days post-surgery. In parallel, serum samples were collected from the participants to analyze lncRNAs and mRNAs differential expression profile between POCD and non-POCD patients using microarray analysis. To further investigate the role differentially expressed mRNA and lncRNAs, Gene Ontology (GO), pathway analyses on mRNAs and lncRNA-mRNA interaction network were performed. As a result, 68 lncRNAs and 115 mRNAs were dysregulated in the POCD group compared to non-POCD group. Among them, the top 10 upregulated lncRNAs and 10 downregulated lncRNAs were listed for enrichment analysis. Interestingly, we found that these lncRNA and mRNA are involved in biological process, molecular function, and cellular component in addition to various signaling pathways, suggesting that the pathogenesis of POCD involves lncRNAs and mRNAs differential expression. Consequently, the genetic dysregulation between the non-POCD and POCD patients participates in the occurrence and development of POCD, and could be served as diagnostic biomarkers and drug targets for POCD treatment

A Novel Role of VEGFC in Cerebral Ischemia With Lung Injury

Cerebral ischemia (CI) is a severe brain injury resulting in a variety of motor impairments combined with secondary injury in remote organs, especially the lung. This condition occurs due to insufficient blood supply to the brain during infancy. However, it has a molecular linkage that needs to be thoroughly covered. Here, we report on the role of vascular endothelial growth factor C (VEGFC) in lung injury induced by CI. The middle cerebral artery occlusion (MCAO) was depended to establish the animal model of CI. Rats were used and brain ischemia was confirmed through TTC staining. Serum was used for protein chip analysis to study the proteomic interaction. Immunohistochemistry analyses were used to quantify and locate the VEGFC in the lung and brain. The role of VEGFC was detected by siVEGFC technology in SY5Y, HUCEV, and A549 cell lines, under normal and oxygen glucose deprivation (OGD) conditions in vitro. As a result, the TTC staining demonstrated that the model of brain ischemia was successfully established, and MPO experiments reported that lung damage was induced in MCAO rats. VEGFC levels were up-regulated in serum. On the other hand, immunohistochemistry showed that VEGFC increased significantly in the cytoplasm of neurons, the endothelium of small trachea and the lung cells of CI animals. On a functional level, siVEGFC effectively inhibited the proliferation of SY5Y cells and decreased the viability of HUVEC cells in normal cell lines. But under OGD conditions, siVEGFC decreased the growth of HUVEC and increased the viability of A549 cells, while no effect was noticed on SYSY cells. Therefore, we confirmed the different role of VEGFC played in neurons and lung cells in cerebral ischemia-reperfusion injury. These findings may contribute to the understanding the molecular linkage of brain ischemia and lung injury, which therefore provides a new idea for the therapeutic approach to cerebral ischemia-reperfusion

A direct and non-invasive method for kidney delivery of therapeutics in mice

Kidney is a vital organ that maintains the homeostasis in terms of acid-balance, toxin filtration and blood pressure control. Kidney malfunction can be fatal and the renal research administers testing pharmaceutical agents or stem cells in rodents to study their therapeutic efficacy. However, targeted delivery of agents into mice kidney is strenuous and may require laparotomy. Here we present a direct delivery method for cell transplantation or drug injection into the mice kidney. The method is simple and can be performed non-invasively with avoidance of surgical intervention on the animals. Nevertheless, this method serves as an efficient method for in vivo drug delivery or engraftment studies for renal research. • Direct delivery into the kidney. • Non-invasive method. Method name: Direction injection into mice kidney, Keywords: Kidney, Drugs, Delivery, Surgery, Injectio

Facial vein injection of human cells in severe combined immunodeficiency (SCID) neonatal mice

Intravenous injection is a standard procedure for delivering human stem cells and therapeutic agents. Currently, genetically modified severe combined immunodeficiency (SCID) mice are used for engraftment studies using human cells. SCID neonates have better integration and survivability of human cells compared to adult SCID mice, as their immune system will not be developed in the first few days after birth. However, intravenous injections in neonates are difficult. This protocol describes a reliable and reproducible method for injecting cells into the facial vein of P3/P4 (3 or 4 days post-birth) SCID neonates to study their engraftment. The injection was safe and well tolerated by the pups. Post-injection analysis revealed the distribution of tagged cells in different organs. Results suggest that this new method can serve as a pre-analysis for transplantation studies using human stem cells before in vivo animal model testing. Method name: Facial vein injection in P3/P4 pups, Keywords: Neonates, Facial vein, Intravenous injection, Stem cells, Deliver

Characterization of Urine Stem Cell-Derived Extracellular Vesicles Reveals B Cell Stimulating Cargo

Elucidation of the biological functions of extracellular vesicles (EVs) and their potential roles in physiological and pathological processes is an expanding field of research. In this study, we characterized USC–derived EVs and studied their capacity to modulate the human immune response in vitro. We found that the USC–derived EVs are a heterogeneous population, ranging in size from that of micro–vesicles (150 nm–1 μm) down to that of exosomes (60–150 nm). Regarding their immunomodulatory functions, we found that upon isolation, the EVs (60–150 nm) induced B cell proliferation and IgM antibody secretion. Analysis of the EV contents unexpectedly revealed the presence of BAFF, APRIL, IL–6, and CD40L, all known to play a central role in B cell stimulation, differentiation, and humoral immunity. In regard to their effect on T cell functions, they resembled the function of mesenchymal stem cell (MSC)–derived EVs previously described, suppressing T cell response to activation. The finding that USC–derived EVs transport a potent bioactive cargo opens the door to a novel therapeutic avenue for boosting B cell responses in immunodeficiency or cancer