Yi-Hsiung Lee

Nodal promotes growth and invasion in human gliomas.

Uncontrolled growth and diffused invasion are major causes of mortality in patients with malignant gliomas. Nodal has been shown to have a central role in the tumorigenic signaling pathways of malignant melanoma. In this study, we show that grade IV human glioma cell lines expressed different levels of Nodal, paralleled to the potential for cell invasiveness. Treatment of glioma cell lines with recombinant Nodal (rNodal) increased matrix metalloproteinase 2 (MMP-2) secretion and cell invasiveness. The ectopic expression of Nodal in GBM glioma cells that expressed Nodal at low level resulted in increased MMP-2 secretion, enhanced cell invasiveness, raised cell proliferation rates in vitro, increased tumor growth in vivo, and was associated with poor survival in a mice xenograft model. In contrast, the knockdown of Nodal expression in U87MG glioma cells with high Nodal expression level had reduced MMP-2 secretion, less cell invasiveness, lower tumor growth in vivo and longer lifespan in mice with U87MG/shNodal cell xenografts. In addition, Nodal knockdown promoted the reversion of malignant glioma cells toward a differentiated astrocytic phenotype. Furthermore, our data support the notion that Nodal may regulate glioma progression through the induction of the leukemia inhibitory factor (LIF) and Cripto-1 through activated Smad.

Hematoma Shape, Hematoma Size, Glasgow Coma Scale Score and ICH Score: Which Predicts the 30-Day Mortality Better for Intracerebral Hematoma?

Purpose To investigate the performance of hematoma shape, hematoma size, Glasgow coma scale (GCS) score, and intracerebral hematoma (ICH) score in predicting the 30-day mortality for ICH patients. To examine the influence of the estimation error of hematoma size on the prediction of 30-day mortality. Materials and Methods This retrospective study, approved by a local institutional review board with written informed consent waived, recruited 106 patients diagnosed as ICH by non-enhanced computed tomography study. The hemorrhagic shape, hematoma size measured by computer-assisted volumetric analysis (CAVA) and estimated by ABC/2 formula, ICH score and GCS score was examined. The predicting performance of 30-day mortality of the aforementioned variables was evaluated. Statistical analysis was performed using Kolmogorov-Smirnov tests, paired t test, nonparametric test, linear regression analysis, and binary logistic regression. The receiver operating characteristics curves were plotted and areas under curve (AUC) were calculated for 30-day mortality. A P value less than 0.05 was considered as statistically significant. Results The overall 30-day mortality rate was 15.1% of ICH patients. The hematoma shape, hematoma size, ICH score, and GCS score all significantly predict the 30-day mortality for ICH patients, with an AUC of 0.692 (P = 0.0018), 0.715 (P = 0.0008) (by ABC/2) to 0.738 (P = 0.0002) (by CAVA), 0.877 (P<0.0001) (by ABC/2) to 0.882 (P<0.0001) (by CAVA), and 0.912 (P<0.0001), respectively. Conclusion Our study shows that hematoma shape, hematoma size, ICH scores and GCS score all significantly predict the 30-day mortality in an increasing order of AUC. The effect of overestimation of hematoma size by ABC/2 formula in predicting the 30-day mortality could be remedied by using ICH score.

Multiplying optical tweezers force using a micro-lever.

This study presents a photo-driven micro-lever fabricated to multiply optical forces using the two-photon polymerization 3D-microfabrication technique. The micro-lever is a second class lever comprising an optical trapping sphere, a beam, and a pivot. A micro-spring is placed between the short and long arms to characterize the induced force. This design enables precise manipulation of the micro-lever by optical tweezers at the micron scale. Under optical dragging, the sphere placed on the lever beam moves, resulting in torque that induces related force on the spring. The optical force applied at the sphere is approximately 100 to 300 pN, with a laser power of 100 to 300 mW. In this study, the optical tweezers drives the micro-lever successfully. The relationship between the optical force and the spring constant can be determined by using the principle of leverage. The arm ratio design developed in this study multiplies the applied optical force by 9. The experimental results are in good agreement with the simulation of spring property.

The cerebral vasomotor response in varying CO2 concentrations, as evaluated using cine phase contrast MRI: Flow, volume, and cerebrovascular resistance indices

PURPOSE Previous studies have identified that impaired cerebral vasomotor reactivity (VMR) is associated with a higher risk of stroke and transient ischemic attack. This study aims to evaluate VMR by measuring the blood flow waveforms of the supplying arteries and dural sinuses using cine phase contrast MRI (PC MRI) and hypercapnic challenge. METHODS PC MRI flow quantification was performed on an oblique slice approximately perpendicular to the target vessels to include the left (LICA) and right internal carotid artery (RICA), basilar artery (BA), sinus rectus (SR), and superior sagittal sinus (SSS). A total of four PC MRI scans were performed at different CO(2) concentrations (room air and 3%, 5%, and 7% CO(2)). RESULTS The analyses obtained the flow parameters and cerebrovascular resistance parameters for all five vessels. Results indicated that the vascular resistance indices decreased with increasing CO(2) concentration in four vessels (LICA, RICA, BA, and SR). The obtained VMR parameters demonstrated exponential increases with increasing CO(2) concentration. CONCLUSIONS Using entire blood flow waveforms, this study applied separate flow dynamics during systolic and diastolic periods to obtain cerebrovascular resistance parameters and extensive flow-related information. It is the first to investigate the cerebrovascular resistance parameters under hypercapnic challenge using cine MRI. This technique could provide a useful tool for clinical application in cerebrovascular disease.

