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Biocompatibility studies of silk fibroin-based artificial nerve grafts in vitro and in vivo

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Progress in Natural Science:Materials International 2007年第9期17卷 1029-1034页

作者： Chen Xuemei,Yang Yumin,Wu Jian,Zhao Yahong,Ding Fei and Gu Xiaosong(Jiangsu Key Laboratory of Neuroregeneration,nantong University,nantong 226001,china) Jiangsu Key Laboratory of Neuroregeneration Nantong University Nantong 226001 China

Silk fibroin(SF)has been used extensively in the biomedical field including tissue engineering for the generation of artificial bones,skins or *** have previously reported on good in vitro biocompatibility of SF fibers with peripheral nerve tissues and *** the present study,we developed a novel design of the SF-based artificial nerve graft(SF graft)which was composed of a SF-nerve guidance conduit(NGC)inserted with SF *** assay was performed to determine the cytotoxicity of the SF-NGC extract fluid on the cultured L929 cells derived from an immortalized mouse fibroblast cell *** addition,this SF graft was implanted into adult rats for bridging a 10-mm long sciatic nerve *** following-up experiments at initial stage(1-4 week)of nerve regeneration including routine blood tests and histochemical investigation were conducted to evaluate the in vivo biocompatibility of the SF graft with peripheral *** results demonstrated that the SF-NGC graft was biocompatible with the surrounding tissues and cells due to its low inflammatory potential with a grade 0 under the *** guidelines and it was generally suitable to a certain degree for bridging peripheral nerve defects in virtue of supporting Schwann cell adherence,expansion and *** the SF graft is a promising alternative to classical autografts for peripheral nerve repair.

关键词： silk fibroin artificial nerve graft nerve guidance conduit biocompatibility cytotoxicity

Application of Luxol Fast Blue staining in locating the corticospinal tract in adult rats

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Neural Regeneration Research 2006年第7期1卷 642-644页

作者： Su Liu1, Guangyu Shen1, Guangming Lü2, Xiaosong Gu3 1Department of Rehabilitation Medicine, Affiliated Hospital of nantong University, nantong 226001, Jiangsu Province, china 2Department of Human Anatomy, nantong University, nantong 226001, Jiangsu Province, china 3Jiangsu Key Laboratory of Nerve Regeneration, nantong 226001, Jiangsu Province, china Department of Rehabilitation Medicine Hospital of Nantong University Nantong 226001 Jiangsu Province China Department of Human Anatomy Nantong University Nantong 226001 Jiangsu Province China Jiangsu Key Laboratory of Nerve Regeneration Nantong 226001 Jiangsu Province China

BACKGROUND: There are many methods for myelin staining, mordant, or oil-soluble dye or the special reaction of osmic acid with lipoid is used according to different principles. The commonly used methods are classic Well staining, classic lithium carbonate-haematine staining, fast green staining, silver staining, etc. Luxol Fast Blue can brightly stain myelin sheath, and has certain specificity. The background can be very clean if there is proper differentiation, whereas Luxol Fast Blue is cheap and convenient to operate, thus it is an ideal staining reagent for routine myelin sheath. OBJECTIVE: To show the corticospinal tract of normal adult rats with Luxol Fast Blue staining method. DESIGN: A repetitive measurement design. SETTINGS: Institute of Nuerobiology, nantong University; Department of Rehabilitation Medicine, Affiliated Hospital of nantong University. MATERIALS: Six healthy adult male SD rats of clean degree, weighing averagely 300 g, were provided by the experimental animal center of nantong University. 1 g/L Luxol Fast Blue solution was provided by Sigma Company; Leica CM1900 cryostat microtome by Leica Company; Leica DMR microscope by Leica Company. METHODS: The experiment was carried out in the Staff Room of Human Anatomy, nantong University in May 2005. The rats were given intraperitoneal injection of combined anesthetic (2 mL/kg), then the chest was open for perfusing saline and phosphate buffer containing formamint via heart. Brain and spinal cord were removed after 1 hour then fixed, then changed to phosphate buffer (pH 7.4) containing 300 g/L saccharu at 4 ℃, and stayed overnight, tissue blocks at pyramid, decussation of pyramid and cervical, thoracic, lumbar and sacral segments of spinal cord were removed to prepare continuous horizontal frozen sections (30 μm) after sedimentation, the sections were dried at room temperature. The corticospinal tract of normal adult rats were shown with Luxol Fast Blue staining method, and observed under Leica DMR

