Kisspeptin prevents pregnancy loss by modulating the immune microenvironment at the maternal-fetal interface in recurrent spontaneous abortion.

PROBLEM: Immune factors are crucial in the development of recurrent spontaneous abortion (RSA). This study aimed to investigate whether kisspeptin regulates immune cells at the maternal-fetal interface and whether G protein-coupled receptor 54 (GPR54) is involved in this process, through which it contributes to the pathogenesis of RSA. METHOD OF STUDY: Normal pregnancy (NP) (CBA/J × BALB/c) and RSA (CBA/J × DBA/2) mouse models were established. NP mice received tail vein injections of PBS and KP234 (blocker of kisspeptin receptor), whereas RSA mice received PBS and KP10 (active fragment of kisspeptin). The changes in immune cells in mouse spleen and uterus were assessed using flow cytometry and immunofluorescence. The expression of critical cytokines was examined by flow cytometry, ELISA, Western blotting, and qPCR. Immunofluorescence was employed to detect the coexpression of FOXP3 and GPR54. RESULTS: The findings revealed that the proportion of Treg cells, MDSCs, and M2 macrophages in RSA mice was lower than that in NP mice, but it increased following the tail vein injection of KP10. Conversely, the proportion of these cells was reduced in NP mice after the injection of KP234. However, the trend of γδT cell proportion change is contrary to these cells. Furthermore, FOXP3 and GPR54 were coexpressed in mouse spleen and uterus Treg cells as well as in the human decidua samples. CONCLUSION: Our results suggest that kisspeptin potentially participates in the pathogenesis of RSA by influencing immune cell subsets at the maternal-fetal interface, including Treg cells, MDSC cells, γδT cells, and M2 macrophages.

High GNG13 expression is associated with poor survival in epithelial ovarian cancer and breast cancer.

Recently, change in the GNG13 expression has been shown to result in multiple congenital malformations and sexual reversal, and it was also found in the brain. The aim of this study was to measure the expression levels in epithelial ovarian cancer (EOC) and breast cancer (BC) and assess their value as a potential prognostic marker. The correlation of GNG13 protein expression was detected by immunohistochemistry (IHC) in 119 EOC and 125 BC tissues. Assessment of the associations between GNG13 levels and various clinicopathological features was identified, the relationship between GNG13 and prognosis in BC and EOC patients was analyzed using online resources of Oncomine and Kaplan-Meier plotter. Protein expression levels of GNG13 were both significantly lower in BC and EOC compared with normal tissues (p<0.0001 and p<0.001, respectively). Among the clinicopathological characteristics of BC, tumor grade (p=0.001) and TNM stage (p=0.001) were significantly associated with low expression of GNG13. While in EOC, low expression of GNG13 was significantly related to FIGO stage (p=0.001), presence of metastasis (p=0.001), and CA125 (p=0.001). Our data suggest that GNG13 expression maybe as a new inhibitor, which can strongly inhibit metastasis and partially attenuates tumor growth in EOC and BC.

Discerning drivers and future reduction paths of energy-related CO2 emissions in China: combining EKC with three-layer LMDI.

Recognizing the process and identifying the drivers of energy-related CO2 emissions can provide suggestions for designing carbon emission reduction paths, and then promote the further reduction of carbon emission. In this paper, the carbon emissions in China and its subordinate provinces during 2005-2017 are firstly divided into four stages, named S1, S2, S3, and S4. The results show that China has just entered the S3, and it is impossible to reach the peak of energy-related CO2 emissions with steady economic growth before 2030. Then, three-layer LMDI is utilized to explore the drivers of CO2 emissions, and the impact of urbanization which is separated from the population is considered innovatively. The economic development increases CO2 emissions, while the other drivers have diverse effects, which may be positive or negative, on carbon emissions in different regions. Therefore, four emission reduction paths with provincial characteristics should be followed in the future: (i) three provinces, namely, Ningxia, Shaanxi, and Xinjiang, should optimize multiple basic objectives in parallel; (ii) four provinces, such as Inner Mongolia and Hainan, should optimize the energy structure; (iii) six provinces, such as Jiangxi and Hunan, should optimize the industry structure; and (iv) the other provinces should develop new clean energy according to regional conditions.

High neuropilin and tolloid-like 1 expression associated with metastasis and poor survival in epithelial ovarian cancer via regulation of actin cytoskeleton.

