High-throughput sequencing reveals Jatrorrhizine inhibits colorectal cancer growth by ferroptosis-related genes.

BACKGROUND: Colorectal cancer is a malignant tumor that poses a serious threat to human health. The main objective of this study is to investigate the mechanism by which Jatrorrhizine (JAT), a root extract from Stephania Epigaea Lo, exerts its anticancer effects in colorectal cancer. METHODS: We initially assessed the inhibitory properties of JAT on SW480 cells using MTT and cell scratch assays. Flow cytometry was employed to detect cell apoptosis. Differentially expressed genes were identified through high-throughput sequencing, and they were subjected to functional enrichment and signaling pathway analysis and PPI network construction. RT-qPCR was used to evaluate gene expression and identify critical differentially expressed genes. Finally, the function and role of differentially expressed genes produced by JAT-treated SW480 cells in colorectal cancer will be further analyzed using the TCGA database. RESULTS: Our study demonstrated that JAT exhibits inhibitory effects on SW480 cells at concentrations of 12.5µM, 25µM, 50µM, and 75µM without inducing cell apoptosis. Through high-throughput sequencing, we identified 244 differentially expressed genes. KEGG and GO analysis of high-throughput sequencing results showed that differentially expressed genes were significantly enriched in MAPK, Wnt, and P53 signaling pathways. Notably, JAT significantly altered the expression of genes associated with ferroptosis. Subsequent RT-qPCR showed that the expression of ferroptosis genes SLC2A3 and ASNS was significantly lower in JAT-treated SW480 cells than in the control group. Analysis by TCGA data also showed that ferroptosis genes SLC2A3 and ASNS were significantly highly expressed in COAD. The prognosis of SLC2A3 was significantly worse in COAD compared to the normal group. SLC2A3 may be a core target of JAT for the treatment of COAD. CONCLUSIONS: JAT can inhibit COAD growth by ferroptosis-related genes. And it is a potential natural substance for the treatment of COAD.

The extraction and identification of active components of the sex pheromones of Asian citrus psyllid, Diaphorina citri.

In this study, we extracted and identified the active components of the Asian citrus psyllid, Diaphorina citri sex pheromones to provide a basis for further development of sex attractants. Under laboratory conditions, mating activity in D. citri started 3 d after emergence, which peaked at 6-7 d, and mating activity had no obvious peak during the observed period 7:00-21:00 h. Additionally, D. citri males were attracted to the emanations from conspecific females, especially to the n-hexane extracts of the pheromone. A total of 17 compounds were identified from the n-hexane extracts of female and male D. citri by gas chromatography-mass spectrometer (GC-MS). Among them, 13 compounds were identified from the female D. citri n-hexane extracts, of which 7 (dichloromethane, acetic acid, toluene, butyl acetate, ethyl carbamoylacetate, α-pinene, and 1-nonanal) were not found in the male D. citri n-hexane extracts. In addition, a total of 33 compounds were identified from the solid phase microextraction (SPME) volatiles of the male and female D. citri adults. Among these, 17 compounds were identified from the female D. citri volatiles, of which 6 (cycloheptatriene, 5-methyl-2-phenylindole, 1-dodecanol, cis-11-hexadecena, dodecyl aldehyde, and nerylacetone) were not identified in the volatiles of the D. citri males. It was found that males were significantly attracted to 0.1-10 µL/mL acetic acid and 1-nonanal with the selection rates ranging from 62.04%-70.56% and 62.22%-67.22%, respectively. Therefore, the results of this study suggest that acetic acid and 1-nonanal might be the active compounds of the female D. citri sex pheromones.

Risk factors for inspiratory muscle weakness in coronary heart disease.

