Circulating exosomal circRNA-miRNA-mRNA network in a familial partial lipodystrophy type 3 family with a novel PPARG frameshift mutation c.418dup.

Familial Partial Lipodystrophy 3 (FPLD3) is a rare genetic disorder caused by loss-of-function mutations in the PPARG gene, characterized by a selective absence of subcutaneous fat and associated metabolic complications. However, the molecular mechanisms of FPLD3 remain unclear. In this study, we recruited a 17-year-old Chinese female with FPLD3 and her family, identifying a novel PPARG frameshift mutation (exon 4: c.418dup: p.R140Kfs*7) that truncates the PPARγ protein at the 7th amino acid, significantly expanding the genetic landscape of FPLD3. By performing next generation sequencing of circular RNAs (circRNAs), microRNAs (miRNAs), and mRNAs in plasma exosomes, we discovered 59 circRNAs, 57 miRNAs, and 299 mRNAs were significantly altered in the mutation carriers in the comparison of healthy controls. Integration analysis highlighted that the circ_0001597-miR-671-5p pair and 18 mRNAs might be incorporated into the metabolic regulatory networks of the FPLD3 induced by the novel PPARG mutation. Functional annotation suggested that these genes were significantly enriched in glucose and lipid metabolism related pathways. Among the circRNA-miRNA-mRNA network, we identified two critical regulators, EGR1, a key transcription factor known for its role in insulin signaling pathways and lipid metabolism, and AGPAT3, which gets involved in the biosynthesis of triglycerides and lipolysis. Circ_0001597 regulates the expression of these genes through miR-671-5p, potentially contributing to the pathophysiology of FPLD3. Overall, this study clarified a circulating exosomal circRNA-miRNA-mRNA network in a FPLD3 family with a novel PPARG mutation, providing evidence for exploring promising biomarkers and developing novel therapeutic strategies for this rare genetic disorder.

Quercetin relieved diabetic neuropathic pain by inhibiting upregulated P2X4 receptor in dorsal root ganglia.

The upregulation of nociceptive ion channels expressed in dorsal root ganglia (DRG) contributes to the development and retaining of diabetic pain symptoms. The flavonoid quercetin (3,3',4',5,7-pentahydroxyflavone) is a component extracted from various fruits and vegetables and exerts anti-inflammatory, analgesic, anticarcinogenic, antiulcer, and antihypertensive effects. However, the exact mechanism underlying quercetin's analgesic action remains poorly understood. The aim of this study was to investigate the effects of quercetin on diabetic neuropathic pain related to the P2X4 receptor in the DRG of type 2 diabetic rat model. Our data showed that both mechanical withdrawal threshold and thermal withdrawal latency in diabetic rats treated with quercetin were higher compared with those in untreated diabetic rats. The expression levels of P2X4 messenger RNA and protein in the DRG of diabetic rats were increased compared with the control rats, while quercetin treatment significantly inhibited such enhanced P2X4 expression in diabetic rats. The satellite glial cells (SGCs) enwrap the neuronal soma in the DRG. Quercetin treatment also lowered the elevated coexpression of P2X4 and glial fibrillary acidic protein (a marker of SGCs) and decreased the upregulation of phosphorylated p38 mitogen-activated protein kinase (p38MAPK) in the DRG of diabetic rats. Quercetin significantly reduced the P2X4 agonist adenosine triphosphate-activated currents in HEK293 cells transfected with P2X4 receptors. Thus, our data demonstrate that quercetin may decrease the upregulation of the P2X4 receptor in DRG SGCs, and consequently inhibit P2X4 receptor-mediated p38MAPK activation to relieve the mechanical and thermal hyperalgesia in diabetic rats.

Anxiety screening tools in people with epilepsy: A systematic review of validated tools.

