Comparative transcriptome analysis reveals the function of SlPRE2 in multiple phytohormones biosynthesis, signal transduction and stomatal development in tomato.

KEY MESSAGE: Transcriptomic, physiological, and qRT-PCR analysis revealed the potential mechanism by which SlPRE2 regulates plant growth and stomatal size via multiple phytohormone pathways in tomato. Paclobutrazol resistance proteins (PREs) are atypical members of the basic/helix-loop-helix (bHLH) transcription factor family that regulate plant morphology, cell size, pigment metabolism and abiotic stress in response to different phytohormones. However, little is known about the network regulatory mechanisms of PREs in plant growth and development in tomato. In this study, the function and mechanism of SlPRE2 in tomato plant growth and development were investigated. The quantitative RT-PCR results showed that the expression of SlPRE2 was regulated by multiple phytohormones and abiotic stresses. It showed light-repressed expression during the photoperiod. The RNA-seq results revealed that SlPRE2 regulated many genes involved in photosynthesis, chlorophyll metabolism, phytohormone metabolism and signaling, and carbohydrate metabolism, suggesting the role of SlPRE2 in gibberellin, brassinosteroid, auxin, cytokinin, abscisic acid and salicylic acid regulated plant development processes. Moreover, SlPRE2 overexpression plants showed widely opened stomata in young leaves, and four genes involved in stomatal development showed altered expression. Overall, the results demonstrated the mechanism by which SlPRE2 regulates phytohormone and stress responses and revealed the function of SlPRE2 in stomatal development in tomato. These findings provide useful clues for understanding the molecular mechanisms of SlPRE2-regulated plant growth and development in tomato.

Proper mechanical stress promotes femoral head recovery from steroid-induced osteonecrosis in rats through the OPG/RANK/RANKL system.

BACKGROUND: Long-term use of steroid may lead to osteonecrosis of the femoral head (ONFH). Mechanical stress may help bone formation and remodeling. This study aimed to probe the role of mechanical stress in the femoral head recovery in rats. METHODS: Rat models with ONFH were induced by steroid. Rats were subjected to different levels of mechanical stress (weight-bearing training), and then the morphology and bone density of femoral head of rats were measured. The mRNA and protein levels of the OPG/RANK/RANKL axis in rat femoral head were assessed. Gain- and loss-of function experiments of OPG were performed to identify its role in femoral head recovery following stress implement. The ex vivo cells were extracted and the effects of stress and OPG on osteogenesis in vitro were explored. RESULTS: Steroid-induced ONFH rats showed decreased bone density and increased bone spaces, as well as necrotic cell colonies and many cavities in the cortical bones and trabeculars. Proper mechanical stress or upregulation of OPG led to decreased RANK/RANKL expression and promoted femoral head recovery from steroid-induced osteonecrosis. However, excessive mechanical stress might impose too much load on the femurs thus leading even retard femoral head recovery process. In addition, the in vitro experimental results supported that proper stress and overexpression of OPG increased the osteogenesis of ex vivo cells of femoral head. CONCLUSION: This study provided evidence that proper mechanical stress promoted femoral head recovery from steroid-induced osteonecrosis through the OPG/RANK/RANKL system, while overload might inhibit the recovery process. This study may offer novel insights for ONFH treatment.

A case of polyglucosan body myopathy caused by an RBCK1 gene variant and literature review.

OBJECTIVE: To analyze the clinical and genetic characteristics of a patient with Polyglucosan body myopathy 1 (PGBM1) caused by a novel compound heterozygous variant in the RBCK1 gene. METHODS: The clinical data of the patient were collected, next-generation sequencing technology was used to determine the exome sequence of the patient, and the suspected pathogenic locus was verified by Sanger sequencing. RESULTS: Through whole-exome sequencing, we found that there were c.919G>T; p. (Glu307*) and c.723_730dup; p. (Glu244fs) variants of the RBCK1 gene in the patient, inherited from his parents, constituting a compound heterozygous variation. According to the guidelines of the American College of Medical Genetics and Genomics (ACMG), the two variants were rated as pathogenic, but there were no comparable cases. Previous literature reported 24 patients with RBCK1 gene variants, involving a total of 20 myocardial and 18 skeletal muscle cases. CONCLUSIONS: The patient was twice diagnosed with cardiac insufficiency, neglecting the usual manifestations of muscle weakness, resulting in misdiagnosis. Later, novel variants in the RBCK1 gene were discovered through whole-exome sequencing, and symptomatic treatment was given after diagnosis. The importance of whole-exome sequencing technology in disease diagnosis and genetic counseling was emphasized.

