Topiramate inhibits the proliferation of bladder cancer cells via PI3K/AKTR signaling pathway

Purpose: To explore new treatment options for bladder cancer (BC) based on topiramate (TPM).Methods: The MTT assay and flow cytometry were used to determine the effect of topiramate on partial growth-related malignant phenotype of BC cells. Expression levels of apoptosis-related biomarkers and signaling pathway-related factors were analyzed using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. In vivo experiments were conducted to investigate the role of TPM on tumor growth in mice with bladder cancer.Results: The MTT results showed that topiramate blocked the growth of BC cells (p < 0.05). Growth inhibition was positively correlated with TPM concentration. Flow cytometry results revealed that bladder cancer cell apoptosis rose with increase in TPM concentration, while the mRNAs of apoptosisassociated factors Bcl-2 and Mcl-1 were down-regulated in a concentration-based manner by TPM (p < 0.05). Western blot assay indicated that Bax and Caspase-3 proteins were up-regulated, and the higher the concentration of TPM, the more significant the protein expression levels (p < 0.05).Conclusion: Topiramate (TPM) slows down the rate of growth of BC cells and accelerates their rate of apoptosis through the regulation of P13K/AKT/mTOR signaling pathway. Thus, the compound has potentials for development as an anti-bladder cancer agent

Differential effects of domesticated and wild Capsicum frutescens L. on microbial community assembly and metabolic functions in rhizosphere soil

ObjectiveRhizosphere microorganisms play crucial roles in the growth and development of plants, disease resistance, and environmental adaptability. As the only wild pepper variety resource in China, domesticated Capsicum frutescens Linn. (Xiaomila) exhibits varying beneficial traits and affects rhizosphere microbial composition compared with its wild counterparts. In this study, we aimed to identify specific rhizosphere microbiome and metabolism patterns established during the domestication process.MethodsThe rhizosphere microbial diversity and composition of domesticated and wild C. frutescens were detected and analyzed by metagenomics. Non-targeted metabolomics were used to explore the differences of metabolites in rhizosphere soil between wild and domesticated C. frutescens.ResultsWe found that the rhizosphere microbial diversity of domesticated variety was significantly different from that of the wild variety, with Massilia being its dominant bacteria. However, the abundance of certain beneficial microbes such as Gemmatimonas, Streptomyces, Rambibacter, and Lysobacter decreased significantly. The main metabolites identified in the wild variety included serylthreonine, deoxyloganic acid, vitamin C, among others. In contrast, those identified in the domesticated group were 4-hydroxy-l-glutamic acid and benzoic acid. Furthermore, the differentially enriched pathways were concentrated in tyrosine and tryptophan biosynthesis, histidine and purine-derived alkaloids biosynthesis, benzoic acid family, two-component system, etc.ConclusionThis study revealed that C. frutescens established specific rhizosphere microbiota and metabolites during domestication, which has important significance for the efficient utilization of beneficial microorganisms in breeding and cultivation practices

Relationship Between Early Oral Intake Post Pancreaticoduodenectomy and Chyle Leakage: A Retrospective Cohort Study

Background Early oral intake is strongly recommended according to the enhanced recovery after surgery (ERAS) guidelines because it can reduce complications and improve recovery. However, early oral intake has been indicated to be associated with chyle leakage (CL) after pancreatic surgery, which may lead to worsening of existing malnutrition and impeded recovery. This study investigated the relationship between early oral intake and CL and identified risk factors for CL to reduce its occurrence and promote recovery after pancreaticoduodenectomy. Materials and Methods All patients who underwent pancreaticoduodenectomy between June 2014 and June 2018 were identified retrospectively. Patients were divided into the early-oral-intake and control groups according to whether they had early oral intake according to ERAS protocols. CL and other clinicopathological characteristics were recorded. Univariable and multivariable analyses assessed CL risk factors. Results Early oral intake improved recovery, leading to a shorter postoperative hospital stay for the early-oral-intake group in comparison to that of the control group [13.6 (range, 12–68) vs. 17.8 (range, 14–83) days; p = 0.047] without increasing the incidence of CL and other complications. CL was diagnosed significantly earlier in the early-oral-intake group than in the control group [4.6 (range 3–5) vs. 6.7 (range 3–9) days; p = 0.001]. Early oral intake did not increase the grade severity (p = 0.845) or the costs (p = 0.241) or prolong postoperative hospital stays (p = 0.611). A primary diagnosis of malignancy, para-aortic lymph node dissection, lymphatic invasion, lymph node metastases, the number of harvested nodes, and the number of positive nodes were significantly associated with CL (p < 0.05), whereas early oral intake was not (p = 0.525). Multivariate analyses demonstrated that para-aortic lymph node dissection (p = 0.039) and the number of harvested nodes (p = 0.001) were independent risk variables. Conclusion This study provides significant evidence that early oral intake after pancreaticoduodenectomy is not associated with CL. The identification of the independent risk factors for CL can help prevent it

Representative spectrogram of an 18-week-old BKS.Cg db/db mouse (B18-1).

