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Abstract Objective Hypoxia can promote stem cell proliferation and migration through HIF-1α. Hypoxia can regulate cellular endoplasmic reticulum (ER) stress. Some studies have reported the relationship among hypoxia, HIF-α, and ER stress, however, while little is known about HIF-α and ER stress in ADSCs under hypoxic conditions. The purpose of the study was to investigate the role and relationship of hypoxic conditions, HIF-1α and ER stress in regulating adipose mesenchymal stem cells (ADSCs) proliferation, migration, and NPC-like differentiation. Method ADSCs were pretreated with hypoxia, HIF-1α gene transfection, and HIF-1α gene silence. The ADSCs proliferation, migration, and NPC-like differentiation were assessed. The expression of HIF-1α in ADSCs was regulated; then, the changes of ER stress level in ADSCs were observed to investigate the relationship between ER stress and HIF-1α in ADSCs under hypoxic conditions. Result The cell proliferation and migration assay results show that hypoxia and HIF-1α overexpression can significantly increase the ADSCs proliferation and migration, while HIF-1α inhibition can significantly decrease the ADSCs proliferation and migration. The HIF-1α and co-cultured with NPCs played an important role in the directional differentiation of ADSCs into NPCs. The hypoxia-regulated ER stress in ADSCs through the HIF-1α pathway, thereby regulating the cellular state of ADSCs, was also observed. Conclusion Hypoxia and HIF-1α play important roles in proliferation, migration, and NPC-like differentiation of ADSCs. This study provides preliminary evidence that HIF-1α-regulated ER stress thus affects ADSCs proliferation, migration, and differentiation. Therefore, HIF-1α and ER may serve as key points to improve the efficacy of ADSCs in treating disc degeneration.

Objective: Hypoxia can promote stem cell proliferation and migration through HIF-1α. Hypoxia can regulate cellular endoplasmic reticulum (ER) stress. Some studies have reported the relationship among hypoxia, HIF-α, and ER stress, however, while little is known about HIF-α and ER stress in ADSCs under hypoxic conditions. The purpose of the study was to investigate the role and relationship of hypoxic conditions, HIF-1α and ER stress in regulating adipose mesenchymal stem cells (ADSCs) proliferation, migration, and NPC-like differentiation. Method: ADSCs were pretreated with hypoxia, HIF-1α gene transfection, and HIF-1α gene silence. The ADSCs proliferation, migration, and NPC-like differentiation were assessed. The expression of HIF-1α in ADSCs was regulated; then, the changes of ER stress level in ADSCs were observed to investigate the relationship between ER stress and HIF-1α in ADSCs under hypoxic conditions. Result: The cell proliferation and migration assay results show that hypoxia and HIF-1α overexpression can significantly increase the ADSCs proliferation and migration, while HIF-1α inhibition can significantly decrease the ADSCs proliferation and migration. The HIF-1α and co-cultured with NPCs played an important role in the directional differentiation of ADSCs into NPCs. The hypoxia-regulated ER stress in ADSCs through the HIF-1α pathway, thereby regulating the cellular state of ADSCs, was also observed. Conclusion: Hypoxia and HIF-1α play important roles in proliferation, migration, and NPC-like differentiation of ADSCs. This study provides preliminary evidence that HIF-1α-regulated ER stress thus affects ADSCs proliferation, migration, and differentiation. Therefore, HIF-1α and ER may serve as key points to improve the efficacy of ADSCs in treating disc degeneration. [ABSTRACT FROM AUTHOR]

