Biomedical Application of PHBV/Collagen (PHCP) Nanofiber as the Wound Dressing

Xu Zi Guang,문찬미,곽정식,오기완,배한익,한진이 충북대학교 동물의학연구소 2011 Journal of Biomedical and Translational Research Vol.12 No.3

In this study, a nanofibrous scaffold was obtained by co-electrospinning poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and collagen in 2,2,2-trifluoroethanol at a ratio of 3/7. The fiber diameters were in the range of 250-600 nm. It was found that PHBV/Collagen (PHCP) nanofibrous scaffold more proliferated than on the PHBV nanofibrous scaffold induced by oxidant in NIH3T3 cells. Otherwise, on the early-stage wound-healing mouse model, wound closure was evaluated by wound size reduction and histology of the regenerated skin on the back of mouse. An each of tissues removed on 0, 3, 6, 9, 12, 15 and 18 day were used to analyse biochemical and pathological changes. Every nanofiber-attached mice showed no significant difference to the 3th day, but from the third day until the nineth day, PHCP-attached mice were found to heal much faster than that of control wounds in epithelialisation, wound contraction and histopathological examinations. These results strongly support the beneficial effects of the biomedical application of PHCP nanofiber in the acceleration of wound healing initial phase through α-SM actin contraction.

Anti-inflammatory and Anti-oxidant Effects of PHBV/Collagen (PHCP) on Lipopolysaccharide-Induced Skin Inflammation

Xu Zi Guang,Jyung-Sik Kwak,Ki Wan Oh,배한익,한진이 충북대학교 동물의학연구소 2012 Journal of Biomedical and Translational Research Vol.13 No.1

The anti-inflammatory effect of PHBV/Collagen (PHCP) was examined in a mouse model of lipopolysaccharide (LPS)-induced skin inflammation. Vascular permeability on the back skin was measured by the local accumulation of evan’s blue dye after subcutaneous injection of LPS (30 μg site-1). Dye leakage in the skin was significantly increased 2 h after injection of LPS. This LPS-induced dye leakage was also completely inhibited by HO-1 inhibitor, ZnPP, and antioxidants, including methyl gallate, trolox, mannitol. To study the possible mechanisms underlying the in vivo anti-inflammatory effect of PHCP against LPS-induced inflammation, we also examined the effects of PHCP on malondialdehyde (MDA) and glutathione levels in skin tissues and found that PHCP pretreatment inhibited MDA elevation and the remarkable reduction of glutathione level. In addition, similar results were obtained after pretreatment with antioxidants such as trolox and mannitol and HO-1 inhibitor, ZnPP. Histopathologically, an influx of neutrophils into the skin dermis were detected between 24 h and 72 h after LPS injection (30, 100 μg site-1), compared to control animals after the injection of saline. This increase was greater in mice treated with 100 μg of mice than those of the 30 μg of mice and was significantly suppressed by pretreatment with PHCP, antioxidants and HO-1 inhibitor. These results collectively suggest that PHCP has anti-inflammatory effect against LPS-induced inflammation model in vivo and may be a good candidate for the skin tissue engineering biomedical application primarily through manipulation of the redox state.

Anti-inflammatory and Anti-oxidant Effects of PHBV/Collagen (PHCP) on Lipopolysaccharide-Induced Skin Inflammation

Jin-Yi Han,Xu Zi Guang,Jyung-Sik Kwak,Ki-Wan Oh,Han-Ik Bae 충북대학교 동물의학연구소 2012 Journal of Biomedical and Translational Research Vol.13 No.1

The anti-inflammatory effect of PHBV/Collagen (PHCP) was examined in a mouse model of lipopolysaccharide (LPS)-induced skin inflammation. Vascular permeability on the back skin was measured by the local accumulation of Evan’s blue dye after subcutaneous injection of LPS (30 µg site^-1 ). Dye leakage in the skin showed a significant increase at 2 h after injection of LPS. This LPS-induced dye leakage was also completely inhibited by HO-1 inhibitor, ZnPP, and antioxidants, including methyl gallate, trolox, and mannitol. To study the possible mechanisms underlying the in vivo anti-inflammatory effect of PHCP against LPS-induced inflammation, we also examined the effects of PHCP on malondialdehyde (MDA) and glutathione levels in skin tissues and found that pretreatment with PHCP resulted in inhibited MDA elevation and a remarkable reduction of glutathione level. In addition, similar results were obtained after pretreatment with antioxidants, including trolox and mannitol, and HO-1 inhibitor, ZnPP. Histopathologically, an influx of neutrophils into the skin dermis was detected between 24 h and 72 h after LPS injection (30, 100 µg site^-1), compared to control animals after injection of saline. This increase was greater in mice treated with 100 µg of LPS than in those treated with 30 µg of LPS and was significantly suppressed by pretreatment with PHCP, antioxidants, and HO-1 inhibitor. These results collectively suggest that PHCP has an anti-inflammatory effect against LPS-induced inflammation model in vivo and may be a good candidate for the skin tissue engineering biomedical application primarily through manipulation of the redox state.

