Objective To observe the efficacy,safety and defecation function of laparoscopic modified Soave short muscle sheath in the treatment of children with Hirschcolon(HD).Methods A total of 91 children with HD admitted to our hospital from January 2018 to May 2023 were selected for the study,and were divided into observation group(modified laparoscopic Soave short muscle sheath operation,41 cases)and control group(traditional laparoscopic Soave Hirschacolon radical resection,50 cases)according to random number table method.Operation time,postoperative blood loss,postoperative defecation time,postoperative hospital stay,length of resection intestine and preoperative enema time were compared between the two groups.White blood cell count(WBC)and C-reactive protein(CRP)levels before and after operation were compared between the two groups,and pain scores were compared between the two groups 12 h,24 h,36 h and 48 h after operation.Postoperative defecation function and postoperative complications were compared between the two groups.Results The operation time[(134.07±22.08)min],postoperative blood loss[(5.17±0.87)ml],postoperative defecation time[(2.10±0.32)d],postoperative hospital stay[(7.59±1.25)d],length of intestinal tube resection[(15.24±2.25)cm]and preoperative enema time[(13.61±2.14)min]in observation group were all lower than those in control group[(159.78±25.44)min,(7.61±1.41)ml,(2.46±0.53)d,(10.59±1.84)d,(16.67±2.54)cm and(16.44±2.57)min](P＜0.05).After operation,the serum WBC and CRP levels of the two groups were significantly increased,but the serum WBC and CRP levels of the observation group were lower than those of the control group(P＜0.05).The pain scores of the observation group(3.42±0.69,3.17±0.64,2.52±0.58,2.06±0.53)at 12 h,24 h,36 h and 48 h were lower than those of the control group(4.47±0.76,3.78±0.72,3.31±0.66,2.83±0.64)(P＜0.05).There was no significant difference in defecation function between the two groups(P＞0.05).The total incidence of postoperative complications was 17.07％in the observation group,which was lower than 34.00％in the control group(P＜0.05).Conclusion The effect of laparoscopic modified Soave short muscle sheath in HD children is better,which can effectively improve various clinical indicators and defecation function,relieve postoperative pain,reduce the degree of infection,reduce postoperative complications,and have higher safety.

Objective:To explore the effect of pediatric critical illness score (PCIS), pediatric risk of mortality Ⅲ score (PRISM Ⅲ), pediatric logistic organ dysfunction 2 (PELOD-2), pediatric sequential organ failure assessment (p-SOFA) score and Glasglow coma scale (GCS) in the prognosis evaluation of septic-associated encephalopathy (SAE).Methods:The data of children with SAE admitted to the Pediatric Intensive Care Unit (PICU), Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences from January 2010 to December 2020 were retrospectively analyzed. They were divided into the survival and death groups according to the clinical outcome on the 28th day after admission. The efficiency of PCIS, PRISM Ⅲ, PELOD-2, p-SOFA and GCS scores for predicting death were evaluated by the area under the ROC curve (AUC). The Hosmer-Lemeshow goodness-of-fit test assessed the calibration of each scoring system.Results:Up to 28 d after admission, 72 of 82 children with SAE survived and 10 died, with a mortality rate of 12.20%. Compared with the survival group, the death group had significantly lower GCS [7 (3, 12) vs. 12 (8, 14)] and PCIS scores [76 (64, 82) vs. 82 (78, 88)], and significantly higher PRISM Ⅲ [14 (12, 17) vs. 7 (3, 12)], PELOD-2 [8 (5, 13) vs. 4 (2, 7)] and p-SOFA scores [11 (5, 12) vs. 6 (3, 9)] ( P<0.05). The AUCs of PCIS, PRISM Ⅲ, PELOD-2, p-SOFA and GCS scores for predicting SAE prognosis were 0.773 ( P=0.012, AUC>0.7), 0.832 ( P=0.02, AUC>0.7), 0.767 ( P=0.014, AUC>0.7), 0.688 ( P=0.084, AUC<0.7), and 0.692 ( P=0.077,AUC<0.7), respectively. Hosmer-Lemeshow goodness-of-fit test showed that PCIS ( χ2=5.329, P=0.722) predicted the mortality and the actual mortality in the best fitting effect, while PRISM Ⅲ ( χ2=12.877, P=0.177), PELOD-2 ( χ2=8.487, P=0.205), p-SOFA ( χ2=9.048, P=0.338) and GCS ( χ2=3.780, P=0.848) had poor fitting effect. Conclusions:The PCIS, PRISM Ⅲ and PELOD-2 scores have good predictive ability assessing the prognosis of children with SAE, while the PCIS score can more accurately evaluate the fitting effect of SAE prognosis prediction.