BACKGROUND: Stem cells have the potential of self-renewal and pluripotent directional differentiation, which are necessary for the body to maintain physiological functions, and play an important role in the occurrence and development of various diseases, disease treatment and organ transplantation. Recent studies have found that m6A methylation modification can regulate the biological characteristics of stem cells and play an important role in various diseases. OBJECTIVE: To mainly summarize the latest research progress of biological process of m6A methylation modification of stem cells, including self-renewal, directional differentiation, and immunological characteristics. METHODS: Related articles published from 1955 to 2022 were retrieve from PubMed, Web of Science and CNKI databases. The keywords were “stem cell, m6A, RNA methylation, biological characteristics, self-renewal, proliferation, differentiation, immunization” in English and Chinese. Ultimately, we included 88 articles for review. RESULTS AND CONCLUSION: m6A, as the epigenetic modification with the highest abundance in eukaryotic mRNA, is also the focus of current epigenetic research. It is composed of methyltransferase, demethylase and methyl-binding domain protein, and has become an important transcriptome marker. The conclusions of m6A methylation regulate the biological characteristics as following: (1) m6A methylation modification can be adjusted by related enzymes target transformation, proliferation, cell cycle-related molecular transcription after translation, the expression of key enzymes in the process of DNA replication, to a particular cell differentiation, immune checkpoint molecular expression and their sensitivity to radiation and chemotherapy drugs, and in the treatment of malignant disease, delay condition, improve the prognosis of playing a key role. (2) m6A-related enzymes are closely related to the biological characteristics of stem cell proliferation, self-renewal, directed differentiation and immune regulation, and can be used as a target for drug therapy. (3) The study of m6A methylation on human diseases is still in the basic research stage, and the mechanism of m6A modification in some malignant diseases has not been thoroughly explored. With the development of epigenetics, RNA m6A modification is expected to achieve new breakthroughs in the treatment of malignant diseases and clinical transformation. [ABSTRACT FROM AUTHOR]

Objective: This study is to research and develop a simple and rapid method to directly differentiate the induced pluripotent stem cells (iPS) into functional hepatocytes in vitro. Methods: According to the development of normal hepatocytes in vitro, a simplified method was designed for differentiation of iPS cells into endodermal cells. The phenotypes of endodermal cells was identified by qPCR and flow cytometry, and then these cells were further differentiated into hepatocyte-like cells. The characteristics and functions of hepatocytes were identified by qPCR, ELISA and immunofluorescence. Results: The results showed that the mRNA/protein expression of OCT4 and NANOG were decreased significantly, and the mRNA/protein expression of endodermal cell-related genes, CXCR4, FOXA2 and HNF4A,were increased significantly at day 7 after iPS cell induction. Hepatocyte specific marker genes ALB, TDO2, RBP4, G6PC and hepatocyte drug enzyme genes CYPs were significantly upregulated at day 15, and high levels of albumin and urea were produced simultaneously. Positive PAS glycogen staining and active uptake and release of indocyanine green were observed in the induced hepatocytes. It was confirmed that the hepatocyte-like cells had the partial functions of primary hepatocytes. Conclusion: This easy and efficient induction method can be used in differentiation of iPS cells into functional hepatocytes by mimicking the process of liver organogenesis in vitro. This study provides the possibility for massive production of iPS-derived hepatocytes and their applications in cellular therapy and drug screening models. [ABSTRACT FROM AUTHOR]

A group of cells in cartilage tissue with stem cell characteristics may be ideal seed cells for cartilage defect repair. In previous studies, stem/progenitor cells from articular cartilage have been isolated and identified in humans, horses, and bovines, but rarely in ducks. We isolated and cultured cartilage stem/progenitor cells (CPSCs) from the articular cartilage of 14-day-old Peking duck embryos using the fibronectin adhesion method. The morphology of CPSCs was observed under an inverted microscope. Based on the growth curve test, we found that the growth mode of the cells was “S” type growth. The clonogenic assay was performed to determine the clonogenic efficiency CPSCs. The expression of cell surface markers was detected by immunofluorescence, flow cytometry, and RT-PCR, and it was found that the CPSCs positively expressed FGFR-3, collagen type II, CD44, CD73, CD90, vimentin, Notch1, CD105, and CD106, but negatively expressed the haematopoietic stem cell surface marker CD34. Furthermore, CPSCs were successfully induced to differentiate into adipocytes, osteoblasts, and chondrocytes, showing that they have an in vitro differentiation ability. These findings provide a theoretical and experimental basis for cell transplantation therapy in tissue engineering. [ABSTRACT FROM AUTHOR]

