Oxypeucedanin hydrate alleviates rheumatoid arthritis by inhibiting the TLR4-MD2/NF-κB/MAPK signaling axis.

Rheumatoid arthritis (RA) is an idiopathic and chronic autoimmune disease for which there are currently no effective treatments. Oxypeucedanin hydrate (OXH) is a natural coumarin known for its potent anti-inflammatory properties. However, further investigations are needed to determine its therapeutic efficacy in treating RA. In this study, we evaluate the anti-inflammatory activity of OXH by treating LPS-induced RAW264.7 macrophages. Our results show that OXH treatment reverses the changes in iNOS, COX-2, IL-1ß, IL-6, and TNF-α levels. Additionally, OXH reduces ROS production. Further analysis reveals that OXH suppresses the activation of the NF-κB/MAPK pathway. CETSA results show that OXH competes with LPS for binding to the TLR4/MD2 complex. MST experiments demonstrate the specific affinity of OXH for the TLR4/MD2 complex, with a Kd value of 33.7 µM. Molecular docking analysis suggests that OXH binds to the pocket of the TLR4/MD2 complex and interacts with specific amino acids, such as GLY-343, LYS-388, and PHE-345. Molecular dynamics simulations further confirm this conclusion. Finally, we investigate the potential of OXH in treating RA using a collagen-induced arthritis (CIA) model in rats. OXH effectively ameliorates the symptoms of CIA, including improving body weight, reducing swelling and redness, increasing talus volume, and decreasing bone erosion. OXH also decreases the mRNA levels of pro-inflammatory factors in synovial tissue. Transcriptome enrichment analysis and western blot analysis confirm that OXH suppresses the NF-κB/MAPK pathway, which is consistent with our in vitro findings.

Polar Effects Control the Gas-Phase Reactivity of para-Benzyne Analogs.

We report herein a gas-phase reactivity study on a para-benzyne cation and its three cyano-substituted, isomeric derivatives performed using a dual-linear quadrupole ion trap mass spectrometer. All four biradicals were found to undergo primary and secondary radical reactions analogous to those observed for the related monoradicals, indicating the presence of two reactive radical sites. The reactivity of all biradicals is substantially lower than that of the related monoradicals, as expected based on the singlet ground states of the biradicals. The cyano-substituted biradicals show substantially greater reactivity than the analogous unsubstituted biradical. The greater reactivity is rationalized by the substantially greater (calculated) electron affinity of the radical sites of the cyano-substituted biradicals, which results in stabilization of their transition states through polar effects. This finding is in contrast to the long-standing thinking that the magnitude of the singlet-triplet splitting controls the reactivity of para-benzynes.

Chemical constitutes from Tuber indicum with immunosuppressive activity uncovered by transcriptome analysis.

Three previously undescribed compounds including a polyketide (1) and two lactams (2 and 3) were obtained from Tuber indicum. The structures of new findings were elucidated by HRESIMS, NMR as well as NMR and ECD calculations. Transcriptome analysis through RNA-seq revealed that compound 2 exhibits immunosuppressive activity. Lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages were employed as a model to explore the effect of these compounds in immunosuppressive activity. The results showed that 2 could reduce the generation of inflammatory mediators including nitric oxide (NO), reactive oxygen species (ROS), interleukin 6 (IL-6), tumor necrosis factor α (TNF-α), cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS). Western blotting analysis demonstrated that 2 could suppressed the PI3K pathway by decreasing the levels of p-PI3K and p-Akt, while increasing the levels of p-PTEN. The anti-inflammatory activity of 2 was further confirmed using a zebrafish in vivo model.

Differentiation of Seven Isomeric n-Pentylquinoline Radical Cations Based on Energy-Resolved Medium-Energy Collision-Activated Dissociation.

