Reexamining the Kuleshov effect: Behavioral and neural evidence from authentic film experiments.

Film cognition explores the influence of cinematic elements, such as editing and film color, on viewers' perception. The Kuleshov effect, a famous example of how editing influences viewers' emotional perception, was initially proposed to support montage theory through the Kuleshov experiment. This effect, which has since been recognized as a manifestation of point-of-view (POV) editing practices, posits that the emotional interpretation of neutral facial expressions is influenced by the accompanying emotional scene in a face-scene-face sequence. However, concerns persist regarding the validity of previous studies, often employing inauthentic film materials like static images, leaving the question of its existence in authentic films unanswered. This study addresses these concerns by utilizing authentic films in two experiments. In Experiment 1, multiple film clips were captured under the guidance of a professional film director and seamlessly integrated into authentic film sequences. 59 participants viewed these face-scene-face film sequences and were tasked with rating the valence and emotional intensity of neutral faces. The findings revealed that the accompanying fearful or happy scenes significantly influence the interpretation of emotion on neutral faces, eliciting perceptions of negative or positive emotions from the neutral face. These results affirm the existence of the Kuleshov effect within authentic films. In Experiment 2, 31 participants rated the valence and arousal of neutral faces while undergoing functional magnetic resonance imaging (fMRI). The behavioral results confirm the Kuleshov effect in the MRI scanner, while the neural data identify neural correlates that support its existence at the neural level. These correlates include the cuneus, precuneus, hippocampus, parahippocampal gyrus, post cingulate gyrus, orbitofrontal cortex, fusiform gyrus, and insula. These findings also underscore the contextual framing inherent in the Kuleshov effect. Overall, the study integrates film theory and cognitive neuroscience experiments, providing robust evidence supporting the existence of the Kuleshov effect through both subjective ratings and objective neuroimaging measurements. This research also contributes to a deeper understanding of the impact of film editing on viewers' emotional perception from the contemporary POV editing practices and neurocinematic perspective, advancing the knowledge of film cognition.

Irisin ameliorates UUO-induced renal interstitial fibrosis through TGF-ß1/periostin/MMP-2 signaling pathway.

Renal fibrosis is the most common pathway in progressive kidney diseases. The unilateral ureteral obstruction (UUO) model is used to induce progressive renal fibrosis. We evaluated the effects of irisin on renal interstitial fibrosis in UUO mice. The GSE121190, GSE36496, GSE42303, and GSE96101 datasets were downloaded from the Gene Expression Omnibus (GEO) database. In total, 656 differentially expressed genes (DEGs) were identified in normal and UUO mouse renal samples. Periostin and matrix metalloproteinase-2 (MMP-2) were selected to evaluate the effect of irisin on renal fibrosis in UUO mice. In UUO mice, irisin ameliorated renal function, decreased the expression of periostin and MMP-2, and attenuated epithelial-mesenchymal transition and extracellular matrix deposition in renal tissues. In HK-2 cells, irisin treatment markedly attenuated TGF-ß1-induced expression of periostin and MMP-2. Irisin treatment also inhibited TGF-ß1-induced epithelial-mesenchymal transition, extracellular matrix formation, and inflammatory responses. These protective effects of irisin were abolished by the overexpression of periostin and MMP-2. In summary, irisin treatment can improve UUO-induced renal interstitial fibrosis through the TGF-ß1/periostin/MMP-2 signaling pathway, suggesting that irisin may be used for the treatment of renal interstitial fibrosis.

Pan-Cancer Analysis and Experimental Validation of SOX4 as a Potential Diagnosis, Prognosis, and Immunotherapy Biomarker.