BACKGROUND: Patients with coronary heart disease (CHD) are susceptible to lung function problems caused by respiratory muscle weakness. Many CHD patients show complications of respiratory muscle weakness, but the risk factors remain unclear. OBJECTIVE: To explore the risk factors for inspiratory muscle weakness in CHD. METHODS: This study enrolled 249 patients with CHD who underwent maximal inspiratory pressure (MIP) measurement between April 2021 and March 2022.According to the percentage of MIP (MIP/Predicted normal value [PNV]), patients were divided into the inspiratory muscle weakness (IMW) (n = 149) (MIP/PNV<70%) and control groups (n = 100) (MIP/PNV≥70༅). Clinical information and MIP of the two groups were collected and analyzed. RESULTS: The incidence of IMW was 59.8% (n = 149). Age (P < 0.001); history of heart failure (P < 0.001), hypertension (P = 0.04), and peripheral artery disease (PAD) (P = 0.001); left ventricular end-systolic dimension (P = 0.035); presence of segmental motion abnormality of the ventricular wall (P = 0.030); and high density lipoprotein cholesterol (P = 0.001) and N-terminal brain natriuretic peptide (NT-proBNP) levels (P < 0.001) in the IMW group were significantly higher than those in the control group. The proportion of anatomic complete revascularization (P = 0.009), left ventricular ejection fraction (P = 0.010), and alanine transaminase (P = 0.014) and triglycerides levels (P = 0.014) in the IMW group were significantly lower than those in the control group. Logistic regression analysis showed that anatomic complete revascularization (OR=0.350, 95%CI 0.157-0.781) and NT-proBNP level (OR=1.002, 95%CI 1.000-1.004) were independent risk factors for IMW. CONCLUSION: The independent risk factors for decreased IMW in patients with CAD were anatomic incomplete revascularization and NT-proBNP level.

High-Throughput Sequencing Reveals That Rotundine Inhibits Colorectal Cancer by Regulating Prognosis-Related Genes.

BACKGROUND: Rotundine is an herbal medicine with anti-cancer effects. However, little is known about the anti-cancer effect of rotundine on colorectal cancer. Therefore, our study aimed to investigate the specific molecular mechanism of rotundine inhibition of colorectal cancer. METHODS: MTT and cell scratch assay were performed to investigate the effects of rotundine on the viability, migration, and invasion ability of SW480 cells. Changes in cell apoptosis were analyzed by flow cytometry. DEGs were detected by high-throughput sequencing after the action of rotundine on SW480 cells, and the DEGs were subjected to function enrichment analysis. Bioinformatics analyses were performed to screen out prognosis-related DEGs of COAD. Followed by enrichment analysis of prognosis-related DEGs. Furthermore, prognostic models were constructed, including ROC analysis, risk curve analysis, PCA and t-SNE, Nomo analysis, and Kaplan-Meier prognostic analysis. RESULTS: In this study, we showed that rotundine concentrations of 50 µM, 100 µM, 150 µM, and 200 µM inhibited the proliferation, migration, and invasion of SW480 cells in a time- and concentration-dependent manner. Rotundine does not induce SW480 cell apoptosis. Compared to the control group, high-throughput results showed that there were 385 DEGs in the SW480 group. And DEGs were associated with the Hippo signaling pathway. In addition, 16 of the DEGs were significantly associated with poorer prognosis in COAD, with MEF2B, CCDC187, PSD2, RGS16, PLXDC1, HELB, ASIC3, PLCH2, IGF2BP3, CLHC1, DNHD1, SACS, H1-4, ANKRD36, and ZNF117 being highly expressed in COAD and ARV1 being lowly expressed. Prognosis-related DEGs were mainly enriched in cancer-related pathways and biological functions, such as inositol phosphate metabolism, enterobactin transmembrane transporter activity, and enterobactin transport. Prognostic modeling also showed that these 16 DEGs could be used as predictors of overall survival prognosis in COAD patients. CONCLUSIONS: Rotundine inhibits the development and progression of colorectal cancer by regulating the expression of these prognosis-related genes. Our findings could further provide new directions for the treatment of colorectal cancer.

In vitro antifungal activity of lasiodiplodin, isolated from endophytic fungus Lasiodiplodia pseudotheobromae J-10 associated with Sarcandra glabra and optimization of culture conditions for lasiodiplodin production.

