Metastatic dissemination is the major cause of death from breast-cancer (BC). Fusobacterium nucleatum (F.n) is widely enriched in BC and has recently been identified as one of the high-risk factors for promoting BC metastasis. Here, with an experimental model, we demonstrated that intratumoral F.n induced BC aggressiveness by transcriptionally activating Epithelial-mesenchymal transition-associated genes. Therefore, the F.n may be a potential target to prevent metastasis. Given the fact that cancer-associated fibroblasts (CAFs) are abundant in BC and located near blood vessels, we report an optogenetic system that drives CAF to in situ produce human antibacterial peptide LL37, with the characteristics of biosafety and freely intercellular trafficking, for depleting intratumoral F.n, leading to a 72.1% reduction in lung metastatic nodules number without affecting the balance of the systemic flora. Notably, mild photothermal treatment was found that could normalize CAF, contributing to synergistically inhibiting BC metastasis. In addition, the system can also simultaneously encode a gene of TNF-related apoptosis-inducing ligand to suppress the primary tumor. Together, our study highlights the potential of local elimination of tumor pathogenic bacteria to prevent BC metastasis.

Objective:To observe the effect of interleukin-8 (IL-8) on the adhesion and migration of retinal vascular endothelial cells (RCEC).Methods:A cell experiment. Human RCEC (hRCEC) was divided into normal control group (N group), advanced glycation end product (AGE) treatment group (AGE group), and AGE-induced combined IL-8 antagonist SB225002 treatment group (AGE+SB group). The effect of AGE on IL-8 expression in hRCEC was observed by Western blot. The effect of SB225002 on hRCEC migration was observed by cell scratch assay. The effects of SB225002 on leukocyte adhesion and reactive oxygen species (ROS) on hRCEC were detected by flow cytometry. Student- t test was performed between the two groups. Oneway analysis of variance was performed among the three groups. Results:Compared with group N, the expression level of IL-8 in cells of AGE group was significantly increased, with statistical significance ( t=25.661, P<0.001). Compared with N group and AGE+SB group, cell mobility in AGE group was significantly increased ( F=29.776), leukocyte adhesion number was significantly increased ( F=38.159, 38.556), ROS expression level was significantly increased ( F=22.336), and the differences were statistically significant ( P<0.05). Conclusion:IL-8 antagonist SB225002 may down-regulate hRCEC adhesion and migration by inhibiting ROS expression.

ObjectiveTo study the conceptual framework and methodological system of the International Classification of Functioning, Disability and Health (ICF) in occupational therapy and its systematic implementation in clinical practice. MethodsBased on the ICF theory and the policy documents of the World Federation of Occupational Therapists, the conceptual framework of occupational therapy and the systematic implementation in clinical settings based on the ICF framework were analyzed. ResultsThis study constructed a conceptual framework and approach for occupational therapy based on ICF, and clarified the goals, principles, and implementation methods of integrated occupational therapy interventions in rehabilitation services. The goal of occupational therapy interventions was to improve the individual activity and participation through multidisciplinary and cross-cutting implementation of integrated occupational therapy programs to optimize functioning. Occupational therapy was based on the bio-psycho-social model, adhered to the principles of person-centeredness and functioning orientation, and implemented individualized intervention programs in different context. In clinical practice, it was recommended to follow ICF-based standardized process and systematically use World Health Organization Family International Classifications: functioning and unmet needs analysis using ICHI; functional classification, assessment and coding using ICF; disease classification, diagnosis and coding using ICD; intervention of occupational therapies using ICHI to build a systematic occupational therapy service system. ConclusionAn ICF-based occupational therapy concept and methodological system has been built, a comprehensive clinical occupational therapy implementation model has been established, the goal of activity and participation oriented occupational therapy interventions has been clarified, and the systematic, structured, standardized and refined level of occupational therapy has been enhanced.

