To explore the chemical constituents of Ecballium elaterium and their cytotoxicity, this study employed multiple chromatographic techniques including normal-phase silica gel, MCI, octadecylsilyl(ODS), Sephadex LH-20 gel, and semi-preparative liquid chromatography for compound isolation from its active fraction. A total of 12 compounds were obtained, and they were identified according to the analysis of a variety of spectral data and literature comparison as 24Z-20,27-dihydroxy-16α,23α-epoxy-cucurbita-2-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranoside(1), cucurbitacin R(2), cucurbitacin B(3), cucurbitacin D(4), cucurbitacin I(5), cucurbitacin L(6), dehydrodiconiferyl alcohol(7), 3-hydroxy-4-methoxycinnamic acid(8), ferulaic acid(9), p-coumaric acid(10), rutin(11), and lariciresinol-4'-O-β-D-glucoside(12), among which compound 1 was a new compound. Compounds 2-6 had strong cytotoxicity against human lung carcinoma A549 cells with the IC_(50) values of(0.48±0.09),(0.03±0.002),(0.13±0.03),(0.87±0.14),(0.15±0.03) μmol·L~(-1), respectively, which were stronger than the positive control doxorubicin \[IC_(50)=(3.92±1.60) μmol·L~(-1)\].
Humans ; Drugs, Chinese Herbal/toxicity* ; Cell Line, Tumor ; Cucurbitaceae/chemistry* ; Cell Survival/drug effects*

Cyclin-dependent kinase 9 (CDK9) is a member of the transcription CDK subfamily and plays a role in transcriptional regulation. Selective CDK9 degraders possess potent clinical advantages over reversible CDK9 inhibitors. Herein, we report the first ATG101-recruiting selective CDK9 degrader, AZ-9, based on the hydrophobic tag kinesin degradation technology. AZ-9 showed significant degradation effects and selectivity toward other homologous cell cycle CDKs in vitro and in vivo, which could also affect downstream related phenotypes. Mechanism research revealed that AZ-9 recruits ATG101 to initiate the autophagy-lysosome pathway, and forms autophagosomes through the recruitment of LC3, which then fuses with lysosomes to degrade CDK9 and the partner protein Cyclin T1. These dates validated the existence of non-proteasomal degradation pathway of hydrophobic driven protein degradation strategy for the first time, which might provide research ideas for chemical induction intervention on other types of pathogenic proteins.

OBJECTIVE From the perspective of China’s health system， to evaluate the cost-effectiveness of maintenance therapy with clopidogrel or aspirin monotherapy in percutaneous coronary intervention （PCI） patients after dual antiplatelet therapy （DAPT）. METHODS A Markov model was adopted with a research period of 25 years and a cycle period of 1 year. Cohort simulations were conducted respectively for the clopidogrel group and aspirin group to predict and compare the long-term economic and health outcomes of PCI patients receiving either clopidogrel or aspirin monotherapy maintenance regimens after DAPT， and cost-effectiveness analysis was conducted. The willingness-to-pay （WTP） threshold was set at the level of 1 times China’s per capita gross domestic product （GDP） in 2024[95 749 yuan per quality-adjusted life year （QALY）]， and the incremental cost- effectiveness ratio （ICER） was calculated. The robustness of the basic analysis results was verified by using single-factor sensitivity analysis and probabilistic sensitivity analysis. RESULTS After PCI， patients received DAPT， clopidogrel monotherapy maintenance treatment reduced the occurrence of death events， and aspirin monotherapy maintenance treatment had a lower probability of stroke and myocardial infarction events. The ICER of the clopidogrel group regimen compared with the aspirin group regimen was 34 644.87 yuan/QALY， which was less than the WTP threshold set in this study. The results of univariate sensitivity analysis indicated that notable uncertainties affecting the basic analysis results were the probability of event-free progression to death in the aspirin group， the event-free cost in the clopidogrel group， and the event-free cost in the aspirin group. The results of probabilistic sensitivity analysis indicated that when the WTP threshold was 95 749 yuan /QALY， the economic probabilities of the clopidogrel group and the aspirin group were 83% and 17%， respectively. The economic probability of the clopidogrel group regimens showed an upward trend with the increase of the WTP threshold. CONCLUSIONS Compared to aspirin monotherapy for maintenance therapy， under the WTP threshold of 1 times China’s per capita GDP in 2024， receiving clopidogrel monotherapy for maintenance therapy after DAPT in PCI patients is more cost-effective.

