ObjectiveTo explore the potential mechanism underlying electroacupuncture therapy with the midnight-noon ebb-low method with hour-prescription for chronic obstructive pulmonary disease （COPD） with lung-qi deficiency syndrome from the perspective of brain and muscle basic helix-loop-helix ARNT-like 1 （BMAL1） as well as neutrophil extracellular traps （NETs）. MethodsThe experiment was conducted in two phases. For the therapeutic effectiveness observation， 24 rats were randomly allocated into control group 1， model group 1， midnight-noon electro-acupuncture group 1 and conventional electro-acupuncture group， with 6 rats per group. Rat models of COPD with lung-qi deficiency syndrome were established via cigarette smoke exposure combined with intratracheal instillation of lipopolysaccharide （LPS）. After successful modelling， rats in midnight-noon electro-acupuncture group 1 received electro-acupuncture at acupoints "Taiyuan （LU 9）"， "Feishu （BL 13）" and "Zusanli （ST 36）" during 05：00—07：00， 30 minutes per treatment once every other day for 14 consecutive days （7 sessions in total）. Rats in the conventional electro-acupuncture group received identical electroacupuncture manipulation at random daytime hours （08：00—18：00）. Pulmonary function parameters including forced expiratory volume in 0.3 second （FEV_0.3）， forced vital capacity （FVC） and FEV_0.3/FVC ratio were detected. Pulmonary histopathological changes were observed via hematoxylin-eosin （HE） staining. Plasma levels of pro-inflammatory factors interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α）， as well as pulmonary reactive oxygen species （ROS） content， BMAL1 protein expression and the levels of neutrophil extracellular traps （NETs） biomarkers such as myeloperoxidase （MPO）， neutrophil elastase （NE）， and citrullinated histone H3 （CitH3） in lung tissue were determined. For the mechanistic verification phase， 36 rats were divided into control group 2， model group 2， midnight-noon electro-acupuncture group 2， overexpression empty group， overexpression BMAL1 group and overexpression with midnight-noon electro-acupuncture group， with 6 rats in each group. At the 5th week of model construction， rats in the overexpression BMAL1 group and overexpression with midnight-noon electro-acupuncture group received intratracheal instillation of 100 μl adenoviral suspension carrying overexpressed Bmal1 gene. Rats in the overexpression empty group were injected with an equal titer and equal volume of blank adenovirus without the Bmal1 coding sequence into the lung at the identical time point. Midnight-noon electro-acupuncture group 2 and overexpression with midnight-noon electro-acupuncture group were consistent with the aforementioned protocol. Finally， pulmonary levels of MPO， NE and CitH3 were measured in all groups. ResultsCompared with the control group 1， the model group 1 exhibited decreased FEV_0.3， FVC and FEV_0.3/FVC ratio， elevated plasma IL-1β， TNF-α and pulmonary ROS levels， downregulated lung BMAL1 protein expression， and increased contents of MPO， NE and CitH3 （all P＜0.05）； typical NETs-like structures with sparse granular substances attached to fibrous networks were observed under scanning electron microscopy； HE staining revealed damaged alveolar architecture accompanied by inflammatory cell infiltration. Compared with model group 1， both the midnight-noon electro-acupuncture group 1 and conventional electro-acupuncture group achieved obvious improvements in all above indicators， with superior therapeutic effects in midnight-noon electro-acupuncture group 1 （P＜0.05）. Consistently， midnight-noon electro-acupuncture group 1 showed more prominent alleviation of NETs-like structure formation and lung pathological injury compared to the conventional electro-acupuncture group. Mechanistic validation results demonstrated that compared with model group 2， the average optical density of MPO as well as the protein levels of MPO， NE and CitH3 were markedly reduced in midnight-noon electro-acupuncture group 2， overexpression BMAL1 group and overexpression with midnight-noon electro-acupuncture group （P＜0.05）； furthermore， these parameters were significantly lower in the overexpression with midnight-noon electro-acupuncture group than in midnight-noon electro-acupuncture group 2 （P＜0.05）. ConclusionElectro-acupuncture based on the midnight-noon ebb-low method with hour-prescription may alleviate pulmonary inflammation in COPD by upregulating the expression of clock protein BMAL1 and inhibiting excessive accumulation of NETs in lung tissue.