Background:Circular RNAs(circRNAs)are covalently closed single-stranded RNAs with multiple biological functions.CircRNA.0007127 is derived from the carbon catabolite repression 4-negative on TATA-less(CCR4-NOT)complex subunit 2(CNOT2),which was found to regulate tumor cell apoptosis through caspase pathway.Methods:Potential circRNA.0007127 target microRNAs(miRNAs)were analyzed by miRanda,TargetScan,and RNAhybrid software,and the miRNAs with binding sites for apoptosis-related genes were screened.The roles of circRNA.0007127 and its downstream target,microRNA(miR)-513a-5p,were validated by quantitative real-time polymerase chain reaction(qPCR),flow cytometry,mitochondrial membrane potential,immunofluorescence,western blot,and caspase-8(CASP8)protein activity in vitro in H2O2-induced K-562 cells.The circRNA.0007127-miR-513a-5p and CASP8-miR-513a-5p interactions were verified by luciferase reporter assays.Results:Silencing circRNA.0007127 decreased cell apoptosis by inhibiting CASP8 pathway activation in K-562 cells.Compared with the control group,the expression of CASP8 was reduced by 50％and the 43-kD fragment of CASP8 protein was significantly reduced(P≤0.05).The luciferase reporting assay showed that circRNA.0007127 combined with miR-513a-5p or CASP8,with extremely significant differences(P≤0.001).The overexpression of miR-513a-5p inhibited the gene expression level of CASP8 in a human myeloid leukemia cell model(75％change)and the level of a 43-kD fragment of CASP8 protein(P≤0.01).The rescue experiment showed that cotransfection with circRNA.0007127 small-interfering RNA(siRNA)and the miR-513a-5p inhibitor increased CASP8 gene expression and the apoptosis rate,suggesting that the miR-513a-5p inhibitor is a circRNA.0007127 siRNA antagonist.Conclusions:CircRNA.0007127 regulates K-562 cell apoptosis through the miR-513a-5p/CASP8 axis,which can serve as a novel powerful molecular target for K-562 cells.

Objective:To investigate the effect of melatonin (MEL) influence on lipopolysaccharide (LPS)-induced long-term anxiety-like behavior and activation of astrocytes in septic neonatal rats.Methods:Sprague-Dawley rats were randomly(random number) assigned to the control group, LPS group and LPS+MEL group. Sepsis model was intraperitoneally injected with LPS (1 mg/kg), and neonatal rats in the MEL group were administered with MEL (10 mg/kg) 30 min after LPS injection. At different time points after injection, rats in each group were divided into three subgroups: 3 d, 7 d and 28 d. The expression of GFAP and TNF-α in the corpus callosum was detected by immunofluorescence staining and Western blot. Open-field test was applied to observe anxiety-like behaviors. In vitro, cultured neonatal SD rat astrocytes were divided into the control group, LPS group, LPS+MEL group, and LPS+MEL+luzindole group. Immunofluorescence staining was used to observe the expression of GFAP and TNF-α. Expression of GFAP, TNF-α, p-NF-κBp65, NF-κBp65 protein in astrocytes were assessed by Western blot. RT-qPCR was used to investigate the mRNA expression of GDNF and BDNF. One-way ANOVA and two-way ANOVA were used for comparison of multiple groups of variables. A P<0.05 was considered statistically significant. Results:LPS reduced the duration of movement in the central area and distance in the central area/total distance in open-field test, while melatonin evidently reversed the LPS-induced anxiety-like behavior. Compared with the LPS group, the expressions of GFAP and TNF-α were significantly decreased in the corpus callosum at 3 d and 7 d in the MEL group ( P< 0.05). Compared with the LPS group, MEL could significantly decrease the expression of GFAP, TNF-α and p-NF-κBp65 in astrocytes ( P< 0.05), which could be blocked by Luzindole. In addition, compared with the LPS group, MEL pretreatment could reverse the down regulation of GDNF and BDNF induced by LPS ( P<0.05). Conclusions:MEL can relieve LPS-induced long-term anxiety-like behavior in septic neonatal rats. The mechanism may be related to the inhibition of astrocyte activation and inflammatory reaction through NF - κ B pathway.