Neural precursor cells (NPCs) generate new neurons to supplement neuronal loss as well as to repair damaged neural circuits. Therefore, NPCs have potential applications in a variety of neurological diseases, such as spinal cord injury, traumatic brain injury, and glaucoma. Specifically, improving NPCs proliferation and manipulating their differentiated cell types can be a beneficial therapy for a variety of these diseases. ADT‐OH is a slow‐releasing organic H2S donor that produces a slow and continuous release of H2S to maintain normal brain functions. In this study, we aimed to explore the effect of ADT‐OH on NPCs. Our results demonstrated that ADT‐OH promotes self‐renewal and antiapoptosis ability of cultured NPCs. Additionally, it facilitates more NPCs to differentiate into neurons and oligodendrocytes, while inhibiting their differentiation into astrocytes. Furthermore, it enhances axonal growth. Moreover, we discovered that the mRNA and protein expression of β‐catenin, TCF7L2, c‐Myc, Ngn1, and Ngn2, which are key genes that regulate NPCs self‐renewal and differentiation, were increased in the presence of ADT‐OH. Altogether, these results indicate that ADT‐OH may be a promising drug to regulate the neurogenesis of NPCs, and needs to be studied in the future for clinical application potential. [ABSTRACT FROM AUTHOR]

Progressive loss of dopaminergic neurons is the primary pathological feature of Parkinson's disease (PD), which causes severe cognitive and motor dysfunction. In this context, transplantation of neural stem cells (NSCs) is the most promising approach to supply new dopaminergic neurons for the treatment of PD, which faces the challenge of precisely controlling the differentiation direction of NSCs. Herein, we report the efficient differentiation of NSCs into neurons boosted by the joint use of Cu 2- x Se nanoparticles (CS NPs) with the second near-infrared (NIR-II) laser irradiation. We reveal the mechanism of neuronal differentiation of NSCs and find that the voltage-gated calcium channels (VGCC) of NSCs can be opened under the stimulation of CS NPs and NIR-II laser irradiation to trigger Ca2+/CaMK/CREB/c-Fos signaling pathway, which promotes the neural differentiation and survival. More importantly, when a mixture of CS NPs and NSCs was injected into the damaged striatum of 6-hydroxydopamine(6-OHDA)-induced PD mice and then irradiated with an external 1064 nm NIR-II laser, the NSCs were efficiently differentiated into dopaminergic neurons to effectively repair the damaged neural circuits in the nigra-striatum and ameliorate the motor disorders and anxiety of PD mice. These findings demonstrate the feasibility and importance of precise modulation of directional differentiation of NSCs into matured functional neurons in vivo and in vitro through advanced functional nanoparticles and nanotechnology, which offers a promising approach for the treatment of PD and other neurodegenerative diseases. [Display omitted] • Cu 2-x Se nanoparticles (CS NPs) and NIR Ⅱ laser irradiation promote neural stem cells (NSCs) differentiation into neurons. • The voltage-gated calcium channels (VGCC) of NSCs can be opened to trigger Ca2+/CaMK/CREB/c-Fos signaling pathway. • The transplanted NSCs assited with CS NPs and 1064 nm NIR-Ⅱ laser ameliorate the symptoms of PD mice. [ABSTRACT FROM AUTHOR]

