ObjectiveTo clarify whether METTL14 mediates the core role of acupuncture at Neiguan (PC6) in promoting myelination and improving behavior in young autistic rats through gene intervention technology. MethodsThe ASD model was established by intraperitoneal injection of valproic acid (VPA) in pregnant rats. Male offspring were intracerebroventricularly injected with adenovirus-packaged METTL14 shRNA (sh-METTL14) or its control (sh-NC) on postnatal day 1, with a model group set as well. Subsequently, the juvenile rats were divided into model group, acupuncture group, acupuncture+sh-NC group, and acupuncture+sh-METTL14 group. The acupuncture group received acupuncture at Neiguan (PC6) from postnatal day 7, once daily for 21 consecutive days. Neurobehavioral changes were evaluated by behavioral tests; METTL14 knockdown efficiency and the expression of METTL14, METTL3, and PTEN were detected by quantitative real-time PCR (qRT-PCR) and Western blot (WB); PTEN m⁶A levels were measured by RNA immunoprecipitation-qPCR (RIP-qPCR); myelin ultrastructure, expression of myelin basic protein (MBP) and neurofascin 155 (NF155), and dendritic spine density were observed using transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), immunofluorescence, qRT-PCR, and primary neuron culture. ResultsBehaviorally, knockdown of METTL14 significantly counteracted the beneficial effects of acupuncture in improving self-grooming, open field exploration, three-chamber social interaction, and Morris water maze learning and memory (P<0.05, P<0.01). Compared with the acupuncture+sh-NC group, the acupuncture+sh-METTL14 group showed significantly decreased mRNA and protein expression of hippocampal METTL14 (P<0.01), and the upregulating effects of acupuncture on METTL3 and PTEN expression were reversed (P<0.01). Meanwhile, knockdown of METTL14 significantly inhibited the acupuncture-induced increase in PTEN m⁶A levels (P<0.01). Morphologically, knockdown of METTL14 attenuated the improvement of myelin structure by acupuncture, reversed the downregulation of MBP and upregulation of NF155 induced by acupuncture, and blocked the increase in dendritic spine density (P<0.05, P<0.01). ConclusionMETTL14 is a key molecule mediating the therapeutic effect of acupuncture at Neiguan. Acupuncture at Neiguan upregulates METTL14, thereby enhancing m⁶A methylation modification of PTEN mRNA to stabilize its expression, ultimately promoting myelin development and improving behavioral symptoms in ASD juvenile rats. This preliminarily reveals the modern biological connotation of “opening Xuanfu and dredging myelin”.

ObjectiveTo clarify whether METTL14 mediates the core role of acupuncture at Neiguan (PC6) in promoting myelination and improving behavior in young autistic rats through gene intervention technology. MethodsThe ASD model was established by intraperitoneal injection of valproic acid (VPA) in pregnant rats. Male offspring were intracerebroventricularly injected with adenovirus-packaged METTL14 shRNA (sh-METTL14) or its control (sh-NC) on postnatal day 1, with a model group set as well. Subsequently, the juvenile rats were divided into model group, acupuncture group, acupuncture+sh-NC group, and acupuncture+sh-METTL14 group. The acupuncture group received acupuncture at Neiguan (PC6) from postnatal day 7, once daily for 21 consecutive days. Neurobehavioral changes were evaluated by behavioral tests; METTL14 knockdown efficiency and the expression of METTL14, METTL3, and PTEN were detected by quantitative real-time PCR (qRT-PCR) and Western blot (WB); PTEN m⁶A levels were measured by RNA immunoprecipitation-qPCR (RIP-qPCR); myelin ultrastructure, expression of myelin basic protein (MBP) and neurofascin 155 (NF155), and dendritic spine density were observed using transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), immunofluorescence, qRT-PCR, and primary neuron culture. ResultsBehaviorally, knockdown of METTL14 significantly counteracted the beneficial effects of acupuncture in improving self-grooming, open field exploration, three-chamber social interaction, and Morris water maze learning and memory (P<0.05, P<0.01). Compared with the acupuncture+sh-NC group, the acupuncture+sh-METTL14 group showed significantly decreased mRNA and protein expression of hippocampal METTL14 (P<0.01), and the upregulating effects of acupuncture on METTL3 and PTEN expression were reversed (P<0.01). Meanwhile, knockdown of METTL14 significantly inhibited the acupuncture-induced increase in PTEN m⁶A levels (P<0.01). Morphologically, knockdown of METTL14 attenuated the improvement of myelin structure by acupuncture, reversed the downregulation of MBP and upregulation of NF155 induced by acupuncture, and blocked the increase in dendritic spine density (P<0.05, P<0.01). ConclusionMETTL14 is a key molecule mediating the therapeutic effect of acupuncture at Neiguan. Acupuncture at Neiguan upregulates METTL14, thereby enhancing m⁶A methylation modification of PTEN mRNA to stabilize its expression, ultimately promoting myelin development and improving behavioral symptoms in ASD juvenile rats. This preliminarily reveals the modern biological connotation of “opening Xuanfu and dredging myelin”.

