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Objective@#To investigate the role and mechanism of Ly6Chigh monocyte in mice with ventilator-induced lung injury (VILI).@*Methods@#Forty-eight healthy male SPF C57BL/6 mice were divided into spontaneous breathing group (n = 8), normal tidal volume (VT) group (VT was 8 mL/kg, n = 8), and high VT group (VT was 20 mL/kg, n = 32). The mice in the high VT group were subdivided into 1, 2, 3 and 4 hours subgroups, with 8 mice in each subgroup. All mice underwent direct tracheal intubation, those in the spontaneous breathing group maintained spontaneous breathing, and those in the normal VT group and high VT group were mechanically ventilated with different VT. After ventilation for 4 hours, bronchoalveolar lavage fluid (BALF) was collected to determine total protein, and the levels of inflammatory factors including tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were determined by enzyme-linked immune sorbent assay (ELISA). The lung tissues were harvested to determine the wet/dry (W/D) ratio, and lung tissue injury was assessed in terms of lung histopathologic examination after hematoxylin-eosin (HE) staining under the light microscope. The protein expressions of monocyte chemotactic protein-1 (MCP-1) and CC-chemokine receptor 2 (CCR2) in lung tissues were determined by Western Blot. Flow cytometry was used to detect the proportion of Ly6Chigh monocyte in lung tissue.@*Results@#The histopathology of lung tissue structures was normal in the spontaneous breathing group and the normal VT group. Inflammatory reaction began to appear at 2 hours of high VT ventilation, and inflammatory reaction was gradually aggravated with the time extension. Compared with the spontaneous breathing group, the total protein, TNF-α, and IL-1β levels in BALF, the lung W/D ratio and MCP-1 expression were increased from 2 hours of high VT ventilation [total protein in BALF (g/L): 1.05±0.13 vs. 0.58±0.11, TNF-α in BALF (ng/L): 116.86±16.14 vs. 38.27±8.00, IL-1β in BALF (ng/L): 178.98±10.41 vs. 117.56±23.40, lung W/D ratio: 5.76±0.27 vs. 4.98±0.39, MCP-1/GAPDH: 0.87±0.19 vs. 0.29±0.12, all P < 0.05], and CCR2 expression and the proportion of Ly6Chigh monocyte was significantly increased from 3 hours of high VT ventilation [CCR2/GAPDH: 0.84±0.19 vs. 0.24±0.11, Ly6Chigh monocyte proportion: (9.01±2.47)% vs. (1.06±0.35)%, both P < 0.05], and they all showed an increased tendency with the time extension. There was no significant difference in the parameters mentioned above among the spontaneous breathing group, normal VT group and high VT ventilation 1-hour group.@*Conclusion@#Ly6Chigh monocytes are involved in VILI, which aggravate VILI by activating the MCP-1/CCR2 axis.
