Objective: To investigate the effects of "liver-smoothing and spirit-regulating" acupuncture on the intestinal flora, lipopolysaccharide (LPS) and the hippocampal toll-like receptor 4 (TLR4)/ transcription factor (NF)-κB signaling pathway in depressive disorder mouse model, and to explore its underlying mechanisms. Methods: Eighteen male SPF-grade C57BL/6J mice were randomly assigned to the control, model, and acupuncture groups using a random number method, with six mice in each group. The depression disorder model was induced in mice from both the acupuncture and model groups using CUMS. The mice in the acupuncture group were treated with acupuncture at the acupoints of "Baihui" (DU20), "Yintang" (DU29), "Hegu" (LI4), and "Taichong" (LR3) on the 15th day of modeling, with a duration of 20 min per session, once per day, for 2 consecutive weeks. Behavioral differences were assessed using the sucrose preference test, open field test, and forced swim test. Hematoxylin-eosin staining was used to observe pathological changes in the hippocampus and colon. The levels of the inflammatory factors interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, and LPS in the hippocampus and colon were measured using Enzyme-linked Immunosorbent Assay. Western blotting was used to detect the expression of TLR4 and NF-κB protein in the hippocampus. Changes in gut microbiota structure and abundance were analyzed by 16 S rDNA sequencing. Results: Compared to the control group, the model group showed reduced sucrose preference rate, time in the center area, and total distance, with an increase in immobility time (P<0.01). Inflammatory pathological changes were observed in the hippocampal CA1 region and colon. The contents of IL-1β, IL-6, TNF-α, and LPS in the hippocampus and colon increased (P<0.01). The protein expression levels of hippocampal TLR4 and NF-κB were increased (P<0.01). The Chao1 index was increased (P<0.01). The relative abundances of Pseudomonadales, Acinetobacter, Moraxellaceae, Solibacillus, Escherichia_shigella, Enterobacteriaceae, Enterobacterales, Dubosiella, and Erysipelottichales were decreased, while the relative abundances of Alloprevotella and gram_negative_bacteriurh_cTPY_13 were increased (P<0.05). The pathways of lipopolysaccharide biosynthesis and pathogenic Escherichia coli infection were upregulated, and the pathway of terpenoid backbone biosynthesis was downregulated (P<0.01). Compared to the model group, the acupuncture group showed increased sucrose preference, time in the center area, and total distance, with a decrease in immobility time (P<0.01). The inflammatory pathological changes in the hippocampal CA1 region and colon were alleviated. The contents of IL-1β, IL-6, TNF-α, and LPS in the hippocampus and colon were reduced(P<0.01). The protein expression levels of hippocampal TLR4 and NF-κB were reduced (P<0.01). The Chao1 index was decreased (P<0.05), and the relative abundances of Dubosiella and Erysipelotrichaceae were increased, while the relative abundance of Rikenellaceae, Alloprevotella, and gram_negative_bacteriuch_cTPY_13 were decreased(P<0.05). The pathways of lipopolysaccharide biosynthesis and pathogenic Escherichia coli infection were significantly downregulated, and the pathway of terpenoid backbone biosynthesis was upregulated (P<0.01). Conclusion " Liver-smoothing and spirit-regulating" acupuncture can improve depressive symptoms in depressive disorder mice, potentially through regulating the LPS and TLR4/NF-κB signaling pathway mediated by intestinal flora, reducing the inflammatory response of the hippocampus, and improving the pathological injury of the hippocampus.

To investigate the incidence of postoperative neuropsychic dysfunction (PND) in Parkinson's disease (PD) patients undergoing deep brain stimulation (DBS) and to analyze its influencing factors.

