Global, regional, and national burden of ischaemic heart disease and its attributable risk factors in youth from 1990 to 2019: a Global Burden of Disease study.

OBJECTIVES: The objective of this study was to analyse the global, regional, and national burdens of ischaemic heart disease (IHD) in adults aged 15-49 years and its attributable risk factors from 1990 to 2019. STUDY DESIGN: Epidemiological study. METHODS: Data were obtained from the Global Burden of Disease (GBD) Study 2019. The estimated annual percentage change was used to evaluate temporal trends in incidence, deaths, and disability-adjusted life years (DALYs) of youth IHD. We selected IHD-associated risk factors, including five environmental/occupational factors, 16 behavioural risks, and five metabolic factors. We computed the age-standardised rates and percentage of age-standardised DALY rates attributable to these factors of youth IHD. RESULTS: Globally, there were 2.26 million cases of incidence, 0.63 million deaths, and 30.58 million DALYs in 2019. The age-standardised incidence, death, and DALY rates decreased from 1990 to 2019, whereas the absolute number of incidences, deaths, and DALYs increased significantly. Globally, approximately 94.1% of age-standardised DALY rates from IHD in youths aged 15-49 years are attributable to risk factors listed in the GBD 2019 dataset. The leading global and regional risk factors for youth IHD in 2019 were high low-density lipoprotein cholesterol (68.9%), high systolic blood pressure (51.2%), high body mass index (33.1%), smoking (30.5%), and ambient particulate-matter pollution (25.4%). CONCLUSIONS: The burden of IHD among young people is still heavy, and metabolic risk factors are the leading drivers of IHD. Therefore, formulating relevant policies to control and treat cardiovascular risk factors is an effective measure to reduce the IHD burden in youth.

Microbiota derived D-malate inhibits skeletal muscle growth and angiogenesis during aging via acetylation of Cyclin A.

Metabolites derived from the intestinal microbiota play an important role in maintaining skeletal muscle growth, function, and metabolism. Here, we found that D-malate (DMA) is produced by mouse intestinal microorganisms and its levels increase during aging. Moreover, we observed that dietary supplementation of 2% DMA inhibits metabolism in mice, resulting in reduced muscle mass, strength, and the number of blood vessels, as well as the skeletal muscle fiber type I/IIb ratio. In vitro assays demonstrate that DMA decreases the proliferation of vascular endothelial cells and suppresses the formation of blood vessels. In vivo, we further demonstrated that boosting angiogenesis by muscular VEGFB injection rescues the inhibitory effects of D-malate on muscle mass and fiber area. By transcriptomics analysis, we identified that the mechanism underlying the effects of DMA depends on the elevated intracellular acetyl-CoA content and increased Cyclin A acetylation rather than redox balance. This study reveals a novel mechanism by which gut microbes impair muscle angiogenesis and may provide a therapeutic target for skeletal muscle dysfunction in cancer or aging.

Specialized Retinal Endothelial Cells Modulate Blood-Retina Barrier in Diabetic Retinopathy.

Endothelial cells (EC) play essential roles in retinal vascular homeostasis. This study aimed to characterize retinal EC heterogeneity and functional diversity using single-cell RNA sequencing. Systematic analysis of cellular compositions and cell-cell interaction networks identified a unique EC cluster with high inflammatory gene expression in diabetic retina; sphingolipid metabolism is a prominent aspect correlated with changes in retinal function. Among sphingolipid-related genes, alkaline ceramidase 2 (ACER2) showed the most significant increase. Plasma samples of patients with nonproliferative diabetic retinopathy (NPDR) with diabetic macular edema (DME) or without DME (NDME) and active proliferative DR (PDR) were collected for mass spectrometry analysis. Metabolomic profiling revealed that the ceramide levels were significantly elevated in NPDR-NDME/DME and further increased in active PDR compared with control patients. In vitro analyses showed that ACER2 overexpression retarded endothelial barrier breakdown induced by ceramide, while silencing of ACER2 further disrupted the injury. Moreover, intravitreal injection of the recombinant ACER2 adeno-associated virus rescued diabetes-induced vessel leakiness, inflammatory response, and neurovascular disease in diabetic mouse models. Together, this study revealed a new diabetes-specific retinal EC population and a negative feedback regulation pathway that reduces ceramide content and endothelial dysfunction by upregulating ACER2 expression. These findings provide insights into cell-type targeted interventions for diabetic retinopathy.

Self-helped detection of obstructive sleep apnea based on automated facial recognition and machine learning.

