Objective:To investigate the expression and clinical significance of the complement C3 and the S100 calcium binding protein A10 (S100A10) in children with traumatic brain injury (TBI).Methods:This case-control study included 129 TBI children admitted to the Affiliated Xuzhou Children′s Hospital of Xuzhou Medical University from January 2023 to November 2024.The patients were divided into a mild group (85 cases) and a moderate-to-severe group (44 cases).Thirty children with inguinal hernia but no underlying diseases admitted to the hospital during the same period were enrolled as the control group A. Twenty children whose lumbar puncture examination showed normal cerebrospinal fluid results and imaging tests showed no central nervous system disorder were included in the control group B. The children with moderate-to-severe TBI were followed up for 1 month after injury and further divided into good and poor prognosis groups.One-way (repeated-measures) analysis of variance (ANOVA) and t-tests were used to compare differences in complement C3 and S100A10 levels in serum and cerebrospinal fluid among groups.The correlation analysis was performed using the Spearman rank correlation method.Receiver operating characteristic (ROC) curves were drawn to evaluate the value of complements C3 and S100A10 proteins for predicting TBI severity. Results:The serum complement C3 levels in control group A, mild TBI, and moderate-to-severe TBI groups were (1.15±0.26) g/L, (1.02±0.09) g/L, (0.87±0.15) g/L, respectively.The difference in serum complement C3 levels was statistically significant among these three groups ( F=53.661, P<0.001).The serum S100A10 levels in control group A, mild TBI, and moderate-to-severe TBI groups were (0.09±0.03) μg/L, (0.17±0.04) μg/L, (0.32±0.11) μg/L, respectively.The difference in serum S100A10 levels was statistically significant among these three groups ( F=71.093, P<0.001).The levels of complement C3 and S100A10 in the cerebrospinal fluid (30 min post-operation) of children with severe TBI were significantly higher than those in the control group B, with statistically significant differences (all P<0.01).Correlation analysis revealed that Glasgow Coma Scale scores showed a positive correlation with serum complement C3 levels and a negative correlation with S100A10 levels ( r=0.592, -0.705; all P<0.001).The serum complement C3 and S100A10 levels were (0.90±0.13) g/L and (0.30±0.10) μg/L in the good prognosis group, and (0.74±0.16) g/L and (0.42±0.11) μg/L in the poor prognosis group, respectively.Both serum complement C3 and S100A10 levels were statistically significantly different between good and poor prognosis groups ( t=3.025, -3.014; all P<0.01).The complement C3 level in the cerebrospinal fluid of severe TBI children was (0.093±0.007) g/L 30 min after operation, and it gradually increased to reach the first peak at day 3 and the second peak at day 5 postoperatively[(0.112±0.005) g/L and (0.120±0.010) g/L, respectively].The difference in the complement C3 level in the cerebrospinal fluid of severe TBI children was significant between 30 min and 3-5 d after operation ( F=42.756, P<0.01).The S100A10 level in the cerebrospinal fluid of severe TBI children was (2.56±0.31) μg/L 30 min after operation, and then it showed a sustained increase, reaching (4.09±0.13) μg/L at day 7 postoperatively.The difference in the S100A10 level in the cerebrospinal fluid of severe TBI children was significant between 30 min and 7 d after operation ( F=110.676, P<0.01).ROC curve analysis showed that the areas under the curve for predicting moderate-to-severe TBI based on serum complement C3 and S100A10 levels were 0.802 and 0.889, respectively (all P<0.01). Conclusions:Serum complement C3 levels are significantly decreased whereas serum S100A10 levels are markedly elevated in pediatric TBI patients.The measurement of serum complement C3 and S100A10 levels can aid in the clinical assessment of the severity and prognosis of TBI children.Both complement C3 and S100A10 levels in cerebrospinal fluid show a significant elevation within 7 days after operation in severe pediatric TBI, which is potentially linked to sustained astrocyte activation.