A macrolide antibiotic, lasiodiplodin was isolated from the endophytic fungus (EF) Lasiodiplodia pseudotheobromae J-10 associated with the medicinal plant Sarcandra glabra. In vitro antifungal assay demonstrated the inhibitory activity of lasiodiplodin against the growth of six phytopathogenic fungi, with the IC50 values ranging between 15.50 and 52.30 µg/mL. The highest antifungal activities were recorded against Exserohilum turcicum, Colletotrichum capsici, and Pestalotiopsis theae, with IC50 values of 15.50, 15.90, and 17.55 µg/mL, respectively. The underlying mechanism of the antifungal activity of lasiodiplodin against E. turcicum included the alteration of its colony morphology and disturbance of its cell membrane integrity. In addition, the optimization of L. pseudotheobromae J-10 culture conditions increased lasiodiplodin yield to 52.33 mg/L from 0.59 mg/L at pre-optimization. This is the first report on the isolation and identification of antifungal compound from the EF L. pseudotheobromae J-10 associated with S. glabra, as well as on the optimization of L. pseudotheobromae J-10 culture conditions to increase lasiodiplodin yield. The results of this study support that lasiodiplodin is a natural compound with high potential bioactivity against phytopathogens, and provide a basis for further study of the EF associated with S. glabra.

miR-16-5p Regulates Ferroptosis by Targeting SLC7A11 in Adriamycin-Induced Ferroptosis in Cardiomyocytes.

Introduction: Adriamycin (ADR) is commonly used in tumor chemotherapy, but its nonreversible cardiotoxicity severely hampers its clinical application. Ferroptosis is an implicated cause of ADR-induced injury. However, the underlying molecular mechanisms remain poorly understood. This study explored whether ferroptosis is a pivotal pathogenic pathway underlying ADR-induced cardiotoxicity and the possible molecular mechanisms involved. Methods: In vivo and in vitro experimental models were used to study the mechanism of ADR-mediated ferroptosis. Ferroptosis levels were examined in mice and human/rat cardiomyocytes. Mechanistically, the expression levels of SLC7A11 and related miRNAs were examined. Bioinformatics prediction and luciferase reporter assays were used to verify the potential interaction between miR-16-5p and SLC7A11. The biological functions and interaction of SLC7A11 and miR-16-5p were investigated in vivo and in vitro. Results: Our study observed that ADR treatment significantly increased ferroptosis levels in vivo and in vitro. Ferroptosis-related pharmacological interventions further confirmed these results. Our data displayed that the SLC7A11 level was significantly decreased in cardiac tissues and cells, while an increased expression level of miR-16-5p was observed. Moreover, upregulation of SLC7A1 and inhibition of miR-16-5p attenuated ADR-induced cardiomyocyte ferroptosis injury. Interactive rescue experiments showed that the protective effects of miR-16-5p inhibition on ADR-induced cardiomyocyte injury were blocked by SLC7A11 knockdown. Discussion: Based on these findings, targeting miR-16-5p could partially reverse the ADR-induced cardiotoxicity by rescuing the SLC7A11 to attenuate ferroptosis. This study presents a pre-clinical basis to identify miR-16-5p/SLC7A11 as a potential treatment target for ADR-induced cardiotoxicity.

Zinc finger C3H1-type containing serves as a novel prognostic biomarker in human pan-cancer.

BACKGROUND: Zinc finger C3H1 domain-containing protein (ZFC3H1) is differentially expressed between primary tumor and the normal in most cancers. Additionally, a recent study has suggested that ZFC3H1 could serve as a novel marker for the prognosis of prostate adenocarcinoma (PRAD). However, the relationship between ZFC3H1 expression and the prognostic values in most tumors remains unclear. Our study is mainly for exploring the prognosis of ZFC3H1 in pan-cancer and for further discovering a potential therapeutics target. METHODS: Based on the clinical big data, we performed a pan-cancer analysis of ZFC3H1, including gene expression, survival prognosis, genetic alteration, protein phosphorylation, immune infiltration and enrichment analysis. In addition, Real-Time PCR and Western Blot were used to further confirm the role of ZFC3H1 in the colorectal cancer. RESULTS: We found that ZFC3H1 expression was connected with the prognosis of multiple malignant tumors. Furthermore, we also observed that ZFC3H1 was highly expressed in colorectal cancer through Real-Time PCR and Western Blot. The primary tumors presented higher phosphorylation level of the S655 site in lung adenocarcinoma, colon adenocarcinoma and uterine corpus endometrial carcinoma. ZFC3H1 expression was positively correlated with the immune infiltration of Cancer-associated fibroblasts (CAFs) in some tumors, such as liver hepatocellular carcinoma. And RNA surveillance pathways may be closely associated with the occurrence of tumors. CONCLUSIONS: Our study first reveals that ZFC3H1 could serve as a novel prognostic biomarker of pan-cancer, especially colorectal cancer.

