Multilayer three-dimensional super resolution imaging of thick biological samples

Recent advances in optical microscopy have enabled biological imaging beyond the diffraction limit at nanometer resolution. A general feature of most of the techniques based on photoactivated localization microscopy (PALM) or stochastic optical reconstruction microscopy (STORM) has been the use of thin biological samples in combination with total internal reflection, thus limiting the imaging depth to a fraction of an optical wavelength. However, to study whole cells or organelles that are typically up to 15 μm deep into the cell, the extension of these methods to a three-dimensional (3D) super resolution technique is required. Here, we report an advance in optical microscopy that enables imaging of protein distributions in cells with a lateral localization precision better than 50 nm at multiple imaging planes deep in biological samples. The approach is based on combining the lateral super resolution provided by PALM with two-photon temporal focusing that provides optical sectioning. We have generated super-resolution images over an axial range of ≈10 μm in both mitochondrially labeled fixed cells, and in the membranes of living S2 Drosophila cells.     

Gastrointestinal hormone abnormalities and G and D cells in functional dyspepsia patients with gastric dysmotility

AIM: To investigate the relationship between gastric dysmotility, gastrointestinal hormone abnormalities, and neuroendocrine cells in gastrointestinal mucosa in patients with functional dyspepsia (FD). METHODS: Gastric emptying was assessed with solid radiopaque markers in 54 FD patients, and the patients were divided into two groups according to the results, one with delayed gastric emptying and the other with normal gastric emptying. Seventeen healthy volunteers acted as normal controls. Fasting and postprandial plasma levels and gastroduodenal mucosal levels of gastrointestinal hormones gastrin, somatostatin (SS) and neurotensin (NT) were measured by radioimmunoassay in all the subjects. G cells (gastrin-producing cells) and D cells (SS-producing cells) in gastric antral mucosa were immunostained with rabbit anti-gastrin polyclonal antibody and rabbit anti-SS polyclonal antibody, respectively, and analyzed quantitatively by computerized image analysis. RESULTS: The postprandial plasma gastrin levels, the fasting and postprandial plasma levels and the gastric and duodenal mucosal levels of NT were significantly higher in the FD patients with delayed gastric emptying than in those with normal gastric emptying and normal controls. The number and gray value of G and D cells and the G cell/D cell number ratio did not differ significantly between normal controls and the FD patients with or without delayed gastric emptying. CONCLUSION: Our findings suggest that the abnormalities of gastrin and NT may play a role in the pathophysiology of gastric dysmotility in FD patients, and the abnormality of postprandial plasma gastrin levels in FD patients with delayed gastric emptying is not related to the changes both in the number and gray value of G cells and in the G cell/D cell number ratio in gastric antral mucosa.     

家庭-社会支持对3D打印辅助治疗复杂创伤骨折患者心理应激的影响

目的 探讨家庭-社会支持对3D打印辅助个体化定制治疗复杂创伤骨折患者心理应激的影响。 方法 以2015年1月-2017年2月我院收治的80例复杂创伤骨折患者为对象,按随机数字表法将其分为对照组和观察组各40例,2组均接受3D打印辅助个体化定制治疗,对照组实施认知行为干预,观察组在对照组基础上给予家庭-社会支持,比较干预前后2组汉密尔顿抑郁量表(Hamilton depression scale, HAMD)及汉密尔顿焦虑量表(Hamilton anxiety scale, HAMA)评分、视觉模拟疼痛(visual analogue scale/score, VAS)评分及生活质量调查问卷(the EORTC quality of life questionnaire-core 30,EORTC QLQ-C30)评分。 结果 干预后,2组HAMD、HAMA、VAS评分均低于对照组, EORTC QLQ-C30评分高于对照组。 结论 家庭-社会支持应用于3D打印辅助治疗复杂创伤骨折患者中,在改善其心理应激、术后疼痛和预后等方面具有积极作用,在临床中有推广应用价值。     

Efficacy of Injected Corticosteroid Type,Dose, and Volume for Pain in Large Joints: A Narrative Review

Corticosteroid injections are commonly used in the treatment of disorders of the large joints. This review assimilates the available literature on corticosteroid injections into the glenohumeral joint, subacromial space, hip joint, and knee joint. A systematic method to review available literature revealed 84 articles that fit the inclusion criteria. For each injection location, four items were examined: overall efficacy of injection, superior type of steroid, superior dose of steroid, and superior volume of injectate. Most research demonstrates positive short-term outcomes in pain and function for corticosteroid injections of the large joints. Methylprednisolone and triamcinolone seem similar in efficacy, with minor differences seen in specific studies. Larger doses may last longer, but need to be balanced with the systemic effects from higher doses. Volume has not been studied extensively. Due to heterogeneity in study types, subject populations, and outcomes, it is not possible to identify a single defining trend for a superior type, dose, or volume of steroid. Future prospective studies examining these factors may better reveal the optimum regimen for each injection location.

