Further suggesting an involvement of astrocytic GJC in epilepsy, is the fact that the expression of astrocytic Cx mRNAs (Cxs 30 and 43) is several fold higher than that of neuronal Cx mRNAs (Cxs 36 and 45), and the number of glial cells outnumber neuronal cells in mammalian hippocampal and cortical tissue. Pannexin expression is also increased in both animal and human epileptic tissues. Specific Cx43 mimetic peptides, Gap 27 and SLS, inhibit the docking of astrocytic connexin Cx43 proteins from forming intercellular gap junctions (GJs), diminishing spontaneous seizures. Besides GJs, Cx membrane hemichannels in glia and Panx membrane channels in neurons and glia are also inhibited by traditional gap junctional pharmacological blockers. Although there is no doubt that connexin-based GJs and hemichannels, and pannexin-based membrane channels are related to epilepsy, the specific details of how they are involved and how we can modulate their function for therapeutic purposes remain to be elucidated.Enhanced gap junctional communication (GJC) between neurons is considered a major factor underlying the neuronal synchrony driving seizure activity. In addition, the hippocampal sharp wave ripple complexes, associated with learning and seizures, are diminished by GJC blocking agents. Although gap junctional blocking drugs inhibit experimental seizures, they all have other non-specific actions. Besides interneuronal GJC between dendrites, inter-axonal and inter-glial GJC is also considered important for seizure generation. Interestingly, in most studies of cerebral tissue from animal seizure models and from human patients with epilepsy, there is up-regulation of glial, but not neuronal gap junctional mRNA and protein. Significant changes in the expression and post-translational modification of the astrocytic connexin Cx43, and Panx1 were observed in an  When one thinks of the “connections” between epilepsy and gap junctions (GJs), the usual interpretation is that the GJs form direct intercellular cytoplasmic connections between neurons, promoting the hypersynchronous neuronal activity associated with seizures. However, as this review discusses, that concept is rather naïve and greatly complicated by much new data concerning the potential roles of gap junctional communication (GJC) and membrane Cx hemichannels and pannexin channels. , a K+ channel blocker which induces seizures, have shown that dephosphorylation of connexin 43 associated with astrocytic swelling, resulted in reduction of astrocytic gap junction permeability  Figures ,C is higin vivo, their occurrence was completely inhibited by pharmacological agents known to block GJs, such as octanol, carbenoxolone, halothane, and low pH.The mechanisms responsible for ripple generation are not fully understood, however a prominent theory suggests that gap junction proteins play a critical role Figures ,C. Traubin vivo ripple oscillations were unaltered in terms of power, frequency, or probability of occurrence  and lanthanum (La3+) block hemichannels without inhibiting gap junction or Panx1 channels, these ions are not specific to GJ proteins since they also block other channels such as Ca2+ channels (Mlinar and Enyeart, Several of the compounds from the fenemate family of blockers block Panx1 hemichannels in the following order: mefloquine > carbenoxolone > flufenamic acid (Iglesias et al., 2+ or low Ca2+ seizure models, or the reduced extracellular Ca2+, which is seen during seizures in several seizure models (Lux and Heinemann, Connexin mimetic peptides, which correspond to the extracellular loops of connexins, are a class of specific and reversible inhibitors of GJC (Evans and Boitano, Developmental age plays a role in gap junctional expression and seizure susceptibility. It is well-known that immature humans and animal models are more susceptible to undergoing seizures than mature adults (Rakhade and Jensen, in vitro Co2+-induced seizures (Mylvaganam et al., Analysis of the mRNA expressions of different Cxs and Panxs in the hippocampal tissues from 15 day mice, showed the highest expression of astrocytic Cxs 43 and 30, followed by Panxs, and the lowest expression was observed in the neuronal Cxs 36, 40, and 45, a few orders of magnitude lower than the pannexins and astrocytic connexons (Mylvaganam et al., Studies have shown changes in the expression of GJ mRNAs and proteins or changes in the post-translational modification form of the Cx43 and Panx protein in epileptic animal models Table . Seizurein vivo, since they showed pilocarpine-induced seizures in mice were enhanced following administration of P2X7R antagonists or by gene silencing of P2X7R or Panx1 in WT in a process mediated by PKC via intracellular Ca2+ release (Kim and Kang, Among the many Cxs present in the brain, studies have reported alterations only in Cx30, 32, 36, and 43 Table . InteresGap junction expression studies in human epileptic tissue have demonstrated no change (Elisevich et al., Whether these changes in gene expressions have any direct effect on seizure generation is unclear. These changes might be pro-epileptic responses contributing to the pathogenesis of the disease, or adaptive responses to cope with the pathologic condition. Confounding the interpretation of human data is the fact that epilepsy is not a single condition, but a large group of highly heterogeneous disorders, which have in common an abnormally increased predisposition to seizures (Fisher et al., At present, there are at least 10 distinct diseases known to be associated with gene mutations in connexins, some of which are associated with epilepsy. Connexin-linked diseases caused by gene mutations or altered connexin expression, protein assembly or localization, will ultimately impact the rest of the proteome, which can influence the manifestation of the pathology Laird, . In humaTSC1 and TSC2 are found in tuberous sclerosis, and epilepsy is one of its major manifestations. Deleting TSC1 in astrocytes caused seizures attributed to diminished astrocytic GJ intercellular communication and impaired potassium buffering (Xu et al., Mutations of the tumor suppressor genes In brief, the roles of GJs, connexins, and pannexins in epilepsy remain unclear. The lack of specificity of pharmacological agents now used to block or enhance GJC is a challenge if one wants to understand the role of a particular type of connexin-based GJ, and if wants to develop a specific therapeutic tool. The fact that most of these agents affect the Cx hemichannels and the Panx membrane channels further confuses the interpretation of experimental data. The ongoing development of peptides targeting specific Cxs and Panxs will permit a more precise dissection of the functions of the different Cx and Panx species. Although the gap junctional blocking drugs are almost always anticonvulsant, there is not as of yet an anticonvulsant drug on the market that is reputed to have gap junctional blocking properties, although both acetazolamide and topiramate inhibit carbonic anhydrase activity which should cause an intracellular acidosis thereby blocking GJC. Another problem is the overwhelming evidence that tissue from animal models and human epileptics show increases in Cxs expression in glia but not in neurons. This begs the question as to what are the roles of glia in seizure generation. If applying a gap junctional blocking agent to epileptic tissue is blocking interglial GJC, then what is the underlying anticonvulsant mechanism? Or could these agents have their major anticonvulsant action by blocking the conductance of Cx hemichannels and/or Panx channels? The scientific community is just now starting to consider the possible role of these membrane channels in the pathophysiology of epilepsy. In summary, although there is no doubt that GJs, connexins and pannexins are intimately related to epilepsy and seizure generation, the specific details of exactly how they are involved and how we can modulate their function for therapeutic purposes remain to be elucidated.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The mammalian cerebral cortex contains a high variety of neuronal subtypes that acquire precise spatial locations and form long or short-range connections to establish functional neuronal circuits. During embryonic development, cortical projection neurons are generated in the areas lining the lateral ventricles and they subsequently undergo radial migration to reach the position of their final maturation within the cortical plate. The control of the neuroblast migratory behavior and the coordination of the migration process with other neurogenic events such as cell cycle exit, differentiation and final maturation are crucial to normal brain development. Among the key regulators of cortical neuron migration, the small GTP binding proteins of the Rho family and the atypical Rnd members play important roles in integrating intracellular signaling pathways into changes in cytoskeletal dynamics and motility behavior. Here we review the role of Rnd proteins during cortical neuronal migration and we discuss both the upstream mechanisms that regulate Rnd protein activity and the downstream molecular pathways that mediate Rnd effects on cell cytoskeleton. During the development of the central nervous system, neural progenitors undergo a sequence of distinct cellular events to give rise to the vast array of neurons that populate the entire brain. In the cerebral cortex, excitatory projection neurons, which constitute the majority of cortical neurons, are generated from neural stem/progenitor cells located in the areas lining the lateral ventricles, the ventricular (VZ) and subventricular zones (SVZ) of the dorsal telencephalon , these cells self-renew to expand the progenitor pool and then convert into cells with glia-like features, the radial glial cells. A typical feature of these two populations of progenitors is the apico-basal movement of their nuclei in coordination with cell cycle progression, a phenomenon known as interkinetic nuclear migration (INM)  networks involved in this process  GTPases belong to the large superfamily of small GTP binding proteins, whose founding member is Ras  and Ascl1 directly bind to E-box DNA sequences within enhancers located 3′ to the Rnd2 and Rnd3 gene, respectively  abolish its plasma membrane association and impairs its ability to rescue the migratory activity of Rnd3-silenced neurons, thus demonstrating that membrane association is required for Rnd3 activity in migrating neurons  that is preferentially associated to the plasma membrane is more efficient than wild-type Rnd3 in rescuing the cortical migration defects induced by Rnd3 silencing  and they have been shown to influence Rnd3 localization at the plasma membrane. Rnd3 can be phosphorylated by ROCKI or PKCα on multiple sites - LIMK (Lim Kinase)—cofilin pathway , which constitutively depolymerizes actin, together with Rnd3 shRNA completely rescues the migration defects induced by Rnd3 silencing. This suggests that when Rnd3 is silenced, the RhoA-cofilin-mediated excessive polymerization of actin molecules hampers the motility behavior of migrating neurons. Interestingly, co-electroporation of the cofilin mutant with Rnd3 shRNA also rescues the defects that Rnd3 silencing produces during INM in VZ progenitor cells  in migrating neurons , which cannot bind to p190RhoGAP, is as active as wild type Rnd2 in rescuing the neuronal migration defects induced by Rnd2 silencing in the cortex. Therefore, even if Rnd2 can interact with p190RhoGAP, this interaction does not mediate Rnd2 function in this context. It is possible that Rnd2 works via interaction with a different RhoGAP. One candidate is MgcRacGAP, which has been found associated to Rnd2 in male germ cells  that cannot bind to p190RhoGAP in rescue experiments failed to replace Rnd3 function in migrating neurons  (Wennerberg et al., In contrast to Rnd2, Rnd3 requires the interaction with p190RhoGAP for its pro-migratory activity in the cortex. Indeed, an Rnd3 mutant form (Rnd3Over the past few years, Rnd proteins have been shown to constitute important functional components of the plexin-semaphorin signaling pathways Chardin, . PlexinsPlexins contain a binding site for Rho GTPases in the