A Proactive Intervention Study in Metabolic Syndrome High-Risk Populations Using Phenome-Based Actionable P4 Medicine Strategy.

The integration of predictive, preventive, personalized, and participatory (P4) healthcare advocates proactive intervention, including dietary supplements and lifestyle interventions for chronic disease. Personal profiles include deep phenotypic data and genetic information, which are associated with chronic diseases, can guide proactive intervention. However, little is known about how to design an appropriate intervention mode to precisely intervene with personalized phenome-based data. Here, we report the results of a 3-month study on 350 individuals with metabolic syndrome high-risk that we named the Pioneer 350 Wellness project (P350). We examined: (1) longitudinal (two times) phenotypes covering blood lipids, blood glucose, homocysteine (HCY), and vitamin D3 (VD3), and (2) polymorphism of genes related to folic acid metabolism. Based on personalized data and questionnaires including demographics, diet and exercise habits information, coaches identified 'actionable possibilities', which combined exercise, diet, and dietary supplements. After a 3-month proactive intervention, two-thirds of the phenotypic markers were significantly improved in the P350 cohort. Specifically, we found that dietary supplements and lifestyle interventions have different effects on phenotypic improvement. For example, dietary supplements can result in a rapid recovery of abnormal HCY and VD3 levels, while lifestyle interventions are more suitable for those with high body mass index (BMI), but almost do not help the recovery of HCY. Furthermore, although people who implemented only one of the exercise or diet interventions also benefited, the effect was not as good as the combined exercise and diet interventions. In a subgroup of 226 people, we examined the association between the polymorphism of genes related to folic acid metabolism and the benefits of folate supplementation to restore a normal HCY level. We found people with folic acid metabolism deficiency genes are more likely to benefit from folate supplementation to restore a normal HCY level. Overall, these results suggest: (1) phenome-based data can guide the formulation of more precise and comprehensive interventions, and (2) genetic polymorphism impacts clinical responses to interventions. Notably, we provide a proactive intervention example that is operable in daily life, allowing people with different phenome-based data to design the appropriate intervention protocol including dietary supplements and lifestyle interventions. Supplementary Information: The online version contains supplementary material available at 10.1007/s43657-023-00115-z.

A community effort to assess and improve computerized interpretation of 12-lead resting electrocardiogram.

Computerized interpretation of electrocardiogram plays an important role in daily cardiovascular healthcare. However, inaccurate interpretations lead to misdiagnoses and delay proper treatments. In this work, we built a high-quality Chinese 12-lead resting electrocardiogram dataset with 15,357 records, and called for a community effort to improve the performances of CIE through the China ECG AI Contest 2019. This dataset covers most types of ECG interpretations, including the normal type, 8 common abnormal types, and the other type which includes both uncommon abnormal and noise signals. Based on the Contest, we systematically assessed and analyzed a set of top-performing methods, most of which are deep neural networks, with both their commonalities and characteristics. This study establishes the benchmarks for computerized interpretation of 12-lead resting electrocardiogram and provides insights for the development of new methods. Graphical Abstract A community effort to assess and improve computerized interpretation of 12-lead resting electrocardiogram.

LncRNA NORAD promotes bone marrow stem cell differentiation and proliferation by targeting miR-26a-5p in steroid-induced osteonecrosis of the femoral head.

