Barriers to and facilitators of success for early and Mid-Career professionals focused on bipolar disorder: A global needs survey by the International Society for Bipolar Disorders.

INTRODUCTION: The International Society for Bipolar Disorders created the Early Mid-Career Committee (EMCC) to support career development of the next generation of researchers and clinicians specializing in bipolar disorder (BD). To develop new infrastructure and initiatives, the EMCC completed a Needs Survey of the current limitations and gaps that restrict recruitment and retention of researchers and clinicians focused on BD. METHODS: The EMCC Needs Survey was developed through an iterative process, relying on literature and content expertise of workgroup members. The survey included 8 domains: navigating transitional career stages, creating and fostering mentorship, research activities, raising academic profile, clinical-research balance, networking and collaboration, community engagement, work-life balance. The final survey was deployed from May to August 2022 and was available in English, Spanish, Portuguese, Italian, and Chinese. RESULTS: Three hundred participants across six continents completed the Needs Survey. Half of the participants self-identified as belonging to an underrepresented group in health-related sciences (i.e., from certain gender, racial, ethnic, cultural, or disadvantaged backgrounds including individuals with disabilities). Quantitative results and qualitative content analysis revealed key barriers to pursuing a research career focused on BD with unique challenges specific to scientific writing and grant funding. Participants highlighted mentorship as a key facilitator of success in research and clinical work. CONCLUSION: The results of the Needs Survey are a call to action to support early- and midcareer professionals pursuing a career in BD. Interventions required to address the identified barriers will take coordination, creativity, and resources to develop, implement, and encourage uptake but will have long-lasting benefits for research, clinical practice, and ultimately those affected by BD.

Salvianolic acids from Salvia miltiorrhiza Bunge and their anti-inflammatory effects through the activation of α7nAchR signaling.

ETHNOPHARMACOLOGICAL RELEVANCE: Cardiovascular disease (CVD) is a serious disease with a high incidence rate and mortality. Inflammation is closely related to the occurrence of CVDs. As an essential medicine of promoting blood circulation and removing blood stasis in China, Salvia miltiorrhiza Bunge (Danshen) is widely used to treat CVDs due to its anti-inflammatory and cardiovascular protective effects. Salvianolic acids are the most abundant component in the water extract of S. miltiorrhiza, which has a significant effect on the treatment of CVDs. However, due to the complex composition of salvianolic acids, the active molecules and their underlying mechanisms have not been fully explored. AIM OF THIS STUDY: The present study aims to isolate and identify salvianolic acids from Danshen with anti-inflammatory activity and explore the potential mechanisms of isolates. METHODS: The structures of isolated salvianolic acids were elucidated by UV, IR, NMR, MS and electronic circular dichroism (ECD) calculations. Then anti-inflammatory activities of isolates were screened out by the zebrafish inflammation models. The most active compound was further used to explore the anti-inflammatory mechanisms on LPS-stimulated RAW 264.7 cells. The key inflammatory cytokines IL-6 and TNF-α were measured by enzyme-linked immunosorbent assay (ELISA). The protein expression levels of STAT3, p-STAT3 (Tyr705), NF-κB p65, IκBα, p-IκBα (Ser32) and α7nAchR were determined by Western blotting. The nuclear translocation of p-STAT3 (Tyr705) and NF-κB p65 was evaluated by immunofluorescence assays. Finally, the in vivo anti-inflammatory mechanisms were investigated by observation of neutrophil migration, H&E staining, survival analysis and quantitative PCR (Q-PCR) in LPS-microinjected zebrafish. RESULTS: Two new and four known compounds were isolated from Danshen. Among them, isosalvianolic acid A-1 (C1) and ethyl lithospermate (C5) inhibited neutrophil migrations in three zebrafish inflammation models and C1 with the best activities decreased the secretion of IL-6 and TNF-α and inhibited the expression level of p-IκBα (Ser32) in LPS stimulated RAW 264.7 cells. In addition, C1 also reduced the nuclear translocation of NF-κB p65 and p-STAT3 (Tyr705). Moreover, C1 significantly upregulated the protein expression of α7nAchR, and the knockdown of α7nAchR counteracted the effects of C1 on the production of IL-6 and TNF-α and the expression levels of p-STAT3 (Tyr705), NF-κB p65 and p-IκBα (Ser32). In vivo experiments, C1 decreased the migration and infiltration of inflammatory cells, increased the survival ratio and inhibited the mRNA level of IL-6, TNF-α, STAT3, NF-κB and IκBα in LPS-microinjected zebrafish. CONCLUSION: Two new and four known compounds were isolated from Danshen. Among them, C1 exerted anti-inflammatory activities by activating α7nAchR signaling and subsequently inhibiting STAT3 and NF-κB pathways. This study provided evidence for the clinical application of Danshen and contributed to the development of C1 as a novel in the treatment of cardiovascular disease.