Anti-inflammatory and Anti-oxidant Effects of PHBV/Collagen (PHCP) on Lipopolysaccharide-Induced Skin Inflammation

Jin-Yi Han3*, Xu Zi Guang, Jyung-Sik Kwak, Ki-Wan Oh, Han-Ik Bae 충북대학교 동물의학연구소 2012 Journal of Biomedical and Translational Research Vol.13 No.1

The anti-inflammatory effect of PHBV/Collagen (PHCP) was examined in a mouse model of lipopolysaccharide (LPS)-induced skin inflammation. Vascular permeability on the back skin was measured by the local accumulation of Evan’s blue dye after subcutaneous injection of LPS (30 μg site-1). Dye leakage in the skin showed a significant increase at 2 h after injection of LPS. This LPS-induced dye leakage was also completely inhibited by HO-1 inhibitor, ZnPP, and antioxidants, including methyl gallate, trolox, and mannitol. To study the possible mechanisms underlying the in vivo anti-inflammatory effect of PHCP against LPS-induced inflammation, we also examined the effects of PHCP on malondialdehyde (MDA) and glutathione levels in skin tissues and found that pretreatment with PHCP resulted in inhibited MDA elevation and a remarkable reduction of glutathione level. In addition, similar results were obtained after pretreatment with antioxidants, including trolox and mannitol, and HO-1 inhibitor, ZnPP. Histopathologically, an influx of neutrophils into the skin dermis was detected between 24 h and 72 h after LPS injection (30, 100 μg site-1), compared to control animals after injection of saline. This increase was greater in mice treated with 100 μg of LPS than in those treated with 30 μg of LPS and was significantly suppressed by pretreatment with PHCP, antioxidants, and HO-1 inhibitor. These results collectively suggest that PHCP has an anti-inflammatory effect against LPS-induced inflammation model in vivo and may be a good candidate for the skin tissue engineering biomedical application primarily through manipulation of the redox state.

Biomedical Application of PHBV/Collagen (PHCP) Nanofiber as the Wound Dressing

Jin-Yi Han,Xu Zi Guang,Chan-Mi Moon,Jyung-Sik Kwak,Ki-Wan Oh,Han-Ik Bae 충북대학교 동물의학연구소 2011 Journal of Biomedical and Translational Research Vol.12 No.3

In this study, a nanofibrous scaffold was obtained by co-electrospinning poly (3-hydroxybutyrate- co-3-hydroxyvalerate) (PHBV) and collagen in 2,2,2-trifluoroethanol at a ratio of 3/7. The fiber diameters were in the range of 250-600 nm. It was found that PHBV/Collagen (PHCP) nanofibrous scaffold showed greater proliferation than the PHBV nanofibrous scaffold induced by oxidant in NIH3T3 cells. Otherwise, in the early-stage wound-healing mouse model, wound closure was evaluated according to wound size reduction and histology of regenerated skin on the backs of mice. Each of the tissues removed on day 0, 3, 6, 9, 12, 15, and 18 was used for analysis of biochemical and pathological changes. None of the nanofiber-attached mice showed significant difference on the third day, however, from the third day until the ninth day, significantly faster healing was observed in PHCP-attached mice, compared to control wounds in epithelialization, wound contraction, and histopathological examinations. These results strongly support the beneficial effects of biomedical application of PHCP nanofiber in acceleration of the initial phase of wound healing through α-SM actin contraction.

