BackgroundAccurate preoperative identification of high nodal burden (pathologic N2 or N3; hereafter N2+) is important in esophageal squamous cell carcinoma, but contrast-enhanced computed tomography criteria based mainly on nodal size and morphology have limited sensitivity.MethodsIn this retrospective multicohort study, 1,060 consecutive patients with esophageal squamous cell carcinoma who underwent preoperative contrast-enhanced computed tomography and curative-intent esophagectomy with lymphadenectomy were enrolled from two centers. Center A contributed a development cohort (n = 612; train/validation/internal test, 428/92/92) and a temporally held-out cohort (n = 238), and Center B contributed an external test cohort (n = 210). A three-dimensional residual convolutional neural network encoder was pretrained on 3,200 unlabeled chest computed tomography examinations using masked-volume reconstruction and then fine-tuned on tumor-centered volumes comprising the primary tumor plus a 5-mm margin.ResultsThe self-supervised model achieved area under the receiver operating characteristic curve values of 0.881 (95% confidence interval, 0.793–0.955) in the internal test cohort, 0.860 (95% confidence interval, 0.810–0.903) in the temporal cohort, and 0.860 (95% confidence interval, 0.810–0.906) in the external cohort. In the external cohort, sensitivity and specificity at the main operating point were 0.581 and 0.845 for the self-supervised model versus 0.339 and 0.784 for the guideline-inspired comparator. Calibration also improved with self-supervised pretraining (Brier score, 0.148; expected calibration error, 0.053).ConclusionA contrast-enhanced computed tomography-only self-supervised three-dimensional model predicted high pathologic nodal burden in esophageal squamous cell carcinoma with robust temporal and external validation and showed numerically higher performance than a transparent computed tomography-only comparator. Calibrated risk estimates may help prioritize additional nodal workup when staging resources are limited or routine computed tomography findings are equivocal.

Our previous studies have demonstrated that exosomes play a crucial role in promoting vaginal tissue reconstruction in rats. The present study aims to elucidate the molecular mechanisms through which human umbilical cord mesenchymal stem cell-derived exosomes (hUMSC-Exos), which carry microRNA-181a-5p (miR-181a-5p), promote vascularization and tissue regeneration, with particular focus on the involvement of the PTEN/PI3K/AKT signaling pathway. Human umbilical vein endothelial cells (HUVECs) served as a model for studying angiogenesis and cell proliferation, and the expression levels of miR-181a-5p, PTEN, phospho-PI3K (p-PI3K), and phospho-AKT (p-AKT) were analyzed. HUVECs were transfected with PTEN overexpression vector or a negative control vector, then treated with exosomes derived from mesenchymal stem cells (MSCs) transfected with either a miR-181a-5p mimic or an inhibitor. Cell proliferation and migration were assessed using the Cell Counting Kit-8 and scratch assay, respectively, while cell invasion was evaluated via Transwell assay. The StarBase tool was employed to predict binding sites between miR-181a-5p and its target gene, the phosphatase and tensin homolog (PTEN). This interaction was subsequently validated using a dual-luciferase reporter assay. In HUVECs, elevated miR-181a-5p levels were positively correlate with reduced PTEN expression. In vitro experiments demonstrate that hUMSC-Exos enhance HUVEC migration, proliferation, and tube formation. Furthermore, overexpression of PTEN partially counteracted these miR-181a-5p-mediated effects. Our findings indicate that hUMSC-Exos contain miR-181a-5p, which may enhance tube formation and proliferation in HUVECs by regulating PTEN expression, thereby influencing the PI3K/AKT pathway.