Objective:To systematically evaluate the impact of common disinfection methods on key human milk bioactive components [lactoferrin, lysozyme, immunoglobulin A (IgA), lipase, milk fat globule structure] and their sterilization efficacy.Methods:We searched eight databases (CNKI, Wanfang, VIP, Yiigle, PubMed, Web of Science, Cochrane Library, CINAHL) from inception to January 22, 2024, including 25 controlled studies. Network meta-analysis was performed using Stata 17.0, with surface under the cumulative ranking curve (SUCRA) values ranking seven methods: holder pasteurization (HoP), high-temperature short-time pasteurization (HTST), freezing, high-pressure processing (HPP), high hydrostatic pressure (HHP), ultraviolet-C (UV-C), and thermal ultrasound (TUS). Primary outcomes included bioactive component retention rates, milk fat globule integrity, and sterilization efficacy against representative pathogens [ Staphylococcus aureus, cytomegalovirus (CMV)]. Results:Bioactive component retention varied significantly: HPP demonstrated optimal lactoferrin preservation (SUCRA=98.2%, retention rate=92.4%±3.1%); HHP showed superior lysozyme retention (SUCRA=91.0%, retention rate=88.6%±2.5%); HTST maximized IgA preservation (SUCRA=75.5%, retention rate=72.3%±4.2%); UV-C achieved highest lipase retention (SUCRA=92.7%, retention rate=90.1%±1.9%). HPP maintained larger milk fat globules versus HoP. All methods achieved≥6-log pathogen reduction. HPP surpassed HoP/HTST in eliminating S. aureus (6-log vs. 4-log at 5-6 log CFU/ml initial load); HHP reduced CMV from 5.0 to 3.7 log PFU/ml (30 min), outperforming HoP (4.1 log). Non-thermal techniques (HPP/HHP/UV-C) better suppressed pathogen versus HoP. Conclusions:Bioactive component preservation is method-specific clinical protocols should therefore be optimized according to target component: HPP is recommended for lactoferrin/milk fat globule integrity; HHP for lysozyme; HTST for IgA; UV-C for lipase. While all methods meet safety standards, non-thermal technologies offer superior long-term pathogen suppression, providing optimized options for milk banks and clinical breastfeeding support.

Objective To investigate the association of digit ratio with single nucleotide polymorphism(SNP)at three loci(rs17576,rs3918249,rs9509)of matrix metallopeptidase 9(MMP-9)gene.Methods A total of 804 Ningxia Han youths(399 males and 405 females)were used as the study subjects.A digital camera was used to take frontal photographs of the hands,and image analysis software was used to mark the anatomical points and measure the lengths of each finger of both hands(2D,3D,4D,5D);Multiplexed PCR was used to detect the three polymorphic sites of the MMP-9 gene,SPSS 25.0 and R Studio software were used for data analysis and plotting.Results The 2D/3D(P＜0.05)and 2D/4D(left,P＜0.01,right,P＜0.05)of both hands,2D/5D(P＜0.01),3D/5D,4D/5D(P＜0.05)of the right hand,and 3D/4D(P＜0.05)of the left hand in female youths of Ningxia Han were significantly higher than those in males,Differences in genotypes and allele frequencies at all 3 loci of the MMP-9 gene were not statistically significant between genders(P＞0.05).Right hand 2D/4D was significantly associated with genotypes at the rs17576 and rs3918249 loci in male youths(P＜0.05).Conclusion MMP-9 gene SNPs(rs17576 and rs3918249)may be associated with the formation of 2D/4D of Ningxia Han male youths.