BACKGROUND: Steroid-induced osteonecrosis of the femoral head (SONFH) is a devastating orthopedic disease, which seriously affects the quality of life of patients. The study aims to investigate the effects of LncRNA NORAD on SONFH. METHODS: Human bone marrow-derived mesenchymal stem cells (hBMSCs) were isolated from the proximal femur of patients during routine orthopedic surgery and then cultured with dexamethasone (Dex) and transfected with NORAD overexpression vector, siRNA-NORAD and miR-26a-5p mimics. The mRNA expression of NORAD, miR-26a-5p, OPG, RANK, and RANKL was detected by RT-qPCR. Cell proliferation and apoptosis was measured by CCK-8 assay and flow cytometry, respectively. The protein expression of RUNX2, OPG, RANK, and RANKL was detected by western blot. The dual-luciferase reporter gene assay was performed to confirm the binding between NORAD and miR-26a-5p. RESULTS: NORAD expression was downregulated in SONFH tissues, while miR-26a-5p expression was upregulated. Overexpression of NORAD improved DEX-induced inhibition of proliferation and differentiation, and promotion of apoptosis in hBMSCs, while knockdown of NORAD led to the opposite results. Moreover, NORAD improved DEX-induced inhibition of proliferation and differentiation, and promotion of apoptosis by regulation of miR-26a-5p in hBMSCs. CONCLUSIONS: NORAD expression was downregulated in SONFH tissues, while miR-26a-5p expression was upregulated. NORAD improved DEX-induced inhibition of proliferation and differentiation, and promotion of apoptosis by regulation of miR-26a-5p in hBMSCs.

Thrombolysis Combined Therapy Using CuS@SiO2-PEG/uPA Nanoparticles.

Massive hemorrhage caused by the uncontrolled release of thrombolysis drugs is a key issue of thrombolysis therapy in clinical practice. In this study, we report a near-infrared (NIR) light-triggered drug delivery system, i.e., CuS@mSiO2-PEG (CSP) nanoparticles, for the loading of a thrombolytic drug (urokinase plasminogen activators, uPA). CSP nanoparticles with the CuS nanoparticles as photothermal agents and mesoporous SiO2 for the loading of uPA were synthesized using a facile hydrothermal method. The CSP core-shell nanoparticles were demonstrated to possess excellent photothermal performance, exhibiting a photothermal conversion efficiency of up to 52.8%. Due to the mesoporous SiO2 coating, the CSP core-shell nanoparticles exhibited appropriate pore size, high pore volume, and large surface area; thus, they showed great potential to be used as drug carriers. Importantly, the release of uPA from CuS@mSiO2-PEG/uPA (CSPA) carriers can be promoted by the NIR laser irradiation. The drug loading content of uPA for the as-prepared NIR-triggered drug delivery system was calculated to be 8.2%, and the loading efficiency can be determined to be as high as 89.6%. Due to the excellent photothermal effect of CSP nanocarriers, the NIR-triggered drug delivery system can be used for infrared thermal imaging in vivo. The in vivo thrombolysis assessment demonstrated that the NIR-triggered drug delivery system showed excellent thrombolytic ability under the irradiation of an 808 nm laser, showing the combined therapy for thrombolysis. As far as we know, the CSPA core-shell nanoparticles used as NIR-triggered drug delivery systems for thrombolysis have not been reported.

Finite element study on whether posterior upper wall fracture is a risk factor for the failure of short-segment pedicle screw fixation in the treatment of L1 burst fracture.

PURPOSE: To establish the finite element model of T12 and L2 (T12-L2) pedicle screw fixation for severe L1 burst fracture, and quantitatively simulate and analyze the screw stress and vertebral displacement in different degrees of L1 posterior upper wall fracture (PUWF), and evaluate whether PUWF degree is a risk factor for fixation failure. METHODS: The data of 6 healthy volunteers were used to establish a finite element model of T12-L2 pedicle screw fixation for type A severe burst fractures. The stress and displacement of the conventional and Schanz pedicle screws for the different degrees of PUWF (including the anterior upper wall of the vertebral canal and the bipedicle) were evaluated. RESULTS: The maximum stress and L1 displacement of conventional and Schanz pedicle screws were positively correlated with the severity of the PUWF (P<0.05). During anterior flexion, the maximum stress of conventional pedicle screws for 70% type I were 538.3±59.75MPa and the maximum stress of Schanz pedicle screws for 90% type Ⅱ, 90% type Ⅲ and 70% type IV fractures were close to the fatigue threshold. The maximum stress during anterior flexion were significantly higher than those during posterior extension, bending and rotation (P<0.05). CONCLUSION: The posterior upper wall fracture of vertebral body (VB) of type A burst fracture is not an independent risk factor for the failure of short-segment pedicle screw fixation (SSPSF). Anterior flexion of type A fractures combined with severe PUWF of VB was a risk factor for the failure of SSPSF.

