AIM: To investigate the clinical values of serum free insulin-like growth factor Ⅱ (IGF-Ⅱ) levels and IGF-Ⅱ mRNA in hepatocellular carcinoma (HCC) tissues and peripheral blood for diagnosis of HCC and monitoring of...AIM: To investigate the clinical values of serum free insulin-like growth factor Ⅱ (IGF-Ⅱ) levels and IGF-Ⅱ mRNA in hepatocellular carcinoma (HCC) tissues and peripheral blood for diagnosis of HCC and monitoring of extrahepatic metastasis.METHODS: Total RNAs were extracted from HCC tissues or peripheral blood mononuclear cells from patients with HCC, liver diseases devoid of cancer, non-hepatic tumors,and healthy controls, respectively. IGF-Ⅱ cDNAs were synthesized through random primers and reversetranscriptase, amplified by polymerase chain reaction (PCR), and confirmed by DNA sequencing analysis. Serum free IGF-Ⅱ levels in patients with different liver diseases were analyzed by an enzyme-linked immunosorbent assay.RESULTS: The amplified fragments of IGF-Ⅱ mRNA by RT-PCR were identical to originally designed ones with a size of 170 bp and confirmed by sequencing analysis.The dilution experiments revealed that the lowest sensitivity of our system was 2 ng/L of total RNA. The positive frequencies of IGF-Ⅱ mRNA were 100% in HCC tissues,53.3% in para-cancerous tissues, and 0% in non-cancerous tissues, respectively. The serum free IGF-Ⅱ levels were significantly higher in HCC than those in chronic hepatitis or liver cirrhosis. The positive frequency of circulating IGF-Ⅱ mRNA was 34.2% in HCC, no amplified fragment was found in other liver diseases, extrahepatic tumors,and normal controls, respectively. The circulating IGF-Ⅱ mRNA correlated with the stage of HCC, and its positive rate was 100% in HCC with extrahepatic metastasis and 35.5% in HCC with AFP-negative. No significant correlation was found between tumor sizes and circulating IGF-Ⅱ mRNA fragment.CONCLUSION: The abnormal expressions of free IGF-Ⅱ and IGF-Ⅱ mRNA are useful tumor markers for HCC diagnosis, differentiation of extrahepatic metastasis and monitoring postoperative recurrence.展开更多
Micro-lens （ML） and Micro-lens array （MLA） are important optical components widely used in many fields; Soft-lithography, a vital little process technology, has its unique performance to produce ML and MLA; The cy...Micro-lens （ML） and Micro-lens array （MLA） are important optical components widely used in many fields; Soft-lithography, a vital little process technology, has its unique performance to produce ML and MLA; The cylinder and spherical MLA of polymethyl methacrylate （PMMA） were successfully obtained by micromolding inSoft-lithography. Some suitable experimental parameters in the process were discussed, and the imaging property of the MLA was also studied simply.展开更多
AIM: To investigate the effect of secreted frizzledrelated proteins(s FRPs) on CXC chemokine expression in human mesenchymal stem cells(h MSCs).METHODS: CXC chemokines such as CXCL5 and CXCL8 are induced in h MSCs dur...AIM: To investigate the effect of secreted frizzledrelated proteins(s FRPs) on CXC chemokine expression in human mesenchymal stem cells(h MSCs).METHODS: CXC chemokines such as CXCL5 and CXCL8 are induced in h MSCs during differentiation with osteogenic differentiation medium(OGM) and may be involved in angiogenic stimulation during bone repair. h MSCs were treated with conditioned medium(CM) from L-cells expressing non-canonical Wnt5 a protein, or with control CM from wild type L-cells, or directly with s FRPs for up to 10 d in culture. m RNA expression levels of both CXCL5 and CXCL8 were quantitated by real-time reverse transcriptase-polymerase chain reaction and secreted protein levels of these proteins determined by ELISA. Dose-(0-500 ng/m L) and time-response curves were generated