Manipulating the Subcellular Localization and Anticancer Effects of Benzophenothiaziniums by Minor Alterations of N-Alkylation.

Cationic, water-soluble benzophenothiaziniums have been recognized as effective type I photosensitizers (PSs) against hypoxic tumor cells. However, the study of the structure-property relationship of this type of PS is still worth further exploration to achieve optimized photodynamic effects and minimize the potential side effects. Herein, we synthesized a series of benzophenothiazine derivatives with minor N-alkyl alteration to study the effects on the structure-property relationships. The cellular uptake, subcellular organelle localization, reactive oxygen species (ROS) generation, and photocytotoxicity performances were systematically investigated. NH2NBS and EtNBS specifically localized in lysosomes and exhibited high toxicity under light with a moderate phototoxicity index (PI) due to the undesirable dark toxicity. However, NMe2NBS with two methyl substitutions accumulated more in mitochondria and displayed an excellent PI value with moderate light toxicity and negligible dark toxicity. Without light irradiation, NH2NBS and EtNBS could induce lysosomal membrane permeabilization (LMP), while NMe2NBS showed no obvious damage to lysosomes. After irradiation, NH2NBS and EtNBS were released from lysosomes and relocated into mitochondria. All compounds could induce mitochondria membrane potential (MMP) loss and nicotinamide adenine dinucleotide phosphate (NADPH) consumption under light to cause cell death. NMe2NBS exhibited remarkable in vivo photodynamic therapy (PDT) efficacy in a xenograft mouse tumor (inhibition rate, 89%) with no obvious side effects. This work provides a valuable methodology to investigate the structure-property relationships of benzophenothiazine dyes, which is of great importance in the practical application of PDT against hypoxia tumor cells.

An Endoplasmic Reticulum Targeting Type I Photosensitizer for Effective Photodynamic Therapy against Hypoxic Tumor Cells.

Organelle-targeted type I photodynamic therapy (PDT) shows great potential to overcome the hypoxic microenvironment in solid tumors. The endoplasmic reticulum (ER) is an indispensable organelle in cells with important biological functions. When the ER is damaged due to the production of reactive oxygen species (ROS), the accumulation of misfolded proteins will interfere with ER homeostasis, resulting in ER stress. Here, an ER-targeted benzophenothiazine-based photosensitizer NBS-ER was presented. ER targeting modification significantly reduced the dark toxicity and improved phototoxicity index (PI). NBS-ER could effectively produce O2 - ⋅ with near-infrared irradiation, making its phototoxicity under hypoxia close to that under normoxia. Meanwhile, the photoinduced ROS triggered ER stress and induced apoptosis. In addition, NBS-ER possessed excellent photodynamic therapeutic effect in 4T1-tumor-bearing mice.

Ferroptosis Photoinduced by New Cyclometalated Iridium(III) Complexes and Its Synergism with Apoptosis in Tumor Cell Inhibition.

Limited therapeutic efficacy to hypoxic and refractory solid tumors has hindered the practical application of photodynamic therapy (PDT). Two new benzothiophenylisoquinoline (btiq)-derived cyclometalated IrIII complexes, IrL1 and MitoIrL2, were constructed as potent photosensitizers, with the latter being designed for mitochondria accumulation. Both complexes demonstrated a type I PDT process and caused photoinduced ferroptosis in tumor cells under hypoxia. This ferroptosis featured lipid peroxide accumulation, mitochondria shrinkage, down-regulation of glutathione peroxidase 4 (GPX4), and ferrostatin-1 (Fer-1)-inhibited cell death. Upon photoirradiation under hypoxia, mitochondria targeting MitoIrL2 caused mitochondria membrane potential (MMP) collapse, ATP production suppression, and induced cell apoptosis. The synergetic effect of ferroptosis and apoptosis causes MitoIrL2 to outperform IrL1 in inhibiting the growth of MCF-7, PANC-1, MDA-MB-231 cells and multicellular spheroids. This study demonstrates the first example of ferroptosis induced by photosensitizing IrIII complexes. Moreover, the synergism of ferroptosis and apoptosis provides a promising approach for combating hypoxic solid tumors through type I PDT processes.