Intrauterine adhesions (IUAs) represent a considerable impediment to female reproductive health. Despite ongoing debate regarding the optimally efficacious route of administration and dosage of stem cells for IUA treatment, human umbilical cord-derived mesenchymal stem cells (UCMSCs) have emerged as a promising avenue for regenerative therapy. The present study aimed to investigate the potential effects of UCMSCs on IUAs and to further explore the most effective treatment route and dosages. In the present study, the therapeutic potential of UCMSCs in a constructed rat model of IUAs was evaluated. The efficacy of UCMSC administration through three different routes, namely intraperitoneal injection, in-site injection and caudal vein injection, was compared at three different doses of cells (0.5x106, 1x106 and 5x106). The assessment parameters included endometrial thickness, glandular density and extent of fibrotic tissue, which were measured using HE staining and Masson staining and numbers of offspring. The IUA model group compared with the control group endometrial thickness decreased, glandular density decreased and the extent of fibrotic tissue increased, suggesting the IUA rat model had been successfully established. At 4 weeks post-treatment, an intraperitoneal injection of 1x106 UCMSCs (the middle dose) was found to have led to a significant increase in endometrial thickness and glandular count, approaching the levels that were observed in the normal group. This dosage also notably reduced the level of fibrosis compared with that in both the higher and the lower doses, although this remained slightly higher compared with that observed in the normal group. Furthermore, the reproductive capability of the rats in the higher and middle dosage IUA rat model exhibited partial recovery post-treatment. In conclusion, the results of the present study suggest that the intraperitoneal administration of 1x106 UCMSCs can provide a viable strategy for promoting endometrial regeneration and reducing fibrosis in IUA. In addition, this highlights the potential of UCMSC therapy as a means of clinical intervention for severe IUA, ultimately improving fertility outcomes, especially with regard to the specific dosage and intraperitoneal injection method.

Alopecia areata (AA) is an autoimmune disease characterized by inflammatory and non-scarring hair loss, mediated by CD8+ T cells and primarily affecting hair follicles. Janus kinase (JAK) inhibitors selectively inhibit JAK, block the signal transducer and activator of transcription pathway, and often interfere with T-cell-mediated inflammatory cytokine pathways. They are a class of targeted anti-inflammatory drugs that can promote the activation of hair follicle stem cells. Studies have shown that JAK inhibitors exhibited good efficacy and safety in the treatment of AA, with fewer serious side effects. This article reviews the mechanism of action of JAK inhibitors in the treatment of AA and the effects and side effects of representative drugs.

OBJECTIVE:To investigate the therapeutic effect and underlying mechanism of Guizhi Fuling capsule (GZFL) on bortezomib-induced peripheral neuropathy (BiPN). MATERIALS AND METHODS:Interleukin-6 (IL-6) levels in the plasma of Multiple myeloma (MM) patients were measured by ELISA, and correlation analysis between IL-6 and clinical features of BiPNs was performed. Then, we assess the clinical therapeutic effects of GZFL on MM patients by detecting IL-6 level, PN grade, FACT score, VAS score, MVC and SCV before and after the treatment. A combination of LC/MS and network pharmacology analysis was used to investigate the components and targets of GZFL. Then, bioinformatics was carried out. After PC12 cells were treated with GZFL, a BiPN cell model was constructed to evaluate cell autophagy function by cell viability, IL-6 levels, ROS levels, immunofluorescence staining of LC3 puncta, electron transmission electron microscopy (TEM), and Western blotting (WB). C57BL/6 mice were administered bortezomib by intraperitoneal injection to establish a model of BiPN. Nerve injury in BiPN mice was observed by measuring ethology, motor nerve conduction velocity, and IL-6. ROS, HE staining. TEM, western blotting and IHC were used to detect the expression of autophagy-related indexes. RESULTS:In BiPN patients, IL-6 levels were positively correlated with the PN and FACT, VAS scores. Collectively, GZFL can alleviate BiPN by reducing the level of IL-6, which is mainly manifested in the decline of PN grade, FACT, VAS score and the improvement of MVC and SCV. Thirty-four components and 107 targets of GZFL for BiPN were obtained. IL-6, mTOR, and AKT1 showed high degree values, and the significantly enriched signaling pathways were closely related to inflammatory factors and autophagy pathways, such as TNF and the mTOR signaling pathway. GZFL significantly decreased IL-6 levels in cell and animal models of BiPN. For the autophagy test, GZFL increased PC12 cell ability and the numbers of LC3 puncta and autophagic vesicles after bortezomib treatment. In vivo experiments showed that GZFL effectively improved the behavior of mice with BiPN and alleviated sciatic nerve injury. WB and IHC showed that GZFL enhanced autophagy, as indicated by the alteration of autophagy-related protein levels in PC12 cells and sciatic nerve tissue. CONCLUSION:The present study confirmed that GZFL significantly ameliorates peripheral neuropathy by regulating autophagy levels via alleviating high levels of IL-6 . TRIAL REGISTRATION:The link to the registration: Chinese Clinical Trial Registry (https://www.chictr.org.cn/bin/project/edit?pid=214832). The name of the trial register is "The role of mitochondrial autophagy in multiple myeloma peripheral neuropathy and the application of traditional Chinese medicine for warming Yang and removing blood stasis". The clinical trial registration number is ChiCTR2400088065.

