Umbilical cord mesenchymal stem cell exosomes alleviate necrotizing enterocolitis in neonatal mice by regulating intestinal epithelial cells autophagy

BACKGROUND Necrotizing enterocolitis(NEC)is a severe gastrointestinal disease that affects premature infants.Although mounting evidence supports the therapeutic effect of exosomes on NEC,the underlying mechanisms remain unclear.AIM To investigate the mechanisms underlying the regulation of inflammatory response and intestinal barrier function by umbilical cord mesenchymal stem cell(UCMSCs)exosomes,as well as their potential in alleviating NEC in neonatal mice.METHODS NEC was induced in 5-d-old C57BL/6 pups through hypoxia and gavage feeding of formula containing lipopolysaccharide(LPS),after which the mice received human UCMSC exosomes(hUCMSC-exos).The control mice were allowed to breastfeed with their dams.Ileal tissues were collected from the mice and analyzed by histopathology and immunoblotting.Colon tissues were collected from NEC neonates and analyzed by immunofluorescence.Molecular biology and cell culture approaches were employed to study the related mechanisms in intestinal epithelial cells.RESULTS We found that autophagy is overactivated in intestinal epithelial cells during NEC,resulting in reduced expression of tight junction proteins and an increased inflammatory response.The ability of hUCMSC-exos to ameliorate NEC in a mouse model was dependent on decreased intestinal autophagy.We also showed that hUCMSC-exos alleviate the inflammatory response and increase migration ability in intestinal epithelial cells induced by LPS.CONCLUSION These results contribute to a better understanding of the protective mechanisms of hUCMSC-exos against NEC and provide a new theoretical and experimental foundation for NEC treatment.These findings also enhance our understanding of the role of the autophagy mechanism in NEC,offering potential avenues for identifying new therapeutic targets.

Efficacy of peritoneal drainage in very-low-birth-weight neonates with Bell’s stage II necrotizing enterocolitis:A single-center retrospective study

BACKGROUND Currently,pediatric surgeons are challenged by a lack of consensus on the optimal management strategy(conservative or surgical)for children with Bell’s stage II necrotizing enterocolitis(NEC).AIM To evaluate the clinical efficacy of peritoneal drainage in very-low-birth-weight(VLBW)neonates with modified Bell’s stage II NEC.METHODS This was a retrospective analysis of 102 NEC(modified Bell’s stage II)neonates born with VLBW who were treated at the Fujian Children’s Hospital(Fujian Branch of Shanghai Children’s Medical Center)between January 2017 and January 2020;these included 24 cases in the peritoneal drainage group,36 cases in the exploratory laparotomy group,and 42 cases in the conservative treatment group.RESULTS The general characteristics were comparable in the three groups(P>0.05).Compared with conservative treatment,peritoneal drainage was associated with significantly shorter fasting time,abdominal distension relief time,fecal occult blood(OB)negative conversion time,and reduced hospital length of stay(HLOS)(P<0.05 for all).Despite some advantages of peritoneal drainage over conservative treatment in terms of cure,conversion to laparotomy,intestinal perforation,intestinal stenosis,and abdominal abscess rates,the differences were not statistically significant(P>0.05).Compared to exploratory laparotomy,the fecal OB negative conversion time was significantly shorter in the peritoneal drainage group(P<0.05);similarly,the exploratory laparotomy group showed longer fasting time,abdominal distension relief time,HLOS,and higher complication rate compared to peritoneal drainage group,but the between-group differences were not statistically significant(P>0.05).CONCLUSION Peritoneal drainage,an easy-to-operate procedure,can improve the clinical symptoms of VLBW neonates with Bell’s stage II NEC and help reduce the HLOS.

Role of LPS/CD14/TLR4-mediated inflammation in necrotizing enterocolitis:Pathogenesis and therapeutic implications