middle of their intracellular domain through which they recruit several Rho GTPases, including Rnd proteins. Several evidences indicate that preferential interactions occur between certain members of the Plexin and the Rnd families Table . For exaRnd1 binding to PlexinB1 has been shown to open the conformation of the receptor and to allow the transmission of the downstream signaling. This synergistic interaction is essential to drive cell contraction in COS cells and to induce growth cone collapse during axon guidance Chardin, . In migrRnd2 has also been found associated with Plexin members like PlexinB1 and PlexinD1. However, in contrast to Rnd3, which co-localizes with PlexinB2 at the plasma membrane in primary cortical neurons, Rnd2 is not found at the plasma membrane (Pacary et al., Altogether, these studies suggest that Rnd2 and Rnd3 promote cortical neuron migration by distinct mechanisms that may involve selective interactions with different members of the plexin family of transmembrane receptors in different subcellular compartments.in utero electroporation of the murine embryonic cerebral cortex, have greatly advanced our understanding of the molecular pathways operating in migrating neurons (Loturco et al., In the last decade, the introduction and constant refinement of new technologies, such as Rnd2 and Rnd3 expression in the cortex is under the transcriptional control of the proneural factors Neurog2 and Ascl1, respectively. These factors are well known master regulators of neuronal differentiation and activate a transcriptional program of neurogenesis in neural progenitors (Bertrand et al., At the molecular level, Rnd2 and Rnd3 control distinct steps of radial migration, by interfering with RhoA activity in different subcellular compartments. The bHLH transcriptional factors Ascl1 and Neurog2 induce the expression of Rnd proteins as a strategy to repress RhoA during radial migration (Hand et al., These studies performed in neuronal cells will contribute to a better understanding of the regulatory function of Rnd proteins in the migration of other cell types. Indeed, Rnd proteins, especially Rnd3, have been shown to also control the migration of non-neuronal cell types, such as epithelial cells (Guasch et al., In conclusion, it is becoming evident that Rnd proteins play important roles in cell migration during mammalian cortical development and in particular, considering their relatively recent evolution, it is possible that they might be involved in mechanisms of brain developmental and neuronal plasticity that are exclusive to vertebrate organisms.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


During hypovolemic shock, skeletal muscle blood flow could be redistributed to vital organs via vasoconstriction in part evoked by activation of the innervating sympathetic nerve activity. However, it is not well known whether this mechanism operates during anaphylactic shock. We determined the femoral artery blood flow (FBF) and lumbar sympathetic nerve activity (LSNA) mainly regulating the hindquater muscle blood flow during anaphylactic hypotension in anesthetized rats. Anesthetized Sprague-Dawley rats were randomly allocated to the following groups (n = 7/group): (1) non-sensitized, (2) anaphylaxis, (3) anaphylaxis-lumbar sympathectomy (LS) and (4) anaphylaxis-sinoaortic denervation (SAD) groups. Anaphylaxis was induced by an intravenous injection of the ovalbumin antigen to the sensitized rats. The systemic arterial pressure (SAP), heart rate (HR), central venous pressure (CVP), FBF and LSNA were continuously measured. In the anaphylaxis group, LSNA and HR increased, while SAP and FBF decreased after antigen injection. In the anaphylaxis-SAD group, LSNA did not significantly change during the early phase, but the responses of SAP and FBF were similar to those in the anaphylaxis group. In the anaphylaxis-LS group, both FBF and SAP decreased similarly to the anaphylaxis group during anaphylactic hypotension. These results indicated that LSNA increased via baroreceptor reflex, but this sympathoexcitation or LS did not affect antigen-induced decreases in FBF or SAP. Lumbar sympathetic nerves are not involved in regulation of the blood flow to the hindlimb or systemic blood pressure during anaphylactic hypotension in anesthetized rats. Anaphylactic shock is potentially life threatening, but the pathophysiology has not been fully clarified. During hemorrhagic shock, cardiac output is preferentially directed toward vital organs. Myocardial and cerebral perfusions are indeed maintained, whereas perfusion of liver, intestines, kidneys, and muscles is significantly reduced due to vasoconstriction , 2. ActuMale Sprague-Dawley rats, weighing 300-400g, were housed in a room maintained at 24±1°C and illuminated for 12 hours every day. Food and water were freely available. All the animal care and handling procedures were approved by the animal research committee of Kanazawa Medical University.Rats in the anaphylaxis, lumbar sympathectomy (LS) and sinoaortic denervation (SAD) groups were sensitized though subcutaneous injection of an emulsion made by mixing equal volumes of complete Freund’s adjuvant (0.5ml) with 1 mg ovalbumin  dissolved in physiologic saline (0.5 ml) two weeks before experiment .Rats were anesthetized with an intraperitoneal injection of pentobarbital sodium (50 mg/kg). The adequacy of anesthesia was monitored by the stability of blood pressure and respiration under control conditions and during a pinch of the hind paw. When a significant change of blood pressure (+5 mmHg) was detected by a paw pinch, additional pentobarbital sodium (5 mg/kg) was intraperitoneally injected. The trachea was intubated to facilitate spontaneous breathing. Polyethylene catheters were inserted into the left femoral vein, right jugular vein and left carotid artery for intravenous injections, central venous pressure (CVP) measurement and systemic arterial pressure (SAP) measurement, respectively. A pulsed Doppler flow probe  was placed on the right femoral artery for continuous measurement of the femoral arterial blood flow (FBF). The animal was placed on a heating pad to maintain the body temperature at 36–37°C. The SAP and CVP were continuously measured with pressure transducers , and the reference level was set at the level of the right atrium. Heart rate (HR) was measured by triggering SAP. These hemodynamic variables along with FBF were digitally displayed and recorded by Power Lab . The femoral arterial vascular resistance (FVR) was calculated by the following equation: FVR = (SAP − CVP)/FBF.LSNA was measured as follows: Following an abdominal mid-line incision at supine position, the intestines were retracted, and the abdominal aorta and vena cava caudal to the renal vessels were pulled aside to expose a lumbar nerve. The left lumbar sympathetic trunk from L3 to L4 supplying skeletal muscle , 11 was All the rats were allowed to stabilize for 10–30 min after surgery and then sodium nitroprusside  was injected intravenously to confirm baroreceptor-mediated sympathoexcitation. The occurrences of baroreceptor reflex in the non-sensitized group and anaphylaxis group were confirmed by increased LSNA and HR . Next, tIn the anaphylaxis-LS group, denervation was accomplished by cutting bilateral lumbar sympathetic nerves from L3 to L4 along the abdominal aorta and vena cava. Immediately the abdominal aorta and vena cava was painted with 10% phenol diluted with ethanol. Careful application of phenol was taken to protect the adjacent tissue from the harmful effects of phenol. The occurrence of baroreceptor reflex in the LS group was confirmed by reflex tachycardia.In the anaphylaxis-SAD group, after the neurovascular trunk in the neck was exposed, the carotid sinus, aortic depressor and recurrent laryngeal nerves were sectioned bilaterally to complete the baroreceptor denervation. Immediately the bilateral carotid arteries were carefully painted with 10% phenol diluted with ethanol to avoid physical and chemical damage to the vagal nerves. The rats were allowed to recover for 30–60 min after denervation surgery. Successful baroreceptor denervation was confirmed by the absence of reflex increases in HR and LSNA in response to the hypotension induced by SNP .P < 0.05.All results are expressed as the means ± SE. Comparison of individual values within a group was made by the repeated-measures ANOVA followed by the Fisher post hoc test. Comparison of sympathetic and hemodynamic responses to the antigen between the four groups was made by two way ANOVA with Bonferroni post hoc test. Differences were considered statistically significant at Examples of the response of LSNA, HR, SAP, FBF and CVP to an injection of antigen into a non-sensitized rat and an ovalbumin-sensitized rat were shown in In the anaphylaxis-SAD group, LSNA showed no significant changes in response to an antigen injection despite the occurrence of SAP fall during the early phase (0–20 min after antigen injection). There were significant differences in LSNA during this period between the SAD and anaphylaxis groups Figs  and 2A. In order to determine the role of LSNA in the responses of FBF , FVR Fi and SAP In the present study, we investigated lumbar sympathetic and femoral vascular responses to anaphylactic hypotension in the ovalbumin-sensitized anesthetized rats, and obtained the following findings: 1) After the antigen injection, LSNA was increased and FBF was substantially decreased, while FVR initially decreased and then returned to the baseline. 2) SAD inhibited the antigen-induced activation of LSNA only at the early phase. 3) Neither SAD nor LS affected the responses of FVR, FBF or SAP to anaphylaxis. These findings indicate that LSNA does not participate in regulation of the blood flow to hindquarter skeletal muscles during anaphylactic hypotension in anesthetized rats. To the best our knowledge, this is the first experimental study to clarify, by measuring simultaneously the femoral arterial blood flow and LSNA, the relationship between the blood flow to hindlimb or hindquarter skeletal muscles and their relevant sympathetic outflow during anaphylactic hypotension.Previously, it was reported that rat anaphylactic shock activated the sympathetic nervous system, as evidenced by the elevated plasma concentrations of epinephrine and norepinephrine , 12 and In contrast to the dependency of the sympathoexcitation of LSNA via the baroreceptor reflex, the anaphylaxis-induced increase in renal sympathetic nerve activity was not affected by SAD in our previous study . In addi2 [4 [Generally, during severe hypotension, as observed in all forms of systemic shock, the blood flow to peripheral organs such as skeletal muscle is reduced to keep constant blood flow supplied to vital organs such as brain and heart. In fact, skeletal muscle blood flow in anesthetized rats was decreased under the anaphylactic shock . To redi2  and leuk2 [4  were als2 [4 , 22. 2) 2 [4 . Actuall2 [4 , 25. 3) 2 [4 . TherefoAn interesting finding of the present study is that the femoral arterial vasodilatation, as evidenced by increased FBF and decreased FVR, occurred immediately after the antigen injection, and simultaneously with the initial fall of SAP. The similar vasodilatation is observed in the arteries of other vascular beds, such as the hepatic artery and mesenteric artery during anaphylaxis in anaesthetized rats , 27. ThiThe blood flow to the hindlimb was decreased by 63.9% in the present study. This reduction of FBF was much smaller than that of the splanchnic vascular beds as observed in our previous studies, in which the same rat anaphylaxis models were used: the blood flow reductions of the mesenteric artery  and the In summary, the present study shows that anaphylactic hypotension in anesthetized rats is accompanied by FBF reduction and lumbar sympathoexcitation which depends on the baroreceptor reflex only in the early phase of anaphylaxis. However, the FBF and SAP responses were not mediated by either the lumbar sympathetic nerves or the baroreceptor reflex. Thus, we conclude that lumbar sympathetic nerves are not critical to regulation of the blood flow to the hindlimb or blood pressure during anaphylactic hypotension in anesthetized rats.