A stereoselective formal (3+2) cycloaddition protocol between 3-methylene isoindolinones and C3-substituted 2-indolylmethanols has been developed. The reactions afforded a wide range of chiral spiro-bis-N-heterocyclic lactams bearing nonadjacent stereocenters, including a quaternary carbon spiro chiral center in good yields (up to 98%) and excellent enantio- and diastereoselectivities (up to >99% ee, >20:1 dr) by chiral phosphoric acid organocatalysis (0.5 mol %). Gram-scale reactions and diversified synthetic transformations of the desired spiro-lactams without loss of stereochemical purity further substantiate their potential utility.

OBJECTIVES:To investigate the antitumor effects of aucubin (AC) in non-small cell lung cancer (NSCLC) and uncover its plausible mechanism against lung cancer stem-like cells (LCSCs). METHODS:In vitro experiments included MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, a reagent commonly used for cell viability assay) and colony formation assays to assess anti-proliferative effects on A549 and NCI-H1975 lung cancer cell lines, wound healing and Transwell invasion assays to evaluate inhibition of cell migration and invasion, tumorsphere-formation experiments to detect changes in NSCLC cell stemness, as well as Western blot and quantitative reverse transcription polymerase chain reaction (qRT-PCR) analyses to measure the expression of LCSC markers (CD44, CD133, Oct4, and Nanog). In vivo experiments were conducted to observe the impact of AC on NSCLC metastasis and mouse survival rates. Further mechanistic studies involved transcriptomic gene set enrichment analysis, Western blot, qRT-PCR, molecular docking, and Surface Plasmon Resonance (SPR) methods to investigate how AC directly targets β-catenin and promotes its ubiquitin-mediated degradation. KEY FINDINGS:AC exerted significant anti-proliferative effects on A549 and NCI-H1975 cells, inhibited cancer cell migration and invasion, reduced the stemness of NSCLC cells, and markedly downregulated the expression of LCSC markers in vitro. In vivo, AC treatment significantly reduced NSCLC metastasis and improved mouse survival rates.Mechanistically, AC blocked the WNT (Wingless-related integration site, a family of secreted lipid-modified signaling glycoproteins that play crucial roles in embryonic development, tissue homeostasis, and regeneration) signaling pathway by downregulating β-catenin and c-Myc expression. It directly targeted β-catenin, promoting its degradation via the ubiquitin-proteasome pathway. CONCLUSIONS:This study uncovers a novel anti-LCSC mechanism of AC, offers alternative strategies for NSCLC treatment, and provides innovative lead compounds for the development of drugs targeting lung cancer stem cells.