Abnormal expression of neuropilin and tolloid-like 1 (NETO1) has been detected in some human carcinomas. However, the expression of NETO1 and the underlying mechanism in epithelial ovarian cancer (EOC) remain unknown. In this study, we found that a higher NETO1 expression in EOC tissue samples compared to normal ovarian tissue samples was significantly correlated with worse overall survival. Additionally, Cox regression analysis suggested that NETO 1 was independently associated with overall survival. NETO1 overexpression enhanced the EOC cells' migration and invasion capability in vitro via regulation of actin cytoskeleton. Mechanistically, silencing NETO1 reduced the expression of ß-tubulin, F-actin and KIF2A. In conclusion, our results demonstrated the critical role of NETO1 in EOC invasion, and therapies aimed at inhibiting its expression or activity might significantly control EOC growth, invasion and metastatic dissemination.

Stochastic dynamics of an active particle escaping from a potential well.

Active matter systems are driven out of equilibrium by the energy directly supplied at the level of constituent active particles that are self-propelled. We consider a model for an active particle in a potential well, characterized by an active velocity with a constant magnitude but a random orientation subject to white noises. We are interested in the escape of the active particle from the potential well in multiple-dimensional space. We investigate two distinct optimal paths, namely, the shortest arrival-time path and the most probable path, by using the analytical and numerical techniques from optimal control and rare event modeling. In particular, we elucidate the relationship between these optimal paths and the reachable set using the Hamiltonian dynamics for the shortest arrival-time path and the geometric minimum action method for the most probable path, respectively. Numerical results are presented by applying these techniques to a two-dimensional double-well potential.

Dysfunction of Tregs contributes to FGR pathogenesis via regulating Smads signalling pathway.

Fetal growth restriction (FGR) is ranked number two of most common complication of abnormal pregnancy worldwide. The pathogenesis of FGR is complicated due to multiple aetiologies and the exact mechanism for FGR development is currently unknown. T regulatory cells (Tregs) are proven to play central roles in the maintenance of normal pregnancy. Peripheral blood samples of 102 pregnant human were collected analysed using flow cytometry to identify Tregs. We found that reduced Tregs and down-regulation of Foxp3 were observed in peripheral blood of FGR patients. In FGR mouse model, we have found that Tregs were not only reduced in spleen but also in placenta. In vitro, Foxp3 and its transcription regulatory signalling molecules, including P-Smad2, P-Smad3 and Smad4, were diminished as well. Inhibition on Foxp3 expression was partially reversed by overexpression of Smad2 and Smad4. In FGR patients, Western blot results revealed that Foxp3, P-Smad2, P-Smad3 and Smad4 expression was inhibited in placenta. Our preliminary result suggests that maternal-foetal immune tolerance mediated by Tregs plays an essential role in the development of FGR. The inhibited expression of Foxp3 and down-regulated Smad2/Smad3/Smad4 signalling pathway were involved in the FGR pathogenesis. Targeting maternal-foetal immune tolerance through Tregs might represent a novel therapeutic option for FGR.

Myeloid differentiation factor 88 (Myd88) is involved in the innate immunity of black carp (Mylopharyngodon piceus) defense against pathogen infection.

Myeloid differentiation factor 88 (MyD88) is an important transduction protein in the Toll-like receptor signaling pathway. In this study, we identified the cDNA of the MpMyD88 gene in black carp. We found that MpMyD88 was widely distributed in the tissues tested and showed significant immune responses both in vitro and in vivo after stimulation with bacterial and pathogen-associated molecular patterns. After MpMyD88 overexpression/silencing, proinflame-matory cytokines (TNF-α, IFN-α, IL-6, and IL-8) also showed significant up-regulation/down-regulation. Moreover, we found that the antibacterial ability of cells over-expressing MpMyD88 was significantly stronger than that of control cells, while that of silenced MpMyD88 was significantly lower than that in control cells. Besides, we found that the overexpression of MpMyD88 significantly increased the activity of NF-κB. These results indicate that MpMyD88 plays an important role in the innate immune response.

Simplified gentlest ascent dynamics for saddle points in non-gradient systems.