Efficacy and safety of angiogenesis inhibitors plus immune checkpoint inhibitors in advanced soft tissue sarcoma: a real-world, single-center study.

Angiogenesis inhibitors (AIs) and immune checkpoint inhibitors (ICIs) are new treatment options for advanced soft tissue sarcoma (STS) patients. This study evaluated the efficacy and safety of AIs plus ICIs in patients with advanced STS. A retrospective cohort study was performed on STS patients treated with AIs and ICIs at Shandong Cancer Hospital and Institute between August 2020 and December 2021. The primary endpoint was objective response rate (ORR); secondary endpoints included progression-free survival (PFS), disease control rate (DCR), overall survival (OS), and adverse events. Thirty-three patients were enrolled; 27 were evaluable for objective response. The ORR and DCR were 48.1% (95% CI 30.7-66.0%) and 85.2% (95% CI 67.5-94.1%). With a median follow-up of 7.6 months (range, 0.8-25.5), the median PFS for all 33 patients was 8.90 months (95% CI 5.98-11.82). The median OS was not reached. The most common treatment-related adverse events (TRAEs) of any grade were hypertension (50.0%), ECG T-wave abnormality (30.0%), hypothyroidism (26.7%), elevated alanine aminotransferase or aspartate aminotransferase (23.3%), elevated thyroid-stimulating hormone (23.3%), and fatigue (16.7%). The most common grade 3-4 TRAE was hypertension (27.3%). Three serious TRAEs (two myocarditis and one rapid atrial fibrillation) were recorded. This study suggests that adding AIs to ICIs is beneficial in STS.

Anti-inflammatory, anti-angiogenetic and antiviral activities of dammarane-type triterpenoid saponins from the roots of Panax notoginseng.

Panax notoginseng has been used both as a traditional medicine and as a functional food for hundreds of years in Asia. However, the active constituents from P. notoginseng and their pharmacologic properties still need to be further explored. In this study, one new dammarane-type triterpenoid saponin (1), along with fourteen known analogs (2-15) were isolated and identified from the roots of P. notoginseng. The anti-inflammatory, anti-angiogenetic and anti-dengue virus effects of these isolated compounds were further evaluated. Compounds 1, 3, 5-7 and 10-12 exerted anti-inflammatory effects in two different zebrafish inflammatory models. Among them, 11, with the most significant activities, alleviated the inflammatory response by blocking the MyD88/NF-κB and STAT3 pathways. Moreover, compound 15 showed anti-angiogenetic activities in Tg(fli1:EGFP) and Tg(flk1:GFP) zebrafish, while 3 and 5 only inhibited angiogenesis in Tg(fli1:EGFP) zebrafish. Additionally, compounds 1, 3, 6, 8, 9 and 12 suppressed the replication of dengue virus either at the viral adsorption and entry stages or at the intracellular replication step. In conclusion, these findings enrich knowledge of the diversity of saponins in P. notoginseng and suggest that the dammarane-type triterpenoid saponins from P. notoginseng may be developed as potential functional foods to treat inflammation, angiogenesis or dengue-related diseases.

Effect of lithium on circadian rhythm in bipolar disorder: A systematic review and meta-analysis.

