Integrative Analysis of miRNA and circRNA Expression Profiles and Interaction Network in HSV-1-Infected Primary Corneal Epithelial Cells.

PURPOSE: Circular RNAs (circRNAs) are products of alternative splicing with roles as competitive endogenous RNAs or microRNA sponges, regulating gene expression and biological processes. However, the involvement of circRNAs in herpes simplex keratitis remains largely unexplored. METHODS: This study examines circRNA and miRNA expression profiles in primary human corneal epithelial cells infected with HSV-1, compared to uninfected controls, using microarray analysis. Bioinformatic analysis predicted the potential function of the dysregulated circRNAs and microRNA response elements (MREs) in these circRNAs, forming an interaction network between dysregulated circRNAs and miRNAs. RESULTS: A total of 332 circRNAs and 16 miRNAs were upregulated, while 80 circRNAs and six miRNAs were downregulated (fold change ≥2.0 and p < 0.05). Gene ontology (GO) and KEGG pathway analyses were performed on parental genes of dysregulated circRNAs to uncover potential functions in HSV-1 infection. Notably, miR-181b-5p, miR-338-3p, miR-635, and miR-222-3p emerged as pivotal miRNAs interacting with multiple dysregulated circRNAs. CONCLUSIONS: This comprehensive study offers insights into differentially expressed circRNAs and miRNAs during HSV-1 infection in corneal epithelial cells, shedding light on circRNA-miRNA interactions' potential role in herpes simplex keratitis pathogenesis.

Acute effect of a variable pulse width Nd:YAG laser combined with hematoporphyrin monomethyl ether-mediated photodynamic therapy on a cockscomb model of nevus flammeus.

BACKGROUND: Nevus flammeus (NF) is a congenital vascular malformation. OBJECTIVES: To investigate the acute effect of a variable pulse width Nd:YAG laser combined with hematoporphyrin monomethyl ether (HMME)-mediated photodynamic therapy (PDT) on a cockscomb model of NF. MATERIAL AND METHODS: Forty-two leghorn roosters were randomly divided into the following 7 groups: group A1 (treated with HMME-mediated PDT; energy density of 75 J/cm2), group A2 (treated with HMME-mediated PDT; 125 J/cm2), group A3 (treated with HMME-mediated PDT; 150 J/cm2), group A4 (treated with HMME-mediated PDT; 175 J/cm2), group B (treated with a variable pulse width Nd:YAG laser), group C (treated with a variable pulse width Nd:YAG laser and HMME-mediated PDT), and group D (the control group). Changes in the cockscomb tissues were observed visually and microscopically on days 1, 3, 7, and 14 after treatment. The capillary reduction and the ratio of collagen type I to type III were examined. RESULTS: The response rate was higher in groups A3 and A4 than in group B. In group A, a higher energy density resulted in a higher response rate and a greater capillary reduction (p < 0.05 for all). However, we concluded that PDT at an energy density of 175 J/cm2 is not suitable for treating NF, as severe tissue damage, markedly lower capillary numbers, and markedly higher collagen type I:III ratios were observed in the cockscombs treated at this energy density; instead, 150 J/cm2 may be a more appropriate energy density. Moreover, HMME-mediated PDT at 150 J/cm2 combined with a variable pulse width Nd:YAG laser achieved better treatment outcomes than PDT or a variable pulse width Nd:YAG laser alone (p < 0.05 for both). CONCLUSIONS: Compared to PDT or a variable pulse width Nd:YAG laser alone, the combination of the 2 therapies achieved a better acute effect in treating a cockscomb model of NF, and 150 J/cm2 may be a suitable energy density for PDT.

Premorbid Alterations of Spontaneous Brain Activity in Elderly Patients With Early Post-operative Cognitive Dysfunction: A Pilot Resting-State Functional MRI Study.