OBJECTIVE: Anxiety is a common neurological condition often comorbid with epilepsy, with approximately 20% of patients with epilepsy exhibiting symptoms of anxiety. Despite this prevalence, accurate and efficacious tools designed to screen for anxiety specifically in patients with epilepsy have not yet been developed. The purpose of this study is to systematically review the literature and better understand this relationship. METHODS: Ovid MEDLINE, EMBASE, and PsyclNFO were searched until April 22nd, 2019 without language restrictions. We extracted abstracts, data abstraction, and full-text reviews in duplicate and chose the studies that included measures for anxiety screening in patients with epilepsy. The Quality Assessment of Diagnostic Accuracy Studies Version was used to assess study quality. We used the medians and ranges to calculate the accuracy of the tools. RESULTS: We screened 4758 abstracts and selected 11 articles dealing with anxiety. The most common validated anxiety screening tools were the Generalized Anxiety Disorder (GAD-7) and Hospital Anxiety and Depression Scale-A (HADS-A). The Mini International Neuropsychiatric Interview (MINI) was the most common reference standard used. SIGNIFICANCE: Many studies have validated depression screening tools rather than anxiety. This lack of data has left much uncertainty about the relationship of epilepsy to anxiety, as well as diagnostic inconsistencies. The effectiveness of these assessments in practice may be overestimating the prevalence, as the cutpoints are usually chosen after the fact, based on the study sample.

Efficacy Comparison of Six Chemotherapeutic Combinations for Osteosarcoma and Ewing's Sarcoma Treatment: A Network Meta-Analysis.

This study aimed to address the insufficiency of traditional meta-analysis and provide improved guidelines for the clinical practice of osteosarcoma treatment. The heterogeneity of the fixed-effect model was calculated, and when necessary, a random-effect model was adopted. Furthermore, the direct and indirect evidence was pooled together and exhibited in the forest plot and slash table. The surface under the cumulative ranking curve (SUCRA) value was also measured to rank each intervention. Finally, heat plot was introduced to demonstrate the contribution of each intervention and the inconsistency between direct and indirect comparisons. This network meta-analysis included 32 trials, involving a total of 5,626 subjects reported by 28 articles. All the treatments were classified into six chemotherapeutic combinations: dual agent with or without ifosfamide (IFO), multi-agent with or without IFO, and dual agent or multi-agent with IFO and etoposide. For the primary outcomes, both overall survival (OS) and event-free survival (EFS) rates were considered. The multi-agent integrated with IFO and etoposide showed an optimal performance for 5-year OS, 10-year OS, 3-year EFS, 5-year EFS, and 10-year EFS when compared with placebo. The SUCRA value of this treatment was also the highest of these six interventions. However, multi-drug with IFO alone had the highest SUCRA value of 0.652 and 0.516 when it came to relapse and lung-metastasis. It was efficient to some extent, but no significant difference was observed in both outcomes. Chemotherapy, applied as induction or adjuvant treatment with radiation therapy or surgery, is able to increase the survival rate of patients, especially by combining multi-drug with IFO and etoposide, which demonstrated the best performance in both OS and EFS. As for relapse and the lung-metastasis, multiple agents with IFO alone seemed to have the optimal efficiency, although no significant difference was observed here. J. Cell. Biochem. 119: 250-259, 2018. © 2017 Wiley Periodicals, Inc.

Effects of er-miao-san extracts on TNF-alpha-induced MMP-1 expression in human dermal fibroblasts.