Objective:To screening differentially expressed genes (DEGs) in proliferative diabetic retinopathy (DR) patients to provide new biological therapeutic targets for proliferative DR (PDR) therapy.Methods:A basic research. A total of 3 PDR patients (group PDR) and 3 non-diabetic patients (control group) were enrolled in the study in Tianjin Medical University Eye Hospital in October 2020. In addition, 40 cases of PDR and non-diabetic patients were selected and divided into PDR validation group and control validation group. Peripheral blood validation test was performed in PDR validation group and control validation group; RNA sequencing was performed in PDR group and control group. Transcriptomics (RNAseq) sequencing technology was used to screen DEG in PDR group and control group. The selected DEGs were analyzed by gene ontology (GO) function enrichment analysis, signal pathway enrichment analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) and protein-protein interaction network (PPI). The gene expression database was used to find the high-throughput data related to PDR, and multi queue comparison analysis was carried out. The target genes of differentially expressed miRNAs were predicted through targetscan platform, so as to clearly screen the correlation between DEG and PDR. Reverse transcription polymerase chain reaction and Western blot were used to verify the expression of DEG mRNA and protein related to PDR. The relative expression of PDR related DEG mRNA and protein between PDR validation group and control validation group were compared by paired t-test. Results:A total of 1 337 DEGs were screened by RNAseq sequencing in the peripheral blood of patients with PDR, of which 419 genes were up-regulated and 918 down-regulated. Among them, direct inhibitor of apoptosis protein-binding protein with low isoelectric point ( DIABLO), zinc finger and BTB domain containing 10 ( ZBTB10), polo-like kinases 3 ( PLK3), regulatory subunit 1 ( PIK3R1) and B cell translocation gene 3 (BTG3) were differentially expressed in PDR patients. The function of GO was enriched from the analysis of molecular function, biological process and cellular composition. The results showed that DIABLO, ZBTB10, PLK3, PIK3R1, BTG3 were involved in the pathological process related to PDR. KEGG enrichment analysis showed that glucose metabolic pathways such as extracellular matrix receptors, cytokine regulatory pathway, p53 signal pathway and galactose metabolism may be involved in the process of differential genes. The analysis of PPI protein interaction network showed that the larger the DEG-associated protein node, the greater the number of associated nodes. Among them, DIABLO, ZBTB10, PLK3, PIK3R1 and BTG3 played significant roles in the formation of the action network. By comparing and analyzing the existing high-throughput data related to diabetic retinopathy in Gene Expression Omnibus database and predicting by Targetscan platform, it was found that some significant differences in miRNA reported in aqueous humor, vitreous fluid and plasma of DR patients can be regulated by the differential genes found in this study. Compared with the control verification group, the relative expressions of DIABLO, ZBTB10, PLK3, PIK3R1 mRNA and protein in peripheral blood of the PDR verification group were up-regulated, and the relative expression of BTG3 mRNA and protein was down-regulated. Conclusion:DIABLO, ZBTB10, PLK3, PIK3R1 and BTG3 are DEGs in patients with PDR, and they can participate in the disease process by regulating the biological processes of cell proliferation, fibrosis and oxidative stress.

Objective:To observe the effect of bone morphogenetic protein 4 (BMP4) on the proliferation and migration of human retinal microvascular endothelial cells (hRMEC) under oxidative stress.Methods:The hRMEC cultured in vitro were divided into control group, 4-hydroxynonenal (HNE) treatment group (4-HNE group), 4-HNE+BMP4 group (BMP4 group). Cell culture medium of 4-HNE treatment group was added with 10 μmmol/L 4-HNE; cell culture of BMP4 group was cultured with 10 μmmol/L 4-HNE, and after stimulation for 6 h, 100 ng/ml recombinant human BMP4 was added. The effects of 4-HNE and BMP4 on hRMEC viability was detected by thiazole blue colorimetric method. The effects of 4-HNE and BMP4 on cell migration was determined by cell scratch test. The relative expression of BMP4 mRNA in the cells of the control group and 4-HNE treatment group and the mRNA expression of the control group, the fibronectin (FN) of BMP4 group, laminin (Laminin), α-smooth muscle contractile protein (α-SMA), and collagen type Ⅰ (Collagen Ⅰ), vascular endothelial growth factor (VEGF), and connective tissue growth factor (CTGF) were detected by real-time quantitative polymerase chain reaction (qRT-PCR). Western blot was used to detect the relative expression of BMP4 protein in the control group and 4-HNE group. The control group and 4-HNE group were compared by t test. Results:Compared with the control group, cell viability ( t=12.73, 16.26, P=0.000 2, <0.000 1), cell migration rate ( t=28.17, 37.48, P<0.000 1, <0.000 1) in 4-HNE group and BMP4 group were significantly increased, and the difference was statistically significant; the relative expression of BMP4 mRNA and protein in the 4-HNE group was significantly increased, and the difference was statistically significant ( t=16.36, 69.35, P=0.000 1, <0.000 1). The qRT-PCR test results showed that compared with the control group, the relative expression of VEGF, FN, Laminin, α-SMA, Collagen Ⅰ, and CTGF mRNA in the cells of the BMP4 group was significantly increased, and the difference was statistically significant ( t=10.61, 17.00, 14.85, 7.78, 12.02, 10.61, P=0.0004, <0.000 1, 0.000 1, 0.001 5, 0.000 1, 0.000 4). Conclusion:BMP4 can induce the proliferation and migration of hRMEC; it can also regulate the expression of angiogenesis factors and fibrosis-related factors in hRMEC.