for treatment with s FRP1. Signal transduction pathways were explored by western blot analysis with pan- or phosphorylation-specific antibodies, through use of specific pathway inhibitors, and through use of si RNAs targeting specific frizzled receptors(Fzd)-2 and 5 or thereceptor tyrosine kinase-like orphan receptor-2(Ro R2) prior to treatment with s FRPs. RESULTS: CM from L-cells expressing Wnt5 a, a noncanonical Wnt, stimulated an increase in CXCL5 m RNA expression and protein secretion in comparison to control L-cell CM. s FRP1, which should inhibit both canonical and non-canonical Wnt signaling, surprisingly enhanced the expression of CXCL5 at 7 and 10 d. Dickkopf1, an inhibitor of canonical Wnt signaling prevented the s FRPstimulated induction of CXCL5 and actually inhibited basal levels of CXCL5 expression at 7 but not at 10 d post treatment. In addition, all four s FRPs isoforms induced CXCL8 expression in a dose- and time-dependent manner with maximum expression at 7 d with treatment at 150 ng/m L. The largest increases in CXCL5 expression were seen from stimulation with s FRP1 or s FRP2. Analysis of mitogen-activated protein kinase signaling pathways in the presence of OGM showed s FRP1-induced phosphorylation of extracellular signal-regulated kinase(ERK)(p44/42) maximally at 5 min after s FRP1 addition, earlier than that found in OGM alone. Addition of a phospholipase C(PLC) inhibitor also prevented s FRPstimulated increases in CXCL8 m RNA. si RNA technology targeting the Fzd-2 and 5 and the non-canonical Fzd co-receptor Ro R2 also significantly decreased s FRP1/2-stimulated CXCL8 m RNA levels.CONCLUSION: CXC chemokine expression in h MSCs is controlled in part by s FRPs signaling through noncanonical Wnt involving Fzd2/5 and the ERK and PLC pathways.展开更多
An effective procedure was demonstrated to arrange spherical micro-beads into ordered, long, line-shape arrays by means of ＂micromolding in capillaries＂ in soft lithography. Polystyrene （PS） micro-beads with 2-3 m...An effective procedure was demonstrated to arrange spherical micro-beads into ordered, long, line-shape arrays by means of ＂micromolding in capillaries＂ in soft lithography. Polystyrene （PS） micro-beads with 2-3 mm of diameter were used as units and arranged by molding in continuous micro-channels formed by the conformal contact between a glass substrate and an elastomeric stamp with micrometer-scale line patterns on the surface. An aqueous emulsion of PS micro-beads filled these channels by capillary action and was allowed to solidify. The stamp was then removed. The PS micro-beads could be assembled into a string of long line-shape arrays, and the strings were then joined by heating them to their softening temperature. In order to separate the PS micro-bead string from the substrate, the glass was covered with a thin layer of A1 or polymethyl methacrylate. After the A1 layer was dissolved, the string of PS micro-beads would be released. A string of micrometer scale PS beads can be used as a simple and direct ＂model＂ of a real macromolecular chain. It is hopeful to show an analogue with the condensed process of real macromolecules in a mesoscopic scale using the ＂string of PS micro-beads＂.展开更多
