Surgical Treatment of Multiple Large Tuberous and Tendinous Xanthoma Secondary to Familial Hypercholesterolaemia: A Case Report.

Xanthomas are well-circumscribed skin lesions that are commonly seen in patients with familial hypercholesterolemia (FH). The aim of this report is to present a rare case of multiple large tuberous and tendinous xanthomas. A 17-year-old female patient in this report presented with multiple asymptomatic and papulo-nodular masses in both sides of palms, elbows, buttocks, knees, and Achilles tendons. Surgical removal of the masses was carried out in combination with lipid-lowering therapy. A following up of 3 months showed all wounds were healing well, and no recurrence of masses was observed. Therefore, for patients with xanthomas related with familial hypercholesterolaemia, lipid-lowering therapy has reportedly reduced the size of masses, but surgical treatment may be essential for large xanthomas caused pain or limitation of daily activities.

Chitosan Hydrogel Dressing Loaded with Adipose Mesenchymal Stem Cell-Derived Exosomes Promotes Skin Full-Thickness Wound Repair.

Cutaneous trauma repair is still a challenge in the clinic due to the scar formation and slow healing rate, especially for diabetic patients. Various drug-loading wound dressings have been explored to solve this problem. Mesenchymal stem cell (MSC)-derived exosomes have been considered as a potential cell-free drug because of their anti-inflammation function and tissue repair property that are comparable to that of MSCs. Herein, a composite wound dressing (Exo/Gel) consisting of the chitosan hydrogel and adipose MSC-derived exosome (ADMSC-Exo) was designed and fabricated by a physical mixing method to promote the skin full-thickness wound repair. The exosomes were slowly released from the Exo/Gel dressing with the degradation of the chitosan hydrogel. The Exo/Gel displayed enhanced cell migration and angiogenic properties in vitro. And the results in the rat skin wound model showed that the Exo/Gel could promote the regular collagen deposition, angiogenesis, and hair follicle mosaicism regeneration. These results proved that the hydrogel dressing with ADMSCs-derived exosomes can accelerate skin wound healing, which is a strategy for developing wound dressings.

The Effects of Intravenous Dexamethasone on Rebound Pain After Nerve Block in Patients with Ankle Fracture: A Randomized Controlled Trial.

Purpose: A single-injection nerve block provides excellent analgesia in a short time, but rebound pain after the nerve block disappears has attracted researchers' attention. The aim of this study is to evaluate the effect of intravenous dexamethasone on rebound pain after adductor canal block (ACB) and popliteal sciatic nerve block in patients with ankle fracture. Methods: We recruited 130 patients with ankle fractures scheduled for open reduction and internal fixation (ORIF), each of whom received ACB and popliteal sciatic nerve block. Patients were divided into two groups: C (ropivacaine only) and IV (ropivacaine with intravenous dexamethasone). The primary outcome was the incidence of rebound pain. Secondary outcomes included the following: pain scores at 6 h (T1), 12 h (T2), 18 h (T3), 24 h (T4), and 48 h (T5) after operation; duration of the nerve block; number of presses of the analgesia pump and rescue analgesic consumption in the three-day postoperative period; quality of recovery scale (QoR-15 score); postoperative sleep quality; satisfaction of patients; and levels of serum inflammatory markers (IL-1ß, IL-6, and TNF-α) six hours after surgery. Results: Compared with group C, the incidence of rebound pain in group IV was significantly reduced, and the duration of nerve block was extended by approximately nine hours (P<0.05). Moreover, patients in group IV had significantly lower pain scores at T2-T4, lower levels of serum inflammatory markers (IL-1ß, IL-6, and TNF-α), higher QoR-15 score two days after the operation, and satisfactory sleep quality the night after surgery (P<0.05). Conclusion: Intravenous dexamethasone can reduce the rebound pain after adductor block and sciatic popliteal nerve block in patients with ankle fracture surgery, prolong the duration of nerve block, and improve the quality of early postoperative recovery.

Efficacy of Femoral Neck System Fixation vs KHS Femoral Neck Dynamic Compression Locking Plate System in Femoral Neck Fracture.

