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

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

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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

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Abnormal expression of HSP gp96 associated with HBV replication in human hepatocellular carcinoma

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Hepatobiliary & Pancreatic Diseases International 2006年第3期5卷 381-386页

作者： Deng-Fu Yao,Xin-Hua Wu,Xiao-Qin Su,Min Yao, Wei Wu, Li-Wei Qiu, Li Zou and Xian-Yong Meng The Institute of Clinical Medicine, nantong university, and department of Gastroenterology,Affiliated Hospital of nantong university, nantong 226001, china and Nanjing Medical university, Nanjing 210009, china The Institute of Clinical Medicine Nantong University Nantong 226001 China. yaodf@***

BACKGROUND: Heat shock protein (HSP) gp96 is a member of the HSP90 family and presumably overexpresses as a result of stimulation by mutated or abnormal proteins. Its abnormal expression correlates with carcinogenesis, progression and prognosis of hepatocellular carcinoma (HCC). In this study, we investigated the pathological characteristics of liver gp96 expression and its relationship with hepatitis B virus (HBV) replication in HCC patients. METHODS: Tumor specimens were prospectively collected from 30 HCC patients undergoing liver resection. Total RNAs were extracted from HCC or their noncancerous tissues. The distribution of gp96 expression in hepatocytes was investigated by streptavidin peroxidase (S-P) immunohistochemistry and tissue HBV-DNA was detected by the in situ molecular hybridization technique. The association of gp96 expression with HBV replication, and the histopathological characteristics of HCC were analyzed. RESULTS: The gp96 was strongly expressed in HCC (73.3%, 22 of 30) and weakly (46.7%, 14 of 30) in non-cancerous tissues. The gp96 expression in HCC tissues was correlated with degree of tumor differentiation and tumor size, but not with tumor number (P>0.05). Immunohistochemical analysis showed that 17 of 19 HCC patients with HBVDNA -positive were strongly expressed for gp96, whereas only 5 of 11 patients with HBV-DNA-negative were positive for gp96. A significant difference was found between the two groups (89.5% vs. 45.5%, P<0.05). CONCLUSIONS: The abnormal expressions of HSP gp96 in HCC tissues are associated with HBV replication. This finding indicates that HBV infection plays an important role in the development of HCC.

关键词： hepatocellular carcinoma heat shock protein gp96 immunohistochemistry HBV-replication

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Surgical repair of a 30-mm-long human median nerve defect in the distal forearm by implantation of a chitosan/PGA nerve guidance conduit:a case study

Surgical repair of a 30-mm-long human median nerve defect in...

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中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Wen HU~1,Jian-Hui GU~2,Ai-Dong DENG~2,Xiao-Song GU~(1,*) 1 jiangsu Key Laboratory of Neuroregeneration,nantong university,nantong,226001 2 department of Hand Surgery,Affiliated Hospital of nantong university,nantong,226001

Reconstruction of injured peripheral nerve by bridging neural defects with artificial nerve guidance conduits (NGCs),as an alternative to autologous nerve grafts,becomes one of the main concerns in the fields of microsurgery, plastic surgery,and regenerative *** have developed a novel design of artificial NGCs consisting of a chitosan conduit and longitudinally aligned filaments of polyglycolic acid(PGA) inserted in the conduit *** chitosan/PGA NGCs showed good biological properties,and yielded promising repair outcomes after they were used to bridge canine sciatic nerve defects as long as 30 *** also reported a case study in which a 35-mm-long human median nerve defect at elbow was repaired by the chitosan/PGA *** we describe another clinical case study in which a chitosan/PGA NGC was utilized to repair a 30-mm-long median nerve defect in the distal forearm of a 55-year-old ***-six months post-implantation,the function of the right abductor pollicis and opponens pollicis recovered to an M4 level. Static two-point discrimination measured 14,9,and 9 mm in the thumb,index and middle fingers of the right hand, indicating a S3 level sensory recovery for the injured median *** compound muscle action potentials were recorded on the right abductor *** ninnydrin test showed partial recovery in perspiration of the injured thumb,index and middle *** repair case suggests a potential strategy for clinical reconstruction of extended defects in peripheral nerve trunks by implantation of chitosan/PGA NGCs.

