Objective To establish the method of fingerprint and content determination of multi-component for Xiangsha Yangwei pill by gas chromatography(GC).Methods The GC fingerprint of Xiangsha Yangwei pill was found,and the peak attribution was carried out.The contents of limonene,eucalyptol,camphor,borneol,bornyl acetate,patchouli alcohol,pogostone,and α-cyperone were determined.Results The fingerprint similarity of 56 batches of Xiangsha Yangwei pill were 0.33-0.99,28 common peaks were confirmed,and 14 known components were identified.Limonene,eucalyptol,camphor,borneol,bornyl acetate,patchouli alcohol,pogostone and α-cyperone showed good linearity within the determined ranges(14.30-286.08,24.52-490.44,16.14-322.88,9.40-187.95,15.39-307.83,25.78-515.60,19.95-398.90,and 24.87-497.30 μg·mL-1).The average recoveries were 101.20％,97.90％,93.97％,94.23％,102.94％,100.54％,99.16％,and 98.31％;with the RSDs were 2.41％,1.48％,1.65％,2.00％,1.93％,2.30％,2.07％,and 2.38％,respectively.The concentrations of eight components were 0.2-959.1,0.3-420.4,1.0-542.6,0.0-64.5,0.0-364.2,0.0-339.6,0.0-130.7,0.0-82.0 μg·g-1,respectively.Conclusion The fingerprint and multi-component determination method can be used for the quality control and evaluation of Xiangsha Yangwei pill.

The fat mass and obesity-associated protein (FTO) is an RNA demethylase required for catalytic demethylation of N ⁶-methyladenosine (m⁶A); it is highly expressed and functions as an oncogene in acute myeloid leukemia (AML). Currently, the overarching objective of targeting FTO is to precisely inhibit the catalytic activity. Meanwhile, whether FTO degradation also exerts antileukemic effects remains unknown. Herein, we designed the first FTO-targeting proteolysis targeting chimera (PROTAC) degrader QP73 using our FTO inhibitor Dac85-which potently inhibits FTO demethylation in AML cell lines-as a warhead. Notably, QP73 significantly induced FTO degradation in a time-, dose-, and ubiquitin-proteasome system-dependent manner and had superior antiproliferative activities to the FTO inhibitor Dac85 in various AML cell lines. Moreover, QP73 treatment significantly increased m⁶A modification on mRNA, promoted myeloid differentiation, and induced apoptosis of AML cells. Quantitative proteomics analysis showed that QP73 induced complete FTO degradation, upregulating RARA and ASB2 abundance and downregulating CEBPA, MYC, PFKP, and LDHB levels in AML cells. Lastly, QP73 exhibited antileukemic activity by increasing m⁶A modification and decreasing FTO levels in xenograft AML tumors. This proof-of-concept study shows that FTO-targeting PROTAC degraders can regulate the FTO signaling pathway and have potential antileukemia applications.

Amyotrophic Lateral Sclerosis (ALS) is a complex neurodegenerative disorder characterized by progressive axonopathy, jointly leading to the dying back of the motor neuron, disrupting both nerve signaling and motor control. In this review, we highlight the roles of axonopathy in ALS progression, driven by the interplay of multiple factors including defective trafficking machinery, protein aggregation, and mitochondrial dysfunction. Dysfunctional intracellular transport, caused by disruptions in microtubules, molecular motors, and adaptors, has been identified as a key contributor to disease progression. Aberrant protein aggregation involving TDP-43, FUS, SOD1, and dipeptide repeat proteins further amplifies neuronal toxicity. Mitochondrial defects lead to ATP depletion, oxidative stress, and Ca²⁺ imbalance, which are regarded as key factors underlying the loss of neuromuscular junctions and axonopathy. Mitigating these defects through interventions including neurotrophic treatments offers therapeutic potential. Collaborative research efforts aim to unravel ALS complexities, opening avenues for holistic interventions that target diverse pathological mechanisms.
Humans ; Amyotrophic Lateral Sclerosis/therapy* ; Animals ; Axons/metabolism* ; Mitochondria/metabolism* ; Motor Neurons/pathology*

Objective To investigate the relationship between promotion of osteogenic differentiation of adipose-derived stem cells (ADSCs) by exosomes secreted by ADSCs and the Wnt/β-catenin signaling pathway. Methods ADSCs were extracted in the Plastic Surgery Department of the First Hospital Affiliated to China Medical University. Exosomes were extracted from ADSCs induced to differentiate into osteoblasts. The ADSCs were separated into four groups and used for measuring activity of alkaline phosphatase (ALP) by using enzyme-labeled assay, formation of calcium nodes by Alizarin red staining, and expression of β-catenin by Western blotting. Results ADSC-derived exosomes promoted differentiation of ADSCs into osteoblasts by upregulating the expression of β-catenin. Addition of Dickkopf-related protein 1 (DKK1), an inhibitor of Wnt/β-catenin pathway, inhibited expression of β-catenin which in turn resulted in reduced potential of ADSCs for osteogenic differentiation. Conclusion The osteogenic differentiation of ADSCs can be promoted by exosomes derived from ADSCs via the Wnt/β-catenin signaling pathway.