Objective:To systematically evaluate the impact of common disinfection methods on key human milk bioactive components [lactoferrin, lysozyme, immunoglobulin A (IgA), lipase, milk fat globule structure] and their sterilization efficacy.Methods:We searched eight databases (CNKI, Wanfang, VIP, Yiigle, PubMed, Web of Science, Cochrane Library, CINAHL) from inception to January 22, 2024, including 25 controlled studies. Network meta-analysis was performed using Stata 17.0, with surface under the cumulative ranking curve (SUCRA) values ranking seven methods: holder pasteurization (HoP), high-temperature short-time pasteurization (HTST), freezing, high-pressure processing (HPP), high hydrostatic pressure (HHP), ultraviolet-C (UV-C), and thermal ultrasound (TUS). Primary outcomes included bioactive component retention rates, milk fat globule integrity, and sterilization efficacy against representative pathogens [ Staphylococcus aureus, cytomegalovirus (CMV)]. Results:Bioactive component retention varied significantly: HPP demonstrated optimal lactoferrin preservation (SUCRA=98.2%, retention rate=92.4%±3.1%); HHP showed superior lysozyme retention (SUCRA=91.0%, retention rate=88.6%±2.5%); HTST maximized IgA preservation (SUCRA=75.5%, retention rate=72.3%±4.2%); UV-C achieved highest lipase retention (SUCRA=92.7%, retention rate=90.1%±1.9%). HPP maintained larger milk fat globules versus HoP. All methods achieved≥6-log pathogen reduction. HPP surpassed HoP/HTST in eliminating S. aureus (6-log vs. 4-log at 5-6 log CFU/ml initial load); HHP reduced CMV from 5.0 to 3.7 log PFU/ml (30 min), outperforming HoP (4.1 log). Non-thermal techniques (HPP/HHP/UV-C) better suppressed pathogen versus HoP. Conclusions:Bioactive component preservation is method-specific clinical protocols should therefore be optimized according to target component: HPP is recommended for lactoferrin/milk fat globule integrity; HHP for lysozyme; HTST for IgA; UV-C for lipase. While all methods meet safety standards, non-thermal technologies offer superior long-term pathogen suppression, providing optimized options for milk banks and clinical breastfeeding support.

Background: Diabetes-induced cardiac fibrosis is one of the main mechanisms of diabetic cardiomyopathy. As a common histone methyltransferase, enhancer of zeste homolog 2 (EZH2) has been implicated in fibrosis progression in multiple organs. However, the mechanism of EZH2 in diabetic myocardial fibrosis has not been clarified. Methods: In the current study, rat and mouse diabetic model were established, the left ventricular function of rat and mouse were evaluated by echocardiography and the fibrosis of rat ventricle was evaluated by Masson staining. Primary rat ventricular fibroblasts were cultured and stimulated with high glucose (HG) in vitro. The expression of histone H3 lysine 27 (H3K27) trimethylation, EZH2, and myocardial fibrosis proteins were assayed. Results: In STZ-induced diabetic ventricular tissues and HG-induced primary ventricular fibroblasts in vitro, H3K27 trimethylation was increased and the phosphorylation of EZH2 was reduced. Inhibition of EZH2 with GSK126 suppressed the activation, differentiation, and migration of cardiac fibroblasts as well as the overexpression of the fibrotic proteins induced by HG. Mechanical study demonstrated that HG reduced phosphorylation of EZH2 on Thr311 by inactivating AMP-activated protein kinase (AMPK), which transcriptionally inhibited peroxisome proliferator-activated receptor γ (PPAR-γ) expression to promote the fibroblasts activation and differentiation. Conclusion Our data revealed an AMPK/EZH2/PPAR-γ signal pathway is involved in HG-induced cardiac fibrosis.