OBJECTIVES: Circadian rhythm disruption is commonly reported in patients with bipolar disorder. Lithium has been suggested to have effects on the circadian clock, the biological basis of the circadian rhythm. The objective of the current review was to review systematically the existing studies on the effect of lithium on circadian rhythm in patients with bipolar disorder. METHODS: We systematically searched the scientific literature up to September 2020 for experimental or observational studies which measured circadian rhythm in bipolar patients taking lithium (in comparison with placebo or other active treatments) and carried out a meta-analysis. Circadian rest-activity was our primary outcome, but we also collected data about sleep quality and chronotype (Morningness-Eveningness). The protocol was registered in PROSPERO (CRD42018109790). RESULTS: Four observational studies (n = 668) and one experimental study (n = 29) were included. Results from the meta-analysis suggest a potential association between lithium and shifts towards morningness (standardized mean difference [SMD]: 0.42, 95% confidence interval [CI]: -0.05 to 0.90). One cohort study with 21 days of follow-up found that patients treated with lithium had significantly larger amplitude (0.68, 0.01 to 1.36) when compared to anticonvulsants. CONCLUSION: This review highlights the insufficient evidence to inform us about the effect of lithium on circadian rhythm. However, we found that chronotype can be a potential target for further exploration of biomarkers or biosignatures of lithium treatment in patients with bipolar disorder. Further studies with prospective and longitudinal study design, adopting actigraphy to monitor daily circadian rest-activity changes are needed.

Effect evaluation of “healthy eating plate” based dietary management for diabetic inpatients / 预防医学

Objective@#To evaluate the effect of "healthy eating plate" based dietary management on diabetic inpatients.@*Methods@#The patients with type 2 diabetes mellitus (T2DM) admitted to Daishan First People's Hospital from November 2019 to November 2020 were selected and randomly divided into two groups. The intervention group was given "healthy eating plate" based dietary management, while the control group was given routine dietary management. Demographic data and physical examination results were collected. Fasting blood glucose (FPG), glycosylated hemoglobin (HbA1c), triglyceride (TG) and total cholesterol (TC) were detected at admission, discharge and 3 months after discharge, and compared between the two groups by covariance and generalized estimating equation. @*Results@#here were 52 patients aged (55.83±9.67) years in the intervention group, with 29 (55.77%) males and 23 (44.23%) females. There were 53 patients aged (57.54±11.09) years in the control group, with 32 (60.38%) males and 21 (39.62%) females. There were no significant differences in FPG, HbA1c, TG and TC levels between two groups at discharge (P>0.05). The level of HbA1c in the intervention group was significantly lower than that in the control group at 3 months after discharge (P<0.05); there were no significant differences in FPG, TG and TC levels (P>0.05).@*Conclusion @#The "healthy eating plate" based dietary management can better control the blood glucose of diabetic patients, and can help maintain the dietary treatment. It is worthy of promotion in diabetic patients.

Association between bone marrow fluorodeoxyglucose uptake and recurrence after curative surgical resection in patients with T1-2N0M0 lung adenocarcinoma: a retrospective cohort study.

BACKGROUND: Evidence regarding the relationship between fluorodeoxyglucose (FDG) uptake in the bone marrow of patients with lung adenocarcinoma and prognosis is limited. This study aimed to identify whether bone marrow FDG uptake is a risk factor for recurrence in patients after curative surgical resection of T1-2N0M0 lung adenocarcinoma. METHODS: From January 2012 to December 2016, we retrospectively enrolled 195 pT1-2N0M0 lung adenocarcinoma patients who underwent both preoperative FDG positron emission tomography/computed tomography (PET/CT) and surgical resection from the lung adenocarcinoma database maintained by the PET/CT department at our hospital. After surgical resection, patients were followed up mainly through regular outpatient examinations. The maximum standardized uptake value (SUVmax) of the primary tumor, the mean FDG uptake of bone marrow (BM SUV), bone marrow-liver uptake ratio (BLR), metabolic tumor volume (MTV), and total lesion glycolysis (TLG) were measured from the pretreatment FDG PET/CT images. Multi-adjusted Cox proportional hazards models were built to evaluate the independent prognostic value of BLR in predicting recurrence-free survival (RFS). A restricted cubic spline regression model was conducted to provide more precise estimates and examine the shape of the associations between BLR and the risk of recurrence. RESULTS: The follow-up results showed that 30 of the 195 patients (15.4%) had tumor recurrence. Compared with non-recurrent patients, the primary tumor size in recurrent patients was larger, and the SUVmax, TLG, and serum C-reactive protein (CRP) levels were higher. Univariate analysis showed that BLR, tumor size, SUVmax, TLG, and CRP were significantly correlated with postoperative tumor recurrence. After adjustment for conventional confounding factors, the hazard ratio of BLR was 5.01 (95% CI, 1.32, 18.98) for the highest tertile of BLR compared with the lowest tertile. The multi-adjusted spline regression showed that BLR had a linear relationship with log relative risk (RR) for recurrence when BLR was lower than 0.7. Over this level, the effect stabilized, suggesting a saturation effect for BLR at a level of approximately 0.7 at recurrence. CONCLUSIONS: BLR was an independent risk factor for predicting RFS in T1-2N0M0 lung adenocarcinoma patients after curative surgical resection. BLR can be used as a biomarker for evaluating the risk of lung cancer recurrence.