Background: Elderly patients with pre-existing cognitive impairment are susceptible to post-operative cognitive dysfunction (POCD). In this study, we investigated whether there is pre-existing local homogeneity and functional connectivity alteration in the brain before surgery for POCD patients as compared to that in non-POCD patients. Methods: Eighty elderly patients undergoing major thoracic or abdominal surgeries were recruited. Resting-state functional MRI was scanned at least 1 day before surgery. Neuropsychological tests (NPTs) were performed before surgery and at discharge, respectively. Pre-operative regional homogeneity (ReHo) and resting-state functional connectivity (RSFC) were compared between POCD patients and non-POCD patients, respectively. Partial correlation between NPTs and ReHo or RSFC was analyzed by adjusting for confounding factors. Results: Significant difference (P < 0.001, Gaussian Random Field (GRF) correction which is a multiple comparisons correction method at cluster level, cluster size > 49) in ReHo between POCD patients and non-POCD patients was detected in right hippocampus/parahippocampus. Pre-operative RSFC between right hippocampus/parahippocampus and right middle/inferior temporal gyrus increased in POCD patients (P < 0.001, GRF correction for multiple comparisons) when compared with that in non-POCD patients.RSFC significantly correlated with composite Z-score (r = 0.46, 95% CI [0.234, 0.767], P = 0.002) or Digit Symbol Substitution Test Z-scores (r = 0.31, 95% CI [0.068, 0.643], P = 0.046) after adjusting for confounding factors. Conclusions: The results suggest that premorbid alterations of spontaneous brain activity might exist in elderly patients who develop early POCD. The neural mechanism by which patients with pre-operative abnormal spontaneous activity are susceptible to POCD requires further study.

An improved cellular enucleation method with extracellular matrix and colchicine facilitates the study of nucleocytoplasmic interaction.

Enucleated mammalian cells (cytoplasts) have been widely used for studying differential roles of the cytoplasm and nucleus in various cellular processes. Here, we reported an improved enucleation protocol, in which cells were seeded in extracellular matrix (ECM)-coated 24-wells and spun at 4600 g and 35 °C for 60 min in the presence of cytochalasin B and colchicine. When glass-bottom wells were used, cellular structures and organelles in cytoplasts could be examined directly by confocal microscopy. Nuclear envelope rupture did not occur probably due to mild centrifugation conditions used in this study. Addition of paclitaxel or doxorubicin completely blocked proliferation of residual nucleated cells; however, to our surprise, paclitaxel dramatically prolonged the survival of cytoplasts. Results from Annexin V and Propidium Iodide staining showed that cytoplasts died predominantly by apoptosis, which was partially inhibited by ECM and further by paclitaxel. Mitochondria were mostly rod-shaped and formed a connected network in paclitaxel-treated cytoplasts, indicating lack of fusion and fission dynamics. Moreover, paclitaxel increased mitochondrial membrane potential, suggesting that perturbation of mitochondria might be critical to the survival of cytoplasts. In conclusion, we had established an efficient and fast procedure for enucleation of adherent animal cells, which could facilitate the investigation of nucleocytoplasmic interaction.

The effects of both age and sex on irisin levels in paired plasma and cerebrospinal fluid in healthy humans.

BACKGROUND: Adipo-myokine irisin has important effects on the metabolism and functioning of multiple tissues and organs. However, the effects of aging and sex on irisin levels in cerebrospinal fluid (CSF) and on circulation have not been comprehensively studied. OBJECTIVE: To investigate whether aging and sex can affect irisin levels in both CSF and plasma; to determine whether CSF irisin uptake involves a saturable transport mechanism. DESIGN AND METHODS: In the present study, the irisin levels in paired CSF and plasma samples drawn from 71 healthy individuals were used to investigate effects by using commercial ELISA kits and mass spectrometry. RESULTS: Multiple linear regression analysis results showed that CSF irisin levels are positively correlated with the CSF/plasma irisin ratio and age and that these levels present a reverse correlation with BMI. Age-related increases in CSF levels are validated by using ELISA and mass spectrometry. Higher plasma irisin levels are observed in men than women. CSF and plasma irisin levels are nonlinearly associated with the CSF/plasma irisin ratio, BMI, age and F scores. The CSF/plasma irisin ratio is U-shaped and associated with age. CONCLUSIONS: There might be an age-related increase in irisin levels in the cerebrospinal fluid of healthy humans. Circulating irisin levels are higher in males than in females in the healthy population. A saturable mechanism might be involved in mediating the transport of circulating irisin across the blood-brain barrier. Factors shaping irisin levels for both circulation and the CSF of healthy humans must be further defined in future experiments.

