Research Express@NCKU - Articles Digest
Research Express@NCKU Volume 19 Issue 5 - August 12, 2011
[ http://research.ncku.edu.tw/re/articles/e/20110812/1.html ]
Intramyocardial peptide nanofiber injection improves
post-infarction ventricular remodeling and efficacy of
bone marrow cell therapy in pigs
Patrick C.H. Hsieh
Assistant Professor & Attending Surgeon
Graduate Institute of Clinical Medicine and Center for Clinical Research, Department of Surgery &
Graduate Institute of Biomedical Engineering, National Cheng Kung University & Hospital, Tainan,
Taiwan
[email protected]
A
bstract
Background• Growing evidence suggests that intramyocardial biomaterial injection
improves cardiac functions after myocardial infarction (MI) in rodents. Cell therapy is
another promising approach to treat MI, although poor retention of transplanted cells is a
major challenge. In this study, we hypothesized that intramyocardial injection of selfassembling peptide nanofibers (NFs) thickens the infarcted myocardium and increases
transplanted autologous bone marrow mononuclear cell (MNC) retention to attenuate cardiac remodeling and
dysfunction in a pig MI model.
Methods and Results• A total of 40 mature minipigs were divided into 5 groups: sham, MI+normal saline (NS), MI
+NFs, MI+MNCs, and MI+MNCs/NFs. MI was induced by coronary occlusion, followed by intramyocardial
injection of 2 ml NS or 1% NFs with or without 1×108 isolated autologous MNCs. NF injection significantly
improved diastolic function and reduced ventricular remodeling 28 days after treatment. Injection of MNCs alone
ameliorated systolic function only, while injection of MNCs with NFs significantly improved both systolic and
diastolic functions, as indicated by +dP/dt and –dP/dt (1214.5±91.9 and -1109.7±91.2 mmHg/s in MI+NS, 1693.7
±84.7 and -1809.6±264.3 mmHg/s in MI+MNCs/NFs, respectively), increased transplanted cell retention (29.3
±4.5 cells/mm2 in MI+MNCs and 229.4±41.4 cells/mm2 in MI+MNCs/NFs), and promoted capillary density in the
peri-infarct area.
Conclusions• We demonstrated that NF injection alone prevents ventricular remodeling, while cell implantation
with NFs improves cell retention and cardiac functions after MI in pigs. This unprecedented combined treatment in
a large animal model has therapeutic effects, which can be translated to clinical applications in the foreseeable
future.
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Figure 1. Injection of NFs increases IVS thickness, and injection of MNCs improves systolic functions after
infarction. A, Histogram of left ventricle ejection fraction (LVEF) immediately after and 28 days after MI in sham
and experimental groups. B, Statistical analysis of IVS thickness at systole and diastole. *vs. MI+NS; †vs. MI
+NFs; ‡vs. MI+MNCs. IVS, interventricular septum.
Figure 2. Injection of NFs prevents infarct expansion, and injection of MNCs improves tissue viability after
infarction. A, Representative cross-section images at the level of papillary muscle insertion of left ventricle from
each group. B, Percentages of scar thickness ratio, scar length ratio, and non-viable area ratio determined using
computer image analysis. *vs. MI+NS; †vs. MI+NFs; ‡vs. MI+MNCs.
Figure 3. Injection of NFs along with MNCs improves transplanted cell retention after MI. A, Representative
pictures of DiI+ cells (red) in the area injected with cells alone (left panel) or with cells and NFs (right panel).
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Nuclei were stained by DAPI (blue). B, Cell retention rates as reflected by DiI+ cell counts.
Figure 4. Injection of NFs along with MNCs improves vascular cells differentiation after infarction. A,
Representative images of red DiI fluorescence, overlapped with green immunostaining for von Willebrand factor
(upper panel), SM myosin heavy chain (middle panel), and cardiac tropomyosin (lower panel). Nuclei were stained
with DAPI (blue). The image (left panel) was magnified from right panel indicated in the boxes, and co-stained
cells were arrowed. B, Quantification of ratio of cells stained with endothelial cell (EC), smooth muscle cell
(SMC), and cardiomyocyte (CM) markers.