Background: Femoral neck fracture is acknowledged as one of the common injuries in clinical orthopedics. Our study was aimed at investigating the efficacy of femoral neck fixation vs the KHS dynamic compression locking plate system in the treatment of femoral neck fracture. Methods: This was a prospective study. A toteal of 90 patients with femoral neck fracture who were admitted to The Third Hospital of Hebei Medical University in Shijiazhuang, China from August 2017 to March 2020 were enrolled in our study. The patients were randomly divided into the control group (45 patients allocated to intervention with the novel femoral neck dynamic compression locking plate system) and the study group (45 patients who underwent femoral neck system fixation). Intraoperative blood loss, surgery duration, fracture healing time and related complications in the 2 groups were monitored and evaluated. The recovery of hip joint function at different times in the 2 groups were closely monitored. Results: The 2 groups completed the surgery process, and the incision healed. All patients were followed up for 6 to 8 months, with an average follow-up time of 7.01 ± 0.21 months. Surgery duration, length of hospital stay and fracture healing time in the study group were significantly lower than in the control group (P < .05), while no significant difference was found in intraoperative blood loss between the 2 groups (P > .05). At 1 and 3 months after surgery, hip joint function in the study group was significantly higher than in the control group (P < .05), but 6 months after surgery, there was no significant difference between the 2 groups (P > .05). There were no complications in the study group, whereas 1 patient had a complication in the control group. The total incidence of complications in the study group was lower than in the control group, but the difference was not significant (P > .05). Conclusion: Femoral neck system fixation demonstrated superior efficacy to the KHS femoral neck dynamic compression locking plate system in femoral neck fracture, and is considered as a valid method for wide application.

Selenium-enriched Bacillus subtilis Improves Growth Performance, Antioxidant Capacity, Immune Status, and Gut Health of Broiler Chickens.

This study aimed to investigate the effects of selenium (Se)-enriched Bacillus subtilis (Se-BS) on growth performance, antioxidant capacity, immune status, and gut health in broilers. A total of 240 one-day-old Arbor Acres broilers were randomly allotted to four groups and fed with basal diet (control group), 0.30 mg/kg Se (SS group), 3 × 109 CFU/g B. subtilis (BS group), and 0.30 mg/kg Se + 3 × 109 CFU/g B. subtilis (Se-BS group) for 42 days. The results showed that Se-BS supplementation increased body weight (BW), average daily gain, the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), and peroxidase (POD), total antioxidant capacity (T-AOC), and the contents of interleukin (IL)-2, IL-4, and immunoglobulin (Ig) G in plasma, the index and wall thickness of the duodenum, the villus height and crypt depth of the jejunum, and GPx-1 and thioredoxin reductase 1 mRNA levels in liver and intestine and decreased feed conversion ratio (FCR) and plasma malondialdehyde (MDA) content compared with the control group on day 42 (P < 0.05). Compared with groups SS and BS, Se-BS supplementation increased BW, the activities of GPx, CAT, and POD, and the contents of IL-2, IL-4, and IgG in plasma, the index and wall thickness of the duodenum, the crypt depth and secretory IgA content of the jejunum, and GPx-1 mRNA levels in liver and intestine and decreased FCR and plasma MDA content on day 42 (P < 0.05). In conclusion, Se-BS supplementation effectively improved the growth performance antioxidant capacity, immune status, and gut health of broilers.

MiR-7-5p/KLF4 signaling inhibits stemness and radioresistance in colorectal cancer.