Reliability of Atmosphere Pressure-Plasma Enhanced Chemical Vapor Deposition Deposited Indium Gallium Zinc Oxide Resistive Random Access Memory Device with Microwave Annealing.

Recently resistive random access memory (RRAM) is considered to be the most promising one to become the next generation memory since its simple Metal/Insulator/Metal (MIM) structure, lower power consumption and fabrication cost (Meena, J.S., et al., 2014. Overview of emerging nonvolatile memory technologies. Nanoscale Research Letters, 9(1), p.526). Due to some bottlenecks for current flash memory, such as high operation voltage, low operation speed, poor retention time and endurance, RRAM device is regarded as an alternative solution (Fuh, C.S., et al., 2011. Role of environmental and annealing conditions on the passivation-free In-Ga-Zn-O TFT. Thin Solid Films, 520, pp.1489-1494). In this investigation, the memory layer of RRAM device is IGZO, and it is deposited with AP-PECVD technique which can operate under atmosphere, reduce cost of the process. Microwave annealing (MWA) is used to enhance the RRAM device reliability (Fuh, C.S., et al., 2011. Role of environmental and annealing conditions on the passivation-free In-Ga-Zn-O TFT. Thin Solid Films, 520, pp.1489-1494). Experiment shows that with appropriate MWA treatment, the IGZO RRAM device exhibits better electrical characteristics, reliability issues such as numbers of switching cycle and data retention time are also improved (Teng, L.F., et al., 2012. Effects of microwave annealing on electrical enhancement of amorphous oxide semiconductor thin film transistor. Applied Physics Letters, 101, p.132901).

Effects of P-Type SnOx Thin-Film Transistors with N2 and O2 Ambient Furnace Annealing.

Recently oxide-based thin-film transistors (TFTs) are investigated for emerging applications of the next generation display devices and other electronic circuits (Fortunato, E., et al., 2012. Oxide semiconductor thin-film transistors: A review of recent advances. Advanced Materials, 24, pp.2945-2986). Despite of the great success in n-type oxide semiconductors with high transparency and high field-effect mobility, high performance P-type oxide TFTs are so highly desired that complementary circuits can be realized with low power and high performance (Ou, W.C., et al., 2008. Anomalous P-channel amorphous oxide transistors based on tin oxide and their complementary circuits. Applied Physics Letters, 92, p.122113). There are some oxides such as SnO, CuO, Cu2O and NiO are regarded as promising P-type semiconductor materials. In this investigation, tin oxide SnOx is fabricated to be active layer for TFTs device, and furnace annealing with several combinations of nitrogen and oxygen ambient is compared to enhance the electrical characteristics of P-type SnOx TFTs (Park, K.S., et al., 2009. High performance solution-processed and lithographically patterned zinc-tin oxide thin-film transistors with good operational stability. Electrochemical and Solid-State Lett., 12, pp.H256-H258). The results show that with N2+O2 ambient, 30 minutes furnace annealing, the P-type SnOx TFTs device shows better performance with mobility (µFE) 0.883 cm²/V · S, threshold voltage (VT) -4.63 V, subthreshold swing (SS) 1.15 V/decade, and Ion/Ioff ratio 1.01×103.

Study of Atmospheric-Pressure Plasma Enhanced Chemical Vapor Deposition Fabricated Indium Gallium Zinc Oxide Thin Film Transistors with In-Situ Hydrogen Plasma Treatment.