AIM:To establish the roles of lipopolysaccharide (LPS)/CD14/toll-like receptor 4 (TLR4)-mediated inflammation in a rat model of human necrotizing enterocolitis (NEC).METHODS: Six pairs of intestinal samples from human NEC were collected before and after recovery for histological and molecular analysis of inflammatory cytokines and signaling components. In the rat NEC model, we isolated 10-cm jejunum segments and divided them into six groups (n=6) for sham operation, treatment with LPS, bowel distension, combined bowel distension and LPS stimulation, and two therapeutic groups. The potential eff icacy of a recombinant CD18 peptide and a monoclonal CD14 antibody was evaluated in the latter two groups. The serum and tissue levels of several inflammatory mediators were quantified by real-time polymerase chain reaction, ELISA and immunoblotting.RESULTS: Human acute phase NEC tissues displayed significant increases (P<0.05) in levels of TLR4, CD14, myeloid differentiation protein (MD)-2, tumor necrosis factor (TNF)-α and nuclear factor-κB when compared to those after recovery. The histological and inflammatory picture of human NEC was reproduced in rats that were treated with combined bowel distension and LPS, but not in the sham-operated and other control rats. Serum levels of interleukin-6 and TNF-α were also elevated. The NEC pathology was attenuated by treating the NEC rats with a monoclonal CD14 antibody or an LPS-neutralizing peptide.CONCLUSION:LPS and distension are required to produce the histological and inflammatory features of NEC. A potential treatment option is blocking LPS activation and leukocyte infi ltration.

Necrotizing enterocolitis: A multifactorial disease with no cure

Necrotizing enterocolitis is an inflammatory bowel disease of neonates with significant morbidity and mortality in preterm infants. Due to the multifactorial nature o the disease and limitations in disease models, early diagnosis remains challenging and the pathogenesis elusive. Although preterm birth, hypoxic-ischemic events formula feeding, and abnormal bacteria colonization are established risk factors, the role of genetics and vasoactive/inflammatory mediators is unclear Consequently, treatments do not target the specific underlying disease processes and are symptomatic and surgically invasive. Breast-feeding is the most effective preventative measure. Recent advances in the prevention of necrotizing enterocolitis have focused on bioactive nutrients and trophic factors in human milk. Developmen of new disease models including the aspect of prematurity that consistently predisposes neonates to the disease with multiple risk factors will improve our understanding of the pathogenesis and lead to discovery of innovative therapeutics.

Prevention of necrotizing enterocolitis in premature infants——an updated review

Necrotizing enterocolitis(NEC) is among the most common and devastating diseases encountered in premature infants, yet the true etiology continues to be poorly understood despite decades of research. Recently, gut bacterial dysbiosis has been proposed as a risk factor for the development of NEC. Based on this theory, several best clinical practices designed to reduce the risk of NEC have been proposed and/or implemented. This review summarizes the results of recent clinical trials and meta-analyses that support some of the existing clinical practices for reducing the risk of NEC in premature infants. It is evident that human milk feeding can reduce the incidence of NEC. While most of the studies demonstrated that probiotic supplementation can significantly reduce the incidence of NEC in premature infants, there are still some concerns regarding the quality, safety, optimal dosage, and treatment duration of probiotic preparations. Antibiotic prophylaxis does not reduce the incidence of NEC, and prolonged initial empirical use of antibiotics might in fact increase the risk of NEC for high-risk premature infants. Lastly, standardized feeding protocols are strongly recommended, both for prevention of postnatal growth restriction and NEC.

Application of prolonging small feeding volumes early in life to prevent of necrotizing enterocolitis in very low birth weight preterm infants

Objective:To study the effects of prolonging small feeding volumes early in life on the incidence of necrotizing enterocolitis(NEC)in very low birth weight(VLBW)preterm infants.Methods:A total of 128 VLBW infants who could not be breastfed were assigned into the experimental group(63 cases)and the control group(65 cases)using a random number table.The experiment group was fed 12 mL/(kg·d)on day 1 which was increased to 24 mL/(kg·d)for the first 10 study days.The control group was fed 12 mL/(kg·d)for the first 14e48 hours.Then,the feeding volume increased by 24-36 mL/(kg·d)up to 140e160 mL/(kg·d)and maintained until the 10th day after birth.The incidence of feeding intolerance and NEC,duration of hospitalization,time to full enteral feedings,incidence of intrahepatic cholestasis,and the levels of gastrin and motilin in serum were assessed.Results:The incidence of feeding intolerance was significantly lower in the experimental group compared with the control group(15.87% vs.33.84%).There was a significant reduction in the incidence of NEC between the experimental and control groups(7.9% vs.16% in the control group).Conclusion:A protocol that prolongs small feeding volumes early in life can reduce the incidence and severity of NEC,but still warrants further study.