关键词： nerve injury peripheral nerve surgery median nerve nerve guidance conduits chitosan polyglycolic acid

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The Effect of Anti-human CD134 Monoclonal Antibody on Phytohemagglutinin-Induced mRNA Expression of Perforin in Peripheral Blood Mononuclear Cells

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Cellular & Molecular Immunology 2005年第6期2卷 467-471页

作者： Ming Li~1 Yuanchao Zhang~(1,2) ~1department of Rheumatology and Immunology,Shandong Provincial Hospital,Shandong university,Jinan 250021,china ~2department of Rheumatology and Immunology,Shandong Provincial Hospital,Shandong university,Jinan 250021,Shandong,china. department of Rheumatology and Immunology Shandong Provincial Hospital Shandong University Jinan 250021 China.

CD134, a member of the tumor necrosis factor receptor superfamily, plays a crucial role in T cell survival. In this study, peripheral blood mononuclear cells （PBMCs） stimulated with phytohemagglutinin （PHA, 50 μg/ml） were treated by CD134 mAb （1μg/ml, 5 μg/ml, 10 μg/ml） for 6 h, 12 h, 24 h and 48 h. The level of perforin mRNA was measured by reverse transcription-polymerase chain reaction （RT-PCR） technique. Our data showed that the expression of perforin mRNA in PBMCs was down-regulated by CD134 mAb in a dose-dependent manner in range of 1 μg/ml to 5 μg/ml and dropped down to its minimum on 24 h （p 〈 0.05）. The level of perforin mRNA reached a plateau when the concentration of CD134 mAb exceeded 5 μg/ml. In conclusion, CD134 mAb can inhibit perforin expression, which may enhance the ability of T cells for survival. Cellular ＆ Molecular Immunology. 2005;2（6）： 467-471.

关键词： CD134 perforin phytohemagglutinin

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NET-1和MIB-1在肝癌中的表达及意义

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天津医药 2009年第11期37卷 917-919,I0001页

作者：李杏玉陈莉江苏省南通大学医学院病理解剖学教研室 226001

目的:研究NET-1和MIB-1在肝细胞肝癌(HCC)中的表达意义,分析两者与HCC生物学行为的相关性。方法:采用间接免疫荧光细胞化学法结合激光扫描共聚焦显微镜检测肝癌细胞株SMMC-7721细胞中NET-1与MIB-1的定位表达;同时以免疫组织化学SP法和... 详细信息

目的:研究NET-1和MIB-1在肝细胞肝癌(HCC)中的表达意义,分析两者与HCC生物学行为的相关性。方法:采用间接免疫荧光细胞化学法结合激光扫描共聚焦显微镜检测肝癌细胞株SMMC-7721细胞中NET-1与MIB-1的定位表达;同时以免疫组织化学SP法和双标双染(DouS-P)法检测76例肝癌组织标本中NET-1与MIB-1的表达。结果:NET-1表达于肝癌细胞胞浆,MIB-1表达于肝癌细胞胞核。HCC中NET-1与MIB-1强阳性表达率分别为51.32%和38.16%,两指标表达呈正相关(rs=0.3342,P<0.01)。NET-1、MIB-1表达强度在不同HCC肿瘤病理分级、临床分期和是否伴有肿瘤淋巴结转移患者中差异均有统计学意义(P<0.05或P<0.01)。另外,不同HCC细胞学类型以及是否伴有肝炎肝硬化患者中,NET-1表达差异也具有统计学意义(P<0.01)。结论:NET-1作为胞浆蛋白在HCC中过表达,与MIB-1表达密切相关。提示NET-1为肿瘤增殖相关蛋白,在肿瘤中显著上调,并与HCC恶性生物学行为相关。