BACKGROUND: Various health benefits have been attributed to Er-Miao-San (EMS), a traditional Chinese herbal formulation that contains equal amounts of cortex phellodendri (Phellodendron amurense Ruprecht) and rhizoma atractylodis (Atractylodes lancea D.C). However, its effect on the anti-inflammatory activity in human dermal fibroblasts (HDFs) and the mechanism underlying this effect are unknown. RESULTS: This study investigated the effects of EMS on TNF-α-induced MMP-1 expression in HDFs. Our data show that EMS inhibited TNF-α-induced MMP-1 expression in a concentration-dependent manner. Interestingly, EMS maintained IκB content without inhibiting the phosphorylation of MAPKs, which are well-established upstream kinases of NF-κB. Moreover, EMS reduced the level of nuclear p65 protein in HDFs. Luciferase assay revealed that EMS inhibits the transcriptional activity of NF-κB by stabilizing IκB. Our results show that EMS exerts its anti-inflammatory effect by inhibiting NF-κB-regulated genes such as IL-1ß and IL-8. Moreover, EMS effectively inhibited TNF-α-induced expression of MMP-1 via the NF-κB pathway. CONCLUSIONS: Taken together, our data suggest that EMS could potentially be used as an anti-inflammatory and anti-aging treatment.

Combination of Pioglitazone and Metformin Actions on Liver Lipid Metabolism in Obese Mice.

BACKGROUND: Despite the increasing prevalence rate of nonalcoholic fatty liver disease (NAFLD) worldwide, efficient pharmacotherapeutic regimens against NAFLD still need to be explored. Previous studies found that pioglitazone and metformin therapy could partly ameliorate NAFLD, but their combination therapy effects have not been researched. In the present study, we assessed the protective effects of metformin and pioglitazone combination therapy on liver lipid metabolism in high-fat diet (HFD)-fed mice and investigated the molecular mechanism. METHODS: Male C57BL/6 mice were divided into five groups: normal control; HFD control; metformin monotherapy; pioglitazone monotherapy and combined therapy. After 8 weeks of pharmacological intervention, glucose and lipid metabolism characteristics, hepatic histology, lipidomics profiling and RNA-seq analysis were performed. RESULTS: The combination of pioglitazone and metformin significantly ameliorated HFD-induced metabolic disturbance and the hepatic oil red O area. A lipidomics analysis showed that combined therapy could significantly reduce the high levels of free fatty acids (FFA), diacylglycerol and triglycerides, while a set of glycerophospholipids and sphingolipids were increased in the combined therapy group. Consistently, an RNA-seq analysis also showed a remarkable reduction in genes associated with FFA uptake and de novo lipogenesis, including Cd36, Fads1, Fads2, Fasn, Scd1, Elovl5 and Pklr in the combined therapy group. CONCLUSIONS: Pioglitazone and metformin might have a synergistic protective effect on NAFLD by improving hepatic lipid profiles in HFD-induced mice. Further studies are needed to verify the clinical effects.

Turn-on luminescent sensing of metal cations via quencher displacement: rational design of a highly selective chemosensor for chromium(III).

A highly selective luminescent chemosensor for Cr(3+) in aqueous solution was assembled by a low-selectivity luminogenic receptor with Cu(2+) as a metal quencher. Three tetranitrogen chelating sites were integrated into the multichannel receptor with a tris(1,10-phenanthroline)ruthenium(II) luminophore at the core. This receptor (2) exhibits chelating affinity for many transition-metal cations, among which Cu(2+) efficiently quenches the emission. The further addition of Cr(3+) into the Cu(2+)-titrated solution of 2 results in a metal-exchange reaction and a sensitive turn-on luminescence response highly selective over other metal cations. The quencher displacement sensing strategy in this design can be a simple but efficient approach for OFF-ON luminescent sensing of metal cations that inherently lack selective ligands.

Maternal Exercise Programs Glucose and Lipid Metabolism and Modulates Hepatic miRNAs in Adult Male Offspring.