Resistance to radiotherapy remains a major unmet clinical obstacle in the treatment of locally advanced rectal cancer. Cancer stem cells (CSCs) are considered to mediate tumor development and radioresistance. However, the role of CSCs in regulating resistance to radiotherapy in colorectal cancer (CRC) remains largely unknown. We established two radioresistant CRC cell lines, HCT116-R and RKO-R, using fractionated irradiation. Analysis using miRNA sequencing and quantitative real-time PCR confirmed lower levels of miR-7-5p in both of the radioresistant cells compared to their parental cells. Subsequently, we validated that miR-7-5p expression was decreased in cancerous tissues from radiotherapy-resistant rectal cancer patients. The Cancer Genome Atlas (TCGA) database analyses revealed that low miR-7-5p expression was significantly correlated with poor prognosis in CRC patients. Overexpression of miR-7-5p led to a rescue of radioresistance and an increase in radiation-induced apoptosis, and attenuated the stem cell-like properties in HCT116-R and RKO-R cells. Conversely, knocking down miR-7-5p in parental HCT116 and RKO cells suppressed the sensitivity to radiation treatment and enhance cancer cell stemness. Stemness-associated transcription factor KLF4 was demonstrated as a target of miR-7-5p. Rescue experiments revealed that miR-7-5p/KLF4 axis could induce radiosensitivity by regulating CSCs in colorectal cancer cells. Furthermore, we used CRC tumor tissues which exhibited resistance to neoadjuvant radiotherapy to establish a patient-derived xenograft (PDX) mouse model. Tail vein injection of magnetic nanoparticles carrying miR-7-5p mimics into the PDX mice significantly inhibited tumor growth with or without irradiation treatment in vivo. Our current studies not only demonstrate an anti-cancer function of miR-7-5p in regulating CSC properties and radiosensitivity in colorectal cancer, but also provide a novel potential strategy for delaying or reverse radiation resistance in preoperative radiotherapy of CRC patients.

A fluorescent sensing platform based on collagen peptides-protected Au/Ag nanoclusters and WS2 for determining collagen triple helix integrity.

The unique triple helix structure of collagen plays an important role in its biological properties, and the triple helix integrity is closely correlated with its molecular behavior and biological functions. Nevertheless, there is still a lack of convenient, accurate and practical methods for quantitatively determining collagen triple helix integrity. Herein, we first prepared bovine skin collagen peptide (BSCP)-protected Au/Ag nanoclusters (Au/AgNCs@BSCP) with excellent optical properties, high stability and good biocompatibility, which could adsorb on WS2 surface leading to fluorescence quenching. Upon the addition of collagen, the interaction of collagen and Au/AgNCs@BSCP led to the detachment of Au/AgNCs@BSCP from the WS2 surface, causing an increase in the fluorescence signal. Using the difference in the fluorescence recovery of the different samples, we achieved the quantitative determination of collagen triple helix integrity. This developed strategy exhibited excellent accuracy, selectivity, and practicality, thus showing promising potentials in biomedical applications.

Mesenchymal Stem Cell Derived Extracellular Vesicles: Promising Nanomedicine for Cutaneous Wound Treatment.

A skin wound represents a rupture caused by external damage or the existence of underlying pathological conditions. Sometimes, skin wound healing processes may place a heavy burden on patients, families, and society. Wound healing processes mainly consist of several continuous, dynamic, but overlapping stages, namely, the coagulation stage, inflammation stage, proliferation stage, and remodeling stage. Bacterial infection, excessive inflammation, impaired angiogenesis, and scar formation constitute the four significant factors impeding the recovery efficacy of skin wounds. This encourages scientists to develop multifunctional nanomedicines to meet challenging needs. As we know, mesenchymal stem cells (MSCs) have been widely explored for wound repair owing to their unique capability for self-renewal and multipotency. However, problems including immune concerns and legal restrictions should be properly resolved before MSC-based therapeutics are safely and widely used in clinics. Besides, maintaining the high viability/proliferation capability of MSCs during administration processes and therapy procedures is also one of the biggest technical bottlenecks. Extracellular vesicles (EVs) are cell-derived nanovesicles, that not only possess the basic characteristics and functions of their corresponding maternal cells but also contain several outstanding advantages including abundant sources, excellent biocompatibility, and convenient administration routes. Furthermore, the membrane surface and cavity are easy to flexibly modify to meet versatile application needs. Recently, MSC-derived EVs have emerged as promising therapeutics for skin wound repair. However, current reviews are too broad and rarely focused on the specific roles of EVs in the different stages of wound recovery. Therefore, it is quite necessary to demonstrate the significance of stem cell-derived EVs in promoting wound healing from several specific aspects. Here, this review primarily tries to provide critical comments on current advances in EVs derived from MSCs for wound repair, particularly elaborating on their impressive roles in effectively eliminating infections, inhibiting inflammation, promoting angiogenesis, and reducing scar formation. Last but not least, current limitations and future prospects of EVs derived from MSCs in the areas of wound repair are also objectively analyzed.