Interleukin 17 (IL-17) and its main producer, T cell receptor γδ cells, have neurotoxic effects in the pathogenesis of intracerebral hemorrhage (ICH), aggravating brain injuries. To investigate the correlation between IL-17 and ICH, we dynamically screened serum IL-17 concentrations using enzyme-linked immunosorbent assay and explored the clinical values of IL-17 in ICH patients. There was a significant negative correlation between serum IL-17 level and neurological recovery status in ICH patients (r = –0.498, P < 0.01). To study the neurotoxic role of IL-17, C57BL/6 mice were used to establish an ICH model by injecting autologous blood into the caudate nucleus. Subsequently, the mice were treated with mouse neural stem cells (NSCs) and/or IL-17 neutralizing antibody for 72 hours. Flow cytometry, brain water content detection, Nissl staining, and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling results indicated that NSC transplantation significantly reduced IL-17 expression in peri-hematoma tissue, but there was no difference in T cell receptor γδ cells. Compared with the ICH group, there were fewer apoptotic bodies and more Nissl bodies in the ICH + NSC group and the ICH + NSC + IL-17 group. To investigate the potential effect of IL-17 on directional differentiation of NSCs, we cultured mouse NSCs (NE-4C) alone or co-cultured them with T cell receptor γδ cells, which were isolated from mouse peripheral blood mononuclear cells, for 7 days. The results of western blot assays revealed that IL-17 secreted by T cell receptor γδ cells reduced the differentiation of NSCs into astrocytes and neurons, while IL-17 neutralization relieved the inhibition of directional differentiation into astrocytes rather than neurons. In conclusion, serum IL-17 levels were elevated in the early stage of ICH and were negatively correlated with outcome in ICH patients. Animal experiments and cytological investigations therefore demonstrated that IL-17 probably has neurotoxic roles in ICH because of its inhibitory effects on the directional differentiation of NSCs. The application of IL-17 neutralizing antibody may promote the directional differentiation of NSCs into astrocytes. This study was approved by the Clinical Research Ethics Committee of Anhui Medical University of China (For human study: Approval No. 20170135) in December 2016. All animal handling and experimentation were reviewed and approved by the Institutional Animal Care and Use Committee of Anhui Medical University (approval No. 20180248) in December 2017.

To establish an effective method for acquiring bone marrow-derived dendritic cells (DCs) from BALB/C mice in vitro and to establish a reservoir of DC precursor cells. Methods CD117+ hematopoietic stem cells (HSCs) were isolated and purified from bone marrow of BALB/C mice by immunomagnetic beads separation system (MACS), and then amplified in vitro with mouse stem cell factor (SCF) and interleukin-3 (IL-3). HSC was induced to differentiate into DCs by adding granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and IL-4. Different cytokines (tumor necrosis factor-alpha or IL-10) were added to control the maturity of dendritic cells. Then the morphology (electron microscopy), surface molecular markers (FACS method) and cytokine secretion level (ELISA method) were identified. Results ① The purity of CD117 + HSC isolated and purified by MACS system was over 95%. ② SCF plus IL-3 could effectively stimulate HSC amplification. ③ The morphology of mature DC (mDC) and immature DC (imDC) was significantly different under light and scanning electron microscopy. ④ In the expressions of surface markers CD40, CD80, CD86, I-A/I-E, there were significant differences between imDC group and mDC group (P<0.01). ⑤ After LPS stimulation, the secretion of IL-12 in imDC group did not change significantly (P=0.064), while the secretion of IL-12 in mDC group increased significantly (P=0.009). LPS and TNF-α had a synergistic effect in stimulating DC maturation. Conclusion Specific combinations of cytokines can effectively induce the differentiation of bone marrow HSCs into DCs in BALB/C mice, and can control the maturity of DCs. This study makes it possible to use gene modified dendritic cells in GD immunotherapy. [ABSTRACT FROM AUTHOR]

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A generalized framework for numerical differentiation (ND) is proposed for constructing a finite impulse response (FIR) filter in closed form. The framework regulates the frequency response of ND filters for arbitrary derivative-order and cutoff frequency selected parameters relying on interpolating power polynomials and maximally flat design techniques. Compared with the state-of-the-art solutions, such as Gaussian kernels, the proposed ND filter is sharply localized in the Fourier domain with ripple-free artifacts. Here, we construct 2D MaxFlat kernels for image directional differentiation to calculate image differentials for arbitrary derivative order, cutoff level and steering angle. The resulted kernel library renders a new solution capable of delivering discrete approximation of gradients, Hessian, and higher-order tensors in numerous applications. We tested the utility of this library on three different imaging applications with main focus on the unsharp masking. The reported results highlight the high efficiency of the 2D MaxFlat kernel and its versatility with respect to robustness and parameter control accuracy. [ABSTRACT FROM PUBLISHER]