Detrimental exposures in mothers are recognized as risk factors for the development of metabolic dysfunction in offspring. In contrast, maternal exercise has been reported to be an effective strategy to maintain offspring health. However, the mechanisms underlying the protective effects of maternal exercise on adult offspring metabolic homeostasis are largely unclear. This study aims to investigate whether maternal exercise before and during pregnancy could combat the adverse effects of maternal high-fat diet (HFD) on metabolism in 24-week-old male offspring and to explore the role of miRNAs in mediating the effects. Female C57BL/6 mice were fed with either control diet or HFD 3-week prior to breeding and throughout pregnancy and lactation, among whom half of the HFD-fed mice were submitted to voluntary wheel running training 3-week before and during pregnancy. Male offspring were sedentary and fed with a control diet from weaning to 24 weeks. Body weight, the content of inguinal subcutaneous adipose tissue and perirenal visceral adipose tissue, glucose tolerance, and serum insulin and lipids in offspring were analyzed. Hepatic tissues were collected for transcriptome and miRNA sequencing and reverse transcription-quantitative polymerase chain reaction validation. The results showed that maternal HFD resulted in significant glucose intolerance, insulin resistance, and dyslipidemia in adult offspring, which were negated by maternal exercise. Transcriptome sequencing showed that maternal exercise reversed perinatal HFD-regulated genes in adult offspring, which were enriched in glucose and lipid metabolic-related signaling pathways. At the same time, maternal exercise significantly rescued the changes in the expression levels of 3 hepatic miRNAs in adult offspring, and their target genes were involved in the regulation of cholesterol biosynthesis and epigenetic modification, which may play an important role in mediating the intergenerational metabolic regulation of exercise. Overall, our research pioneered the role of miRNAs in mediating the programming effects of maternal exercise on adult offspring metabolism, which might provide novel insight into the prevention and treatment of metabolic disorders in early life.

Genistein improves glucose metabolism and promotes adipose tissue browning through modulating gut microbiota in mice.

Genistein has beneficial effects on glucose metabolism and adipose tissue browning which paralleled with gut microbiota alterations. However, the causality is unclear. This study aims to evaluate whether genistein could ameliorate metabolism and activate the browning program and whether gut microbiota is indispensable for these alterations. We examined the effect of 10-week genistein gavage (30 mg kg-1 d-1) on glucose metabolism in C57BL/6J mice. Cecal contents were collected for 16s rRNA sequencing. The mRNA of browning markers was quantified in adipose tissues. Antibiotic administration was used to deplete gut microbiota. The lean mice with a normal control diet and genistein exhibited better glucose tolerance and higher expression of UCP1 and PGC1α in white fat compared with those without genistein. Markedly enriched Blautia, Ruminiclostridium_5, and Ruminiclostridium_9 in genistein-treated mice were significantly correlated with browning markers and glucose tolerance. In obese mice, genistein alleviated the detrimental effects of a high-fat diet on glucose homeostasis and increased UCP1 and PGC1α expression in brown fat. Obvious increases in Ruminiclostridium, Rikenella, and Clostridium_sensu_stricto_1 by genistein were associated with metabolic improvement. However, depleting gut microbiota abolished these benefits. Overall, our findings indicated that gut microbiota contributed to enhanced glucose metabolism and adipose tissue browning of genistein, providing a promising target for metabolic health protection.

A high-fat diet disrupts the hepatic and adipose circadian rhythms and modulates the diurnal rhythm of gut microbiota-derived short-chain fatty acids in gestational mice.

The prevalence of gestational obesity has reached epidemic proportions. Evidence supported that the interactions between the gut microbiota and circadian clocks far reached, affecting host metabolism. Our study aimed to investigate the effect of a high-fat diet (HF) on the hepatic and adipose circadian rhythms in gestational mice and to explore the role of gut microbiota-derived short-chain fatty acids (SCFAs) in mediating the effects. C57BL/6 female mice were randomly fed a standard chow diet (Ctr) or HF prior to and during pregnancy. Samples were collected every 4 h over 24 h (six time points), and 16S rRNA and metabonomics were carried out. Rhythmic patterns were identified and compared using CircaCompare. The results showed that the HF before and during pregnancy significantly induced obesity and worsen glucose tolerance, insulin sensitivity, and lipid metabolism in the gestational mice. Furthermore, the HF significantly disrupted the rhythmic pattern of hepatic and adipose circadian clock genes and downstream metabolic genes. Importantly, our results revealed that the HF altered the diurnal rhythm of the gut microbiota in a diverse manner, which was assessed across three categories: phase shift, loss rhythmicity, and gained rhythmicity. We report here, for the first time, a parallel alteration of the rhythmic phase of butyric acid and butyrate-producing Clostridiaceae_1, which was confirmed by a positive correlation between them. Overall, our research emphasized the importance of the rhythmicity of gut microbiota-derived SCFAs in mediating circadian disruption in response to the HF in gestational mice, which may provide novel insights into the prevention and treatment of gestational obesity.