关键词： Application of Luxol Fast Blue staining in locating the corticospinal tract in adult rats

Achyranthes bidentata Blume extract promotes neuronal growth in cultured embryonic rat hippocampal neurons

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Progress in Natural Science:Materials International 2009年第5期19卷 549-555页

作者： Xin Tang1, Yiren Chen 1, Xiaosong Gu, Fei Ding Jiangsu Key Laboratory of Neuroregeneration, nantong University, nantong 226001, china [a]Jiangsu Key Laboratory of Neuroregeneration Nantong University Nantong 226001 China

We have prepared an aqueous extract of Achyranthes bidentata Blume, a commonly prescribed Chinese medicinal herb, and reported, in previous studies, that A. bidentata extract benefits nerve growth and prevents neuron apoptosis. In this study, we investigated the actions of A. bidentata extract on survival and growth of primarily cultured rat hippocampal neurons. The morphological observation revealed that neurite growth from hippocampal neurons was significantly enhanced by A. bidentata extract with similar effects to those induced by nerve growth factor (NGF), and the greatest neurite growth appeared on treatment with A. bidentata extract at 1 lg/ml for 24 h. DNA microarray analysis indicated that there were 25 upregulated genes and 47 downregulated genes exhibiting significantly differential expression in hippocampal neurons treated with A. bidentata extract at 1 lg/ml for 6 h when compared to those in untreated hippocampal neurons. Real-time quantitative RT-PCR and Western blot analysis demonstrated that the expression of growth-associated protein-43 in hippocampal neurons was upregulated at both mRNA and protein levels after treatment with A. bidentata extract, and the optimal dosage of the extract was also 1 lg/ml. These data confirm that A. bidentata extract could promote in vitro hippocampal neuronal growth in a dose-and time-dependent manner.

关键词： Achyranthes bidentata Blume Aqueous extract Nerve growth factor Hippocampal neurons Neurite outgrowth Growth-associated protein-43 Gene differential expression

Abnormal expression of hepatoma- derived γ-glutamyltransferase subtyping and its early alteration for carcinogenesis of hepatocytes

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Hepatobiliary & Pancreatic Diseases International 2004年第4期3卷 564-570页

作者： Deng-Fu Yao, Zhi-Zhen Dong, Deng-Bing Yao, Xin-Hua Wu, Wei Wu, Li-Wei Qiu, Hong-Mei Wang and Xian-Yong Meng nantong, china Research Center of Clinical Molecular Biology, Affilia- ted Hospital of nantong University, nantong 226001 , china Insti- tute of Neuroscicnces, nantong University, nantong 226001 , china Research Center of Clinical Molecular Biology Affiliated Hospital of Nantong University Nantong 226001 China. dfyrcmb@***

BACKGROUND: Although the hepatoma-specific band of gamma-glutamyltransferase ( GGT ) is a highly sensitive marker in diagnosis of hepatocellular carcinoma, the kine- tic expression and the early alterations of GGT in the deve- lopment of hepatoma remain unclear. In this study, we in- vestigated the expression and the alterations of GGT multi- ple molecular forms in hepatotumorigenesis. METHODS: The expression of GGT in a chemically in- duced hepatocarcinogenesis model was examined by giving 0. 05% of 2-fluoenylacetamide in diet for 12 weeks. The ex- pression levels of total RNA and GGT, and the changes of liver pathology, GGT multiple molecular forms and sugar- chain heterogeneity were investigated at the different stages of rat hepatoma development. RESULTS: Pathological examination and biochemical ana- lysis found that liver GGT was over-expressed and secreted into blood during canceration. Serum total GGT and liver GGT specific activities (IU/g) including soluble and mem- brane-combined GGT were significantly higher (P <0.05) in experimental groups than those in control group, respec- tively. A highly positive correlation was found between to- tal GGT activities and total RNA levels (r =0.90, P <0.05) of the liver. Both were higher six weeks later than before. Con A-non-reactive-GGT was increased consistantly dur- ing the development of rat hepatoma. GGT multiple mo- lecular forms in the liver and sera of experimental rats showed that fetal liver-type GGT bands were associated with the development of hepatoma. CONCLUSIONS: Fetal liver-type GGT in sera and the liver of rats is closely related to hepatotumorigenesis. It can be used as a sensitive enzymatic marker for the early diagnosis of liver cancer.

