Browsing by Author "Huang, MC"

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Effects of glucose concentration on in vitro fertilization in BALB/c mice
(2003-12-01)
Wu, HT
;
Chou, CK
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Lin, CS
;
Huang, MC
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生物科技學系
BALB/c mice are widely used in genetic, tumour and immunological studies. However, the mice demonstrate a lower reproduction rate, low fertility and small litters, because of their highly genetic homozygoisty. Based on in vitro fertilization (IVF), a routine technique for biomedical studies, it is worth to evaluate the effects to BALB/c mice on IVF efficiency. In order to test the genetic factor affecting the IVF efficiency of BALB/c, four reciprocal IVF tests of BALB/cByJ and FVB/NCrl mice were performed. The results showed that the average fertility of IVF sponsored by FVB/NCrl spermatozoa was 69.6%, but only 12.1% was obtained from BALB/cByJ strain. Effect of glucose contained in the culture medium to the IVF efficiency of BALB/cByJ was also evaluated. The results showed that the fertility of BALB/cByJ spermatozoa incubated with 0, 2.7, 5.5, 11.1 and 22.2 mM of glucose in the TYH medium were 6.8, 9.9, 13.9, 32.7 and 22.2%, respectively. It is showed that IVF efficiency of BALB/cByJ spermatozoa could be improved depending on the concentration of glucose in the IVF medium. According to the results, it is beleived that lower IVF of BALB/cByJ mice might be due to the genetic defect in spermatozoa and increasing glucose in the IVF medium which significantly affect the IVF efficiency of BALB/cByl via activating the spermatozoa.
Highly reliable chemical-mechanical polishing process for organic low-k methylsilsesquioxane
(2001-07-01)
Liu, PT
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Chang, TC
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Huang, MC
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Tsai, MS
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Sze, SM
In this work, chemical-mechanical polishing (CMP) of the organic polymer, methylsilsesquioxane (MSQ), has been investigated. For conventional silicate-based slurry, the CMP removal rate of MSQ is low and many scratches are formed at the surface. Moreover, the dielectric properties of a post-CMP MSQ film are degraded in comparison to the as-cured MSQ. We have proposed a reliable process for the CMP of MSQ which includes a slurry of additive and a post-CMP NH3 plasma treatment. Experimental results show that the modified slurry provides a high polishing rate and uniform surface topography. In addition, the NH3 plasma process can form a thin nitrogen-containing layer on the post-CMP MSQ surface, which enhances the resistance to moisture absorption and copper diffusion. (C) 2001 American Vacuum Society.
Improvement of post-chemical mechanical planarization characteristics on organic low k methylsilsesquioxane as intermetal dielectric
(2000-11-01)
Liu, PT
;
Chang, TC
;
Huang, MC
;
Yang, YL
;
Mor, YS
This work has investigated the electrical and material characteristics of post-chemical mechanical planarization (CMP) methylsilsesquioxane (MSQ). Experimental results have shown that the dielectric properties of low k MSQ deteriorate after the CMP process. However, by applying H-2-plasma post-treatment, the degraded characteristics can be restored to a similar state as that of a pre-CMP MSQ film. Material and electrical analyses were performed to elucidate the detailed mechanisms of H-2-plasma treatment on post-CMP MSQ. H-2-plasma treatment provides active hydro en radicals to passivate the dangling bonds exposed in the MSQ after the CMP process. The hydrogen-rich passivation layer is hydrophobic and effectively prevents further moisture uptake. Therefore, a degradation-free CMP process can be achieved employing H-2-plasma treatment. (C) 2000 The Electrochemical Society. S0013-4651(00)04-098-2. All rights reserved.
In vitro and ex vivo green fluorescent protein expression in alveolar mammary epithelial cells and mammary glands driven by the distal 5 '-regulative sequence and intron 1 of the goat beta-casein gene
(2003)
Wu, HT
;
Lin, CS
;
Huang, MC
;
生物科技學系
;
Department of Biological Science and Technology
The 5'-regulative sequence and intron 1 of the goat beta-casein gene from -4044 to +2123 by was cloned and fused with the reporter gene of green fluorescent protein (GFP) to create a plasmid termed pGB562/GFP To detect GFP expression, pGB562/GFP was transfected in vitro via liposomes into the mammary epithelial cell line NMuMG. Cells could not express GFP unless the transfected NMuMG cells lined up to create functional alveoli. These functional cells were cultured with lactogenic hormones, including insulin, dexamethasone and prolactin, and were grown on a layer of the extracellular matrix Matrigel. Green fluorescent protein expression levels in NMuMG cells were 25-, 55- and 42-fold those in the control group at 24, 48, and 72 h after pGB562/GFP transfection respectively. In addition, pGB562/GFP was transfected ex vivo by electroporation into mammary gland fragments and cells were then cultured in vitro with a supplement of lactogenic hormones. Strong GFP expression localized in fragments of the mammary gland was observed 24 h after gene transfer. The novel strategy of ex vivo gene transfer into mammary tissue using GFP as a reporter gene to detect the function of a tissue-specific promoter is efficient and convenient. The data obtained herein reveal that the 5'-regulative sequence and intron 1 of the 6.2 kb goat beta-casein gene can enhance the efficiency of transgene expression. Thus, the GB562 sequence may act as a good promoter and effectively elevate the production of exogenous protein in mammary glands.
Postnatal expression of growth/differentiation factor-8 (GDF-8) gene in European and Asian pigs
(2002-09-01)
Lin, CS
;
Wu, YC
;
Sun, YL
;
Huang, MC
;
生物科技學系
Myostatin (growth differentiation factor (GDF)-8), is one member of the transforming growth factor beta superfamily. Investigations of GDF-8 null mice and double-muscled cattle revealed that GDF-8 has a profound influence upon skeletal muscle growth. Therefore, the GDF-8 effect upon the productive performance of pigs is worth exploring. In the present study, the nucleotide sequences and expression levels of GDF-8 genes in European pigs (Landrace and Duroc) and Asian pigs (Taoyuan and Small-ear) were evaluated. Based upon their genetic background these breeds possess significantly distinct growth rate and muscle production-phenotypes. Our sequence data showed that the nucleotide sequences of European and Asian pigs were 100% similar. Postnatal expression of GDF-8 gene in skeletal muscles, from birth to 12 mo of age, among different breeds was measured. GDF-8 expression levels in the longissimus muscle of neonatal European breed littermates were the highest, however it declined significantly (p<0.05) at 1 and 3 mo, and then increased gradually at 6 to 12 mo. The Asian breeds, however, GDF-8 expression level increased markedly at 3 mo and maintained a constant level thereafter. The results indicate that rather than polymorphism within the GDF-8 functional sequence between European and Asia breeds, it was relative to the gene regulation in postnatal muscle growth.