The effect of sodium-glucose cotransporter 2 inhibitors on biomarkers of inflammation: A systematic review and meta-analysis of randomized controlled trials.

Aims: Inflammatory biomarkers may play vital roles in the pathophysiology of diabetes and diabetic cardiorenal complications. Sodium-glucose cotransporter-2 (SGLT2) inhibitors have a potential cardiovascular and renal protective effect in type 2 diabetes. The aim of this meta-analysis was to quantify the effects of SGLT2 inhibitors on biomarkers of inflammation in randomized controlled trials (RCTs). Methods: PubMed, Cochrane Library, EMBASE, and Web of Science were searched for eligible RCTs of adults with type 2 diabetes (T2D) with no time limit (updated to 12 October 2022). The biomarkers selected included C-reactive protein (CRP), interleukin-6, tumor necrosis factor-alpha, leptin, adiponectin, ferritin, plasminogen activator inhibitor (PAI)-1, and vascular cell adhesion molecule-1. Data were analyzed using a random-effect model in Review Manager 5.4. Results: Thirty-four studies with 6,261 patients (68.6% male) were eligible for this meta-analysis. The mean age of the participants was 62.57(±11.13) years old, and the median treatment duration length with follow-up was 24 weeks. Generally, the included trials were of good methodological quality. The meta-analysis revealed that ferritin levels were significantly reduced in SGLT2 inhibitor treatment groups versus placebo or standard diabetes therapies (SMD: -1.21; 95% CI: -1.91, -0.52, p < 0.001). The effects of CRP (SMD: 0.25; 95% CI: -0.47, -0.03, p = 0.02) and leptin (SMD: -0.22; 95% CI: -0.43, -0.01, p = 0.04) were reduced, and the effects of adiponectin were improved (SMD: 0.28; 95% CI: 0.15, 0.41, p < 0.001) in placebo-controlled studies. PAI-1 levels were significantly reduced in studies controlled for diabetes therapies (SMD: -0.38; 95% CI: -0.61, -0.15, p = 0.001). Conclusion: This analysis provides strong evidence supporting anti-inflammatory effects of SGLT2 inhibitors in T2D subjects. The mechanisms and possible targets for the inflammation reducing and cardiorenal protective properties of SGLT2 inhibitors remain to be explored.

New highly stable metallabenzenes via nucleophilic aromatic substitution reaction.

Treatment of the ruthenabenzene [Ru{CHC(PPh(3))CHC(PPh(3))CH}Cl(2)(PPh(3))(2)]Cl (1) with excess 8-hydroxyquinoline in the presence of CH(3)COONa under air atmosphere produced the S(N)Ar product [(C(9) H(6)NO)Ru{CHC(PPh(3))CHC(PPh(3))C}(C(9)H(6)NO)(PPh(3))]Cl(2) (3). Ruthenabenzene 3 could be stable in the solution of weak alkali or weak acid. However, reaction of 3 with NaOH afforded a 7:1 mixture of ruthenabenzenes [(C(9)H(6)NO)Ru{CHC(PPh(3))CHCHC}(C(9)H(6)NO)(PPh(3))]Cl (4) and [(C(9)H(6)NO)Ru{CHCHCHC(PPh(3))C}(C(9)H(6)NO)(PPh(3))]Cl (5), presumably involving a P-C bond cleavage of the metallacycle. Complex 3 was also reactive to HCl, which results in a transformation of 3 to ruthenabenzene [Ru{CHC(PPh(3))CHC(PPh(3))C}Cl(2)(C(9)H(6)NO)(PPh(3))]Cl (6) in high yield. Thermal stability tests showed that ruthenabenzenes 4, 5, and 6 have remarkable thermal stability both in solid state and in solution under air atmosphere. Ruthenabenzenes 4 and 5 were found to be fluorescent in common solvents and have spectral behaviors comparable to those organic multicyclic compounds containing large π-extended systems.

