Toonanoronoids A-E, five new limonoids from Toona ciliata var. yunnanensis.

Five new B-seco-limonoids, namely toonanoronoids A-E (1-5), in conjunction with three previously reported compounds, were isolated from the EtOAc extract of the twigs and leaves of Toona ciliata var. yunnanensis. Their structures were elucidated through comprehensive spectroscopic and X-ray crystallographic analysis. The cytotoxic activities of new compounds against five human tumor cell lines (HL-60, SMMC-7721, A549, MCF-7, and SW480) were screened, Compounds 4 and 5 exerted inhibition toward two tumor cell lines (HL-60, SW-480) with IC50 values between 1.7 and 5.9 µM.

Histone lactylation inhibits RARγ expression in macrophages to promote colorectal tumorigenesis through activation of TRAF6-IL-6-STAT3 signaling.

Macrophages are phenotypically and functionally diverse in the tumor microenvironment (TME). However, how to remodel macrophages with a protumor phenotype and how to manipulate them for therapeutic purposes remain to be explored. Here, we show that in the TME, RARγ is downregulated in macrophages, and its expression correlates with poor prognosis in patients with colorectal cancer (CRC). In macrophages, RARγ interacts with tumor necrosis factor receptor-associated factor 6 (TRAF6), which prevents TRAF6 oligomerization and autoubiquitination, leading to inhibition of nuclear factor κB signaling. However, tumor-derived lactate fuels H3K18 lactylation to prohibit RARγ gene transcription in macrophages, consequently enhancing interleukin-6 (IL-6) levels in the TME and endowing macrophages with tumor-promoting functions via activation of signal transducer and activator of transcription 3 (STAT3) signaling in CRC cells. We identified that nordihydroguaiaretic acid (NDGA) exerts effective antitumor action by directly binding to RARγ to inhibit TRAF6-IL-6-STAT3 signaling. This study unravels lactate-driven macrophage function remodeling by inhibition of RARγ expression and highlights NDGA as a candidate compound for treating CRC.

[Equivalence of combined decoction and mixed single decoctions of Gegen Qinlian Decoction in alleviating chemotherapy-associated diarrhea].

This study compared the chemical profiles, component content, dry paste yield, and pharmacological effects of samples obtained from the mixed single decoctions and the combined decoction of Gegen Qinlian Decoction(GQD), aiming to provide an experimental foundation for evaluating the equivalence of the two decocting methods and the suitability of TCM formula granules in clinical application. The same decoction process was used to prepare the combined decoction and mixed single decoctions of GQD. Ultra-performance liquid chromatography coupled with Q-Exactive Orbitrap mass spectrometry(UPLC-Q-Exactive Orbitrap MS) was employed to compare the chemical profiles between the two groups. High-performance liquid chromatography(HPLC) was used to compare the content of nine characteristic components between the two groups. Then, a delayed diarrhea mouse model induced by irinotecan was established to compare the pharmacological effects of the two groups on chemotherapy-induced diarrhea. The UPLC-Q-Exactive Orbitrap MS in ESI~+ and ESI~- modes identified 59 chemical components in the compound decoction and mixed single decoctions, which showed no obvious differences in component species. The content of baicalin and wogonoside was higher in the compound decoction, while that of puerarin, daidzein-8-C-apiosylglucoside, berberine, epiberberine, wogonin, glycyrrhizic acid, and daidzein was higher in the mixed single decoctions. Further statistical analysis revealed no significant difference in the content of the nine characteristic components between the compound decoction and the mixed single decoctions. The dry paste yield had no significant difference between the two groups. Compared with the model group, both compound decoction and mixed single decoctions alleviated the weight loss and reduced diarrhea index in mice. Both of them lowered the levels of tumor necrosis factor-α(TNF-α), interleukin-1ß(IL-1ß), cyclooxygenase-2(COX-2), intercellular adhesion molecule-1(ICAM-1), interleukin-10(IL-10), malondialdehyde(MDA), and nitric oxide(NO) in the colon tissue. Furthermore, they significantly increased the levels of glutathione peroxidase(GSH-Px) and superoxide dismutase(SOD). Hematoxylin-eosin(HE) staining showed that colon tissue cells were tightly arranged with clear nuclei in both groups without obvious difference. The compound decoction and mixed single decoctions showed no significant differences in chemical component species, content of nine characteristic components, dry paste yield, or the pharmacological effects on alleviating chemotherapy-induced diarrhea. The findings provide a reference for evaluating the flexibility and superiority of combined or single decocting method in the preparation of TCM decoctions or formula granules.