Objective:To analyze the radiographic parameters of shoulder balance that affect the postoperative satisfaction of Lenke type 1 adolescent idiopathic scoliosis (AIS).Methods:A total of 98 patients with AIS who underwent posterior pedicle screw fusion in Xijing Hospital of Air Force Medical University from August 2017 to July 2020 were retrospectively analyzed. There were 26 males and 72 females, aged 15.2±5.3 years (range, 10-24 years). Distribution of upper instrumented vertebrae: T 2 58 cases (59%), T 3 25 cases (26%), T 4 15 cases (15%); Distribution of lower instrumented vertebrae: T 12 63 cases (64%), L 1 28 cases (29%), L 2 4 cases (4%), L 3 3 cases (3%). Clavicle angle (CA), radiographic shoulder height (RSH), and coracoid height difference (CHD), clavicle-rib cage intersection difference (CRID), T 1 tilt angle, first rib tilt angle, clavicle chest angle difference(CCAD) and Scoliosis Research Society-22 (SRS-22) scale were compared before and after operation. Binary logistic regression was used to analyze the radiographic indicators of shoulder balance that affected the postoperative satisfaction of Lenke type 1 AIS. The receiver operating characteristic (ROC) curve was drawn to determine the threshold value of the imaging index. Results:All operations were successfully completed. The operation time was 260±80 min (range, 220-320 min), and the intraoperative blood loss was 360±110 ml (range, 300-700 ml). There was no nerve, dural or vascular injury during operation. RSH, CHD, CRID, T 1 tilt angle, first rib tilt angle, and CCAD at the final follow-up were 4.0 (0, 13.9) mm, 7.0 (0, 12.9) mm, 4.0 (0, 10.0) mm, 4.8° (3.3°, 8.2°), 5.3°±3.9°, and 5.5° (3.0°, 8.9°), respectively, which were less than the preoperative 10.6 (2.0, 20.3) mm, 10.3 (2.5, 15.9) mm, 8.0 (1.0, 15.2) mm, 7.6° (3.5°, 12.2°), 7.5°±6.9°, 8.5° (3.6°, 18.3°), and the difference was statistically significant ( P<0.05). The SRS-22 function, pain, appearance, and psychological scores at the final follow-up were 4.6 (4.0, 4.9), 4.1±0.5, 4.1±0.7, and 4.2 (3.9, 4.8) points, respectively, which were greater than the preoperative scores of 4.2 (3.8, 4.6), 4.0±0.7, 3.5±0.7, and 4.0 (3.5, 4.4) points, the difference was statistically significant ( P<0.05). Binary logistic regression showed that CCAD was an independent radiographic indicator of shoulder balance that affected the satisfaction of AIS patients after orthopaedic surgery ( OR=0.826, P=0.040). ROC curve showed that the area under the curve and 95% CI was 0.726 (0.572, 0.865), and the threshold was 6.6°. Conclusion:CCAD is an independent radiographic parameter of shoulder balance that affects the postoperative satisfaction of AIS. Patients are more likely to achieve a satisfactory outcome when their postoperative CCAD is ≤6.6°, which can be used clinically as a radiographic parameter to assess the efficacy of orthopaedic spine surgery.

Objective:To investigate the current status, trends, and differences among institutions in the application of delivery room continuous positive airway pressure (DRCPAP) for very preterm infants treated in the institutions in the Chinese Neonatal Network (CHNN). Also, to explore the impact of DRCPAP on the outcomes of very preterm infants in China.Methods:A retrospective cohort study was conducted. Based on the CHNN very preterm infant cohort, very preterm infants (gestational ages ranging from 25 weeks +0 to 31 weeks +6) born in-hospital and treated in 79 tertiary neonatal intensive care units (NICUs) participating in the CHNN from 2019 to 2021 were included. The usage rates of DRCPAP in different hospitals, as well as gestational ages and years, were described. Data were analyzed using the Chi-square test (or Fisher's exact test) or t-test. A