Efficiency of stem cell based therapy in the treatment of diabetic foot ulcer: a meta-analysis.

Diabetic foot ulcer is a chronic, refractory, frequent complication in diabetic patient. Its treatment often requires multidisciplinary joint efforts, diverse strategies have been adopted to address this annoying issue, including stem cell-based therapy/acellular dermal matrix/negative pressure wound therapy etc. However, consensus has not been reached. To assess the current evidence regarding the efficiency and potential advantages of stem cell-based therapy compared with conventional standard treatment and/or placebo in the treatment of diabetic foot ulcer. A comprehensive search in PubMed, EmBase, Cochrane Central and Web of Science databases was conducted during December 2016 and a systematic review and meta-analysis of all relevant studies were performed. A total of 7 studies that involved 224 diabetic foot patients, classified as Wagner grades 1-5, were analyzed. The pooled results confirmed the benefits of using the stem cell treatment. Partial and/or complete healing were significantly higher in the stem cell group compared with the control group (77.4% vs. 31.9%; RR: 2.22; 95% CI, 1.65-2.98). Subgroup analysis on ABI and TCP02 also confirmed the results. The present meta-analysis indicates that stem cell-based therapy can enhance the healing of diabetic foot ulcers and is associated with lesser pain, lower amputation rate and improved prognosis compared with normal treatment. Well-designed randomized controlled trials are required in the future in order to confirm and update these findings.

Comparison of the impact of prolonged low-pressure and standard-pressure pneumoperitoneum on myocardial injury after robot-assisted surgery in the Trendelenburg position: study protocol for a randomized controlled trial.

BACKGROUND: Robot-assisted laparoscopic radical prostatectomy and robot-assisted radical cystectomy have gradually become the preferred choices for urologists as they allow surgeons to perform complex procedures more precisely and effectively. The pneumoperitoneum, which is normally applied in these surgeries to provide visual clarity and space to perform the procedure, may cause hemodynamic disturbance, potentially myocardial injury. Thus surgeons have recently considered opting for the low-pressure pneumoperitoneum to lower this negative impact. Herein we describe a protocol for a clinical trial to compare the impact of prolonged low-pressure and standard-pressure pneumoperitoneum on myocardial injury after robot-assisted surgery. METHODS/DESIGN: This study is designed to be a bicenter clinical trial. In total 280 patients scheduled to undergo robot-assisted laparoscopic radical prostatectomy or robot-assisted radical cystectomy will be enrolled and randomized into two groups, with standard- (12-16 mmHg) and low-pressure (7-10 mmHg) pneumoperitoneum, respectively. Troponin T will be measured as the primary endpoint to assess the extent of myocardial injury. Nt-proBNP and hemodynamic indexes will also be recorded for further analysis. DISCUSSION: The significance of this study is emphasized by the fact that there are few studies that have focused on the impact of prolonged pneumoperitoneum on myocardial injury, which is relevant to postoperative mortality. We hope that the conclusions drawn from this study could provide reference and basis to the future of the pneumoperitoneum in clinical practice. TRIAL REGISTRATION: Registered at https://www.clinicaltrials.gov with the Identifier NCT02600481 on November 5, 2015.

Matrigel basement membrane matrix induces eccrine sweat gland cells to reconstitute sweat gland-like structures in nude mice.

