BACKGROUND: Cryoanalgesia at -50 ℃ for 90 seconds yields effective pain relief followingthoracotomy. In China, -50 ℃ is a common temperature for intercostal cryoanalgesia followingthoracotomy. However, experimental ...BACKGROUND: Cryoanalgesia at -50 ℃ for 90 seconds yields effective pain relief followingthoracotomy. In China, -50 ℃ is a common temperature for intercostal cryoanalgesia followingthoracotomy. However, experimental results vary.OBJECTIVE: To explore intercostal nerve pathological changes at-70 ℃ for various freezing timesby studying canines, and to evaluate long-term clinical efficacy of intercostal nerve cryoanalgesia forpostoperative pain relief based on the animal experiments.DESIGN, TIME AND SETTING: A comparative animal study was performed at the AnimalExperimental Center of the General Hospital of the People's Liberation Army. Based on results fromthe animal study, a randomized, controlled, clinical trial was performed at the Department ofThoracic Surgery of the General Hospital of the People's Liberation Army between October 2006and October 2008.PARTICIPANTS: A total of 120 patients undergoing posterolateral single incision Iobectomy at theDepartment of Thoracic Surgery of PLA General Hospital between October 2006 and October 2008were selected. Nervous system diseases were excluded.METHODS: Animal experiment: 8 anaesthetized, mixed-breed dogs were used. The intercostalnerves (costal bone 6-10) were frozen at -70 ℃ for varying times (30, 60, 90, 120, and 180seconds). Clinical study: 120 patients were randomly assigned to 2 groups (n = 60). In thecryoanalgesia group, the intercostal nerves were frozen prior to chest closure, and 4 costal nerves(1 at incision level, 2 above and below incision, and 1 at drainage tube level) were frozen for 90seconds at -70 ℃, respectively. Intercostal nerves were not frozen in the control group patients.Dolantin was used to relieve postoperative pain in patients from both groups.MAIN OUTCOME MEASURES: Pathological changes in frozen intercostal nerves were examined atdays 1, 10, 30, and 60 following freezing. Following surgery, the degree of postoperative pain in allpatients was evaluated by visual analogue scale at days 1,3, 5, 9, 30, 60, 90, and 180. Dolantindoses at days 1, 3, 5, 9 post-surgery and postoperative complications were noted.RESULTS: Nerve damage progressively increased with length of freezing time at-70 ℃, andrecovery time from damage was gradually increased. After freezing for 90 seconds, the nervesexhibited obvious histopathological damage, and then completely recovered. In addition, afterfreezing for 180 seconds, the histopathological changes in nerves were reversible. In the clinicalstudy, visual analogue scale scores were significantly less in the cryoanalgesia group compared withthe control group (P < 0.01), which was maintained over 30 days. In the cryoanalgesia group, themean dolantin dose administered and postoperative complications were significantly reducedcompared with the control group (P< 0.01).CONCLUSION: Freezing of the intercostal nerve at -70 ℃ for 90 seconds is a safe and long-termeffective method for relieving post-thoracotomy pain.展开更多
BACKGROUND: Dendritic cell is the most major antigen presenting cell of organism. It is proved in recent studies that human umbilical cord blood mononuclear cells induced and cultured in vitro by recombinant human gra...BACKGROUND: Dendritic cell is the most major antigen presenting cell of organism. It is proved in recent studies that human umbilical cord blood mononuclear cells induced and cultured in vitro by recombinant human granulocyte-macrophage colony stimulating factor (rhG-MCSF) and recombinant human interleukin-4(rhIL-4) can generate a great many dendritic cells and promote the lethal effect of T cells on human neuroblastoma, but it is unclear that whether the lethal effect is associated with the most proper concentration of dendritic cells. OBJECTIVE: To investigate the lethal effect of human umbilical cord blood mononuclear cells induced in vitro by cytokines differentiating into dendritic cells on human neuroblastoma, and its best concentration range. DESIGN: Open experiment. SETTING: Department of Pediatrics, the Medical School Hospital of Qingdao University. MATERIALS: The study was carried out in the Shandong Provincial Key Laboratory (Laboratory for the Department of Pediatrics of the Medical School Hospital of Qingdao University) during September 2005 to May 2006. Human umbilical cord blood samples were taken from the healthy newborn infants of full-term normal delivery during October to November 2005 in the Medical School Hospital of Qingdao University, and were voluntarily donated by the puerperas. Main instruments: type 3111 CO2 incubator (Forma Scientific, USA), type 550 ELISA Reader (Bio-Rad, USA). Main reagents: neuroblastoma cell line SK-N-SH (Shanghai Institute of Life Science, Chinese Academy of Sciences), RPMI-1640 culture fluid and fetal bovine serum (Hyclone), rhIL-4 (Promega, USA), rhG-MCSF (Harbin Pharmaceutic Group Bioengineering Co.Ltd), rat anti-human CD1a monoclonal antibody and FITC-labeled rabbit anti-rat IgG (Xiehe Stem cell Gene Engineering Co.Ltd). METHODS: ① Human umbilical cord blood mononuclear cells obtained with attachment methods differentiated into human umbilical cord blood dendritic cells, presenting typical morphology of dendritic cells after in vitro induction by rhG-MCSF and rhIL-4. ② Different concentrations of dendritic cells[ dendritic cells: neuroblastoma cells=20∶1,50∶1,100∶1(2×108 L-1,5×108 L-1,1×109 L-1)], 1×109 L-1 T cells and 1×107 L-1 neuroblastoma cells were added in the experimental group. 