AIM: To establish clone cells with different metastatic potential for the study of metastasis-related mechanisms. METHODS: Cloning procedure was performed on parental hepatocellular carcinoma (HCC) cell line MHCC97, a...AIM: To establish clone cells with different metastatic potential for the study of metastasis-related mechanisms. METHODS: Cloning procedure was performed on parental hepatocellular carcinoma (HCC) cell line MHCC97, and biological characteristics of the target clones selected by in vivo screening were studied. RESULTS: Two clones with high (MHCC97-H) and low (MHCC97-L) metastatic potential were isolated from the parent cell line. Compared with MHCC97-L, MHCC97-H had smaller cell size (average cell diameter 43 microm vs 50 microm) and faster in vitro and in vivo growth rate (tumor cell doubling time was 34.2h vs 60.0h). The main ranges of chromosomes were 55-58 in MHCC97-H and 57-62 in MHCC97-L. Boyden chamber in vitro invasion assay demonstrated that the number of penetrating cells through the artificial basement membrane was (37.5 +/- 11.0) cells/field for MHCC97-H vs (17.7 +/- 6.3)/field for MHCC97-L. The proportions of cells in G0-G1 phase, S phase, and G2-M phase for MHCC97-H/MHCC97-L were 0.56/0.65, 0.28/0.25 and 0.16/0.10, respectively, as measured by flow cytometry. The serum AFP levels in nude mice 5wk after orthotopic implantation of tumor tissue were (246 +/- 66) microg.L(-1) for MHCC97-H and (91 +/- 66) microg.L(-1) for MHCC97-L. The pulmonary metastatic rate was 100% (10/10) vs 40% (4/10). CONCLUSION: Two clones of the same genetic background but with different biological behaviors were established, which could be valuable models for investigation on HCC metastasis.展开更多
AIM: To determine whether normal genetically immunocompetent rodent hosts could be manipulated to accept human hepatocyte transplants with long term survival without immunosuppression. METHODS: Tolerance towards human...AIM: To determine whether normal genetically immunocompetent rodent hosts could be manipulated to accept human hepatocyte transplants with long term survival without immunosuppression. METHODS: Tolerance towards human hepatocytes was established by injection of primary human hepatocytes or Huh7 human hepatoma cells into the peritoneal cavities of fetal rats. Corresponding cells were subsequently transplanted into newborn rats via intrasplenic injection within 24h after birth. RESULTS: Mixed lymphocyte assays showed that spleen cells from non-tolerized rats were stimulated to proliferate when exposed to human hepatocytes, while cells from tolerized rats were not. Injections made between 15 d and 17 d of gestation produced optimal tolerization. Transplanted human hepatocytes in rat livers were visualized by immunohistochemical staining of human albumin. By dot blotting of genomic DNA in livers of tolerized rats 16 weeks after hepatocyte transplantation, it was found that approximately 2.5 X 10(5) human hepatocytes survived per rat liver. Human albumin mRNA was detected in rat livers by RT-PCR for 15 wk, and human albumin protein was also detectable in rat serum. CONCLUSION: Tolerization of an immuno-competent rat can permit transplantation, and survival of functional human hepatocytes.展开更多
目的研究糖尿病患者不同时段尿白蛋白（urinary albumin）的排泌情况及尿白蛋白在诊断早期糖尿病肾脏损伤中的应用。方法收集中山医院门诊及住院糖尿病患者及健康对照组3d内不同时间段的尿液，分析尿白蛋白天内、天间的排泌变化情况；以...目的研究糖尿病患者不同时段尿白蛋白（urinary albumin）的排泌情况及尿白蛋白在诊断早期糖尿病肾脏损伤中的应用。方法收集中山医院门诊及住院糖尿病患者及健康对照组3d内不同时间段的尿液，分析尿白蛋白天内、天间的排泌变化情况；以24h尿白蛋白为标准判断肾脏早期损伤情况，比较不同时段尿及时间点尿与24h尿白蛋白的相关性、诊断特异度及敏感度；评估随机尿的诊断特异度及敏感度，推导随机尿最佳诊断水平。结果尿白蛋白天间变异较大，以尿Cr和尿量分别校正后可降低变异。糖尿病组中尿白蛋白使用尿Cr校正后变异系数（CV小于尿量校正（CV分别为49％±23％vs 64％±30％）。尿白蛋白天内排泌呈节律性变化。不同尿液留取方式中夜间尿尿白蛋白／尿Cr（ratio of urinary concentrations of albumin and creatinine，ACR）与24h尿白蛋白定量相关性最好（R2=0．976），优于晨尿ACR（R2=0．900）、午间餐后尿ACR（R2=0．584）和随机尿ACR（R2=0．791）。以24h尿白蛋白总量作为判断标准进行受试者操作特性曲线（ROC曲线）分析显示，随机尿ACR的判断值为27．7μg／mg尿Cr（存在男女性别差异：男性12．8μg／mg尿Cr傩女性27．0μg／mg尿cr）。最小阴性似然比0．011时推导随机尿ACR的排除判断值为13．0μg／mg尿Cr；最大阳性似然比481．000时推导随机尿ACR的确诊判断值为87．4μg／mg尿Cr。结论尿Cr较尿量能更好地降低尿自蛋白天内变异，但仍无法完全消除变异。夜尿ACR与24h尿白蛋白定量相关性最好，可替代24h尿白蛋白定量。随机尿ACR作为最方便留取的尿液标本亦可以较好地替代24h尿白蛋白定量，但应考虑引入尿Cr后带来的性别间差异。以13．0μg／mg及87．4μg／mg作为随机尿ACR的排除判断值及确诊判断值可以便于临床医师基本排除或确定白蛋白尿的出现。展开更多
基金Supportod ty the State Key Basic Research Program Grant G1998051211 the Fund for Leading Specialty of Shanghai Metropolitan Bureau of Public Health.