INTRODUCTION: SOX4 plays an important role in tumorigenesis and cancer progression. The role of SOX4 in pan-cancer and its underlying molecular mechanism in liver hepatocellular carcinoma (LIHC) are not fully understood. In this study, a comprehensive analysis and experimental validation were performed to explore the function of SOX4 across tumor types. METHODS: Raw data in regard to SOX4 expression in malignant tumors were downloaded from the TCGA and GTEx databases. The expression levels, prognostic values, genetic mutation, and DNA promoter methylation of SOX4 across tumor types were explored via systematic bioinformatics analysis. The ceRNA regulatory network, immune characteristics, and prognostic models were analyzed in LIHC. Finally, we conducted in vitro experiments including Western blotting, cell proliferative assay, trypan blue staining, and fluorescence microscopy to further explore the function of SOX4 in LIHC. RESULTS: SOX4 expression was significantly upregulated in 24 tumor types. SOX4 expression level was strongly associated with unfavorable prognoses, genetic mutations, and DNA methylation levels across different tumor types. Especially in LIHC, LINC00152/hsa-miR-139-3p/SOX4 was identified as a crucial ceRNA network. Moreover, this study also provides insight into the roles of SOX4 expression in immune cell infiltration, macrophage polarization, immune subtype, molecular subtype, and immunomodulators, as well as the tumor immune microenvironment (TIME)-related prognosis, in LIHC. The study established six favorable prognostic models to predict LIHC prognosis based on the SOX4-associated genes. Finally, lenvatinib treatment can increase the expression of SOX4 in hepatocellular carcinoma cells and lead to drug resistance. Silencing SOX4 can effectively eliminate the drug resistance caused by lenvatinib treatment and inhibit the proliferation of cancer cells. CONCLUSIONS: This study highlights that SOX4 may serve as a promising therapeutic target for tumor treatment.

Ring1a protects against colitis through regulating mucosal immune system and colonic microbial ecology.

Inflammatory bowel disease (IBD) represents a prominent chronic immune-mediated inflammatory disorder, yet its etiology remains poorly comprehended, encompassing intricate interactions between genetics, immunity, and the gut microbiome. This study uncovers a novel colitis-associated risk gene, namely Ring1a, which regulates the mucosal immune response and intestinal microbiota. Ring1a deficiency exacerbates colitis by impairing the immune system. Concomitantly, Ring1a deficiency led to a Prevotella genus-dominated pathogenic microenvironment, which can be horizontally transmitted to co-housed wild type (WT) mice, consequently intensifying dextran sodium sulfate (DSS)-induced colitis. Furthermore, we identified a potential mechanism linking the altered microbiota in Ring1aKO mice to decreased levels of IgA, and we demonstrated that metronidazole administration could ameliorate colitis progression in Ring1aKO mice, likely by reducing the abundance of the Prevotella genus. We also elucidated the immune landscape of DSS colitis and revealed the disruption of intestinal immune homeostasis associated with Ring1a deficiency. Collectively, these findings highlight Ring1a as a prospective candidate risk gene for colitis and suggest metronidazole as a potential therapeutic option for clinically managing Prevotella genus-dominated colitis.

We found that PcG protein Ring1a could be a new risk gene for colitis. Ring1a deficiency causes aggravated colitis by regulating the mucosal immune system and colonic microbial ecology.

Global research trends in unilateral ureteral obstruction-induced renal fibrosis: A bibliometric and visualized study.

BACKGROUND: Renal fibrosis is considered the pathway from almost all chronic kidney diseases (CKD) to end-stage renal diseases. The unilateral ureteral obstruction (UUO) model is a well-established experimental animal model to simulate renal fibrosis associated with obstructive nephropathy in an accelerated manner. In this study, in order to understand the development trends of research on UUO-induced renal fibrosis between 2005 and 2022 and predict prospects, we conducted a comprehensive bibliometric and visualized study using Web of Science (WoS). METHODS: The articles regarding UUO-induced renal fibrosis were culled from the "Core Collection" of the WoS database. VOSviewer software and the R-Bibliometrix Package were used in visual analysis of countries/regions, journals, authors, keywords, institutions, and highly cited articles in this field. RESULTS: The number of articles regarding UUO-induced renal fibrosis has obviously increased annually. China had the largest number of publications in this field. The most frequently used keywords were "inflammation," "transforming growth factor-beta1," "oxigative stress," "smad3," "beta-catenin," and "autophagy." Am J Physiol-Renal was the leading journal. The most highly influential documents were published by Higgins DF and his colleagues, with 46 local citations and 749 global citations. The leading institution was Nanjing Medical University. Furthermore, Zhang Y. was the author who contributed most to this field. CONCLUSION: Our results suggest that the molecular mechanism of UUO-induced renal fibrosis remains a research hot topic, especially on the inflammatory response and oxidative stress, and international cooperation is expected to expand and deepen in the future.