An injectable hydrogel scaffold with IL-1ß-activated MSC-derived exosomes for the treatment of endometritis.

Chronic endometritis is a common gynecological disease resulting from various long-term recurrent infections, and is closely related to myositis, miscarriage, and even infertility. There is still no satisfactory treatment method currently in clinical therapy. Mesenchymal stem cell (MSC)-derived exosomes, an important kind of paracrine product, have been used to treat inflammatory diseases due to their promising immunomodulatory function and tissue repair ability similar to MSCs. Considering that the exosome contents and functions are regulated by the MSC status and the MSC status is significantly influenced by its surrounding microenvironment, we propose a hypothesis that exosomes derived from inflammation-simulated MSCs will possess stronger inhibition ability for inflammation. Herein, we used IL-1ß to activate rat bone MSCs for obtaining ß-exo and constructed an injectable polypeptide hydrogel scaffold by loading ß-exo (ß-exo@pep) for an in situ slow release of ß-exo. The results showed that the polypeptide hydrogel can provide a sustained release of exosomes in 14 days. The ß-exo@pep composite hydrogel can more effectively inhibit the production of inflammatory factors such as TNF-α, IL-1ß, and IFN-γ, while it can promote the production of anti-inflammatory factors such as Arg-1, IL-6, and IL-10. The ß-exo@pep composite hydrogel significantly promoted cell migration, invasion, and vessel tube formation in vitro. The experiments in a rat model of endometritis proved that the ß-exo@pep composite scaffold possessed excellent ability towards anti-inflammation and endometrial regeneration. The research studies on the molecular mechanism revealed that the protein expressions of HMGB1 and phosphorylated IKB-α and p65 are down-regulated in the cells treated with ß-exo@pep, indicating the involvement of the NF-κB signaling pathway. This study provides an effective method for the treatment of chronic endometritis, which is promising for clinical use.

Stem cell-derived exosomes for chronic wound repair.

Stem cells possess the capability of self-renewal and multipotency, which endows them with great application potential in wound repair fields. Yet, several problems including immune concerns, ethical debates, and oncogenicity impede the broad and deep advance of stem cell-based products. Recently, owing to their abundant resources, excellent biocompatibility, and ease of being engineered, stem cell-derived exosomes were proved to be promising nanomedicine for curing chronic wounds. What is more, stem cell-derived exosomes are almost the mini record of their maternal cells, which even equipped them with the unique characteristics of stem cells. Chronic wound healing efficacy is dominated by several complicated factors, especially the excessive inflammation conditions and impaired vessels. Therefore, this review tries to concentrate on the current advances of stem cell-derived exosomes for reducing inflammation and promoting angiogenesis in chronic wound healing processes. Last but not least, the existing limitations and future perspectives of stem cell-derived exosomes for chronic wound treatment are also outlined.

Tripartite motif-containing protein 31 confers protection against nonalcoholic steatohepatitis by deactivating mitogen-activated protein kinase kinase kinase 7.

BACKGROUND AIMS: As a global health threat, NASH has been confirmed to be a chronic progressive liver disease that is strongly associated with obesity. However, no approved drugs or efficient therapeutic strategies are valid, mainly because its complicated pathological processes is underestimated. APPROACH RESULTS: We identified the RING-type E3 ubiquitin transferase-tripartite motif-containing protein 31 (TRIM31), a member of the E3 ubiquitin ligases family, as an efficient endogenous inhibitor of transforming growth factor-beta-activated kinase 1 (mitogen-activated protein kinase kinase kinase 7; MAP3K7), and we further confirmed that TRIM31 is an MAP3K7-interacting protein and promotes MAP3K7 degradation by enhancing ubiquitination of K48 linkage in hepatocytes. Hepatocyte-specific Trim31 deletion blocks hepatic metabolism homeostasis, concomitant with glucose metabolic syndrome, lipid accumulation, up-regulated inflammation, and dramatically facilitates NASH progression. Inversely, transgenic overexpression, lentivirus, or adeno-associated virus-mediated Trim31 gene therapy restrain NASH in three dietary mice models. Mechanistically, in response to metabolic insults, TRIM31 interacts with MAP3K7 and conjugates K48-linked ubiquitination chains to promote MAP3K7 degradation, thus blocking MAP3K7 abundance and its downstream signaling cascade activation in hepatocytes. CONCLUSIONS: TRIM31 may serve as a promising therapeutic target for NASH treatment and associated metabolic disorders.