Background: To explore the antioxidant capacity of alpha-asarone (ARE) in mice models of sodium valproate (SV)-induced liver injury, and to elucidate the underlying mechanism of ARE in liver injury treatment. Methods: Network pharmacology and molecular docking were used to predict ARE's potential pharmacological mechanisms in treating drug-induced liver injury. AML12 cell model was used to evaluate the antioxidant stress activation of ARE. In the evaluation of antioxidant activity, ARE was tested for its ability to scavenge hydroxyl radical (OH) radicals, 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+), and 1,1-diphenyl-2-picrylhydrazyl (DPPH), with vitamin C serving as the positive control. And thirty-six male mice were divided into six groups. Group I, the control, received saline. The other five groups received oral doses of 500 mg/kg/b.w. of SV daily for 14 days. Group II received only SV. Group III received the hepatic protectant bifendate at 100 mg/kg/b.w., while groups IV, V, and VI were treated with 20, 40, and 80 mg/kg/b.w. of ARE, respectively, also for 14 days. After sacrifice, liver and blood samples were collected for biochemical or metabolomic analysis. Results: ARE significantly reduced the levels of reactive oxygen species and activated the Keap1/Nrf2 signaling pathway in liver injury AML12 cells. ARE demonstrated dose-dependent scavenging activity in DPPH, ABTS+, and OH assays. It lowered serum alanine aminotransferase and aspartate aminotransferase levels, increased glutathione peroxidase and superoxide dismutase activity in liver tissue, and reduced malondialdehyde content. Metabolomic analysis showed that ARE restored twenty liver metabolites to control levels and enriched three pathways (arginine biosynthesis, sucrose and starch metabolism, and biotin metabolism) with shared differential metabolites. Conclusion: These results shed light on ARE's mechanism in reducing oxidative stress, suggesting a potential strategy for preventing SV-induced liver disease by modulating metabolic products and enhancing antioxidant capacity. ARE could be a promising drug candidate for its antioxidant and liver-protective effects.

A stable isotope dilution-liquid chromatography tandem mass spectrometry method based on a low-temperature derivatization strategy with 3-nitrophenylhydrazine (3-NPH) was developed for the determination of six volatile fatty acids (VFAs) in serum, urine, and feces. Ice acetonitrile was used to precipitate proteins and extract the target analytes. The extract was derivatized with 3-NPH methanol solution at 4 degrees C. BEH C8 (1.7 mu m, 2.1 x 100 mm) column was used for chromatographic separation, and acetonitrile-water (both containing 0.01 % formic acid) were used as the mobile phase with a gradient elution of 10 min. Electrospray ionization source (ESI) in negative ion multiple reaction monitoring (MRM) mode were used for analyte detection. The regression coefficients R2 of the calibration curves for the six VFAs were in the range of 0.9963-0.9994, and the LOQs were in the range of 0.02-0.5 mu g mL-1, with the recoveries in the range of 85.3-104.3 %, and the intra- and inter-day precision in the range of 1.8-9.1 %. The method is simple, accurate and reliable, and has been applied in the sensitive determination of VFAs in complex biological samples.

[This corrects the article on p. 1391 in vol. 11, PMID: 33948364.].

Human mesenchymal stem cells (hMSCs) are mesoderm-derived adult stem cells with self-proliferation capacity, pluripotent differentiation potency, and excellent histocompatibility. These advantages make hMSCs a promising tool in clinical application. However, the majority of clinical trials using hMSC therapy for diverse human diseases do not achieve expectations, despite the prospective pre-clinical outcomes in animal models. This is partly attributable to the intrinsic heterogeneity of hMSCs. In this review, the cause of heterogeneity in hMSCs is systematically discussed at multiple levels, including isolation methods, cultural conditions, donor-to-donor variation, tissue sources, intra-tissue subpopulations, etc. Additionally, the effect of hMSCs heterogeneity on the contrary role in tumor progression and immunomodulation is also discussed. The attempts to understand the cellular heterogeneity of hMSCs and its consequences are important in supporting and improving therapeutic strategies for hMSCs.

