目的 建立兔腰椎间盘严重退变骨水泥成形术模型并进行鉴定。方法 选用新西兰白兔6只,手术干预前摄腰椎正侧位X线片并进行MRI扫描Pfirrmann分级,之后通过腹外斜肌与腰大肌间隙入路手术去除兔腰2~3椎间盘髓核组织及部分纤维环模拟腰椎间盘严重退变状态。饲养6周后相应腰椎节段椎间盘进行MR扫描Pfirrmann分级,确认相应腰椎节段椎间盘符合严重退变影像表现后再次手术显露相应椎间隙并注入骨水泥。1周后再次摄腰椎正侧位X线片并行MRI扫描Pfirrmann分级,终末处死并解剖动物检查椎间盘内骨水泥填充情况。结果 兔腰椎间盘退化模型建立6周后磁共振Pfirrmann分级为Ⅴ级。椎间隙骨水泥注射后1周其术后磁共振Pfirrmann分级为Ⅳ。骨水泥注射模型1周后拍摄手术节段X线片显示骨水泥较好地填充于腰2~3间隙,椎间隙高度接近正常状态。终末处死并解剖动物发现腰椎节段椎间盘内骨水泥填充良好无脱落或松动。结论 通过腹外斜肌与腰大肌间隙入路,手术去除椎间盘髓核组织及部分纤维环6周后,往椎间隙内注入骨水泥,可获得较为可靠的新西兰大白兔腰椎间盘严重退变骨水泥成形术的动物模型。

Objective To establish and identify the rabbit model of lumbar disc with severe degeneration．Methods Six New Zealand white rabbits were selected,lumbar X-ray and Pfirrmann grade by MR scan were performed before surgical intervention．Along the space of obliquus externus abdominis and psoas major,the front edge of L2 to L3 was exposed．Then,the nucleus pulposus and part of annulus fibrosus were removed to imitate severe degeneration of lumbar disc．After 6 weeks of rearing,the operated lumbar disc was graded by MR scan,confirming that the lumbar disc met the image of severe degeneration,and then exposed the intervertebral space and injected bone cement．One week later,the anterior lumbar X-ray and the MRI scan for Pfirrmann grading were taken．The animals were sacrificed and dissected to check the bone cement filling in the intervertebral disc．Results The rabbit MR Pfirrmann grade of intervertebral disk was V after 6 weeks of first operation．One week after intervertebral cement injection,the MR Pfirrmann grade was Ⅳ．The surgical segment X-ray was taken one week after the cement injection,which showed that the cement was well filled in the L2-L3 gap and the vertebral space height was close to normal．Animals were sacrificed and dissected,the lumbar intervertebral disc was well filled with cement without shedding or loosening．Conclusions A reliable animal model of lumbar disc with severe degeneration in New Zealand white rabbits can be obtained by injecting cement into the intervertebral space after 6 weeks of removal of the intervertebral disc nucleus pulposus and part of the annulus fibrosus through the obliquus externus abdomins and psoas major intervertebral space．

目的 探讨新生儿坏死性小肠结肠炎(NEC)炎症损伤与肠道微生态-LPS-TLR4通路之间的关系。方法 本研究收集2019年3月1日—2021年1月31日在中山市人民医院新生儿监护室确诊为NEC新生儿11例为实验组,随机选取30 例同期在新生儿科病房住院喂养顺利,排除NEC及败血症诊断的新生儿为对照组。采集2组新生儿的粪便标本,进行Real-time PCR表达谱分析2组粪便肠道菌群;取2组外周静脉血检测外周血单核细胞Toll样受体4(TLR4)和血清PCT、CRP、IL-6、SAA等指标,对比2组肠道菌群、外周血单核细胞TLR4和炎症指标水平,通过统计学分析组间差异。结果 本研究结果提示实验组变形菌门占82%(9/11),厚壁菌门占9%(1/11),放线菌门占9%(1/11),对照组变形菌门占20%(6/30),厚壁菌门占73%(22/30),放线菌门占7%(2/30),2组患儿的粪便肠道菌群分布有差异(χ²=11.521,P<0.05);实验组患儿外周血单核细胞TLR4水平高于对照组,组间差异有统计学意义(P<0.001);实验组患儿血清PCT、CRP、IL-6和SAA等炎症指标高于对照组,组间差异有统计学意义(P<0.001)。结论 NEC患儿的肠道菌群以变形菌门为主,伴外周血单核细胞TLR4和外周血炎症指标升高。可见,肠道微生态-LPS-TLR4通路可能与新生儿坏死性小肠结肠炎炎症损伤相关,具体的机制仍需进一步深入研究。

Objective To investigate the relationship between intestinal flora-LPS-TLR4 pathway and the inflammatory injury of neonatal necrotizing enterocolitis (NEC). Methods Eleven neonates with NEC from March, 2019 to January, 2021 were enrolled as the experimental group, and 30 neonates without NEC and septicemia who were admitted in the department of neonatology in the same period were included as the control group. Faecal flora from the two groups were collected and analyzed by Real-time PCR. Toll-like receptor 4 (TLR4) and serum PCT, CRP, IL-6, SAA in peripheral blood were measured. The intestinal flora, the expression of TLR4 in peripheral blood leukocytes and inflammatory markers were compared between two groups. Results It showed that the ratio of Proteobacteria was 82% (9/11), Firmicutes was 9% (1/11), Actinobacteria was 9% (1/11) in the experimental group. In the control group, the ratio of Proteobacteria was 20% (6/30), Firmicutes was 73% (22/30), Actinobacteria was 7% (2/30). There was a significant difference in the distribution of faecal flora between the two groups (χ² = 11.521, P<0.05), and the level of TLR4 in peripheral blood of the experimental group was significantly higher than that of the control group (P＜0.001). The levels of serum PCT, CRP, IL-6 and SAA in the experimental group were significantly higher than those in the control group (P＜0.001). Conclusions The main intestinal flora of neonates with NEC is Proteobacteria, with elevated TLR4 expression and inflammatory markers in peripheral blood. Therefore, the intestinal flora-LPS-TLR4 pathway may be associated with inflammatory injury in neonatal necrotizing enterocolitis.The specific mechanism still needs further study.