TRIM21 is a druggable target for the treatment of metastatic colorectal cancer through ubiquitination and activation of MST2.

Metastatic colorectal cancer (mCRC) is characterized by poorer prognosis of patients and limited therapeutic approach, partly due to the lack of effective target. Using mouse models and tumor organoids, this study reported a tripartite motif 21 (TRIM21) protein, exerting potential inhibitory effects on the invasion and metastasis of CRC. Mechanistically, TRIM21 directly interacted with and ubiquitinated MST2 at lysine 473 (K473) via K63-linkage. This ubiquitination enabled the formation of MST2 homodimer and enhanced its kinase activity, ultimately resulting in the functional inactivation of yes-associated protein (YAP) and inhibition of an epithelial-mesenchymal transition (EMT) feature. We identified that vilazodone, an antidepressant, directly bound to TRIM21 to exert effective anti-metastatic action both in vitro and in vivo. Collectively, these findings revealed a previously unrecognized interplay between TRIM21 and the Hippo-YAP signaling. These results suggested that vilazodone could be repositioned as an anti-tumor drug to inhibit CRC metastasis by targeting TRIM21.

Two-Dimensional Nitrogen-Doped Carbon Nanosheets Derived from g-C3N4 /ZIF-8 for Solid-Phase Microextraction in Exhalation of Esophageal Cancer Patients.

Here, two-dimensional (2D) nitrogen-doped carbon nanosheets (CNSs) were prepared through carbonizing MOFs (ZIF-8) in-situ grown using graphitic carbon nitride (g-C3N4) as a template. The developed ZIF-8 CNS was then used as solid-phase microextraction (SPME) fiber coating for beneficiation of five biomarkers in exhalation of patients with esophageal cancer and in gas chromatography-mass spectrometry (GC-MS) for determination. The ZIF-8 CNS fiber exhibits satisfactory enrichment factors (3490-5631), wide linearity (5-1000 µg L-1), and low detection limits (0.26-0.96 µg L-1). The relative standard deviations (RSDs) for six replicate extractions of the same ZIF-8 CNS fiber were between 2.0-3.9% (intra-day) and 2.8-5.2% (inter-day). The reproducibility of three fibers prepared by the same approach was in the range 6.8-12.3% (RSD). The developed ZIF-8 CNS fiber can persist in 120 SPME cycles with no prominent loss of extraction efficiency and precision. The high enrichment factors of the 2D ZIF-8 CNS coatings are attributed to the high specific surface area, ultrathin thickness, and nano-pore or interlayer channels; moreover, nitrogen doping also endows the π system with a strong electron absorption ability, which will enhance the π-π interaction between the ZIF-8 CNS and the aromatic ring. Ultimately, the self-made ZIF-8 CNS-coated SPME fiber was applied to the analysis of exhaled breath samples. The recoveries of spiked analytes are between 84 and 105%.

Methylprednisolone for idiopathic granulomatous mastitis: a prospective observational cohort study.