Cloning and Molecular Characterization of a Novel Rolling-Circle Replicating Plasmid, pK1S-1, from Bacillus thuringiensis subsp. kurstaki K1

Ming Shun Li,Jong Yul Roh,Xueying Tao,Zi Niu Yu,Zi Duo Liu,Qin Liu,Hong Guang Xu,Hee Jin Shim,Yang-Su Kim,Yong Wang,Jae Young Choi,Yeon Ho Je 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.05

Bacillus thuringiensis, an entomopathogenic bacterium belonging to the B. cereus group, harbors numerous extra-chromosomal DNA molecules whose sizes range from 2 to 250 kb. In this study, we used a plasmid capture system (PCS) to clone three small plasmids from B. thuringiensis subsp. kurstaki K1 using PCS which were not found in B. thuringiensis subsp. kurstaki HD-1, and determined the complete nucleotide sequence of plasmid pK1S-1 (5.5 kb). Of the six putative open reading frames (ORF2-ORF7) in pK1S-1, ORF2 (MobK1) showed approximately 90% aa identity with the Mob-proteins of pGI2 and pTX14-2, which are rolling circle replicating group VII (RCR group VII) plasmids from B. thuringiensis. In addition, a putative origin of transfer (oriT) showed 95.8% identity with those of pGI2 and pTX14-2. ORF3 (RepK1) showed relatively low aa identity (17.8-25.2%) with the Rep protein coded by RCR plasmids, however. The putative double-strand origin of replication (dso) and single-strand origin of replication (sso) of pK1S-1 exhibited approximately 70% and 64% identities with those of pGI2 and pTX14-2. ORF6 and 7 showed greater than 50% similarities with alkaline serine protease, which belongs to the subtilase family. The other 2 ORFs were identified as hypothetical proteins. To determine the replicon of pK1S-1, seven subclones were contructed in the B. t huringiensis ori-negative pHT1K vector and were electroporated into a plasmid cured B. thuringiensis strain. The 1.6 kb region that included the putative ORF3 (Rep1K), dso and ORF4, exhibited replication ability. These findings identified pK1S-1 as a new RCR group VII plasmid, and determined its replication region.

Establishment and Analysis of Erosion Depth Model for Impeller Material FV520B

Zi-Wu Liu,Jian-Feng Li,Xiu-Jie Jia,Guang-Cun Wang,Wen-Han Xu 한국정밀공학회 2016 International Journal of Precision Engineering and Vol.3 No.1

The multifactorial erosion was conducted in this paper to test the compressor impeller material FV520B using high-speed gas-solid two phase flow erosion tester and surface morphology analysis method. Based on the particle motion and collision energy equation as well as regression analysis of multi-factor orthogonal experiment, a phenomenological erosion depth model which captures the effects of impact velocity, angle and particle size, has been developed. The model includes removal of material due to both deformation damage and micro-cutting. Results show that the peak of experiment depth and the maximum calculated depth all appeared at near 45o, rather than near 24o where the maximum erosion rate appeared. Comparing the calculated values and the results of each single factor experiment, the errors are within 15%. The predictions of the simplified version of the model were in good agreement with the results of single factor experiments. Also, the reliability of the assessment formula was verified to assess the impeller erosion life, which indicated that this calculation model could be used to estimate the erosion depth of compressor impeller material FV520B.

Effects of Collection Methods on Recovery Efficiency, Maturation Rate and Subsequent Embryonic Developmental Competence of Oocytes in Holstein Cow

Wang, Zheng-guang,Yu, Song-dong,Xu, Zi-rong Asian Australasian Association of Animal Productio 2007 Animal Bioscience Vol.20 No.4

Holstein cow ovaries obtained at a slaughterhouse were used to study the influence of the oocyte collection methods (slicing, puncture, aspiration I and II) on recovery efficiency and subsequent in vitro maturation and embryonic development competence of immature oocytes recovered. In the slicing method, the whole ovarian was chopped into small pieces with a surgical blade. In the puncture method, the whole ovarian surface was punctured by 18-g needle. In other 2 aspiration methods, collected oocytes by aspirating from the visible follicles using an 18-g needle attached to a 5 ml syringe (aspiration I) or using a constant negetive pressure (-80 mmHg) with a vacuum pump (aspiration II). The oocytes were classified into 4 classes on the basis of the morphology of cumulus cells and cytoplasmic appearance of oocyte. Slicing ($9.6{\pm}0.4$) and puncture ($9.7{\pm}0.4$)yielded a larger number of oocytes per ovary than other two aspiration methods (aspiration I and II were $5.8{\pm}0.3$and $5.6{\pm}0.4$, respectively) (p<0.05). The number of the highest quality oocytes (grade A) per ovary was significantly higher in slicing ($4.2{\pm}0.2$) and puncture ($4.6{\pm}0.1$) methods than in other methods (aspiration I and II were $1.2{\pm}0.2$ and $1.4{\pm}0.2$, respectively) (p<0.05). The rate of nuclear maturation of the highest and higher quality oocytes (grade A and grade B, respectively) was not affected by the oocytes collection methods. The oocytes collection methods also did not influence subsequent embryonic developmental competence after in vitro fertilization with M II stage oocytes. It is concluded that slicing and puncture methods of the ovaries can be used as an alternative techniques to aspiration by the syringe or vacuum pump.