Asphaltenes, a major and undesirable component of heavy crude oil, contain many different types of large aromatic compounds. These compounds include nitrogen-containing heteroaromatic compounds that are thought to be the main culprit in the deactivation of catalysts in crude oil refinery processes. Unfortunately, prevention of this is challenging as the structures and properties of the nitrogen-containing heteroaromatic compounds are poorly understood. To facilitate their structural characterization, an approach based on ion-trap collision-activated dissociation (ITCAD) tandem mass spectrometry followed by energy-resolved medium-energy collision-activated dissociation (ER-MCAD) was developed for the differentiation of seven isomeric molecular radical cations of n-pentylquinoline. The fragmentation of each isomer was found to be distinctly different and depended largely on the site of the alkyl side chain in the quinoline ring. In order to better understand the observed fragmentation pathways, mechanisms for the formation of several fragment ions were delineated based on quantum chemical calculations. The fast benzylic α-bond cleavage that dominates the fragmentation of analogous nonheteroaromatic alkylbenzenes was only observed for the 3-isomer as the major pathway due to the lack of favorable low-energy rearrangement reactions. All the other isomeric ions underwent substantially lower-energy rearrangement reactions as their alkyl chains were found to interact mostly via 6-membered transition states either with the quinoline nitrogen (2- and 8-isomers) or the adjacent carbon atom in the quinoline core (4-, 5-, 6-, and 7-isomers), which lowered the activation energies of the fragmentation reactions. The presented analytical approach will facilitate the structural characterization of nitrogen-containing heteroaromatic compounds in asphaltenes.

Study of the Change Laws of Free Radicals and Functional Groups during Coal Oxidation.

In order to study the change laws of free radicals and functional groups during low-temperature coal oxidation, three coal samples with different metamorphic degrees were selected for ESR and FTIR analysis. The results showed that the concentration of free radicals increased as the temperature increased; meanwhile, the types of free radicals changed constantly, and the free radical variation range decreased with the increase in coal metamorphism. The side chains of aliphatic hydrocarbons in coal with a low metamorphic degree decreased by varying amounts in the initial heating stage. The -OH content of bituminous coal and lignite increased first and then decreased, while that in anthracite decreased first and then increased. In the initial oxidation stage, -COOH first increased rapidly, then decreased rapidly, and then increased before finally decreasing. The content of -C=O in bituminous coal and lignite increased in the initial stage of oxidation. Through gray relational analysis, it was found that there was a significant relationship between free radicals and functional groups, and -OH had the strongest correlation with free radicals. This paper provides a theoretical basis for studying the mechanism of functional groups transforming into free radicals in the process of coal spontaneous combustion.

Sanguisorba officinalis ethyl acetate extract attenuates ulcerative colitis through inhibiting PI3K-AKT/NF-κB/ STAT3 pathway uncovered by single-cell RNA sequencing.

BACKGROUND: Ulcerative colitis (UC) accounts for the untreatable illness nowadays. Bloody stools are the primary symptom of UC, and the first-line drugs used to treat UC are associated with several drawbacks and negative side effects. S. officinalis has long been used as a medicine to treat intestinal infections and bloody stools. However, what the precise molecular mechanism, the exact etiology, and the material basis of the disease remain unclear. PURPOSE: This work aimed to comprehensively explore pharmacological effects as well as molecular mechanisms underlying the active fraction of S. officinalis, and to produce a comprehensive and brand-new guideline map of its chemical base and mechanism of action. METHODS: First, different polarity S. officinalis extracts were orally administered to the DSS-induced UC model mice for the sake of investigating its active constituents. Using the UPLC-orbitrap high-resolution mass spectrometry (UPLC-Q-Orbitrap-HRMS) technique, the most active S. officinalis (S. officinalis ethyl acetate fraction, SOEA) extract was characterized. Subsequently, the effectiveness of its active fraction on UC was evaluated through phenotypic observation (such as weight loss, colon length, and stool characteristics), and histological examination of pathological injuries, mRNA and protein expression. Cell profile, cell-cell interactions and molecular mechanisms of SOEA in different cell types of the colon tissue from UC mice were described using single-cell RNA sequencing (scRNA-seq). As a final step, the molecular mechanisms were validated by appropriate molecular biological methods. RESULTS: For the first time, this study revealed the significant efficacy of SOEA in the treatment of UC. SOEA reduced DAI and body weight loss, recovered the colon length, and mitigated colonic pathological injuries along with mucosal barrier by promoting goblet cell proliferation. Following treatment with SOEA, inflammatory factors showed decreased mRNA and protein expression. SOEA restored the dynamic equilibrium of cell profile and cell-cell interactions in colon tissue. All of these results were attributed to the ability of SOEA to inhibit the PI3K-AKT/NF-κB/STATAT pathway. CONCLUSIONS: By integrating the chemical information of SOEA derived from UPLC-Q-Orbitrap-HRMS with single-cell transcriptomic data extracted from scRNA-seq, this study demonstrates that SOEA exerts the therapeutic effect through suppressing PI3K-AKT/NF-B/STAT3 pathway to improve clinical symptoms, inflammatory response, mucosal barrier, and intercellular interactions in UC, and effectively eliminates the interference of cellular heterogeneity.