ObjectiveTo explore the potential mechanism of acupuncture with the method of soothing the liver and regulating the mind in improving depressive disorder. MethodsEighteen C57BL/6J mice were randomly divided into blank group， model group， and acupuncture group， with 6 mice in each group. The model group and the acupuncture group were subjected to depression induction by intraperitoneal injection of lipopolysaccharide （LPS）， while the blank group received an equal volume of normal saline once daily for seven consecutive days. Concurrently， the acupuncture group received "soothing the liver and regulating the mind" acupuncture intervention starting from the first day of modeling， once daily for 14 days； whereas the blank group and the model group were only restrained without acupuncture. The sucrose preference test was used to assess sucrose preference rate， the open-field test to measure center stay time and total travel distance， and the forced swim test to evaluate immobility time. Hematoxylin-eosin （HE） staining was performed to observe hippocampal morphological changes. Enzyme-linked immunosorbent assay （ELISA） was used to detect levels of interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） in hippocampal tissue. Western blot analysis was conducted to examine the protein expression levels of Toll-like receptor 4 （TLR4） and nuclear factor-κB （NF-κB） in the hippocampus. ResultsCompared to the blank group， the model group showed a significant reduction in sucrose preference rate， center stay time， and total travel distance， along with a significant increase in immobility time in the forced swim test， hippocampal IL-1β， IL-6， and TNF-α levels， as well as TLR4 and NF-κB protein expression （P＜0.01）， and the histological examination revealed blurred hippocampal neuronal boundaries， loose arrangement， and some neurons exhibiting nuclear pyknosis and deep staining. Compared to the model group， the acupuncture group demonstrated a significant increase in sucrose preference rate， center stay time， and total travel distance， along with a significant reduction in immobility time in the forced swim test， hippocampal IL-1β， IL-6， and TNF-α levels， and TLR4 and NF-κB protein expression （P＜0.01）， and the histological analysis showed that hippocampal neurons in the acupuncture group were more tightly arranged， with reduced nuclear pyknosis and deep staining. ConclusionAcupuncture with the "soothing the liver and regulating the mind" method can significantly improve depression-like behavior， potentially by inhibiting the hippocampal TLR4/NF-κB signaling pathway and alleviating inflammatory responses.

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 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.

Objective::Percutaneous coronary intervention is one of the most common procedures used for the invasive treatment of patients with coronary heart disease; the incidence of in-stent restenosis (ISR) after percutaneous coronary intervention is 5% to 15%. In this study, a competitive endogenous RNA (ceRNA) network was constructed to investigate potential mechanisms involved in ISR.Methods::The expression data for differentially expressed microRNAs (DEmiRNAs) and messenger RNAs (mRNAs) between patients with and without ISR were obtained using limma package. Long noncoding RNAs (lncRNAs) were predicted based on the DEmiRNAs using the miRDB, miRTarBase, and TargetScan databases. An ISR-specific ceRNA network was subsequently constructed and investigated. To verify the key miRNAs of ceRNA, patients with and without ISR were enrolled from Guangdong Provincial Hospital of Chinese Medicine between January 2017 and December 2018 ( n = 8, respectively); plasma was collected from all enrolled patients. Results::Based on the raw data obtained from the Gene Expression Omnibus database, 472 DEmiRNAs and 304 differentially expressed messenger RNAs between patients with and without ISR were identified. A ceRNA network was constructed by combining 270 lncRNAs, 3 miRNAs (miR-125, miR-140, and miR-206), and 4 mRNAs (STRADB, TKT, PCTP, and BTG2). The hub genes of the ceRNA network of ISR included the following: miR-125, miR-206, miR-140, PCDHB9, CASC2, BAK1P1, CSPG4P3Y, CSPG4P4Y, STRCP1, and GRIP2. Verification of miRNAs of ceRNA also showed that the expression of miR-206 was upregulated in patients with ISR vs. those without ISR ( P < 0.05). In contrast, the expression of miR-140 and miR-125 was downregulated in patients with ISR vs. those without ISR ( P < 0.05). Conclusions::This study constructed noncoding RNA-related ceRNA networks for ISR. The results indicated that miR-206, miR-125, and miR-140 may be biomarkers of ISR.