Objective:To investigate the effect of melatonin on oligodendrocyte maturation and differentiation in corpus callosum of septic neonatal rats induced by systemic lipopolysaccharide (LPS) injection.Methods:Sprague-Dawley rats were randomly allocated into the control group, septic experimental group, and melatonin group. In the septic experimental group, rats were intraperitoneally administrated with lipopolysaccharide (LPS) (1 mg/kg). In the melatonin group, melatonin was intraperitoneally administered (10 mg/kg) at 0.5 h after LPS injection. The expression level of IL-6, olig1, olig2, and the MAG protein were detected by Western blot at different time points in the three groups. BV-2 cells were used in vitro. For drug administration, the effect of LPS, melatonin and melatonin receptor antagonist, luzindole, on IL-6 expression in BV-2 microglia cell was determined by Western blot. The medium of BV2 cell were collected to treat primary OPCs. The expression level of olig1, olig2 and MAG protein in primary OPCs were detected by Western blot. SPSS 20.0 statistical software was used for analysis, and the data were analyzed by one-way ANOVA and two-way ANOVA. Differences were considered to be statistically significantly if P<0.05. Result:Compared with the LPS group, the expression of IL-6 was significantly decreased in the corpus callosum at 6 h, l d, and 3 d in the melatonin group ( P<0.05). The expression of olig1, olig2 and MAG protein were increased at day 7, 14, and 28 in the melatonin group compared with the LPS group ( P<0.05). In vitro the expressions of IL-6 was significantly increased after LPS treatment ( P<0.05), but was decreased in the LPS+melatonin treatment group ( P<0.05). After treatment with melatonin receptor inhibitor, luzindole, the expressions level of IL-6 was increased ( P<0.05). The expression of olig1, olig2 and MAG protein were decreased with conditioned medium in the LPS BV2 cell group than the control group in the primary OPCs ( P<0.05). However, those were increased with conditioned medium in the LPS+melatonin BV2 cell group than the LPS group ( P<0.05). Conclusions:Melatonin may inhibit the inflammation response in the corpus callosum through its receptor, and may promote the maturation and differentiation of oligodendrocyte, suggesting that melatonin may have therapeutic effect on neuroinflammation and axonal hypomyelination on PWM in septic neonatal rats.

Objective To investigate the effect of interleukin-1β (IL-1β) on the expression of synaptic protein SNAP-25 in the hippocampus in septic neonatal rat induced by systemic lipopolysaceharide (LPS) injection.Methods Sprague-Dawley (SD) rats were randomly divided into two groups:control group and sepsis group.The rat model of sepsis was produced by intraperitoneal injection of 1 mg/kg LPS,and rats in the control group were injected with an equal volume of 0.01 mol/L phosphate buffered saline (PBS).The expression levels of IL-1β and IL-1R1 in the hippocampus at 1,2 and 3 d,and synaptosomal-associated protein 25 (SNAP-25) at 7,14 and 24 d after LPS intraperitoneal injection were detected by Western blot.After cultured for 24 h,primary hippocampal neurons were divided into four groups including the control group,IL-1β (40 ng/mL) treatment group,IL-1β (40 ng/mL) + IL-1Ra (40 ng/mL) treatment group,and IL-1Ra (40 ng/mL) treatment group.The effect of IL-1β on SNAP-25 expression in primary hippocampal neuron was determined by Western blot and real-time PCR.The purity of hippocampal neurons were identified by NeuN immunofluorescence staining and the activity of neurons were detected by CCK-8 assay.All data were analyzed by SPSS version 22.0.The data were analyzed by student-t test and Dunnett-t test.The interaction effects were analyzed by factorial ANOVA.Differences were considered to be statistically significant if P＜ 0.05.Results Compared with the control group,the expressions of IL-1β and IL-1R1 were significantly increased in the hippocampus at 1,2 and 3 d after intraperitoneal injection of LPS (P＜0.05).The expression of SNAP-25 protein was decreased at 7,14,and 28 d after intraperitoneal injection of LPS (P＜0.05).The purity of primary neurons was about up to 92％.The activity of primary neurons was not relatively changed after treated with IL-1β at a dose less than 40 ng/mL.The level of SNAP-25 protein was obviously decreased in primary neurons at 24 h after IL-1β treatment (P＜0.05).IL-1Ra treatment might reverse the effect of IL-1β on primary neurons (P＜0.05).While,the expression of SNAP-25 mRNA was not statistically different in each group (P＞0.05).Conclusions IL-1β may possibly inhibit the expression level of SNAP-25 protein in the hippocampus in the septic rats through its receptor IL-1R1,which would contribute to cognitive dysfunction of septic neonatal rats in later life.