pH-switchable inversion of the metal-centered chirality of metallabenzenes: opposite stereodynamics in reactions of ruthenabenzene with L- and D-cysteine.

We report herein the first study on the chemical interaction between metallabenzenes and bioactive molecules. Due to its unique stereoelectronic activities, a phenanthroline-derived ruthenabenzene [Ru{CHC(PPh(3))CHC(PPh(3))CH}Cl(C(12)H(8)N(2))(PPh(3))]Cl(2) (1) selectively binds cysteine in aqueous solution at physiological pH and then undergoes a dynamic inversion of configuration at the Ru center. The structure of the L-cysteine-binding product of 1 has been determined by means of X-ray diffraction. The replacement of the L-cysteine with the D form results in an inverted stereodynamic effect. Furthermore, the inversion process of the Ru-centered configuration could be conveniently controlled by a simple pH adjustment. This is attributed to the significant influence of a special intramolecular electrostatic interaction on the dynamic epimerization process of the cysteine-binding product.

Near-infrared chromogenic sensing of organotin species by a cyclopalladated azo dye.

A simple cyclopalladated complex of 4-(2-thiazolylazo)resorcinol showed a specific red-to-green colour change upon addition of organotin species in the acetonitrile-water medium.

Comparing the Efficacy and Safety of Low-Carbohydrate Diets with Low-Fat Diets for Type 2 Diabetes Mellitus Patients: A Systematic Review and Meta-Analysis of Randomized Clinical Trials.

INTRODUCTION: To compare the efficacy of low-carbohydrate diets (LCDs) with low-fat diets (LFDs) in body weight and glycemic control for type 2 diabetes mellitus (T2DM) patients, and their cardiovascular and renal safety. METHODS: We searched PubMed, Ovid, Embase databases, Cochrane Central Register of Controlled Trials (CENTRAL), and ClinicalTrials.gov from inception to April, 2021. Randomized controlled trials (RCTs) which lasted more than 3 months were included. The primary outcomes are the mean change from baseline in glycated haemoglobin (HbA1c) and body weight loss. Secondary outcomes included mean difference in lipid parameters, blood pressures, and serum creatinine. RESULTS: Totally, 12 RCTs met inclusion criteria representing 761 patients. Compared with LFDs, treatment with LCDs achieved significant reduced HbA1c by 0.35% (95% CI: -0.45, -0.24; P < 0.00001). LCDs appeared to be more beneficial in decreasing body weight than LFDs (WMD = -2.99 kg; 95% CI: -4.36, -1.63; P < 0.0001), especially in the subgroup that used VLCDs (WMD = -9.49 kg; 95% CI: -12.88, -6.09, P < 0.00001). For cardiovascular risk factors, the LCD interventions significantly reduced TG concentration (WMD: -0.20 mmol/l; 95% CI: -0.31, -0.10; P = 0.0001) and increased HDL-C concentration (WMD: 0.09 mmol/l; 95% CI: 0.05,0.13; P < 0.00001). Subgroup analyses demonstrated that the difference in HbA1c, TG, and HDL-C between two dietary restrictions respectively lasted up to 1.5 and 2 years, whereas the beneficial effects of body weight loss diminished over time and disappeared after 2 years. LCDs were not associated with decreased level of TC or LDL-C, neither SBP nor DBP in comparison with LFDs. Moreover, no significant difference in serum creatinine could be found among such two diet interventions. CONCLUSIONS: LCDs are superior to LFDs for T2DM patients in improving HbA1c and reducing body weight, with a rewarding effect of some cardiovascular risk factors in a longer-term diabetes management. However, available data are insufficient to evaluate the association between diet interventions and renal safety. Future larger longer-term follow-up clinical trials are needed to provide more evidence about the sustainable effects and safety of LCDs compared with LFDs.