Efficacy and safety of bipolar androgen therapy in mCRPC after progression on abiraterone or enzalutamide: A systematic review.

PURPOSE: To further determine the efficacy and safety of bipolar androgen therapy (BAT) on patients with metastatic castration-resistant prostate cancer (mCRPC) after progression on abiraterone (ABI) or enzalutamide (ENZA). MATERIALS AND METHODS: We systematically searched the Pubmed, Web of Science and ClinicalTrials.gov up to June 2021. Literature review, study selection, and data extraction were conducted by 2 reviewers. Risk of bias was assessed according to the methodology of the European Association of Urology (EAU). A systematic review and pooled analysis were performed. The primary outcomes were PSA50 after BAT and AR-targeted therapy rechallenge, objective response rate (ORR) after BAT, and AEs after BAT. The definition of PSA50 was that participants achieving a PSA decline ≥50% according to Prostate Cancer Working Group (PCWG2) criteria. The ORR determined by determined by Response Evaluation Criteria in Solid Tumors (RECIST) included patients experienced partial response (PR) or complete response (CR). RESULTS: In a total of 74 unique records, 5 studies were eligible for inclusion. Participants who underwent BAT achieved PSA50 of 0.26 (95% CI [0.20, 0.32]) and objective response rate (ORR) of 0.32 (95% CI [0.21, 0.44]). Patients completed BAT proceeded to AR-target therapy (ABI or ENZA) achieved moderate response (PSA50 0.54, 95% CI [0.30, 0.76]). Based on our multiple subgroup analysis, type of post-BAT AR-target therapy had a strong impact on PSA50 of AR-target therapy rechallenge. Most of adverse events (AEs) were low grade. CONCLUSIONS: The present study indicated that BAT could induce clinical responses in mCRPC patients after progression on ABI or ENZA, with an acceptable side effects profile. BAT could also be able to restore sensitivity to ABI and ENZA rechallenge in a subset of patients.

Combined Treatment of Radiotherapy and Immunotherapy for Urological Malignancies: Current Evidence and Clinical Considerations.

Although it has always been believed that radiation has immunosuppressive effects, more and more preclinical and clinical trials have shown that the combination of radiotherapy and immunotherapy has a potential synergistic effect to treat cancers including urological malignancies. When radiotherapy is combined with immunotherapy, improved prognosis has been observed in different urinary tumors. However, there is no standard treatment, such as the optimal dose/fractionation and the sequence of immunotherapy and radiotherapy. In this review, we discussed the effects of radiotherapy on the cancer immune system and emphasized the synergy of radiotherapy combined with immunotherapy. Although it has significantly improved the prognosis of tumors, there are still some unresolved questions about how to best use this combination in clinical practice. Ongoing trials will provide further information on the interaction of radiotherapy combined with immunotherapy, and are expected to guide clinical practice and improve clinical outcomes.