Societal freedoms need to be understood both in terms of structural pluralism and directional pluralism. The former concerns personal freedom as well as the plurality of societal entities present in a differentiated society while the latter accounts for the freedom allowing diverging life- and worldviews to live to the full their deepest convictions in all walks of life, liberated from any form of unlawful interference. Acknowledging this distinction exceeds the limitations of the underlying atomistic (individual-centric) underpinnings of modem social contract theories (such as is found in Locke's view: "for all being kings as much as he, every man his equal"). This spirit is breathed by the so-called "conscience clause" almost hundred years ago introduced in the Private Law of South African universities (it started with the University of South Africa and the University of Stellenbosch, respectively in 1916 and 1917). The University of Potchefstroom as well as the Homeland universities did not inherit this clause. It is striking that universities, as academic institutions, were confronted with a religious clause, as if a university is a faith community. The conscience clause is still at home within the positivist idea of an un-prejudiced, objective and neutral science mediated by factual sense data. This positivist view was thoroughly criticised by the philosophy of science of the previous century (Kuhn, Popper, Lakatos, Toulmin, Stegmüller and many others). The idea of an autonomous individual gave way to the collective identity of supra-individual institutions, which had to account for their directional orientation by formulating mission and vision statements. It falls outside the sphere of competence of the government to interfere with the directional choice of a citizen in non-political contexts -- and the same applies to the domain of the public opinion. In the Netherlands it is still possible, for example, to promote the ideals of a specific trend of thought in scholarly pursuits by establishing special chairs at public universities. During the third decade of the 20,h century this was done by the Bolland Foundation which established in Leiden a special chair in Hegelian Philosophy. At the moment there are still special chairs in Reformational Philosophy at six public universities in the Netherlands. Academic freedom, individually and collectively, entails directional freedom. Currently the Darwinian orthodoxy in the USA controls the teaching of biology to such an extent that no biology teacher is even allowed to raisescientific criticism against Darwinism. It contains a threat to various freedoms, amongst which are treedom oi speech, freedom of thought, academic freedom, and religious freedom. This is amply demonstrated by Jerry Bergman in his work, Slaughter o f the Dissidents (2008). Similar to what was required in the Soviet Union of the 20thcentury, biology teachers in the USA now have to be atheists. The "in-between-position" of theistic evolutionists, which holds that God directed evolution (a process which is actually, according to Darwinism, undirected) is even worse in the eyes of Darwinian orthodoxy, because it violates the church-state division by smuggling God back into the classroom. In his Review of Bergman's book, Brian Thomas remarks that all "Darwin Doubters" are seen as "creationists." Althouh highly qualified, these scientists were threatened, while losing opportunities, jobs and even whole careers because of their doubts about the standard evolutionary story. Bergman remarks that this situation already "claimed many thousands of victims." This completes the circle, reminding us of what happened in early Christendom during the persecution of Christians. Fortunately, there is positive legislation in South Africa which avoids this extreme position. However, a recent experience of the author of this article apparently highlights the emergence of something similar to what is happening in the USA. [ABSTRACT FROM AUTHOR]

Interleukin-12 (IL-12), a member of interleukin family, plays a critical role in immune responses and anti-tumor activity. In this study, the effects of IL-12 on monocytic tumor cell lines differentiation to macrophagocyte and its likely mechanism was investigated. We examined the differentiation markers, morphological and functional changes, and possible mechanism in IL-12-treated THP-1 and U937 cells. It was found that IL-12 could up-regulated macrophage surface marker CD68 and CD11b expression in a time-dependent manner. Morphologically, after IL-12 treatment, THP-1 and U937 cells became round or irregular shape, even stretched many cell membrane protuberances; some cell nuclei became fuzzy or completely disappeared, and the chromatin appeared dense and cordlike. Furthermore, IL-12-induced monocytic tumor cell differentiation was accompanied by the growth arrest with G1-phase accumulation and S-phase reduction; apoptosis increased with anti-apoptosis protein Bcl-2 down-expression and pro-apoptosis protein Fas up-regulation, and enhanced phagocytosis function. The IL-12-induced macrophage differentiation of THP-1 and U937 cells was associated with the up-regulation of c-fms expression and the CSF-1R Tyr 809 site phosphorylation. These findings have revealed that IL-12 could induce monocytic tumor cells directional differentiation into macrophage-like cells, and its mechanism is possible connected with the up-regulation of c-fms expression and the phosphorylation of CSF-1R Tyr-809 site. [ABSTRACT FROM AUTHOR]

