miR-30a-5p Inhibits Proliferation and Migration of Lung Squamous Cell Carcinoma Cells by Targeting FOXD1.

OBJECTIVE: To investigate the mechanism of miR-30a-5p inhibiting proliferation and migration of lung squamous cell carcinoma (LSCC) cells by targeting FOXD1. METHODS: Bioinformatics was used to analyze differentially expressed genes in the TCGA_LUSC database. qRT-PCR was used to detect the expression levels of miR-30a-5p and FOXD1 in human normal lung epithelial cell line and human LSCC cell lines. The protein expression of FOXD1 was detected by western blot. The cell viability and colony formation abilities were examined by CCK-8 and colony formation assays, respectively. Wound healing and Transwell assays were performed to examine the migration and invasion abilities of cells. The targeted binding sites of miR-30a-5p and FOXD1 were predicted by bioinformatics, and dual luciferase assay was used to verify the targeted binding relationship between miR-30a-5p and FOXD1. RESULT: miR-30a-5p was downregulated in LSCC tissues and cells, while FOXD1 was highly expressed. Overexpression of miR-30a-5p or silencing FOXD1 inhibited cell viability, colony formation ability, migration, and invasion of LSCC cells. miR-30a-5p inhibited the proliferation and migration of LSCC cells by downregulating the expression of FOXD1. CONCLUSION: miR-30a-5p can downregulate the expression of FOXD1 and inhibit the proliferation and migration of LSCC.

Comparison of serum PCT and CRP levels in patients infected by different pathogenic microorganisms: a systematic review and meta-analysis.

To avoid the abuse and misuse of antibiotics, procalcitonin (PCT) and C-reactive protein (CRP) have been used as new approaches to identify different types of infection. Multiple databases were adopted to search relevant studies, and the articles that satisfied the inclusion criteria were included. Meta-analyses were conducted with Review Manager 5.0, and to estimate the quality of each article, risk of bias was assessed. Eight articles satisfied the inclusion criteria. The concentrations of both PCT and CRP in patients with bacterial infection were higher than those with non-bacterial infection. Both PCT and CRP levels in patients with G- bacterial infection were higher than in those with G+ bacterial infection and fungus infection. In the G+ bacterial infection group, a higher concentration of CRP was observed compared with fungus infection group, while the difference of PCT between G+ bacterial infection and fungus infection was not significant. Our study suggested that both PCT and CRP are helpful to a certain extent in detecting pneumonia caused by different types of infection.

Comparison of serum PCT and CRP levels in patients infected by different pathogenic microorganisms: a systematic review and meta-analysis

To avoid the abuse and misuse of antibiotics, procalcitonin (PCT) and C-reactive protein (CRP) have been used as new approaches to identify different types of infection. Multiple databases were adopted to search relevant studies, and the articles that satisfied the inclusion criteria were included. Meta-analyses were conducted with Review Manager 5.0, and to estimate the quality of each article, risk of bias was assessed. Eight articles satisfied the inclusion criteria. The concentrations of both PCT and CRP in patients with bacterial infection were higher than those with non-bacterial infection. Both PCT and CRP levels in patients with G− bacterial infection were higher than in those with G+ bacterial infection and fungus infection. In the G+ bacterial infection group, a higher concentration of CRP was observed compared with fungus infection group, while the difference of PCT between G+ bacterial infection and fungus infection was not significant. Our study suggested that both PCT and CRP are helpful to a certain extent in detecting pneumonia caused by different types of infection.

Spatially heterogeneous corneal mechanical responses before and after riboflavin-ultraviolet-A crosslinking.