Objective:To investigate the expression and clinical significance of the complement C3 and the S100 calcium binding protein A10 (S100A10) in children with traumatic brain injury (TBI).Methods:This case-control study included 129 TBI children admitted to the Affiliated Xuzhou Children′s Hospital of Xuzhou Medical University from January 2023 to November 2024.The patients were divided into a mild group (85 cases) and a moderate-to-severe group (44 cases).Thirty children with inguinal hernia but no underlying diseases admitted to the hospital during the same period were enrolled as the control group A. Twenty children whose lumbar puncture examination showed normal cerebrospinal fluid results and imaging tests showed no central nervous system disorder were included in the control group B. The children with moderate-to-severe TBI were followed up for 1 month after injury and further divided into good and poor prognosis groups.One-way (repeated-measures) analysis of variance (ANOVA) and t-tests were used to compare differences in complement C3 and S100A10 levels in serum and cerebrospinal fluid among groups.The correlation analysis was performed using the Spearman rank correlation method.Receiver operating characteristic (ROC) curves were drawn to evaluate the value of complements C3 and S100A10 proteins for predicting TBI severity. Results:The serum complement C3 levels in control group A, mild TBI, and moderate-to-severe TBI groups were (1.15±0.26) g/L, (1.02±0.09) g/L, (0.87±0.15) g/L, respectively.The difference in serum complement C3 levels was statistically significant among these three groups ( F=53.661, P<0.001).The serum S100A10 levels in control group A, mild TBI, and moderate-to-severe TBI groups were (0.09±0.03) μg/L, (0.17±0.04) μg/L, (0.32±0.11) μg/L, respectively.The difference in serum S100A10 levels was statistically significant among these three groups ( F=71.093, P<0.001).The levels of complement C3 and S100A10 in the cerebrospinal fluid (30 min post-operation) of children with severe TBI were significantly higher than those in the control group B, with statistically significant differences (all P<0.01).Correlation analysis revealed that Glasgow Coma Scale scores showed a positive correlation with serum complement C3 levels and a negative correlation with S100A10 levels ( r=0.592, -0.705; all P<0.001).The serum complement C3 and S100A10 levels were (0.90±0.13) g/L and (0.30±0.10) μg/L in the good prognosis group, and (0.74±0.16) g/L and (0.42±0.11) μg/L in the poor prognosis group, respectively.Both serum complement C3 and S100A10 levels were statistically significantly different between good and poor prognosis groups ( t=3.025, -3.014; all P<0.01).The complement C3 level in the cerebrospinal fluid of severe TBI children was (0.093±0.007) g/L 30 min after operation, and it gradually increased to reach the first peak at day 3 and the second peak at day 5 postoperatively[(0.112±0.005) g/L and (0.120±0.010) g/L, respectively].The difference in the complement C3 level in the cerebrospinal fluid of severe TBI children was significant between 30 min and 3-5 d after operation ( F=42.756, P<0.01).The S100A10 level in the cerebrospinal fluid of severe TBI children was (2.56±0.31) μg/L 30 min after operation, and then it showed a sustained increase, reaching (4.09±0.13) μg/L at day 7 postoperatively.The difference in the S100A10 level in the cerebrospinal fluid of severe TBI children was significant between 30 min and 7 d after operation ( F=110.676, P<0.01).ROC curve analysis showed that the areas under the curve for predicting moderate-to-severe TBI based on serum complement C3 and S100A10 levels were 0.802 and 0.889, respectively (all P<0.01). Conclusions:Serum complement C3 levels are significantly decreased whereas serum S100A10 levels are markedly elevated in pediatric TBI patients.The measurement of serum complement C3 and S100A10 levels can aid in the clinical assessment of the severity and prognosis of TBI children.Both complement C3 and S100A10 levels in cerebrospinal fluid show a significant elevation within 7 days after operation in severe pediatric TBI, which is potentially linked to sustained astrocyte activation.

Objective To investigate the presence of pepsin and trypsin in the middle ear effusion of children with acute suppurative otitis media(ASOM).Methods Middle ear effusion samples were collected from 71 children with ASOM at Children's Hospital of Xuzhou.According to the characteristics of the middle ear effusions,the effu-sion was divided into serous and mucous types.The pH testing,Western Blotting(WB),and enzyme-linked immu-nosorbent assay(ELISA)were performed.Results ① There were 49.29％(35/71)of ASOM patients had a posi-tive RSI score(＞13).② The positive rate of pepsin in ASOM children was 49.29％(35/71),and the positive rate of trypsin was 42.25％(30/71).In addition,the positive rate of pepsin in RSI-positive children was 100％(35/35),and the positive rate of trypsin was 60％(21/35).There was no significant difference in the positive rate of pepsin and trypsin between serous and mucous middle ear effusion(P＞0.05).③ The pepsin concentration was 47.80(39.80,69.30)ng/ml and the trypsin concentration was 291.87±20.45 ng/ml in middle ear effusion of chil-dren with ASOM who had a positive WB test,and the trypsin concentration was significantly higher than pepsin(P＜0.05).There was no significant difference between the pepsin and the trypsin concentrations in serous and mu-cous middle ear effusion(P＞0.05).④ The pH value of mucous middle ear effusion was 7.39±0.28,and the pH value of serous middle ear effusion was 7.36±0.26.There was no significant difference between the pH value in se-rous and mucous middle ear effusion(P＞0.05).Conclusion The detection rates of pepsin and trypsin in middle ear effusion of children with ASOM were high which has important diagnostic value for children with ASOM combined with LPRD.