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Objective To investigate the role and mechanism of Ly6Chigh monocyte in mice with ventilator-induced lung injury (VILI). Methods Forty-eight healthy male SPF C57BL/6 mice were divided into spontaneous breathing group (n = 8), normal tidal volume (VT) group (VT was 8 mL/kg, n = 8), and high VT group (VT was 20 mL/kg, n = 32). The mice in the high VT group were subdivided into 1, 2, 3 and 4 hours subgroups, with 8 mice in each subgroup. All mice underwent direct tracheal intubation, those in the spontaneous breathing group maintained spontaneous breathing, and those in the normal VT group and high VT group were mechanically ventilated with different VT. After ventilation for 4 hours, bronchoalveolar lavage fluid (BALF) was collected to determine total protein, and the levels of inflammatory factors including tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were determined by enzyme-linked immune sorbent assay (ELISA). The lung tissues were harvested to determine the wet/dry (W/D) ratio, and lung tissue injury was assessed in terms of lung histopathologic examination after hematoxylin-eosin (HE) staining under the light microscope. The protein expressions of monocyte chemotactic protein-1 (MCP-1) and CC-chemokine receptor 2 (CCR2) in lung tissues were determined by Western Blot. Flow cytometry was used to detect the proportion of Ly6Chigh monocyte in lung tissue. Results The histopathology of lung tissue structures was normal in the spontaneous breathing group and the normal VT group. Inflammatory reaction began to appear at 2 hours of high VT ventilation, and inflammatory reaction was gradually aggravated with the time extension. Compared with the spontaneous breathing group, the total protein, TNF-α, and IL-1β levels in BALF, the lung W/D ratio and MCP-1 expression were increased from 2 hours of high VT ventilation [total protein in BALF (g/L): 1.05±0.13 vs. 0.58±0.11, TNF-α in BALF (ng/L): 116.86±16.14 vs. 38.27±8.00, IL-1β in BALF (ng/L): 178.98±10.41 vs. 117.56±23.40, lung W/D ratio: 5.76±0.27 vs. 4.98±0.39, MCP-1/GAPDH: 0.87±0.19 vs. 0.29±0.12, all P < 0.05], and CCR2 expression and the proportion of Ly6Chigh monocyte was significantly increased from 3 hours of high VT ventilation [CCR2/GAPDH:0.84±0.19 vs. 0.24±0.11, Ly6Chigh monocyte proportion: (9.01±2.47)% vs. (1.06±0.35)%, both P < 0.05], and they all showed an increased tendency with the time extension. There was no significant difference in the parameters mentioned above among the spontaneous breathing group, normal VT group and high VT ventilation 1-hour group. Conclusion Ly6Chigh monocytes are involved in VILI, which aggravate VILI by activating the MCP-1/CCR2 axis.
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The two-dimensional (2D) cell culture model is currently used to study cellular processes and drug screening for human diseases. However, the growth of cells is affected by many factors. For conventional 2D cell culture, many of the difficulties are encountered in accurately replicating the cell function in three-dimensional (3D) tissues. Compared with 2D cell culture, much attention is paid to the cell-to-cell and cell-matrix interactions for 3D cell culture systems, which can more closely mimic the growth environment for cultured cells. Therefore, the 3D cell culture system was more suitable for a variety of applications such as drug screening and cell proliferation. In this work, we prepared microarray-structured polymer films with different geometric structures by nanoimprint lithography and used the films as cell culture platforms for the culture of 293T cells. Through the adjustment of the surface morphology and water contact angle of the prepared films, the regulation of the morphological changes of cell growth was successfully realized. Experimental results demonstrated that the hydrophilic films with 10 μm-pillar microstructure are applicable to 3D cell culture, whereas the hydrophobic films with 3 μm-pillar microstructure are only suitable for 3D culture of cells with a smaller size and stiff cuticular layer. In addition, cells tended to the formation of spheroids on the hydrophobic films, while cells usually adhered to the surface and grew on the hydrophilic films. This work represents further technological progress in the development of 3D cell culture, thereby facilitating future studies of physiologically relevant processes.
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Humans , Cell Culture Techniques , Cell Proliferation , HEK293 Cells , PolymersABSTRACT
Objective To conclude the diagnostic and therapeutic experience for acute mesenter ic ischemia(AMI).Methods Clinical data of 102 AMI patients were analyzed retrospectively.The curative effect of conservative management,endovascular treatment and open surgery were evaluated.All patients were treated with anticoagulation,vasodilator,thrombolysis after the diagnosis was made.17 cases were relieved by simple medical therapy.The other 85 patients had no obvious effect within 48 hours after conservative treatment.Of the 63 cases undergone endovascular treatment,58 were relieved.Among other 7 unrelieved cases,2 of them gave up treatment and 5 patients were turned to open surgery.The left 22 cases were converted to open surgery directly.Results Of all 102 cases,92 patients were relieved completely,5 patients were turned to chronic intestinal dysfunction.3 patients died in hospital and 2 patients lost follow-up.Conclusion AMI is a potentially fatal vascular emergency.Early diagnosis by CTA and revascularization of intestine without delay is the pivotal management.Endovascular treatment is the preferred therapeutic options.