The purpose of this study was to investigate the repair of the osteoarthritis(OA)-induced cartilage injury by transfecting the new TGF-β3 fusion protein (LAP-MMP-mTGF-β3) with targeted therapy function into the bone marrow-derived mesenchymal stem cells (MSCs) in rats. The recombinant of pIRES-EGFP-MMP was constructed by combination of DNA encoding MMP enzyme cutting site and eukaryotic expression vector pIRES-EGFP. LAP and mTGF-β3 fragments were obtained from rat embryos by RT-PCR and inserted into the upstream and downstream of MMP from pIRES-EGFP-MMP respectively, so as to construct the recombinant plasmid of pIRES-EGFP-LAP-MMP-mTGF-β3. pIRES-EGFP-LAP-MMP-mTGF-β3 was transfected into rat MSCs. The genetically modified MSCs were cultured in medium with MMP-1 or not. The transfected MSCs were transplanted in the rat OA models. The OA animal models were surgically induced by anterior cruciate ligament transaction (ACLT). The pathological changes were observed under a microscope by HE staining, Alcian blue, Safranin-fast Green and graded by Mankin's scale. pIRES-EGFP-LAP-MMP-mTGF-β3 was successfully constructed by means of enzyme cutting and sequencing, and the mTGF-β3 fusion protein (39 kD) was certified by Western blotting. Those genetically modified MSCs could differentiate into chondrocytes induced by MMP and secrete the relevant-matrix. The transfected MSCs could promote chondrogenesis and matrix production in rat OA models in vivo. It was concluded that a new fusion protein LAP-MMP-mTGF-β3 was constructed successfully by gene engineering, and could be used to repair the OA-induced cartilage injury. [ABSTRACT FROM AUTHOR]

To remove the distortion caused by relative moving during taking pictures, assuming that the original image obeys isotropic Markov process with rank one, an efficient identification approach is proposed for motion-blurred direction from motion-blurred image with a 3 × 3 directional differentiation matrix and the bilinear interpolation. A proper choice of differentiation and the method of "first identifying cursorily and then carrying out the refined identification" can help to identify the direction of the motion, and then the blurred image can be rotated to a horizontal axis and the image restoration can be made out easily in one dimension. This optimized identification of motion blur direction has high precision and stability compared with Yitzhaky's method. A correlation function between the pixels in the blur extent of a blurred image is applied to figure out the blur extent. With these two parameters (the motion blur direction and the blur extent), the PSF (point spread function) can be obtained and the restoration by filters can be carried out with good results. [ABSTRACT FROM AUTHOR]

Abstract: The aim of this study was to investigate the possible inducible effects and to clarify the modulation by icariin of cell cycle and p53 expression in the differentiation of embryonic stem cells into cardiomyocytes in vitro. Embryonic stem cells were cultivated as embryoid bodies in hanging drops and induced to differentiate into cardiomyocytes by icariin at 10−7 M. Cardiomyocytes were characterized by the expression of sarcomeric proteins, α-actinin and cardiac troponin T, by immunocytochemistry. Flow cytometry revealed that 10−7 M icariin treatment for 48 h significantly induced the accumulation of cells in G0/G1 and reduced the proportion of cells in S phase. A marked increase in apoptosis rate was observed 48 h after icariin treatment. Icariin resulted in significantly increased expressions of p53 mRNA and protein, as determined by reverse transcription-polymerase chain reaction and Western blot analysis. During day 7+0 and 7+9 cardiac developmental stage, 10−7 M icariin increased the level of p53 mRNA, but caused a parallel decrease in the level of p53 protein. In conclusion, icariin at 10−7 M facilitated the directional differentiation of embryonic stem cells into cardiomyocytes. Results showed p53 to be an important regulator in the differentiation in embryonic stem cells treated with 10−7 M icariin, controlling or adjusting the balance between differentiated cells and cells undergoing apoptosis. [Copyright &y& Elsevier]