关键词： gamma-glutamyltransferase multiple molecular forms rat hepatoma expression sugar-chain heterogeneity

Clinical impact of plasma TGF-β1 and circulating TGF-β1 mRNA in diagnosis of hepatocellular carcinoma

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Hepatobiliary & Pancreatic Diseases International 2008年第3期7卷 288-295页

作者： Zhi-Zhen Dong,Deng-Fu Yao,Min Yao,Li-Wei Qiu,Lei Zong,Wei Wu, Xin-Hua Wu,Deng-Bing Yao and Xian-Yong Meng Department of Diagnostics,Research Center of Clinical Molecular Biology,Department of Laboratory Science,and Department of GastroenterologyAffiliated Hospital and Institute of Neurosciences,nantong University,nantong 226001,china Department of Diagnostics Affiliated Hospital Nantong University Nantong 226001 China.

BACKGROUND:Transforming growth factor-β(TGF-β) plays an important role in the regulation of cell growth and differentiation,angiogenesis,extracellular matrix formation,immunosuppression and cancer development. In this study,we investigated the levels of TGF-β1 and TGF-β1 mRNA expression,their relationship with HBV replication,and their diagnostic value for hepatocellular carcinoma(HCC). METHODS:Total RNAs were extracted from HCC samples and matched non-tumor tissues,and from peripheral blood mononuclear cells in HCC ***-β1 mRNA was amplified by RT-PCR and confirmed by DNA sequencing. The distribution of TGF-β1 expression was assessed by *** clinical characteristics were analyzed between TGF-β1 and HBV *** diagnostic value of circulating TGF-β1 and TGF-β1 mRNA levels were investigated in HCC patients. RESULTS:The incidence of hepatic TGF-β1 expression was 83.3%in HCC samples,43.3%in the surrounding tissues, 94.7%in the HBV DNA-positive group,and 63.6%in the HBV DNA-negative *** TGF-β1 expression was associated with the degree of HCC differentiation and the status of HBV replication,but not with the size or number of *** TGF-β1 level and incidence of TGF-β1 mRNA were significantly higher in the HCC groupthan in any group of patients with benign liver disease, with a higher sensitivity of 89.5%and a specificity of 94.0% for HCC diagnosis when circulating TGF-β1 levels were >1.2μg/*** significant correlation was found between TGF-β1 expression and AFP level or tumor *** TGF-β1 level and serum AFP raised the detection rate to 97.4%. CONCLUSIONS:Abnormal expression of hepatic TGF-β1 is associated with the degree of HCC differentiation and HBV *** circulating TGF-β1 and TGF-β1 mRNA can be used as sensitive biomarkers for the diagnosis and prognosis of HBV-induced HCC.

关键词： hepatocellular carcinoma transforming growth factor beta immunohistochemistry diagnosis

Comparison of Hypolipidemic Effect of Refined Konjac Meal With Several Common Dietary Fibers and Their Mechanisms of Action

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Biomedical and Environmental Sciences 1997年第1期10卷 27-37页

作者： WU JIE AND PENG SHU-SHENG (Department of Nutrition and Food Hygiene, nantong Medical College, nantong City, 226001, Jiangsu, china Department of Nutrition and Food Hygiene, West china University of Medical Sciences,Chengdu, Sichuan, china) Department of Nutrition and Food Hygiene Nantong Medical College Jiangsu China.