文摘AIM: To establish clone cells with different metastatic potential for the study of metastasis-related mechanisms. METHODS: Cloning procedure was performed on parental hepatocellular carcinoma (HCC) cell line MHCC97, and biological characteristics of the target clones selected by in vivo screening were studied. RESULTS: Two clones with high (MHCC97-H) and low (MHCC97-L) metastatic potential were isolated from the parent cell line. Compared with MHCC97-L, MHCC97-H had smaller cell size (average cell diameter 43 microm vs 50 microm) and faster in vitro and in vivo growth rate (tumor cell doubling time was 34.2h vs 60.0h). The main ranges of chromosomes were 55-58 in MHCC97-H and 57-62 in MHCC97-L. Boyden chamber in vitro invasion assay demonstrated that the number of penetrating cells through the artificial basement membrane was (37.5 +/- 11.0) cells/field for MHCC97-H vs (17.7 +/- 6.3)/field for MHCC97-L. The proportions of cells in G0-G1 phase, S phase, and G2-M phase for MHCC97-H/MHCC97-L were 0.56/0.65, 0.28/0.25 and 0.16/0.10, respectively, as measured by flow cytometry. The serum AFP levels in nude mice 5wk after orthotopic implantation of tumor tissue were (246 +/- 66) microg.L(-1) for MHCC97-H and (91 +/- 66) microg.L(-1) for MHCC97-L. The pulmonary metastatic rate was 100% (10/10) vs 40% (4/10). CONCLUSION: Two clones of the same genetic background but with different biological behaviors were established, which could be valuable models for investigation on HCC metastasis.
文摘AIM: To determine whether normal genetically immunocompetent rodent hosts could be manipulated to accept human hepatocyte transplants with long term survival without immunosuppression. METHODS: Tolerance towards human hepatocytes was established by injection of primary human hepatocytes or Huh7 human hepatoma cells into the peritoneal cavities of fetal rats. Corresponding cells were subsequently transplanted into newborn rats via intrasplenic injection within 24h after birth. RESULTS: Mixed lymphocyte assays showed that spleen cells from non-tolerized rats were stimulated to proliferate when exposed to human hepatocytes, while cells from tolerized rats were not. Injections made between 15 d and 17 d of gestation produced optimal tolerization. Transplanted human hepatocytes in rat livers were visualized by immunohistochemical staining of human albumin. By dot blotting of genomic DNA in livers of tolerized rats 16 weeks after hepatocyte transplantation, it was found that approximately 2.5 X 10(5) human hepatocytes survived per rat liver. Human albumin mRNA was detected in rat livers by RT-PCR for 15 wk, and human albumin protein was also detectable in rat serum. CONCLUSION: Tolerization of an immuno-competent rat can permit transplantation, and survival of functional human hepatocytes.
文摘目的研究糖尿病患者不同时段尿白蛋白（urinary albumin）的排泌情况及尿白蛋白在诊断早期糖尿病肾脏损伤中的应用。方法收集中山医院门诊及住院糖尿病患者及健康对照组3d内不同时间段的尿液，分析尿白蛋白天内、天间的排泌变化情况；以24h尿白蛋白为标准判断肾脏早期损伤情况，比较不同时段尿及时间点尿与24h尿白蛋白的相关性、诊断特异度及敏感度；评估随机尿的诊断特异度及敏感度，推导随机尿最佳诊断水平。结果尿白蛋白天间变异较大，以尿Cr和尿量分别校正后可降低变异。糖尿病组中尿白蛋白使用尿Cr校正后变异系数（CV小于尿量校正（CV分别为49％±23％vs 64％±30％）。尿白蛋白天内排泌呈节律性变化。不同尿液留取方式中夜间尿尿白蛋白／尿Cr（ratio of urinary concentrations of albumin and creatinine，ACR）与24h尿白蛋白定量相关性最好（R2=0．976），优于晨尿ACR（R2=0．900）、午间餐后尿ACR（R2=0．584）和随机尿ACR（R2=0．791）。以24h尿白蛋白总量作为判断标准进行受试者操作特性曲线（ROC曲线）分析显示，随机尿ACR的判断值为27．7μg／mg尿Cr（存在男女性别差异：男性12．8μg／mg尿Cr傩女性27．0μg／mg尿cr）。最小阴性似然比0．011时推导随机尿ACR的排除判断值为13．0μg／mg尿Cr；最大阳性似然比481．000时推导随机尿ACR的确诊判断值为87．4μg／mg尿Cr。结论尿Cr较尿量能更好地降低尿自蛋白天内变异，但仍无法完全消除变异。夜尿ACR与24h尿白蛋白定量相关性最好，可替代24h尿白蛋白定量。随机尿ACR作为最方便留取的尿液标本亦可以较好地替代24h尿白蛋白定量，但应考虑引入尿Cr后带来的性别间差异。以13．0μg／mg及87．4μg／mg作为随机尿ACR的排除判断值及确诊判断值可以便于临床医师基本排除或确定白蛋白尿的出现。