Objective: To explore the effects of allogeneic bone marrow mesenchymal stem cells (BMSCs) on polarization of peritoneal macrophages isolated from rats with sepsis induced by endotoxin/lipopolysaccharide (LPS). Methods: (1) BMSCs were isolated, cultured and purified from 5 SD rats with whole bone marrow adherent method. The third passage of cells were collected for morphologic observation, detection of expressions of stem cell surface markers CD29, CD44, CD45, and CD90 with flow cytometer, and identification of osteogenic and adipogenic differentiation. (2) Another 45 SD rats were divided into sham injury group (SI, n=5), LPS control group (LC, n=20), and BMSCs-treated group (BT, n=20) according to the random number table. Rats in groups LC and BT were injected with LPS (5 mg/kg) via tail vein to induce sepsis; rats in group SI were injected with the same amount of normal saline to simulate the damage. At post injury hour (PIH) 1, rats in group BT were given 1 mL BMSCs (2×10⁶/mL) via tail vein injection; rats in another two groups were injected with equal volume of phosphate buffer saline. Five rats in group SI at PIH 24 and in groups LC and BT at PIH 6, 12, 24, and 48 were sacrificed to harvest lung tissue for pathological observation with HE staining. In addition, rats in group SI at PIH 24 and in groups LC and BT at PIH 24 and 48 were simultaneously performed with intraperitoneal injection of low-glucose DMEM. Then peritoneal fluid was harvested to culture peritoneal macrophages. Flow cytometer was used to assess the positive expression of cell makers of macrophages including CD68 (making gate), CD11c, and CD206 in group SI at PIH 24 and in groups LC and BT at PIH 24 and 48. Data were processed with one-way analysis of variance and LSD test. Results: (1) The third passage of cells showed uniform fiber-like shape similar to fibroblasts. These cells showed positive expressions of CD29, CD44, CD90 and weak positive expression of CD45. They were able to differentiate into osteoblasts and adipocytes. These cells were identified as BMSCs. (2) At PIH 24, the structure of pulmonary alveoli of rats in group SI was clear and complete with no congestion or inflammatory cell infiltration. At PIH 6, the structure of pulmonary alveoli of rats in groups LC and BT was clear with a small amount of inflammatory cell infiltration, slight congestion and pulmonary interstitial thickening. At PIH 12, the inflammatory responses in lung tissue of rats in group LC were more severe than those in group BT with a large amount of inflammatory cell infiltration, serious congestion, and obvious pulmonary interstitial thickening. The pathological results of rats in group BT at PIH 12 was consistent with the results at PIH 6. At PIH 24, the pathological results of rats in groups LC and BT were similar to the results at PIH 12. At PIH 48, the structure of pulmonary alveoli tissue of rats in group LC was still severely disrupted, with a large number of inflammatory cell infiltration and congestion in lung tissue, but pulmonary interstitial thickening was slightly alleviated than before. The condition of rats in group BT nearly recovered to that in group SI. (3) At PIH 24, the positive expression rate of CD11c in peritoneal macrophages of rats in group LC [(83±10)%] was close to that in group BT [(87±7)%, P>0.05], and they were both significantly higher than the rate in group SI [(55±12)%, with P values below 0.01]. The positive expression rate of CD11c in peritoneal macrophages of rats in group LC [(59±11)%] at PIH 48 was close to that in group SI at PIH 24 (P>0.05), and they were both significantly higher than the rate in group BT [(20±11)%] at PIH 48 (with P values below 0.01). At PIH 24, the positive expression percentages of CD206 in peritoneal macrophages of rats were similar among the three groups (with P values above 0.05). The positive expression percentage of CD206 in peritoneal macrophages of rats in group SI at PIH 24 was close to that in group BT at PIH 48 (P>0.05), and they were both significantly lower than the percentage in group LC at PIH 48 (with P values below 0.01). Conclusions BMSCs can reduce the pathological inflammatory responses in the lung of rats with sepsis and inhibit peritoneal macrophages from polarizing into M1 phenotype, whereas they can not promote macrophages to polarize into M2 phenotype.

Objective To investigate microbial characteristics and predisposing factors in gram-negtive bacteria blood stream infection. Methods A descriptive retrospective study was conducted. Patients diagnosed as sepsis with blood culture of G- bacilli and without sepsis were enrolled. The patients were all admitted to ICUs of Guangdong General Hospital from October, 2012 to December, 2014. The clinical characteristics and outcomes were compared. Multiple logistic regression was used to analyse the predisposing factors for sepsis of G- bacilli. Results A total of 148 patients suffered from sepsis of G-bacilli including Acinetobacter baumannii, Escherichia coli and Klebsiella pneumoniae were enrolled. Single-factor analysis showed that patients with sepsis of G- bacilli infection had older ages, higher incidence of coronary heart diseases or congestive heart failure, cerebrovascular diseases or chronic renal insufficiency, hypertension, also higher incidence of longer length of hospital stay before blood was drawn for culture, and higher incidence using of vasoactive agents and pre-admission intravenous antibiotics and lower plasma albumin level (P < 0.05). Conclusions Coronary heart disease or congestive heart failure, chronic renal insufficiency and pre-admission intravenous antibiotics were independent predisposing factors for sepsis of G-bacilli.