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Figure 5. Injection of NFs along with MNCs improves capillary density in the peri-infarct area. A, Representative
immunostaining of isolectin (green), overlapped with cardiomyocytes stained with cardiac tropomyosin (red) at the
border zone from each group. Nuclei were stained by DAPI (blue). B, Quantification of capillary density in the
peri-infarct zone. *vs. MI+NS; †vs. MI+NFs.
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Research Express@NCKU Volume 19 Issue 5 - August 12, 2011
[ http://research.ncku.edu.tw/re/articles/e/20110812/2.html ]
Altered Inflammatory Responses in Periventricular
Leukomalacia Preterm Children
Chang-Yi Lin1, Ying-Chao Chang2, Shan-Tair Wang3, Ting-Yang Lee4, Chiou-Feng Lin5 and
Chao-Ching Huang5,6,*
1
Institute of Basic Medical Sciences, National Cheng Kung University College of Medicine
Institute of Public Health,
4 Department of Biochemistry and Molecular Biology,
5 Institute of Clinical Medicine,
6 Department of Pediatrics,
2 Department of Pediatrics, Chang Gung Memorial Hospital-Kaohsiung Medical Center, Chang Gung
University College of Medicine, Kaohsiung
3
[email protected]
Ann. Neurol. 68, 204-212 Aug (2010)
I
ntroduction
Early life events play a powerful role in influencing later susceptibility to certain chronic
diseases. Injury occurring during critical periods of development, such as perinatal period,
may have a long-term effect on physiological function, which includes growth,
metabolism and inflammatory responses. Periventricular leukomalacia (PVL) is the major
form of brain injury and also the leading cause of spastic CP in preterm infants. Previous
studies have demonstrated that perinatal inflammatory responses contribute to PVL in preterm infants. In this
study, we used a cohort of 32 preterm children with PVL-induced CP (CP group) and their gestational age matched
32 preterm children with normal neurodevelopment (normal group) to test the hypothesis that preterm children
who develop PVL-induced spastic CP have an altered proinflammatory status due to increased lipopolysaccharide
(LPS) sensitivity in the peripheral blood mononuclear cells (PBMCs).
Results
There was also no significant difference in the mean ages of the 2 groups at the follow-up examination. The CP
and normal group were similar in birth-weight, and in the percentage of maternal toxemia, maternal hypertension,
use of prenatal dexamethasone, clinical chorioamnionitis, respiratory distress syndrome, surfactant use, patent
ductus arteriosus, bacteria/sepsis, and retinopathy of prematurity. Compared with the normal group, the CP group
had significantly higher frequency of chronic lung disease, and significantly lower in full IQ, verbal IQ, and
performance IQ scores. Due to suffering from PVL, PVL-induced CP group had significantly lower gross motor
scores than those of the control group.