Background: Idiopathic granulomatous mastitis (IGM) is a rare, benign, but locally aggressive breast disease. Steroids are widely used as a breast-conserving option, however, the response rate of steroids varies in reported studies, as well as its different reported usage. This prospective observational cohort study aimed to report the outcomes of methylprednisolone for IGM treatment. Methods: From Aug 2019 to Dec 2021, the clinicopathological information of 156 IGM patients who sought treatment at West China Hospital was prospectively collected. A total of 88 patients treated with methylprednisolone were included in the study. The clinical features, treatment response, and follow-up data were analyzed. Results: The median age at diagnosis was 32 years, and 90.9% of patients were multipara. The predominant symptom at presentation was painful breast mass, with a median size of 4.7 cm. For steroid usage, an initial 20 mg methylprednisolone daily was given until disease stable. The median duration of 20 mg methylprednisolone treatment was 45 (range, 14-376) days. The median duration of whole steroid therapy was 105 (range, 28-381) days. A total of 80.7% of patients (71/88) responded well to steroid treatment. In 63 patients, steroid treatment was successfully withdrawn, and treatment was completed. With an average of 283 days follow-up (range, 0-770 days), relapse was observed in 21 (33.33%) patients. Compared with patients with residual disease as shown by physical examination (PE), those with complete clinical remission (CCR) at the end of treatment had longer relapse-free intervals. Conclusions: Steroids are the preferable breast-conserving option for IGM. Treatment with 20 mg methylprednisolone for an average of 1.5 months is usually required, and full steroid treatment might last for 3 months.

Oncogenic potential of BEST4 in colorectal cancer via activation of PI3K/Akt signaling.

BEST4 is a member of the bestrophin protein family that plays a critical role in human intestinal epithelial cells. However, its role and mechanism in colorectal cancer (CRC) remain largely elusive. Here, we investigated the role and clinical significance of BEST4 in CRC. Our results demonstrate that BEST4 expression is upregulated in clinical CRC samples and its high-level expression correlates with advanced TNM (tumor, lymph nodes, distant metastasis) stage, LNM (lymph node metastasis), and poor survival. Functional studies revealed that ectopic expression of BEST4 promoted CRC cell proliferation and metastasis, whereas the depletion of BEST4 had the opposite effect both in vitro and in vivo. Mechanistically, BEST4 binds to the p85α regulatory subunit of phosphatidylinositol-3-kinase (PI3K) and promotes p110 kinase activity; this leads to activation of Akt signaling and expression of MYC and CCND1, which are critical regulators of cell proliferation and metastasis. In clinical samples, the expression of BEST4 is positively associated with the expression of phosphorylated Akt, MYC and CCND1. Pharmacological inhibition of Akt activity markedly repressed BEST4-mediated Akt signaling and proliferation and metastasis of CRC cells. Importantly, the interaction between BEST4 and p85α was also enhanced by epidermal growth factor (EGF) in CRC cells. Therapeutically, BEST4 suppression effectively sensitized CRC cells to gefitinib treatment in vivo. Taken together, our findings indicate the oncogenic potential of BEST4 in colorectal carcinogenesis and metastasis by modulating BEST4/PI3K/Akt signaling, highlighting a potential strategy for CRC therapy.

Transcriptomic Analysis Identifies Complement Component 3 as a Potential Predictive Biomarker for Chemotherapy Resistance in Colorectal Cancer.

Objective: 5-fluorouracil- and oxaliplatin-based FOLFOX regimens are mainstay chemotherapeutics for colorectal cancer (CRC) but drug resistance represents a major therapeutic challenge. To improve patient survival, there is a need to identify resistance genes to better understand the mechanisms underlying chemotherapy resistance. Methods: Transcriptomic datasets were retrieved from the Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases and combined with our own microarray data. Weighted gene co-expression network analysis (WGCNA) was used to dissect the functional networks and hub genes associated with FOLFOX resistance and cancer recurrence. We then conducted analysis of prognosis, profiling of tumor infiltrating immune cells, and pathway overrepresentation analysis to comprehensively elucidate the biological impact of the identified hub gene in CRC. Results: WGCNA analysis identified the complement component 3 (C3) gene as the only hub gene associated with both FOLFOX chemotherapy resistance and CRC recurrence after FOLFOX chemotherapy. Subsequent survival analysis confirmed that high C3 expression confers poor progression-free survival, disease-free survival, and recurrence-free survival. Further correlational analysis revealed significant negative association of C3 expression with sensitivity to oxaliplatin, but not 5-fluorouracil. Moreover, in silico analysis of tumor immune cell infiltration suggested the change of C3 expression could affect tumor microenvironment. Finally, gene set enrichment analysis (GSEA) revealed a hyperactivation of pathways contributing to invasion, metastasis, lymph node spread, and oxaliplatin resistance in CRC samples with C3 overexpression. Conclusion: Our results suggest that high C3 expression is a debilitating factor for FOLFOX chemotherapy, especially for oxaliplatin sensitivity, and C3 may represent a novel biomarker for treatment decision of CRC.