BACKGROUND Systemic sclerosis is a rare connective tissue disease characterized by localized or diffuse skin thickening and fibrosis,which usually accumulates in various organs throughout the body.Tachyarrhythmia is a...BACKGROUND Systemic sclerosis is a rare connective tissue disease characterized by localized or diffuse skin thickening and fibrosis,which usually accumulates in various organs throughout the body.Tachyarrhythmia is a common clinical manifestation of cardiovascular damage in systemic sclerosis patients.However,few studies have reported the use of catheter ablation and an implantable cardioverter defibrillator in patients with systemic sclerosis complicated by ventricular tachycardia.CASE SUMMARY A 39-year woman with an 11-year history of systemic sclerosis was referred to our hospital due to three syncopal episodes in the past 6 mo.The results of an electrocardiogram and a transthoracic echocardiogram revealed ventricular tachycardia and left ventricular systolic and ventricular septum segmental motion abnormalities,respectively.The results of an electrocardiogram showed a sinus rhythm with complete blockage of the left bundle branch.In light of the progressive nature of systemic sclerosis,the presence of a left bundle branch block,and the decreased ejection fraction,a cardiac resynchronization therapydefibrillator was implanted.The patient’s clinical conditions improved,and at the3-mo follow-up,the patient was free of ventricular tachycardia and all cardiac symptoms.CONCLUSION We report the first case of systemic sderosis complicated by ventricular tachycardia that was successfully treated with a cardiac resynchronization therapy-defibrillator.展开更多
基金Supported by a grants-in-aid from the Key Project of Medical Science from Jiangsu Province, China, No. RC2003100
文摘AIM: To investigate the clinical values of serum free insulin-like growth factor Ⅱ (IGF-Ⅱ) levels and IGF-Ⅱ mRNA in hepatocellular carcinoma (HCC) tissues and peripheral blood for diagnosis of HCC and monitoring of extrahepatic metastasis.METHODS: Total RNAs were extracted from HCC tissues or peripheral blood mononuclear cells from patients with HCC, liver diseases devoid of cancer, non-hepatic tumors,and healthy controls, respectively. IGF-Ⅱ cDNAs were synthesized through random primers and reversetranscriptase, amplified by polymerase chain reaction (PCR), and confirmed by DNA sequencing analysis. Serum free IGF-Ⅱ levels in patients with different liver diseases were analyzed by an enzyme-linked immunosorbent assay.RESULTS: The amplified fragments of IGF-Ⅱ mRNA by RT-PCR were identical to originally designed ones with a size of 170 bp and confirmed by sequencing analysis.The dilution experiments revealed that the lowest sensitivity of our system was 2 ng/L of total RNA. The positive frequencies of IGF-Ⅱ mRNA were 100% in HCC tissues,53.3% in para-cancerous tissues, and 0% in non-cancerous tissues, respectively. The serum free IGF-Ⅱ levels were significantly higher in HCC than those in chronic hepatitis or liver cirrhosis. The positive frequency of circulating IGF-Ⅱ mRNA was 34.2% in HCC, no amplified fragment was found in other liver diseases, extrahepatic tumors,and normal controls, respectively. The circulating IGF-Ⅱ mRNA correlated with the stage of HCC, and its positive rate was 100% in HCC with extrahepatic metastasis and 35.5% in HCC with AFP-negative. No significant correlation was found between tumor sizes and circulating IGF-Ⅱ mRNA fragment.CONCLUSION: The abnormal expressions of free IGF-Ⅱ and IGF-Ⅱ mRNA are useful tumor markers for HCC diagnosis, differentiation of extrahepatic metastasis and monitoring postoperative recurrence.
基金Ⅶ. ACKN0WLEDGMENTS This work was supported by the National Natural Science Foundation of China （No.20374049） and the Specialized Research Fund for the Doctoral Program of Higher Education （No.20040358018）
文摘Micro-lens （ML） and Micro-lens array （MLA） are important optical components widely used in many fields; Soft-lithography, a vital little process technology, has its unique performance to produce ML and MLA; The cylinder and spherical MLA of polymethyl methacrylate （PMMA） were successfully obtained by micromolding inSoft-lithography. Some suitable experimental parameters in the process were discussed, and the imaging property of the MLA was also studied simply.