Association of Novel Androgen Receptor Axis-Targeted Therapies With Diarrhea in Patients With Prostate Cancer: A Bayesian Network Analysis.

OBJECTIVE: To perform a systematic review and network meta-analysis to characterize the effect of novel androgen receptor axis-target (ARAT) agents on diarrhea and constipation. METHODS: We searched the Pubmed, Web of Science, and ClinicalTrials.gov up to September 2021 for phase 3 randomized controlled trials (RCTs) of patients receiving novel ARAT agents for prostate cancer (CaP). A Cochrane risk-of-bias tool was used to assess trial quality. The primary outcomes were risk ratio (RR) of any-grade diarrhea and constipation for patients receiving ARAT treatment. RRs of competing treatments were evaluated by pairwise and Bayesian network meta-analysis. RESULTS: In this study, 13 trials with 15,117 participants comparing 5 treatments (abiraterone, enzalutamide, apalutamide, darolutamide, and placebo) were identified. Use of novel ARAT agents was associated with a significant increased risk of any-grade diarrhea (RR = 1.30, 95% CI [1.16, 1.44]). As for subgroup analysis, abiraterone, enzalutamide, and apalutamide were all associated with significant increased risk of any-grade diarrhea (abiraterone: RR = 1.40, 95% CI [1.09, 1.81]; enzalutamide: RR = 1.17, 95% CI [1.02, 1.35]; apalutamide: RR = 1.35, 95% CI [1.03, 1.76]). Based on Bayesian modeling, abiraterone and enzalutamide showed the highest and lowest probability to rank first in terms of increasing risk of any-grade diarrhea. There were no significant differences of risk in any-grade constipation, grade 3 or greater diarrhea, and constipation between ARAT and control group. CONCLUSION: The present study indicates that the use of novel ARAT agents is associated with a significantly higher risk of diarrhea. Across the four agents, abiraterone may relate to the highest risk of diarrhea among patients with metastatic hormone sensitive prostate cancer (mHSPC) and castration-resistant prostate cancer (CRPC).

Steroid switch after progression on abiraterone plus prednisone in patients with metastatic castration-resistant prostate cancer: A systematic review.

BACKGROUND: Emerging evidence indicates that patients with metastatic castration-resistant prostate cancer could respond to steroid switch from prednisone (P) to dexamethasone (D) following progression on abiraterone acetate plus prednisone (AA+P). OBJECTIVES: Conducting a systematic review to evaluate the efficacy, safety, and prognostic factors of steroid switch. MATERIALS AND METHODS: We systematically searched Pubmed, Web of Science, and American Society of Clinical Oncology annual meeting abstracts published up to October 2020. Literature review, study selection, and data extraction were conducted by two reviewers. Risk of bias (RoB) and quality of evidence were assessed. A systematic review and pooled analysis were performed. RESULTS: Nine studies were eligible for inclusion. All of the included patients were progression on AA+P. Pooled rates of PSA50 and PSA30 on abiraterone acetate plus dexamethasone (AA+D) were 0.24 (95%CI [0.18,0.30]) and 0.42 (95%CI [0.36,0.48]), respectively. Subgroup analysis indicated more favorable PSA50 and PSA30 rates on AA+D when switching from P to D only based on PSA progression. Median time to PSA progression on AA+D ranged from 2.73 to 11.38 months. Definitions of progression free survival were variable. Reported median progression free survival on AA+D ranged from 2.52 to 11.8 months. Median overall survival on AA+D varied from 4.11 to 20.9 months. All patients tolerated well on AA+D, and no grade 3 to 4 adverse events were reported. Baseline characteristics of patients, previous treatment and its response, and genetic alterations might all play roles in the response in the response toward the AA+D regimen. CONCLUSIONS: The present systematic review suggested that steroid switch from P to D might be an effective and safe treatment strategy in a subset of patients with metastatic castration-resistant prostate cancer after PSA progression on AA+P.

Clinical Characteristics and Prognostic Factors of Testicular Sarcoma: A Population-Based Study.