New Frontiers of Necrotizing Enterocolitis: From Pathophysiology to Treatment

Necrotizing enterocolitis [NEC] is an inflammatory disease of intestine largely occuring in preterm infants with a wide range of damage from minimal injury limited to mucosa to extensive necrosis of bowel wall and perforation. Despite advancements in neonatal care, mortality remains high [30% - 50%] and controversy still persists with regards to the most appropriate management of neonates with necrotizing enterocolitis. The main factors thought to be involved in the pathogenesis of NEC are: relatively hyper-reactive state of premature intestine, enteral feeding and bacterial colonization. In this review, we discuss current knowledge about the epidemiology, pathophysiology, imaging, medical and surgical management of necrotizing enterocolitis and describe novel strategies for prevention and treatment.

Mucin 1 and interleukin-11 protein expression and inflammatory reactions in the intestinal mucosa of necrotizing enterocolitis children after surgery

BACKGROUND Necrotizing enterocolitis(NEC)of the newborn is a frequently occurring clinical disease in infants.The mortality rate of NEC in premature infants is as high as 50%,and the morbidity rate is on the rise.NEC has already caused serious impacts on newborn survival and poses serious threats to both children and families.AIM To investigate the expression and significance of mucin 1(MUC1)and interleukin-11(IL-11)in the intestinal mucosa of infants with neonatal NEC after surgery.METHODS Forty-eight postoperative intestinal mucosal specimens from children with NEC(NEC group)and twenty-two intestinal mucosal specimens from children with congenital intestinal atresia(control group)were collected in our hospital.Immunohistochemical staining and Western blot analysis were used to examine the protein expression of MUC-1 and IL-11 in the two groups.The serum levels of tumor necrosis factor-α(TNF-α)and IL-1βin the two groups were measured by enzyme-linked immunosorbent assay,and the relationship between MUC-1 and IL-11 protein expression and serum TNF-αand IL-1βlevels was analyzed by the linear correlation method.RESULTS The protein expression of MUC-1 and IL-11 in the NEC group was significantly lower than that in the control group,and the difference was statistically significant(P<0.05).The levels of serum TNF-αand IL-1βin the NEC group were significantly higher than those in the control group(P<0.05).The protein expression of MUC-1 and IL-11 in the NEC group negatively correlated with serum TNF-αand IL-1βlevels(P<0.05).There was a significant negative correlation between the protein expression of MUC-1 and IL-11 and the levels of serum TNF-αand IL-1βin the NEC group.CONCLUSION The protein expression of MUC1 and IL-11 in the intestinal mucosa of children with NEC is significantly downregulated after surgery.This downregulation may be involved in the pathogenesis of this disease and has a certain correlation with inflammatory response factors in children with NEC.

Neonatal necrotizing enterocolitis caused by umbilical arterial catheter-associated abdominal aortic embolism:A case report

BACKGROUND Reports of necrotizing enterocolitis(NEC)caused by umbilical arterial catheter(UAC)-associated abdominal aortic embolism in neonates are rare.Herein,we report the case of an extremely low birth weight(ELBW)infant with NEC caused by UAC-associated abdominal aortic embolism.CASE SUMMARY A female infant,aged 21 min and weighing 830 g at 28+6 wk of gestational age,was referred to our hospital because of premature birth and shallow breathing.The patient was diagnosed with ELBW,neonatal respiratory distress syndrome,neonatal intrauterine infection,and neonatal asphyxia.Umbilical arterial and venous catheters were inserted on the day after birth and were removed 9 d later,according to the doctor’s plan.Within 48 h after extubation,the patient’s manifestations included poor responsiveness,heart rate range of 175-185/min,and currant jelly stool.Therefore,we considered a diagnosis of NEC.To determine the cause,we used B-mode ultrasound,which revealed a partial abdominal aortic embolism(2 cm×0.3 cm)and abdominal effusion.The patient was treated with nil per os,gastrointestinal decompression,anti-infective therapy,blood transfusion,and low-molecular-weight heparin sodium q12h for anticoagulant therapy(from May 20 to June 1,the dosage of low-molecular-weight heparin sodium was adjusted according to the anti-Xa activity during treatment).On the 67th day after admission,the patient fully recovered and was discharged.CONCLUSION The abdominal aortic thrombosis in this patient was considered to be catheter related,which requires immediate treatment once diagnosed.The choice of treatment should be determined according to the location of the thrombus and the patient’s condition.

