Genetic difference between two Schistosoma japonicum isolates with contrasting cercarial shedding patterns revealed by whole genome sequencing.

Schistosoma japonicum is one of the major infectious agents of human schistosomiasis, mainly endemic in China and the Philippines. We have previously reported the finding of two schistosome isolates, each with a different cercarial emergence pattern adapted to their different hosts. However, there are currently no whole-genome sequencing studies to investigate the underlining genetics of the adaptive traits. We sampled schistosomes in 2013 and 2020 from a hilly area Shitai (ST) and a marshland area Hexian (HX) of Anhui, China. Ten to 15 male or female adult worms from each site/year were sent for whole genome sequencing. Genetics were analyzed, and selection signals along genomes were detected. Gene enrichment analysis was performed for the genome regions under selection. The results revealed considerable genetic differentiation between the two isolates. The genome "windows" affected by natural selection were fewer in ST (64 windows containing 78 genes) than in HX (318 windows containing 276 genes). Twelve significantly enriched genes were identified in ST, but none in HX. These genes were mainly related to specific DNA binding and intercellular signaling transduction. Some functional region changes identified along the genome of the hilly schistosome may be related to its unique late afternoon cercarial emergence.

Title: Différence génétique entre deux isolats de Schistosoma japonicum présentant des schémas contrastés d'émergence de cercaires, révélés par séquençage du génome entier. Abstract: Schistosoma japonicum est l'un des principaux agents infectieux de la schistosomiase humaine, principalement endémique en Chine et aux Philippines. Nous avons précédemment rapporté la découverte de deux isolats de schistosomes, chacun présentant un schéma différent d'émergence de cercaires, adapté à leurs différents hôtes. Cependant, il n'existe actuellement aucune étude de séquençage du génome entier pour comprendre la génétique sous-jacente aux traits adaptatifs. Nous avons échantillonné des schistosomes en 2013 et 2020 dans une zone de collines de Shitai (ST) et une zone marécageuse de Hexian (HX) de l'Anhui, en Chine. Dix à 15 vers adultes mâles ou femelles de chaque site/année ont été envoyés pour séquençage du génome entier. La génétique a été analysée et des signaux de sélection le long des génomes ont été détectés. Une analyse d'enrichissement génétique a été réalisée pour les régions du génome sélectionnées. Les résultats ont révélé une différenciation génétique considérable entre deux isolats. Les « fenêtres ¼ du génome affectées par la sélection naturelle étaient moins nombreuses dans ST (64 fenêtres contenant 78 gènes) que dans HX (318 fenêtres contenant 276 gènes). Douze gènes significativement enrichis ont été identifiés dans ST mais aucun dans HX. Ces gènes étaient principalement liés à la liaison spécifique à l'ADN et à la transduction de la signalisation intercellulaire. Certains changements de régions fonctionnelles identifiés le long du génome du schistosome des collines peuvent être liés à son émergence cercarienne exceptionnelle en fin d'après-midi.

Nampt promotes osteogenic differentiation and lipopolysaccharide-induced interleukin-6 secretion in osteoblastic MC3T3-E1 cells.

The Nicotinamide phosphoribosyltransferase (Nampt)-NAD-Sirt1 pathway modulates processes involved in the pathogenesis of multiple diseases by influencing inflammation. This study aimed to explore the effect of Nampt in osteogenic differentiation and inflammatory response of osteoblastic MC3T3-E1 cells. We developed an in vitro model of lipopolysaccharide (LPS)-induced inflammation and showed that Nampt and Sirt1 were significantly upregulated in LPS-treated MC3T3-E1 cells. LPS induced secretion of the proinflammatory cytokine interleukin-6 (IL-6) and attenuated osteogenic differentiation. Then we transfected cells with adenoviruses to knock down or over express Nampt. Nampt promoted the expression of IL-6, TAK1 and phospho-NF-κB p65 after LPS treatment. Overexpression of Nampt overrode the effect of LPS and rescued LPS-induced inhibition on osteogenic differentiation. FK866, a Nampt inhibitor, had the same inhibitory effect as Nampt knockdown. In addition, Sirt1 suppression by EX527 decreased IL-6 secretion and NF-κB activation without changing the level of Nampt. EX527 also decreased osteogenic differentiation. Incubation with NMN or SRT 1720 also counteract the inhibitory effect of LPS and rescued osteoblast differentiation. Therefore, we demonstrated that Nampt acted both in promoting osteoblast differentiation and in enhancing inflammatory response, mediated by Sirt1 in MC3T3-E1 cells.