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Objective To explore the efficacy and safety of microwave hemostatic device in laparoscopic microwave ablation for exogenic hepatic hemagioma.Methods The clinic data of 62 patients with exogenic hepatic hemagioma who performed with laparoscopic microwave ablation between May 2015 and May 2017 were retrospectively analyzed.According to the different surgical technique,the patients were assigned into microwave hemostatic device combined with microwave ablation group (combination group,29 patients) and microwave ablation group (microwave group,33 patients).The microwave group was performed laparoscopic microwave ablation,and the combination group was pretreated the surface of hemagioma with microwave hemostatic device before laparoscopic microwave ablation.The intraoperative and postoperative conditions of the patients were recorded and analyzed.Results All patients were performed successfully under laparoscope without conversion to laparotomy.The average time of microwave ablation was significantly shorter in combination group than in microwave group [(10.69 ±3.54) min vs.(13.18 ± 4.31) min,P＜0.05].Compared with microwave group,the average bleeding amount of operation was significantly lower in combination group[(48.79±20.30) mL vs.(95.76±90.16) mL,P＜0.05].All patients from both groups recovered uneventfully without any complications such as abdominal bleeding or bile leakage.Conclusion For exogenic hepatic hemagioma,the microwave hemostatic device is used to solidify the surface of hemagioma before microwave ablation,which can improve the safety of the operation,reduce the time of microwave ablation,and avoid tumor hemorrhage caused by puncture.
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Objective To conclude the diagnostic and therapeutic experience for acute mesenter ic ischemia(AMI).Methods Clinical data of 102 AMI patients were analyzed retrospectively.The curative effect of conservative management,endovascular treatment and open surgery were evaluated.All patients were treated with anticoagulation,vasodilator,thrombolysis after the diagnosis was made.17 cases were relieved by simple medical therapy.The other 85 patients had no obvious effect within 48 hours after conservative treatment.Of the 63 cases undergone endovascular treatment,58 were relieved.Among other 7 unrelieved cases,2 of them gave up treatment and 5 patients were turned to open surgery.The left 22 cases were converted to open surgery directly.Results Of all 102 cases,92 patients were relieved completely,5 patients were turned to chronic intestinal dysfunction.3 patients died in hospital and 2 patients lost follow-up.Conclusion AMI is a potentially fatal vascular emergency.Early diagnosis by CTA and revascularization of intestine without delay is the pivotal management.Endovascular treatment is the preferred therapeutic options.
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Objective To explore the efficacy and safety of microwave hemostatic device in laparoscopic microwave ablation for exogenic hepatic hemagioma.Methods The clinic data of 62 patients with exogenic hepatic hemagioma who performed with laparoscopic microwave ablation between May 2015 and May 2017 were retrospectively analyzed.According to the different surgical technique,the patients were assigned into microwave hemostatic device combined with microwave ablation group (combination group,29 patients) and microwave ablation group (microwave group,33 patients).The microwave group was performed laparoscopic microwave ablation,and the combination group was pretreated the surface of hemagioma with microwave hemostatic device before laparoscopic microwave ablation.The intraoperative and postoperative conditions of the patients were recorded and analyzed.Results All patients were performed successfully under laparoscope without conversion to laparotomy.The average time of microwave ablation was significantly shorter in combination group than in microwave group [(10.69 ±3.54) min vs.(13.18 ± 4.31) min,P＜0.05].Compared with microwave group,the average bleeding amount of operation was significantly lower in combination group[(48.79±20.30) mL vs.(95.76±90.16) mL,P＜0.05].All patients from both groups recovered uneventfully without any complications such as abdominal bleeding or bile leakage.Conclusion For exogenic hepatic hemagioma,the microwave hemostatic device is used to solidify the surface of hemagioma before microwave ablation,which can improve the safety of the operation,reduce the time of microwave ablation,and avoid tumor hemorrhage caused by puncture.