Flexible π-Conjugated 2,5-Diarylamino-Terephthalates: A New Class of Mechanochromic Luminophores with Tunable Aggregation States.

The generation of different thermodynamically (meta)stable states is crucial for the development of smart solid-state luminescent materials. However, the design of luminophores with tunable aggregation states is remaining a grand challenge. Herein, we present a family of mechanochromic luminophores with tunable metastable states, based on the dynamically controllable π-π stacking of the flexible π-conjugated structure of 2,5-diarylamino-terephthalates in the solid state. The experimental data revealed that both the kinetically controlled metastable state and thermodynamic controlled stable state can be generated via tuning the intermolecular interactions such as π-π stacking and hydrogen bonds. As a result, the highly sensitive mechano-stimuli response of these luminophores was successfully achieved.

Reversible FRET Fluorescent Probe for Ratiometric Tracking of Endogenous Fe3+ in Ferroptosis.

A reversible fluorescent probe DRhFe is devised by linking spirolactam rhodamine and dansylamide through an Fe3+ ionophore N2-hydroxyethyldiethylenetriamine, where Fe3+ chelation generating rhodamine B can switch intramolecular fluorescence resonance energy transfer for ratiometric Fe3+ sensing. Probe DRhFe exhibits an Fe3+-specific emission shift from 483 to 576 nm, and this ratiometric response has been successfully applied to monitor labile Fe3+ fluctuations in cells undergoing ferroptosis.

[Clinical effect of Saccharomyces boulardii powder combined with azithromycin sequential therapy in treatment of children with diarrhea secondary to Mycoplasma pneumoniae pneumonia].

OBJECTIVE: To investigate the clinical effect of Saccharomyces boulardii powder combined with azithromycin sequential therapy in the treatment of children with diarrhea secondary to Mycoplasma pneumoniae pneumonia. METHODS: A total of 88 children with diarrhea secondary to Mycoplasma pneumoniae pneumonia between June 2015 and March 2017 were divided into control group and study group using a random number table, with 44 children in each group. The children in the control group were given routine treatment combined with azithromycin sequential therapy, and those in the study group were given oral Saccharomyces boulardii powder in addition to the treatment in the control group until the end of azithromycin sequential therapy. After the treatment ended, the two groups were compared in terms of time to improvement of clinical symptoms, length of hospital stay, clinical outcome, defecation frequency before and after treatment, condition of intestinal dysbacteriosis, and incidence of adverse events. RESULTS: Compared with the control group, the study group had significantly shorter time to improvement of clinical symptoms and length of hospital stay (P<0.05). The study group had a significantly higher response rate than the control group (P<0.05). On days 3 and 5 of treatment, the study group had a significant reduction in defecation frequency compared with the control group (P<0.05). The study group had a significantly lower rate of intestinal dysbacteriosis than the control group (P<0.05). There was no significant difference in the incidence of adverse events between the two groups (P>0.05). CONCLUSIONS: In the treatment of children with diarrhea secondary to Mycoplasma pneumoniae pneumonia, Saccharomyces boulardii powder combined with azithromycin sequential therapy can improve clinical symptoms, shorten the length of hospital stay, reduce defecation frequency and the incidence of intestinal dysbacteriosis, and improve clinical outcomes, and does not increase the risk of adverse events.

Efficacy and prognostic analysis of carbon nanotracers combined with the da Vinci robot in the treatment of esophageal cancer.