Amorphous InGaZnO (a-IGZO) Thin Film Transistors (TFTs) has been studied extensively for their perspective applications in next generation active-matrix displays such as liquid crystal displays and flat-panel displays, due to its better field-effect mobility (>10 cm²/V · S), larger Ion/Ioff ratio (>106), and better stability electrical. Hydrogen is known as shallow donors for n-type (channel) oxide semiconductors (Dong, J.J., et al. 2010. Effects of hydrogen plasma treatment on the electrical and optical properties of Zno films: Identification of hydrogen donors in ZnO. ACS Appl. Mater. Interfaces, 2, pp.1780-1784), and it is also effective passivator for traps (Tsao, S.W., et al., 2010. Hydrogen-induced improvements in electrical characteristics of a-IGZO thin-film transistors. Solid-State Electron, 54, pp.1497-1499). In this study, In-Situ hydrogen plasma is applied to deposit IGZO channel. With atmospheric-pressure PECVD (AP-PECVD), IGZO thin film can be deposited without vacuum system, large area manufacturing, and cost reducing (Chang, K.M., et al., 2011. Transparent conductive indium-doped zinc oxide films prepared by atmospheric pressure plasma jet. Thin Solid Films, 519, pp.5114-5117). The results show that with appropriate flow ratio of Ar/H2 plasma treatment, the a-IGZO TFT device exhibits better performance with mobility (µFE) 19.7 cm²/V · S, threshold voltage (VT) 1.18 V, subthreshold swing (SS) 81 mV/decade, and Ion/Ioff ratio 5.35×107.

Investigation of Microwave Annealing on Resistive Random Access Memory Device with Atmospheric Pressure Plasma Enhanced Chemical Vapor Deposition Deposited IGZO Layer.

Non-volatile memory (NVM) is essential in almost every consumer electronic products. The most prevalent NVM used nowadays is flash memory (Meena, J.S., et al., 2014. Overview of emerging nonvolatile memory technologies. Nanoscale Res. Letters, 9(1), p.526). However, some bottlenecks of flash memory have been identified, such as high operation voltage, low operation speed, and poor retention time. Resistive random access memory (RRAM) is considered to be the most promising one to become the next generation NVM device since its simple structure, fast program/erase speed, and low power consumption. In this experiment, the RRAM device is fabricated, and its IGZO (memory) layer is deposited with AP-PECVD technique which can reduce cost of the process. Microwave annealing (MWA) is used to enhance electrical characteristics of the RRAM device (Fuh, C.S., et al., 2011. Role of environmental and annealing conditions on the passivation-free In-Ga- Zn-O TFT. Thin Solid Films, 520, pp.1489-1494). Experiment results show that with appropriate MWA treatment, the IGZO RRAM device exhibits better electrical characteristics under bipolar operation, all forming/set/reset voltage for RRAM device is simultaneously lowered.

Cognitive Impairment in Patients with Bipolar Disorder: Impact of Pharmacological Treatment.

Bipolar disorder is an illness characterised by periods of elated and depressed mood. These mood episodes are associated with changes in cognitive function and there is evidence to suggest that cognitive dysfunction persists during euthymia. The extent to which this is a function of the illness or a result of treatment is less clear. In this narrative review, we explore the impact of commonly used medications for bipolar disorder on cognitive function. Specific impairments in executive function and verbal memory have been noted in bipolar disorder. The impact of pharmacological treatments upon cognitive function is mixed with a number of studies reporting conflicting results. Interpretation of the data is further complicated by the variety of cognitive tests employed, study design, the relatively small numbers of patients included and confounding by indication. Overall, there is some evidence that while lithium improves some cognitive domains, it impedes others. Antipsychotics may be deleterious to cognition, although this may relate to the patient population in which they are prescribed. Sodium valproate is also associated with worse cognitive outcomes, while the impact of other antiepileptics is unclear. Overall the quality of evidence is poor and is derived from a relatively small number of studies that often do not account for the significant heterogeneity of the disorder or common comorbidities. The use of consistent methodologies and measures of cognition across studies, as well as in naturalistic settings, would enable more certain conclusions to be drawn.

Noncanonical ATG8-ABS3 interaction controls senescence in plants.