Combined femoral artery block and femoral nerve block reduces thigh tourniquet-induced hypertension.

STUDY OBJECTIVE: Tourniquet hypertension (TH) is thought to be caused by sympathetically mediated C-fibers in the femoral epicardium following prolonged intraoperative inflation of the tourniquet, and we hypothesized that blocking the femoral artery at the same time as a conventional femoral nerve block would reduce the incidence of TH. DESIGN: A prospective, double-blind, randomized, controlled trial. SETTING: Operating room and hospital ward in the Third Hospital of Hebei Medical University. PATIENTS: A total of 72 patients receiving high tibial osteotomy under general anesthesia were recruited from June 2022 to September 2022. INTERVENTIONS: Patients were randomly assigned to receive either a classical femoral nerve block (CFNB) or a modified femoral nerve block (MFNB). Patients in the CFNB group received a 30 mL of 0.5% ropivacaine femoral nerve block and patients in the MFNB group received a 20 mL of 0.5% ropivacaine femoral nerve block combined with a 10 mL of 0.5% ropivacaine femoral artery block. MEASUREMENTS: The primary outcome assessed was the incidence of TH. Data on intraoperative esmolol dosage, analgesic effect, complications and hemodynamics during surgery were also recorded. MAIN RESULTS: Incidence of TH was significantly higher in the CFNB group compared with the MFNB group (71.88% vs 31.25%, P = 0.002). The systolic blood pressure in the CFNB group was significantly higher than that in the MFNB group at 45, 60, 75 and 90 min after tourniquet inflation (P = 0.029, P = 0.020, P = 0.009, P = 0.007). There was also a significant increase in intraoperative esmolol dosage in the CFNB group (65.63 ± 44.15 vs 22.19 ± 33.74, P < 0.001). Postoperative pain scores and patient satisfaction were not statistically significant between the two groups. CONCLUSIONS: The present study demonstrated that modified femoral nerve block reduced intraoperative esmolol dosage and the incidence of TH.

Characterization of immune landscape and development of a novel N7-methylguanine-related gene signature to aid therapy in recurrent aphthous stomatitis.

OBJECTIVES: Recurrent aphthous stomatitis (RAS) is the most common inflammatory disease of the oral mucosa resulting in an impaired life quality and even leading to tumors in susceptible populations. N7-Methylguanine (m7G) plays a vital role in various cellular activities but has not yet been investigated in RAS. We aimed at picturing the immune landscape and constructing an m7G-related gene signature, and investigating candidate drugs and gene-disease association to aid therapy for RAS. METHODS: For our study, m7G-related differentially expressed genes (DEGs) were screened. We outlined the immune microenvironment and studied the correlations between the m7G-related DEGs and immune cells/pathways. We performed functional enrichment analyses and constructed the protein-protein interaction (PPI) and multifactor regulatory network in RAS. The m7G-related hub genes were extracted to formulate the corresponding m7G predictive signature. RESULTS: We obtained 11 m7G-related DEGs and studied a comprehensive immune infiltration landscape, which indicated several immune markers as possible immunotherapeutic targets. The PPI and multifactor regulatory network was constructed and 4 hub genes (DDX58, IFI27, IFIT5, and PML) were identified, followed by validation of the corresponding m7G predictive signature for RAS. GO and KEGG analyses revealed the participation of JAK-STAT and several immune-related pathways. Finally, we suggested candidate drugs and gene-disease associations for potential RAS medical interventions. CONCLUSIONS: The present study pictured a comprehensive immune infiltration landscape and suggested that m7G played a vital role in RAS through immune-related pathways. This study provided new insight for the future investigation of the mechanisms and therapeutic strategies for RAS.

Nomograms and scoring system for forecasting overall and cancer-specific survival of patients with prostate cancer.

