目的探讨肝癌细胞在低渗溶液中的存活状态,为临床应用温热蒸馏水杀灭腹腔游离肝癌细胞提供理论依据。方法对体外培养的肝癌细胞株Hep3B分别应用渗透压为148 m Osmol/kg和90 m Osmol/kg的羟乙基哌嗪乙硫磺酸(HEPES)缓冲溶液及0 m Osmol/k...目的探讨肝癌细胞在低渗溶液中的存活状态,为临床应用温热蒸馏水杀灭腹腔游离肝癌细胞提供理论依据。方法对体外培养的肝癌细胞株Hep3B分别应用渗透压为148 m Osmol/kg和90 m Osmol/kg的羟乙基哌嗪乙硫磺酸(HEPES)缓冲溶液及0 m Osmol/kg的蒸馏水作用后,运用MTT法及流式细胞仪观察其活性及状态。结果 148 m Osmol/kg及90 m Osmol/kg渗透压的HEPES缓冲溶液对肝癌细胞株Hep3B的杀伤作用不明显,作用30 min后仍有较强的活性;0 m Osmol/kg渗透压的蒸馏水对肝癌细胞有一定的杀伤作用,且随作用时间的延长其杀伤效果增强,作用10 min后才可杀灭肿瘤细胞。结论在肝癌手术中应用温热蒸馏水灌洗需要持续作用10 min以上方可起到杀灭腹腔内游离癌细胞的作用,进而预防术后腹膜种植转移的发生。展开更多
BACKGROUND: Aquaporin-4 (AQP4) is abundant in astrocytes, ependymal cells, and the choroid plexus, and is associated with cerebrospinal fluid formation and osmoregulation. AQP4 is speculated to be the hypothalamic ...BACKGROUND: Aquaporin-4 (AQP4) is abundant in astrocytes, ependymal cells, and the choroid plexus, and is associated with cerebrospinal fluid formation and osmoregulation. AQP4 is speculated to be the hypothalamic osmoreceptor and regulator of water balance. OBJECTIVE: To examine AQP4 expression and its role in cultured rat astrocytes after exposure to hypotonic medium. DESIGN, TIME AND SETTING: Randomized control experiment. This experiment was carried out in the Research Room of Neurobiology, Chongqing University of Medical Science, China, between April and October 2003. MATERIALS: Two-day-old newborn Wistar rats (n =20), weighing 10- 15 g, were purchased from the Experimental Animal Center of Chongqing University of Medical Science, China. METHODS: Purified rat cerebral cortical astrocytes were isolated fiom Wistar rats for in vitro cell culture experiments. The cells were randomly divided into control and hypotonic groups. The in vitro cell edema model was established by exposing astrocytes to hypotonic medium (268, 254, or 240 mmol/L). Cells in the control group were cultured in normal culture medium. MAIN OUTCOME MEASURES: Morphological changes in astrocytes were observed under an inverted microscope and a transmission electron microscope after cells were cultured for 3, 6, 12, or 24 hours with hypotonic medium or normal culture medium. In each group, AQP4 protein and mRNA expression were assessed by immunocytochemistry, in situ hybridization, and reverse transcription polymerase chain reaction at the different time points. RESULTS: After astrocytes were cultured for 3, 6, 12, or 24 hours with hypotonic medium (268, 254, 240 mmol/L), they showed typical features of cell edema. In the control group, no astrocytes developed pathological changes. There were no significant changes in the AQP4 mRNA and protein expression in the control group at any of the time points alter astrocytes were cultured with normal culture medium (P 〉 0.05). Compared with the control group, AQP4 mRN展开更多
文摘目的探讨肝癌细胞在低渗溶液中的存活状态,为临床应用温热蒸馏水杀灭腹腔游离肝癌细胞提供理论依据。方法对体外培养的肝癌细胞株Hep3B分别应用渗透压为148 m Osmol/kg和90 m Osmol/kg的羟乙基哌嗪乙硫磺酸(HEPES)缓冲溶液及0 m Osmol/kg的蒸馏水作用后,运用MTT法及流式细胞仪观察其活性及状态。结果 148 m Osmol/kg及90 m Osmol/kg渗透压的HEPES缓冲溶液对肝癌细胞株Hep3B的杀伤作用不明显,作用30 min后仍有较强的活性;0 m Osmol/kg渗透压的蒸馏水对肝癌细胞有一定的杀伤作用,且随作用时间的延长其杀伤效果增强,作用10 min后才可杀灭肿瘤细胞。结论在肝癌手术中应用温热蒸馏水灌洗需要持续作用10 min以上方可起到杀灭腹腔内游离癌细胞的作用,进而预防术后腹膜种植转移的发生。
基金the National Natural Science Foundation of China, No.30070247
文摘BACKGROUND: Aquaporin-4 (AQP4) is abundant in astrocytes, ependymal cells, and the choroid plexus, and is associated with cerebrospinal fluid formation and osmoregulation. AQP4 is speculated to be the hypothalamic osmoreceptor and regulator of water balance. OBJECTIVE: To examine AQP4 expression and its role in cultured rat astrocytes after exposure to hypotonic medium. DESIGN, TIME AND SETTING: Randomized control experiment. This experiment was carried out in the Research Room of Neurobiology, Chongqing University of Medical Science, China, between April and October 2003. MATERIALS: Two-day-old newborn Wistar rats (n =20), weighing 10- 15 g, were purchased from the Experimental Animal Center of Chongqing University of Medical Science, China. METHODS: Purified rat cerebral cortical astrocytes were isolated fiom Wistar rats for in vitro cell culture experiments. The cells were randomly divided into control and hypotonic groups. The in vitro cell edema model was established by exposing astrocytes to hypotonic medium (268, 254, or 240 mmol/L). Cells in the control group were cultured in normal culture medium. MAIN OUTCOME MEASURES: Morphological changes in astrocytes were observed under an inverted microscope and a transmission electron microscope after cells were cultured for 3, 6, 12, or 24 hours with hypotonic medium or normal culture medium. In each group, AQP4 protein and mRNA expression were assessed by immunocytochemistry, in situ hybridization, and reverse transcription polymerase chain reaction at the different time points. RESULTS: After astrocytes were cultured for 3, 6, 12, or 24 hours with hypotonic medium (268, 254, 240 mmol/L), they showed typical features of cell edema. In the control group, no astrocytes developed pathological changes. There were no significant changes in the AQP4 mRNA and protein expression in the control group at any of the time points alter astrocytes were cultured with normal culture medium (P 〉 0.05). Compared with the control group, AQP4 mRN
基金supported by the National Natural Science Foundation of China(No.3077110630870567+2 种基金30871267)Natura Science Foundation of Guangdong ProvinceChina(No.7005974)