PURPOSE: To determine the heterogeneous through-thickness strains in the cornea at physiologic intraocular pressures before and after corneal collagen crosslinking (CXL) using noninvasive ultrasound. SETTING: Department of Biomedical Engineering, Ohio State University, Columbus, Ohio, USA. DESIGN: Experimental study. METHODS: Sixteen paired canine corneoscleral shells were divided into 2 groups. The CXL group completed a standard CXL protocol using riboflavin-ultraviolet-A (UVA) irradiation. The control group was given an identical treatment except UVA irradiation. Ultrasound scans (at 55 MHz) of the cornea were obtained before and after treatment as the corneoscleral shell was inflated from 5 mm Hg to 45 mm Hg to calculate the distributive through-thickness strains in the cornea. The mean radial and tangential strains of the whole cornea layer, as well as those of the anterior, middle, and posterior thirds of the cornea, were compared before and after treatment in the control group and CXL group using linear mixed models with repeated measures. RESULTS: Significant reductions in tangential and radial strains occurred in the CXL group (P=.003 and P=.0025, respectively) but not the control group (P=.08 and P=.63, respectively). The anterior third had the smallest strains in all pretreated corneas (P<.001) and posttreated corneas (CXL group, P=.023; control group, P=.01). CONCLUSIONS: Ultrasound speckle tracking showed heterogeneous strain distributions through the cornea and confirmed that CXL results in a stiffer corneal response (ie, smaller strains during physiologic loadings). This technique may provide a clinical tool to quantify the biomechanical effects of CXL. FINANCIAL DISCLOSURE: No author has a financial or proprietary interest in any material or method mentioned.

Biaxial mechanical testing of posterior sclera using high-resolution ultrasound speckle tracking for strain measurements.

This study aimed to characterize the mechanical responses of the sclera, the white outer coat of the eye, under equal-biaxial loading with unrestricted shear. An ultrasound speckle tracking technique was used to measure tissue deformation through sample thickness, expanding the capabilities of surface strain techniques. Eight porcine scleral samples were tested within 72 h postmortem. High resolution ultrasound scans of scleral cross-sections along the two loading axes were acquired at 25 consecutive biaxial load levels. An additional repeat of the biaxial loading cycle was performed to measure a third normal strain emulating a strain gage rosette for calculating the in-plane shear. The repeatability of the strain measurements during identical biaxial ramps was evaluated. A correlation-based ultrasound speckle tracking algorithm was used to compute the displacement field and determine the distributive strains in the sample cross-sections. A Fung type constitutive model including a shear term was used to determine the material constants of each individual specimen by fitting the model parameters to the experimental stress-strain data. A non-linear stress-strain response was observed in all samples. The meridian direction had significantly larger strains than that of the circumferential direction during equal-biaxial loadings (P's<0.05). The stiffness along the two directions was also significantly different (P=0.02) but highly correlated (R(2)=0.8). These results showed that the mechanical properties of the porcine sclera were nonlinear and anisotropic under biaxial loading. This work has also demonstrated the feasibility of using ultrasound speckle tracking for strain measurements during mechanical testing.

Correlation between biomechanical responses of posterior sclera and IOP elevations during micro intraocular volume change.

PURPOSE: This study tested the hypothesis that intraocular pressure (IOP) elevations, induced by controlled increase of intraocular volume, are correlated with the biomechanical responses of the posterior sclera. METHODS: Porcine globes were tested within 48 hours postmortem. The first group of globes (n = 11) was infused with 15 µL of phosphate-buffered saline at three different rates to investigate rate-dependent IOP elevations. The second group (n = 16) was first infused at the fast rate and then underwent inflation tests to investigate the relationship between IOP elevations (ΔIOP) and scleral strains. The strains in the superotemporal region of the posterior sclera were measured by ultrasound speckle tracking. Linear regression was used to examine the association between ΔIOP due to micro-volumetric infusion and the scleral strains at a specific inflation pressure. RESULTS: The average ΔIOP was 14.9 ± 4.3 mm Hg for the infusion of 15 µL in 1 second. The ΔIOP was greater for the faster infusion rates but highly correlated across different rates (P < 0.001). A significant negative association was found between the ΔIOP and the tangential strains in both the circumferential (R(2) = 0.54, P = 0.003) and meridian (R(2) = 0.53, P = 0.002) directions in the posterior sclera. CONCLUSIONS: This study showed a substantial increase in IOP, with a large intersubject variance during micro-volumetric change. A stiffer response of the sclera was associated with larger IOP spikes, providing experimental evidence linking corneoscleral biomechanics to IOP fluctuation. In vivo measurement of corneoscleral biomechanics may help better predict the dynamic profile of IOP.

Experimental test of escape theory: accessibility to implicit suicidal mind.