In silico analysis is essential to investigate putative genetic and epigenetic elements of tumor suppressor genes such as Rb1 gene. This may contribute genetic and epigenetic information modulating tissue-specific transcripts and expression levels of genes . In order to gain substantial coverage of the human epigenome, the IHEC is planning to decipher at least 1000 epigenomes. These multilayer-omics analyses including genome, epigenome, transcriptome, proteome and metabolome are important for elucidating the molecular carcinogenesis and for exploring biomarkers and therapeutic targets for human cancers  was established to coordinate the production of reference maps of human epigenomes for key cellular states . CSCs express specific cell surface markers including CD44, CD133, and EpCAM. Recently, a novel 3D culture method for stem cells called “organoid culture” has been developed. This culture method uses a serum-free medium that includes only identified growth factors such as R-spondin 1, EGF, and Noggin. R-spondin 1 is a ligand for Lgr5, which is a marker for intestinal stem cells and an essential factor to activate Wnt signal in intestinal crypts. Intestinal organoid culture enabled to expand normal or tumor epithelial cells let-7 and miR-34a have been implicated in the regulation of CSC properties by suppression of their target genes such as HMGA2, RAS, NOTCH1, and CD44. The modulation of CSC gene expression by miRNAs could be a novel therapeutic strategy targeting CSCs  such as MEG3 and CRNDE are also dysregulated in glioma tissues and may be associated with the stemness of glioma cells (Katsushima and Kondo, miRNAs including The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Possessing the ability of self-renewal with immortalization and potential for differentiation into different cell types, stem cells, particularly embryonic stem cells (ESC), have attracted significant attention since their discovery. As ESC research has played an essential role in developing our understanding of the mechanisms underlying reproduction, development, and cell (de)differentiation, significant efforts have been made in the biomedical study of ESC in recent decades. However, such studies of ESC have been hampered by the ethical issues and technological challenges surrounding them, therefore dramatically inhibiting the potential applications of ESC in basic biomedical studies and clinical medicine. Induced pluripotent stem cells (iPSCs), generated from the reprogrammed somatic cells, share similar characteristics including but not limited to the morphology and growth of ESC, self-renewal, and potential differentiation into various cell types. The discovery of the iPSC, unhindered by the aforementioned limitations of ESC, introduces a viable alternative to ESC. More importantly, the applications of iPSC in the development of disease models such as neurodegenerative disorders greatly enhance our understanding of the pathogenesis of such diseases and also facilitate the development of clinical therapeutic strategies using iPSC generated from patient somatic cells to avoid an immune rejection. In this review, we highlight the advances in iPSCs generation methods as well as the mechanisms behind their reprogramming. We also discuss future perspectives for the development of iPSC generation methods with higher efficiency and safety. Due to their characteristic pluripotency, stem cells have the capacity to unveil the mystery behind reproduction, regeneration, and (de)differentiation, rendering stem cell—in particular, embryonic stem cell (ESC)—research essential for the development of a fundamental understanding of biomedical mechanisms and the discovery of clinical therapeutic strategies . HoweveriPSC was initially generated by reactivating nuclear reprogramming factors to reverse differentiated cells into a reprogramming state –8, maintAlthough an increasing number of groups thereafter have made significant efforts in the generation of iPSC from a variety of somatic cell populations, available information about the genome-wide epigenetic alterations that somatic cells must undergo to become fully reprogrammed remains limited. In addition, some concerns about the current procedures, particularly the insufficient efficiency and specificity required for clinical application, remain. Thus, a better understanding of the downstream events following the activation of silenced master reprogramming factors could provide essential information to aid in the development of patient-specific iPSC lines in a faster and safer way. In this review, recent advances in iPSC generation strategies and the detailed mechanisms that underlie reprogramming are highlighted, and future perspectives are discussed.In addition to efficiency and specificity concerns with regard to iPSC generation methods, there has been a concern over the virus based reprogramming as it may integrate unwanted vector fragments into iPSC genome, given that the Yamanaka factors such as Oct4, Sox2, Myc, and Klf4 (OSMK) are introduced into the fibroblast cells with the help of a virus. This would affect the clinical application of derived iPSC as it introduces the possibility of negative effects on the biological properties of iPSC and increases the likelihood of malignant transformation. Indeed, recent study showed that reactivation of viral genes integrated in host genome during differentiation of the reprogrammed iPSC leads to tumorigenesis . To overγ (Cbx3), a protein known to recognize H3K9 methylation, could significantly promote reprogramming [To tackle the problem of low efficiency, chemical as well as epigenetic approaches have been adopted with the aim of enhancing iPSC generation efficiency –12. Epiggramming , 15.Paradoxically, JMJD3, a histone H3K27 demethylase  expectedChanges affecting the dynamic balance between acetylation and deacetylation may also affect reprogramming, as is evidenced by the effects of several core members of nucleosome remodeling and deacetylation (NuRD) repressor complexes on reprogramming efficiency. Serving as a core component in Methyl-CpG Binding Domain Protein 3 (Mbd3) NuRD repressor complexes, Mbd3 can interact with core reprogramming factors (OSKM) and assemble directly with Mbd3/NuRD to recruit the repressor complex to downstream OSKM target genes. As would be expected, Mbd3 depletion was capable of significantly enhancing the reprogramming efficiency of human and mouse fibroblast cells to near 100% within seven days , with thβ receptor II, as is evidenced by the fact that siRNA based knockdown of both targets significantly increases iPSC induction efficiency. Another mechanism for the enhancement of miRNA based reprogramming efficiency is the regulation of cell cycle-related genes [MicroRNA has been acknowledged to function as essential regulators for gene expression in almost all metabolic pathways, suggesting their potential involvement in the regulation of the nuclear reprogramming , and proed genes , 29.Although the native forms of core factors have been widely employed in iPSC generation, their relatively low transactivation activity remains a barrier for somatic cell reprogramming . Recent To enhance iPSC generation efficiency, the Oct4, Sox2, Nanog, and Klf4 (OSNK) reprogramming factors were engineered such that the transactivation domain of YAP is fused to defined factors labeled as OySyNyK. The efficiency of OySyNyK-induced iPSC generation is dramatically enhanced due to these modifications (about 100-fold greater efficiency relative to that of the wild-type OSNK-induced iPSCs). Furthermore, the initiation of reprogramming by OySyNyK is much faster, usually occurring within 24 hours. To understand the mechanism underlying this enhanced reprogramming, an epigenetic study was performed, the results of which indicated that the engineered reprogramming factors significantly increase the expression level of one member, namely, Tet1, of the ten-eleven translocation proteins  at the early reprogramming stage and also produce a marked increase in 5-hydroxymethylcytosine (5-hmC) levels, collectively suggesting that the engineered reprogramming factors collaborate with TETs to regulate 5-hmC mediated epigenetic control of somatic reprogramming .The integration of unwanted vector fragments into the iPSC genome can adversely affect the clinical applications of iPSCs in therapy. Thus, the improvement of nonviral and integration-free alternative methods to eradicate the safety issues currently associated with iPSCs has been a goal since the early stages of iPSC development.The initial approach for the removal of the unwanted viral vector was to combine a lentiviral vector with Cre to excise the reprogramming vectors flanked by loxP sites using transiently expressed Cre-recombinase. Although a large part of the lentiviral vector flanked by loxP sites can be removed, a small part of the vector DNA external to the loxP sites most probably still remains integrated. In addition, this strategy is difficult to operate and time consuming.In another attempt to rectify this issue of viral vector integration, the PiggyBac (PB) transposon system has been used to efficiently integrate the construct harboring core factor genes into TTAA sites in the target genome. The inserted PiggyBac vector can be excised by PB transposase in a footprint-free removal –36.Nonviral methods have also been developed, such as the utilization of episomal and minicircle vectors , 38, adeCompared to other virus vectors such as lentivirus vectors, adenovirus vectors possess many advantages such as conferring the very efficient nuclear entry and low pathogenicity for humans, transducing large genes of more than 30 kb, avoiding integration into the host cell genome, targeting cell specificity, and maintaining long-term expression of transgenes. As such, the adenovirus vectors have been developed as popular gene delivery vehicles in a wide range of transduction for different cell types, particularly for quiescent and differentiated cells in basic biomedical research, clinical applications such as gene therapy, and industrial applications such as vaccine development.Different from all the conventional DNA vectors that so far have been extensively applied, Sendai virus (SeV) vector is a cytoplasmic RNA vector with RNA genome, a material chemically different from the patient's genome DNA. Thus, since the SeV vector replicates its genome exclusively in the cytoplasm instead of entering cell nucleus, it overcomes the fundamental risks in host cell chromosomal alteration caused by integration of DNA vectors into chromosomes or genetic recombination. Additionally, the SeV vector could produce