Background: Adrenocortical hormones, particularly 11-oxygenated androgens, are pivotal in female reproductive health and fertility. Standardized detection kits and population-specific reference intervals are lacking in China, hindering related clinical applications. Methods: A HPLC-tandem mass spectrometry (HPLC-MS/MS) pipeline was developed, rigorously validated, and applied to simultaneously quantify corticosterone, cortisone, cortisol, 18-OH cortisol, androstenedione (A4), 11β-hydroxyandrostenedione (11-OH A4), dehydroepiandrosterone, and dehydroepiandrosterone sulfate in serum samples from 455 reproductive-aged women (18–45 yrs) in Guangxi, China. Age-dependent concentration trends were analyzed, and reference intervals stratified by age (2.5th to 97.5th percentiles) were established. Correlations with body-composition metrics, ethnicity, and the menstrual cycle were investigated. Results: The HPLC-MS/MS method demonstrated high precision (intra- and inter-assay CVs < 15%), accuracy, and sensitivity. All eight hormones exhibited significant age-related declines (P < 0.001 for seven hormones; P = 0.001 for 11-OH A4). Notably, 11-OH A4 levels were significantly lower in the 35–45-yr (3.05 nmol/L) and 25–34-yr (3.09 nmol/L) age groups than in the 18–24-yr (3.57 nmol/L) age group, whereas no significant difference was observed between the 35–45-yr and 25–34-yr age groups. Weak negative correlations were observed between the body mass index and corticosterone and cortisone levels, whereas ethnicity and the menstrual cycle showed no significant associations with hormone levels. Conclusions We developed an HPLC-MS/MS-based method for simultaneously quantifying eight adrenocortical hormones, including 11-OH A4, and defined age-specific reference intervals for reproductive-aged Chinese women. These findings advance the clinical utility of adrenocortical hormones in diagnosing and managing reproductive disorders.

[Objective] To establish a cryopreservation protocol for small-volume (≤1 mL) red blood cells (RBCs) and to evaluate the reactivity and stability of blood group antigens after cryopreservation, so as to explore its potential application in immunohematology reference laboratories. [Methods] Small-volume RBCs were cryopreserved for 120 days, followed by thawing and deglycerolization to restore the RBC components. The quality of the RBCs was assessed. Serum antibodies were serially diluted and reacted with RBCs before and after cryopreservation, and agglutination scores were recorded to quantitatively evaluate the reactivity and stability of blood group antigens such as Rh, Duffy, Lewis, Kidd, M, and H. Flow cytometry was used to analyze the percentage and mean fluorescence intensity of ABO antigen expression on RBCs before and after cryopreservation to assess the usability of cryopreserved RBCs in flow immunophenotyping and blood group subtype studies. [Results] The hemolysis rate of thawed and deglycerolized RBCs was (0.27±0.10)%, with a supernatant free hemoglobin level of (0.52±0.14) g/L, and the RBC recovery rate was (69.12±7.91)%. The direct antiglobulin test (DAT) was negative for all thawed RBCs. There was no difference in the reactivity of blood group antigens before and after cryopreservation, and no difference in the percentage and mean fluorescence intensity of A and B antigen expression on RBCs before and after cryopreservation. [Conclusion] The small-volume RBC cryopreservation protocol can be applied to immunohematology analysis in reference laboratories and is expected to be widely used in blood group identification, antibody screening, identification, and blood group-related research.

Objective:To explore the diagnostic value of transcranial magnetic stimulation (TMS), transsacral magnetic root stimulation combined with sacral reflexes, external anal sphincter electromyography and pudendal nerve somatosensory evoked potentials in the assessment of neurogenic lower urinary tract dysfunction (NLUTD).Methods:Twenty-one NLUTD patients (1 with a supra-pontine lesion, 5 with a spinal cord injury, 5 with a cauda equina injury, and 10 with pelvic floor disorders) were enrolled. Needle electromyography (EMG) was used to record TMS-induced and transsacral magnetic stimulation-induced motor evoked potentials (tc-MEPs and ts-MEPs, respectively) related to the external anal sphincter (EAS). The dorsal nerve of the penis or clitoris was stimulated electrically to record the latency of the sacral reflex related to the EAS. Central motor conduction time (CMCT) and the tc/ts-MEP latency ratio were calculated to distinguish central from peripheral lesions.Results:In the one patient with a supra-pontine lesion, although the tc-MEP and ts-MEP latencies were within normal limits, the CMCT was prolonged (28.2ms) and the tc/ts-MEP ratio was large (7.4). Among the five patients with a spinal cord injury, one exhibited prolonged tc-MEP latency (50.6ms) and CMCT (47.8ms), along with a large tc/ts-MEP ratio (18.1). In the five patients with cauda equina injury and the ten with NLUTD secondary to pelvic floor disorders, CMCT was within the normal range [averaging (22.9±4.9ms) and (24.2±3.5ms), respectively], but the ts-MEP latency was prolonged [(7.1±2.1ms) and (8.6±3.7ms), respectively], and the tc/ts-MEP ratio was small [(4.4±0.9) and (4.3±1.5), respectively]. The tc/ts-MEP ratio demonstrated the best rate of abnormality detection (93.8%), with an area under the curve of 0.99, indicating good sensitivity.Conclusions:The tc/ts-MEP ratio can be useful for distinguishing central and peripheral lesions. A markedly increased tc/ts-MEP ratio may suggest central nervous system injury, whereas a decreased ratio may indicate peripheral nervous system injury.