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ObjectiveTo investigate the role and its mechanism of the NOD-like receptor 3 (NLRP3) inflammasome in alveolar macrophages in ventilator-induced lung injury (VILI) in rats.Methods Thirty adult male Sprague-Dawley (SD) rats were randomly divided into three groups, with 10 rats in each group: spontaneous breathing control group, normal tidal volume (VT) group (VT 8 mL/kg) and high VT group (VT 40 mL/kg). All of the rats underwent tracheotomy. Then rats in spontaneous breathing control group were kept to have spontaneous breathing, while rats in normal VT group and high VT group received mechanical ventilation. After 4 hours, the rats were sacrificed by carotid artery bleeding, and the bronchoalveolar lavage fluid (BALF), blood serum and lung tissue were collected. Lung wet/dry ratios (W/D) were measured. Light microscopy and electron microscopy were performed to observe the pathological changes in lung tissue, and the ultrastructural changes in alveolar macrophages. Enzyme linked immunosorbent assay (ELISA) was performed to measure the total protein content in the BALF and the interleukins (IL-1β and IL-18) in the serum and BALF. The mRNA expressions and protein levels of NLRP3, apoptosis-associated speck-like protein containing CARD (ASC), caspase-1, and nuclear factor-κB (NF-κB) in alveolar macrophages were assayed by real-time fluorescent quantization reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot.Results The structure of lung tissue and alveolar macrophages of rats in spontaneous breathing control group and normal VT group appeared normal, while obvious inflammatory changes were found in high VT group. Compared with spontaneous breathing control group and normal VT group, the ratio of W/D (8.89±0.90 vs. 5.18±0.86, 5.71±0.82, bothP< 0.05), contents of total protein, IL-1β, IL-18 in BALF were significantly increased [total protein (g/L):2.34±0.41 vs. 1.77±0.14, 1.81±0.06, IL-1β (ng/L): 133.48±10.48 vs. 81.54±3.12, 83.80±5.22, IL-18 (μg/L):4.57±0.45 vs. 3.04±0.51, 3.43±0.43, allP< 0.05], and IL-1β and IL-18 in serum were also increased [IL-1β(ng/L): 105.06±10.18 vs. 65.11±8.58, 75.30±10.62, IL-18 (μg/L): 2.27±0.09 vs. 1.18±0.34, 1.43±0.15, all P< 0.05]. The mRNA and protein expressions of NLRP3, ASC, caspase-1 and NF-κB in alveolar macrophages of high VT group were significantly increased compared with that of spontaneous breathing control group and normal VT group. The mRNA expressions of NLRP3, ASC, caspase-1 and NF-κB in high VT group were (8.53±2.21), (5.75±1.17), (7.47±1.23) and (10.86±2.38) folds of those in spontaneous breathing control group, and the protein expressions of NLRP3, ASC, caspase-1 and NF-κB were (1.50±0.14), (1.49±0.04), (1.53±0.15) and (1.51±0.11) folds of those in spontaneous breathing control group (allP< 0.01). There were no significant differences in all the indexes between normal VT group and spontaneous breathing control group.ConclusionNLRP3 inflammasome in alveolar macrophages may be involved in the mechanism of occurrence of VILI.
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Objective To investigate the analgesic effects of parecoxib sodium combined with patient controlled epidural analgesia (PCEA) after orthopedic subarachnoid block anesthesia surgery.Methods Two hundred patients undergone orthopedic subarachnoid block anesthesia surgery were randomly and equally divided into two groups:Group P (treated intravenously with 40 mg parecoxib sodium combined with PCEA at the end of operation) and Group C ( treated intravenously with 0.5 g tramadol combined with PCEA at the end of operation).The visual analog scale (VAS) was performed at 6,12,24,48 and 72 hours postoperatively in two groups.Meanwhile,the press frequency of analgesic pump,effective frequency,side effects and satisfaction degree were recorded. Results The VAS sore of Group P was lower than that of Group C at 6,12,24,48 and 72 hours postoperatively ( P ＜ 0.05 ).Group P showed a less number in aspects of the press frequency of analgesic pump,effective frequency,and side effects at 12 and 24 hours,but a higher satisfactory degree,compared with Group C (P ＜0.05). Conclusion Combined use of parecoxib sodium and PCEA can exert a better analgesic effect and have a low incidence rate of side effects following orthopedic subarachnoid block anesthesia.