Unintentional, early pregnancy alcohol consumption affects embryonic development. During the peri-implantation stage, coinciding with the transition from naive to primed pluripotency, the long isoform of KDM2B (KDM2BLF) underlies the de novo establishment of polycomb repressive complex (PRC) functions at promoters after fertilization. However, it remains unclear whether and how ethanol exposure affects this spatiotemporal chromatin setting. Here, we show that exposing peri-implantation mouse embryos to ethanol leads to impaired post-implantation development, mirrored by the delayed exit of naive pluripotency in acetaldehyde-treated embryonic stem cells. Remarkably, these abnormalities are linked to inadequate KDM2BLF expression and compromised deposition of PRC marks, which arise from cAMP response element-binding protein (CREB) inactivation. Accordingly, pharmacological activation of CREB effectively restores pluripotency transition partly dependent on KDM2BLF in vitro and ameliorates post-implantation embryonic defects in vivo. Therefore, our study highlights the pivotal role of the CREB/KDM2B axis in chromatin configuration and developmental programming, proposing potential preventive strategies against ethanol exposure-induced detrimental effects.

DNA damage is a key factor affecting gametogenesis and embryo development. The integrity and stability of DNA are fundamental to a woman’s successful conception, embryonic development, pregnancy and the production of healthy offspring. Aging, reactive oxygen species, radiation therapy, and chemotherapy often induce oocyte DNA damage, diminished ovarian reserve, and infertility in women. With the increase of infertility population, there is an increasing need to study the relationship between infertility related diseases and DNA damage and repair. Researchers have tried various methods to reduce DNA damage in oocytes and enhance their DNA repair capabilities in an attempt to protect oocytes. In this review, we summarize recent advances in the DNA damage response mechanisms in infertility diseases such as PCOS, endometriosis, diminished ovarian reserve and hydrosalpinx, which has important implications for fertility preservation.

Due to continuous application as a flavoring agent in the pesticide, pharmaceutical, and food industries, methyl eugenol (ME) persists in the environment and causes deleterious impacts including cytotoxicity, genotoxicity, and liver damage. This study utilized a comprehensive approach, integrating toxicokinetics, metabolomics, and gut microbiota analysis, to explore the mechanisms behind ME-induced hepatotoxicity in mice. The study observed significant rises in ALT and AST levels, along with significant weight loss, indicating severe liver damage. Toxicokinetic data showed delayed Tmax and plasma accumulation after 28 days of repeated ME exposure at doses of 20 mg/kg, 40 mg/kg, and 60 mg/kg. The metabolomic analysis pinpointed four critical pathways—TCA cycle; alanine, aspartate, and glutamate metabolism; arginine biosynthesis; and tyrosine metabolism—linked to 20 potential biomarkers. Gut microbiota analysis revealed that extended ME exposure led to microbial imbalance, particularly altering the populations of Akkermansia, Prevotella, and Ruminococcus, which are key to amino acid metabolism and the TCA cycle, thus contributing to hepatotoxicity. However, the causal relationship between changes in gut microbiota and liver metabolite levels still requires further in-depth research. This study underscores the significant role of liver metabolites and gut microbiota in ME-induced liver damage.

Background: Studies have shown that the chronic use of cannabis is associated with a decrease in blood pressure. Our previous studies prove that activating the cannabinoid type 2 (CB2) receptor in the brain can effectively reduce blood pressure in spontaneously hypertensive rats; however, the exact mechanism has not been clarified. The objective of this study is to demonstrate that activation of microglial CB2 receptors can effectively reduce the levels of TNF-α, IL-1β, and IL-6 in the paraventricular nucleus (PVN) through inhibiting aerobic glycolysis, thereby relieving hypertension. Methods: AngiotensinII (AngII) was administered to BV2 cells and C57 mice to induce hypertension and the release of proinflammatory cytokines. The mRNA and protein expression of the CB2 receptor, TNF-α, IL-1β, IL-6, and the PFK and LDHa enzymes were detected using RT-qPCR and Western blotting. The Seahorse XF Energy Metabolism Analyzer was used to measure the oxidative phosphorylation and aerobic glycolysis metabolic pathways in BV2 cells. The long-term effects of injecting JWH133, a selective CB2 receptor agonist, intraperitoneally on blood pressure were ascertained. ELISA was used to measure norepinephrine and lactic acid levels while immunofluorescence labeling was used to locate the CB2 receptor and c-Fos. By injecting pAAV-F4/80-GFP-mir30shRNA (AAV2-r-CB2shRNA) into the lateral cerebral ventricle, the CB2 receptor in microglia was specifically knocked down. Results: Activation of CB2 receptors by the agonist JWH133 suppressed TNF-α, IL-1β, and IL-6 by inhibiting PFK and LDHa enzymes involved in glycolysis, as well as lactic acid accumulation, along with a reduction in glycoPER levels (marks of aerobic glycolysis) in AngII-treated BV2 cells. In AngII-treated mice, the administration of JWH133 specifically activated CB2 receptors on microglia, resulting in decreased expression levels of PFK, LDHa, TNF-α, IL-1β, and IL-6, subsequently leading to a decrease in c-Fos protein expression within PVN neurons as well as reduced norepinephrine levels in plasma, ultimately contributing to blood pressure reduction. Conclusion: The results suggest that activation of the microglia CB2 receptor decreases the neuroinflammation to relieve hypertension; the underlying mechanism is related to inhibiting aerobic glycolysis of microglia.