Cloning and Molecular Characterization of a Novel Rolling-Circle Replicating Plasmid, pK1S-1, from Bacillus thuringiensis subsp. kurstaki K1

Ming Shun Li,노종열,Xueying Tao,Zi Niu Yu,Zi Duo Liu,Qin Liu,Hong Guang Xu,심희진,김양수,왕용,최재영,제연호 한국미생물학회 2009 The journal of microbiology Vol.47 No.4

Bacillus thuringiensis, an entomopathogenic bacterium belonging to the B. cereus group, harbors numerous extra-chromosomal DNA molecules whose sizes range from 2 to 250 kb. In this study, we used a plasmid capture system (PCS) to clone three small plasmids from B. thuringiensis subsp. kurstaki K1 which were not found in B. thuringiensis subsp. kurstaki HD-1, and determined the complete nucleotide sequence of plasmid pK1S-1 (5.5 kb). Of the six putative open reading frames (ORF2-ORF7) in pK1S-1, ORF2 (MobK1) showed approximately 90% aa identity with the Mob-proteins of pGI2 and pTX14-2, which are rolling circle replicating group VII (RCR group VII) plasmids from B. thuringiensis. In addition, a putative origin of transfer (oriT) showed 95.8% identity with those of pGI2 and pTX14-2. ORF3 (RepK1) showed relatively low aa identity (17.8~25.2%) with the Rep protein coded by RCR plasmids, however. The putative double- strand origin of replication (dso) and single-strand origin of replication (sso) of pK1S-1 exhibited approximately 70% and 64% identities with those of pGI2 and pTX14-2. ORF6 and 7 showed greater than 50% similarities with alkaline serine protease, which belongs to the subtilase family. The other 2 ORFs were identified as hypothetical proteins. To determine the replicon of pK1S-1, seven subclones were contructed in the B. thuringiensis ori-negative pHT1K vector and were electroporated into a plasmid cured B. thuringiensis strain. The 1.6 kb region that included the putative ORF3 (Rep1K), dso and ORF4, exhibited replication ability. These findings identified pK1S-1 as a new RCR group VII plasmid, and determined its replication region.

Prognostic Significance of Overexpression of EZH2 and H3k27me3 Proteins in Gastric Cancer

He, Long-Jun,Cai, Mu-Yan,Xu, Guo-Liang,Li, Jian-Jun,Weng, Zi-Jin,Xu, Da-Zhi,Luo, Guang-Yu,Zhu, Sen-Lin,Xie, Dan Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.7

The enhancer of zeste homolog 2 (EZH2) methyl transferase and histone 3 lysine 27 (H3K27me3) protein can repress gene transcription, and their aberrant expression has been observed in various human cancers. This study determined their expression levels in gastric cancer tissues with reference to clinicopathological features and patient survival. We collected 117 gastric cancer and corresponding normal tissues for immunohistochemistry analysis. In gastric cancers, 82/117 (70.1%) were positive for EZH2 and 66/117 (56.4%) for H3K27me3 proteins in contrast to only 5.41% and 7.25% of normal gastric mucosa specimens, respectively. Kaplan-Meier survival data showed the average overall and disease-free survival of EZH2 high expression patients was 25.2 and 20.2 months, respectively, shorter than that with EZH2 low expression (40.5 and 35.9 months). The average overall survival and disease-free survival of high H3K27me3 expression patients was 23.4 and 17.4 months, shorter than without H3K27me3 expression (37.6 and 34.5 months). The average overall survival and disease-free survival of patients with both EZH2 and H3K27me3 expression was 18.8 and 12.9 months, respectively, shorter than that with either alone (34.7 and 31.2 months) or with low levels of both (43.9 and 39.9 months). Multivariate Cox regression analysis showed that H3K27me3 and EZH2 expression, tumor size differentiation and clinical stage were all independent prognostic factors for predicting patient survival. This study demonstrated that detection of both EZH2 and H3K27me3 proteins can predict poor survival of gastric cancer patients, superior to single protein detection. In addition, H3K27me3 and EZH2 protein expression could predict lymph node metastasis.