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Additional file 1: Fig. S1. Differences in phylogenetic abundance between advanced adenoma patients (red) and control subjects (blue). The phylotypes that were increased (a, c) or decreased (b, d) in the advanced adenoma patients at the genus and species levels. Wilcoxon rank-sum tests were used to identify the differentially abundant genera and species. Fig. S2. Linear discriminant analysis (LDA) effect size (LEfSe) analysis revealed the genus difference (a) and species difference (b) in fecal microbiota between the advanced adenoma patients (negative score) and the control subjects (positive score). Fig. S3. The Bray-Curtis tree of 49 controls and 48 advanced adenoma patients and 300 disease cases from the previous papers. It was based on the top 30 genus abundance profiling. The 32 CRC samples were all in Hongkong and had the same branch as the 49 controls and the 48 advanced adenoma patients in the Bray-Curtis tree. Fig. S4. The relative abundance of the progressive gene markers in the five cohorts. Five cohorts were arranged on the horizontal axis, including discovery cohort 1, discovery cohort 2, validation cohort 1, validation cohort 2, and validation cohort 3. Each cohort has three groups (control group, adenoma group, and CRC group). All 277 progressive disease gene markers were arranged on the vertical axis. Fig. S5. The 15 progressive gene markers with decreased abundance (a) and 15 markers with increased abundance (b) were shown in the validation cohort. Some of the median values of the gene abundance were zero, however, the third quartile values of the gene abundance can reflect the expected trend. Fig. S6. The increased and decreased progressive fecal microbiota gene markers distributed in the KEGG pathways. Fig. S7. The progressive gene markers in the sulfur metabolism pathway. The genes with a red frame were up-regulated from the control group to the advanced adenoma group and then to the CRC group, and the genes with the green frame were down-regulated. Fig. S8. The progressive gene markers in taurine and hypotaurine metabolism pathway. The gene with a red frame was up-regulated from the control group to the advanced adenoma group and then to the CRC group. Fig. S9. The progressive gene markers in cysteine and methionine metabolism pathway. The genes with a red frame were up-regulated from the control group to the advanced adenoma group and then to the CRC group, and the genes with the green frame were down-regulated. Fig. S10. ROC curves of various effect indexes in discriminating patients with advanced adenoma and CRC from control subjects in the training cohort, respectively. (a) The ROC of Adenoma Effect Index 1 (AEI1) for advanced adenoma. (b) The ROC of Adenoma Effect Index 2 (AEI2) for adenoma with high-grade dysplasia. (c) The ROC of CRC Effect Index 1 (CEI1) for all stages of CRC. (d) The ROC of CRC Effect Index 2 (CEI2) for stage III-IV CRC. Fig. S11. Schematic diagram of the functions of the progressive microbiota gene markers. The increase of the H2S, decrease in cysteine, and accumulation of homocysteine of the gene markers are related to the occurrence and development of CRC. The biofilm regulator gene was also upregulated in colorectal adenoma and carcinoma, consistent with its roles in CRC carcinogenesis such as destroying the intestinal barrier and dysbiosis.

Shubin Wang,1,* Juan An,2,* Weiwei Dong,3,* Xin Wang,4 Jianqiu Sheng,4 Yan Jia,4 Yuqi He,4 Xianzong Ma,4 Jiheng Wang,4 Dedong Yu,1 Xiuqin Jia,1 Bingyu Wang,5 Wenbo Yu,5 Kejia Liu,5 Yuanyuan Zhao,6 Yun Wu,1 Wei Zhu,1 Yuanming Pan1,4,6 1Department of Oncology, Baotou City Central Hospital, Baotou 014040, People’s Republic of China; 2Department of Basic Research Medical Sciences, Qinghai University, Xining 810001, People’s Republic of China; 3Department of Oncology, General Hospital of Chinese People’s Liberation Army, Beijing 100085, People’s Republic of China; 4Department of Gastroenterology, The 7th Medical Center of Chinese PLA General Hospital, Beijing 100700, People’s Republic of China; 5Yidu Cloud (Beijing) Technology Co. Ltd 8F, Health Work, Beijing 100083, People’s Republic of China; 6National Center for Nanoscience and Technology, Zhongguancun, Beijing 100190, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yuanming PanNational Center for Nanoscience and Technology, Zhongguancun, Beijing 100190, People’s Republic of ChinaTel +86-10-6406-9831Fax +86-10-6401-2266Email panym@nanoctr.cnWei ZhuDepartment of Oncology, Baotou City Central Hospital, Baotou, Inner Mongolia 01404, People’s Republic of ChinaTel +86-472-6955473Fax +86-472-6955708Email 18686111667@163.comIntroduction: Glioma is the primary malignant brain tumor with poor prognosis. Berberine (BBR) was the potential drug for anti-tumor in glioma cells. Based on its limitation of poor aqueous solubility and instability, little information of BBR nanoparticles is reported in glioma.Methods: Different solutions including 5% glucose, 1*PBS, ddH2O, 0.9% NaCl, cell culture medium were selected, and only 5% glucose and ddH2O exhibited BBR-related nanoparticles. After heating for a longer time or adding a higher concentration of glucose solution, BBR nanoparticles were detected by TEM analysis. The uptake of BBR-Glu or BBR-Water nanoparticles were detected by immunofluorescence analysis for BBR autofluorescence. Cell viability was measured by MTT assay and Western blotting analysis. Apoptosis was performed with flow cytometric analysis and was detected by cleaved caspase-3 immuno-fluorescent staining. Cell cycle was used by flow cytometric analysis. Cytoskeleton was observed by confocal analysis using the neuron specific Class III ß-tubulin and ß-tubulin antibodies. Mitochondrial-related proteins were detected by Western blotting analyses and mito-tracker staining in live cells. Mitochondrion structures were observed by TEM analysis. ROS generation and ATP production were detected by related commercial kits. The tracking of BBR-Glu or BBR-Water nanoparticles into blood–brain barrier was observed in primary tumor-bearing models. The fluorescence of BBR was detected by confocal analyses in brains and gliomas.Results: BBR-Glu nanoparticles became more homogenized and smaller with dose- and time-dependent manners. BBR-Glu nanoparticles were easily absorbed in glioma cells. The IC50 of BBR-Glu in U87 and U251 was far lower than that of BBR-Water. BBR-Glu performed better cytotoxicity, with higher G2/M phase arrest, decreased cell viability by targeting mitochondrion. In primary U87 glioma-bearing mice, BBR-Glu exhibited better imaging in brains and gliomas, indicating that more BBR moved across the blood–brain tumor barrier.Discussion: BBR-Glu nanoparticles have better solubility and stability, providing a promising strategy in glioma precision treatment.Keywords: berberine, glioma, glucose-nanocarrier, nanoparticles, mitochondria

Supplemental Material for Career Development and Occupational Disease in Chinese Nurses: A Cross-Sectional Study by Wenyu Zhang, Xianzong Ma, Qian Xiao, Ran Miao, Mingna Zhang, and Xin Wang in INQUIRY: The Journal of Health Care Organization, Provision, and Financing.