Objective: To explore the effect of reducing complication in branched spongiosum repair for hypospadias Duckett surgery. Methods: Retrospective analysis was performed for the clinical data of 140 surgical infants with hypospadias Duckett surgery from January 2016 to April 2018. According to different covering materials on newly formed urethra, the experimental group (68 cases) cut dysplasia, branched spongiosum and lateral Buck′s fascia from tunica albuginea to coronary groove and then sutured over newly formed urethra, and the control group (72 cases) used Dartos fascia or subcutaneous fascia. The operative age, penile curvature, length of urethral defect, operation time, added value of coronary groove before and after operation, maximum urinary flow rate and complication rate of postoperative follow-up were compared between the two groups, and the therapeutic effect was analyzed. Results: There was no statistically significant difference between two groups in operating time[(135±49)min vs.(135±45)min, P=0.580] and the value added of coronary sulcus[(0.1±0.2)cm vs.(0.1±0.1)cm, P=0.167]. In experimental group, there were 8 cases of urethra percutaneous fistula (11.8%) (coronary groove fistula in 3 cases, 4.4%)and 6 cases of urethral stenosis (8.8%) without urethral dehiscence. In control group, there were 18 cases of urethra percutaneous fistula (25.0%)(coronary groove fistula in 11 cases, 15.3%), 10 cases of urethral stenosis (13.9%), and 2 cases of urethral dehiscence (2.8%). The incidence of postoperative urinary fistula and total complications in experimental group was lower than that in the control group, and the difference was statistically significant (P<0.05), while the incidence of urethral stenosis was not statistically different between two groups (P>0.05). The maximum urine flow rate of experimental group at 6 months after surgery was significantly higher than control group[(9.5±3.8)ml/s vs.(6.8±2.8)ml/s], and the difference was statistically significant (P<0.001). Conclusions Urethral cavernosum reconstruction can significantly reduce the incidence of postoperative coronary groove fistula of hypospadias Duckett surgery and improve the postoperative urine flow rate.

Objective To investigate the feasibility of differential expression proteins identification in peripheral blood mononuclear cells (PBMCs) of sepsis patients using Data Independent Acquisition liquid chromatography-mass spectrometry (DIA LC-MS). Methods Prospective studies were employed and targeted at 10 sepsis patients admitted to the Intensive Care Unit (ICU) of Shanghai Fifth People's Hospital Affiliated to Fudan University from April 2016 to July 2016. And 10 patients admitted to the ICU with similar age and sex that were not complicated with sepsis were served as the control group.The proteins from peripheral blood mononuclear cells in 10 sepsis patients and 10 control persons were analyzed using the latest DIA LC-MS technology and the skyline data extraction software; the model of data obtained from the above analysis was further discriminatorily analyzed with principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA) and orthogonal partial least squares discriminant analysis (OPLS-DA); then the differential proteins of peripheral blood mononuclear cells were analyzed by Pathway analysis and GO analysis. The possible markers were identified by preliminary screening according to variable importance in the projection (VIP). The top 3 proteins (VIP > 1, P< 0.05) were verified by ELISA and their ROC curves were analyzed. Results Totally 1062 fragment ions were identified and 119 proteins were obtained. Among them, 31 proteins were up-regulated and 88 proteins down-regulated. Pathway analysis showed that carbon metabolism, platelet activation, bacterial invasion of epithelium, and complement coagulation cascade activation were participated in the development of sepsis. ELISA showed that significant difference of HMGB-1, MMP-8, and LCN2 in the expression of peripheral blood mononuclear cells in the sepsis and control people (P<0.01). The area under the ROC curve is greater than 0.85, which has good sensitivity and specificity. Conclusions DIA-MS is a compelling way for detecting differential expression proteins. PCA, PLSDA and OPLSDA are suitable for pattern recognition. The high expression of HMGB-1, NGAL and MMP-8 in immune cells may be the potential biomarker of the disease, which lays the foundation for research of early diagnosis and treatment of sepsis.