Classification and function of γδT cells and its research progress in anti-glioblastoma.

Human peripheral blood T lymphocytes are classified into alpha-beta T (αßΤ) cells and gamma-delta T (γδΤ) cells based on the difference in T cell receptors (TCRs). αßT cells are crucial for the acquired immune response, while Î³Î´Τ cells, though only a small subset, can recognize antigenic substances. These antigens do not need to be processed and presented and are not restricted by MHC. This distinguishes Î³Î´Τ cells from αßT cells and highlights their distinct role in innate immunity. Despite their small number, Î³Î´Τ cells hold significant significance in anti-tumor, anti-infection and immune regulation. Glioblastoma (GBM) represents one of the most prevalent malignant tumors within the central nervous system (CNS). Surgical resection alone proves to be an ineffective method for curing this type of cancer. Even with the combination of surgical resection, radiotherapy, and chemotherapy, the prognosis of some individuals with glioblastoma is still poor, and the recurrence rate is high. In this research, the classification, biological, and immunological functions of γδT cells and their research progress in anti-glioblastoma were reviewed.

Realizing High-Efficiency Orange-Red Thermally Activated Delayed Fluorescence Materials through the Construction of Intramolecular Noncovalent Interactions.

The development of highly efficient orange and red thermally activated delayed fluorescence (TADF) materials for constructing full-color and white organic light-emitting diodes (OLEDs) remains insufficient because of the formidable challenges in molecular design, such as the severe radiationless decay and the intrinsic trade-off between the efficiencies of radiative decay and reverse intersystem crossing (RISC). Herein, we design two high-efficiency orange and orange-red TADF molecules by constructing intermolecular noncovalent interactions. This strategy could not only ensure high emission efficiency via suppression of the nonradiative relaxation and enhancement of the radiative transition but also create intermediate triplet excited states to ensure the RISC process. Both emitters exhibit typical TADF characteristics, with a fast radiative rate and a low nonradiative rate. Photoluminescence quantum yields (PLQYs) of the orange (TPA-PT) and orange-red (DMAC-PT) materials reach up to 94 and 87%, respectively. Benefiting from the excellent photophysical properties and stability, OLEDs based on these TADF emitters realize orange to orange-red electroluminescence with high external quantum efficiencies reaching 26.2%. The current study demonstrates that the introduction of intermolecular noncovalent interactions is a feasible strategy for designing highly efficient orange to red TADF materials.

Irisin Ameliorates Renal Tubulointerstitial Fibrosis by Regulating the Smad4/ß-Catenin Pathway in Diabetic Mice.

Background: The primary pathophysiology of diabetic kidney disease (DKD) is tubulointerstitial fibrosis (TIF), and an essential contributing element is excessive extracellular matrix deposition. Irisin is a polypeptide formed by splitting fibronectin type III domain containing 5 (FNDC5), which participates in a number of physiological and pathological processes. Methods: The purpose of this article is to examine irisin's function in DKD and analyze both its in vitro and in vivo effects. The Gene Expression Omnibus (GEO) database was used to download GSE30122, GSE104954, and GSE99325. Analysis of renal tubule samples from nondiabetic and diabetic mice identified 94 differentially expressed genes (DEGs). The transforming growth factor beta receptor 2 (TGFBR2), irisin, and TGF-ß1 were utilized as DEGs to examine the impact of irisin on TIF in diabetic kidney tissue, according to the datasets retrieved from the GEO database and Nephroseq database. Additionally, the therapeutic impact of irisin was also examined using Western blot, RT-qPCR, immunofluorescence, immunohistochemistry, and kits for detecting mouse biochemical indices. Results: In vitro, the findings demonstrated that irisin not only down-regulated the expression of Smad4 and ß-catenin but also reduced the expression of proteins linked to fibrosis, the epithelial-mesenchymal transition (EMT), and mitochondrial dysfunction in HK-2 cells maintained in high glucose (HG) environment. In vivo, overexpressed FNDC5 plasmid was injected into diabetic mice to enhance its expression. Our studies found that overexpressed FNDC5 plasmid not only reversed the biochemical parameters and renal morphological characteristics of diabetic mice but also alleviated EMT and TIF by inhibiting Smad4/ß-catenin signaling pathway. Conclusion: The above experimental results revealed that irisin could reduce TIF in diabetic mice via regulating the Smad4/ß-catenin pathway.