The Role of Rehabilitation in Patients With Progressive Supranuclear Palsy: A Narrative Review

Progressive supranuclear palsy (PSP) is a progressive neurodegenerative disorder caused by the deposition of abnormal proteins in neurons of the basal ganglia that limit motor ability, resulting in disability and reduced quality of life. So far, no pharmacologic therapy has been developed, and the treatment remains symptomatic. The aim of the present study is to perform a systematic investigation of the literature, and to determine the types and effects of rehabilitative interventions used for PSP. A search of all studies was conducted in MEDLINE/PubMed, the Cochrane Central Register of Controlled Trials, CINAHL, and EMBASE. Twelve studies were identified, including 6 case reports, 3 case series, one case-control study, one quasi?randomized trial (i.e. not truly random) with crossover design, and one randomized controlled trial, with 88 patients investigated overall. Rehabilitative interventions varied in type, number, frequency, and duration of sessions. The most commonly used clinical measures were the Progressive Supranuclear Palsy Rating Scale (PSPRS) and Unified Parkinson's Disease Rating Scale (UPDRS). Physical exercises were the main rehabilitative strategy but were associated with other interventions and rehabilitative devices, in particular treadmill and robot-assisted gait training. All studies showed an improvement in balance and gait impairment with a reduction of falls after rehabilitation treatment. Because of poor methodological quality and the variety of rehabilitative approaches including different and variable strategies, there was insufficient evidence of the effectiveness of any specific rehabilitation intervention in PSP. Despite this finding, rehabilitation might improve balance and gait, thereby reducing falls in PSP patients.

Benefits of Hippotherapy and Horse Riding Simulation Exercise on Healthy Older Adults: A Systematic Review

Objective

To provide an up-to-date research analysis on equine-assisted therapies and horse riding simulation exercise in older adults, and to suggest future directions in clinical practice and research.

Meniscus ECM‐functionalised hydrogels containing infrapatellar fat pad‐derived stem cells for bioprinting of regionally defined meniscal tissue

Injuries to the meniscus of the knee commonly lead to osteoarthritis. Current therapies for meniscus regeneration, including meniscectomies and scaffold implantation, fail to achieve complete functional regeneration of the tissue. This has led to increased interest in cell and gene therapies and tissue engineering approaches to meniscus regeneration. The implantation of a biomimetic implant, incorporating cells, growth factors, and extracellular matrix (ECM)‐derived proteins, represents a promising approach to functional meniscus regeneration. The objective of this study was to develop a range of ECM‐functionalised bioinks suitable for 3D bioprinting of meniscal tissue. To this end, alginate hydrogels were functionalised with ECM derived from the inner and outer regions of the meniscus and loaded with infrapatellar fat pad‐derived stem cells. In the absence of exogenously supplied growth factors, inner meniscus ECM promoted chondrogenesis of fat pad‐derived stem cells, whereas outer meniscus ECM promoted a more elongated cell morphology and the development of a more fibroblastic phenotype. With exogenous growth factors supplementation, a more fibrogenic phenotype was observed in outer ECM‐functionalised hydrogels supplemented with connective tissue growth factor, whereas inner ECM‐functionalised hydrogels supplemented with TGFβ3 supported the highest levels of Sox‐9 and type II collagen gene expression and sulfated glycosaminoglycans (sGAG) deposition. The final phase of the study demonstrated the printability of these ECM‐functionalised hydrogels, demonstrating that their codeposition with polycaprolactone microfibres dramatically improved the mechanical properties of the 3D bioprinted constructs with no noticeable loss in cell viability. These bioprinted constructs represent an exciting new approach to tissue engineering of functional meniscal grafts.     

Bone regeneration by means of a three‐dimensional printed scaffold in a rat cranial defect

Recently, computer‐designed three‐dimensional (3D) printing techniques have emerged as an active research area with almost unlimited possibilities. In this study, we used a computer‐designed 3D scaffold to drive new bone formation in a bone defect. Poly‐L‐lactide (PLLA) and bioactive β‐tricalcium phosphate (TCP) were simply mixed to prepare ink. PLLA + TCP showed good printability from the micronozzle and solidification within few seconds, indicating that it was indeed printable ink for layer‐by‐layer printing. In the images, TCP on the surface of (and/or inside) PLLA in the printed PLLA + TCP scaffold looked dispersed. MG‐63 cells (human osteoblastoma) adhered to and proliferated well on the printed PLLA + TCP scaffold. To assess new bone formation in vivo, the printed PLLA + TCP scaffold was implanted into a full‐thickness cranial bone defect in rats. The new bone formation was monitored by microcomputed tomography and histological analysis of the in vivo PLLA + TCP scaffold with or without MG‐63 cells. The bone defect was gradually spontaneously replaced with new bone tissues when we used both bioactive TCP and MG‐63 cells in the PLLA scaffold. Bone formation driven by the PLLA + TCP30 scaffold with MG‐63 cells was significantly greater than that in other experimental groups. Furthermore, the PLLA + TCP scaffold gradually degraded and matched well the extent of the gradual new bone formation on microcomputed tomography. In conclusion, the printed PLLA + TCP scaffold effectively supports new bone formation in a cranial bone defect.