1×109 L-1 T cells and 1×107 L-1 neuroblastoma cells were added in the control group. ③ Main surface marker CD1a molecules of dendritic cells were detected with indirect immunofluorescence, and the percent rate of dendritic cells was counted with ultraviolet light and expressed as the expression rate of CD1a+ cells. ④ Single effector cells and target cells were respectively set in the experimental group and control group to obtain the lethal effect. The lethal effect of dendritic cells on neuroblastoma cells was indirectly evaluated by detecting cellular survival with MTT assay. The lethal effect(%)=(1-A experimental well-A effector cell well/A target cell well)×100%.⑤The experimental data were presented as Mean ±SD, and paired t test was used. MAIN OUTCOME MEASURES: ① Morphological characters of dendritic cells in the process of induction and differentiation. ②CD1a+ cellular expression rate. ③Lethal effect of dendritic cells on neuroblastoma cells. RESULTS: ①Morphological characters of dendritic cells in the process of induction and differentiation: On the 15th day after human umbilical cord blood mononuclear cells were induced by rhG-MCSF and rhIL-4, typical morphology of dendritic cells could be seen under an inverted microscope. ②Expression rate of CD1a+ cells was (43.12±5.83)%. ③Lethal effect of dendritic cells on neuroblastoma cells: Lethal effect of dendritic cells stimulated T cells in each experimental group ( dendritic cells: neuroblastoma cells=100∶1,50∶1,20∶1 respectively) on neuroblastoma cells was significantly higher than that in control group[(31.00 ±4.41)%,(30.92±5.27)%,(33.57±5.35)%,(26.23±5.20)%, t=3.51,2.98,4.24, P < 0.01); But the lethal effect of dendritic cells on neuroblastoma was significantly lower when their ratio was 100∶1 and 50∶1 in comparison with 20:1 (t=2.01,2.36, P < 0.05), and no significant difference in lethal effect existed between the ratio at 100∶1 and 50∶1(t=0.06,P > 0.05). CONCLUSION: Dendritic cells differentiated from human umbilical cord blood mononuclear cells after in vitro induction of cytokines can promote the lethal effect of T cells on neuroblastoma cells. The lethal effect is associated with the concentration of dendritic cells within some range.展开更多
文摘BACKGROUND: Cryoanalgesia at -50 ℃ for 90 seconds yields effective pain relief followingthoracotomy. In China, -50 ℃ is a common temperature for intercostal cryoanalgesia followingthoracotomy. However, experimental results vary.OBJECTIVE: To explore intercostal nerve pathological changes at-70 ℃ for various freezing timesby studying canines, and to evaluate long-term clinical efficacy of intercostal nerve cryoanalgesia forpostoperative pain relief based on the animal experiments.DESIGN, TIME AND SETTING: A comparative animal study was performed at the AnimalExperimental Center of the General Hospital of the People's Liberation Army. Based on results fromthe animal study, a randomized, controlled, clinical trial was performed at the Department ofThoracic Surgery of the General Hospital of the People's Liberation Army between October 2006and October 2008.PARTICIPANTS: A total of 120 patients undergoing posterolateral single incision Iobectomy at theDepartment of Thoracic Surgery of PLA General Hospital between October 2006 and October 2008were selected. Nervous system diseases were excluded.METHODS: Animal experiment: 8 anaesthetized, mixed-breed dogs were used. The intercostalnerves (costal bone 6-10) were frozen at -70 ℃ for varying times (30, 60, 90, 120, and 180seconds). Clinical study: 120 patients were randomly assigned to 2 groups (n = 60). In thecryoanalgesia group, the intercostal nerves were frozen prior to chest closure, and 4 costal nerves(1 at incision level, 2 above and below incision, and 1 at drainage tube level) were frozen for 90seconds at -70 ℃, respectively. Intercostal nerves were not frozen in the control group patients.Dolantin was used to relieve postoperative pain in patients from both groups.MAIN OUTCOME MEASURES: Pathological changes in frozen intercostal nerves were examined atdays 1, 10, 30, and 60 following freezing. Following surgery, the degree of postoperative pain in allpatients was evaluated by visual analogue scale at days 1,3, 5, 9, 30, 60, 90, and 180. Dolantindoses at days 1, 3, 5, 9 post-surgery and postoperative complications were noted.RESULTS: Nerve damage progressively increased with length of freezing time at-70 ℃, andrecovery time from damage was gradually increased. After freezing for 90 seconds, the nervesexhibited obvious histopathological damage, and then completely recovered. In addition, afterfreezing for 180 seconds, the histopathological changes in nerves were reversible. In the clinicalstudy, visual analogue scale scores were significantly less in the cryoanalgesia group compared withthe control group (P < 0.01), which was maintained over 30 days. In the cryoanalgesia group, themean dolantin dose administered and postoperative complications were significantly reducedcompared with the control group (P< 0.01).CONCLUSION: Freezing of the intercostal nerve at -70 ℃ for 90 seconds is a safe and long-termeffective method for relieving post-thoracotomy pain.