OBJECTIVES: To study clinical characteristics and factors that may affect the prognosis of testicular sarcoma patients. PATIENTS AND METHODS: In the Surveillance Epidemiology and End Results database (2006-2016), people with testicular sarcoma were enrolled in our research. Multivariable Cox proportional hazard model and Multivariable Logistic regression model were used to compare the impact of different factors on cancer-specific survival, localized metastasis, and distant metastasis. RESULTS: This research was based on the registry information of 158 testicular sarcoma patients. All patients with a median age of 17.00 (1.00-93.00) years were pathologically diagnosed with orchiectomy or needle biopsy specimens. Patients with Grade I, II, III, and IV testicular sarcoma accounted for 34.29% (n = 24), 10.10% (n = 7), 22.86% (n = 16), and 32.86% (n = 23) of all patients, respectively. There were 42 (30.43%), 53 (38.41%), 15 (10.87%), 20 (14.49%), 5 (3.62%), 3 (2.17%) patients with Tis, T1, T2, T3, T4, and >T4 (the invasion degree exceeded the staging system of testicular cancer) disease respectively. Among all included patients, localized metastasis occurred in 31 (20.13%) patients, distant metastasis was found in 28 (18.18%) patients during observation, and 61.69% (n = 95) had no metastasis. Thirty-two (20.25%) patients died of this cancer. According to our study, patients with distant metastasis [OR = 17.86, 95% CI (4.63-68.84), p < 0.0001] and T3 disease [OR = 4.13, 95% CI (1.10-15.53), p = 0.0359] were more likely to die of this cancer. Patients with advanced T stage were more likely to occur distant metastasis, [OR = 13.91, 95% CI (1.80-107.54), p = 0.0116] for T3 and [OR = 16.36, 95% CI (1.36-196.21), p = 0.0275] for T4. CONCLUSIONS: According to our research, factors including metastasis and higher T stage were significantly related with poorer prognosis of testicular sarcoma. Higher T stage was also found to be a risk factor of distant metastasis. The recognization of these poor prognostic factors may allow physicians to make comprehensive and appropriate management decision for testicular sarcoma patients.

Pretreatment elevated fibrinogen level predicts worse oncologic outcomes in upper tract urothelial carcinoma.

This study aimed to further validate the prognostic role of fibrinogen in upper tract urothelial carcinoma (UTUC) in a large Chinese cohort. A total of 703 patients who underwent radical nephroureterectomy were retrospectively identified. Fibrinogen levels of ≥4.025 g l-1 were defined as elevated. Logistic regression analysis was performed to determine the association between fibrinogen and adverse pathological features. Kaplan-Meier analysis and Cox regression models were used to assess the associations of fibrinogen with cancer-specific survival (CSS), disease recurrence-free survival (RFS), and overall survival (OS). Harrell c-index and decision curve analysis were used to assess the clinical utility of multivariate models. The median follow-up duration was 42 (range: 1-168) months. Logistic regression analysis revealed that elevated fibrinogen was associated with higher tumor stage and grade, lymph node involvement, lymphovascular invasion, sessile carcinoma, concomitant variant histology, and positive surgical margins (all P < 0.05). Multivariate Cox regression analysis demonstrated that elevated fibrinogen was independently associated with decreased CSS (hazard ratio [HR]: 2.33; P < 0.001), RFS (HR: 2.09; P < 0.001), and OS (HR: 2.09; P < 0.001). The predictive accuracies of the multivariate models were improved by 3.2%, 2.0%, and 2.8% for CSS, RFS, and OS, respectively, when fibrinogen was added. Decision curve analysis showed an added benefit for CSS prediction when fibrinogen was added to the model. Preoperative fibrinogen may be a strong independent predictor of worse oncologic outcomes in UTUC; therefore, it may be valuable to apply this marker to the current risk stratification in UTUC.

Metabolic syndrome and upper tract urothelial carcinoma: A retrospective analysis from a large Chinese cohort.