BACKGROUND: Estimated life expectancy is one of the most important factors in determining treatment options for prostate cancer (PCa) patients. However, clinicians have few effective prognostic tools to individually assess survival in patients with PCa. METHODS: We screened 283,252 patients diagnosed with PCa from the Surveillance, Epidemiology, and End Results (SEER) database between 2004 and 2015, and randomly divided them into the training and validation groups. We used univariate and multivariate Cox analyses to identify independent prognostic factors and further established nomograms to predict 1-, 3-, 5-, and 10-year overall survival (OS) and cancer-specific survival (CSS) for PCa patients. The prediction performance of nomograms was tested and externally validated by Concordance index (C-index) and receiver operating characteristic (ROC) curve. Calibration curve and decision curve analysis (DCA) were used for internal validation. We further developed PCa prognostic scoring system based on the impact of available variables on survival. RESULTS: The variables age, race, marital status, TNM stage, surgery method, radiotherapy, chemotherapy, PSA value, and Gleason score identified as independent prognostic factors were included in the survival nomograms. The results of training (C-index: OS = 0.776, CSS = 0.889; AUC value: OS = 0.772-0.802, CSS = 0.892-0.936) and external validation (C-index: OS = 0.759, CSS = 0.875) indicated our nomograms had good performance in predicting 1-, 3-, 5-, and 10-year OS and CSS prediction. Internal validation using the calibration curves and DCA curves demonstrated the effectiveness of the prediction models. The prognostic scoring system was more effective than the AJCC staging system in predicting the survival of PCa patients, especially for OS. CONCLUSION: The prognostic nomograms and prognostic scoring system have favorable performance in predicting OS and CSS of PCa patients. These individualized survival prediction tools may contribute to clinical decisions.

A rare case of bladder cancer that metastasized to brain, heart, and lung lymph nodes benefited from immunotherapy.

Bladder cancer is a common malignant tumor of the genitourinary system, with the primary cause of death being metastasis. The most common metastatic sites are the lymph nodes, liver, lung, bone, peritoneum, pleura, kidney, adrenal gland, and the intestine. Brain and heart metastases are rare. In this report, we describe a patient who had pulmonary lymph node metastases more than a year after being diagnosed with bladder cancer, followed by brain and cardiac metastases more than two years later. Following the failure of standard first-line chemotherapy, the patient accepted 6 cycles of tislelizumab immunotherapy. The re-examination revealed that the bilateral frontal brain metastases had vanished, the right temporal lobe metastases had been greatly decreased, the neurological symptoms had been alleviated, and the cardiac metastases had disappeared. This is a rare clinical case with encouraging effects of tislelizumab and can serve as a model for the treatment of similar patients.

PD-1 engineered cytomembrane cloaked molybdenum nitride for synergistic photothermal and enhanced immunotherapy of breast cancer.

Incomplete tumor ablation and subsequent tumor metastasis usually occur during photothermal anti-tumor processes. The combination of photothermal and immunotherapy has proven to be a promising method to conquer technical challenges. Inhibiting the programmed death ligand-1 (PD-L1)/programmed cell death protein 1 (PD-1) immune pathway represents one of the most successful immunotherapy strategies. Whereas, the PD-L1 expression level significantly differs, leading to a relatively low response rate to the immune checkpoint blockade (ICB) approaches. Therefore, improving the expression level of PD-L1 becomes one potential method to enhance the response rate. Herein, NIH 3T3 cells were educated to steadily express PD-1 protein. Furthermore, the synthesized molybdenum nitride was then coated with PD-1 protein-modified cytomembrane, which endows it with immune checkpoint blocking capability. Moreover, under the irradiation of near-infrared light, the local mild heat released from the molybdenum nitride causes the apoptosis of tumor cells. More importantly, the elevated temperature simultaneously helps elevate the expression level of PD-L1, further enhancing the response rate of ICB. Finally, the PD-1 cytomembrane coatings interact with the upregulated PD-L1, leading to the activation of the immune system. In summary, we confirmed that the PD-1 protein-coated molybdenum nitride could synergistically ablate tumors and avoid metastasis.