This study tested the Escape Theory prediction that individuals blaming themselves for failure experience increased accessibility to implicit suicidal mind. One hundred and thirty-eight undergraduate medical students were randomly assigned to three groups: failure-related priming, success-related priming, and control. Following experimental conditions, participants completed a death/suicide Implicit Association Test. Results revealed significant differences between groups in accessibility to implicit suicidal mind. Furthermore, priming manipulation interacted with individual differences in locus of control (LOC). Significant differences in accessibility to implicit suicidal mind were observed in individuals with internal LOC, while effects of priming manipulation were eliminated in individuals with external LOC.

Ultrasonic measurement of scleral cross-sectional strains during elevations of intraocular pressure: method validation and initial results in posterior porcine sclera.

BACKGROUND: Scleral biomechanical properties may be important in the pathogenesis and progression of glaucoma. The goal of this study is to develop and validate an ultrasound method for measuring cross-sectional distributive strains in the sclera during elevations of intraocular pressure (IOP). METHOD OF APPROACH: Porcine globes (n = 5) were tested within 24 hs postmortem. The posterior scleral shells were dissected and mounted onto a custom-built pressurization chamber. A high-frequency (55-MHz) ultrasound system (Vevo660, VisualSonics Inc., Toronto) was employed to acquire the radio frequency data during scans of the posterior pole along both circumferential and meridian directions. The IOP was gradually increased from 5 to 45 mmHg. The displacement fields were obtained from correlation-based ultrasound speckle tracking. A least-square strain estimator was used to calculate the strains in both axial and lateral directions. Experimental validation was performed by comparing tissue displacements calculated from ultrasound speckle tracking with those induced by an actuator. Theoretical analysis and simulation experiments were performed to optimize the ultrasound speckle tracking method and evaluate the accuracy and signal-to-noise ratio (SNR) in strain estimation. RESULTS: Porcine sclera exhibited significantly larger axial strains (e.g., -5.1 ± 1.5% at 45 mmHg, meridian direction) than lateral strains (e.g., 2.2 ± 0.7% at 45 mmHg, meridian direction) during IOP elevations (P's < 0.01). The strain magnitudes increased nonlinearly with pressure increase. The strain maps displayed heterogeneity through the thickness. The lateral strains were significantly smaller in the circumferential direction than the meridian direction at 45 mmHg (P < 0.05). Experimental validation showed that the ultrasound speckle tracking method was capable of tracking displacements at the accuracy of sub-micron to micron. Theoretical analysis predicted the dependence of the strain estimation SNR on the strain level, as well as signal processing parameters such as kernel size. Simulation results showed that ultrasound speckle tracking had a high accuracy for estimating strains of 1-5% and a high SNR for strains of 0.5-5%. CONCLUSIONS: A new experimental method based on ultrasound speckle tracking has been developed for obtaining cross-sectional strain maps of the posterior sclera. This method provides a useful tool to examine distributive strains through the thickness of the sclera during elevations of IOP.

Vision-guided ocular growth in a mutant chicken model with diminished visual acuity.

Visual experience is known to guide ocular growth. We tested the hypothesis that vision-guided ocular growth is disrupted in a model system with diminished visual acuity. We examine whether ocular elongation is influenced by form-deprivation (FD) and lens-imposed defocus in the Retinopathy, Globe Enlarged (RGE) chicken. Young RGE chicks have poor visual acuity, without significant retinal pathology, resulting from a mutation in guanine nucleotide-binding protein ß3 (GNB3), also known as transducin ß3 or Gß3. The mutation in GNB3 destabilizes the protein and causes a loss of Gß3 from photoreceptors and ON-bipolar cells (Ritchey et al., 2010). FD increased ocular elongation in RGE eyes in a manner similar to that seen in wild-type (WT) eyes. By comparison, the excessive ocular elongation that results from hyperopic defocus was increased, whereas myopic defocus failed to significantly decrease ocular elongation in RGE eyes. Brief daily periods of unrestricted vision interrupting FD prevented ocular elongation in RGE chicks in a manner similar to that seen in WT chicks. Glucagonergic amacrine cells differentially expressed the immediate early gene Egr1 in response to growth-guiding stimuli in RGE retinas, but the defocus-dependent up-regulation of Egr1 was lesser in RGE retinas compared to that of WT retinas. We conclude that high visual acuity, and the retinal signaling mediated by Gß3, is not required for emmetropization and the excessive ocular elongation caused by FD and hyperopic defocus. However, the loss of acuity and Gß3 from RGE retinas causes enhanced responses to hyperopic defocus and diminished responses to myopic defocus.