Protein homeostasis is essential for cellular functions and longevity, and the loss of proteostasis is one of the hallmarks of senescence. Autophagy is an evolutionarily conserved cellular degradation pathway that is critical for the maintenance of proteostasis. Paradoxically, autophagy deficiency leads to accelerated protein loss by unknown mechanisms. We discover that the ABNORMAL SHOOT3 (ABS3) subfamily of multidrug and toxic compound extrusion transporters promote senescence under natural and carbon-deprivation conditions in Arabidopsis thaliana. The senescence-promoting ABS3 pathway functions in parallel with the longevity-promoting autophagy to balance plant senescence and survival. Surprisingly, ABS3 subfamily multidrug and toxic compound extrusion proteins interact with AUTOPHAGY-RELATED PROTEIN 8 (ATG8) at the late endosome to promote senescence and protein degradation without canonical cleavage and lipidation of ATG8. This non-autophagic ATG8-ABS3 interaction paradigm is probably conserved among dicots and monocots. Our findings uncover a previously unknown non-autophagic function of ATG8 and an unrecognized senescence regulatory pathway controlled by ATG8-ABS3-mediated proteostasis.

Balance between Cytosolic and Chloroplast Translation Affects Leaf Variegation.

The development of functional chloroplasts relies on the fine coordination of expressions of both nuclear and chloroplast genomes. We have been using the Arabidopsis (Arabidopsis thaliana) yellow variegated (var2) leaf variegation mutant as a tool to dissect the regulation of chloroplast development. In this work, we screened for var2 genetic enhancer modifiers termed enhancer of variegation (evr) mutants and report the characterization of the first EVR locus, EVR1 We showed that EVR1 encodes the cytosolic 80S ribosome 40S small subunit protein RPS21B and the loss of EVR1 causes the enhancement of var2 leaf variegation. We further demonstrated that combined S21 activities from EVR1 and its close homolog, EVR1L1, are essential for Arabidopsis, and they act redundantly in regulating leaf development and var2 leaf variegation. Moreover, using additional cytosolic ribosomal protein mutants, we showed that although mutations in cytosolic ribosomal proteins all enhance var2 leaf variegation to varying degrees, the 40S subunit appears to have a more profound role over the 60S subunit in regulating VAR2-mediated chloroplast development. Comprehensive genetic analyses with var2 suppressors that are defective in chloroplast translation established that the enhancement of var2 leaf variegation by cytosolic ribosomal protein mutants is dependent on chloroplast translation. Based on our data, we propose a model that incorporates the suppression and enhancement of var2 leaf variegation, and hypothesize that VAR2/AtFtsH2 may be intimately involved in the balancing of cytosolic and chloroplast translation programs during chloroplast biogenesis.

WM130 preferentially inhibits hepatic cancer stem-like cells by suppressing AKT/GSK3β/β-catenin signaling pathway / 中国药理学与毒理学杂志

OBJECTIVE The eradication of cancer stem cells(CSCs)is signifcant for cancer therapy and prevention.METHODS In this study,we evaluated WM130,a novel derivative of matrine,for its effect on CSCs using human hepatocellular carcinoma(HCC)cell lines,their sphere cells,and sorted EpCAM+cells. RESULTS We revealed that WM130 could not only inhibit proliferation and colony formation of HCC cells, but also suppress the expression of some stemness-related genes and up-regulate some mature hepatocyte marker genes, indicating a promotion of differentiation from CSCs to hepatocytes. WM130 also suppressed the proliferation of doxorubicin-resistant hepatoma cells, and markedly reduced the cells with CSC biomarker EpCAM.Moreover,WM130 suppressed HCC spheres,not only primary spheres but also subsequent spheres,indicating an inhibitory effect on self-renewal capability of CSCs.Interestingly,WM130 exhibiteda remarkable inhibitory preference on HCC spheres and EpCAM+cells rather than their parental HCC cells and EpCAM- cells respectively. In vivo, WM130 inhibited HCC xenograft growth, decreased the number of sphere-forming cells, and remarkably decreased the levels of EpCAM mRNA and protein in tumor xenografts. Better inhibitory effect was achieved by WM130 in combination with doxorubicin.Further mechanism study revealed that WM130 inhibited AKT/GSK3β/β-catenin signaling pathway. CONCLUSION Collectively, our results suggest that WM130 remark-ably inhibits hepatic CSCs, and this effect may via the down-regulation of the AKT/GSK3β/β-catenin pathway.These findings provide a strong rationale for the use of WM130 as a novel drug candidate in HCC therapy.

In situ functional dissection of RNA cis-regulatory elements by multiplex CRISPR-Cas9 genome engineering.