The effects of RKM in comparison with pectin, algin and agar on lipid levels in serum and liver and on liver histopathology in rats were studied. In addition, the effects of all the tested materials on the composition and output of fecal bile acid were observed. All four kinds of dietary fiber were given at a level of 5% of diet to young male rats of Wistar strain fed on a lipid-rich diet contalning 5 % lard, 1% cholesteral and 0. 25 % cholate. All the dietary fibers tested have similar effects on serum lipid composition. In all groups, these substances prevent ed increases in total cholesterol in fasting serum, but the level of triglyceride was *** concentrations of totaI cholesterol and triglyceride in the liver were lower in the RKM group than in the control group and the other three groups. Hepatic histopathological exami nation also showed the most significant lipotropic effect in the RKM group. The daily output of fecal bile acids (CDCA+GDCA) was significantly increased in the four experimental groups than in the normal group and the control group. The increase of CDCA was more significant than GDCA, suggesting that the increase of fecal bile acids, especially CDCA, may be one of the mechanisms by which RKM and the other three dietary fibers exerts a hypocholesterolemic effect

关键词： acid Am Br Comparison of Hypolipidemic Effect of Refined Konjac Meal With Several Common Dietary Fibers and Their Mechanisms of Action CDCA

Protein expression profile in the differentiation of rat bone marrow stromal cells into Schwann cell-like cells

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Science china(Life Sciences) 2009年第3期52卷 267-277页

作者： LI WenTing1,SUN HuaLin2,XU ZengLu1,DING Fei2 & GU XiaoSong2 1 Department of Anatomy,Histology and Embryology,Peking Union Medical College,Institute of Basic Medical Science,Chinese Academy of Medical Sciences,Beijing 100005,china 2 Jiangsu Key Laboratory of Neuroregeneration,nantong University,nantong 226001,china 1. Department of Anatomy Histology and Embryology Peking Union Medical College Institute of Basic Medical Science Chinese Academy of Medical Sciences Beijing 100005 China 2. Jiangsu Key Laboratory of Neuroregeneration Nantong University Nantong 226001 China

During the last decade,increasing evidence suggested that bone marrow stromal cells(MSCs) have the potential to differentiate into neural *** studies have reported that MSCs showed morphological changes and expressed a limited number of neural proteins under experimental ***,no proteomic studies on MSCs differentiated into Schwann cell-like cells have been *** this study,we isolated MSCs from adult Sprague-Dawley rat femur and tibia bone marrows and induced the cells in vitro under specific *** using two-dimensional gel electrophoresis(2-DE),we compared the protein profiles of MSCs before and after induced *** obtained 792 protein spots in the protein profile by 2-DE,and found that 74 spots changed significantly before and after the differentiation using PDQuest software,with 43 up-regulated and 31 *** analyzed these 74 spots by a matrix assisted laser desorption ionization-time of flight mass spectrometry(MALDI-TOF-MS) and by database searching,and found that they could be grouped into various classes,including cytoskeleton and structure proteins,growth factors,metabolic proteins,chaperone proteins,receptor proteins,cell cycle proteins,calcium binding proteins,and other *** proteins also include neural and glial proteins,such as BDNF,CNTF and *** results may provide valuable proteomic information about the differentiation of MSCs into Schwann cell-like cells.

关键词： one marrow stromal cells(MSCs) differentiation Schwann cell-like cells proteomics

Quantitative analysis of vascular endothelial growth factor, microvascular density and their clinicopathologic features in human hepatocellular carcinoma

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Hepatobiliary & Pancreatic Diseases International 2005年第2期4卷 220-226页

作者： Deng-Fu Yao, Xin-Hua Wu, Yong Zhu, Gong-Sheng Shi, Zhi-Zhen Dong, Deng-Bing Yao, Wei Wu, Li-Wei Qiu and Xian-Yong Meng nantong, china Research Center of Clinical Molecular Biology , Department of Pathology and Department of Gastroenterology , Affiliated Hos- pital of nantong University Department of Diagnostics , and Institute of Neurosciences , nantong University nantong 226001, china Research Center of Clinical Molecular Biology Affiliated Hospital of Nantong University Nantong 226001 China. dfyrcmb@***