Background To compare the expression levels of long non-coding RNA (lncRNA) and messenger RNA (mRNA) in pre-receptive endometrium between patients with Polycystic Ovary Syndrome (PCOS)and normal ovulation undergoing in vitro fertilization-embryo transfer (IVF-ET). Methods Endometrial tissues were collected with endometrial vacuum curette in pre-receptive phase (3 days after oocytes retrieval) from PCOS and control groups. LncRNAs and mRNAs of endometrium were identified via RNA sequencing and alignments. A subset of 9 differentially expressed lncRNAs and 11 mRNAs were validated by quantitative reverse transcription polymerase chain reaction(qRT-PCR)in 22 PCOS patients and 18 ovulation patients. The function of mRNAs with differential expression patterns were explored using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Results We found out 687 up-regulated and 680 down-regulated mRNAs, as well as 345 up-regulated and 63 down-regulated lncRNAs in the PCOS patients in contrast to normal ovulation patients. qRT-PCR was used to detect the expression of 11 mRNAs, and validated that the expression of these 6 mRNAs CXCR4, RABL6, OPN3, SYBU, IDH1, NOP10 were significantly elevated among PCOS patients, and the expression of ZEB1 was significantly decreased. qRT-PCR was performed to detect the expression of 9 lncRNAs, and validated that the expression of these 7 lncRNAs IDH1-AS1, PCAT14, FTX, DANCR, PRKCQ-AS1, SNHG8, TPT1-AS1 were significantly enhanced among PCOS patients. Bioinformatics analysis showed that differentially expressed genes (DEGs) involved KEGG pathway were tyrosine metabolism, PI3K-Akt pathway, metabolic pathway, Jak-STAT pathway, pyruvate metabolism, protein processing in endoplasmic reticulum, oxidative phosphorylation and proteasome. The up-regulation of GO classification was involved in ATP metabolic process, oxidative phosphorylation, RNA catabolic process, and down-regulation of GO classification was response to corticosteroid, steroid hormone, and T cell activation. Conclusion Our results determined the characteristics and expression profile of endometrial lncRNAs and mRNAs in PCOS patients in pre-receptive phase, which is the day 3 after oocytes retrival. The possible pathways and related genes of endometrial receptivity disorders were found, and those lncRNAs may be developed as a predictive biomarker of endometrium in pre-receptive phase.