protein in large quantity in the host cells. Given so many advantages over the conventional DNA vectors, SeV has been successfully applied in clinical therapy as well as basic biological research including iPSC generation.To absolutely exclude virus vectors, electroporation of the constructs with nucleofection into somatic cells has become an alternative also .This strategy is based on the administration of engineered mRNA coding for the core reprogramming factors (ONSMK) to avoid innate antiviral responses and has been proven to be significantly more efficient compared to the previously established protocols in reprogramming human fibroblast cells to generate RNA-induced pluripotent stem cells (RiPSCs) . In addiBesides the methods utilizing engineered mRNA  and smalTheoretically, any molecules that target core epigenetic enzymes to alter the dynamic balance of methylation/demethylation could be potential candidates for enhancing or inhibiting somatic reprogramming efficiency . As expeAltogether, recent studies suggest that small molecules which function to alter the dynamic balance between methylation and demethylation—either individually or collaboratively—could significantly enhance reprogramming and largely erase epigenetic memory in cell-specific genes retained in iPSCs, leading to a substantial improvement in the quality of reprogrammed iPSCs , 60, 61.However, its shortcomings cannot be ignored, one of which is that the off-target effects may adversely affect quality of the iPSCs generated.In summary, methods have been developed to remove the unwanted virus vector sequences or exclude use of viruses altogether in virus-free and integration- or transgene-free methods. However, although integration-free methods are within the realm of possibility, these strategies still share the same shortcomings, one of which is extremely low reprogramming efficiency relative to lentiviral vector-mediated strategies. One more issue needs attention for comparison of the reprogramming efficiency mediated by modified mRNA and viral vector strategies such as lentiviral vector. Although the modified mRNA confers the lower induction efficiency relative to the viral vector, the percentage of the normal iPSC generated by this strategy is higher than that by the lentiviral vector due to immune response as well as other adverse factors such as genome instability and chromosomal variation caused by viral vector integration into the chromosome of the iPSC.For large scale generation and further differentiation of iPSCs into special tissues to be used in regeneration medicine for therapeutic purposes, an automatic platform was established starting from fibroblast cell preparation . Since tNot all cells that gain expression of the core reprogramming factors are pluripotent, though they may be self-renewing, because some of them are trapped in a state of partial reprogramming , 48, 63.Modifications of both histones and genomic DNA have been essentially linked to the regulation of master transcription factor-mediated reprogramming. At the histone level, a balance between the enzymes responsible for methylation and those responsible for demethylation, such as methyltransferases and demethylases, determines the global H3K9me3 levels during reprogramming. Genome regions occupied by H3K9me3, a heterochromatic histone mark, have been identified to efficiently prevent the binding of master transcription factors in both human and mouse fibroblasts, serving as major roadblocks during reprogramming –78.Accordingly, Cbx3, a reader of H3K9me3, and H3K9 methyltransferases (Ehmt1 and Ehmt2) have been detected in the Nanog protein complexes in mouse ESCs, suggesting that the Nanog autorepression mechanism is mediated by the recruitment of these readers and methyltransferases by Nanog in the cells remaining in a nonreprogramming state –80.At the genomic DNA level is a situation similar to that at the chromatin level—a balance between DNA methylation and demethylation determines the dynamic status of the somatic cell, leading it towards either reprogramming or remaining differentiated. Again, this balance is also regulated by DNA methyltransferases and demethylases. The demethylation of the promoter regions of the genes conferring pluripotency has become a prerequisite for epigenetic somatic reprogramming  and mostMore and more evidence is available to contribute to an understanding of the mechanisms of passive and active DNA demethylation. However, these two mechanisms do not contribute equally to the dynamic regulation of demethylation. The passive mechanism renders the automatic and gradual loss of methylation during cell cycles possible. In contrast, several enzymes are believed to be involved in the active mechanisms for demethylation, including activation-induced cytidine deaminase (AID) , TET 8484, and tProbably the most important mechanism for active DNA demethylation during induced pluripotency is the conversion of methylcytosine (5-mC) to 5-hydroxymethylcytosine (5-hmC) catalyzed by TET family . Once generated, the 5-hmC has to face several fates, either being directly converted into cytosine (C) through mechanisms involving the base excision repair pathway or sequentially becoming 5-fC and 5-caC to be finally converted into regular cytosine  Figure .Tet1 and Tet2 have been proven to facilitate cell reprogramming , 91. To Although knockdown of all three Tet genes in ESCs seems to confer ESCs normal self-renewal and pluripotency, deletion of Tet1 slightly enhances the reprogramming efficiency  and Tet3The complete abolition of the reprogramming of the Tet TKO MEFs has been linked to failing to undergo mesenchymal to epithelial Transition during MET . Since tApplying the Tet-assisted bisulfite sequencing strategy by which the 5-mC and 5-hmC could be distinguished from each other , it was More interestingly, while both TET1 and TET2 are responsible for the conversion of 5-mC into 5-hmC, their roles in reprogramming are very different, especially in the presence of vitamin C . While T Chlamydomonas, Drosophila, and C. elegans, generating N6-methyldeoxyadenosine (6mA or m6A) [ Drosophila (DMAD) and C. elegans (NMAD-1) function as erasers of 6mA methylation and play essential roles in reproduction and neuronal activities, further suggesting a TET- or TET analogue-mediated epigenetic regulation spectrum in eukaryotes.Besides the methylation of cytosine to form 5-mC, deoxyadenosine methylation has also been detected in the genomes of some eukaryotes, such as or m6A) –99. AlthUnlike TETs, which are guided by the core reprogramming factors and specifically localize to regions of the target genes to enhance active demethylation during reprogramming, some epigenetic regulators, such as macroH2A and peptidylarginine deiminase Padi4, globally rather than specifically affect the pluripotent state either negatively or positively –102. MacIn addition to the genome-wide epigenetic regulation of reprogramming, alterations of key components in some metabolic pathways such as p21 and p53, as well as some general transcriptional and translational apparatuses, also affect reprogramming efficiency. It has been shown that p21 can negatively affect reprogramming efficiency; consequently any alterations which enhance p21 transcription and translation would repress reprogramming . Thus, iThe establishment of iPSC methodology and dissection of the mechanisms regarding reprogramming are a milestone in the long journey of stem cell research both theoretically and practically, providing a sufficient tool with which to tackle fundamental biomedical questions regarding epigenetics-mediated (de)differentiation, as well as providing a valuable cell source for tissue regeneration, human disease modeling, and drug discovery. Due to the efforts made to improve the protocols for iPSC generation, particularly with regard to patient-specific somatic cells, with a focus on increased efficiency and safety, significant progress has been made by employing a variety of (epi)genetic and biochemical approaches. In addition, the molecular mechanisms behind reprogramming have been extensively studied at biochemical, genetic, and epigenetic levels. However, technical challenges in the generation of iPSCs and safety concerns for the use of iPSCs in clinical applications remain big issues which require solving.Although various strategies have been invented to enhance reprogramming efficiency and to improve the issue of safety, regrettably, none of these strategies could ensure both the high generation efficiency and safety of iPSCs. For example, while virus-mediated ectopic expression of the core reprogramming factors or knockdown of key epigenetic factors led to high generation efficiency, the integration of virus vectors could lead to tumorigenesis. Likewise, some strategies without virus vector meditation such as virus vector-free or transgene-free methods, such as the modified mRNA strategy and small molecule treatments to inhibit some barriers or activate enhancers of the reprogramming process, can improve the safety of iPSCs but their efficiency is much lower than that conferred by the virus- or other transgene-mediated methods. In addition, some small molecules may have multiple targets and thus come with the possibility of off-target effects which could lead to the unpredictable quality and safety concerns for the generated iPSCs. Thus, it is of great importance to carry out high-throughput chemical screens, transcriptomic, and proteomic studies so that more small molecules, chromatin remodelers, and other epigenetic modifiers can be identified and employed to enhance iPSC generation efficiency without raising safety and quality issues.On the other hand, mechanism studies may facilitate the discovery of new strategies focusing on different targets to significantly enhance reprogramming efficiency. Previous studies have made this approach applicable, such as those leading to the discovery of epigenetic barriers to the reprogramming. By removing these barriers, reprogramming efficiency could be dramatically promoted. Although several barriers have been identified, more efforts are required to unveil new barriers and activators.Alternatively, microRNA approaches could be also applied for the dissection of pathways implicated in iPSC reprogramming to further understand the crosstalk among metabolic pathways and the molecular agents known to serve as (epi)genetic modifiers or drivers. Further understanding of reprogramming mechanisms and development of safer and more efficient reprogramming strategies will benefit biomedical studies as well as iPSC-mediated regeneration medicine and transplant therapy.