We collected the plasma and PBMCs from these 2 groups. We first measured the plasma levels of TNF-α by
utilizing enzyme-linked immunosorbent assay kits. We found that plasma levels of TNF-•αwere significantly
higher in CP group children than in control group children (means±SEM: CP, 53±16pg/ml vs control, 10±4pg/ml;
p <0.001). Then, we utilized a LPS-stimulated PBMCs model to examine the extracellular and intracellular levels
of TNF-α •of PBMCs under LPS stimulation. The results showed that the CP group had significantly higher levels
of secreted TNF-α from LPS-stimulated PBMCs than those from the control group (CP: 1,736 ± 252pg/ml vs
control:1,031 ± 135pg/ml; p = 0.003). Apart from high levels of secreted TNF-α, flow cytometry analysis showed
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that after LPS stimulation, the levels of intracellular TNF-αwere significantly decreased in the control group (n =
13; geometric mean [Gm] before LPS, 1.9 ± 0.1; after LPS, 1.7 ± 0.1; p =0.016); in contrast, the levels were
significantly elevated in the CP group (n = 15; Gm before LPS, 1.6 ± 0.1; after LPS, 2.1 ± 0.2; p = 0.01)
We then examined whether the increased response of TNF-α in the LPS-stimulated PBMCs of the CP children was
related to the increased expression of inflammatory signaling molecules in their PBMCs. TNF superfamily genes
array (Superarray) and real-time PCR demonstrated that TLR-4, TAK1, JNK, IKK-r, and TNF-α mRNA
expression in the nonstimulated PBMCs were significantly higher in the CP group (n = 17) than those in the
control group (n = 13). In addition, the TLR-4 mRNA levels of the nonstimulated PBMCs were highly correlated
with the secreted protein levels of TNF-•α from the LPS-stimulated PBMCs
Conclusions and discussion
We provide the first evidence that plasma levels of TNF-α, levels of TNF-•α released from LPS-stimulated
PBMCs, and mRNA levels of inflammatory signaling molecules (TLR-4, TAK1, JNK, IKK-r, and TNF-α) in
PBMCs are significantly higher in school-age preterm children with PVL-induced CP than in normal control
school-age preterm children. In addition, after LPS stimulation, the intracellular TNF-•α level of the PBMCs was
significantly higher in the CP group, but significantly lower in the control group. This finding suggests that
preterm children with a remote PVL injury-induced CP have altered inflammatory responses.
Acknowledgements
This study was supported by grants from the Taiwan National Health Research Institute (NHRI-EX 95,96,979414NI), the National Science Counsel (NSC 97-2811-B-006-014, NSC98-2628-B006-001-MY3), and the Center
for Gene Regulation and Signal Transduction Research, National Cheng Kung University.
References
1. Yeh TF, Lin YJ, Lin HC, et al. Outcomes at school age after postnatal dexamethasone therapy for lung
disease of prematurity. N Engl J Med 2004;350:1304–1313.
2. Gluckman PD, Hanson MA, Cooper C, Thornburg KL. Effect of in utero and early-life conditions on adult
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health and disease. N Engl J Med 2008;359:61–73.
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Research Express@NCKU Volume 19 Issue 5 - August 12, 2011
[ http://research.ncku.edu.tw/re/articles/e/20110812/3.html ]
Binary liquid crystal alignments based on
photoalignment in azo dye-doped liquid crystals and
their application
Andy Ying-Guey Fuh1,2,3,*, Ju-Chin Chen1, Ko-Ting Cheng1 and San-Yi Huang2
1
Department of Physics, National Cheng Kung University
Institute of Electro-optical Science and Engineering, National Cheng Kung University
Advanced Optoelectronic Technology Center, National Cheng Kung University
2
3
[email protected]
Applied Physics Letters 96, 051103-1-3 (2010)
L
iquid crystal (LC) alignment on substrates is one of the key steps in the manufacturing
of LC devices. Commonly used LC alignments, including homogeneous alignment (HA),
homeotropic (vertical) alignment (VA), twisted nematics (TN), hybrid alignment (HB),
HA with pretilt angles, and others, are adopted to fabricate LC devices. However, many
LC devices require two or more forms of LC alignment to deliver the required optical
properties. Such devices include transflective LCDs, [1] LC holographic gratings, [2] and
others. In this paper, the term of “binary LC alignments” is defined as two different
common LC alignments in a specific area. Briefly, binary LC alignments, including orthogonally HA, orthogonally
HB, orthogonally TN, HA/VA, HA/HB, HA/TN, HB/TN, VA/HB and VA/TN alignments, can be adopted to
produce phase-modulated structures. [1-2] Photoalignment based on photoisomerization by the light-induced
adsorption of azo dyes, such as methyl red (MR) doped in LCs, has been widely discussed. [3-5] Additionally, the
authors previously examined that the adsorption rate of MR dyes onto a polymer surface, NOA81, markedly
exceeds that onto indium-tin-oxide (ITO)-coated glass substrates. [4] In this work, an approach for producing
binary LC alignments in azo dye-doped liquid crystals (ADDLCs) and their application to fabricate a viewingangle-dependent LCD are demonstrated.