BACKGROUND: Traditional methods cannot clearly visualize esophageal cancer (EC) tumor contours and metastases, which limits the clinical application of da Vinci robot-assisted surgery. AIM: To investigate the efficacy of the da Vinci robot in combination with nanocarbon lymph node tracers in radical surgery of EC. METHODS: In total, 104 patients with early-stage EC who were admitted to Liuzhou worker's Hospital from January 2020 to June 2023 were enrolled. The patients were assigned to an observation group (n = 52), which underwent da Vinci robot-assisted minimally invasive esophagectomy (RAMIE) with the intraoperative use of nanocarbon tracers, and a control group (n = 52), which underwent traditional surgery treatment. The operation time, intraoperative blood loss, postoperative drainage tube indwelling time, hospital stay, number of lymph nodes dissected, incidence of complications, and long-term curative effects were comparatively analyzed. The postoperative stress response C-reactive protein (CRP), cortisol, epinephrine (E) and inflammatory response interleukin (IL)-6, IL-8, IL-10, and tumor necrosis factor-alpha (TNF-α) were evaluated. RESULTS: Compared with the control group, the observation group had significantly lower postoperative CRP, cortisol, and E levels (P < 0.05) with a milder inflammatory response, as indicated by lower IL-6, IL-10, and TNF-α levels (P < 0.05). Patients who underwent RAMIE had less intraoperative blood loss and shorter operation times and hospital stays than those who underwent traditional surgery. The average number of dissected lymph nodes, time of lymph node dissection, and mean smallest lymph node diameter were all significantly lower in the observation group (P < 0.05). The rate of postoperative complications was 5.77% in the observation group, significantly lower than the 15.38% observed in the control group. Furthermore, the lymphatic metastasis rate, reoperation rate, and 12- and 24-month cumulative mortality in the observation group were 1.92%, 0%, 0%, and 0%, respectively, all of which were significantly lower than those in the control group (P < 0.05). CONCLUSION: The treatment of EC using the da Vinci robot combined with nanocarbon lymph node tracers can achieve good surgical outcomes and demonstrates promising clinical applications.

Composite polymer electrolyte based on poly(vinylidene fluoride-hexafluoropropylene) (PVDF-HFP) for solid-state batteries.

Using solid-state electrolytes (SSEs) with excellent thermal and electrical stability to replace liquid electrolytes, and assembling solid-state lithium-ion batteries (SSLIBs) is considered the best solution to these safety issues. However, it is difficult for a single electrolyte to have the characteristics of high ionic conductivity, low interface resistance, and high stability of the counter electrode at the same time. In this work, the composite polymer electrolyte membrane (CPE) of inorganic Li1.3Al0.3Ti1.7(PO4)3 (LATP) and organic poly(vinylidene fluoride-hexafluoropropylene) (PVDF-HFP) polymer was successfully prepared by traditional casting method. The addition of LATP (10 wt %) ceramic powder makes CPE membrane (CPE-10) exhibit excellent electrochemical performance: the lithium-ion transference number and electrochemical window are as high as 0.60 and 4.94 V, respectively. Moreover, the CPE-10 showed excellent Li-metal stability, thereby enabling the Li-Li symmetric cells to stably run for over 300 h at 0.1 mA/cm2 with effective lithium dendrite inhibition. When paired with a high-voltage LiNi0.6Co0.2Mn0.2O2 (NCM622) cathode, the Li/CPE-10/NCM622 cell exhibited excellent electrochemical performance: the highest specific discharge capacity of 152 mAh/g could be conducted at 0.2C after 50 cycles corresponding to 100% Coulombic efficiencies. The prepared CPE-10 demonstrates excellent electrochemical performance, providing an effective design strategy for SSLMBs.

[Medium and Long-term Carbon Emission Projections and Emission Reduction Potential Analysis of the Lingang Special Area Based on the LEAP Model].