PURPOSE: The diagnosis of obstructive sleep apnea (OSA) relies on time-consuming and complicated procedures which are not always readily available and may delay diagnosis. With the widespread use of artificial intelligence, we presumed that the combination of simple clinical information and imaging recognition based on facial photos may be a useful tool to screen for OSA. METHODS: We recruited consecutive subjects suspected of OSA who had received sleep examination and photographing. Sixty-eight points from 2-dimensional facial photos were labelled by automated identification. An optimized model with facial features and basic clinical information was established and tenfold cross-validation was performed. Area under the receiver operating characteristic curve (AUC) indicated the model's performance using sleep monitoring as the reference standard. RESULTS: A total of 653 subjects (77.2% males, 55.3% OSA) were analyzed. CATBOOST was the most suitable algorithm for OSA classification with a sensitivity, specificity, accuracy, and AUC of 0.75, 0.66, 0.71, and 0.76 respectively (P < 0.05), which was better than STOP-Bang questionnaire, NoSAS scores, and Epworth scale. Witnessed apnea by sleep partner was the most powerful variable, followed by body mass index, neck circumference, facial parameters, and hypertension. The model's performance became more robust with a sensitivity of 0.94, for patients with frequent supine sleep apnea. CONCLUSION: The findings suggest that craniofacial features extracted from 2-dimensional frontal photos, especially in the mandibular segment, have the potential to become predictors of OSA in the Chinese population. Machine learning-derived automatic recognition may facilitate the self-help screening for OSA in a quick, radiation-free, and repeatable manner.

IL-1ß knockdown inhibits cigarette smoke extract-induced inflammation and apoptosis in vascular smooth muscle cells.

OBJECTIVE: This study was aimed to investigate the role of interleukin-1ß (IL-1ß) in cigarette smoke extract (CSE)-induced apoptosis in vascular smooth muscle cells and the underlying mechanism in a rat derived cell line. METHODS: Rat thoracic aortic smooth muscle cells (A7r5) were divided into six groups including control, CSE (model), CSE+ overexpression empty vector (OvExp-EV), CSE+IL-1ß knockdown (KD), and CSE+ IL-1ß knockdown empty vector (KD-EV). The mRNA expression levels of IL-1ß and pregnancy-associated plasma protein A (PAPP-A) were detected by quantitative polymerase chain reaction (qPCR). The apoptosis of A7r5 cells was detected by flow cytometry. The expression levels of inflammatory mediators (TNFα, IL-6 and IL-8) and apoptotic proteins (Bax and Bcl-2) were determined by western blot. RESULTS: CSE induced significant apoptosis in vascular smooth muscle cells (P < 0.01) and elevated the mRNA levels of IL-1ß and PAPP-A (P < 0.01). CSE administration increased protein expression of Bax, TNF-α, IL-6, and IL-8, with significantly reduced Bcl-2 expression (P < 0.01). IL-1ß knockdown significantly decreased cell apoptosis via regulating the expression of these proteins (P < 0.05 or P < 0.01). CONCLUSION: IL-1ß is involved in CSE-induced PAPP-A expression and apoptosis in vascular smooth muscle cells, which might be considered as a target for preventing of cardiovascular diseases caused by cigarette smoking.

[Comparative study of microscope assisted minimally invasive anterior fusion and mobile microendoscopic discectomy assisted fusion for lumbar degenerative diseases].