multivariate logistic regression model was established to explore the correlation between DRCPAP and clinical outcomes. Results:(1) A total of 18 048 very preterm infants were included. Among them, 3 666 (20.3%) received DRCPAP, and 14 382 (79.7%) did not. (2) The usage rate of DRCPAP for very preterm infants among different institutions is from 0.0% to 94.5%. Fourteen institutions did not use DRCPAP, and 55 institutions had a usage rate below 30%. The usage rate of DRCPAP in very preterm infants increased annually, from 13.8% (818/5 916) in 2019 to 26.0% (1 583/6 097) in 2021 ( χ2trend=122.00, P<0.001). (3) The DRCPAP group had higher rates of maternal assisted reproductive technology pregnancy, chorioamnionitis, a full course of antenatal corticosteroids, gestational diabetes, fetal distress, antenatal magnesium sulfate use, and cesarean delivery compared to the non-DRCPAP group [20.3% (744/3 665) vs. 17.6% (2 529/14 369), χ2=14.45; 23.0% (695/3 021) vs. 16.4% (1 956/11 926), χ2=72.57; 57.1% (2 090/3 660) vs. 54.3% (7 766/14 302), χ2=9.55; 23.0% (844/3 669) vs. 20.7% (2 969/14 342), χ2=9.77; 8.7% (319/3 666) vs. 7.0% (1 006/14 371), χ2=12.51; 87.1% (3 186/3 657) vs. 82.0% (11 736/14 312), χ2=81.38; 63.5% (2 327/3 664) vs. 60.7% (8 722/14 369), χ2=9.59; all P<0.05]. While the incidence of hypertensive disorders of pregnancy and the proportion of infants not using antenatal corticosteroids were lower in the non-DRCPAP group [17.1% (626/3 660) vs. 22.6% (3 183/14 084), χ2=44.70; 14.2% (520/3 661) vs. 19.7% (2 814/14 284), χ2=57.34; all P<0.05]. The DRCPAP group had lower birth weight and gestational age, higher 1 min and 5 min Apgar scores, and lower neonatal transport stabilization index scores [(1 308±314) g vs. (1 325±315) g, t=2.90; (29.5±1.7) weeks vs. (29.7±1.6) weeks, t=3.96; (7.9±1.8) scores vs. (7.6±1.9) scores, t=－9.80; (9.0±1.1) scores vs. (8.7±1.3) scores, t=－13.01; (11.0±9.4) scores vs. (13.1±9.8) scores, t=11.31; all P<0.05]. The incidence of early-onset sepsis was higher in the DRCPAP group than in the non-DRCPAP group [1.8% (68/3 578) vs. 1.3% (193/14 296), adjusted OR (95% CI): 1.417 (1.028-1.955)], while the rates of tracheal intubation within 72 hours, PS use, invasive mechanical ventilation, mortality, admission hypothermia, grade Ⅲ-Ⅳ intracranial hemorrhage, and stage≥3 ROP were all lower in the DRCPAP group compared to the non-DRCPAP group [22.6% (830/3 666) vs. 36.9% (5 310/14 374), adjusted OR (95% CI): 0.499 (0.448-0.555); 53.1% (1 947/3 666) vs. 58.3% (8 388/14 377), adjusted OR (95% CI): 0.764 (0.697-0.836); 30.1% (1 104/3 662) vs. 43.9% (6 307/14 366), adjusted OR (95% CI): 0.539 (0.487-0.595); 7.4% (274/3 666) vs. 10.6% (1 526/14 342), adjusted OR (95% CI): 0.709 (0.601-0.836); 57.5% (2 103/3 657) vs. 66.5% (9 501/14 287), adjusted OR (95% CI): 0.722 (0.661-0.788); 3.0% (101/3 366) vs. 5.9% (763/12 932)], adjusted OR (95% CI): 0.525 (0.412-0.669); 2.2% (65/2 954) vs. 3.3% (367/11 121), adjusted OR (95% CI): 0.692 (0.505-0.947); all P<0.05]. There were no statistically significant differences between the two groups in the incidence rates of BPD at a corrected gestational age of 36 weeks, patent ductus arteriosus ligation, spontaneous intestinal perforation, and pneumothorax. Conclusions:Domestically, the application of DRCPAP might be related to a decrease in the demand for invasive ventilation, the use of surfactants, and mortality, but it might not reduce the occurrence of bronchopulmonary dysplasia, at a corrected gestational age of 36 weeks. In recent years, the use of DRCPAP in very premature infants in China has increased, but the overall usage rate is still low, and there are significant differences between units, requiring continuous quality improvement.