基金Supported by Merit Review Award from the United States,Department of Veterans Affairs Biomedical Laboratory Research and Development Service of the VA Office of Research and Development,No.I01BX000170
文摘AIM: To investigate the effect of secreted frizzledrelated proteins(s FRPs) on CXC chemokine expression in human mesenchymal stem cells(h MSCs).METHODS: CXC chemokines such as CXCL5 and CXCL8 are induced in h MSCs during differentiation with osteogenic differentiation medium(OGM) and may be involved in angiogenic stimulation during bone repair. h MSCs were treated with conditioned medium(CM) from L-cells expressing non-canonical Wnt5 a protein, or with control CM from wild type L-cells, or directly with s FRPs for up to 10 d in culture. m RNA expression levels of both CXCL5 and CXCL8 were quantitated by real-time reverse transcriptase-polymerase chain reaction and secreted protein levels of these proteins determined by ELISA. Dose-(0-500 ng/m L) and time-response curves were generated for treatment with s FRP1. Signal transduction pathways were explored by western blot analysis with pan- or phosphorylation-specific antibodies, through use of specific pathway inhibitors, and through use of si RNAs targeting specific frizzled receptors(Fzd)-2 and 5 or thereceptor tyrosine kinase-like orphan receptor-2(Ro R2) prior to treatment with s FRPs. RESULTS: CM from L-cells expressing Wnt5 a, a noncanonical Wnt, stimulated an increase in CXCL5 m RNA expression and protein secretion in comparison to control L-cell CM. s FRP1, which should inhibit both canonical and non-canonical Wnt signaling, surprisingly enhanced the expression of CXCL5 at 7 and 10 d. Dickkopf1, an inhibitor of canonical Wnt signaling prevented the s FRPstimulated induction of CXCL5 and actually inhibited basal levels of CXCL5 expression at 7 but not at 10 d post treatment. In addition, all four s FRPs isoforms induced CXCL8 expression in a dose- and time-dependent manner with maximum expression at 7 d with treatment at 150 ng/m L. The largest increases in CXCL5 expression were seen from stimulation with s FRP1 or s FRP2. Analysis of mitogen-activated protein kinase signaling pathways in the presence of OGM showed s FRP1-induced phosphorylation of extracellular signal-regulated kinase(ERK)(p44/42) maximally at 5 min after s FRP1 addition, earlier than that found in OGM alone. Addition of a phospholipase C(PLC) inhibitor also prevented s FRPstimulated increases in CXCL8 m RNA. si RNA technology targeting the Fzd-2 and 5 and the non-canonical Fzd co-receptor Ro R2 also significantly decreased s FRP1/2-stimulated CXCL8 m RNA levels.CONCLUSION: CXC chemokine expression in h MSCs is controlled in part by s FRPs signaling through noncanonical Wnt involving Fzd2/5 and the ERK and PLC pathways.
基金IV. ACKN0WLEDGMENT This work was supported by the National Natural Science Foundation of China （No.20374049） and the Specialized Research Foundation for the Doctoral Program of Higher Education （20040358018）.
文摘An effective procedure was demonstrated to arrange spherical micro-beads into ordered, long, line-shape arrays by means of ＂micromolding in capillaries＂ in soft lithography. Polystyrene （PS） micro-beads with 2-3 mm of diameter were used as units and arranged by molding in continuous micro-channels formed by the conformal contact between a glass substrate and an elastomeric stamp with micrometer-scale line patterns on the surface. An aqueous emulsion of PS micro-beads filled these channels by capillary action and was allowed to solidify. The stamp was then removed. The PS micro-beads could be assembled into a string of long line-shape arrays, and the strings were then joined by heating them to their softening temperature. In order to separate the PS micro-bead string from the substrate, the glass was covered with a thin layer of A1 or polymethyl methacrylate. After the A1 layer was dissolved, the string of PS micro-beads would be released. A string of micrometer scale PS beads can be used as a simple and direct ＂model＂ of a real macromolecular chain. It is hopeful to show an analogue with the condensed process of real macromolecules in a mesoscopic scale using the ＂string of PS micro-beads＂.
文摘BACKGROUND Systemic sclerosis is a rare connective tissue disease characterized by localized or diffuse skin thickening and fibrosis,which usually accumulates in various organs throughout the body.Tachyarrhythmia is a common clinical manifestation of cardiovascular damage in systemic sclerosis patients.However,few studies have reported the use of catheter ablation and an implantable cardioverter defibrillator in patients with systemic sclerosis complicated by ventricular tachycardia.CASE SUMMARY A 39-year woman with an 11-year history of systemic sclerosis was referred to our hospital due to three syncopal episodes in the past 6 mo.The results of an electrocardiogram and a transthoracic echocardiogram revealed ventricular tachycardia and left ventricular systolic and ventricular septum segmental motion abnormalities,respectively.The results of an electrocardiogram showed a sinus rhythm with complete blockage of the left bundle branch.In light of the progressive nature of systemic sclerosis,the presence of a left bundle branch block,and the decreased ejection fraction,a cardiac resynchronization therapydefibrillator was implanted.The patient’s clinical conditions improved,and at the3-mo follow-up,the patient was free of ventricular tachycardia and all cardiac symptoms.CONCLUSION We report the first case of systemic sderosis complicated by ventricular tachycardia that was successfully treated with a cardiac resynchronization therapy-defibrillator.