文摘BACKGROUND: Dendritic cell is the most major antigen presenting cell of organism. It is proved in recent studies that human umbilical cord blood mononuclear cells induced and cultured in vitro by recombinant human granulocyte-macrophage colony stimulating factor (rhG-MCSF) and recombinant human interleukin-4(rhIL-4) can generate a great many dendritic cells and promote the lethal effect of T cells on human neuroblastoma, but it is unclear that whether the lethal effect is associated with the most proper concentration of dendritic cells. OBJECTIVE: To investigate the lethal effect of human umbilical cord blood mononuclear cells induced in vitro by cytokines differentiating into dendritic cells on human neuroblastoma, and its best concentration range. DESIGN: Open experiment. SETTING: Department of Pediatrics, the Medical School Hospital of Qingdao University. MATERIALS: The study was carried out in the Shandong Provincial Key Laboratory (Laboratory for the Department of Pediatrics of the Medical School Hospital of Qingdao University) during September 2005 to May 2006. Human umbilical cord blood samples were taken from the healthy newborn infants of full-term normal delivery during October to November 2005 in the Medical School Hospital of Qingdao University, and were voluntarily donated by the puerperas. Main instruments: type 3111 CO2 incubator (Forma Scientific, USA), type 550 ELISA Reader (Bio-Rad, USA). Main reagents: neuroblastoma cell line SK-N-SH (Shanghai Institute of Life Science, Chinese Academy of Sciences), RPMI-1640 culture fluid and fetal bovine serum (Hyclone), rhIL-4 (Promega, USA), rhG-MCSF (Harbin Pharmaceutic Group Bioengineering Co.Ltd), rat anti-human CD1a monoclonal antibody and FITC-labeled rabbit anti-rat IgG (Xiehe Stem cell Gene Engineering Co.Ltd). METHODS: ① Human umbilical cord blood mononuclear cells obtained with attachment methods differentiated into human umbilical cord blood dendritic cells, presenting typical morphology of dendritic cells after in vitro induction by rhG-MCSF and rhIL-4. ② Different concentrations of dendritic cells[ dendritic cells: neuroblastoma cells=20∶1,50∶1,100∶1(2×108 L-1,5×108 L-1,1×109 L-1)], 1×109 L-1 T cells and 1×107 L-1 neuroblastoma cells were added in the experimental group. 1×109 L-1 T cells and 1×107 L-1 neuroblastoma cells were added in the control group. ③ Main surface marker CD1a molecules of dendritic cells were detected with indirect immunofluorescence, and the percent rate of dendritic cells was counted with ultraviolet light and expressed as the expression rate of CD1a+ cells. ④ Single effector cells and target cells were respectively set in the experimental group and control group to obtain the lethal effect. The lethal effect of dendritic cells on neuroblastoma cells was indirectly evaluated by detecting cellular survival with MTT assay. The lethal effect(%)=(1-A experimental well-A effector cell well/A target cell well)×100%.⑤The experimental data were presented as Mean ±SD, and paired t test was used. MAIN OUTCOME MEASURES: ① Morphological characters of dendritic cells in the process of induction and differentiation. ②CD1a+ cellular expression rate. ③Lethal effect of dendritic cells on neuroblastoma cells. RESULTS: ①Morphological characters of dendritic cells in the process of induction and differentiation: On the 15th day after human umbilical cord blood mononuclear cells were induced by rhG-MCSF and rhIL-4, typical morphology of dendritic cells could be seen under an inverted microscope. ②Expression rate of CD1a+ cells was (43.12±5.83)%. ③Lethal effect of dendritic cells on neuroblastoma cells: Lethal effect of dendritic cells stimulated T cells in each experimental group ( dendritic cells: neuroblastoma cells=100∶1,50∶1,20∶1 respectively) on neuroblastoma cells was significantly higher than that in control group[(31.00 ±4.41)%,(30.92±5.27)%,(33.57±5.35)%,(26.23±5.20)%, t=3.51,2.98,4.24, P < 0.01); But the lethal effect of dendritic cells on neuroblastoma was significantly lower when their ratio was 100∶1 and 50∶1 in comparison with 20:1 (t=2.01,2.36, P < 0.05), and no significant difference in lethal effect existed between the ratio at 100∶1 and 50∶1(t=0.06,P > 0.05). CONCLUSION: Dendritic cells differentiated from human umbilical cord blood mononuclear cells after in vitro induction of cytokines can promote the lethal effect of T cells on neuroblastoma cells. The lethal effect is associated with the concentration of dendritic cells within some range.