SDF1/CXCR4 axis facilitates the angiogenesis via activating the PI3K/AKT pathway in degenerated discs.

Symptomatic degenerative disc disease (DDD) is considered the leading cause of chronic lower back pain (LBP). As one of the main features of intervertebral disc degeneration (IDD), vascular ingrowth plays a crucial role in the progression of LBP. Stromal cell­derived factor 1 (SDF1) and its receptor C­X­C receptor 4 (CXCR4) were reported to be overexpressed in the degenerated intervertebral discs, suggesting that they may be involved in the pathogenesis of IDD. Moreover, SDF1 has been identified to induce neovascularization in rheumatoid arthritis disease. However, the roles of the SDF1/CXCR4 axis in the neovascularization of IDD remain unclear. Therefore, the objective of the present study was to elucidate whether the SDF1/CXCR4 axis takes part in neovascularization in degenerated intervertebral discs and its underlying mechanisms. Adenovirus infection was used to upregulate SDF1 expression in primary nucleus pulposus cells (NPCs). The effects of SDF1 on the proliferation and angiogenesis of vascular endothelial cells (VECs) were assessed by Cell Counting Kit­8 and tube formation assays after VECs were treated with the supernatants derived from SDF1 overexpressed or not treated NPCs. Transwell chambers using the supernatants from NPCs as chemokines were applied to assess VEC migration and invasion. AMD3100, MK­2206 and SF1670 were used to antagonize CXCR4, AKT serine/threonine kinase 1 (AKT) and phosphatase and tensin homolog (PTEN) in VECs. The results revealed that SDF1 overexpression significantly increased the ratio of phosphorylated AKT to AKT and decreased PTEN expression in NPCs, as well as enhanced the proliferation, migration, invasion and angiogenesis abilities of VECs. However, these effects induced by SDF1 overexpression in NPCs were all reversed when VECs were pretreated with AMD3100 or MK­2206, whereas enhanced by SF1670 treatment. Collectively, the present study indicated that enhancement of the SDF1/CXCR4 axis in NPCs can significantly accelerate angiogenesis by regulating the PTEN/phosphatidylinositol­3­kinase/AKT pathway.

SDF1/CXCR4 axis plays a role in angiogenesis during the degeneration of intervertebral discs.

Low back pain (LBP) is a ubiquitous disease affecting quality of life. The ingrowth of new blood vessels is an important pathological feature of LBP, but its underlying mechanisms are poorly understood. The present study aimed to investigate the influence and relative mechanism of stromal cell derived factor 1 (SDF1) on the angiogenesis of degenerated intervertebral discs. The expression of SDF1 in nucleus pulposus cells (NPCs) was upregulated and downregulated by virus transfection, and the NPCs were allocated to either the downregulation (Down), degeneration (D) or upregulation (Up) group according to the expression of SDF1. The different groups of NPCs or NPC conditioned media were co­cultured with vascular endothelial cells (VECs) under different conditions. A Cell Counting Kit­8 (CCK­8) assay, a Transwell migration assay and a tube formation assay were conducted to evaluate the influence on angiogenesis. The results showed that SDF1 was significantly up­ and downregulated in the Up and Down groups, respectively. Each group of NPCs or their conditioned medium was co­cultured with VECs; the CCK­8, Transwell migration and tube formation assays showed that cell viability, chemotactic migration and the tube formation ability of VECs increased with the rise in SDF1. The aforementioned results were significantly different between each group. After adding the CXCR4 inhibitor, AMD3100, the viability, migration and tube formation of VECs were suppressed in the D and Up groups, and there was a significant difference compared with the prior to the addition of the inhibitor, while there was a declining tendency in the Down group and no significant difference following addition of the inhibitor. The results demonstrated that SDF1 is expressed in human NPCs, and the SDF1/CXCR4 axis can influence the viability, migration and tube formation of VECs and may play an important role in the angiogenesis of human degenerated discs.

SDF1/CXCR7 Signaling Axis Participates in Angiogenesis in Degenerated Discs via the PI3K/AKT Pathway.

Degenerative disc disease (DDD) is the main cause of low back pain, and the ingrowth of new blood vessels is one of its pathological features. The stromal cell-derived factor 1 (SDF1)/CXCR7 signaling axis plays a role in these physiological and pathological activities. The aims of this study were to explore whether this signaling axis participates in the angiogenesis of degenerated intervertebral discs (IVDs) and to define its underlying mechanism. In this study, we cocultured human nucleus pulposus cells (NPCs) and vascular endothelial cells (VECs) and regulated the expression of SDF1/CXCR7 to investigate the effect of VEC angiogenesis by NPCs. The results revealed that angiogenesis was enhanced with increased SDF1 and that angiogenesis was weakened with the inhibition of CXCR7. We found that PI3K/AKT was involved in the downstream pathway in the coculture. VEC angiogenesis induction by NPCs was enhanced with an increase in pAKT or a decrease in PTEN. We conclude that the SDF1/CXCR7 signaling axis plays a role in the angiogenesis of degenerated IVD through the PI3K/AKT pathway.