The combination of IGF1 and FGF2 and the induction of excessive ocular growth and extreme myopia.

Different growth factors have been shown to influence the development of form-deprivation myopia and lens-induced ametropias. However, growth factors have relatively little effect on the growth of eyes with unrestricted vision. We investigate whether the combination of insulin-like growth factor 1 (IGF1) and fibroblast growth factor 2 (FGF2) influence ocular growth in eyes with unrestricted vision. Different doses of IGF1 and FGF2 were injected into the vitreous chamber of postnatal chicks. Measurements of ocular dimensions and intraocular pressure (IOP) were made during and at the completion of different treatment paradigms. Histological and immunocytochemical analyses were performed to assess cell death, cellular proliferation and integrity of ocular tissues. Treated eyes had significant increases in equatorial diameter and vitreous chamber depth. With significant variability between individuals, IGF1/FGF2-treatment caused hypertrophy of lens and ciliary epithelia, lens thickness was increased, and anterior chamber depth was decreased. Treated eyes developed myopia, in excess of 15 diopters of refractive error. Shortly after treatment, eyes had increased intraocular pressure (IOP), which was increased in a dose-dependent manner. Seven days after treatment with IGF1 and FGF2 changes to anterior chamber depth, lens thickness and elevated IOP were reduced, whereas increases in the vitreous chamber were persistent. Some damage to ganglion cells was detected in peripheral regions of the retina at 7 days after treatment. We conclude that the extreme myopia in IGF1/FGF2-treated eyes results from increased vitreous chamber depth, decreased anterior chamber depth, and changes in the lens. We propose that factor-induced ocular enlargement and myopia result from changes to the sclera, lens and anterior chamber depth.

Effect of corneal stiffening on Goldmann applanation tonometry and Tono-Pen measurements in canine eyes.

PURPOSE: To experimentally examine the effect of increased corneal stiffness on Goldmann applanation tonometry (GAT) and Tono-Pen (Reichert, Inc., Depew, NY) measurements of intraocular pressure (IOP) in a canine eye model. METHODS: Twenty globes were recovered from 10 dogs with no known diseases. For each dog, corneal stiffening was induced in one eye with glutaraldehyde/phosphate buffered saline (PBS) immersion while the other cornea was immersed in PBS only. Acoustic impedance was measured before and after treatment in all eyes. After treatment, IOP was measured by GAT and Tono-Pen at true pressures of 10, 15, 20, 30, and 40 mm Hg. The corneas were then dissected for uniaxial tensile testing. The GAT/Tono-Pen readings, corneal stiffness (measured by ultrasound and tensile tests), and corneal thickness were compared between the two groups. The correlations between GAT/Tono-Pen readings and corneal stiffness were evaluated. RESULTS: Acoustic impedance significantly increased after glutaraldehyde treatment (P < 0.01). Secant modulus at 1% strain was significantly higher in corneas treated with glutaradehyde/PBS than those treated with PBS only (P < 0.01). GAT and Tono-Pen readings were significantly higher at all pressure levels (P < 0.001) in the eyes with corneal stiffening. Both corneal acoustic impedance and secant modulus were significantly correlated with GAT/Tono-Pen readings at all pressure levels (P < 0.01). CONCLUSIONS: This study provided experimental evidence that corneal stiffening significantly increases GAT and Tono-Pen readings in canine eyes. Noninvasive ultrasound measurement of acoustic impedance may be used to evaluate corneal stiffness and improve the accuracy of clinical measurements of IOP.

Corneal modulus and IOP measurements in canine eyes using Goldmann applanation tonometry and Tono-pen.