关键词：肝肿瘤 NET-1 Ki-67抗原肿瘤细胞培养的癌

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Mitochondrial uncoupling protein 2 expression in colon cancer and its clinical significance

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World Journal of Gastroenterology 2010年第45期16卷 5773-5778页

作者： Xiao-Yi Kuai, Ze-Yu Ji, Hong-Jie Zhang,department of Gastroenterology, First Affiliated Hospital of Nanjing Medical university, Nanjing 210029, jiangsu province, china department of Gastroenterology First Affiliated Hospital of Nanjing Medical University 300 Guangzhou Road Nanjing 210029 Jiangsu Province China. hjzhang06@***

AIM: To detect the expression of mitochondrial uncoupling protein 2 (UCP2) in colon cancer and analyze the relation between UCP2 expression and clinical pathological features of colon ***: Fifteen colon tissue samples and 15 its adjacent tissue samples were obtained from colon cancer patients during surgical interventions. UCP2 expression was detected with immunohistochemical method in 10 normal controls, 10 hyperplastic polyp patients, 20 tubular adenoma patients and 78 colon cancer patients. Patients with rectal cancer were excluded. Quantitative reverse transcription polymerase chain reaction and Western blotting were used to detect UCP2 expressions in colon cancer tissue samples and its adjacent tissue samples. Relation between UCP2 expression and clinical pathological features of colon cancer was also analyzed. RESULTS: The UCP2 mRNA expression level was fourfold higher in colon cancer tissue samples than in its adjacent tissue samples. The UCP2 protein expression level was three-fold higher in colon cancer tissue samples than in its adjacent normal tissue samples. The UCP2 was mainly expressed in cytoplasm. The UCP2 was not expressed in normal colon mucosa. Strong positive staining for UCP2 with a diffuse distribution pattern was identified throughout the mucosa in colon cancer tissue samples with a positive expression rate of 85.9%. The UCP2 expression level was higher in colon cancer tissue samples at clinical stages Ⅲ and Ⅳ than in those at stageⅠ+ Ⅱ. Univariate analysis showed that the high UCP2 expression level was significantly correlated to colon cancer metastasis (hazard ratio = 4.321, confidence interval = 0.035-0.682, P = 0.046). CONCLUSION: UCP2 is highly expressed in human colon cancer tissue and may be involved in colon cancer metastasis.

关键词： Mitochondrial uncoupling protein 2 Colon cancer Uncoupling protein 2 Clinicopathologic characteristics

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ERK1/2 contributes negative regulation to STAT3 activity in HSS-transfected HepG2 cells

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Cell Research 2004年第2期14卷 141-147页

作者： Ze Jun TIAN~2, Wei AN~(1,*) ~1department of Cell Biology, Capital university of Medical Science, 10 Xitoutiao, You An Men Beijing 100054, china. ~2department of Cardiology, Second Hospital, Hebei Medical university, Shijiazhuang, china.. 2department of Cardiology Second Hospital Hebei Medical University Shijiazhuang China.. 1department of Cell Biology Capital University of Medical Science 10 Xitoutiao You An Men Beijing 100054 China.

Signal transducer and activator of transcription 3 (STAT3) is a recently characterized transcription factor which is essential to liver regeneration. We have previously reported that hepatic stimulator substance (HSS), a novel growthpromoting substance, phosphorylated the epidermal growth factor (EGF) receptors and activated downstream Ras-MAP kinase (extracellular signal-regulated kinases, ERK1/2) cascade. However, whether HSS signal is related to STAT3 pathway remains unclear. The present study is aiming to explore the regulatory effect of activation of ERK1/2 evoked by HSS on STAT3 phosphorylation and STAT3 signaling. human hepatoma cell line HepG2 was stably transfected with HSS cDNA and HSS expression was measured by Northern blot. The results showed that the transfection of HSS into HepG2 resulted in remarkable increase in cellular proliferation as compared with the non-transfected cells, and it was further proved that the cellular proliferation in the HSS-transfected cells was related to ERK1/2 a

关键词： hepatic stimulator substance ERK1/2 STAT3 hepatocyte growth.