Objective: To investigate the effectiveness of microscope assisted anterior lumbar discectomy and fusion (ALDF) and mobile microendoscopic discectomy assisted lumbar interbody fusion (MMED-LIF) for lumbar degenerative diseases. Methods: A clinical data of 163 patients with lumbar degenerative diseases who met the criteria between January 2018 and December 2020 was retrospectively analyzed. Fifty-three cases were treated with microscope assisted ALDF (ALDF group) and 110 cases with MMED-LIF (MMED-LIF group). There was no significant difference between the two groups in terms of gender, age, disease type, surgical segments, preoperative visual analogue scale (VAS) scores of low back pain and leg pain, Oswestry disability index (ODI), intervertebral space height, lordosis angle, and spondylolisthesis rate of the patients with lumbar spondylolisthesis ( P>0.05). The operation time, intraoperative blood loss, and hospital stay of the two groups were recorded. The effectiveness was evaluated by VAS scores of low back pain and leg pain and ODI. Postoperative lumbar X-ray films were taken to observe the position of Cage and measure the intervertebral space height, lordosis angle, and spondylolisthesis rate of the patients with lumbar spondylolisthesis. Results: The operations were successfully completed in both groups. The operation time, intraoperative blood loss, and hospital stay in ALDF group were less than those in MMED-LIF group ( P<0.05). The patients in both groups were followed up 12-36 months, with an average of 24 months. The VAS scores of low back pain and leg pain and ODI after operation were lower than those before operation in the two groups, and showed a continuous downward trend, with significant differences between different time points ( P<0.05). There were significant differences between two groups in VAS score of low back pain and ODI ( P<0.05) and no significant difference in VAS score of leg pain ( P>0.05) at each time point. The improvement rates of VAS score of low back pain and ODI in ALDF group were significantly higher than those in MMED-LIF group ( t=7.187, P=0.000; t=2.716, P=0.007), but there was no significant difference in the improvement rate of VAS score of leg pain ( t=0.556, P=0.579). The postoperative lumbar X-ray films showed the significant recovery of the intervertebral space height, lordosis angle, and spondylolisthesis rate at 2 days after operation when compared with preoperation ( P<0.05), and the improvements were maintained until last follow-up ( P>0.05). The improvement rates of intervertebral space height and lordosis angle in ALDF group were significantly higher than those in MMED-LIF group ( P<0.05). There was no significant difference in the reduction rate of spondylolisthesis between the two groups ( t=1.396, P=0.167). During follow-up, there was no loosening or breakage of the implant and no displacement or sinking of the Cage. Conclusion: Under appropriate indications, microscope assisted ALDF and MMED-LIF both can achieve good results for lumbar degenerative diseases. Microscope assisted ALDF was superior to MMED-LIF in the improvement of low back pain and function and the recovery of intervertebral space height and lordosis angle.

Disruption of USP9X in macrophages promotes foam cell formation and atherosclerosis.

Subendothelial macrophage internalization of modified lipids and foam cell formation are hallmarks of atherosclerosis. Deubiquitinating enzymes (DUBs) are involved in various cellular activities; however, their role in foam cell formation is not fully understood. Here, using a loss-of-function lipid accumulation screening, we identified ubiquitin-specific peptidase 9 X-linked (USP9X) as a factor that suppressed lipid uptake in macrophages. We found that USP9X expression in lesional macrophages was reduced during atherosclerosis development in both humans and rodents. Atherosclerotic lesions from macrophage USP9X-deficient mice showed increased macrophage infiltration, lipid deposition, and necrotic core content than control apolipoprotein E-KO (Apoe-/-) mice. Additionally, loss-of-function USP9X exacerbated lipid uptake, foam cell formation, and inflammatory responses in macrophages. Mechanistically, the class A1 scavenger receptor (SR-A1) was identified as a USP9X substrate that removed the K63 polyubiquitin chain at the K27 site. Genetic or pharmacological inhibition of USP9X increased SR-A1 cell surface internalization after binding of oxidized LDL (ox-LDL). The K27R mutation of SR-A1 dramatically attenuated basal and USP9X knockdown-induced ox-LDL uptake. Moreover, blocking binding of USP9X to SR-A1 with a cell-penetrating peptide exacerbated foam cell formation and atherosclerosis. In this study, we identified macrophage USP9X as a beneficial regulator of atherosclerosis and revealed the specific mechanisms for the development of potential therapeutic strategies for atherosclerosis.

RNA N6-methyladenosine modulates endothelial atherogenic responses to disturbed flow in mice.

Atherosclerosis preferentially occurs in atheroprone vasculature where human umbilical vein endothelial cells are exposed to disturbed flow. Disturbed flow is associated with vascular inflammation and focal distribution. Recent studies have revealed the involvement of epigenetic regulation in atherosclerosis progression. N6-methyladenosine (m6A) is the most prevalent internal modification of eukaryotic mRNA, but its function in endothelial atherogenic progression remains unclear. Here, we show that m6A mediates the epidermal growth factor receptor (EGFR) signaling pathway during EC activation to regulate the atherosclerotic process. Oscillatory stress (OS) reduced the expression of methyltransferase like 3 (METTL3), the primary m6A methyltransferase. Through m6A sequencing and functional studies, we determined that m6A mediates the mRNA decay of the vascular pathophysiology gene EGFR which leads to EC dysfunction. m6A modification of the EGFR 3' untranslated regions (3'UTR) accelerated its mRNA degradation. Double mutation of the EGFR 3'UTR abolished METTL3-induced luciferase activity. Adenovirus-mediated METTL3 overexpression significantly reduced EGFR activation and endothelial dysfunction in the presence of OS. Furthermore, thrombospondin-1 (TSP-1), an EGFR ligand, was specifically expressed in atheroprone regions without being affected by METTL3. Inhibition of the TSP-1/EGFR axis by using shRNA and AG1478 significantly ameliorated atherogenesis. Overall, our study revealed that METTL3 alleviates endothelial atherogenic progression through m6A-dependent stabilization of EGFR mRNA, highlighting the important role of RNA transcriptomics in atherosclerosis regulation.