Chiral aldehyde-nickel dual catalysis enables asymmetric α-propargylation of amino acids and stereodivergent synthesis of NP25302.

The combined catalytic systems derived from organocatalysts and transition metals exhibit powerful activation and stereoselective-control abilities in asymmetric catalysis. This work describes a highly efficient chiral aldehyde-nickel dual catalytic system and its application for the direct asymmetric α-propargylation reaction of amino acid esters with propargylic alcohol derivatives. Various structural diversity α,α-disubstituted non-proteinogenic α-amino acid esters are produced in good-to-excellent yields and enantioselectivities. Furthermore, a stereodivergent synthesis of natural product NP25302 is achieved, and a reasonable reaction mechanism is proposed to illustrate the observed stereoselectivity based on the results of control experiments, nonlinear effect investigation, and HRMS detection.

Screening and characterization estrogen receptor ligands from Arnebia euchroma (Royle) Johnst. via affinity ultrafiltration LC-MS and molecular docking.

Arnebiae Radix (dried root of Arnebia euchroma (Royle) Johnst.) is a traditional Chinese medicine (TCM) used to treat macular eruptions, measles, sore throat, carbuncles, burns, skin ulcers, and inflammations. The Arnebiae Radix extract can exert anti-breast cancer effects through various mechanisms of action. This study aimed to rapidly screen potential estrogen receptor (estrogen receptor α and estrogen receptor ß) ligands from the Arnebiae Radix extract. In this study, an analytical method based on affinity ultrafiltration coupled with UHPLC-Q-Exactive Orbitrap mass spectrometry was established for rapidly screening and identifying estrogen receptor ligands. Then, bindings of the components to the active site of estrogen receptor (estrogen receptor α and estrogen receptor ß) were investigated via molecular docking. Moreover, surface plasmon resonance (SPR) experiments with six compounds were performed to verify the affinity. As a result, a total of 21 ligands were screened from Arnebiae Radix using affinity ultrafiltration. Among them, 14 and 10 compounds from Arnebiae Radix showed affinity with estrogen receptor α and estrogen receptor ß, respectively. All of those ligands could have a good affinity for the multiple amino acid residues of the estrogen receptor based on molecular docking. In addition, six compounds display the great affinity by SPR. The method established in the study could be used to rapidly screen estrogen receptor ligands in Traditional Chinese medicine. The results demonstrated that the affinity ultrafiltration-UHPLC-Q-Exactive Orbitrap mass spectrometry method not only aids in the interpretation of the potential bioactive components and possible mechanisms of action of Arnebiae Radix but also provides a further effective basis for the quality control of this valuable herb medicine.

Prevention of Loperamide-Induced Constipation in Mice and Alteration of 5-Hydroxytryotamine Signaling by Ligilactobacillus salivarius Li01.

Although Ligilactobacillus salivarius Li01 (Li01) has shown much promise in preventing multiple gastrointestinal diseases, the potential of the probiotic in alleviating constipation and the related mechanisms remain unclear. In this study, the effects of Li01 were evaluated in a loperamide-induced constipation mouse model. The results demonstrated that Li01 intervention can relieve constipation symptoms by improving water content, quantity, and morphology of feces and act as an intestinal barrier structure protector. Furthermore, Li01 can modulate gut motility (gastrointestinal transit rate), the fluid transit-associated expression of aquaporins, and the serum parameters vasoactive intestinal peptide, substance P, and somatostatin. Constipation significantly increased the levels of 5-hydroxytryotamine (5-HT) in serum (p < 0.01) and decreased the levels in the intestine (p < 0.001). Due to its function of elevating the expression of tryptophan hydroxylase 1, this was reversed after Li01 treatment. Li01 also promoted the expression of 5-HT receptor 3 and 4, indicating that the 5-HT signaling pathway may play a critical role in the mechanism by which Li01 alleviate constipation symptoms. Additionally, Li01 significantly altered the gut microbiota composition by enhancing the ratio of Firmicutes/Bacteroidetes and increasing the abundance of Rikenellaceae_RC9 genera. Based on the above results, Li01 may have the potential to effectively alleviate constipation by regulating the 5-HT pathway and alteration of the gut microbiota.