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Artificial nerve grafts sustain regenerated axons from prolonged axotomized neurons to chronically denervated segment

Artificial nerve grafts sustain regenerated axons from prolo...

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中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Jian YAO~1,Hai-Shan JIAO~1,Xue CHEN~1,Wei-Wei LIN~1,Yi LI~1,Yu-Ming YANG~2,Xiao-Song GU~2,Xiao-Dong WANG~(1,2,*) 1 department of Histology and Embryology,Medical College,nantong university,nantong,226001,china 2 jiangsu Key Laboratory of Neuroregeneration,nantong university,nantong,226001,china

＜正＞Objective Peripheral nerve regeneration for long-term delayed injuries is usually *** we attempted to use a chitosan/polyglycolic acid（PGA） artificial nerve graft to bridge a long-term delayed 10-mm defect in SD rats based on the previous studies on the graft used for immediate repair of 30-mm long dog sciatic nerve defects and for clinical treatment of a 35-mm-long median nerve defect at elbow of a human *** In this study,for experimental groups,the rat sciatic nerve had been transected leaving a 10-mm defect,which was maintained for 3 or 6 months before implantation with the chitosan/PGA artificial nerve *** animals non-grafted or grafted with autograft served as negative or positive control *** experiment groups,nerve regeneration with functional recovery were achieved as measured by electrophysiological and histological techniques,although differences in the quantity and the quality of the regenerated nerve were observed between the 3- and 6-month delayed ***（1） A few denervated Schwann cells survived and sustained their ability to myelinate axons at least 6 months,and（2） the atrophic denervated muscle could be reinnervated by regenerated axons through new muscle-nerve *** These observations provide the possibility of guiding regenerated axons from survived axotomized neurons to distal nerve stump by the chitosan/PGA artificial nerve graft.

关键词： nerve tissue engineering nerve regeneration long-term axotomy chronic denervation artiticial nerve gratt reinnervation

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Overexpression of heme oxygenasel protects smooth muscle cells against oxidative injury and inhibits cell proliferation

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Cell Research 2002年第2期12卷 123-132页

作者： MIN ZHANG, BAO HuI ZHANG, LI CHEN, WEI AN1 Institute of Sports Medicine, The Third Hospital, Peking university, Beijing 100083, china 2department of Cell Biology, Capital university of Medical Sciences, Beijing 100054, china 1Institute of Sports Medicine The Third Hospital Peking University Beijing 100083 China 1Institute of Sports Medicine The Third Hospital Peking University Beijing 100083 China 2department of Cell Biology Capital University of Medical Sciences Beijing 100054 China 2department of Cell Biology Capital University of Medical Sciences Beijing 100054 China

To investigate whether the expression of exogenous heme oxygenase-1 (HO-1) gene within vascular smooth muscle cells (VSMC) could protect the cells from free radical attack and inhibit cell proliferation, we established an in vitro transfection of human HO-1 gene into rat VSMC mediated by a retroviral vector. The results showed that the profound expression of HO-1 protein as well as HO activity was 1.8- and 2.0-fold increased respectively in the transfected cells compared to the non-transfected ones. The treatment of VSMC with different concentrations of H2O2 led to the remarkable cell damage as indicated by survival rate and LDH leakage. However, the resistance of the HO-1 transfected VSMC against H2O2 was significantly raised. This protective effect was dramatically diminished when the transfected VSMC were pretreated with ZnPP-IX, a specific inhibitor of HO, for 24 h. In addition, we found that the growth potential of the transfected cells was significantly inhibited directly by increased activity of H

关键词： Heme oxygenase retroviral vector free radical cell proliferation vascular smooth muscle cells.

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