Based on the existing statistical data of the Lingang Special Area in Shanghai and considering its future socio-economic development， industrial structure， and technological development， a LEAP-Lingang model was developed to analyze the evolution trends of energy demand and carbon emissions under the baseline scenario， low-carbon scenario， and enhanced low-carbon scenario. To enhance the prediction accuracy of the model， the Logistic population growth model was used to predict future population data， and the learning curve model was used to simulate the cost evolution trend of related carbon reduction technologies. In addition， an economic evaluation model for carbon reduction technologies was developed， and the economic costs and emission reduction potential of typical carbon reduction technologies were evaluated by drawing a marginal emission reduction cost curve. The results showed that under the enhanced low-carbon scenario， the renewable energy accounted for 69% of the primary energy consumption， and the electric energy accounted for 91% of the terminal energy demand in 2060. The Lingang Special Area could achieve carbon peak by 2030， and the carbon emissions in 2060 were predicted to decrease by 94% compared to that in the baseline scenario. In terms of contribution to emission reduction， clean energy substitution， industrial structure optimization， and terminal energy efficiency improvement played a key role in reducing carbon emissions near the port. In the medium term （until 2035）， they were predicted to contribute 35.1%， 27.3%， and 16.2% of carbon emissions， respectively， and in the long term （until 2060）， they should contribute 50.6%， 8.75%， and 7.7% of carbon emissions， respectively. Regarding specific carbon reduction technologies， hydrogen power generation； water electrolysis for hydrogen； and carbon capture， utilization， and storage （CCUS） technology were of great significance for achieving net-zero emissions， but the costs of emission reduction were relatively high. The research results can provide ideas and references for the low-carbon and green development of the Lingang Special Area and related areas.

Aggregation-Induced Emission Luminogens for Enhanced Photodynamic Therapy: From Organelle Targeting to Tumor Targeting.

Photodynamic therapy (PDT) has attracted much attention in the field of anticancer treatment. However, PDT has to face challenges, such as aggregation caused by quenching of reactive oxygen species (ROS), and short 1O2 lifetime, which lead to unsatisfactory therapeutic effect. Aggregation-induced emission luminogen (AIEgens)-based photosensitizers (PSs) showed enhanced ROS generation upon aggregation, which showed great potential for hypoxic tumor treatment with enhanced PDT effect. In this review, we summarized the design strategies and applications of AIEgen-based PSs with improved PDT efficacy since 2019. Firstly, we introduce the research background and some basic knowledge in the related field. Secondly, the recent approaches of AIEgen-based PSs for enhanced PDT are summarized in two categories: (1) organelle-targeting PSs that could cause direct damage to organelles to enhance PDT effects, and (2) PSs with tumor-targeting abilities to selectively suppress tumor growth and reduce side effects. Finally, current challenges and future opportunities are discussed. We hope this review can offer new insights and inspirations for the development of AIEgen-based PSs for better PDT effect.

The Interaction Between Autophagy and JAK/STAT3 Signaling Pathway in Tumors.

Tumor is one of the important factors affecting human life and health in today's world, and scientists have studied it extensively and deeply, among which autophagy and JAK/STAT3 signaling pathway are two important research directions. The JAK/STAT3 axis is a classical intracellular signaling pathway that assumes a key role in the regulation of cell proliferation, apoptosis, and vascular neogenesis, and its abnormal cell signaling and regulation are closely related to the occurrence and development of tumors. Therefore, the JAK/STAT3 pathway in tumor cells and various stromal cells in their microenvironment is often considered as an effective target for tumor therapy. Autophagy is a process that degrades cytoplasmic proteins and organelles through the lysosomal pathway. It is a fundamental metabolic mechanism for intracellular degradation. The mechanism of action of autophagy is complex and may play different roles at various stages of tumor development. Altered STAT3 expression has been found to be accompanied by the abnormal autophagy activity in many oncological studies, and the two may play a synergistic or antagonistic role in promoting or inhibiting the occurrence and development of tumors. This article reviews the recent advances in autophagy and its interaction with JAK/STAT3 signaling pathway in the pathogenesis, prevention, diagnosis, and treatment of tumors.