Selective Retention of Bone Marrow Stromal Cells with Gelatin Sponge for Repair of Intervertebral Disc Defects after Microendoscopic Discectomy: A Prospective Controlled Study and 2-Year Follow-Up.

OBJECTIVE: Discectomy remains the classic procedure for treating lumbar intervertebral disc (IVD) herniation, but the occurrence of defects after discectomy is thought to be an important cause generating recurrent and accelerated IVD degeneration. Previous studies attempted suture of the annulus fissure, but the validity of this technique on restraining the degenerative process is controversial. On the other hand, cell therapies have been shown in multiple clinical and basic studies. Our purpose was to investigate the effectiveness of selective retention of autologous Bone Marrow Stromal Cells (BMSCs) with gelatin sponge in combination with annulus fibrosus suture (AFS) for the repair of IVD defects following mobile microendoscopic discectomy (MMED). METHODS: This prospective, two-armed, and controlled clinical study was conducted from December 2016 to December 2018. Written informed consent was obtained from each patient. Forty-five patients with typical symptoms, positive signs of radiculopathy, and obvious lumbar disc herniation observed by MRI were enrolled. Patients were divided into 3 groups with different treating methods: MMED (n = 15), MMED+AFS (n = 15), and MMED+AFS+BMSCs (n = 15). A postoperative 2-year follow-up was performed to evaluate the patient-reported outcomes of VAS, ODI, and SF-36. The improvement rate of VAS and ODI was calculated as [(latest-preoperative)/preoperative] to evaluate the therapeutic effect of the three groups. Assessment parameters included Pfirrmann grade, intervertebral disc height (IDH), and disc protrusion size (DPS), as measured by MRI to evaluate the morphological changes. RESULTS: All patients enrolled had a postoperative follow-up at 3, 6, 12, and 24 months. VAS and ODI scores were significantly improved compared to the preoperative status in all three groups with a mean DPS reduction rate over 50%. At the final follow-up, the improvement rate of the VAS score in the MMED+AFS+BMSCs group was significantly higher than the MMED+AFS and MMED groups (80.1% ± 7.6% vs. 71.3% ± 7.0% vs. 70.1% ± 7.8%), while ODI improvement showed a significant change (65.6% ± 8.8% vs. 59.9% ± 5.5% vs. 57.8% ± 8.1%). All participants showed significant improvement in SF-36 PCS and MCS; the differences between each group were not significant. The mean IDH loss rate of the MMED+AFS+BMSCs group was also significantly lower than other groups (-17.2% ± 1.3% vs. -27.6% ± 0.7% vs. -29.3% ± 2.2%). The Pfirrmann grade was aggravated in the MMED and MMED+AFS groups while maintained at the preoperative grade in the MMED+AFS+BMSCs group. No adverse events of cell transplantation or recurrence were found in all patients during the postoperative follow-up period. CONCLUSIONS: It is feasible and effective to repair lumbar IVD defects using SCR-enriched BMSCs with gelatin sponges, which warrants further study and development as a cell-based therapy for IVD repair.

hERG-deficient human embryonic stem cell-derived cardiomyocytes for modelling QT prolongation.

BACKGROUND: Long-QT syndrome type 2 (LQT2) is a common malignant hereditary arrhythmia. Due to the lack of suitable animal and human models, the pathogenesis of LQT2 caused by human ether-a-go-go-related gene (hERG) deficiency is still unclear. In this study, we generated an hERG-deficient human cardiomyocyte (CM) model that simulates 'human homozygous hERG mutations' to explore the underlying impact of hERG dysfunction and the genotype-phenotype relationship of hERG deficiency. METHODS: The KCNH2 was knocked out in the human embryonic stem cell (hESC) H9 line using the CRISPR/Cas9 system. Using a chemically defined differentiation protocol, we obtained and verified hERG-deficient CMs. Subsequently, high-throughput microelectrode array (MEA) assays and drug interventions were performed to characterise the electrophysiological signatures of hERG-deficient cell lines. RESULTS: Our results showed that KCNH2 knockout did not affect the pluripotency or differentiation efficiency of H9 cells. Using high-throughput MEA assays, we found that the electric field potential duration and action potential duration of hERG-deficient CMs were significantly longer than those of normal CMs. The hERG-deficient lines also exhibited irregular rhythm and some early afterdepolarisations. Moreover, we used the hERG-deficient human CM model to evaluate the potency of agents (nifedipine and magnesium chloride) that may ameliorate the phenotype. CONCLUSIONS: We established an hERG-deficient human CM model that exhibited QT prolongation, irregular rhythm and sensitivity to other ion channel blockers. This model serves as an important tool that can aid in understanding the fundamental impact of hERG dysfunction, elucidate the genotype-phenotype relationship of hERG deficiency and facilitate drug development.