The LC and the azo dye that were adopted herein were E7 (from Merck, 99wt%) and methyl red (MR, from
Aldrich, 1 wt%). A layer of prepolymer (NOA81) in acetone was spin-coated onto an ITO-coated glass substrate
that was covered with a homeotropic alignment film of DMOAP. After coating, the substrate was irradiated under
unpolarized UV light through a stripe-type photo-mask with alternating opaque and transparent stripes and a
spacing between stripes of the same kind of ~1.4 mm. Then, the prepolymers in the un-polymerized regions were
removed by immersing the substrate into a suitable solvent. UV-cured stripe-type polymer patterns were finally
generated on the DMOAP-coated ITO glass substrate. Each empty cell (cell gap ~8 μm) was fabricated by
combining one substrate (SC) with stripe-type polymer patterns and one ITO-coated glass substrate (SR) that was
covered with a homeotropic alignment film of DMOAP. The mixed material of ADDLCs was then injected into
the empty cell.
An ADDLC sample with stripe-type polymer patterns before and after being irradiated with linearly polarized
green laser light (EG, from DPSS laser, λG=532 nm) was observed under a cross-polarizers polarized optical
microscope (POM) before view-angle-dependent LCDs was demonstrated. Figures 1(a) [1(d)], 1(b) [1(e)] and 1(c)
[1(f)] present the POM images of the sample with the stripes at 0˚, 45˚ and 90˚, respectively, from the
transmission axis of the polarizer before (after) the ADDLC sample was illuminated by EG having an intensity of
50 mW/cm2 from SC for about 10 minutes. Notably, the excited azo dyes are only anisotropically adsorbed onto
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the striped regions with polymer, but not onto the DMOAP surface because of the difference between the MRadsorption rates onto NOA81 and DMOAP surfaces. [4] Therefore, HB and VA LC alignments are achieved in the
regions with and without polymers, respectively. Figure 2 displays the steps of the fabrication of the specially
designed picture, formed by combing by two pictures (Figs. 2(b) and 2(d)). Firstly, the picture in Fig. 2(b) was
striped (spacing is ~1.4 mm) with transparent lines to become that in Fig. 2(c). Then, the striped picture (Fig. 2(c))
was superimposed on another picture (Fig. 2(d)) to yield the specially designed picture (Fig. 2(a)). Finally, a
viewing-angle-dependent LCD was fabricated by placing the ADDLC sample with binary LC alignments of VA
and HB, sandwiched between two crossed polarizers with transmission axes that made ± 45˚ with the long axis of
the adsorbed MR, on the top of the specially designed picture. Notably, the HB alignment striped regions should
precisely match the stripes in Fig. 2(c).
Fig. 1. Images of fabricated ADDLC sample with stripe-type polymer patterns under cross-polarizers POM. (a)
[(d)], (b) [(e)], and (c) [(f)] are images of sample rotated through 0˚, 45˚ and 90˚, respectively, from the
transmission axis of the polarizer before (after) irradiation under linearly polarized green light. P and A represent
the transmission axes of the polarizer and analyzer, respectively. EG presents the direction of polarization of the
impinging beam.
Fig. 2. (a) Specially designed picture obtained by striping image (b) with transparent lines (separated by spacing
~1.4 mm) to yield (c). Combining (c) and (d) yields the specially designed picture (a).