BODIPY-based monofunctional Pt (II) complexes for specific photocytotoxicity against cancer cells.

Photodynamic therapy (PDT) has attracted extensive attention in cancer treatment because of its minimum trauma, less side effects, and so on. Photosensitizers, as one of the core elements of PDT, usually have to face problems such as poor water solubility and light stability, lack of targeting, and other problems, which seriously affect the therapeutic effect. In this work, two BODIPY (boron-dipyrromethene)-based monofunctional Pt (II) complexes, 1a and 2a, were designed and synthesized, and their PDT effect was studied. The Pt atom improved the singlet oxygen quantum yield (0.19 for 1a and 0.14 for 2a, respectively), which effectively improves the efficiency of PDT. MTT assay confirmed that the short time photo-irradiation distinctly promoted antitumor cytotoxicity of Pt (II) compounds against different cell lines. For 1a under irradiation, the IC50 value of cancer cell lines were 13.1 µM for HeLa cells and 7.6 µM for MCF-7 cells, while those of normal cell lines were 32.4 µM for HBL-100 cells and 48.6 µM for L02 cells. The results demonstrated that 1a showed specific phototoxicity to cancer cells. This specific selectivity could be attributed to the synergistic effect of increased cellular uptake (determined by ICP-MS) and higher ROS generation (detected by Cell ROX Deep Red) in cancer cells after irradiation. This study laid the foundation for the future design and synthesis of effective PDT photosensitizers.

[Oral microbiological diversity in patients with salivary adenoid cystic carcinoma].

OBJECTIVE: The aim of this study was to identify the differences in microbial diversity and community in patients with salivary adenoid cystic carcinoma (SACC). METHODS: Saliva was collected from 13 patients with SACC confirmed by histopathological diagnosis and 10 healthy control subjects. Total metagenomic DNA was extracted. The DNA amplicons of the V3-V4 hypervariable regions of the 16S rRNA gene were generated and subjected to high-throughput sequencing. Microbial diversity and community structure were analyzed with Mothur software. RESULTS: A total of 16 genera of dominant bacteria in the SACC group were found, including Streptococcus (36.68%), Neisseria (8.55%), Prevotella_7 (7.53%), and Veillonella (6.37%), whereas 15 dominant bacteria in the control group were found, including Streptococcus (18.41%), Neisseria (18.20%), Prevotella_7 (8.89%), Porphyromonas (6.20%), Fusobacterium (5.86%) and Veillonella (5.82%). The statistically different phyla between the two groups were Firmicutes, Proteobacteria and Fusobacterium (P<0.05). The statistically different genera between the two groups were Streptococcus, Neisseria and Porphyromonas (P<0.05), and Capnocytophaga was only detected in patients with SACC. CONCLUSIONS: Significant differences were observed in the oral microorganisms between the two groups.

[Expression patterns of ectodysplasin and ectodysplasin receptor during early dental development in zebrafish].

OBJECTIVE: This study aims to study the expression patterns of ectodysplasin (EDA) and ectodysplasin receptor (EDAR) during the early development of zebrafish and provide a foundation for further research of the Eda signaling pathway in tooth development. METHODS: Total RNA was extracted from zebrafish embryos at 48 hours postfertilization (hpf) and then reverse transcribed for cDNA library generation. The corresponding RNA polymerase was selected for the synthesis of the digoxin-labeled antisense mRNA probe of zebrafish pharyngeal tooth specific marker dlx2b and Eda signaling-associated genes eda and edar in vitro. The three sequences were ligated into a pGEMT vector with a TA cloning kit, and polymerase chain reaction (PCR) was applied to linearize the plasmid. The resultant PCR sequences were used as templates for synthesizing Dig-labeled mRNA probe dlx2b, eda, and edar. Zebrafish embryos were collected at 36, 48, 56, 60, 72, and 84 hpf, then whole mount in situ hybridization was performed for the detection of eda and edar expression patterns. Then, their expression patterns at 72 hpf were compared with the expression pattern of dlx2b. RESULTS: The mRNA antisense probes of dlx2b, eda, and edar were successfully obtained. The positive signals of eda and edar were observed in zebrafish pharyngeal tooth region at 48-72 hpf and thus conform to the signals of dlx2b in the positive regions. CONCLUSIONS: The ligand eda and edar, which are associated with the Eda signaling pathway, are strongly expressed only at the pharyngeal tooth region in zebrafish from tooth initiation to the morphogenesis stage. Thus, the Eda signaling pathway may be involved in the regulation of the early development of zebrafish pharyngeal teeth.