Macrophage K63-Linked Ubiquitination of YAP Promotes Its Nuclear Localization and Exacerbates Atherosclerosis.

The Hippo/Yes-associated protein (YAP) pathway has pivotal roles in innate immune responses against pathogens in macrophages. However, the role of YAP in macrophages during atherosclerosis and its mechanism of YAP activation remain unknown. Here, we find that YAP overexpression in myeloid cells aggravates atherosclerotic lesion size and infiltration of macrophages, whereas YAP deficiency reduces atherosclerotic plaque. Tumor necrosis factor receptor-associated factor 6 (TRAF6), a downstream effector of interleukin-1ß (IL-1ß), triggers YAP ubiquitination at K252, which interrupts the interaction between YAP and angiomotin and results in enhanced YAP nuclear translocation. The recombinant IL-1 receptor antagonist anakinra reduces atherosclerotic lesion formation, which is abrogated by YAP overexpression. YAP level is increased in human and mouse atherosclerotic vessels, and plasma IL-1ß level in patients with STEMI is correlated with YAP protein level in peripheral blood mononuclear cells. These findings elucidate a mechanism of YAP activation, which might be a therapeutic target for atherosclerosis.

Retinol-binding protein 2 (RBP2) binds monoacylglycerols and modulates gut endocrine signaling and body weight.

Expressed in the small intestine, retinol-binding protein 2 (RBP2) facilitates dietary retinoid absorption. Rbp2-deficient (Rbp2-/- ) mice fed a chow diet exhibit by 6-7 months-of-age higher body weights, impaired glucose metabolism, and greater hepatic triglyceride levels compared to controls. These phenotypes are also observed when young Rbp2-/- mice are fed a high fat diet. Retinoids do not account for the phenotypes. Rather, RBP2 is a previously unidentified monoacylglycerol (MAG)-binding protein, interacting with the endocannabinoid 2-arachidonoylglycerol (2-AG) and other MAGs with affinities comparable to retinol. X-ray crystallographic studies show that MAGs bind in the retinol binding pocket. When challenged with an oil gavage, Rbp2-/- mice show elevated mucosal levels of 2-MAGs. This is accompanied by significantly elevated blood levels of the gut hormone GIP (glucose-dependent insulinotropic polypeptide). Thus, RBP2, in addition to facilitating dietary retinoid absorption, modulates MAG metabolism and likely signaling, playing a heretofore unknown role in systemic energy balance.

Interbody Fusion and Percutaneous Reduction For Lumbar Spondylolisthesis With Mobile Microendoscopic Discectomy Technique.

STUDY DESIGN: A minimally invasive surgical technique for lumbar spondylolisthesis. OBJECTIVE: The objective of this study was to investigate the feasibility and clinical efficacy of interbody fusion and percutaneous reduction for lumbar spondylolisthesis using mobile microendoscopic discectomy (MMED) technique. SUMMARY OF BACKGROUND DATA: Current surgical techniques for lumbar spondylolisthesis cause considerable trauma, so a minimally invasive technique is needed. MATERIALS AND METHODS: A total of 62 patients of lumbar spondylolisthesis (40 patients of degenerative spondylolisthesis and 22 patients of isthmic spondylolisthesis) were treated with interbody fusion and percutaneous reduction using the MMED technique. A 2.5-cm longitudinal incision was made on the side of dominant symptoms, with paraspinous approach used for degenerative spondylolisthesis and transforaminal approach for isthmic spondylolisthesis. The fenestration and decompression were performed under MMED. The intervertebral space was released through an outer tube under direct vision, followed by bone graft and cage implantation. Percutaneous pedicle screws were used, with the residual spondylolisthesis further reduced. The patients were followed up to evaluate the clinical results. RESULTS: The procedure was successful in all patients. Postoperative radiographs showed sufficient decompression and improvement of spinal alignment for both groups. The average reduction rate of the spondylolisthesis was 68% for degenerative spondylolisthesis group and 66% for isthmic spondylolisthesis group. The patients of 2 groups were followed up for 12-24 months. At the final follow-up, the Oswestry Disability Index and Visual Analogue Scale scores decreased significantly compared with preoperation for both groups. The clinical results were excellent in 22 cases, good in 16 and fair in 2 for degenerative spondylolisthesis group, and excellent in 11 cases, good in 10 and fair in 1 for isthmic spondylolisthesis group according to the Macnab Scale. CONCLUSION: Interbody fusion and percutaneous reduction with MMED provides a minimally invasive procedure for lumbar spondylolisthesis, with sufficient decompression, reduction, fusion, and satisfactory clinical results.