Objective:To investigate the effects of interferon gene stimulating protein (STING) inhibitor (C176) on human retinal microvascular endothelial cells (hRMEC) under oxidative stress.Methods:An animal experimental study. In vivo experiment: 48 healthy male C57BL/6J mice were randomly divided into wild type mice group (WT group) and diabetes (DM) group, with 24 mice in each group. DM mice were induced by streptozotocin to establish DM model. After successful modeling, DM group was divided into DM+dimethyl sulfoxide (DMSO) group and DM+C176 group, with 12 mice in each group. The mice in the DM+DMSO group were intraperitoneally injected with DMSO at the dose of 50 mg/kg. Mice in DM+C176 group were intraperitoneally injected with STING inhibitor C176 750 nmol at the dose of 50 mg/kg. Four weeks after modeling, immunohistochemical staining, Western blot and real-time fluorescence quantitative polymerase chain reaction were used to detect the expression of STING in the retina of WT and DM mice. The leukocyte adhesion test was used to detect the number of leukocytes adhering to hRMEC in mice with WT, DM+DMSO and DM+C176 groups. In vitro experiment: hRMEC was randomly divided into conventional culture cell group (N group), dimethyl sulfoxide (DMSO) group (with DMSO intervention) and C176 group (with C176 intervention). The cells were induced by 150 μg/ml glycation end products for each group. In vitro leukocyte adhesion test combined with 4', 6-diamino-2-phenylindole staining was used to detect the number of leukocytes adhering to hRMEC. The adherent leukocytes were quantitatively analyzed by flow cytometry; H 2DCFDA/reactive oxygen species (ROS) fluorescence probe was used to detect ROS expression in cells; Seahorse XFe96 cell energy metabolism analyzer was used to measure the level of intracellular glycolysis. t-test was used to compare the two groups; single factor analysis of variance was used to compare the three groups. Results:In vivo experiment: compared with WT group, the expression level of STING ( t=73.248) and the relative expression amount of mRNA ( t=67.385) in the retina of DM group mice increased significantly ( P<0.05). Compared with WT group, the number of leukocytes adhering to the retinal vessels of mice in DM+DMSO group was significantly increased, while that in DM+C176 group was significantly decreased ( F=84.352, P<0.01). In vitro: compared with N group and DMSO group, the number of leukocyte adhesion on hRMEC in C176 group decreased significantly ( F=35.251, P<0.01). Compared with N group, the number of leukocytes adhering to hRMEC in DMSO group and C176 group decreased significantly ( F=26.374, P<0.01). The ROS level in hRMEC in C176 group was significantly lower than that in N group and C176 group ( F=41.362, P<0.01). Compared with N group and DMSO group, the glycolysis level of hRMEC in C176 group was significantly reduced, with a statistically significant difference ( F=68.741, P<0.01). Conclusion:Inhibiting the expression of STING in retinal vascular endothelial cells can improve the progress of DM by inhibiting leukocyte adhesion, ROS production and glycolysis level.