PurposeThe objective of this study is to assess the carrier frequency and pathogenic variation of monogenetic diseases in a population of 114 subjects in Han Chinese from Hebei province who are undergoing assisted reproductive technology through the utilization of Expanded Carrier Screening (ECS).MethodsThe study utilized a panel consisting of 155 severe monogenic recessive genetic diseases for ECS. Next-generation sequencing technology was employed to identify specific variants associated with ECS in a cohort of 114 subjects from 97 couples, comprising 97 females and 17 male spouses.ResultsA total of 114 individuals received ECS. The carrier rate of pathogenic genes in the enrolled population was 44.74% (51/114). Among the 97 females, the carrier rate of pathogenic genes was higher in those without assisted reproduction indicators than in those with assisted reproduction indicators (59.09% vs. 41.33%). However, the carrier rate of pathogenic genes in males without assisted reproductive technology was slightly lower than that with assisted reproductive technology (40% vs. 41.67%). Among both female and male participants, the carrier rate of pathogenic genes between individuals without indicators of assisted reproduction and those with such indicators was 55.55% vs. 41.38%. In 51 carriers, 72.55% (37/51) carried one genetic variant, 25.49% (13/51) carried two genetic variants, and 1.96% (1/51) carried three genetic variants. A total of 38 pathogenic genes were detected in this study, and GJB2 and MMACHC were most common. The carrier rates of the two genes were both 5.26% (6/114). A total of 55 variations were detected, and c.235delC was most frequently found. The carrier rate was 3.51% (4/114). The incidence of couples carrying the same pathogenic genes was 1.03% (1/97).ConclusionsThe findings elucidate the carrier rate of pathogenic genes among 155 severe monogenic recessive genetic diseases and underscore the significance of ECS as a preventive measure against congenital anomalies. When both partners carry the same genetic mutation for a monogenic disease, preventive strategies can be taken in offspring through preimplantation genetic testing (PGT), prenatal genetic testing, or the utilization of donor gametes. ECS is instrumental in assessing reproductive risk, guiding fertility-related decisions, and reducing the prevalence of monogenic recessive genetic disorders in subsequent generations.

Tetrandrine (TET), a natural bisbenzyl isoquinoline alkaloid extracted from Stephania tetrandra S. Moore, has diverse pharmacological effects. However, its effects on melanoma remain unclear. Cellular proliferation assays, multi-omics analyses, and xenograft models were used to determine the effect of TET on melanoma. The direct target of TET was identified using biotin-TET pull-down liquid chromatograph-mass spectrometry (LC-MS), cellular thermal shift assays, and isothermal titration calorimetry (ITC) analysis. Our findings revealed that TET treatment induced robust cellular autophagy depending on activating transcription factor 6 (ATF6)-mediated endoplasmic reticulum (ER) stress. Simultaneously, it hindered autophagic flux by inducing cytoskeletal protein depolymerization in melanoma cells. TET treatment resulted in excessive accumulation of reactive oxygen species (ROS) and simultaneously triggered mitophagy. Sirtuin 5 (SIRT5) was ultimately found to be a direct target of TET. Mechanistically, TET led to the degradation of SIRT5 via the ubiquitin (Ub)-26S proteasome system. SIRT5 knockdown induced ROS accumulation, whereas SIRT5 overexpression attenuated the TET-induced ROS accumulation and autophagy. Importantly, TET exhibited anti-cancer effects in xenograft models depending on SIRT5 expression. This study highlights the potential of TET as an antimelanoma agent that targets SIRT5. These findings provide a promising avenue for the use of TET in melanoma treatment and underscore its potential as a therapeutic candidate.

Abstarct Background 3D-printing is widely used in regenerative medicine and is expected to achieve vaginal morphological restoration and true functional reconstruction.Mesenchymal stem cells-derived exosomes (MSCs-Exos) were applyed in the regeneration of various tissues.The current study aimed to explore the effctive of MSCs-Exos in vaginal reconstruction. Results In this work, hydrogel was designed using decellularized extracellular matrix (dECM) and gelatin methacrylate (GelMA) and silk fibroin (SF).The biological scaffolds was constructed using desktop-stereolithography.The physicochemical properties of the hydrogels were evaluated.It was observed that the sustained release property of exosomes in the hydrogel both in vitro and in vitro.The results revealed that 3D scaffold encapsulating exosomes expressed significant effects on the vascularization and musule regeneration of the regenerative vagina tissue.Also, MSCs-Exos strongly promoted vascularization in the vaginal reconstruction of rats,which may through the PI3K/AKT signaling pathway. Conclusions Our results indicated that the 3D-printed, lumenal scaffold encapsulating exosomes might be used as a cell-free alternative treatment strategy for vaginal reconstruction.