Effects of particle size on the structure, cooking quality and anthocyanin diffusion of purple sweet potato noodles.

The effects of different particle sizes of purple sweet potato flour (PSPF) on the structure and quality of noodles and the diffusion kinetics of anthocyanins during cooking were studied. As the particle size of the PSPF decreased (from 269 to 66 µm), the adverse effects of the addition of PSPF on the quality of noodles were reduced. The smaller particle size of PSPF was beneficial for the secondary structure orderliness and the tighter microstructure of PSP noodles. The diffusion of anthocyanins in noodles to the soup during cooking could be fitted well with Fick's second law, and diffusion coefficients were in the range of 8.3248-14.0893 × 10-9 m2/s. The noodles with 15% 66 µm PSPF showed the best cooking properties, the highest sensory score, the highest anthocyanin retention ability and a compact and orderly microstructure. Thus, they could be considered as noodles rich in anthocyanins for commercial application.

Serological reactivity of inactivated SARS-CoV-2 vaccine based on an S-RBD neutralizing antibody assay.

OBJECTIVES: This study aimed to explore the regularity of S-RBD domain antibody reactivity after immunization with inactivated SARS-CoV-2 vaccine and evaluate the effect of this vaccine on the immune response. DESIGN OR METHODS: Venous blood samples were collected from 1156 healthcare workers who participated in the phase III clinical trial of the SARS-CoV-2 inactivated vaccine. The S-RBD domain antibody levels in the serum were detected by ELISA 14 days after the first and second active immunization, respectively. RESULTS: The positive rates after inoculation of the first and second vaccination of S-RBD domain antibody against SARS-CoV-2 were 28.03% and 86.76%, respectively. The mean inhibition rate of S-RBD domain antibody against positive samples was 57.18 ± 18.87% after the second vaccination at 14 days. Sex and age had no effects on the positive rate. The positive rate was decreased in the high BMI group. Single-factor logistic analysis showed that there was no significant correlation between age and positive rate. BMI was negatively correlated with the positive rate. CONCLUSIONS: After 2 immunizations, the positive rate of SARS-CoV-2 S-RBD domain antibody was high, and the vaccine had good immunogenicity. The improvement of the immune strategy should focus on the effects of BMI and other factors.

Biological characteristics of γδT cells and application in tumor immunotherapy.

Human γδT cells are a special immune cell type which exist in small quantities in the body, do not require processing and presentation for antigen recognition, and have non-major histocompatibility complex (MHC)-restricted immune response. They play an important role in the body's anti-tumor, anti-infection, immune regulation, immune surveillance and maintenance of immune tolerance. This article reviews the generation and development of human γδT cells, genetic characteristics, classification, recognition and role of antigens, and research progress in tumor immunotherapy.

Detection of nucleated red blood cells using the Mindray BC-6800Plus hematology analyzer: a clinical performance evaluation.

BACKGROUND: With the development of instrument technology, the functions and detection methods of automatic blood cell analyzers have become more complex. To ensure optimal clinical applicability, it is crucial to select an automatic blood cell analyzer with excellent clinical detection performance. This study evaluated the latest Mindray BC-6800Plus automatic blood cell analyzer and assessed its performance in the detection of nucleated red blood cells (NRBCs). METHODS: A total of 490 clinical blood samples were used to assess the performance of the instrument, including parameters such as precision, linearity, conformity rate of manual microscopic examination, carryover, and limit of quantitation. RESULTS: The instrument showed a small carryover (≤0.02) and excellent linearity (R2≥0.9986). The reproducibility of the sample tests was satisfactory, and the coefficient of variation (CV) of the test results [0.98-1.72% and 0.62-6.97% for white blood cells (WBCs) and NRBCs, respectively] were significantly lower than that declared by the manufacturer (2.5% and 20% for WBCs and NRBCs, respectively). Thus, the BC-6800Plus satisfies the requirements of clinical testing. Two separate Mindray BC-6800Plus machines were tested and found to be in good agreement with each other and with manual microscopy methods. Furthermore, WBC and NRBC counts were highly consistent with results obtained using the XN-9100 blood analyzer. CONCLUSIONS: The Mindray BC-6800Plus is an excellent analyzer that can provide timely and accurate reports for clinical laboratory detection of NRBC.