The centrosome is the major microtubule-organising centre of an animal cell. It dictates the arrangement of a radial microtubule array in interphase and bipolar microtubule spindle in mitosis. In differentiated cells, the centrosome also promotes growth of the primary cilium, a sensory antenna-like structure that transduces external stimuli into intracellular signals. The physiological importance of the centrosome is highlighted by the detection of mutations in many genes encoding centrosome components in human developmental disorders, including microcephaly (MCPH), primordial dwarfism (PD), and a range of syndromic ciliopathies .MCPH, a disease characterized by small brain size, and PD, which manifests as whole body growth retardation, are due to reduced cell numbers that result from loss of the relevant stem cell populations. So how do centrosomes contribute to maintenance of stem cell numbers? Centrosomes are structurally complex organelles consisting of two substructures: a centriole pair that is surrounded by pericentriolar material (PCM). The PCM contains proteins responsible for microtubule nucleation and anchoring, while the centrioles provide the scaffold onto which the PCM is assembled. During mitosis centrosomes exhibit increased microtubule nucleating capacity due to PCM expansion. This recruitment of additional PCM components is crucial for spindle assembly and, if it does not occur, disorganised spindles arise that trigger mitotic arrest, cell death and/or aneuploidy. Hence, defective spindle organization in the stem cell niche can trigger cell loss through senescence or apoptosis. However, stem cell number is also determined by spindle orientation with symmetric divisions driving self-renewal and maintenance of stem cell number. Spindle defects that favour asymmetric divisions lead to premature differentiation and reduction of the progenitor pool. Spindle orientation depends upon attachment of astral microtubules from centrosomes to the cell cortex, centrosome defects can disrupt these attachments thereby promoting spindle asymmetry and reduction of the stem cell pool.WDR62 gene), prevents centrosomal recruitment of CDK5RAP2 and γ-tubulin, underlining the importance of centrosome integrity in preventing this disorder [Mutations in two key PCM scaffold components, CDK5RAP2 MCPH3) and pericentrin, cause MCPH and PD, respectively [ and peridisorder . In addiWe have recently described a new regulator of centrosome integrity, the NEK5 protein kinase . NEK5 loLoss of centrosome integrity also contributes to kidney and liver cyst development, a pathological hallmark of many ciliopathies that arise from primary cilium defects. PCM proteins have not been directly linked to ciliopathies, however depletion of pericentrin reduced targeting of intraflagellar transport proteins and polycystin-2 to the centrosome, and inhibited ciliogenesis . Cells c


Diabetic cardiomyopathy (DCM) is a disorder of the heart muscle in people with diabetes that can occur independent of hypertension or vascular disease. The underlying mechanism of DCM is incompletely understood. Some transcription factors have been suggested to regulate the gene program intricate in the pathogenesis of diabetes prompted cardiac injury. Forkhead box transcription factor 1 is a pleiotropic transcription factor that plays a pivotal role in a variety of physiological processes. Altered FOXO1 expression and function have been associated with cardiovascular diseases, and the important role of FOXO1 in DCM has begun to attract attention. In this review, we focus on the FOXO1 pathway and its role in various processes that have been related to DCM, such as metabolism, oxidative stress, endothelial dysfunction, inflammation and apoptosis. Diabetic cardiomyopathy (DCM) is causative for 80 % of the fatality rate in the diabetic inhabitants , 2. The DCM linked pathologies have their ancestries in shifts of gene expression. Although various origins of cardiac injury have been identified, the vital molecular contraptions of their pathogenesis have not been anticipated. The molecular operations within the cardiac cell are organised by transcription factors such as forkhead box (other) transcriptional factor (FOXO). In heart diseases such as atherosclerosis , diabetiDrosophila Melanogaster forkhead mutant and its part was connected to the development of gut of the Drosophila fetus [Drosophila Melanogaster, and abnormal dauer formation-16 in Caenorhabditis elegans [The “forkhead” name was conferred on account of the two spiked-head structures observed in the embryos of the la fetus . Forkheala fetus . FOXO isla fetus , 11. FOX elegans .FOXO’s consists of highly conserved forkhead/winged helix DNA-binding domain, which encompasses the most common 110 amino acids of FOXO family and embodies 3a, 3b, and 2 winged helices, facilitating its DNA binding , 13. FOXOver the recent decade, numerous studies have uncovered the essential functions of FOXOs in managing diverse range of cellular processes. FOXO1 is the key member among the ‘O’ subfamily, in controlling equilibrium of cardiac cells , 30. GloIn relation to the heart, FOXO controls the expression of a variety of target genes that are involved in cellular metabolism, oxidative stress, apoptosis and cell cycle differentiation Table . InteresAmong the FOXO subfamily, FOXO1, FOXO3, FOXO4 are expressed in the heart  and locaDysregulated activity of FOXO1 has been implicated in the pathophysiology of DCM , 7, 68, Diabetic patients are more prone to the risk of cardiomyopathy, and heart failure is a foremost cause of death in diabetes populations –76. ThisDisturbances in myocardial glucose and lipid metabolism are initial events that lead to cardiac dysfunction in diabetic condition. FOXO1 is involved in various pathways related to cellular energy metabolism. During insulin resistance, pyruvate dehydrogenase kinase 4 (PDK4) is known to inhibit glucose oxidation by blocking pyruvate to enter into mitochondrial oxidation through phosphorylating the E1 moiety of pyruvate dehydrogenase complex . EnhanceSurprisingly, Battiprolu and his colleagues noticed morphological and functional myocardial modifications similar to DCM in two dissimilar animal models of type 2 diabetes, db/db mice lacking leptin receptor and high-fat diet obese mice, as an outcome of FOXO boosted pathological alterations. Moreover, mice with cardiac specific FOXO1 knock out were resistant to high-fat diet provoked myocardial dysfunction and hypertrophy . Recent Another cardinal feature of DCM is lipotoxicity resulting in part from excessive lipid accumulation . Puthanvγ, and solute carrier family two (facilitated glucose transporter), member four) in adipocytes, thereby salvaged the diabetic phenotype [Latest studies have disclosed the FOXO1 ability to relate insulin pathway to numerous kinds of metabolic stress in the heart. FOXO1 indirectly controls the insulin sensitivity via negatively modulating the insulin sensing genes. Insulin sensitivity is restored upon FOXO1 restricted deletion in genetic mouse model of insulin resistance, and this conditional deletion further reduced the expression of genes related to gluconeogenesis  in the liver and enhanced insulin sensitizing genes expression  [Stringent regulation of intracellular calcium homeostasis is essential for normal maintenance of cardiac function and growth . Numerouer (NCX) –129. Alter (NCX)  and is ter (NCX) . In addier (NCX) . It has er (NCX) . Surpriser (NCX) . AltogetIn summary, FOXO1 regulation may contribute to the detrimental outcomes of the cardiac cells in diabetes, accelerating the development of DCM, one of the predominant cardiac difficulties in diabetic patients. Metabolic alterations, oxidative stress, endothelial dysfunction, inflammation and apoptosis have been shown to be implicated in the development and progression of DCM, and also in the desirable processes for the regulation of FOXO1 gene. Dysregulated FOXO1 expression and activity appear to promote endothelial dysfunction, myocardial oxidative stress, cardiomyocyte cell death and inflammation observed in DCM. Thus, FOXO1 or, favourably, any of its distinctive pathways may be of extreme concern for pharmaceutical target. However, mechanisms controlling the activity and expression of FOXO1 isoform in DCM are not well appreciated. There are so many unanswered questions relating to the FOXO1 activity in DCM. Some of them like, what could be the mechanistic link whereby FOXO1 activation contributes to the increased vulnerability of diabetic heart to ischemic insults? What could be the cause for persistent activation of FOXO1 in cardiac tissue in the settings of insulin resistance, lipid overload, elevated inflammatory cytokines and hyperglycemia? Whether FOXO1 plays similar roles like insulin resistance, in non-obese type 1 diabetic patients given that insulin deficiency is another characteristic feature of diabetes? Could FOXO1 gene polymorphism be responsible for individual susceptibility of DCM? What could be the direct metabolic consequences of FOXO1 activation? What is the functional role of FOXO1 in nonmyocytes of the heart? Thus, further research is necessary to unveil the precise mechanism of FOXO1 in the development and progression of DCM.