Figure 3 shows the results observed in various viewing directions, (θ, ф), where θ and ф•are the polar angle and
the azimuthal angle, respectively. Figure 3(a) presents the image seen in the normal direction, θ=0˚. Clearly, the
LC alignment in the dark stripes in Fig. 3(a) is VA, while that in the bright stripes is HB, since phase retardation in
VA regions (dark) was absent (∆n=0), but in HB regions (bright) was presented (∆n≠0) at normal incidence under
±45˚ crossed polarizers. Therefore, the LCD displays the image in Fig. 2(d). Figure 3(b) shows the image
observed from a viewing direction of about θ=20˚ and ф=60˚. In such a case, the LC alignment in the bright
stripes is VA, while that in the dark stripes is HB. The cause of the light leakage in VA regions is the phase
retardation, 2πd(neff-no)/λ≠2nπ, and the cause of the darkness in HB regions is the phase retardation, 2πd(neff-no)/
λ=2nπ, where d, neff, λ and n are the cell gap, the effective refraction index of LCs, the wavelength of the incident
light, and an integer, respectively. Therefore, the LCD displays the image in Fig. 2(b). In a viewing direction of
about θ=30˚ and ф=180˚, the phase retardation in VA and HB regions are not 2nπ. Figures 3(a) and 3(b) can
be displayed simultaneously, as presented in Fig. 3(c). Additionally, when the viewing angle is around θ=85˚ and
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ф=225˚, all regions in the ADDLC sample are dark, as shown in Fig. 3(d). The results can be understood by the
fact that the phase retardations in these two LC alignments are 2nπ.
Fig. 3. Fabricated LCD observed in various viewing directions of (a) θ=0˚, (b) θ=20˚ and ф=60˚, (c) θ=30˚
and ф=180˚, and (d) θ=85˚ and ф=225˚, where θ and ф•are the polar angle and the azimuthal angle,
respectively.
The transmittance of incident light through an ADDLC sample that is sandwiched between two crossed polarizers
was also simulated using 1D-DIMOS software. Figures 4(a) and 4(b) present the simulated transmittance of VA
and HB ADDLC samples, respectively. The transmittances in the viewing directions of A(θ=0˚), B(θ=20˚,
ф=60˚), C(θ=30˚, ф=180˚) and D(θ=85˚, ф=225˚), marked in Figs. 4(a) and 4(b), are consistent with
those presented in Figs. 3(a)-3(d), respectively. Accordingly, a binary LC alignment (VA and HB) can be adopted
to fabricate a viewing-angle-dependent LCD. Notably, the visibility of the displays in Fig. 3 is poor. It can be
improved by using a photo-mask with a narrower spacing between stripes.
Fig. 4. Simulated transmittances of incident beam through (a) VA and (b) HB ADDLC samples at various viewing
angles. Polar angle (θ) and azimuthal angle (ф) in regions A, B, C and D are (θ=0˚), (θ=20˚ and ф=60˚),
(θ=30˚ and ф=180˚), and (θ=85˚ and ф=225˚), respectively, corresponding to Figs. 4(a), 4(b), 4(c) and 4
(d).
In conclusion, this work elucidates an approach for achieving various binary LC alignments using a surface-treated
alignment layer and a photoalignment film of the adsorption of azo dyes onto the polymer surface in ADDLCs.
The application of binary LC alignments (VA/HB) to fabricate a viewing-angle-dependent LCD was demonstrated,
and the experimental results match theoretical predictions.
Reference
[1] H.-Y. Kim, Z.-B. Ge, S.-T. Wu and S.-H. Lee, Appl. Phys. Lett. 91, 231108 (2007).
[2] C.-J. Yu, J.-H. Park, J. Kim, M.-S. Jung and S.-D. Lee, Mater. Sci. Eng. C 24, 247 (2004).
[3] F. Simoni, O. Francescangeli, Y. Reznikov and S. Slussarenko, Opt. Lett. 22, 549 (1997).