This study was a single-center retrospective study, to investigate the correlation between the relative level of miR-200a in exfoliated cells of peritoneal dialysis (PD) effluent and clinical indicators of peritoneal function in PD patients. Clinical data of PD patients who underwent treatment in the Department of Nephrology, the First Affiliated Hospital of Nanchang University, from January 2024 to December 2024 were collected, and the relative level of miR-200a in exfoliated cells of PD effluent was assessed. Patients were divided into three groups according to dialysis duration (Group A:≤1 year; Group B:>1 to <5 years; Group C:≥5 years), and differences in clinical data among the groups were compared. The correlation between the relative level of miR-200a and serum albumin, serum calcium, serum phosphorus, serum creatinine, hemoglobin, the neutrophil/high-density lipoprotein cholesterol ratio, the monocyte/high-density lipoprotein cholesterol ratio, the lymphocyte/high-density lipoprotein cholesterol ratio, and 24-hour urine volume in PD patients was analyzed. The results showed that there were no statistically significant differences among the three groups in gender, age, creatinine clearance rate, or Kt/V (all P>0.05). However, statistically significant differences were found among the three groups in peritoneal transport type ( χ2=13.518, P=0.001) and ultrafiltration volume ( H=6.905, P=0.032). Based on the 4-hour PD effluent creatinine/blood creatinine in the peritoneal equilibration test, patients were divided into the low transporter/low average transporter group (L group) and the high transporter/high average transporter group (H group). A statistically significant difference in the relative level of miR-200a was observed between the two groups [11.70 (1.09, 20.49) vs. 0.73 (0.46, 1.98), t=4.545, P<0.001]. The relative level of miR-200a was positively correlated with ultrafiltration volume and 24-hour urine output ( r=0.328, P=0.030; r=0.516, P<0.001), and negatively correlated with dialysis vintage ( r=-0.496, P<0.001). These results indicate that miR-200a level in PD effluent may, to some extent, reflect patients' peritoneal membrane function.

Various metabolic enzymes and signaling molecules in the reprogramming process of glucose metabolism are involved in the occurrence and development of lung cancer. The study of these metabolic enzymes and signaling molecules is one of the hot spots and directions in the clinical diagnosis and treatment of lung cancer. Fructose-bisphosphate aldolase A (ALDOA) is an important catalytic enzyme in the reprogramming of glucose metabolism, and the abnormal expression of ALDOA is intricately related to the occurrence and development of lung cancer. Systematically exploring of the role of ALDOA in lung cancer metabolism may provide new ideas for predicting the metastasis, prognosis, and treatment after drug resistance of lung cancer.

Objective Fasting hypometabolism regulation technology has broad application potential in long-term space flight and survival in extreme extraterrestrial environments.In-depth research on the substrate conversion of energy metabolism and the formation of new steady states under fasting hypometabolism will provide theoretical basis and experimental data support for formulating effective prolonged fasting application mode.Methods 30 SD rats were randomly divided into control group and fasting group(fasting for 1,2,3,and 5 days).Blood biochemical examination,qRT-PCR,and western blotting were performed to analyze the body weight,blood biochemistry,and expression changes of genes and proteins related to glucose and lipid metabolism during different fasting periods.Results Prolonged fasting significantly reduced the body weight,blood glucose,and triglyceride levels of rats;increased the blood ketone level,and replaced glucose as the main energy substance in the body.There are temporal and tissue-specific changes as a whole.Hepatic and renal gluconeogenesis play major roles respectively during different fasting periods.As the fasting time prolongs,the level of hepatic gluconeogenesis gradually decreases,the content of FFA in the blood increases,the expression level of genes related to fat synthesis decreases,fatty acid oxidation is enhanced,and the expression level of the key gene HMGCS2 for ketone body generation increases.Conclusion During prolonged fasting,there is a significant conversion of glucose-ketone energy supply substrates,and a new steady state of energy metabolism mainly supplied by ketone bodies is formed within 2-5 days of fasting.The body maintains a low metabolic state by regulating changes in key genes in pathways such as glucose and lipid metabolism.

To analyze the characteristics of outpatient rare disease admissions at Xi′an Children′s Hospital based on the two batches of the China′s Rare Disease Catalogs.