BACKGROUND: Metabolic syndrome (MetS) has been reported to be associated with adverse outcomes in cancer patients. However, the relationship between MetS and upper tract urothelial carcinoma (UTUC) has yet to be explored. OBJECTIVES: To investigate the prognostic value of MetS in UTUC after radical nephroureterectomy. PATIENTS AND METHODS: A total of 644 patients with UTUC after radical nephroureterectomy were identified at West China Hospital from May 2003 to December 2016. MetS was defined as the co-existence of 3 or more of 5 components (obesity, hypertension, elevated fasting glucose, decreased high-density lipoprotein-cholesterol, and hypertriglyceridemia). Logistic and Cox regression analyses were performed to evaluate the associations of MetS with pathological features and survival outcomes. Decision curve analysis and Harrell concordance index were used to determine the clinical utility of the prediction models. RESULTS: Of 644 patients, 157 (24.4%) had MetS. Over a median follow-up of 39 months, 269 (41.8%) experienced disease recurrence, 233 (36.2%) died, and 185 (28.7%) died of UTUC. MetS was independently associated with high-grade disease, advanced pT stage (≥pT3), and lymphovascular invasion (each P < 0.05). Multivariate Cox regression analysis showed that MetS was an independent factor for decreased cancer-specific survival (hazard ratio [HR]: 1.38, 95% confidence intervals [CI]: 1.01-1.89, P = 0.042) but not for recurrence-free survival (HR: 1.27, 95% CI: 0.97-1.67, P = 0.078), and overall survival (HR: 1.24, 95% CI: 0.95-1.62, P = 0.121). The estimated c-index of the multivariate models for cancer-specific survival was 0.763 compared with 0.769 when MetS added. CONCLUSIONS: MetS is a negative prognostic factor in UTUC. Further studies of MetS in UTUC are demanded.

Validation of the preoperative controlling nutritional status score as an independent predictor in a large Chinese cohort of patients with upper tract urothelial carcinoma.

BACKGROUND: Pretreatment controlling nutritional status (CONUT) score is a novel index which was used to predict outcomes in cancer patients. We aim to explore the prognostic significance of CONUT score in patients with upper tract urothelial carcinoma (UTUC) after radical nephroureterectomy (RNU). PATIENTS AND METHODS: A total of 662 UTUC patients between 2004 and 2016 were retrospectively analyzed. Patients were categorized into three groups based on CONUT score (Normal: 0-1; Light: 2-4; Moderate/severe: 5-12). Associations of CONUT score with oncological outcomes were analyzed using Logistic and Cox regression analysis. Harrell concordance index was used to assess the predictive accuracy of the multivariate models. Subgroup analyses were conducted according to tumor grade and stage. RESULTS: The median follow-up duration was 41 months. Multivariate Logistic analysis showed that high CONUT score was independently associated with high-grade disease, high pT stage, lymphovascular invasion, sessile carcinoma, variant histology, and positive surgical margins (each P < 0.05). Multivariate analysis demonstrated that CONUT score 5-12 was an independent factor for worse cancer-specific survival (CSS, hazard ratio [HR]:2.39, 95% confidence interval [CI] 1.55-3.68, P < 0.0001), disease recurrence-free-survival (RFS, HR: 1.80, 95% CI 1.24-2.60, P = 0.002), and overall survival (OS, HR: 2.26, 95% CI 1.53-3.34, P < 0.0001). The estimated c-index of the multivariate models for CSS, RFS, and OS increased from 0.755, 0.715 and 0.745 to 0.772, 0.723, and 0.756 when CONUT score supplemented. Subgroup analyses showed that especially in patients with high-grade carcinoma and advanced stage (≥pT3), higher CONUT score predicts decreased CSS, RFS, and OS (all P < 0.05). CONCLUSION: Preoperative CONUT score is a negative independent prognostic indicator for both pathologic and survival outcomes in UTUC, especially in those with high-grade carcinoma and advanced stage. Adding this parameter into our clinical prediction model is appropriate so as to improve its predictive accuracy.