FAM65A as a novel prognostic biomarker in human tumors reveal by a pan-cancer analysis.

BACKGROUND: Family with sequence similarity 65 member A (FAM65A), also known as RIPOR1, is differentially expressed between human tumor and non-tumor tissues in kinds of cancers. In addition, it was reported that the product of FAM65A may be a biomarker for cholangiocarcinoma patients. However, there is still no evidence on the relationship between the FAM65A and different types of tumors. Our study is mainly for exploring the prognostic values of FAM65A in pan-cancer and for further discovering a potential therapeutics target. METHODS: We analyzed FAM65A expression, prognostic values, genetic alteration, protein phosphorylation, immune infiltration and enrichment analysis across different types of human malignant tumors based on The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) datasets. Additionally, Real-Time PCR (RT-qPCR) was performed to further confirm the roles of FAM65A in the pathogenesis of colorectal cancer. RESULTS: We found that FAM65A expression was associated with the prognosis of multiple human tumors, especially colorectal cancer. Moreover, we also observed that FAM65A was highly expressed in colorectal cancer through RT-qPCR. We observed that decreasing phosphorylation level of the S351 locus in colon adenocarcinoma, uterine corpus endometrial carcinoma and lung adenocarcinoma. And the expression of FAM65A was positively related to cancer-associated fibroblasts (CAFs) infiltration in many tumors, such as colon adenocarcinoma. Therefore, FAM65A may be a potential prognostic biomarker of human tumors.

Acaricidal activity against Panonychus citri and active ingredient of the mangrove plant Cerbera manghas.

Cerbera manghas is a mangrove plant which possesses comprehensive biological activities. A great deal of research has been undertaken on the chemical constituents and medical functions of C. manghas; insecticidal and antifungal activities have also been reported, but the acaricidal activity has not been studied. In our study, the acaricidal activity and active substances of C. manghas were investigated using a spray method, which showed that the methanol extracts of the fruit, twigs and leaves exhibited contact activity against female adults of Panonychus citri, with LC50 values at 24 h of 3.39 g L(-1), 4.09 g L(-1) and 4.11 g L(-1), respectively. An acaricidal compound was isolated from C. manghas by an activity-guided isolation method, and identified as (-)-17ß-neriifolin, which is a cardiac glycoside. (-)-17ß-Neriifolin revealed high contact activity against female adults, nymphae, larvae and eggs of P. citri, with LC50 values at 24 h of 0.28 g L(-1), 0.29 g L(-1), 0.28 g L(-1) and 1.45 g L(-1), respectively.

[Resonance scattering spectral assay of papain enzymatic activity with sodium dodecyl benzene sulfonate].

In pH 6.5 phosphate buffer solutions, dodecyl benzene sulfonate (SDBS) was combined with casein to form association particles, which exhibited five Rayleigh scattering peaks at 470, 360, 400, 420 and 520 nm, respectively. Under suitable conditions, papain has catalytic effect on the hydrolysis of casein, and SDBS can stop the catalytic reaction and be combined with the excess casein to form association particles. The scattering peak at 470 nm decreased with the activity of papain. The delta I470 value was linear with the papain activity in the range of 0.048-4.8 USP x mL(-1). Its regress equation is delta I(SDBS) = 1.972c + 2.31, with a related coefficient of 0.9999 and detection limit of 0.020 USP x mL(-1). This new assay has been applied to the assay of the papain activity in food additive with satisfactory results.