Immunoregulatory role of IL-2/STAT5/CD4+CD25+Foxp3 Treg pathway in the pathogenesis of chronic osteomyelitis.

BACKGROUND: This study aimed to investigate immunoregulatory role of IL-2/STAT5/CD4+CD25+Foxp3 Treg pathway in pathogenesis of chronic osteomyelitis (COM). METHODS: Sprague-Dawley (SD) rats were injected with Staphylococcus aureus to establish COM model. 4 weeks later, the lesioned bones were collected and subjected to HE staining for examination of inflammatory infiltration. Enzyme-linked immunosorbent assay (ELISA) was employed to detect IL-2 expression in peripheral blood; flow cytometry was performed to detect CD25+CD4+Foxp3 Treg cells in peripheral blood. The mRNA expression of Foxp3 and CTLA-4 was detected by RT-PCR and the protein expression of STAT5 and p-STAT5 was detected by Western Blotting in CD25+CD4+Foxp3 Treg cells. RESULTS: In COM group, the periosteal thickening was observed in femur, and there were a large number of inflammatory cells in medullary cavity, accompanied by bone destruction. At 1, 2 and 4 weeks, IL-2 expression significantly increased, the proportion of CD4+CD25+FoxP3 Treg cells in peripheral monocytes markedly increased, the mRNA expression of Foxp3 and CTLA-4 and p-STAT5 protein expression increased dramatically in Treg cells as compared to control group (P<0.001). CONCLUSIONS: IL-2/STAT5/CD4+CD25+Foxp3 Treg pathway may be involved in the pathogenesis of COM, and excessive immunosuppression may lead to persistent infectious inflammation, which may become a key target for future treatment of COM.

[Generation of bone morphogenetic protein 4 conditional RNA interference mice].

To explore bone morphogenetic protein 4 (BMP4) function in the developing bone, a BMP4 conditional RNA interference (CRNAi) vector was constructed based on the pBSK/U6 vector with a LoxPneo cassette. The transgene fragment targeting bmp4 was obtained by Kpn and Afl double digestion and was purified before being microinjected into fertilized eggs from FVB/NJ mice. BMP4CRNAi transgenic mice were genotyped by PCR. And the PCR positive mice were crossed with Col2a1-Cre transgenic mice, whose Cre recombinase was specifically expressed in osteo-chondro-progenitor cells. Bmp4 mRNA expression in primary chondrocytes were examined by semi-quantitive RT-PCR to determine RNA interference efficiency. Results showed that BMP4(CRNAi) mice and BMP4 (Col2a1-CRNAi) mice were produced successfully, and bmp4 knockdown efficiency in primary chondrocytes of BMP4 Col2a1-CRNAi mice was 81%. This transgenic mouse line provides excellent model for studying the role of BMP4 in chondrocyte development, and BMP4CRNAi mouse may be a good model for studying the role of BMP4 in different cells, tissues and organs through crossing with different Cre transgenic mice.

Domino-like multi-emissions across red and near infrared from solid-state 2-/2,6-aryl substituted BODIPY dyes.