Interrogation of nonconserved human adipose lincRNAs identifies a regulatory role of linc-ADAL in adipocyte metabolism.

Long intergenic noncoding RNAs (lincRNAs) have emerged as important modulators of cellular functions. Most lincRNAs are not conserved among mammals, raising the fundamental question of whether nonconserved adipose-expressed lincRNAs are functional. To address this, we performed deep RNA sequencing of gluteal subcutaneous adipose tissue from 25 healthy humans. We identified 1001 putative lincRNAs expressed in all samples through de novo reconstruction of noncoding transcriptomes and integration with existing lincRNA annotations. One hundred twenty lincRNAs had adipose-enriched expression, and 54 of these exhibited peroxisome proliferator-activated receptor γ (PPARγ) or CCAAT/enhancer binding protein α (C/EBPα) binding at their loci. Most of these adipose-enriched lincRNAs (~85%) were not conserved in mice, yet on average, they showed degrees of expression and binding of PPARγ and C/EBPα similar to those displayed by conserved lincRNAs. Most adipose lincRNAs differentially expressed (n = 53) in patients after bariatric surgery were nonconserved. The most abundant adipose-enriched lincRNA in our subcutaneous adipose data set, linc-ADAL, was nonconserved, up-regulated in adipose depots of obese individuals, and markedly induced during in vitro human adipocyte differentiation. We demonstrated that linc-ADAL interacts with heterogeneous nuclear ribonucleoprotein U (hnRNPU) and insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2) at distinct subcellular locations to regulate adipocyte differentiation and lipogenesis.

Bilateral decompression and intervertebral fusion via unilateral fenestration for complex lumbar spinal stenosis with a mobile microendoscopic technique.

For complex lumbar spinal stenosis, using of endoscopy technique may provide clear vision with less invasive dissection of paravertebral muscle. The objective of this study was to evaluate the feasibility and clinical efficacy of bilateral decompression and intervertebral fusion via unilateral fenestration for complex lumbar spinal stenosis using mobile microendoscopic discectomy (MMED) technique.A total of 61 patients with complex lumbar spinal stenosis (lumbar canal stenosis combined with degenerative spondylolisthesis, instability, and scoliosis) were treated with this procedure. Patients with isolated lumbar spinal stenosis or spondylolisthesis greater than grade II were excluded. The index levels included L4/5 in 52 patients, L5/S1 in 6 patients, L3-L5 in 2 patients and L4-S1 in 1 patient. The preoperative Oswestry Disability Index (ODI) score was 42.6 ±â€Š10.2, lumbar visual analog scale (VAS) score was 6.1 ±â€Š4.2, and leg VAS score was 7.1 ±â€Š5.1. During the operation, ipsilateral enlarged fenestration was made using the MMED technique. The disc and cartilage endplate were thoroughly removed, and the contralateral ligamentum flavum and the inner layer of lamina were undercut to release the contralateral nerve root. The intervertebral space was released and prepared, followed by bone grafting and cage insertion. Percutaneous pedicle system was used for reduction and fixation. The operative time and blood loss were recorded, and patients were followed-up for at least 3 years (36-48 months, average 41 months) to evaluate the clinical efficacy.The procedure was successful in all patients, with no nerve injury or conversion to open operation. The mean operative time was 120 minutes (range, 100-180 minutes), with a mean blood loss of 100 mL (range, 50-200 mL). Postoperative x-ray and CT showed sufficient decompression and improvement of spinal alignment. At 3 years after surgery, the ODI scores, lumbar and leg VAS scores decreased from preoperative 42.6 ±â€Š10.2, 6.1 ±â€Š4.2, and 7.1 ±â€Š5.1 to 8.6 ±â€Š7.0, 1.8 ±â€Š1.3, and 0.9 ±â€Š0.6, respectively (P = .00 for each comparison). The clinical results were excellent in 36 cases, good in 23, and fair in 2, according to the MacNab scale.The procedure of bilateral decompression and intervertebral fusion via unilateral fenestration using the MMED technique can provide satisfactory clinical results for complex lumbar spinal stenosis.