A Potential Risk of Overestimating Apparent Diffusion Coefficient in Parotid Glands

Objectives To investigate transient signal loss on diffusion weighted images (DWI) and overestimation of apparent diffusion coefficient (ADC) in parotid glands using single shot echoplanar DWI (EPDWI). Materials and Methods This study enrolled 6 healthy subjects and 7 patients receiving radiotherapy. All participants received dynamic EPDWI with a total of 8 repetitions. Imaging quality of DWI was evaluated. Probability of severe overestimation of ADC (soADC), defined by an ADC ratio more than 1.2, was calculated. Error on T2WI, DWI, and ADC was computed. Statistical analysis included paired Student t testing and Mann-Whitney U test. A P value less than 0.05 was considered statistically significant. Results Transient signal loss was visually detected on some excitations of DWI but not on T2WI or mean DWI. soADC occurred randomly among 8 excitations and 3 directions of diffusion encoding gradients. Probability of soADC was significantly higher in radiotherapy group (42.86%) than in healthy group (24.39%). The mean error percentage decreased as the number of excitations increased on all images, and, it was smallest on T2WI, followed by DWI and ADC in an increasing order. Conclusions Transient signal loss on DWI was successfully detected by dynamic EPDWI. The signal loss on DWI and overestimation of ADC could be partially remedied by increasing the number of excitations.

Laser direct writing 3D structures for microfluidic channels: flow meter and mixer

The 3D laser direct-writing technology is aimed at the modeling of arbitrary three-dimensional (3D) complex microstructures by scanning a laser-focusing point along predetermined trajectories. Through the perspective technique, the details of designed 3D structures can be properly fabricated in a microchannel. This study introduces a direct reading flow meter and a 3D passive mixer fabricated by laser direct writing for microfluidic applications. The flow meter consists of two rod-shaped springs, a pillar, an anchor, and a wedge-shaped indicator, installed inside a microfluidic channel. The indicator is deflected by the flowing fluid while restrained by the spring to establish an equilibrium indication according to the flow rate. The measurement is readily carried out by optical microscopy observation. The 3D passive Archimedes-screw-shaped mixer is designed to disturb the laminar flow 3D direction for enhancing the mixing efficiency. The simulation results indicate that the screw provides 3D disturbance of streamlines in the microchannel. The mixing demonstration for fluids flowing in the micrchannel approximately agrees with the simulation result. Thanks to the advantage of the laser direct writing technology, this study performs the ingenious applications of 3D structures for microchannels.

Recent advances in two-photon 3D laser lithography with self-Q- switched Nd:YAG microchip lasers

We review our recent results towards the development of a turnkey 3D laser printer, based on self-Q-switched microchip Nd:YAG lasers, with reproducible sub-100nm resolution, and with large-scale (cm) and fast-speed (cm/sec) capability at micron resolution. First of all, we report on line fabrication with 70nm lateral, and 150nm longitudinal resolutions without significant shrinking. This is due to the tight focusing with green visible wavelength, large numerical aperture, and excellent resin properties. Secondly, we report on two-photon sensitive photoacid generators that lead to efficient 3D microfabrication with epoxy SU-8 resin. Thirdly, we demonstrate high-speed microfabrication of large scale, millimeter size, scaffolds and cemtimeter height needle with high repetition rate (130Khz), and high average power (1W) amplified microchip laser. Finally we demonstrate the two-photon induced cross-linking of antibodies to determine the type of red blood cells in microfluidic channels.

Proton change of parotid glands after gustatory stimulation examined by magnetic resonance imaging

The aim of this study was to investigate proton changes of the parotid gland after gustatory stimulation by semi‐quantitative parameters and an empirical mathematical model (EMM) using high‐temporal‐resolution, double‐echo, echo‐planar imaging (EPI). Approved by a local institutional review board, this study examined 20 parotid glands from 10 healthy volunteers (male:female = 6: 4; age ± standard deviation =35.1 ± 14.1 years) with written informed consent obtained. All participants underwent 1.5‐T, double‐echo EPI with gustatory stimulation. Semi‐quantitative parameters, including maximal drop ratio (MDR), time to peak (TTP), drop slope (DS), recovery slope (RS) and recovery ratio (RR), were calculated. The effect of temporal resolution on parotid functional parameters was evaluated. An EMM comprising an output function ( Sot=Aoe−kot+B ) and an input function ( Sint=Ain1−e−kint ) was also applied to fit all dynamic curves. Kruskal–Wallis test, Wilcoxon test, linear regression analysis and goodness of fit were used for statistical analysis. p < 0.05 was considered to be statistically significant. The signal intensity dropped significantly after gustatory stimulation on the proton density (PD) image (p < 0.01). MDR was 8.26% in the PD image. MDR and RR were negatively associated with time interval, whereas DS and TTP were significantly positively associated with time interval (all p < 0.05). EMM parametric values derived from PD–time curves of parotid glands were 12.04 ± 6.81%, 6.43 ± 4.23 min–1, 88.73 ± 6.18%, 8.41 ± 4.86 min–1 and 1.09 ± 1.35 for Ao, ko, B, Ain and kin, respectively. Semi‐quantitative functional parameters and EMM parameters using high‐temporal‐resolution, double‐echo EPI allow the quantification of parotid proton changes after gustatory stimulation.

3D printing of natural organic materials by photochemistry

In previous works, we have used two-photon induced photochemistry to fabricate 3D microstructures based on proteins, anti-bodies, and enzymes for different types of bio-applications. Among them, we can cite collagen lines to guide the movement of living cells, peptide modified GFP biosensing pads to detect Gram positive bacteria, anti-body pads to determine the type of red blood cells, and trypsin columns in a microfluidic channel to obtain a real time biochemical micro-reactor. In this paper, we report for the first time on two-photon 3D microfabrication of DNA material. We also present our preliminary results on using a commercial 3D printer based on a video projector to polymerize slicing layers of gelatine-objects.