The mandibular condyle is a critical growth center in craniofacial bone development, especially during postnatal stages. Postnatal condyle osteogenesis requires precise spatiotemporal coordination of growth factor signaling cascades and hierarchical gene regulatory networks. Plagl1, which encodes a zinc finger transcription factor, is a paternally expressed gene. We demonstrate that PLAGL1 is highly expressed in cranial neural crest cell (CNCC)-derived lineage cells in mouse condyles. Using the CNCC-derived lineage-specific Plagl1 knockout mouse model, we evaluate the function of PLAGL1 during postnatal mouse condyle development. Our findings show that PLAGL1 contributes significantly to osteoblast differentiation, and its deficiency impairs osteogenic lineage differentiation, which consequently disrupts mandibular condyle development. Mechanistically, insulin-like growth factor 2 (IGF2) in complex with IGF-binding proteins (IGFBPs) has been identified as the principal PLAGL1 effector responsible for osteogenic regulation during postnatal condyle morphogenesis. Plagl1 deficiency significantly downregulates the IGF2/IGFBP pathway, leading to disordered glucose metabolism, defective extracellular matrix organization, and impaired ossification. Exogenous IGF2 treatment rescues impaired osteoblast differentiation caused by Plagl1 deficiency. In conclusion, the PLAGL1-IGF2 axis is a critical regulator of osteogenesis during mandibular condyle development.
Animals ; Osteogenesis/genetics* ; Insulin-Like Growth Factor II/metabolism* ; Mice ; Transcription Factors/metabolism* ; Mice, Knockout ; Cell Differentiation ; DNA-Binding Proteins/genetics* ; Mandibular Condyle/growth & development* ; Osteoblasts/cytology* ; Signal Transduction ; Neural Crest/cytology*

Objective To explore the effect of long non-coding RNA(long non-coding RNA,lncRNA)small RNA host gene 15(small nucleolar RNA host gene 15,SNHG 15)regulating microRNA 95-3p(miR-95-3p)on breast cancer cell proliferation and apoptosis.Methods Breast cancer cells were divided into control group,knockdown group,interference group and co-transfection group.Human breast cancer cell line MDA-MB-231 was cultured in vitro.Measure the relative expression levels of SNHG 15 mRNA and miR-95-3p mRNA;Methylthiazolyldiphenyl-tetrazolium bromide(MTT)was used to detect the proliferation ability of breast cancer cells;The apoptosis of breast cancer cells was detected by flow cytometry;The migration ability of breast cancer cells was detected by scratch test;Transwell invasion test was used to measure the number of breast cancer cells invaded;Western blot was used to determine the relative expression levels of caspase-3,B-cell lymphoma-2(Bcl-2),and Bcl-2 associated X protein(Bax).Results Compared with control group,the relative expression level of SNHG 15 mRNA and miR-95-3p mRNA,proliferation ability,proliferation rate,migration ability,number of invasion,Bcl-2 protein expression were decreased,but the apoptosis status,apoptosis rate,caspase-3 and Bax expression were increased in knockdown group(P＜0.05).Compared with knockdown group and interference group,the relative expression level of SNHG 15 mRNA and miR-95-3p mRNA,proliferation capacity,proliferation rate,migration capacity,number of invasion,Bcl-2 protein expression were increased,but apoptosis status,apoptosis rate,caspase-3 and Bax expression were decreased in co-transfection group(P＜0.05).Compared with interference group,the relative expression level of SNHG 15 mRNA and miR-95-3p mRNA,proliferation ability,proliferation rate,migration ability,number of invasion,Bcl-2 protein expression were increased,but apoptosis status,apoptosis rate,caspase-3 and Bax expression were decreased in co-transfection group(P＜0.05).Conclusion lncRNA SNHG 15 can affect the proliferation and apoptosis of breast cancer cells by regulating miR-95-3p expression,and is positively associated with the occurrence and development of breast cancer cells.