CRISPR/Cas9-Mediated Gene Editing in Human iPSC-Derived Macrophage Reveals Lysosomal Acid Lipase Function in Human Macrophages-Brief Report.

OBJECTIVE: To gain mechanistic insights into the role of LIPA (lipase A), the gene encoding LAL (lysosomal acid lipase) protein, in human macrophages. APPROACH AND RESULTS: We used CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9) technology to knock out LIPA in human induced pluripotent stem cells and then differentiate to macrophage (human-induced pluripotent stem cells-derived macrophage [IPSDM]) to explore the human macrophage LIPA loss-of-function phenotypes. LIPA was abundantly expressed in monocyte-derived macrophages and was markedly induced on IPSDM differentiation to comparable levels as in human monocyte-derived macrophage. IPSDM with knockout of LIPA (LIPA-/-) had barely detectable LAL enzymatic activity. Control and LIPA-/- IPSDM were loaded with [3H]-cholesteryl oleate-labeled AcLDL (acetylated low-density lipoprotein) followed by efflux to apolipoprotein A-I. Efflux of liberated [3H]-cholesterol to apolipoprotein A-I was abolished in LIPA-/- IPSDM, indicating deficiency in LAL-mediated lysosomal cholesteryl ester hydrolysis. In cells loaded with [3H]-cholesterol-labeled AcLDL, [3H]-cholesterol efflux was, however, not different between control and LIPA-/- IPSDM. ABCA1 (ATP-binding cassette, subfamily A, member 1) expression was upregulated by AcLDL loading but to a similar extent between control and LIPA-/- IPSDM. In nonlipid loaded state, LIPA-/- IPSDM had high levels of cholesteryl ester mass compared with minute amounts in control IPSDM. Yet, with AcLDL loading, overall cholesteryl ester mass was increased to similar levels in both control and LIPA-/- IPSDM. LIPA-/- did not impact lysosomal apolipoprotein-B degradation or expression of IL1B, IL6, and CCL5. CONCLUSIONS: LIPA-/- IPSDM reveals macrophage-specific hallmarks of LIPA deficiency. CRISPR/Cas9 and IPSDM provide important tools to study human macrophage biology and more broadly for future studies of disease-associated LIPA genetic variation in human macrophages.

Hydroxyeicosapentaenoic acids and epoxyeicosatetraenoic acids attenuate early occurrence of nonalcoholic fatty liver disease.

BACKGROUND AND PURPOSE: The ω-3 polyunsaturated fatty acids (PUFAs) mediate protective effects on several metabolic disorders. However, the functions of their metabolites in the early stage of nonalcoholic fatty liver disease (NAFLD) are largely unknown. EXPERIMENTAL APPROACH: Mice were fed a control diet, high-fat diet (HFD) or ω-3 PUFA-enriched HFD (ω3HFD) for 4 days and phenotypes were analysed. LC-MS/MS was used to determine the eicosanoid profiles. Primary hepatocytes and peritoneal macrophages were used for the mechanism study. KEY RESULTS: In short-term HFD-fed mice, the significantly increased lipid accumulation in the liver was reversed by ω-3 PUFA supplementation. Metabolomics showed that the plasma concentrations of hydroxyeicosapentaenoic acids (HEPEs) and epoxyeicosatetraenoic acids (EEQs) were reduced by a short-term HFD and markedly increased by the ω3HFD. However, HEPE/EEQ treatment had no direct protective effect on hepatocytes. ω3HFD also significantly attenuated HFD-induced adipose tissue inflammation. Furthermore, the expression of pro-inflammatory cytokines and activation of the JNK pathway induced by palmitate were suppressed by HEPEs and EEQs in macrophages. 17,18-EEQ, 5-HEPE and 9-HEPE were identified as the effective components among these metabolites, as indicated by their greater suppression of the palmitate-induced expression of inflammatory factors, chemotaxis and JNK activation compared to other metabolites in macrophages. A mixture of 17,18-EEQ, 5-HEPE and 9-HEPE significantly ameliorated the short-term HFD-induced accumulation of macrophages in adipose tissue and hepatic steatosis. CONCLUSION AND IMPLICATIONS: 17,18-EEQ, 5-HEPE and 9-HEPE may be potential approaches to prevent NAFLD in the early stage by inhibiting the inflammatory response in adipose tissue macrophages via JNK signalling.