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In order to investigate the expression of Notch-1 in rats with acute pancreatitis (AP) and its relation with apoptosis of pancreatic cells, mild AP (MAP) and severe AP (SAP) models were established by retrograde injection of different concentrations of sodium taurocholae into pancreatic duct. The apoptosis index and the expression of Notch-1 protein and mRNA were detected by using TUNEL, Western blot and real-time PCR in MAP and SAP at different time intervals. The results showed that in MAP group the apoptosis index was significantly elevated during 4 to 24 h after induction of pancreatitis, but there was no significant difference among different time intervals. In SAP group, the apoptosis index reached the peak at 4th h after induction of pancreatitis, then gradually declined. There was significant difference in apoptosis index between MAP and SAP groups. Starting from 4th after induction of pancreatitis, the expression of Notch-1 in both MAP and SAP group was increased at different time intervals, but that in SAP group was significantly higher than in MAP group (P＜0.05). The expression of Notch in MAP and SAP groups reached the peak at 12th and 8th h respectively after induction of pancreatitis. It was concluded that there was significant difference in apoptosis index and the Notch-1 expression in different types of AP. The overexpression of Notch-1 could aggravate AP by inhibiting the apoptosis of pancreatic cells.
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In order to investigate the effects of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on the proliferation, apoptosis of pancreatic cancer cell line SW1990 cells and the ex-pression of cyclin E mRNA, the SW1990 cells were treated with different concentrations of EPA or DHA (20, 40, 60 μg/mL) for 0, 12, 24, 36 and 48 h respectively. By using MTT method, the inhibi-tory effects of EPA or DHA on the cell growth were assayed. Real time PCR was used to detect the expression changes of cyclin E mRNA after the SW1990 cells were treated with 40 μg/mL EPA or DHA for different time. Flow eytometry was used to test the changes of apoptostic rate in the SW1990 cells treated with different concentrations of EPA or DHA for 24 h. The results showed that EPA and DHA could inhibit the growth of SW1990 cells in a time- and concentration-dependent manner (P<0.01). EPA or DHA could also significantly inhibit the expression of cyclin E mRNA in a time-dependent manner (P<0.05). EPA or DHA could induce the apoptosis of SW1990 cells in a concentration-dependent manner (P<0.01). It was concluded that ω-3 fatty acid could inhibit the pro- liferation of pancreatic cancer cell line SW1990 cells and promote their apoptosis. The down-regulation of the cyclin E expression by ω-3 fatty acid might be one of the mechanisms for its anti-tumor effect on pancreatic cancer.
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In order to investigate the expression of Notch-1 in rats with acute pancreatitis (AP) and its relation with apoptosis of pancreatic cells, mild AP (MAP) and severe AP (SAP) models were established by retrograde injection of different concentrations of sodium taurocholae into pancreatic duct. The apoptosis index and the expression of Notch-1 protein and mRNA were detected by using TUNEL, Western blot and real-time PCR in MAP and SAP at different time intervals. The results showed that in MAP group the apoptosis index was significantly elevated during 4 to 24 h after induction of pancreatitis, but there was no significant difference among different time intervals. In SAP group, the apoptosis index reached the peak at 4th h after induction of pancreatitis, then gradually declined. There was significant difference in apoptosis index between MAP and SAP groups. Starting from 4th after induction of pancreatitis, the expression of Notch-1 in both MAP and SAP group was increased at different time intervals, but that in SAP group was significantly higher than in MAP group (P<0.05). The expression of Notch in MAP and SAP groups reached the peak at 12th and 8th h respectively after induction of pancreatitis. It was concluded that there was significant difference in apoptosis index and the Notch-1 expression in different types of AP. The overexpression of Notch-1 could aggravate AP by inhibiting the apoptosis of pancreatic cells.