Objective To evaluate the role of Toll-like receptor 4 ( TLR4)/nuclear factor kappa B ( NF-κB) signaling pathway in the development of postoperative chronic pain and the relationship with ex-pression of voltage-gated sodium channel 1. 7 (Nav 1. 7) in the dorsal root ganglion (DRG) of rats. Meth-ods Thirty-six clean-grade healthy male Sprague-Dawley rats, aged 9-11 weeks, weighing 200-250 g, were divided into 3 groups ( n=12 each) using a random number table method: control group ( group C) , normal saline group ( group NS) and TLR4 antagonist LPS-RS group ( group R) . Postoperative chronic pain was produced by skin/muscle incision and retraction (SMIR). From 1 day before SMIR to 10 days after SMIR, NS group received continuous intrathecal injection of normal saline 10μl, while R group received continuous intrathecal injection of LPS-RS 20μg/10μl. Six rats were randomly selected in each group, and the mechanical pain threshold was measured at 1 day before SMIR and 1, 5, 10, 15 and 20 days after SMIR. The 6 rats left in each group were sacrificed at day 10 after SMIR, and the DRGs of the lumbar seg-ment (L4,5) were removed for determination of the expression of phosphorylated NF-κB (p-NF-κB) and Nav 1. 7. Results Compared with group C, the mechanical pain threshold was significantly decreased at 5-20 days after SMIR, and the expression of p-NF-κB and Nav1. 7 was up-regulated at 10 days after SMIR in group NS ( P＜0. 01) . Compared with group NA, the mechanical pain threshold was significantly increased at 5-20 days after SMIR, and the expression of p-NF-κB and Nav 1. 7 was down-regulated at 10 days after SMIR in group R (P＜0. 01). Conclusion Up-regulated expression of Nav1. 7 in DRGs after activating TLR4/NF-κB signaling pathway is involved in the development of postoperative chronic pain in rats.

OBJECTIVE:To provide basic industrial research and development recommendations for the supply-side structural reform in drug distribution in China. METHODS:Structure-conduct-performance(SCP)analysis paradigm from Harvard was used for in-depth study of market structure,business conduct and business performance in China since 2010. RESULTS:Since 2010, drug distribution industry in China had shown the scale of decline in the growth rate,low market concentration,increased barriers to entry,the leading of state-owned capital,etc. Though corporate mergers and acquisitions and public financing expanded the scale,management information,third-party logistics,Internet+and pharmacy alliance business model flourished,the industry op-erating performance was poor,state-owned enterprises showed poor performance. CONCLUSIONS:The transformation and upgrad-ing of the industry business model should be promoted by targetedly accelerating the supply-side structural reforms in the drug distri-bution industry,encouraging corporate finance mergers,breaking local protectionism,developing cooperative pharmacies and phar-maceutical third-party logistics alliances and leveraging the Internet.

Huolinguole lignite was sequentially extracted with carbon disulfide, ethyl acetate, methanol and acetone.All of the extracts were analyzed using a time-of-flight mass spectrometry (TOF-MS) equipped with an atmospheric pressure photoionization (APPI) ion source.Toluene or 1,4-difluorobenzene was chosen as dopant for APPI.The results indicated that both dopants could well ionize compounds which could not be ionized by APPI without dopant.Toluene induced higher ionization efficiency than 1,4-difluorobenzene.Some compounds in the extracts were identified as dimers, which might be formed via molecular association.Heteroatoms were identified in all of the associated molecules.Molecular weight distributions under three APPI ionization modes were similar.Compounds with molecular weight from 200 to 500 Da occupied 60% of all the products and around 10% of the products had molecular weight over 500 Da.

OBJECTIVE:To provide reference for evaluating the new round of essential medicine bidding and purchase policy in China,ensuring the stability of essential medicine supply and improving national essential medicine system.METHODS:Based on the theoretical analysis,taking Shanghai and Yunan province as sample,the questionnaire was designed to investigate the difficulty to implement essential drug purchase policy in essential medicine supply system,and the bidding situation evaluation of production enterprises by both sides of local drug bidding among staff in production enterprises,circulation enterprises,primary health care institutions and other institutions.The types of influential factors for the implementation of essential medicine purchase policy were determined finally.Factor analysis method was used to screen the key influential factors.RESULTS & CONCLUSIONS:Four common factors had a great influence on the implementation of essential medicine policy in China.The impact of the large to small is the rationality of the tender procurement program,the standardization of pharmaceutical production,the stability of demand for drug use and macroeconomic policy regulation and protection.It is suggested to further improve the drug recruitment system,select the products that have quality assurance and production specifications,and fully meet the diverse needs of the market,while provide relatively stable usage information for manufacturers.If necessary,"drug purchase with target quantity" of Shanghai model can be forced to carry out by policy so as to ensure that the interests of all parties in the pharmaceutical supply chain are treated fairly.