Iliac bone destruction in children is uncommon and presents various imaging features. Correct diagnosis based on clinical and imaging features is difficult. This research aimed to retrospectively explore the clinical features, imaging, and histopathological diagnosis of children with iliac bone destruction. A total of 22 children with iliac bone destruction were enrolled in this retrospective analysis from two children's hospitals during July 2007 to April 2015. Clinical features, imaging, and histopathological findings were analysed. The mode of iliac bone destruction, lesion structure, and the relationship between the range of soft tissue mass and cortical destruction were determined based on imaging data. The data were analysed using descriptive methods. Of the iliac bone destruction cases, eight cases were neuroblastoma iliac bone metastasis, seven cases were bone eosinophilic granuloma, two cases were Ewing's sarcoma, two cases were osteomyelitis, one case was bone cyst, one case was bone fibrous dysplasia, and one case was non-Hodgkin's lymphoma. Iliac bone destruction varies widely in children. Metastatic neuroblastoma and eosinophilic granuloma are the most commonly involved childhood tumours. Bone destruction is a form of bone tissue deletion resulting from the replacement of local bone tissue by pathological tissue; this occurs directly as a result of increased osteoclast activity or indirectly as a result of tumour or inflammation, and both the cortical and cancellous bone may be involved. Bone destruction instances with different causes exhibit different imaging characteristics due to differences in the nature of the diseases, the speed of development, and reactive changes of adjacent bones. The imaging features of iliac bone destruction are not as characteristic as those of the long bone, the lesion type varies widely, and the corresponding imaging features exhibit both diversity and similarity . Hence, In clinical practice, children presenting with unexplained hip pain or lower limp claudication should receive pelvic digit radiography (DR) including the bilateral hip joints and bilateral ilium , 3. The Because iliac bone destruction is uncommon in children, previous studies of iliac bone destruction have often been limited to clinical case reports. Ando et al. reported 3 cases of eosinophilic granuloma of the pelvis in children, and each presented different radiological features and clinical courses . AnotherThis report is a retrospective study that explores the clinical features, imaging results, and histopathological diagnosis of iliac bone destruction in children with the aim of improving the diagnosis of iliac bone destruction in children.This research is a retrospective analysis of 22 children with iliac bone destruction; the children were recruited from the Children's Hospital of SooChow University and the Nanjing Children's Hospital Affiliated to Nanjing Medical University from July 2007 to April 2015. This research was approved by the Ethics Review Board of the Children's Hospital of SooChow University and the Nanjing Children's Hospital Affiliated to Nanjing Medical University. The study was performed in accordance with the ethical standards described in the 1964 Declaration of Helsinki and its later amendments. No conflict of interests existed.Clinical features, imaging, and histopathological findings were analysed to improve the diagnostic level of iliac bone destruction in children. The inclusion criteria were as follows: (1) a child patient presenting with hip pain or lower extremity claudication as the initial symptom, and X-ray images showing iliac bone destruction and (2) a routine pelvic radiograph examination in a child patient showing iliac bone destruction without abnormal pathological changes in other organs. The exclusion criteria were as follows: (1) a child patient with previous hematopoietic malignancy, abdominal malignant tumour or soft tissue malignant tumour, and iliac bone destruction; (2) a child patient with iliac bone destruction after suffering severe iliac trauma; and (3) a child with postoperative recurrence of iliac bone destruction with known pathologic diagnosis. The pelvic DR of these patients included the bilateral hip joints and bilateral ilium. Laboratory examinations, including routine blood tests, erythrocyte sedimentation rate (ESR), and the level of alkaline phosphatase, were also recorded. If the pelvic radiograph indicated iliac bone destruction, the patient received a CT or MRI examination to further verify the diagnosis of iliac bone destruction. All patients were diagnosed with iliac bone destruction according to X-ray and CT/MRI examinations were confirmed by operation and histopathological examination. Finally, data were analyzed using descriptive methods.Among the 22 patients, the age of onset ranged from 11 months to 12 years , and 14 (63.6%) of the cases were in the 2- to 5-year-old age range. In the studied cases of iliac bone destruction, 6 were on the left side, 13 were on the right side, and 3 were on both sides; 19 cases were found in the iliac ala, and 3 were found in the acetabulum. The study included 15 males and 7 females, with an age range from 11 months to 12 years . The course of the disease ranged from 2 days to 1 month, with an average of 7 days. Among the cases, nine patients were diagnosed with hip joint pain as the first symptom, five were diagnosed with lower limb claudication, four were diagnosed with hip pain and fever, two were diagnosed with backache and malaise, and two were diagnosed unintentionally when X-ray films were recorded for lower limb trauma. Increased ESR was observed in five cases, decreased haemoglobin (lower than 110 g/L) was observed in six cases, increased white blood count was observed in four cases, and elevated alkaline phosphatase levels were observed in four cases.The imaging feature common to all 22 cases was iliac bone destruction of various forms; 8 cases were accompanied by soft tissue masses of varying sizes, and 2 cases exhibited needle periosteal reaction. Pathology results showed seven disease types: 8 cases of neuroblastoma iliac bone metastasis, 7 cases of bone eosinophilic granuloma, 2 cases of Ewing's sarcoma, 2 cases of osteomyelitis, 1 case of bone cyst, 1 case of bone fibrous dysplasia, and 1 case of non-Hodgkin's lymphoma.Eight patients with neuroblastoma iliac bone metastasis were from 2 to 6 years old . An X-ray examination showed osteolytic bone destruction without a clear destruction edge in 6 cases and a soft tissue mass in the destruction zone close to the edge of the iliac bone in three cases. Of the 6 patients who received CT scans, 5 showed mixed bone destruction, primarily iliac osteolytic bone destruction; 1 showed osteolytic bone destruction; and 2 showed needle periosteal reaction around the area of bone destruction. Of the three patients who received an MRI examination, the intense signal of fat in the iliac bone marrow cavity was replaced by a weak signal, and soft tissue masses were observed around the lesion, primarily with equal T1 and equal T2 signals, which were uneven.Seven patients having bone eosinophilic granuloma were from 2 to 8 years old . The osteolytic bone destruction in these patients was associated with a soft tissue mass shadow of uneven density; the edge was clear, and sclerosis manifestation was not seen. X-ray and CT examinations showed varying degrees of irregular iliac osteolytic destruction; the edge was partly clear, and a soft tissue mass shadow was obvious, with a sclerotic edge in one case. CT examination showed expansive and osteolytic iliac bone destruction zones, an obvious soft tissue mass around the iliac bone, and blurring of the sacroiliac joint interspace .Both cases of Ewing's sarcoma were in the ilium ala with local irregular osteolytic bone destruction; an obvious medullary cavity sclerosis was observed in one case. Part of the cortical bone was destroyed and absent with rough edges. A large soft tissue mass was seen surrounding the front and back of the abnormal ilium. The ipsilateral sacroiliac joint and hip joint were not involved.In both cases of osteomyelitis, hemilateral widespread irregular bone destruction was observed with reduced and uneven density, trabecular bone blurring, and varying degrees of sclerotic bone surrounding the lesion. Obvious hyperplasia and sclerosis were often observed around the bone destruction caused by chronic osteomyelitis.One 12-year-old boy had a bone cyst. Single-cystic expansive bone destruction was seen in his right ilium with expansion of the local cortical bone, a clear hardening edge, and discontinuous cortical bone .One 11-year-old boy had bone fibrous dysplasia. The bone lesion was of the cystic destruction type and presented as an oval-shaped change with a clear boundary, a hardened edge, and expansively thinning bone cortex.In one patient having non-Hodgkin's lymphoma, the ilium was extensively changed. X-ray and CT examinations of this patient showed worm-eaten osteolytic invasive bone destruction with uneven density around the destruction zone and unclear edge. A mild sclerosis reaction, adjacent soft tissue swelling, and soft tissue mass were observed, and the fat space was blurred. MRI showed T1-weighted images of intermediate signal and T2-weighted images of high signal change in the iliac destruction region and in the muscle around the ilium. The immunophenotype of this case was large B-cell lymphoma.The clinical symptoms of iliac bone destruction in children lack common characteristics. In all 22 cases enrolled in this study, hip pain and lower limb claudication were the most common first symptoms, and a few cases showed fever associated with the first symptom. The course of the disease varied from 2 days to a month, with an average of 7 days. The age of patients mainly ranged from 2 to 5 years . Hip pain was mainly experienced as intermittent dull paint hat was intensified after exercise and relieved after rest, presenting an intermittent attack form [Iliac bone destruction is a form of bone tissue deletion that results from the replacement of local bone tissue by pathological tissue, which occurs directly as the result of increased osteoclast activity or indirectly as the result of tumour and inflammation, and both cortical and cancellous bone may be involved. Bone destruction instances with different causes exhibit different imaging characteristics, due to the differences in the nature of the diseases, the speed of development, and reactive changes of adjacent bones. The imaging features of iliac bone destruction in children are not as characteristic as those of the long bone, the lesion type varies widely, and the corresponding imaging features exhibit both diversity and similarity . Bone deBenign lesions causing iliac bone destruction in children also presented as osteolytic destruction, as with the case of bone eosinophilic granuloma  observedChildren presenting with unexplained hip pain or lower limb claudication should receive a pelvic X-ray, including the bilateral hip joints and bilateral ilium , 3. If tOf the 22 patients with iliac bone destruction, most experienced varying degrees of clinical or radiological misdiagnosis before operation, with the exceptions of one case of osteomyelitis, one case of bone cyst, and three cases of bone eosinophilic granuloma, for which the preoperative diagnoses were more precise. Most of the preoperative misdiagnoses were actually osteomyelitis . Seven dFrom a pathological viewpoint, the primary lesion of neuroblastoma lacks specific symptoms and signs and presents a high degree of malignancy. Early metastasis occurs very often in patients with this disease. In 60% of child patients, metastasis has already occurred at the time when they first see a doctor; the most common metastasis sites are the bone and marrow . BecauseBased on this retrospective analysis of the cases studied here, it can be concluded that neuroblastoma iliac bone metastasis and bone eosinophilic granuloma are the most common forms of iliac bone destruction in children; Ewing's sarcoma, osteomyelitis, bone cyst, bone fibrous dysplasia, and non-Hodgkin's lymphoma are relatively rare . In clinIliac bone destruction in children is comparatively uncommon. This disease type has some characteristic imaging features, but, generally, nonspecific imaging findings were found. A histopathological examination must be considered when making a diagnosis. Neuroblastoma iliac bone metastasis and bone eosinophilic granuloma are the most common forms of iliac bone destruction in children; Ewing's sarcoma, osteomyelitis, bone cyst, bone fibrous dysplasia, and non-Hodgkin's lymphoma are relatively rare.