Biomechanical effects of USS fixation with different screw insertion depths on the vertebrae stiffness and screw stress for the treatment of the L1 fracture.

OBJECTIVE: To evaluate the biomechanical effects of internal fixation with different screw insertion depths on vertebrae stiffness and screw stress for L1 fracture. METHODS: The established L1 fracture was fixed with 10 different depths of screw insertion: 10-100% screw-path length (SPL). Loading on the T12 endplate was simulated. RESULTS: Screws inserted to 60-100% depths has a higher axial displacement of screw against injured vertebrae and maximum stress of screws compared to those of screws inserted to 30-50% depths and 10-20% (P< 0.05). No significant difference was noted among 60-100% SPL groups. Under single loading condition, the incidence rate of maximum stress of each screw ranged from 16.7-50.0%. Chi-square test showed superior screw has a higher incidence rate of maximum stress than inferior screw (P< 0.05). CONCLUSIONS: Screws inserted to 60% depth or more can achieve effective strength to withstand the postoperative height correction loss of the L1 vertebrae fracture. However, continuous prolonged depth of screw insertion did not significantly increase the effective strength of the screw against injured vertebrae and maximum equivalent stress of screws. The incidence rate of the maximum stress of each screw in correlated with position of screw insertion but not associated with the screw insertion depth.

Real-Time ECG Delineation with Randomly Selected Wavelet Transform Feature and Random Walk Estimation.

Detection of Electrocardiogram (ECG) characteristic points can provide critical diagnostic information about heart diseases. We propose a novel feature extraction and machine learning scheme for ECG delineation. A new feature, termed as randomly selected wavelet transform (RSWT), is proposed to effectively represent ECG morphology. With the RSWT feature pool, a regression tree is trained to estimate the probability distribution to the direction toward the target point, relative to the current position. The continual random walk through 1D space will eventually produce a reliable region from which the final position of the target point is derived. The evaluation results on QT database show better detection accuracy compared with other studies while providing real-time processing capability.

Modified Posterior Short-Segment Pedicle Screw Instrumentation for Lumbar Burst Fractures with Incomplete Neurological Deficit.

PURPOSE: We have introduced a method of modified posterior short-segment pedicle screw fixation and evaluated its clinical effects in treating lumbar burst fractures with incomplete neurological deficits. METHODS: The data from 22 patients with lumbar burst fracture and incomplete neurological deficits who had undergone modified posterior short-segment instrumentation with Schanz screw fixation from January 2012 to February 2018 in our clinic were evaluated in the present retrospective study. All Schanz screws were implanted in an oblique downward direction into the vertebrae above and below the injured vertebra (insertion depth, 90%-100%). The implants were removed ∼1 year after surgery. Neurological function, back pain, anterior and posterior body height ratio, kyphosis angle, percentage of canal compromise, fracture severity, and treatment-related complications were evaluated. RESULTS: Technical success was achieved in all 22 patients. No infection, instrument loosening or failure, or breakage was observed. Statistically significant improvements with regard to the anterior body height (P < 0.05) and posterior body height (P < 0.05) ratios, kyphosis angle, and percentage of canal compromise (P < 0.05) were observed at 1 week postoperatively or the final follow-up visit. No correction loss had occurred at the final follow-up examination. Postoperatively, all patients with neurological deficits had functional improvement equivalent to ≥1 grade on the American Spinal Injury Association impairment scale and fracture union. Back pain was greatly improved postoperatively. CONCLUSIONS: Short-segment Schanz screw fixation implanted in an oblique downward direction seems to be a promising method for lumbar burst fractures with incomplete neurological deficits because it provided good clinical and radiographic outcomes.

Oleic/Palmitate Induces Apoptosis in Human Articular Chondrocytes via Upregulation of NOX4 Expression and ROS Production.