Aim: To investigate the possible inducible effects of icariin, icaritin, and desmethylicaritin on the directional differentiation of embryonic stem (ES) cells into cardiomyocytesin vitro.Methods: ES cells were cultivated as embryoid bodies (EBs) in hanging drops with icariin, icaritin, or desmethylicaritin. ES cells treated with retinoic acid and with solvent were used as positive and negative controls, respectively. The cardiomyocytes derived from the ES cells were verified using immunocytochemistry. The expression of cardiac developmentaldependent genes was detected using the reverse transcription-polymerase chain reaction (RT-PCR) method. Cell cycle distribution and apoptosis were analyzed using flow cytometry to determine the partly inducible effect mechanisms involved.Results: The total percentage of beating EBs treated with 1×10−7 mol/L icariin, icaritin, or desmethylicaritin was 87% (P<0.01), 59% (P<0.01), and 49%, respectively. All the beating cardiomyocytes derived from the ES cells expressed cardiac-specific proteins forα-actinin and troponin T. Among them, 1×10−7 mol/L icariin treatment resulted in a significantly advanced and increased mRNA level ofα-cardiac major histocompatibility complex (MHC) and myosin light chain 2v (MLC-2v) in EBs in the early cardiac developmental stage. Before shifting to the cardiomyocyte phenotype, icariin could evoke the accumulation of ES cells in G0/G1 and accelerate apoptosis of the cell population (P<0.05).Conclusion: Icariin facilitated the directional differentiation of ES cells into cardiomyocytes at a concentration of 1×10−7 mol/L. The promoting effect of icariin on cardiac differentiation was related to increasing and accelerating gene expression ofα-cardiac MHC and MLC-2v, as well as regulating the cell cycles and inducing apoptosis. [ABSTRACT FROM AUTHOR]

Idiopathic pulmonary fibrosis (IPF) is a type of pulmonary disease that progresses acutely or slowly into irreversible pulmonary diseases, resulting in the end severe damages to patients' lung functions, as well as deaths. At present, the pathogenesis of pulmonary fibrosis is still not clear and there is no effective therapeutic measure available to control the progression of the disease. Research findings indicate that stem cells, being the origin of all cells of organisms, participate in the development of individuals at various stages and play an important role in repairing pulmonary tissue damage. Stem cells are attracting growing attention in the field of regenerative medicine, providing new ideas for treating IPF with transplanted stem cells. Herein, in order to better explore the potential applications of stem cell transplantation in treating IPF, we attempt to summarize preliminary studies of stem cell-mediated pulmonary remodeling after IPF, as well as cutting-edge clinical trials in stem cell-based IPF therapy., (Copyright© by Editorial Board of Journal of Sichuan University (Medical Sciences).)