Altered sensory experience in early life often leads to remarkable adaptations so that humans and animals can make the best use of the available information in a particular environment. By restricting visual input to a limited range of orientations in young animals, this investigation shows that stimulus selectivity, e.g., the sharpness of tuning of single neurons in the primary visual cortex, is modified to match a particular environment. Specifically, neurons tuned to an experienced orientation in orientation-restricted animals show sharper orientation tuning than neurons in normal animals, whereas the opposite was true for neurons tuned to non-experienced orientations. This sharpened tuning appears to be due to elongated receptive fields. Our results demonstrate that restricted sensory experiences can sculpt the supranormal functions of single neurons tailored for a particular environment. The above findings, in addition to the minimal population response to orientations close to the experienced one, agree with the predictions of a sparse coding hypothesis in which information is represented efficiently by a small number of activated neurons. This suggests that early brain areas adopt an efficient strategy for coding information even when animals are raised in a severely limited visual environment where sensory inputs have an unnatural statistical structure. The perceptual and cognitive abilities of humans and animals often develop by adapting to a particular environment2345678910112345678How does the primary visual cortex deal with such redundancy? One potential solution to this problem is sharpening the orientation tuning of individual neurons; if the orientation tuning curves of single neurons are sharpened to cover a very limited range, such an adaptation allows different sets of neurons to be activated for different orientation stimuli even in a restricted orientation range. In fact, a computational study suggested that they shouldOrientation restriction provides an opportunity to evaluate quantitatively how structured visual experiences influence the functional development of single neurons in a manipulated environment. To guarantee that the animals always experienced a particular orientation, we fabricated goggles with cylindrical lenses +67 diopter) that allow optical patterns to be transmitted in a severely restricted orientation range 7 diopterTo examine the distribution of preferred orientations in the early visual cortex, optical imaging based on intrinsic signals6To study the effect of orientation-restricted rearing on various tuning properties of individual neurons, single unit activities were recorded using a pair of tungsten microelectrodes inserted into the primary visual cortex of anaesthetised and paralysed cats. Seventeen penetrations were made oblique to the cortical surface in three v-goggled cats. For comparison, the same set of data was also collected from three age-matched normal cats (17 oblique penetrations). Receptive field profiles are closely related to the tuning properties for orientation and spatial frequency15161718192022et al.When the length-to-width ratio of receptive fields was plotted against the recorded depth, no relationship was found for neurons that were tuned to the vertical orientation in v-goggled cats (data not shown). In addition, none of the tuning parameters described in the next subsection appeared to have a relationship with cortical depth. Thus, the functional properties of single neurons examined in this study  were not different across cortical layers  receptive field elongated along its preferred orientation has sharper tuning than a non-elongated, circular one1516181920To test this, neuronal responses were examined by presenting gratings of various orientation and spatial frequency combinations23In v-goggled cats, neurons tuned to the vertical orientation generally preferred a similar spatial frequency . Orientation selectivity appeared to be sharpened, at least partly, by the elongation of receptive fields for neurons tuned to the experienced orientation. In addition to the well-appreciated increase in the proportion of neurons tuned to the experienced orientation23456785Although other possibilities cannot be ruled out, the above results are highly consistent with a theoretical prediction2829Here, we demonstrate that the primary visual cortex adapts to an orientation-restricted environment by sharpening the orientation tuning of the majority of single neurons, while allocating other cells to non-experienced stimuli in an economical manner through sparse coding13282916173132et al.et al.et al.35This is not the first investigation to examine the orientation bandwidth of single neurons in orientation-restricted animals. Sengpiel et al.23456782345678The majority of neurons in early sensory areas develop preferences to the exposed stimulus when young animals are reared in an environment dominated by a particular sensory stimulus2345678910112345678373839234567893738394041424445All animal care and experimental guidelines were in accordance with those established by the US National Institutes of Health and were approved by the Institutional Animal Research Committee at RIKEN  and by the Osaka University Animal Care and Use Committee . The experiments were conducted on anaesthetised and paralysed cats.For orientation restriction, we fabricated goggles with cylindrical lenses +67 diopters) so that optical patterns could be transmitted in a severely limited orientation range 6 diopters6The procedures for animal preparation and maintenance, surgery, and experiment setup for optical imaging based on intrinsic signals have been described in detail elsewhere65The procedures for animal preparation and maintenance, surgery, single-unit recording, and experiment setup have been described in detail elsewhere21To map receptive fields, a small optimal grating was presented at various positions in rapid succession . The diameter of these gratings was equal to 3 sampling points and was customised for each neuron to maximise both the responses and spatial resolution. The obtained maps were fitted with a 2D Gaussian function to determine the spatial extent of the receptive fields at the 10% peak value. Length was measured along the directions of the preferred orientations determined below, whereas width was measured along their orthogonal directions.To examine orientation and spatial frequency tunings, sinusoidal gratings of various orientation and spatial frequency combinations were presented in rapid succession 23To compare stimulus selectivity across neuronal groups, the Tukey-Kramer method was used to correct for multiple comparisons after the Kruskal-Wallis test. In these tests, medians were compared across 3 neuronal groups: neurons tuned to the vertical orientation in v-goggled cats, neurons tuned to non-vertical orientations in v-goggled cats, and all neurons in normal cats.How to cite this article: Sasaki, K. S. et al. Supranormal orientation selectivity of visual neurons in orientation-restricted animals. Sci. Rep.5, 16712; doi: 10.1038/srep16712 (2015).


NH4+-N concentration and SDD/WL were the most important controlling factors for VPF. Therefore, increased anthropogenic nutrient inputs and a degraded underwater light climate surely result in a decreased VPF. These results elucidate the driving mechanism of aquatic vegetation degradation and will facilitate Lake Taihu ecological restoration.Terrestrial and aquatic ecosystem degradation is widely recognized as a major global environmental and development problem. Although great efforts have been made to prevent aquatic ecosystem degradation, the degree, extent and impacts of this phenomenon remain controversial and unclear, such as its driving mechanisms. Here, we present results from a 17-year field investigation (1998–2014) of water quality and a 12-year remote sensing mapping (2003–2014) of the aquatic vegetation presence frequency (VPF) in Eastern Lake Taihu, a macrophyte-dominated bay of Lake Taihu in China. In the past 17 years, nutrient concentrations and water level (WL) have significantly increased, but the Secchi disk depth (SDD) has significantly decreased. These changes were associated with increased lake eutrophication and a degraded underwater light climate that further inhibited the growth of aquatic vegetation. In Eastern Lake Taihu, increased nutrients, chlorophyll  Aquatic vegetation supports critical ecological services by providing the habitat for a diverse and economically important faunal community, sequestering carbon and nutrients, stabilizing sediment and shorelines121157in situ and hyperspectral multispectral infrared and visible imaging spectrometer (MIVIS) images acquired over a 13-year period in Lake Garda  indicated that nearly 98% of macrophyte meadows were lost and subsequently replaced by moderate to extremely sparse communities with densities from 10% to 40% and that well-established submerged macrophytes (including Charophytes) were replaced by opportunistic aquatic plant speciesUnfortunately, the aquatic vegetation of lake, coastal and marine ecosystems has undergone substantial degradation with the onrushing advance of human settlement and water resources exploitation throughout the world in past decades191011e.g., reduced SDD and increased phytoplankton, hypoxia, and anoxia in surficial sediments710151617These changes have largely been attributed to eutrophication caused by increasing nutrients and sediments from the alteration of the surrounding catchments and their subsequent effects on water quality, In China, many shallow lakes have also undergone rapid aquatic vegetation degradation and water quality deterioration. The degradation process of aquatic vegetation is highly complex, and several affecting factors are often interlinked, making it important to consider both natural variation and anthropogenic effects. Many previous studies have tracked the spatio-temporal variability of the distribution of aquatic vegetation in shallow lakes and have explored the potential effects of environmental factors on aquatic vegetation using field investigations, experimental cultures, remote sensing and ecological modeling1920Aside from conventional techniques, such as field mapping and photography, remote sensing can be an effective tool for mapping aquatic vegetation distributions over large scales. Many remote sensing techniques and methods, including spectral inversion, classification trees, the normalized difference vegetation index (NDVI), the floating algae index (FAI) and vegetation presence frequency (VPF), have been developed to accurately identify aquatic vegetation using different types of remote sensing imagery 2223242526276in situ aquatic vegetation to satellite remote sensing mapping had not been discussed in this study because they were beyond the scope of this manuscript; furthermore, these topics had been addressed in our previous studyTherefore, the purpose of this study was to combine long-term water quality monitoring data and remote sensing mapping of aquatic vegetation to elucidate the mechanisms by which increased eutrophication and a degraded underwater light climate had impacted environmental quality and thus degraded the aquatic vegetation in Eastern Lake Taihu, a shallow macrophyte-dominated bay located in the southeastern portion of Lake Taihu. We first explored the long-term dynamics of the habitat environment of the aquatic vegetation. We then used multi-year MODIS data to systematically quantify the interannual characteristics of the situation and strength of aquatic vegetation and the associated variability. Finally, we attempted to identify quantitative links between the responses of aquatic vegetation to increased eutrophication and the deteriorated underwater light climate. However, calibration, validation and quantitative comparisons of 2, a mean depth of 1.9 m and a catchment area of 36,900 km2 1530Lake Taihu, the third largest freshwater lake in China, is located in the Yangtze River Delta and has a water surface area of 2,338 km2 and a mean water depth of 1.2 m.Eastern Lake Taihu  is a shallow macrophyte-dominated bay located in the southeastern portion of Lake Taihu . AccordiA 17-year (1998–2014) program of long-term site-specific observations has been conducted in Eastern Lake Taihu , •. One 4+-N), total phosphorus (TP), dissolved total phosphorus (DTP), and chlorophyll a (Chla). Samples for the determination of TSM, TDN, TDP, and Chla concentrations were filtered using Whatman GF/F filters. Chla was extracted with ethanol (90%) at 80 °C and analyzed spectrophotometrically at 665 nm and 750 nm using a Shimadzu spectrophotometer with a correction for phaeopigments (Pa)μm. Samples were then dried (105 °C for 4 h) and weighed to calculate the TSM concentration.The habitat environment of aquatic vegetation includes many aspects, such as water level (WL) (a substitute of water depth), nutrients, light availability, sediment, and predation pressure. Here, our study used observational data from TLLER to define critical SAV habitat requirements in terms of nine aquatic environmental and water quality variables: WL, SDD, total suspended matter (TSM), total nitrogen (TN), dissolved total nitrogen (DTN), ammonia . NH4+-N concentration was measured using a flow injection analyzer .Samples were analyzed for the concentrations of five nutrients—TN, TDN, NHThe daily WL of Lake Taihu was recorded at Jiapu and Xiaomeikou , □. The We used the FAI methodj exceeded the FAI threshold of −0.025, the value of this pixel was set to 1 in the FAI layer; otherwise, the value of the pixel was set to 0. The VPF in each pixel was then evaluated in a given set of n images:When the FAI of pixel j in the image set of n and represents the proportion of pixel j with FAI = 1 in the total number of images (n). A detailed description of VPF is provided in our previous studywhere VPF is the vegetation presence frequency of the pixel p < 0.05). The significance levels were reported as significant (p < 0.05) or non-significant (p > 0.05).Statistical analyses, including calculations of the average, maximum, and minimum values and linear and non-linear regressions, were performed using Statistical Program for Social Sciences (SPSS) 17.0 software. We performed correlation analyses using SPSS software to investigate the relationships between the variables. The differences in the parameters were assessed by one-way analysis of variance (ANOVA)  provided a flexible and effective technique for modeling nonlinear time-series dataset35i.e., we assessed the amount of variation explained by each predictor based on the multiple linear regression. TSM, TN, DTN, NH4+-N, TP, DTP, and Chla were used as the predictors of relative importance. In addition, aquatic vegetation was largely determined by the light availability of the lake bottoma) according to the variance inflation factors (VIFs), and 10 was used as a threshold value4+-N, TP, DTP) and then used the metric “lmg” with relaimpo package in R version 3.0.0R2 into nonnegative contributions for each predictor termR2R2 but removes the dependence on orderings that bias stepwise regression by averaging over orderings, using simple unweighted averagesR2 among predictors because it uses both indirect and direct effects and adjusts for other predictors in the overall model41To determine the relative importance of environmental variables on the VPF, we used the concept of dispersion importance, 4+-N, TP, DTP, Chla, and SDD/WL were used as the predictors of the CART model.In addition, the relationship between VPF and habitat environment factors could be strongly non-linear and involved complex interactions that could not completely be explained by previously used linear statistical modeling approaches. The classification and regression tree (CART) model, as an efficient tool for extracting key variables and thresholds within a multivariate dataset, has been widely used in a variety of fields, such as environment and ecology434+-N, and Chla exhibited a nearly linear increasing trend , partially reflected the life history characteristics of the aquatic vegetation in Eastern Lake Taihu. In winter, most areas of the bay had a VPF below 0.5  . The relative contributions of the variables to explaining the total variance were SDD/WL > NH4+-N > TN > DTP > TP > TSM. Therefore, SDD/WL and NH4+-N were the two most important variables, explaining 60.12% of the total R2 of the multiple linear regression.Because the nine variables are inter-correlated , differeand DTN)  through N = 12 years, data = 100%)  further based on the high and low NH4+-N concentration conditions. During the low NH4+-N concentration condition, the ratio of SDD/WL was another important factor that affected VPF in Lake Taihu. A significant increase in SDD/WL (>0.1447) resulted in the high VPF condition  at this node relative to Node 3. In summary, high NH4+-N concentration, high SDD/WL and low SDD/WL controlled VPF by approximately 33.3%, 25.0% and 41.7%, respectively, in Eastern Lake Taihu from 2003 to 2014. The CART model showed that NH4+-N and SDD/WL were the most important factors controlling VPF variability in Eastern Lake Taihu, which was consistent with the results of the relative contributions of the individual variables to the VPF.In addition to the analysis of the relative importance based on the linear relationship excluding the effects of auto-correlated variables, the non-linear CART model was used to extract key variables and thresholds in this study. The final CART model showed that a root node (Node-1) was a starting point that included all data ( = 100%) . NH4+-N It is increasingly recognized that efforts to understand ecological resilience and facilitate the management and recovery of aquatic vegetation must ascertain the large-spatial-scale and temporal dynamics rather than considering only simple station data and static cover types4748242223242526Our study used the VPF method based on MODIS data to map the changes in distribution patterns of aquatic vegetation in Eastern Lake Taihu. Frequently repeated satellite image acquisition, in combination with long-term site-specific observations of water quality, has a clear advantage for discussing the temporal and spatial dynamics and potential controlling factors of aquatic vegetation relative to other mapping techniques. However, our study shows only that changes in aquatic vegetation cover have occurred and cannot identify the community succession of aquatic vegetation because a pixel size and mapping unit less than 10–15 m would have been required to accurately map species compositionPrevious studies have demonstrated the link between water environment conditions and aquatic vegetation growth in experimental, observational and remote sensing studies in numerous locations worldwide455220264+-N concentration and SDD/WL were the two most important variables related to VPF; therefore, the changes in the aquatic vegetation of Eastern Lake Taihu were primarily controlled by eutrophication (nutrients and biomass) and hydrologic conditions (WL and underwater light climate). These findings, combined with our linear regression results, imply that the increased trophic level in Eastern Lake Taihu enhanced phytoplankton biomass. Both increased WL and decreased SDD caused by the increase in phytoplankton biomass and TSM resulted in decreases in the ratio of euphotic depth to water depth and further bottom darkening in this area. Healthy and environmentally harmonious values of this ratio are vital to the growth and distribution of aquatic vegetation and even the shift from macrophyte to phytoplankton dominance39The relative importance analysis and CART model conducted in our study indicated that NHCompared with phosphorus concentration, nitrogen concentration, particularly ammonia concentration, contributed more to the degradation of aquatic vegetation in Eastern Lake Taihu , Table 35556In addition, although we have no long-term observation data of sediment, we did not neglect the potential important roles of lake sediments in driving the nutrient cycle and degradation of SAV in shallow lakes. Sediment is considered an important nutrient source for rooted aquatic vegetation, and fertile sediment plays a positive role in the growth, development, reproduction and germination of aquatic vegetation220a concentration and to the decreased SDD in Eastern Lake Taihu. The identification of the controlling factors of aquatic vegetation in Eastern Lake Taihu will help the manager make decisions for recovering aquatic vegetation. In addition, the empirical relationships and CART model developed in this study can be used to assess and forecast the effects of changes in eutrophication and light availability on the status of aquatic vegetation and to help define restoration targets.To determine the specific and practically feasible management measures to preserve and restore aquatic vegetation, the quantitative links between the habitat environment and vegetation response must be identified. Our study showed that the habitat environment of aquatic vegetation deteriorated continuously and that the VPF was significantly correlated with intensified eutrophication due to the increase in nitrogen and phosphorus concentrations and deteriorated underwater light environment due to the increased WL, TSM, and ChlOur study showed that nitrogen concentration overrided phosphorus concentration in the degradation of aquatic vegetation in Eastern Lake Taihu, which provided new evidence for nitrogen reduction for aquatic vegetation recovery and eutrophication control. Previous nutrient loading analyses, nutrient addition bioassays and nutrient cycling studies showed that both nitrogen and phosphorus input reductions were required for long-term eutrophication and cyanobacterial bloom control in Lake Taihu59Because the shift from a macrophyte-dominated clear water ecosystem to a phytoplankton-dominated turbid water ecosystem typically lacks massive, marked early-warning signals, special attention should be given to precipitating events rather than to the underlying loss of resilience. Indeed, once eutrophication begins to impact aquatic vegetation, a self-perpetuating macrophyte-dominated clear water ecosystem can be visualized. Many stochastic events that are typically difficult to predict or control can easily trigger state shifts In addition, the significant negative correlation between WL and VPF and the positive correlation between SDD and VPF, and SDD/WL as the key controlling factor of VPF indicated that the improvement of underwater light availability was also crucial to the growth, distribution and recovery of aquatic vegetation in many turbid shallow waters3953524+-N concentration and SDD/WL were the most important controlling factors of VPF based on the relative importance partition analysis and CART model.This study demonstrates the changes in aquatic vegetation in response to increased eutrophication and deteriorated underwater light climate in Eastern Lake Taihu by combining long-term site-specific observations of water quality and temporal-spatial patterns of aquatic vegetation from remote sensing mapping. Our study indicates that several water quality parameters associated with aquatic vegetation growth have deteriorated from 1998 through 2014 in Eastern Lake Taihu due to eutrophication, which resulted in the degradation of aquatic vegetation. Of all habitat environment factors considered in this study, NHAlthough many controlling measurements have been implemented to reduce eutrophication and some monitoring data have showed nutrient load reductions in many inflowing river of Lake Taihu since the implementation of the Major Projects on Control and Rectification of Water Body Pollution in 2008, variable nutrient sources and internal cycles have not markedly improved the lake water quality of Eastern Lake Taihu. Continued population growth, economic development around Lake Taihu and the stressors of climate warming will likely hinder the effectiveness and success of these projects. However, the strong negative correlation between the VPF and nutrient concentrations suggests that further nutrient reductions will be necessary for aquatic vegetation restoration in Eastern Lake Taihu. Examples of areas where water quality has recovered in a holistic manner are needed to facilitate and encourage restoration efforts in Lake Taihu. In addition, lowering the WL to increase SDD/WL and underwater light availability is a potential method of promoting aquatic vegetation recovery in large shallow lakes.How to cite this article: Zhang, Y.L. et al. Aquatic vegetation in response to increased eutrophication and degraded light climate in Eastern Lake Taihu: Implications for lake ecological restoration. Sci. Rep. 6, 23867; doi: 10.1038/srep23867 (2016).