Compounds of traditional Chinese medicine and neuropathic pain.

Neuropathic pain (NP) has become a serious global health issue and a huge clinical challenge without available effective treatment. P2 receptors family is involved in pain transmission and represents a promising target for pharmacological intervention. Traditional Chinese medicine (TCM) contains multiple components which are effective in targeting different pathological mechanisms involved in NP. Different from traditional analgesics, which target a single pathway, TCMs take the advantage of multiple components and multiple targets, and can significantly improve the efficacy of treatment and contribute to the prediction of the risks of NP. Compounds of TCM acting at nucleotide P2 receptors in neurons and glial cells could be considered as a potential research direction for moderating neuropathic pain. This review summarized the recently published data and highlighted several TCMs that relieved NP by acting at P2 receptors.

Enhanced fluorescence sensing of hydroxylated organotins by a boronic acid-linked Schiff base.

A simple Schiff base, 2-(2',4'-dihydroxybenzylidene)aminobenzeneboronic acid, was found to show a fluorescence enhancement in the presence of hydroxylated organotins in aqueous solution.

Pb(4)Br(11)(3-) cluster as a fluorescent indicator for micro water content in aprotic organic solvents.

Micro water content in aprotic solvents can be conveniently determined from the Stokes' shift value of the fluorescent anionic Pb(4)Br(11)(3-) cluster in situ formed in the test solvents.

Abnormal sympathetic activity after myocardial ischemia involving P2X4 in dorsal root ganglia.

The satellite glial cells (SGCs) of the dorsal root ganglia (DRG) expressed P2X4 receptor. In this study, we investigated the abnormal sympathetic activity after myocardial ischemia (MI) involving P2X4 receptor in the cervical DRG SGC. The results showed that MI injury upregulated the P2X4 receptor mRNA and protein in DRG, and the upregulated P2X4 receptor was co-localized with glial fibrillary acidic protein (GFAP) in DRG SGCs. P2X4 short hairpin RNA (shRNA) treatment decreased the expression of P2X4 receptor, counteracted the upregulation of GFAP and IL-1ß and inhibited P38MAPK phosphorylation in DRG of MI rats. These results indicate that application of P2X4 shRNA may reduce P2X4-mediated nociceptive signal via inhibiting DRG afferents to alleviate the abnormal sympathetic activity induced by MI.

Genome-wide screening and identification of novel proteolytic cleavage targets of caspase-8 and -10 in vitro.

Apoptosis executed by the mammalian caspase family plays a fundamental role in cellular homeostasis. Deregulation of this process is associated with several human diseases. The multimerization of ligand-induced death receptors results in the recruitment of the death inducing signaling complex and autocatalytic activation of initiator caspases, including caspase-8 and -10. However, it is still unclear how initiator caspases trigger and control the early apoptotic signaling pathways, partly because the downstream proteolytic cleavage targets of the initiator caspases are not completely known. Although it is known that a number of proteins are cleaved by various members of the caspase family, the identification of specific cleavage substrates of the initiator caspases 8 and 10, has been hindered by a lack of systematic and broadly applicable strategies for substrate identification. In the present study we constructed a mouse cDNA library and used it to perform a systematic, genome-wide screen for novel in vitro substrates of caspase-8 and -10. From this, we successfully identified six putative caspase substrates, including five novel proteins (ABCF1, AKAP1, CPE, DOPEY1 and GOPC1) that may be targeted specifically by the initiator caspases 8 and 10 during the early stages of apoptosis. These findings may provide useful information for elucidating the apoptotic signaling pathways downstream of the death receptors.