BACKGROUND: Angiogenesis is known to be essential to the survival, growth, invasion, and metastasis of tumor cells. Vascular endothelial growth factor (VEGF) are an important angiogenic factor regulating tumor angiogenesis, but its significance and tumor pathologic features are un- clear in hepatocellular carcinoma (HCC). In the present study, we analyzed expression of tissue VEGF, alteration of microvascular density (MVD) in microvessel angiogenesis, development and metastasis of HCC, and level of serum VEGF in differential diagnosis of benign and malignant liv- er diseases. METHODS: Tumor specimens were prospectively collected from HCC patients undergoing resection. Total RNAs were extracted and the expression levels were detected from different parts of HCC tissues. The cellular distributions of VEGF and MVD of liver tumors and their paracancerous and distal cancerous tissues were investigated by streptavi- din peroxidase (S-P) immunohistochemistry, respectively. The VEGF levels of circulating blood and hepatoma tissues were measured by enzyme-linked immunosorbent assay. RESULTS: The incidence of VEGF expression was 63.9% in HCCs (23/36 cases), 78.3% in non-encapsulated HCCs (18/23), and 90.9% in HCCs with extrahepatic metastasis (10/11), respectively. The VEGF expression was tightly correlated with MVD (P <0.01). The MVD in HCC with metastasis, low differentiation or non-encapsulation was significantly higher than that in HCC with intact capsule, high differentiation, or no metastasis. No significant diffe- rence was found between VEGF, MVD, tumor size, and hepatitis virus infection. The level of total RNA in HCC tis- sues was significantly lower but the VEGF level significantly higher than those in paracancerous or distal cancerous ones (P<0.01). The abnormal expression levels of VEGF in sera of HCC patients were directly correlated with the me- tastasis and recurrence of tumors. CONCLUSION: The high expression of VEGF and abnor- mality of tissue MVD are useful

关键词： vascular endothelial growth factor hepatocellular carcinoma ELISA microvascular density, immunohistochemistry

Abnormal expression of hepatomas and circulating telomerase and its clinical values

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Hepatobiliary & Pancreatic Diseases International 2005年第4期4卷 544-549页

作者： Wei Wu, Deng-Fu Yao, Li-Wei Qiu, Xin-Hua Wu, Ming Yao, Xiao-Qin Su and Li Zou nantong, china Research Center of Clinical Molecular Biology, Affiliated Hospital, nantong University, nantong 226001, china Research Center of Clinical Molecular Biology Affiliated Hospital Nantong University Nantong 226001 China.

BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignant tumours worldwide. Te-lomerase is reactivated in various types of malignant tumors and may contribute to the development of HCC. To evaluate the role of telomerase in formation and development of HCC, we analyzed its expression status in different parts of HCC tissues and peripheral blood mono-nuclear cells (PBMCs) , and explored its clinical implications for diagnosis of HCC. METHODS: Total RNAs and telomerase were extracted from HCC and their non-cancerous tissues, and both relationships were analyzed between them. The expression of telomerase reverse transcriptase (hTERT) mRNA and telomerase activities in liver tissues and PBMCs were detected by RT-PCR and telomeric repeat amplification protocol (TRAP)-ELISA, respectively. The diagnostic values of telomerase in PBMCs were investigated in the diagnosis and differentiation of HCC. RESULTS: The specific activities of telomerase were 18.25 ±15.02 A/μg RNA in HCC tissues, and significantly higher than those in their non-cancerous tissues (8.16 ± 6.22 A/μg RNA, P <0. 05). But total RNA levels in HCC were 12.40 ±7.34 μg/mg wet liver and lower than 53.77 ± 52.02 μg/mg wet liver in their non-cancerous parts (P < 0.01). The different telomerase levels could be detected in PBMCs from patients with chronic liver diseases and control group. The enzyme activities were significantly higher in HCC than in any other groups (P <0.01). However, circulating telomerase levels were more obviously decreased in HCC patients after transcatheter arterial embolization treatment than before the treatment (P <0.01). The analysis of combined serum alpha-fetoprotein level and PBMCs telomerase was more sensitive and specific for the diagnosis of HCC. CONCLUSION: The abnormal expression of telomerase in HCC tissues and circulating PBMCs could be a useful marker to the diagnosis and prognosis of HCC.

关键词： telomerase peripheral blood TRAP-ELISA hepatocellular carcinoma