Intestinal microbiome changes in an infant with right atrial isomerism and recurrent necrotizing enterocolitis:A case report and review of literature

BACKGROUND Necrotizing enterocolitis(NEC)is a multifactorial disease that predominantly affects premature neonates.Intestinal dysbiosis plays a critical role in NEC pathogenesis in premature neonates.The main risk factor for NEC in term infants is mesenteric hypoperfusion associated with ductaldependent congenital heart disease(CHD)that eventually leads to intestinal ischemia.The incidence of NEC in neonates with critical CHD is 6.8%-13%.However,the role of the intestinal microbiome in NEC pathogenesis in infants with ductal-dependent CHD remains unclear.CASE SUMMARY A male term neonate with right atrial isomerism underwent modified Blalock-Taussig shunt placement on the 14^(th)day of life and had persistent mesenteric hypoperfusion after surgery.The patient had episodes of NEC stageⅡA on the 1^(st)and 28^(th)days after cardiac surgery.Fecal microbial composition was analyzed before and after cardiac surgery by sequencing region V4 of the 16S rRNA gene.Before surgery,species belonging to genera Veillonella and Clostridia and class Gammaproteobacteria were detected,Bifidobacteriaceae showed a low abundance.The first NEC episode was associated with postoperative hemodynamic instability,intestinal ischemiareperfusion injury during cardiopulmonary bypass,and a high abundance of Clostridium paraputrificum(Clostridium sensu stricto I)(56.1%).Antibacterial therapy after the first NEC episode resulted in increased abundance of Gammaproteobacteria,decreased abundance of Firmicutes,and low alpha diversity.These changes in the microbial composition promoted the growth of Clostridium sensu strictoⅠ(72.0%)before the second NEC episode.CONCLUSION A high abundance of Clostridium sensu strictoⅠand mesenteric hypoperfusion may have contributed to NEC in the present case.

Expression of SIRT1 signaling pathway in intestinal tissues of neonatal necrotizing enterocolitis

Objective:To analyze the expression of sirtuin type 1 (SIRT1) signal pathway in intestinal tissues of neonatal necrotizing enterocolitis (NEC), and to preliminarily explore the role of SIRT1 in the occurrence of NEC.Methods: From January 2017 to June 2017, the ileal tissues of 35 neonates with NEC underwent one-stage fistula treatment were selected as NEC group, and the ileal tissues of these 35 neonates underwent two-stage fistula treatment were selected as control group. The expression levels of SIRT1, transcription factor-nuclear factor (NF-κB), and SUMO specific protease 1 (SENP1) were detected by qRT-PCR;the expressions of SIRT1, NF-κB, and SENP1 were detected by immunohistochemistry and Western blotting (WB). Results: SIRT1 mRNA, protein positive expression rate, average optical density, and relative protein expression in intestinal tissues of children in NEC group were significantly lower than those in control group (P<0.05), however, the expression of NF-κB, the SENP1 mRNA and protein positive expression rates, the average optical density, and relative protein expression were significantly increased (P<0.05).Conclusion: SIRT1 is low expressed in intestinal tissues of children with NEC, the possible reason is that SIRT1 signaling pathway is suppressed and then NEC occurs.

Association between Supraventricular Tachycardia and Necrotizing Enterocolitis: A Case-Control Study

Background: Necrotizing enterocolitis (NEC) is the most common and fatal gastrointestinal disease encountered in the Neonatal Intensive Care Unit. Several case reports have shown an association between supraventricular tachycardia and necrotizing enterocolitis. This study aimed to determine the association between supraventricular tachycardia and necrotizing enterocolitis. Methods: This study was conducted from April 1st, 2016 to March 31st, 2022, at the Department of Pediatrics, Zhongnan Hospital of Wuhan University, Hubei, China. The records of 74 subjects with the diagnosis of necrotizing enterocolitis (NEC) were obtained from the hospital’s medical data records. Consequently, 74 gender, gestational age, and birth weight-matched controls (babies without NEC) were recruited as controls. Results: Of the 74 cases, 47.3% of the cases were males, and 52.7% were females. Regarding the birth weight and gestational age, 77% of the cases had low birth weight (LBW) and 86.5% were premature. In terms of Apgar score, 93.2% of NEC cases had an Apgar score of >7 at five minutes. The median values of white blood cells, platelets, and hemoglobin of cases were 10.90 (8.09, 13.80), 227 (169.75, 295.50), and 155.6 (130.53, 170.95), respectively. No Association between supraventricular tachycardia and necrotizing enterocolitis (P = 1.00). Conclusion: No association between necrotizing enterocolitis and supraventricular tachycardia was found. Further multicenter-based studies examining whether there is a potential relationship exists between supraventricular tachycardia and the development of necrotizing enterocolitis are required.