Objective:To evaluate the clinical efficacy of pushing reduction with our self-designed spinal fracture reduction device in the treatment of A3N0/1 thoracolumbar fractures.Methods:A retrospective study was conducted to analyze the medical records of 53 patients who had undergone surgery for thoracolumbar vertebrae fracture at Department of Minimally Invasive Spine Surgery, Zhengzhou Orthopedic Hospital from January 2019 to January 2022. All patients were treated by internal fixation via the Wiltse approach and bone grafting through the pedicle of the injured vertebrae. Clinical data: 35 males and 18 females; age: (37.8±10.2) years; injured segments: 23 cases at the thoracic spine and 30 cases at the lumbar spine; time from injury to surgery: (3.3±1.5) days. According to whether our self-designed spinal fracture reduction device was used or not, the patients were assigned into group A (23 cases) in which the injured vertebrae were pushed and reduced using our novel spinal fracture reduction device after vertebral distraction reduction by the pedicle screw and group B (30 cases) in which the injured vertebrae were distracted and reduced using the pedicle screw alone. The operation time, intraoperative blood loss and complications were compared between the 2 groups. The anterior vertebral body height ratio (AVBHr), middle vertebral body height ratio (MVBHr), posterior vertebral body height ratio (PVBHr), Cobb angle of the injured vertebra, visual analogue scale (VAS) and Oswestry disability index (ODI) at preoperation, postoperative 3 and 6 months, and the last follow-up were compared between the 2 groups.Results:There was no statistically significant difference in the preoperative general data between the 2 groups, indicating comparability ( P>0.05). All patients were followed up for (16.3±5.9) months. All incisions healed at one stage postoperatively without any related complications. The operation time in group A was significantly longer than that in group B [(115.1±16.6) min. versus (101.0±11.5) min.], the intraoperative blood loss in group A was significantly greater than that in group B [(136.5±17.0) mL versus (121.6±19.8) mL], the MVBHr at postoperative 3 months in group A (93.9%±4.0%) was significantly better than that in group B (83.3%±7.6%), and the MVBHr, AVBHr, Cobb angle, VAS, and ODI at the last follow-up in group A [86.6%±5.5%, 89.8%±4.1%, 4°(4°, 6°), 1 (0, 1) point, and 4.7%±2.0%] were significantly better than those in group B [78.0% (74.0%, 79.0%), 84.5%±4.9%, 12.2°±3.3°, 2 (1, 3) points, and 7.3%±2.7%] (all P<0.05). However, there was no statistically significant difference in PVBHr between the 2 groups at postoperative 3 months or at the last follow-up ( P>0.05). Conclusion:In the treatment of A3N0/1 thoracolumbar fractures, pushing reduction with our self-designed spinal fracture reduction device can directly and effectively reduce the fracture zone of the injured vertebra, which is conducive to maintaining postoperative vertebral reduction, reducing vertebral height loss and kyphotic deformity at a later stage, relieving lumbar pain and improving lumbar spine function.

Ten compounds were separated by various modern chromatographic methods from mastic. They were identified by HR-ESI-MS, IR, UV, NMR, quantum chemical calculation of NMR(qcc-NMR) and comparison with reported data in literature as 17β-hydroxy-28-norolean-18-en-3-one(1), 28-norolean-12,17-dien-3,11-dione(2), 28-norolean-16,18-dien-3-one(3),(24Z)-26-hydroxy-7,24-dientirucalla-3-one(4), masticadienonic acid(5)、masticadienolic acid(6)、erythrodiol(7), 3β,28-dihydroxyoleana-11,13(18)-diene(8), 3-oxo-olean-9(11),12-dien-28-oic acid(9), and 12-oleane-3,11-dione(10). Among them, compound 1 was a novel compound and compound 2 was a novel natural product. Compounds 1-2, 4, and 7-9 were isolated from mastic for the first time. Compound 1 significantly inhibited lipopolysaccharide-induced production of nitric oxide in mouse monocyte macrophage RAW264.7 cells with the IC_(50) of(7.44±0.49) μmol·L~(-1), which was more potent than the positive control, dexamethasone \[IC_(50)=(9.93±1.17) μmol·L~(-1)\]. Compounds 2-3, 5, and 9-10 also showed inhibitory effects, with the IC_(50) ranging from 14.82 to 28.82 μmol·L~(-1). Compounds 4-5, and 7 markedly inhibited the growth of human colon adenocarcinoma cells SW480, with the IC_(50) of(16.13±1.69),(27.40±1.81), and(10.01±0.10) μmol·L~(-1), respectively.
Mice ; Animals ; Mastic Resin/chemistry* ; Humans ; Macrophages/metabolism* ; Triterpenes/isolation & purification* ; Nitric Oxide ; Molecular Structure ; RAW 264.7 Cells ; Drugs, Chinese Herbal/isolation & purification*