The association between obesity and reactive oxygen species (ROS) production in osteoarthritis (OA) patients has already been identified. However, the specific mechanism underlying ROS production and OA progression has never been elucidated. Osteoarthritic cartilage was obtained from patients undergoing total hip arthroplasty, and chondrocytes were isolated from these tissues. The cells were treated with varying concentrations (10,100,500 µM, and 5 mM) of oleic/palmitate (O/P) mixture at different times, that is at 6, 24, and 48 h. Cell viability was determined using MTT assay. ROS production was detected using immunofluorescence and flow cytometry. The protein levels of NOX4 and cleaved-caspase3 were detected using Western blot assay. O/P significantly decreased cell viability at 10, 100, 500 µM, and 5 mM in a dose-dependent manner. Furthermore, the cell viability was reduced by 500 µM O/P mixture at 6, 24, and 48 h in a time-dependent manner. Pretreatment with 500 µM O/P significantly enhanced ROS production and cell apoptosis in chondrocytes. Furthermore, treatment with O/P mixture significantly enhanced the expression of NOX4 and caspase3 activation in a dose- and time- dependent manner. More importantly, inhibition of NOX4 could partially eliminate O/P-induced chondrocytes apoptosis by reducing ROS production. To conclude, O/P treatment enhances ROS production and cell apoptosis mainly by upregulating the protein levels of NOX4 and caspase3 activation in chondrocytes, indicating a potential therapeutic target of OA in obesity patients.

Association between the vitamin D receptor gene polymorphism and osteoporosis.

The influence of the vitamin D receptor (VDR) gene for the risk of osteoporosis remains to be elucidated. The aim of the present study was to understand the distribution of various single-nucleotide polymorphisms (SNPs) within the VDR gene and its association with the risk of osteoporosis. In total, 378 subjects without a genetic relationship were recruited to the study between January 2013 and July 2015. The subjects were divided into three groups, which were the normal (n=234), osteoporosis (n=65) and osteoporosis with osteoporotic fracture (n=79) groups. Three pertinent SNPs of the VDR gene rs17879735 (ApaI, Allele A/a, SNP C>A) were examined with polymerase chain reaction-restriction fragment length polymorphism. The bone mineral density (BMD) of the lumbar spine (L2-L4), femoral neck, Ward's and Tro was measured using dual-energy X-ray absorptiometry. The distributions of genotype frequencies aa, AA and Aa were 48.68, 42.86 and 8.46%, separately. Following analysis of each site, BMD, body mass index (BMI) and age, BMD for each site was negatively correlated with age (P<0.01) and positively correlated with BMI (P<0.01). Correction analysis revealed that there were significant differences in the Ward's triangle BMD among each genotype (P<0.05), in which the aa genotype exhibited the lower BMD (P<0.05). No significant difference was identified among the different genotypes in the occurrence of osteoporosis with osteoporotic fracture (P>0.05). In conclusion, these indicated that the VDR gene ApaI polymorphisms had an important role in the osteoporosis risk.

Prospective and comparative analysis of internal fixation of femoral neck fractures with or without vascularized iliac graft in young adults.

The purpose of this study was to compare the effectiveness of 2 surgical approaches for femoral neck fractures in young adults: internal fixation with or without a vascularized iliac graft. Between January 1998 and December 2008, seventy-eight patients presented with a Garden type III (n=38) or IV (n=40) femoral neck fracture. Thirty-eight patients were women and 40 were men, with an average age of 28 years (range, 16-38 years). Fractures were caused by fall injury (n=24), motor vehicle accident (n=36), and heavy weight lifting (n=18). Patients were randomly divided into 2 groups. Group A underwent internal fixation with 2 cannulated compression screws combined with an iliac graft supported by the ascending branch of the lateral femoral circumflex artery (n=44), and group B underwent internal fixation with 3 cannulated compression screws (n=34). Average follow-up was 4.5 years (range, 2-8 years), and mean Harris Hip Score was 92 (range, 62-100) in group A and 84 (range, 40-100) in group B. Average fracture healing time at final follow-up was 4.4 months in group A and 6 months in group B. Two (4.5%) cases of osteonecrosis of the femoral head occurred in group A, and 8 (23.5%) cases occurred in group B. Internal fixation with 2 cannulated compression screws combined with an iliac graft supported by the ascending branch of the lateral femoral circumflex artery is an effective surgical approach for treating femoral neck fractures in young adults to minimize the occurrence of fracture nonunion and osteonecrosis of the femoral head and to facilitate bone healing and functional recovery of the hip.