NLRP3 activation in macrophages promotes acute intestinal injury in neonatal necrotizing enterocolitis

Background Macrophages are involved in various immune inflammatory disease conditions.This study aimed to investigate the role and mechanism of macrophages in regulating acute intestinal injury in neonatal necrotizing enterocolitis(NEC).Methods CD68,nucleotide-binding oligomerization domain,leucine-rich repeat,and pyrin domain-containing 3(NLRP3),cysteine aspartate-specific protease-1(caspase-1),and interleukin-1β(IL-1β)in paraffin sections of intestinal tissues from NEC and control patients were detected with immunohistochemistry,immunofluorescence,and western blot.Hypertonic pet milk,hypoxia and cold stimulation were used to establish a mouse(wild type and Nlrp3^(-/-))model of NEC.The mouse macrophage(RAW 264.7)and rat intestinal epithelial cell-6 lines were also cultured followed by various treatments.Macrophages,intestinal epithelial cell injuries,and IL-1β release were determined.Results Compared to the gut“healthy”patients,the intestinal lamina propria of NEC patients had high macrophage infiltration and high NLRP3,caspase-1,and IL-1β levels.Furthermore,in vivo,the survival rate of Nlrp3^(-/-)NEC mice was dramatically improved,the proportion of intestinal macrophages was reduced,and intestinal injury was decreased compared to those of wild-type NEC mice.NLRP3,caspase-1,and IL-1β derived from macrophages or supernatant from cocultures of macrophages and intestinal epithelial cells also caused intestinal epithelial cell injuries.Conclusions Macrophage activation may be essential for NEC development.NLRP3/caspase-1/IL-1β cellular signals derived from macrophages may be the underlying mechanism of NEC development,and all these may be therapeutic targets for developing treatments for NEC.

Development of necrotizing enterocolitis after blood transfusion in very premature neonates

Background Prior studies report conflicting evidence on the association between packed red blood cell(PRBC)transfu-sions and necrotizing enterocolitis(NEC),especially in early weeks of life where transfusions are frequent and spontaneous intestinal perforation can mimic NEC.The primary objective of this study was to evaluate the association between PRBC transfusions and NEC after day of life(DOL)14 in very premature neonates.Methods A retrospective cohort analysis of very premature neonates was conducted to investigate association between PRBC transfusions and NEC after DOL 14.Primary endpoints were PRBC transfusions after DOL 14 until the date of NEC diagnosis,discharge,or death.Wilcoxon ranked-sum and Fisher's exact tests,Cox proportional hazards regression,and Kaplan-Meier curves were used to analyze data.Results Of 549 premature neonates,186(34%)received transfusions after DOL 14 and nine(2%)developed NEC(median DOL=38;interquartile range=32-46).Of the nine with NEC after DOL 14,all were previously transfused(P<0.001);therefore,hazard of NEC could not be estimated.Post hoc analysis of patients from DOL 10 onward included five additional patients who developed NEC between DOL 10 and DOL 14,and the hazard of NEC increased by a factor of nearly six after PRBC transfusion(hazard ratio=5.76,95%confidence interval=1.02-32.7;P=0.048).Conclusions Transfusions were strongly associated with NEC after DOL 14.Prospective studies are needed to determine if restrictive transfusion practices can decrease incidence of NEC after DOL 14.

Hepatoblastoma with neonatal necrotizing enterocolitis:Two case reports

We report two children with hepatoblastoma(HB)with a history of neonatal necrotizing enterocolitis(NEC).Case 1 was diagnosed with HB at 5 months of age.Liver enlargement was found during the NEC operation at 3 months of age and then was clinically diagnosed by imaging.After six chemotherapy courses,a partial hepatectomy was performed.Three months after ceasing the chemotherapy,a chest computed tomography scan suggested that distant metastasis of the tumor should be considered,and the lesion was removed.However,9 months after the operation,alpha-fetoprotein concentrations were increased,and abdominal imaging showed a recurrence of the tumor in situ,resulting in a hepatectomy.Case 2 was diagnosed with NEC shortly after birth and underwent an intestinal resection and anastomosis 1 month later.He was diagnosed with HB at 3 years of age.Hepatectomy was performed after five courses of chemotherapy.Chemotherapy was stopped after 10 courses,and alpha-fetoprotein concentrations were normal.At present,both children have survived and are in a healthy condition.Physicians should be aware of the possibility of HB and a history of NEC in children.Premature birth and low birth weight are common factors leading to the pathogenesis of HB and NEC.The association between these two diseases requires further study。