Abstract Female stress urinary incontinence (SUI) is a major global public health problem, and its involuntary urinary leakage may seriously affect patients' quality of life. The pathophysiological changes in muscle, connective and nerve tissues induced by diabetes have been found to be strongly correlated with the development and progression of SUI. This review summarizes the association between diabetes and SUI in women and the underlying pathophysiological mechanisms, so as to provide insights into prevention of SUI among female patients with diabetes.

Ketone bodies have beneficial metabolic activities, and the induction of plasma ketone bodies is a health promotion strategy. Dietary supplementation of sodium butyrate (SB) is an effective approach in the induction of plasma ketone bodies. However, the cellular and molecular mechanisms are unknown. In this study, SB was found to enhance the catalytic activity of 3-hydroxy-3-methylglutaryl-CoA synthase 2 (HMGCS2), a rate-limiting enzyme in ketogenesis, to promote ketone body production in hepatocytes. SB administrated by gavage or intraperitoneal injection significantly induced blood ß-hydroxybutyrate (BHB) in mice. BHB production was induced in the primary hepatocytes by SB. Protein succinylation was altered by SB in the liver tissues with down-regulation in 58 proteins and up-regulation in 26 proteins in the proteomics analysis. However, the alteration was mostly observed in mitochondrial proteins with 41% down- and 65% up-regulation, respectively. Succinylation status of HMGCS2 protein was altered by a reduction at two sites (K221 and K358) without a change in the protein level. The SB effect was significantly reduced by a SIRT5 inhibitor and in Sirt5-KO mice. The data suggests that SB activated HMGCS2 through SIRT5-mediated desuccinylation for ketone body production by the liver. The effect was not associated with an elevation in NAD⁺/NADH ratio according to our metabolomics analysis. The data provide a novel molecular mechanism for SB activity in the induction of ketone body production.
Mice ; Animals ; Butyric Acid/metabolism* ; Ketone Bodies/metabolism* ; Liver/metabolism* ; Hydroxybutyrates/metabolism* ; Down-Regulation ; Sirtuins/metabolism* ; Hydroxymethylglutaryl-CoA Synthase/metabolism*

Objective:To investigate the effects of PDCA method on improving the accuracy of the Autar Deep Venous Thrombosis (DVT) Scale used by nurses to assess the risk of deep venous thrombosis in surgical patients.Methods:Applied the way of PDCA, namely, raising questions, analyzing reasons, implementing measures, feeding back effects to professional explanate the difficulty in using the scale, moreover, training the nurses used level education method between August 2017 and December2018.The accuracy and consistency of the scale were compared before and after interventions.Results:A total 396 patients were evaluated in the study. There were significant differences in the consistency of evaluation among hospitalized, post-operative and discharged patients after intervention compared with before intervention ( P< 0.0167).The consistency of assessment of team members and responsible nurses increased from 68 to 120 cases, and the consistency of assessment reached 90.91%. The Kappa consistency of assessment of responsible nurses and team members increased from 0.354 before intervention to 0.879 after intervention. The effect was remarkable. Conclusions:PDCA cycle method is based on the problems found in clinical practice. It clarifies the professional terms and rules used in the scale, improves the accuracy and consistency of the Autar DVT scale used by nurses, correctly reflects the risk of thrombosis, ensures the safety of patients, and is worthy of promotion and reference.