BMP9 Promotes the Extracellular Matrix of Nucleus Pulposus Cells via Inhibition of the Notch Signaling Pathway.

Intervertebral disk degeneration (IDD) is a common disease that is caused by degeneration of the nucleus pulposus (NP). One goal in the treatment of IDD is delaying or reversing the degeneration of NP via the transformation of exogenous genes. This study first investigated the role of BMP9 in the extracellular matrix (ECM) of nucleus pulposus cells (NPCs) and its mechanism. We found that BMP9 promotes the expression of ECM in NPCs, and the key molecules of Notch signaling, namely, NICD-1, hes and hey, and it was significantly altered in BMP9-transfected NPCs, which suggests that BMP9 may regulate the ECM via the Notch signaling pathway. We verified the expression of Notch ligands and receptors in NPCs infected with Ad-BMP9 and demonstrated a significant decrease in DLL1 and Notch1; then, NPCs were transfected with Ad-dnNotch1, Ad-Jagged1, and Ad-DLL1, and different multiple groups were established to further identify the ligands or receptors that affected ECM expression. The results demonstrated that Ad-dnNotch1, Jagged1 and DLL1 inhibited ECM expression, and dnNotch1 promoted expression. Therefore, we demonstrated that BMP9 promoted the expression of ECM in NPCs via inhibition of Notch1 and DLL1. This study provides a possible method for IDD treatment.

Resveratrol ameliorates autophagic flux to promote functional recovery in rats after spinal cord injury.

Resveratrol is known to improve functional recovery after spinal cord injury, but the exact mechanism involved is yet unclear. The aim of this study was to clarify whether resveratrol can exert neuroprotective effects via activating neuronal autophagic flux, in view of the underlying role of the autophagic flux mediated by resveratrol on neuronal apoptosis after spinal cord injury, and identify the role of the liver kinase B1(LKB1)/adenosine monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR)/ p70 ribosomal protein S6 kinase (p70s6k) signal pathway in the autophagic flux mediated by resveratrol. The results obtained strongly indicate that resveratrol improved functional recovery in Sprague-Dawley rats after acute spinal cord injury, preserved their motor neurons, alleviated the neuronal apoptosis, and ameliorated neuronal autophagic flux. After blocking the autophagic flux, the neuroprotective effects of resveratrol were eliminated. Furthermore, it was proved that resveratrol can activate the LKB1/AMPK/mTOR/p70s6k pathway in vivo and in vitro, and the LKB1/AMPK/mTOR/p70s6k pathway plays a vital role in activating the autophagic flux mediated by resveratrol in PC12 cells. Thus, resveratrol enables to ameliorate neuronal autophagic flux via the LKB1/AMPK/mTOR/p70s6k pathway to alleviate apoptosis, and finally ameliorating functional recovery after acute SCI in SD rats.

Identifying dysregulated pathways in postmenopausal osteoporosis through investigation of crosstalk between pathways.

The present study aimed to identify potential dysregulated pathways to further reveal the molecular mechanisms of postmenopausal osteoporosis (PMOP) based on pathway­interaction network (PIN) analysis, which considers crosstalk between pathways. Protein­protein interaction (PPI) data and pathway information were derived from STRING and Reactome Pathway databases, respectively. According to the gene expression profiles, pathway data and PPI information, a PIN was constructed with each node representing a biological pathway. Principal component analysis was used to compute the pathway activity for each pathway, and the seed pathway was selected. Subsequently, dysregulated pathways were extracted from the PIN based on the seed pathway and the increased classification accuracy, which was measured using the area under the curve (AUC) index according to 5­fold cross validation. A PIN comprising 2,725 interactions was constructed, which was used to detect dysregulated pathways. Notably, the 'mitotic prometaphase' pathway was selected and defined as a seed pathway. Starting with the seed pathway, network­based analysis successfully identified one pathway set for PMOP comprising eight dysregulated pathways (such as mitotic prometaphase, resolution of sister chromatid cohesion, mRNA splicing and mRNA splicing­major) with an AUC score of 0.85, which may provide potential biomarkers for targeted therapy for PMOP.