Considerable achievements on multiple emission capabilities and tunable wavelengths have been obtained in inorganic luminescent materials. However, the development of organic counterparts remains a grand challenge. Herein we report a series of 2-/2,6-aryl substituted boron-dipyrromethene dyes with wide-range and multi-fluorescence emissions across red and near infrared in their aggregation states. Experimental data of X-ray diffraction, UV-vis absorption, and room temperature fluorescence spectra have proved the multiple excitation and easy-adjustable emission features in aggregated boron-dipyrromethene dyes. Temperature-dependent and time-resolved fluorescence studies have indicated a successive energy transfer from high to step-wisely lower-located energy levels that correspond to different excitation states of aggregates. Consistent quantum chemical calculation results have proposed possible aggregation modes of boron-dipyrromethene dyes to further support the above-described scenario. Thus, this study greatly enriches the fundamental recognition of conventional boron-dipyrromethene dyes by illustrating the relationships between multiple emission behaviors and the aggregation states of boron-dipyrromethene molecules.

Optical properties and mechanofluorochromism of new BODIPY dyes based on the pyridine-pyrimidine hybrid structure.

A new family of BODIPY dyes (PPB1-4) based on the pyridine-pyrimidine hybrid structure has been synthesized. These dyes show high cyan fluorescence in hexane with quantum yields (Φf) ranging from 0.33 to 0.90. Interestingly, moderate fluorescence of PPB1-4 is observed in the solid state (Φf, 0.11-0.26). X-ray structure analysis demonstrates that H-aggregation should be responsible for the red-shifted and quenched emission in the solid state. Moreover, PPB1-4 show high contrast and reversible mechanochromic fluorescence based on the proposed mechanism of mechanical stimuli triggered dimer dissolution and formation. In addition, rewritable mechanochromic fluorescence recording media fabricated with PPB4 have been successfully achieved.

Rifampicin-induced injury in HepG2 cells is alleviated by TUDCA via increasing bile acid transporters expression and enhancing the Nrf2-mediated adaptive response.

Bile acid transporters and the nuclear factor erythroid 2-related factor (Nrf-2)-mediated adaptive response play important roles in the development of drug-induced liver injury (DILI). However, little is known about the contribution of the adaptive response to rifampicin (RFP)-induced cell injury. In this study, we found RFP decreased the survival rate of HepG2 cells and increased the levels of lactate dehydrogenase (LDH), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AKP), γ-glutamyl-transferase (γ-GT), total bilirubin (TBIL), direct bilirubin (DBIL), indirect bilirubin (IBIL), total bile acid (TBA) and adenosine triphosphate (ATP) in the cell culture supernatants in both a concentration- and a time-dependent manner. RFP increased the expression levels of bile acid transporter proteins and mRNAs, such as bile salt export pump (BSEP), multidrug resistance protein 1 (MDR1), multidrug resistance-associated protein 2 (MRP2), Na+/taurocholate cotransporter (NTCP), organic anion transporting protein 2 (OATP2), organic solute transporter ß (OSTß) and Nrf2. Following the transient knockdown of Nrf2 and treatment with RFP, the expression levels of the BSEP, MDR1, MRP2, NTCP, OATP2 and OSTß proteins and mRNAs were decreased to different degrees. Moreover, the cell survival was decreased, whereas the LDH level in the cell culture supernatant was increased. Overexpression of the Nrf2 gene produced the opposite effects. Treatment with tauroursodeoxycholic acid (TUDCA) increased the expression levels of the bile acid transporters and Nrf2, decreased the expression levels of glucose-regulated protein 78 (GRP78), PKR-like ER kinase (PERK), activating transcription factor 4 (ATF4), and C/EBP-homologous protein (CHOP), and inhibited RFP-induced oxidative stress. Moreover, TUDCA reduced cell apoptosis, increased cell survival and decreased the levels of LDH, ALT, AST, AKP, γ-GT, TBIL, DBIL, IBIL, TBA and ATP in the cell culture supernatant. Therefore, TUDCA alleviates RFP-induced injury in HepG2 cells by enhancing bile acid transporters expression and the Nrf2-mediated adaptive response.