Objective:To investigate the role and mechanism of TrxR1 in reprogramming tumor-associated macrophage in breast cancer,providing novel insights and theoretical foundations for clinical breast cancer treatment.Methods:TISIDB database was used to analyze the relationship between TXNRD1(encoding TrxR1)and tumor immunity.Mouse breast cancer 4T1 cells conditioned medium was collected and co-cultured with bone marrow-derived macrophage(BMDM)cells for 48 h to detect the expression of macrophage immunosuppression-related factors.TrxR1 secretion by tumor cells was measured using ELISA kits.TXNRD1 knockdown efficiency was verified via Western blot.Fluorescence quantitative PCR(qPCR)and flow cytometry were used to detect the expression levels of macrophage immunosuppressive factors after TXNRD1 knockdown in tumor cells.JASPAR database was used to analyze the potential regulatory factors,and Western blot was used to verify the expression of pathway-related proteins.Results:Database analysis found that TXNRD1 expression positively correlated with survival risk indices across multiple cancers,with the strongest association observed in breast cancer.Further analysis found that elevated TXNRD1 expression correlated with reduced infiltration of M1 macrophages and natural killer(NK)cells,but increased M2 macrophage infiltration.qPCR and flow cytometry demonstrated that tumor-conditioned medium enhanced macrophage immunosuppression,whereas medium from TXNRD1-knockdown tumor cells suppressed this effect.And TrxR1-neutralizing antibodies could also reversed this effect.JASPAR database analysis identified STAT3 and STAT6 as potential transcriptional regulators,and Western blot confirmed that TXNRD1-knockdown tumor cells conditioned medium inhibited STAT6 pathway activation in macrophages.Conclusion:In the tumor microenvironment,breast tumor-derived TrxR1 promotes macrophage immunosuppression,potentially through activation of the STAT6 signaling pathway.

Objective:To elucidate the regulatory role of matrix metallopeptidase 14 (MMP14) in the migration of chondrogenic precursor cells, thereby providing data support for investigating the pathogenesis of microtia.Methods:An in vitro differentiation model was established using human induced pluripotent stem cells (iPSCs) sequentially induced into neural crest cells (iNCCs) and subsequently into chondrogenic precursor cells (iCPCs), combined with lentivirus-mediated knockdown of MMP14, to investigate the effects of MMP14 on the biological characteristics of iCPCs, including proliferation, differentiation, and migration. Collective cell migration was assessed using scratch wound healing and Transwell migration assays; directional migration was characterized via high-content live-cell imaging; single-cell adhesion force was measured using a micromanipulation system. Collagen degradation was evaluated through hydroxyproline digestion assays. Cell proliferation was analyzed using the CCK-8 assay, and the expression of osteogenic/chondrogenic-related genes (SOX5/6/9, COL1A1, COL2A1, RUNX2, TWIST1) were quantified by real-time quantitative PCR. Immunofluorescence staining was used to assess the expression of F-actin and CD44 proteins. Additionally, transcriptomic sequencing was performed on iCPCs before and after MMP14 knockdown. Results:iPSC→iNCC→iCPC differentiation model was established in vitro. The resulting iCPCs expressed osteo/chondrogenic marker genes, including SOX5, SOX6, SOX9, COL1A1, COL2A1, RUNX2, and TWIST1, and exhibited positive expression of mesenchymal stem cell markers CD90, CD105, and CD73. Upon further induction, functional cartilage spheroids were formed. Compared with normal auricular chondrocytes, auricular chondrocytes from microtia patients showed reduced expression of MMP14 at both mRNA and protein levels. Lentivirus-mediated shRNA knockdown of MMP14 in iCPCs resulted in a marked decrease in its mRNA and protein expression. MMP14 knockdown significantly impaired collective migration of iCPCs, as evidenced by reduced wound closure rates in scratch assays and decreased numbers of migrated cells in Transwell assays. High-content live-cell imaging revealed that MMP14-deficient iCPCs displayed more erratic migration trajectories and a lower straight-line migration ratio. Single-cell adhesion assays showed extracellular matrix (ECM)-dependent alterations: cell adhesion was enhanced on matrigel-coated surfaces but weakened under uncoated conditions. MMP14 knockdown also led to reduced proliferation, decreased collagen degradation, diminished F-actin expression, fewer peripheral adhesion sites, and downregulation of CD44 protein expression, without significantly affecting the expression of chondrogenic genes such as SOX6, SOX9, COL1A1, COL2A1, RUNX2, and TWIST1. Transcriptomic analysis further revealed that MMP14 knockdown significantly downregulated genes involved in extracellular matrix organization, cell adhesion, migration, and tissue development, with enrichment in pathways including ECM-receptor interaction, focal adhesion, and MAPK signaling. Conclusion:MMP14 plays a critical role in the directional migration of chondrogenic precursor cells by regulating ECM remodeling, adhesion signaling, and cytoskeletal proteins.