The x-axis indicates time (s), and the y-axis indicates frequency (Hz). The conditions for STFT were as follows: FFT points, 1024; separate points, 512; shift length, 32; window length, 64.</p

Graphical flow chart from the animal experiments to the multivariate analysis.

In this figure, the results of NMR analysis of blood samples collected after animal experiments in groups A (n = 10) and B (n = 10) are shown. In total, 20 1H-NMR FID signal data were acquired, which were short-time Fourier transformed [51], and 20 spectrograms were calculated. For multivariate analysis, a process was performed to obtain a data matrix from the 10 spectrograms. From this data matrix, an exploratory analysis was performed using principal component analysis [26] for the identification of outliers and potential subgroups of animals in groups A and B. From this figure, it was possible to identify subgroups that were not initially anticipated, so PLS-DA [52, 53] was performed only with data that corresponded to subgroups in the same data matrix to analyze whether subgroups could be identified. In both principal component analysis and PLS-DA, the variables that were important for the identification of groups in each analysis were plotted as corresponding score plots. This score plot has the same time and frequency axes as the spectrogram and thus can be compared with the spectrogram. The illustration of the mouse and the NMR equipment in this figure was obtained from Research Net, a website where research illustration material is available free of charge (https://www.wdb.com/kenq/illust © WDB Co., Ltd., Tokyo, Japan). References (cited only in the Supporting Information) 51. Wacker M, Witte H. Time-frequency techniques in biomedical signal analysis. a tutorial review of similarities and differences. Methods Inf Med. 2013;52:279–296. 52. Barker M, Rayens W. Partial least squares for discrimination. J Chemo. 2003;17:166–173. 53. Sylvie Chevallier S, Bertrand D, Kohler A, Courcoux P. Application of PLS-DA in multivariate image analysis. J Chemo. 2006;20: 221–229. (TIF)</p

Results of principal component analysis using spectrogram data from all BKS.Cg db/db serum samples.

(A) PCA PC-1/PC-5 score plot diagram. Principal component 1 score was interpreted to be positively associated with week-old progression, whereas principal component 5 score was interpreted to be negatively and moderately associated with week-old progression. Furthermore, on closer examination of principal component 1, it shows a tendency to group with different score values for 10- and 14-week-old and 22- and 26-week-old mice. The 18-week-old mice are not uniform in score values but dispersed, with scores approximating the two groups. Furthermore, although the blood biochemistry test results shown in Table 2 varied from individual to individual, there is no clear correlation between this variation and the variation in the PCA score plots in Fig 3. (B) Principal component 1 loading diagram. The relative loading data are mapped onto the spectrogram area. The darker red and blue areas indicate the frequency components that are significantly related to the positive and negative contribution of each principal component. The threshold values, which were set in an exploratory manner based on the characteristics of the analyzed data to increase discriminability, were set above 0.4 and below −0.4. Values closer to 1 are shown in darker red, and values closer to −1 are shown in darker blue.</p

Graphical flow chart of matrixing of spectrogram data.

The process surrounded by the green box in S1 Fig is described in detail. For multivariate analysis of spectrogram data, a 256 × 1024 spectrogram (a) with frequency on the horizontal axis and time on the vertical axis was divided into 1024 single rows (b). All rows were re-arranged into a single row, and 1 × 262,144 single rows were reshaped (c). Since a single row represents spectrogram data for one individual (d), the single rows of spectrogram data for all individuals needed for multivariate analysis are combined to create a data matrix (e). (TIF)</p

H&E staining of carotid artery samples.

(A) Example of H&E staining of a carotid artery specimen from a 10-week-old BKS.Cg db/db mouse (B10-1), showing normal histology, with the tunica media composed of numerous smooth muscle cells. (B) Example of H&E staining of a carotid artery specimen from a 14-week-old BKS.Cg db/db mouse (B14-4), showing progressive intimal thickening compared with (A). (C) H&E staining of a carotid artery specimen from an 18-week-old BKS.Cg db/db mouse (B18-5), showing a more advanced disease stage and atheromatous plaques. (D) H&E staining of a carotid artery specimen from an 18-week-old Jcl:ICR mouse (J18-1) showing intimal thickening.</p

Results of partial least squares discriminant analysis performed on spectrogram data for serum samples from mice at 18 weeks of age only.

(A) Factor-1/Factor-2 score plot. Samples bisected by PCA in Fig 5 and those on the 10- and 14-week side are labeled the 18W A group and those on the 22- or 26-week side are labeled the 18W B group. Samples are clearly dichotomized by Factor-1 at 18W A and 18W B. (B) Factor-1 loading diagram.</p

Results of partial least squares discriminant analysis performed on spectrogram data for all serum samples of BKS.Cg db/db mice.

(A) Factor-1/Factor-2 score plot. Samples that were negatively distributed after PC-1 in Fig 3 are merged into the precursor group, and samples that were positively distributed are merged into the progressed group. Samples are clearly dichotomized by Factor-1. (B) Factor-1 loading diagram. The distribution is similar to the sum of Figs 6B and 7B.</p