Objective To explore whether hypertonic saline would partake in regulating Notch signaling in microglia in experimentally induced cerebral ischemic rats.Methods Male SD rats were randomly divided into sham group, cerebral ischemia group, normal saline group ( NS group ) , 10%hypertonic saline group (10%HS group) , the model of cerebral ischemia were established in all rats except the sham group by using middle cerebral artery occlusion ( MCAO) .After 2 hours of MCAO, the rats were through reperfusion for 24 h.In addition, rats in the normal saline group and 10% HS group were respectively treated with a continuous intravenous injection of normal saline (0.3 mL/h) and 10%HS (0.3 mL/h) by tail vein for 24 h.Immunofluorescence methods, RT-PCR and Western blot were used to detect the expression of Notch1 and intracellular Notch receptor domain ( NICD) .All data was analyzed by one-way analysis of variance ( ANOVA) , The intergroup comparisons were analyzed by the least-significant-difference (LSD) tests.Differences were considered statistically significant if P<0.05.Results Immunofluorescence showed that the expression of Notch1 and NICD were significantly increased in the microglia around peri-ischemia area in cerebral ischemia group and normal saline group compared to sham group;the expression of Notch1 and NICD in the microglia around peri-ischemia area were significantly reduced in 10% HS group compared to ischemia group and NS group.RT-PCR showed that the mRNA expression of Notch1 was significantly increased in ischemia group and NS group compared to sham group ( sham group: 1.000 ± 0.076; ischemia group: 2.203 ±0.283; NS group: 1.616 ±0.185; P <0.01 ); however, it was significantly reduced in 10% HS group compared to ischemia group and NS group ( ischemia group:2.203 ±0.283; NS group: 1.616 ±0.185; 10%HS group: 1.202 ±0.177; P <0.05 ) .Western blot showed that the protein expression of Notch1 was significantly increased in ischemia group and NS group compared to sham group ( sham group: 0.290 ±0.079; ischemia group: 0.750 ±0.029; NS group:0.765 ±0.182;P<0.01);but was significantly reduced in 10%HS group compared to ischemia group and NS group ( ischemia group:0.750 ±0.029; NS group:0.765 ±0.182;10%HS group:0.390 ±0.195;P<0.05 ) .The protein expression of NICD was significantly increased in ischemia group and NS group compared to sham group ( sham group: 0.401 ±0.196; ischemia group: 0.906 ±0.359; NS group:0.847 ±0.153;P<0.01);but was significantly reduced in 10%HS group compared to ischemia group and NS group ( ischemia group:0.906 ±0.359; NS group:0.847 ±0.153;10%HS group:0.561 ±0.165;P<0.05 ) .Conclusion Our results suggest that HS markedly suppresses Notch signaling in microglia around the ischemia tissue area in experimental induced cerebral ischemic rats.

Objective To explore the advantages and disadvantages of helicopter emergency medical services of South China in the long-distance transport for critical patients.Methods A total of 30 patients who received helicopter emergency medical services by Guangdong Generral Hospital from August 2004 to December 2014 were selected as the observation group,and the other 30 patients with similar conditions who received ground emergency medical services were selected as the control group.To analyses the difference between the two groups in the disease,transport distance,transportation time,costs and compliction by χ2-test,t-test and nonparametric test according types of data.Results There were significantly difference between two groups in transport distances (km) [578.0 (313.0,707.5)vs.214.5 (101.5,313.5),P <0.05],set-up time (min) [95.7 (56.7,133.4)vs.10.7 (6.8,15.7),P <0.05],transportation time (min) [112.3 (64.3,152.4) vs.146.8 (67.8,217.5),P <0.05],costs (yuan/h) [14378.5 (9887.0,16348.5)vs.557.0 (356.5, 787.5),P <0.05]and the distance/total time value [2.8 (1.3,4.8)vs.1.4 (0.8,2.8),P <0.05]. There was no significantly difference in the incidence of complications between two groups (χ2 =0.058,P >0.05).Conclusions Helicopter emergency medical services could shorten the transportation time of critical patients on long distance transportation,and improve the efficiency of first-aid.However,there were many disadvantages that need to be improved in the helicopter emergency medical service of China.