A model of acute central cervical spinal cord injury syndrome combined with chronic injury in goats.

PURPOSE: To develop a large animal model for acute central cervical spinal cord injury syndrome (ACCSCIS. METHODS: Twenty-four adult male goats were randomized into four groups including group A with acute compression injury, group B with anterior chronic compression, group C as the test group that received anterior chronic compression by screw and acute compression by posterior balloon insertion, and group D as normal controls that received sham surgery. Neurological function (modified Tarlov motor function), CT, MRI, cortical somatosensory evoked potentials (CSEP), and pathological analysis were evaluated. The data were analyzed statistically. RESULTS: The motor function of the goats in group C was significantly lower than other groups. CSEP before spinal cord compression showed a stable pattern. Spinal cord compression resulted in a gradual decrement in the peak latency and significant increment in the peak amplitude. Cervical spinal canal occupying ratio was significantly lower in group C than the other groups. MRI revealed focal low signal in T1 weighted images and focal high signal in T2 weighted images in group C. Pathological analysis showed more severe lesions in the gray matter than that in the white matter in group C. CONCLUSIONS: The model well simulated the pathogenesis and resembled the clinical characteristics of ACCSCIS. This model seems to have the potential to contribute to the development of effective therapies for ACCSCIS.

Thoracic Endoscopic-Assisted Mini-Open Surgery for Thoracic and Thoracolumbar Spinal Cord Compression.

Intervertebral disc herniation is a common cause of spinal cord compression, especially for the thoracic and thoracolumbar spinal cord, which has limited buffer space in the spinal canal. Spinal cord compression usually causes decreased sensation and paralysis of limbs below the level of compression, urinary and fecal incontinence, and/or urinary retention, which brings great suffering to the patients and usually requires surgical intervention. Thoracotomy or abdominothoracic surgery is usually performed for the thoracolumbar cord compression caused by hard intervertebral disc herniation. However, there is high risk of trauma and complications with this surgery. To reduce the surgical trauma and obtain good visibility, we designed athoracic endoscopic-assisted mini-open surgery for thoracic and thoracolumbar disc herniation, and performed this procedure on 10 patients who suffered from hard thoracic or thoracolumbar spinal cord compression. During the procedure, the thoracic endoscopy provided clear vision of the surgical field with a good light source. The compression could be fully exposed and completely removed, and no nerve root injury or spinal cord damage occurred. All patients achieved obvious recovery of neurological function after this procedure. This technique possesses the merits of minimal trauma, increased safety, and good clinical results. The aim of this study is to introduce this thoracic endoscopic-assisted mini-open surgery technique, and we believe that this technique will be a good choice for the thoracic and thoracolumbar cord compression caused by hard intervertebral disc herniation.

Adipocyte-specific overexpression of retinol-binding protein 4 causes hepatic steatosis in mice.

There is considerable evidence that both retinoids and retinol-binding protein 4 (RBP4) contribute to the development of liver disease. To understand the basis for this, we generated and studied transgenic mice that express human RBP4 (hRBP4) specifically in adipocytes. When fed a chow diet, these mice show an elevation in adipose total RBP4 (mouse RBP4 + hRBP4) protein levels. However, no significant differences in plasma RBP4 or retinol levels or in hepatic or adipose retinoid (retinol, retinyl ester, and all-trans-retinoic acid) levels were observed. Strikingly, male adipocyte-specific hRBP4 mice fed a standard chow diet display significantly elevated hepatic triglyceride levels at 3-4 months of age compared to matched littermate controls. When mice were fed a high-fat diet, this hepatic phenotype, as well as other metabolic phenotypes (obesity and glucose intolerance), worsened. Because adipocyte-specific hRBP4 mice have increased tumor necrosis factor-α and leptin expression and crown-like structures in adipose tissue, our data are consistent with the notion that adipose tissue is experiencing RBP4-induced inflammation that stimulates increased lipolysis within adipocytes. Our data further establish that elevated hepatic triglyceride levels result from increased hepatic uptake of adipose-derived circulating free fatty acids. We obtained no evidence that elevated hepatic triglyceride levels arise from increased hepatic de novo lipogenesis, decreased hepatic free fatty acid oxidation, or decreased very-low-density lipoprotein secretion. CONCLUSION: Our investigations establish that RBP4 expressed in adipocytes induces hepatic steatosis arising from primary effects occurring in adipose tissue. (Hepatology 2016;64:1534-1546).