C21, C22 pregnane glycosides and cytotoxic C27 spriostanol steroids from Asparagus cochinchinesis.

A preliminary chemical investigation on 70% MeOH extract of the roots of Asparagus cochinchinensis resulted in the isolation of nine steroids. These isolates comprised of four new C21 (1-4) and one new pregnane (5) glycosides, and four known C27 (6-9) spirostanol steroids. Their structures were identified via analysis of the spectroscopic data and the results of hydrolytic cleavage. The cytotoxic activities of the compounds were tested toward the human tumor cell line Hela (cervical cancer), and compounds 7 and 8 displayed moderate activity with IC50 values of 35.5 and 39.6 µM, respectively.

[Study on the correlation factors on CD44+ CD24-/low ESA+ Lin- cell ratio in breast cancer].

OBJECTIVE: To explore the related factors on CD44+ CD24-/low ESA+ Lin- cell (breast caner stem cell) ratio in breast cancer patients. METHODS: Flow cytometry was used to analyze CD44+ CD24-/Iow ESA+ Lin- cell ratio in fresh tumor tissues of 40 breast cancer patients administered between March of 2011 and January of 2012 in West China Hospital, Sichuan University, China. Nine individual variables, such as age, clinical stage, histological grade, axillary lymph nodes metastases, immunohistochemisty (IHC) of estrogen receptor (ER), progesterone receptor (PR), C-erbB-2, and Ki-67 were selected to study their correlation with CD44+ CD24-/low ESA+ Lin- cell ratio. RESULTS: CD44+ CD24-/low ESA+ Lin- cells were expressed in all breast cancer tissues (median ratio: 4.38%, ranging from 0.5% to 16.8%), in which 20 patients (50.0%) showed low expression less than or equal to 4.38%, whereas, 20 patients (50.0%) demonstrated high expression greater than 4.38%. By univariant analysis, no statistical correlation was observed between CD44+ CD24-/low ESA+ Lin- cell ratio and the factors like age, tumor size, clinical stage, histological stage, axillary nodes metastasis metastasic, ER, PR. However, when Ki-67 was divided into high expression group (> 32.0%) and low expression group (< or = 32.0%), statistical difference in CD44+ CD24-/Iow ESA+ Lin- cell ratio between the two groups was observed (P = 0.008). By multivariate logistic analysis, only Ki-67 expression was the independent factor of breast cancer stem cell expression (P = 0.012, OR = 6.926, (95% CI, 1.529-31.377). CONCLUSION: In fresh breast cancer tissues, Ki-67 expression was the independent impact factor on CD44+ CD24-/low ESA+ Lin- cell ratio. Further investigation should be made to explore the correlation of breast cancer stem cell ratio with therapeutic effect and prognosis.

Constitutive and induced activities of defense-related enzymes in aphid-resistant and aphid-susceptible cultivars of wheat.

Phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO), and peroxidase (POD) are considered important biochemical markers in host plant resistance against pest insects. Constitutive activity of these enzymes was analyzed in resistant and susceptible wheat cultivars against cereal aphid Sitobion avenae (F.) at various developmental stages, i.e., tillering, stem elongation, flag leaf, and ear. Following aphid infestation, the activity of these enzymes was determined at the flag leaf and ear stages. Resistant cultivars exhibited greater constitutive PAL activity than susceptible ones at the tillering, stem elongation, and flag leaf stages. Aphid infestation enhanced levels of PAL activity in the flag leaf and ear stages in both resistant and susceptible cultivars. Constitutive PPO activity was higher in the resistant cultivars at all developmental stages. Aphid infestation induced increases in PPO activity in the flag leaf and ear stages of one susceptible cultivar, whereas induction in resistant cultivars was weaker. Resistant cultivars showed greater constitutive POD activity in the tillering, stem elongation, and flag leaf stages, while aphid infestation induced POD activity in all cultivars, especially in susceptible ones. The potential role of PAL, PPO, and POD in wheat defense against aphid infestation is discussed.