Objective::Percutaneous coronary intervention is one of the most common procedures used for the invasive treatment of patients with coronary heart disease; the incidence of in-stent restenosis (ISR) after percutaneous coronary intervention is 5% to 15%. In this study, a competitive endogenous RNA (ceRNA) network was constructed to investigate potential mechanisms involved in ISR.Methods::The expression data for differentially expressed microRNAs (DEmiRNAs) and messenger RNAs (mRNAs) between patients with and without ISR were obtained using limma package. Long noncoding RNAs (lncRNAs) were predicted based on the DEmiRNAs using the miRDB, miRTarBase, and TargetScan databases. An ISR-specific ceRNA network was subsequently constructed and investigated. To verify the key miRNAs of ceRNA, patients with and without ISR were enrolled from Guangdong Provincial Hospital of Chinese Medicine between January 2017 and December 2018 ( n = 8, respectively); plasma was collected from all enrolled patients. Results::Based on the raw data obtained from the Gene Expression Omnibus database, 472 DEmiRNAs and 304 differentially expressed messenger RNAs between patients with and without ISR were identified. A ceRNA network was constructed by combining 270 lncRNAs, 3 miRNAs (miR-125, miR-140, and miR-206), and 4 mRNAs (STRADB, TKT, PCTP, and BTG2). The hub genes of the ceRNA network of ISR included the following: miR-125, miR-206, miR-140, PCDHB9, CASC2, BAK1P1, CSPG4P3Y, CSPG4P4Y, STRCP1, and GRIP2. Verification of miRNAs of ceRNA also showed that the expression of miR-206 was upregulated in patients with ISR vs. those without ISR ( P < 0.05). In contrast, the expression of miR-140 and miR-125 was downregulated in patients with ISR vs. those without ISR ( P < 0.05). Conclusions::This study constructed noncoding RNA-related ceRNA networks for ISR. The results indicated that miR-206, miR-125, and miR-140 may be biomarkers of ISR.

Objective: To explore the utility of pharyngeal pH monitoring which positive standard is Ryan index in diagnosis of laryngopharyngeal reflux disease. Methods: In a retrospective study, clinical data of 590 patients who had symptoms laryngopharyngeal reflux disease from February 2016 to March 2017 were analyzed. All patients were received electronic laryngoscopy, assessment of reflux symptom index(RSI) and reflux finding score(RFS), and pharyngeal pH monitoring. SPSS 19.0 software was used to analyze the date. Results: There were 94 patients whose Ryan index were positive(15.93%). Among the 94 patients, 70 were positive during upright, 12 during supine and 12 during both upright and supine. There were 40 patients(6.78%)with pH decline events related to symptoms, while those Ryan index were normal. There were 536(90.85%), 417(70.68%), 233(39.49%) and 117(19.83%) patients with pH<6.5, pH<6.0, pH<5.5 and pH<5.0 events respectively. The positive rate of RSI, RFS, RSI and RFS, RSI or RFS were 44.24%, 16.78%, 7.12%, 53.90% respectively. The RFS score in Ryan index positive group was higher than that in Ryan index negative group[(8.2±2.4) vs (4.0±2.9), u=5.424, P<0.05], while the RSI score in Ryan index positive group was not statistically different from that in Ryan index negative group[(11.3±6.2) vs (12.7±5.8), t=1.247, P=0.167]. Conclusions Pharyngeal pH monitoring is an objective and non-invasive method which can reflect laryngopharyngeal reflux directly. However, with the Ryan index as a criterion for the diagnosis of laryngopharyngeal reflux disease, partial patients may be missed. Further studies are needed to obtain more accurate and objective laryngopharyngeal pH statistical index for diagnosis of laryngopharyngeal reflux disease.

Objective]To study the feasibility of Cartilage engineering using fibrin gel and chondrocyte cell sheets.[Methods]rabbit auricular chondrocytes were isolated and cultured to form cell sheets in flasks. The cell sheets were harvested using cell scrapers,and cut into fragments. The two precursor solutions of Fibrin gel were used to suspend the cell sheet fragments and isolated chondrocytes,and then added into the wells of a 48-well plate to form Gelatinous chondroid disc constructs. After in vitro culture, the constructs were implanted into nude mice. After 8 weeks,the constructs were harvested,and the specimens were evaluated using grossly observing, histological and immunohistochemical observation. [Results]Mature cartilage discs were obtained. The histomorphology of the explanted discs appeared non-uniform cartilaginous tissue comprise of regenerated cartilage islands with different size and irregular shape. Immunohistochemistry staining demonstrated that type II collagen highly expressed in the ECM of the cartilage islands. In 1 of the 8 discs,partial ossification was observed.[Conclusion]Fibrin gel is a favourable carrier. Artificial cartilage with stereochemical structure was constructed via combining the fibrin gel and chondrocyte cell sheets.