1.Management of cardiopulmonary bypass with hypothermic circulatory arrest during aortic arch surgery
Weijun WANG ; Danfeng KANG ; Yunxia GE ; Yuan FENG ; Feng LIAN ; Genxing XU ; Song XUE
Clinical Medicine of China 2011;27(12):1253-1256
Objective To summarize our experience in the management of cardiopulmonary bypass (CPB) with deep hypothermic circulatory arrest (DHCA) during aortic arch surgery.Methods From March 2007 to May 2010,58 consecutive patients,including 24 urgent and 34 selective operations underwent aortic arch surgery.Thirty-nine hemiarch and 19 total aortic arch replacement operations were performed.CPB was established by perfusion through femoral artery (42 cases) and right subclavian artery (RSA) ( 16 cases),of which 4 cases were carried out with antegrade cerebral perfusion (ACP).Results The mean CPB time was ( 208.88 ± 136.45 ) min.The mean cerebral circulation arrest was ( 27.36 ± 11.50 ) min.Nasopharyngeal and rectal temperature were ( 16.01 ±2.67)℃ and ( 19.72 ±2.13)℃ respectively before DHCA was initiated.The mean times for cooling and rewarming were ( 50.91 ± 16.89) min and ( 88.97 ± 43.68 ) min.The mean time of intubation was (56.70 ± 45.19 ) h.The time in ICU was ( 5.68 ± 5.31 ) d,and the time of hospitalization was (30.11 ± 22.27 ) d.Acute renal failure,hypoxemia,and paraplegia occurred post-operatively in 4,19,and 2 patients,respectively.Four patients died post-operatively with a mortality of 6.90%.Compared with those received hemiarch replacement operation,the patients received total aortic arch replacement had statistically longer time of CPB([262.16 ±219.97]min vs [182.92 ±53.81] min,t =2.14,P <0.05),cerebral circulatory arrest ( [30.47 ± 15.86 ] win vs [25.85 ± 8.48 ] min,t =2.40,P < 0.05 ),rewarming ( [110.00 ± 68.66 ] min vs [78.72 ± 17.31 ] min,t =2.69,P < 0.05 ),and intubation ( [93.95 ± 131.89 ] h vs [38.08 ± 30.70 ] h,t =2.50,P < 0.05 ).There was no significant difference in the times of these procedures between emergency surgery group and elective surgery group,between RSA and femoral artery cannulation groups.Conclusion It is crucial that the cooling and re-warming procedures during aortic arch surgery should be carried out slowly,gradually,and completely when DHCA was adopted alone.conclusion through right axillary artery or RSA was preferred for ACP,in order to accomplish the body circulation arrest at a relative high temperature,to shorten the CPB time,and to alleviate potential harmful effects of hypothermia.Meticulous management of CPB is one of the most important measures to improve the patients' outcome.
2.Establishment of small intestinal organoid models in a novel culture system
Bian WU ; Guili LIANG ; Chengfeng XING ; Zaozao WU ; Junmo WU ; Yu KANG ; Yinglei MIAO ; Danfeng LAN
Chinese Journal of Digestion 2023;43(11):764-770
Objective:To establish a new type of small intestinal organoids with injury-related regenerative capacity, and to simulate the process of intestinal injury, regeneration, and repair in vitro. Methods:The crypt structures of ileal mucosa from 6 to 8 weeks old, 18 to 24 g specific pathogen-free C57BL/6 mice were isolated. The ENR, ENR+ tumor necrosis factor-α(TNF-α) and 8C culture systems were designed to establish small intestinal organoids under conditions of intestinal homeostasis, inflammatory injury and injury-related regeneration, and the morphology of intestinal organoids were observed. The cell types and spatial arrangements of intestinal organoids, and the expression of genes clusterin( Clu), annexin A1( Anxa1), stem cell antigen-1( Sca1) and regenerating islet-derived protein 3-beta( Reg3 b) at protein levels were detected by immunofluorescence staining. The expression of genes Clu, Anxa1, Sca1 and Reg3 b at mRNA levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Independent sample- t test was used for statistical analysis. Results:In ENR and 8C culture system, both intestinal organoids contained intestinal stem cells, goblet cells, Paneth cells and intestinal endocrine cells, and the spatial arrangement of cells was similar to the intestinal epithelium. In the 8C culture system, the amplification capacity of the new small intestinal organoids was significantly enhanced, the growth rate was faster, and the structure was larger and more complex than those of small intestinal organoids in ENR and ENR+ TNF-α culture systems. The results of qRT-PCR showed that, the relative mRNA expression levels of novel small intestinal organoid regeneration genes Clu, Anxa1, and Sca1 in the 8C culture system were higher than those in the ENR and ENR+ TNF-α culture systems (0.68±0.31 vs.0.20±0.07 and 0.36±0.19, 0.48±0.13 vs. 0.07±0.02 and 0.18±0.11, 0.56±0.20 vs. 0.02±0.01 and 0.08±0.04), and the differences were statistically significant ( t=4.82 and 2.77, 8.62 and 4.89, and 8.58 and 7.50; all P<0.05). The results of immunofluorescence staining indicated that, the expression levels of novel small intestinal organoid regeneration genes Clu, Anxa1, Sca1 and Reg3 b at protein level in the 8C culture system were higher than those in the ENR and ENR+ TNF-α culture systems (31.62±1.69 vs. 9.73±2.39 and 15.11±2.16, 42.65±1.85 vs. 19.70±1.18 and 24.97±2.82, 63.80±2.73 vs. 37.10±1.59 and 43.27±2.53, 53.26±1.84 vs. 27.75±3.78 and 33.16±3.50), and the differences were statistically significant( t=12.95 and 10.41, 18.13 and 9.09, 14.63 and 9.56, and 10.51 and 8.80; all P<0.001). Conclusion:The small intestinal organoids established in the novel culture system have the characteristics of injury-related regeneration, and provide a novel in vitro model for studying the regeneration of epithelial tissues and organs.