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[4] S.-Y. Huang, S.-T. Wu and Andy Y.-G. Fuh, Appl. Phys. Lett. 88, 041104 (2006).
[5] Andy Y.-G. Fuh, C.-C. Chen, C.-K. Liu and K.-T. Cheng, Opt. Express 17, 7088 (2009).
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Research Express@NCKU Volume 19 Issue 5 - August 12, 2011
[ http://research.ncku.edu.tw/re/articles/e/20110812/4.html ]
“Wild Boy” and “Perfect Lad”: The Construction of
Boyhood in Lu Pin Hua and A Huei De Xin
Mei-Ying Wu
Associate Professor of Department of Taiwanese Literature, College of Liberal Arts, National Cheng
Kung University
[email protected]
NTU Studies in Taiwan Literature 8(2010): 125-152
C
hung Chao-Cheng’s Lu Pin Hua (Lupin Flower) and Lin Chung-Lung’s A Huei De
Xin (A-huei’s Heart) are two of the initial publications of juvenile fiction in Taiwan. Lu
Pin Hua was serialized in The United Daily News in 1960 and later published by MingZhi Publisher in 1962, while A Huei De Xin was first serialized in Xiao Xue Sheng Za Zhi
(The Elementary Student Magazine) in 1964 and published a year later. Both are enlisted
in Taiwan Children’s Literature 100 (1945-1998), and accordingly each can be regarded
as one of the canonical children’s texts for young readers. Drawing on R. W. Connell’s
theory of hegemonic masculinity, as well as employing Western canonical boy stories for reference, the paper aims
to explore how boyhood and, in particular, how the ideal boy images and boy subjects are shaped and represented
in Taiwan juvenile fiction in the 1960s, specifically, in Lu Pin Hua and A Huei De Xin.
According to R. W. Connell, hegemonic masculinity can be defined as “the configuration of gender practice which
embodies the currently accepted answer to the problem of the legitimacy of patriarchy, which guarantees (or is
taken to guarantee) the dominant position of men and the subordination of women” (Masculinities 77). Following
Antonio Gramsci’s concept of hegemony in analyzing class relations, Connell focuses on the dynamic nature of
such a concept and emphasizes that hegemonic masculinity, in some cases, as the most valued and desired mode of
gender expression is not necessarily the most common one. Albeit hegemonic masculinity is often referred to male
domination or aggression in subordinating females or feminized others, it is important to understand that the term
hegemony is not to be taken as a top-down totalitarian coercion, but a subtle and saturated way to form common
sense and reach (sociopolitical) consensus. Take the discourse of boyhood for instance. “Boys will be boys” and
“boys don’t cry” are two of the common notions referring to boys’ gender significations. Such gender
significations for boys clearly illustrate what a boy should (not) be and what he should (not) do. However, a
discourse as such is problematic in forming and consolidating an idealized and normalized boy image that often
shapes and regulates our understanding and verification of boyhood.
A-Huei, the protagonist in A Huei De Xin, is a boy of
tremendous perseverance, self-control, and selfdiscipline. Gu A-Ming, the boy protagonist in Lu Pin
Hua, on the other hand, is the one characterized by
waywardness, self-assertion, and unconformity. Although
shaped in different ways—as A-Huei looks more like a
mature and sophisticated “grown-up,” whereas A-Ming
appears to be and behave like a “wild boy,” they
respectively represent the cultural expectations and
sanctions of what an ideal boy could be and should be.
The subordinating types of boys, such as Lin Zhi-Hong,
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Shui-Niu, and Su Hong-Fan, however, are foil characters
that function primarily to contest and negotiate with the
boy protagonists in terms of the display of hegemonic
masculinity. With the differentiated ideal boy images represented by the two boy protagonists, as well as the group
of subordinating boys, the construction of boyhood in the two early literary texts produced for young people in
Taiwan may point to a complex understanding of boy culture.
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