NEC患儿T抗原暴露导致红细胞多凝集现象的鉴定及输血策略

目的探讨坏死性小肠结肠炎(necrotising enterocolitis,NEC)患儿发生T抗原暴露导致红细胞多凝集现象的鉴定方法,为其制定输血策略,为指导临床为患儿科学、合理用血提供参考依据。方法在本中心进行疑难交叉配血的2例NEC患儿,采用试管法对其进行ABO血型和Rh血型鉴定、直接抗人球蛋白试验(direct antiglobulin test,DAT)、抗体筛查试验及交叉配血试验,采用3例ABO同型献血者血浆,3例AB型献血者血浆,3例同型脐血血浆等方法发现其出现红细胞多凝集现象,并用花生凝集素、MN血型抗体鉴定试剂及凝聚胺方法鉴定其多凝集现象。结果两例患儿红细胞抗体筛查实验阴性;DAT实验阴性;交叉配血实验主侧阴性;次侧盐水法阳性,抗人球蛋白实验阳性;患儿红细胞与献血者同型血浆、AB型血浆呈阳性反应,与同型脐血血浆呈阴性反应;与花生凝集素均呈阳性反应患儿MN血型抗原均为阴性,患儿红细胞加入凝聚胺试剂不出现预期凝集现象。结论NEC患儿出现多凝集红细胞现象表现为DAT实验阴性,但是次侧交叉配血不相合现象。此时应采用多种方法鉴定其红细胞发生多凝集现象,准确识别患儿红细胞多凝集现象,选择合适的血液产品,避免延误患儿输血治疗。

早期应用多重益生菌对早产儿的影响

目的分析出生后早期应用多重益生菌对早产儿临床并发症尤其是坏死性小肠结肠炎(NEC)发生的预防作用及对智力发育的影响。方法选择2022年2月至2023年2月于郑州大学第三附属医院住院治疗的120例胎龄<34周的早产儿,按随机数字表法分成试验组和对照组,各60例。试验组出生后第2天添加多重益生菌,连续28 d;给予对照组正常母乳或配方奶粉喂养。观察早产儿临床并发症发生情况,尤其是NEC全阶段发病率、NEC(≥2期)发病率和智能发育评定分值。结果试验组NEC全阶段发病率低于对照组(P<0.05),两组NEC(≥2期)发病率和其他并发症发生率比较差异无统计学意义(P>0.05);试验组月龄6个月时智能发育评分高于对照组(P<0.05)。试验组全程未发现不良反应。结论早产儿出生后早期应用多重益生菌是安全有效的,可以促进早产儿的智力发育,降低早产儿NEC全阶段发病率,但不能降低NEC发生的严重程度。

血清淀粉样蛋白A在新生儿败血症、坏死性小肠结肠炎早期诊断中的价值

新生儿早/晚发型败血症(EOS/LOS)、坏死性小肠结肠炎(NEC)是新生儿期常见疾病,严重病例预后不良。因早期临床表现不特异、诊断较困难,现非特异性实验室检查(C反应蛋白,降钙素原等)尚存缺点,可能导致漏诊误诊。血清淀粉样蛋白A(SAA)是一种新型急性期反应物,其在EOS/LOS、NEC早期即明显升高、持续时间久,与疾病严重程度相关、且能反映治疗效果,可作为这两种疾病诊治的生物标记物。本文就SAA及其在EOS/LOS、NEC早期诊断中的价值进行综述,为该类疾病诊治提供新的参考。