The gentlest ascent dynamics (GAD) [W. E and X. Zhou, Nonlinearity 24, 1831 (2011)] is a time continuous dynamics to efficiently locate saddle points with a given index by coupling the position and direction variables together. These saddle points play important roles in the activated process of randomly perturbed dynamical systems. For index-1 saddle points in non-gradient systems, the GAD requires two direction variables to approximate, respectively, the eigenvectors of the Jacobian matrix and its transposed matrix. In the particular case of gradient systems, the two direction variables are equal to the single minimum mode of the Hessian matrix. In this note, we present a simplified GAD which only needs one direction variable even for non-gradient systems. This new method not only reduces the computational cost for the direction variable by half but also avoids inconvenient transpose operation of the Jacobian matrix. We prove the same convergence property for the simplified GAD as that for the original GAD. The motivation of our simplified GAD is the formal analogy with Hamilton's equation governing the noise-induced exit dynamics. Several non-gradient examples are presented to demonstrate our method, including a two dimensional model and the Allen-Cahn equation in the presence of shear flow.

Combined treatment of neurotrophin-3 gene and neural stem cells is ameliorative to behavior recovery of Parkinson's disease rat model.

Neural stem cell transplantation therapy was developed for replacing lost or damaged neural cells for the neurodegenerative disease, including Parkinson's disease (PD), in which dopaminergic neuron cells are lost. The growth factor, neurotrophin-3(NT-3), has been shown to promote neuroregeneration, differentiation and migration during brain development. In this report, we construct rat neural stem cells that express neurotrophin-3 endogenously (rNSC-NT3) and transplant them into 6-hydroxydopamine (6-OHDA)-treated Parkinsonian rats. Molecular approaches including quantitative real time PCR, Western blot and immunocytochemistry were used to identify the expression of NT-3 and the differentiation of planted cells. Behavioral recover was also tested. The result indicated that combined treatment of neurotrophin-3 gene and neural stem cells had a functional impact on reversing the main symptoms of the Parkinson's disease that significantly reduced apomorphine-induced rotational asymmetry and improved spatial learning ability. The rNSCs-NT3 is able to differentiate into dompaminergic neuron in the ventral tegmental area (VTA) and the medial forebrain bundle (MFB), and migrated around the lesion site. Endogenous expressed NT-3 exerts induction and trophic effects on neural stem cells. The rNSCs-NT3 showed higher activity than the rNSCs in regenerating tyrosine hydroxylase positive cell numbers and migrating distance, behavior improving in this dopa-deficit rat model. These findings suggest that the neural stem cells expressed NT-3 endogenously would be a better graft candidate for the treatment of Parkinson's disease.

Combined treatment of neurotrophin-3 gene and neural stem cells is propitious to functional recovery after spinal cord injury.

We present an insight of the effects of combination therapy with neurotrophin-3 and neural stem cell on functional recovery after spinal cord injury (SCI). Total RNA was extracted from neural stem cell line C17.2 and reversed transcribed into cDNA. Neurotrophin-3 (NT-3) gene was amplified by PCR and subcloned into plasmid to construct an expression vector pNT-3. A positive clone containing pNT-3, named SHN2, was obtained and used for transplantation. Thirty adult mice received mechanical injury at the T8 vertebra level. Cell survival, NT-3 gene expression, and functional recovery were observed through X-Gal staining, RT-PCR, and open field locomotion, respectively. The results show that NT-3 gene comprising 777 bp nucleotides was cloned and a more than twofold expression was detected when transfected into neural stem cell line C17.2. Quantitative analysis of cellular density revealed a significant increase in SHN2 compared to the control cells (p < 0.01). Thirty days after transplantation, SHN2 showed significant increase near the lesion site. Furthermore, the functional recovery indicated an active effect by detecting Basso-Beattie-Bresnahan (BBB) locomotor rating scale (p < 0.01). In conclusion, combined treatment of neural stem cells and NT-3 gene can facilitate functional recovery. It offers an effective approach to treat SCI.

Combination of bFGF, heparin and laminin induce the generation of dopaminergic neurons from rat neural stem cells both in vitro and in vivo.

Neural stem cells (NSCs) are self-renewing and pluripotent, which can differentiate into neurons, astrocytes and oligodendrocytes. Due to these properties, NSCs are supposed to be an ideal candidate to clinical purpose while research on cell replacement therapy to treat neural diseases has been widely investigated recently. In this article, we demonstrated a new and efficient method to induce the generation of proliferative dopaminergic neurons from rat NSCs in the presence of bFGF, heparin and laminin both in vitro and in vivo. These cells were testified to survive in the grafted 6-Hydroxy-Dopamine (6-OHDA) lesioned rat for at least 1 month. More importantly, migration to close host tissue was observed on day 30 post-transplantation. In this regard, we anticipated that this technology may advance stem cell-based therapy to replace lost neurons in neural injury or neurodegenerative disorders.