BACKGROUND: Severe burn results in irreversible damage to eccrine sweat glands, for which no effective treatment is available. Interaction between the extracellular matrix and epithelial cells is critical for proper three-dimensional organization and function of the epithelium. METHODS: Matrigel-embedded eccrine sweat gland cells were subcutaneously implanted into the inguinal regions of nude mice. Two weeks later, the Matrigel plugs were removed and evaluated for series of detection items. RESULTS: Sweat gland cells developed into sweat gland-like structures in the Matrigel plugs based on: (1) de novo formation of tubular-like structures with one or more hollow lumens, (2) expression of epithelial and sweat gland markers (pancytokeratin, CK5/7/14/19, α-SMA and CEA), (3) basement membrane formation, (4) myoepithelial cells presenting in and encompassing the tubular-like structures, (5) cellular polarization, evident by the expression of tight junction proteins (claudin-1 and ZO-2), anchoring junctions (desmoglein-1 and -2 and E-cadherin) and CEA in the luminal membrane, (6) expression of proteins related to sweat secretion and absorption (Na(+)-K(+)-ATPase α/ß, Na(+)-K(+)-2Cl-cotranspoter 1, Na(+)/H(+) exchanger 1, aquaporin-5, epithelial sodium channel, cystic fibrosis transmembrane conductance regulator, potassium channel and vacuolar-type H+-ATPase), and (7) about 20% of the tubular-like structures are de novo coils and 80% are de novo ducts. CONCLUSIONS: This study provides not only an excellent model to study eccrine sweat gland development, cytodifferentiation and reconstitution, but also an in vivo model for regeneration of eccrine sweat glands.

Localization of Na(+)-K(+)-ATPase α/ß, Na(+)-K(+)-2Cl-cotransporter 1 and aquaporin-5 in human eccrine sweat glands.

In order to evaluate the function of the repaired or regenerated eccrine sweat glands, we must first localize the proteins involved in sweat secretion and absorption in normal human eccrine sweat glands. In our studies, the cellular localization of Na(+)-K(+)-ATPase α/ß, Na(+)-K(+)-2Cl-cotransporter 1 (NKCC1) and aquaporin-5 (AQP5) in eccrine sweat glands were detected by immunoperoxidase labeling. The results showed that Na(+)-K(+)-ATPase α was immunolocalized in the cell membrane of the basal layer and suprabasal layer cells of the epidermis, the basolateral membrane of the secretory coils, and the cell membrane of the outer cells and the basolateral membrane of the luminal cells of the ducts. The localization of Na(+)-K(+)-ATPase ß in the secretory coils was the same as Na(+)-K(+)-ATPase α, but Na(+)-K(+)-ATPase ß labeling was absent in the straight ducts and epidermis. NKCC1 labeling was seen only in the basolateral membrane of the secretory coils. AQP5 was strongly localized in the apical membrane and weakly localized in the cytoplasm of secretory epithelial cells. The different distribution of these proteins in eccrine sweat glands was related to their functions in sweat secretion and absorption.

The cellular localization of Na(+)/H(+) exchanger 1, cystic fibrosis transmembrane conductance regulator, potassium channel, epithelial sodium channel γ and vacuolar-type H+-ATPase in human eccrine sweat glands.

The secretory portions of human eccrine sweat glands secrete isotonic fluid into the lumen and then the primary fluid is rendered hypotonic during its passage to the skin surface. During the processes of sweat secretion and absorption, many enzymes and proteins play important roles. In the study, the cellular localizations of Na(+)/H(+) exchanger 1 (NHE1), cystic fibrosis transmembrane conductance regulator (CFTR), potassium channel (KC), epithelial sodium channel γ (γENaC) and vacuolar-type H+-ATPase (V-ATPase) in human eccrine sweat glands and epidermis were detected using immunofluorescence labeling. The results revealed that in the secretory coils, the basolateral membranes showed evidence of CFTR, NHE1 and KC activities, the apical membranes showed the activities of KC and NHE1, and the nucleus showed γEaNC and V-ATPase activities; in the duct, the peripheral and luminal ductal cells showed evidence of CFTR, NHE1 and KC, the apical membranes showed the activities of CFTR and NHE1, and the nucleus showed γEaNC, V-ATPase and KC activities. The cellular localization of these proteins in eccrine sweat glands is helpful to better understand the mechanisms of sweat secretion and absorption.