PURPOSE: To experimentally examine the effect of corneal modulus on Goldmann applanation tonometry (GAT) and Tono-pen (Tono-pen XL, Reichert, Inc., Depew, NY) measurements of intraocular pressure (IOP) in a canine eye model. METHODS: Twenty-one canine globes were recovered from healthy animals. IOP was controlled at 10, 15, 20, 30, and 40 mm Hg and measured by GAT and Tono-pen following standard protocols. The corneas were dissected and uniaxial tensile tests were performed on corneal strips. The correlation between GAT and Tono-pen errors and corneal secant modulus was evaluated using Pearson correlation coefficients. The influence of corneal thickness and the true pressure was also examined. RESULTS: At a true IOP of 10, 15, 20, 30, and 40 mm Hg, the GAT readings were 1.1 ± 1.0, 5.1 ± 1.5, 9.5 ± 2.0, 17.3 ± 1.6, and 25.3 ± 1.8 mm Hg, respectively. The corresponding Tono-pen readings were 7.8 ± 1.7, 12.4 ± 1.7, 16.1 ± 1.9, 22.5 ± 2.1, and 28.1 ± 2.2 mm Hg, respectively. The mean secant modulus at 1% strain of the canine corneal strips was 1.54 ± 0.43 megapascal (MPa). Corneal secant modulus was significantly correlated with GAT errors when the true IOP was 30 mm Hg (R = 0.49; P < 0.05). No significant correlation was observed between tonometric errors and corneal thickness. Both GAT and Tono-pen errors increased significantly at higher pressures (P < 0.001). CONCLUSIONS: Both GAT and Tono-pen underestimated IOP in canine eyes. There was preliminary experimental evidence for a correlation between corneal modulus and GAT in the canine eyes and a higher corneal modulus was associated with higher GAT readings at a certain pressure level. The tonometric errors appeared to be pressure-dependent.

Variance of speed of sound and correlation with acoustic impedance in canine corneas.

The clinical standard for measuring corneal thickness is ultrasound pachymetry that assumes a constant speed of sound. The purpose of this study was to examine the variance of speed of sound and its relationship with acoustic impedance in healthy eyes of canines with a large age span. Corneal speed of sound and acoustic impedance were measured in 34 canine eyes at room temperature (21 ± 1°C). The mean speed of sound was 1577 ± 10 m/s ranging from 1553 to 1594 m/s. There was a strong correlation between speed of sound and acoustic impedance (R = 0.84, p < 0.001). Corneal speed of sound had a small variance in healthy canines over 1-year-old, but was significantly lower in younger canines suggesting an age effect. The strong correlation between corneal speed of sound and acoustic impedance may offer a potential means to noninvasively detect abnormal speed of sound for more accurate corneal thickness estimation.

Measurement of corneal changes after collagen crosslinking using a noninvasive ultrasound system.

PURPOSE: To determine the changes in corneal properties induced by riboflavin-ultraviolet A (UVA) irradiation using a noninvasive ultrasound (US) method in a porcine eye model. SETTING: Department of Biomedical Engineering, Ohio State University, Columbus, Ohio, USA. METHODS: Fresh porcine globes acquired within 24 hours postmortem were divided into equal groups. In the collagen crosslinking (CXL) group, the globes were irradiated with UVA in combination with riboflavin application following the standard protocol. The control group had the same procedures as the CXL group but with no UVA irradiation. Corneal properties were measured before and after treatments in both groups using a quantitative US method. Changes in corneal thickness, density, and stiffness were examined and the difference between the changes in the CXL group and the control group was analyzed. RESULTS: Each group comprised 14 porcine globes. After riboflavin-UVA treatment, there was a significant increase in corneal stiffness (mean 2.59 +/- 0.08 GPa before and 2.70 +/- 0.08 GPa after CXL) (P<.001) and in the speed of sound in corneal tissue (1557 +/- 23 m/s versus 1588 +/- 22 m/s) (P<.001). There was no significant change in corneal stiffness or in the speed of sound in the control group. Corneal thickness increased significantly after the treatment in both groups. No significant change in corneal density was observed in either group. CONCLUSION: Corneal stiffness increased after riboflavin-UVA CXL, a finding detected using a noninvasive US technique. FINANCIAL DISCLOSURE: Dr. Liu has filed a patent application for the general ultrasound technique used in the study. No other author has a financial or proprietary interest in any material or method mentioned.