Objective:To investigate the current status, trends, and differences among institutions in the application of delivery room continuous positive airway pressure (DRCPAP) for very preterm infants treated in the institutions in the Chinese Neonatal Network (CHNN). Also, to explore the impact of DRCPAP on the outcomes of very preterm infants in China.Methods:A retrospective cohort study was conducted. Based on the CHNN very preterm infant cohort, very preterm infants (gestational ages ranging from 25 weeks +0 to 31 weeks +6) born in-hospital and treated in 79 tertiary neonatal intensive care units (NICUs) participating in the CHNN from 2019 to 2021 were included. The usage rates of DRCPAP in different hospitals, as well as gestational ages and years, were described. Data were analyzed using the Chi-square test (or Fisher's exact test) or t-test. A multivariate logistic regression model was established to explore the correlation between DRCPAP and clinical outcomes. Results:(1) A total of 18 048 very preterm infants were included. Among them, 3 666 (20.3%) received DRCPAP, and 14 382 (79.7%) did not. (2) The usage rate of DRCPAP for very preterm infants among different institutions is from 0.0% to 94.5%. Fourteen institutions did not use DRCPAP, and 55 institutions had a usage rate below 30%. The usage rate of DRCPAP in very preterm infants increased annually, from 13.8% (818/5 916) in 2019 to 26.0% (1 583/6 097) in 2021 ( χ2trend=122.00, P<0.001). (3) The DRCPAP group had higher rates of maternal assisted reproductive technology pregnancy, chorioamnionitis, a full course of antenatal corticosteroids, gestational diabetes, fetal distress, antenatal magnesium sulfate use, and cesarean delivery compared to the non-DRCPAP group [20.3% (744/3 665) vs. 17.6% (2 529/14 369), χ2=14.45; 23.0% (695/3 021) vs. 16.4% (1 956/11 926), χ2=72.57; 57.1% (2 090/3 660) vs. 54.3% (7 766/14 302), χ2=9.55; 23.0% (844/3 669) vs. 20.7% (2 969/14 342), χ2=9.77; 8.7% (319/3 666) vs. 7.0% (1 006/14 371), χ2=12.51; 87.1% (3 186/3 657) vs. 82.0% (11 736/14 312), χ2=81.38; 63.5% (2 327/3 664) vs. 60.7% (8 722/14 369), χ2=9.59; all P<0.05]. While the incidence of hypertensive disorders of pregnancy and the proportion of infants not using antenatal corticosteroids were lower in the non-DRCPAP group [17.1% (626/3 660) vs. 22.6% (3 183/14 084), χ2=44.70; 14.2% (520/3 661) vs. 19.7% (2 814/14 284), χ2=57.34; all P<0.05]. The DRCPAP group had lower birth weight and gestational age, higher 1 min and 5 min Apgar scores, and lower neonatal transport stabilization index scores [(1 308±314) g vs. (1 325±315) g, t=2.90; (29.5±1.7) weeks vs. (29.7±1.6) weeks, t=3.96; (7.9±1.8) scores vs. (7.6±1.9) scores, t=－9.80; (9.0±1.1) scores vs. (8.7±1.3) scores, t=－13.01; (11.0±9.4) scores vs. (13.1±9.8) scores, t=11.31; all P<0.05]. The incidence of early-onset sepsis was higher in the DRCPAP group than in the non-DRCPAP group [1.8% (68/3 578) vs. 1.3% (193/14 296), adjusted OR (95% CI): 1.417 (1.028-1.955)], while the rates of tracheal intubation within 72 hours, PS use, invasive mechanical ventilation, mortality, admission hypothermia, grade Ⅲ-Ⅳ intracranial hemorrhage, and stage≥3 ROP were all lower in the DRCPAP group compared to the non-DRCPAP group [22.6% (830/3 666) vs. 36.9% (5 310/14 374), adjusted OR (95% CI): 0.499 (0.448-0.555); 53.1% (1 947/3 666) vs. 58.3% (8 388/14 377), adjusted OR (95% CI): 0.764 (0.697-0.836); 30.1% (1 104/3 662) vs. 43.9% (6 307/14 366), adjusted OR (95% CI): 0.539 (0.487-0.595); 7.4% (274/3 666) vs. 10.6% (1 526/14 342), adjusted OR (95% CI): 0.709 (0.601-0.836); 57.5% (2 103/3 657) vs. 66.5% (9 501/14 287), adjusted OR (95% CI): 0.722 (0.661-0.788); 3.0% (101/3 366) vs. 5.9% (763/12 932)], adjusted OR (95% CI): 0.525 (0.412-0.669); 2.2% (65/2 954) vs. 3.3% (367/11 121), adjusted OR (95% CI): 0.692 (0.505-0.947); all P<0.05]. There were no statistically significant differences between the two groups in the incidence rates of BPD at a corrected gestational age of 36 weeks, patent ductus arteriosus ligation, spontaneous intestinal perforation, and pneumothorax. Conclusions:Domestically, the application of DRCPAP might be related to a decrease in the demand for invasive ventilation, the use of surfactants, and mortality, but it might not reduce the occurrence of bronchopulmonary dysplasia, at a corrected gestational age of 36 weeks. In recent years, the use of DRCPAP in very premature infants in China has increased, but the overall usage rate is still low, and there are significant differences between units, requiring continuous quality improvement.