MD量表联合修正Bell分期对早产儿坏死性小肠结肠炎预后预测价值

目的应用代谢紊乱评分(MD)量表联合修正Bell分期,对坏死性小肠结肠炎的新生儿病例进行回顾性分析,为提高临床治愈率、评估手术时机、改善预后提供依据。方法采用回顾性分析方法,纳入2019年1月至2022年12月本院NICU收治的NEC早产儿224例,根据修正Bell分期分为IB期、ⅡA期、ⅡB期、Ⅲ期;根据患儿是否有手术治疗指征分为手术组与保守组,根据病情转归分为治愈组、预后不良组,分析MD评分及修正Bell分期与早产儿NEC病情危重程度、手术干预时机选择及预后的相关性。结果依据修正Bell分期,IB期83例,ⅡA期52例,ⅡB期47例,Ⅲ期42例,保守组165例,手术组59例,治愈组141例,预后不良组83例,预后不良组(好转79例、死亡4例)。IB期保守治疗,ⅡA期3例手术,ⅡA期保守组和手术组的预后差异不具有统计学意义(P=1.0);ⅡB期选择手术治疗21例,手术治疗治愈率高于保守治疗治愈率,且预后差异具有统计学意义(χ^(2)=6.300,P=0.012)。Ⅲ期选择保守治疗的患儿7例,其中家属放弃治疗后死亡4例,选择手术治疗患儿35例,手术组治愈率高于保守组治愈率,但Ⅲ期保守组与手术组预后差异无统计学意义(χ^(2)=1.577,P=0.209)。单因素分析MD量表各变量与早产儿NEC预后的相关性,出现酸中毒、PCT升高、低钠血症、血小板减少、低血压、中性粒细胞减少与预后不良的相关性具有统计学意义(P<0.05)。影响NEC早产儿预后的多因素Logistic回归分析显示,手术治疗有利预后(β=2.844),酸中毒(OR=0.076,95%CI:0.025~0.232)、血小板减少(OR=0.173,95%CI:0.065~0.463)、手术治疗(OR=17.178,95%CI:4.330~68.142)对预后的影响差异有统计学意义(P<0.01)。MD≥4在手术组和保守组中的分布差异有统计学意义(χ^(2)=109.895,P<0.01),Bell分期≥ⅡB期在手术组和保守组中的分布差异有统计学意义(χ^(2)=101.859,P<0.01)。结论针对NEC早产儿应用修正Bell分期、MD评分,可预测患儿预后情况,为疾病严重程度评估及手术时机选择提供依据。

外周血受体相互作用蛋白激酶3、混合系列蛋白激酶样结构域水平与新生儿坏死性小肠结肠炎病情严重程度的关系

目的分析新生儿坏死性小肠结肠炎(NEC)患儿外周血受体相互作用蛋白激酶3(RIPK3)、混合系列蛋白激酶样结构域(MLKL)的表达情况及其与病情严重程度的关系。方法选取92例NEC患儿纳入NEC组,并根据病情严重程度进一步分为轻度NEC组(Ⅰ级)60例和重度NEC组(Ⅱ~Ⅲ级)32例,另选取同期诊治的60例腹股沟斜疝患儿纳入对照组。采用实时荧光定量聚合酶链反应检测外周血RIPK3 mRNA、MLKL mRNA表达;采用Pearson相关分析法明确NEC组外周血RIPK3 mRNA与MLKL mRNA表达的相关性;采用免疫印迹法检测NEC回肠组织和正常回肠组织中RIPK3、MLKL蛋白表达;采用多因素Logistic回归分析明确重度NEC发生的独立危险因素。绘制受试者工作特征曲线,分析外周血RIPK3 mRNA、MLKL mRNA单独及联合预测重度NEC的价值。结果NEC组外周血RIPK3 mRNA、MLKL mRNA相对表达量分别为(2.41±0.52)、(3.03±0.64),高于对照组的(1.02±0.21)、(0.93±0.20),差异有统计学意义(P<0.001)。NEC回肠组织中RIPK3、MLKL蛋白相对灰度值分别为(1.20±0.21)、(1.13±0.24),高于正常回肠组织的(0.34±0.12)、(0.32±0.11),差异有统计学意义(P<0.05)。NEC组患儿外周血RIPK3 mRNA与MLKL mRNA相对表达量呈正相关(r=0.623,P<0.001)。重度NEC组合并气腹征、多器官功能障碍综合征、败血症者占比和RIPK3 mRNA、MLKL mRNA相对表达量均高于轻度NEC组,差异有统计学意义(P<0.05);RIPK3 mRNA、MLKL mRNA相对表达量升高是重度NEC发生的独立危险因素(P<0.05)。外周血RIPK3 mRNA、MLKL mRNA联合预测重度NEC的曲线下面积大于RIPK3 mRNA、MLKL mRNA单独预测(Z=4.127、4.261,P<0.05)。结论RIPK3 mRNA、MLKL mRNA在NEC患儿外周血中表达升高,两者均与NEC病情严重程度有关,且两者联合检测对重度NEC具有较高的预测价值。