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目的探讨人离体子宫组氨酸-色氨酸-酮戊二酸盐(HTK)液冷灌注及冷缺血保存的安全区间。方法取因宫颈癌行广泛子宫切除术的患者子宫7例,3例于低温条件下用HTK液经子宫动脉灌注,测定一定灌注压力所对应的灌注高度,建立灌注高度与灌注压之间关系的数学模型;4例在适当灌注高度下短时灌注后取子宫内膜及肌组织于HTK保存液和生理盐水中保存,两组又根据保存时间(0、3、6、24 h)的不同分为0 h组、HTK 3 h组、HTK 6 h组、HTK 24 h组、生理盐水3 h组、生理盐水6 h组和生理盐水24 h组,测定不同保存时限的子宫组织细胞形态及平滑肌收缩能力,分析离体子宫灌注后冷缺血最佳保存时限。结果灌注压90~140 mm Hg对应的灌注高度为62~122 cm;与0 h组比较,HTK3 h组和HTK6 h组的子宫标本在光镜及电镜下形态学结构均未见明显改变,但HTK 24 h组及所有的生理盐水保存组的样本均出现不同程度的细胞水肿、细胞间失去联系、线粒体肿胀及染色质粗染等不可逆的退化性改变;HTK 3 h、HTK 6 h和HTK 24 h组的3组标本的肌收缩力差异均无统计学意义(P=0.772);而与相同时段HTK组标本相比,生理盐水3 h、6 h和24 h组标本的肌收缩最大波幅均明显降低(P<0.05)。结论人类离体子宫组织低温HTK液安全灌注高度是62~122 cm,短时灌注后4℃HTK液保存状态下可耐受冷缺血至少6 h。
Objective To investigate the safety interval of cryogenic perfusion and cryogenic preservation of human uterine histidine-tryptophan-ketoglutarate (HTK) in vitro. Methods Seven patients with uterine cancer underwent extensive hysterectomy due to cervical cancer. Three patients underwent uterine artery perfusion with HTK solution at low temperature. The perfusion height corresponding to the perfusion pressure was measured to establish the relationship between perfusion height and perfusion pressure The model of endometrium and muscle tissue was preserved in HTK preservation solution and saline solution in 4 cases under the appropriate perfusion height. The two groups were preserved according to the different preservation time (0, 3, 6, 24 h) The cells in 0 h group, HTK 3 h group, HTK 6 h group, HTK 24 h group, physiological saline 3 h group, saline 6 h group and saline 24 h group were used to determine the morphology and smooth muscle contraction Ability to analyze the best preservation time of cold ischemia after uterine perfusion. Results The perfusion heights corresponding to perfusion pressure of 90-140 mm Hg ranged from 62 to 122 cm. Compared with 0 h group, the morphological changes of hysterical specimens in HTK3 h group and HTK6 h group showed no obvious changes under light and electron microscopy, HTK 24 h group and all the saline preservation group samples showed varying degrees of cell edema, cell loss, irreversible degenerative changes such as mitochondria swelling and chromatin staining; HTK 3 h, HTK 6 h and HTK 24 h group had no significant difference in muscle contractility (P = 0.772). Compared with HTK group at the same time point, the maximal amplitude of muscle contraction in 3 h, 6 h and 24 h groups Significantly lower (P <0.05). Conclusions The safe perfusion of hypothermic HTK solution in human uterine tissue is 62 ~ 122 cm. Cold ischemia can be tolerated for at least 6 h after storage at 4 ℃ for 4 h under short-term perfusion.