OBJECTIVE: To determine the effects of berberine (BBR) on the activation of toll-like receptor 4 (TLR4), nuclear factor (NF)κB (NF-κB) signaling and NLRP3 inflammasome in patients with gout. METHODS: Peripheral blood mononuclear cells (PBMCs) were obtained from 24 acute (AP) and 41 non-acute (NAP) phases of primary gout patients, respectively, as well as 30 healthy controls (HC). TLR4, NF-κB (p65), NLRP3, apoptosis-associated specklike protein containing a CARD (PYCARD), cysteinyl aspartate specific proteinase-1 (CASP1), and interleukin-1β (IL-1β) mRNA expression levels in PBMCs were measured by quantitative reverse transcriptase polymerase chain reaction. The protein levels of TLR4, myeloid differentiation factor 88 (MyD88), NF-κB (p50/65), inhibitor of kappa B kinase α/β (IKKα/β), NF-κB inhibitor α (IKBα), phospho-IKKα/β (p-IKKα/β), NLRP3, PYCARD, and CASP1 were monitored by Western blotting. Serum IL-1β protein level was measured using enzyme-linked immunosorbent assay (ELISA). In addition, PBMCs from HC and macrophages derived from a spontaneously immortalized monocyte-like cell line (THP-1) were stimulated using monosodium urate (MSU, 100 µg/mL), 0.1% dimethyl sulfoxide, 25 µmol/L BBR, and 10, 25, and 50 µmol/L BBR+100 µg/mL MSU for different time periods. The protein levels of IL-1β and IL-18 in cell culture supernatants was measured by ELISA, and the protein expressions of TLR4, MyD88, NF-κB (p50/p65), IKKα/β, I κBβ, p-IKKα/β, NLRP3, PYCARD, and CASP1 in macrophages were analyzed by Western blotting. RESULTS: (1) TLR4, NF-κB (p65), PYCARD, CASP1, and IL-1β mRNA levels in PBMCs were significantly higher in the AP group than in the HC group (P<0.05). The NLRP3 mRNA expression levels in PBMCs were found to be significantly lower in the AP and NAP groups than in the HC group (P<0.05, P<0.01). (2) The protein levels of TLR4, IKKβ, MyD88, NF-κB, p-IKKα/β, PYCARD, and CASP1 in PBMCs were significantly higher, and those of IκBα, IKKα, and NLRP3 were found to be significantly lower in the AP group than in the HC group (P<0.05 or P<0.01). (3) The serum IL-1β protein levels were significantly higher in the AP and NAP groups than in the HC group (P<0.01). (4) The IL-1β protein level was significantly lower in the culture supernatants of the PBMCs stimulated with MSU for 3 and 6 h in the 25 and 50 µmoL/L BBR groups compared with that in the MSU group (P<0.01). (5) The protein levels of IL-1β and IL-18 were also significantly lower in the culture supernatants of macrophages stimulated with MSU for 3 and 6 h in BBR groups compared with those in the MSU group (P<0.01). (6) The protein levels of TLR4, MyD88, NF-κB (p50, p65, p105), IKKα/β, p-IκBα, p-IKKα/β, PYCARD, and CASP1 were significantly differed between the macrophages stimulated with MSU for 0.5 and 6 h in BBR groups compared with those in the MSU group (P<0.05 or P<0.01). CONCLUSIONS Activation of TLR4-NFκB signaling and NLRP3 inflammasome by MSU crystals drives the progression of gout inflammation. BBR ameliorates gouty inflammation, which is mechanistically associated with its regulation of TLR4-NF-κB signaling and NLRP3 inflammasome expression.