Objective:To investigate the effect of hypoxia-inducible factor 1α (HIF-1α)/Bcl-2 and adenovirus E1B19kDa-interacting protein 3 (BNIP3)-mediated mitophagy on neuronal apoptosis after traumatic brain injury (TBI).Methods:HT22 cells (mouse hippocampal neurons) were chosen; oxygen-glucose deprivation/re-oxygenation (OGD/R) was used to establish in vitro TBI models. Expressions of HIF-1α and BNIP3 were regulated by HIF-1α, BNIP3-specific small interfering RNA (siRNA) or plasmid vector transfection. Experiment 1 was performed to investigate the effect of BNIP3-mediated mitophagy on neuronal apoptosis after TBI; HT22 cells were divided into 4 groups ( n=3): siRNA control group (normal culture after negative siRNA transfection), siRNA+TBI group (OGD/R after negative siRNA transfection), BNIP3-siRNA group (normal culture after BNIP3-siRNA transfection), and BNIP3-siRNA+TBI group (OGD/R after BNIP3-siRNA transfection); the expressions of mitochondrial autophagy related proteins such as HIF-1α, BNIP3, LC3-II, and P62 were detected by Western blotting, mitochondrial autophagosomes were observed by transmission electron microscopy, apoptosis was detected by TUNEL, and lactate dehydrogenase (LDH) activity in cell supernatant was determined by LDH kit. Experiment 2 was performed to investigate the effect of HIF-1α on BNIP3-mediated mitophagy and neuronal apoptosis after TBI; HT22 cells were divided into 8 groups ( n=3): siRNA control group (normal culture after negative siRNA transfection), siRNA+TBI group (OGD/R after negative siRNA transfection), HIF-1α-siRNA group (normal culture after HIF-1α-siRNA transfection), HIF-1α-siRNA+TBI group (OGD/R after HIF-1α-siRNA transfection), empty plasmid group (normal culture after pcDNA3.1[+] transfection), HIF-1α overexpression group (normal culture after HIF-1α plasmid transfection), empty plasmid+TBI group (OGD/R after empty plasmid transfection), HIF-1α overexpression+TBI group (OGD/R after HIF-1α plasmid transfection); the expressions of HIF-1α, BNIP3, LC3-II, P62, TOMM20 and COX IV, apoptosis rate and LDH activity in neurons of each group were determined. Results:(1) In Experiment 1, compared with siRNA control group, siRNA+TBI group had significantly increased BNIP3 expression and LC3-II/I ratio, significantly decreased P62, TOMM20 and COX IV expressions, and statistically increased apoptosis and LDH activity ( P<0.05); compared with siRNA+TBI group, BNIP3-siRNA+TBI group had significantly decreased BNIP3 expression and LC3-II/I ratio, significantly increased P62, TOMM20, and COX IV expressions, and significantly increased apoptosis and LDH activity ( P<0.05); under projective electron microscope, siRNA+TBI group had increased autophagosomes compared with siRNA control group, while BNIP3-siRNA+TBI group had decreased autophagosomes compared with siRNA+TBI group. (2) In Experiment 2, compared with siRNA+TBI group, HIF-1α-siRNA+TBI group had significantly decreased expressions of HIF-1α and BNIP3, and LC3-II/I ratio, significantly increased expressions of P62, TOMM20 and COX IV, and significantly increased apoptosis and LDH activity ( P<0.05); compared with empty plasmid+TBI group, HIF-1α overexpression+TBI group had statistically higher expressions of HIF-1α and BNIP3, and LC3-II/I ratio, significantly decreased expressions of P62, TOMM20 and COX IV, and significantly decreased apoptosis and LDH activity ( P<0.05). Conclusion:HIF-1α can mitigate TBI-induced neuronal apoptosis via promoting BNIP3-mediated mitophagy.