RNA regulatory elements (RREs) are an important yet relatively under-explored facet of gene regulation. Deciphering the prevalence and functional impact of this post-transcriptional control layer requires technologies for disrupting RREs without perturbing cellular homeostasis. Here we describe genome-engineering based evaluation of RNA regulatory element activity (GenERA), a clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 platform for in situ high-content functional analysis of RREs. We use GenERA to survey the entire regulatory landscape of a 3'UTR, and apply it in a multiplex fashion to analyse combinatorial interactions between sets of miRNA response elements (MREs), providing strong evidence for cooperative activity. We also employ this technology to probe the functionality of an entire MRE network under cellular homeostasis, and show that high-resolution analysis of the GenERA dataset can be used to extract functional features of MREs. This study provides a genome editing-based multiplex strategy for direct functional interrogation of RNA cis-regulatory elements in a native cellular environment.

Sphere-Induced Rejuvenation of Swine and Human Müller Glia Is Primarily Caused by Telomere Elongation.

Müller cells are the major supportive and protective glial cells in the retina with important functions in histogenesis and synaptogenesis during development, and in maintenance of mature neurons as they show to secrete various cytokines and manifest potentials of self-renewal and transdifferentiation into retinal neurons following injury in the vertebrate retinas. The swine retina has a visual streak structure similar to the human macular where cone photoreceptors are highly concentrated, thereby can serve as a better model for studying retinal diseases and for formulating cell-based therapeutics than the rodent retinas. Like most differentiated somatic mammalian cells, the isolated swine and human Müller glia become senescent over passages in culture, which restricts their potential application in basic and clinic researches. Here, we demonstrate that the senescence of swine and human Müller cells is caused by telomere attrition upon multiplications in vitro; and the senescent cells can be rejuvenated by sphere suspension culture. We also provide evidence that sphere-induced extension of telomeres in swine and human Müller glia is achieved by alternative lengthening of telomeres or/and by telomerase activation. Stem Cells 2017;35:1579-1591.

Inhibition of YAP/TAZ Activity in Spinal Cord Suppresses Neuropathic Pain.

UNLABELLED: Neuropathic pain, often caused by nerve injury, is a major clinical challenge. Mechanisms that underlie neuropathic pain remain elusive and effective medications are limited. Numerous investigations of pain mechanisms have focused on alterations and phenotypic switches of the nociceptive transmitters and modulators, as well as on their receptors and downstream signaling pathways that have already exerted roles in the pain processes of mature nervous systems. We have demonstrated recently that nerve injury may elicit neuronal alterations that recapitulate events occurring during development. Signaling of the representative activated molecule Wnt thus becomes a trigger for the development of neuropathic pain and is a potential therapeutic target. We report that the transcriptional regulators YAP and TAZ, which orchestrate Wnt response via incorporation in the ß-catenin destruction complex, are key in the pathogenesis of neuropathic pain and may serve as an "ON-OFF" switch for neuropathic pain status in rats. Peripheral nerve injury causes rapid-onset and long-lasting nuclear accumulation of YAP/TAZ/ß-catenin in the spinal dorsal horn. Spinal inhibition or knock-down of either YAP or TAZ suppresses mechanical allodynia induced by nerve injury or the pain initiators lysophosphatidic acid and Wnt3a. Promoting the nuclear accumulation of YAP/TAZ leads to mechanical hypersensitivity in naive animals. Further, we discovered a new small molecule, dCTB, which targets YAP/TAZ/ß-catenin and can greatly suppress neuropathic pain and the associated neurochemical alterations. Our study reveals that YAP and TAZ are core mechanisms underlying the pathogenesis of neuropathic pain and are targets in the screening for potent analgesics for the treatment of neuropathic pain. SIGNIFICANCE STATEMENT: Mechanisms that underlie neuropathic pain remain elusive. We have demonstrated recently that nerve injury can activate Wnt signaling, which becomes a trigger for the development of neuropathic pain. We report that the transcriptional regulators YAP and TAZ, which orchestrate Wnt response via incorporation in the ß-catenin destruction complex, are key in the pathogenesis of neuropathic pain and may serve as an "ON-OFF" switch for neuropathic pain status. Further, we discovered a new small molecule, dCTB, which targets YAP/TAZ/ß-catenin and can greatly suppress neuropathic pain. Our study reveals that YAP and TAZ are core mechanisms underlying the pathogenesis of neuropathic pain and are targets in the screening of potent analgesics for the treatment of neuropathic pain.