Interleukin 17 (IL-17) and its main producer, T cell receptor γδ cells, have neurotoxic effects in the pathogenesis of intracerebral hemorrhage (ICH), aggravating brain injuries. To investigate the correlation between IL-17 and ICH, we dynamically screened serum IL-17 concentrations using enzyme-linked immunosorbent assay and explored the clinical values of IL-17 in ICH patients. There was a significant negative correlation between serum IL-17 level and neurological recovery status in ICH patients (r = -0.498, P < 0.01). To study the neurotoxic role of IL-17, C57BL/6 mice were used to establish an ICH model by injecting autologous blood into the caudate nucleus. Subsequently, the mice were treated with mouse neural stem cells (NSCs) and/or IL-17 neutralizing antibody for 72 hours. Flow cytometry, brain water content detection, Nissl staining, and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling results indicated that NSC transplantation significantly reduced IL-17 expression in peri-hematoma tissue, but there was no difference in T cell receptor γδ cells. Compared with the ICH group, there were fewer apoptotic bodies and more Nissl bodies in the ICH + NSC group and the ICH + NSC + IL-17 group. To investigate the potential effect of IL-17 on directional differentiation of NSCs, we cultured mouse NSCs (NE-4C) alone or co-cultured them with T cell receptor γδ cells, which were isolated from mouse peripheral blood mononuclear cells, for 7 days. The results of western blot assays revealed that IL-17 secreted by T cell receptor γδ cells reduced the differentiation of NSCs into astrocytes and neurons, while IL-17 neutralization relieved the inhibition of directional differentiation into astrocytes rather than neurons. In conclusion, serum IL-17 levels were elevated in the early stage of ICH and were negatively correlated with outcome in ICH patients. Animal experiments and cytological investigations therefore demonstrated that IL-17 probably has neurotoxic roles in ICH because of its inhibitory effects on the directional differentiation of NSCs. The application of IL-17 neutralizing antibody may promote the directional differentiation of NSCs into astrocytes. This study was approved by the Clinical Research Ethics Committee of Anhui Medical University of China (For human study: Approval No. 20170135) in December 2016. All animal handling and experimentation were reviewed and approved by the Institutional Animal Care and Use Committee of Anhui Medical University (approval No. 20180248) in December 2017., Competing Interests: None

Objective: To investigate the ability of gene-loaded lipopolysaccharide-amine nanopolymersomes (LNPs) in inducing osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) by in vitro gene transfection, where LNPs were used as a non-viral cationic carrier, and their properties were optimized during synthesis., Methods: LNPs were synthesized by a graft-copolymerization method, and the effects of different pH environments during synthesis on physicochemical properties of LNPs and LNPs/plasmid of bone morphogenetic protein 2-green fluorescent protein (pBMP-2-GFP) complexes were explored. Then, optimized LNPs with maximum transfection efficiency and safe cytotoxicity in rat BMSCs were identified by cytotoxicity and transfection experiments in vitro . Thereafter, the optimized LNPs were used to mediate pBMP-2-GFP to transfect rat BMSCs, and the influences of LNPs/pBMP-2-GFP on osteogenic differentiation of BMSCs were evaluated by monitoring the cell morphology, concentration of BMP-2 protein, activity of alkaline phosphatase (ALP), and the formation of calcium nodules., Results: The nitrogen content, particle size, and zeta potential of LNPs synthesized at pH 8.5 were lower than those of the other pH groups, with the lowest cytotoxicity (96.5%±1.4%) and the highest transfection efficiency (98.8%±0.1%). After transfection treatment, within the first 4 days, BMSCs treated by LNPs/pBMP-2-GFP expressed BMP-2 protein significantly higher than that treated by Lipofectamine2000 (Lipo)/pBMP-2-GFP, polyethylenimine 25K/pBMP-2-GFP, and the blank (non-treated). At 14 days after transfection, ALP activity in BMSCs treated by LNPs/pBMP-2-GFP was higher than that treated by Lipo/pBMP-2-GFP and the blank, comparable to that induced by osteogenic medium; with alizarin red staining, visible calcium nodules were found in BMSCs treated by LNPs/pBMP-2-GFP or osteogenic medium, but absent in BMSCs treated by Lipo/pBMP-2-GFP or the blank with apoptosis. At 21 days after transfection, transparent massive nodules were discovered in BMSCs treated by LNPs/pBMP-2-GFP, and BMSCs exhibited the morphologic features of osteoblasts., Conclusion: LNPs synthesized at pH 8.5 has optimal transfection efficiency and cytotoxicity, they can efficiently mediate pBMP-2-GFP to transfect BMSCs, and successfully induce their directional osteogenic differentiation, whose inducing effect is comparable to the osteogenic medium. The results suggest that gene transfection mediated by LNPs may be a convenient and effective strategy in inducing directional differentiation of stem cells.