Effects of influenza vaccination on clinical outcomes of chronic obstructive pulmonary disease: A systematic review and meta-analysis.

PURPOSE: Influenza is a threat to patients with chronic obstructive pulmonary disease (COPD), influenza vaccination help to reduce incidence of influenza infection, however, whether it is beneficial to COPD patients in clinical outcomes lacks for evidence due to limited studies and participations. METHODS: We searched PubMed, Embase, Cochrane Library, China National Knowledge Infrastructure (CNKI) and China Science and Technology Journal Database (CSTJ) to retrieve eligible studies regardless of study design published before August 2020, and conducted meta-analysis with odds ratio (OR) and mean difference (MD). The quality of included studies and pooled evidences were assessed. Narrative summaries were provided where data were insufficient for meta-analysis. RESULTS: 2831 COPD patients were included, the pooled results showed that influenza vaccination reduced the exacerbations (P = 0.0001) and trends of hospitalizations (P = 0.09) in COPD patients. Further subgroup analysis showed that the reduction of exacerbations and hospitalizations were significant in patients with FEV1<50 % predicted (P = 0.01 and P < 0.0001 respectively), but not in those with FEV1≥50 % predicted (P = 0.23 and P = 0.76 respectively). No significant effect of influenza vaccination on all-cause mortality was observed. CONCLUSIONS: Our findings support a protective role of influenza vaccination in COPD patients, a yearly influenza vaccination should be strongly recommended for all COPD patients, especially those with severe airflow limitation, to reduce possible influenza infection, and thus associated exacerbations and hospitalizations.

Procyanidin B2 improves endothelial progenitor cell function and promotes wound healing in diabetic mice via activating Nrf2.

One of the major reasons for the delayed wound healing in diabetes is the dysfunction of endothelial progenitor cells (EPCs) induced by hyperglycaemia. Improvement of EPC function may be a potential strategy for accelerating wound healing in diabetes. Procyanidin B2 (PCB2) is one of the major components of procyanidins, which exhibits a variety of potent pharmacological activities. However, the effects of PCB2 on EPC function and diabetic wound repair remain elusive. We evaluated the protective effects of PCB2 in EPCs with high glucose (HG) treatment and in a diabetic wound healing model. EPCs derived from human umbilical cord blood were treated with HG. The results showed that PCB2 significantly preserved the angiogenic function, survival and migration abilities of EPCs with HG treatment, and attenuated HG-induced oxidative stress of EPCs by scavenging excessive reactive oxygen species (ROS). A mechanistic study found the protective role of PCB2 is dependent on activating nuclear factor erythroid 2-related factor 2 (Nrf2). PCB2 increased the expression of Nrf2 and its downstream antioxidant genes to attenuate the oxidative stress induced by HG in EPCs, which were abolished by knockdown of Nrf2 expression. An in vivo study showed that intraperitoneal administration of PCB2 promoted wound healing and angiogenesis in diabetic mice, which was accompanied by a significant reduction in ROS level and an increase in circulating EPC number. Taken together, our results indicate that PCB2 treatment accelerates wound healing and increases angiogenesis in diabetic mice, which may be mediated by improving the mobilization and function of EPCs.

Identification of key gene modules and genes in colorectal cancer by co-expression analysis weighted gene co-expression network analysis.

Colorectal cancer (CRC) has been one of the most common malignancies worldwide, which tends to get worse for the growth and aging of the population and westernized lifestyle. However, there is no effective treatment due to the complexity of its etiology. Hence, the pathogenic mechanisms remain to be clearly defined. In the present study, we adopted an advanced analytical method-Weighted Gene Co-expression Network Analysis (WGCNA) to identify the key gene modules and hub genes associated with CRC. In total, five gene co-expression modules were highly associated with CRC, of which, one gene module correlated with CRC significantly positive (R = 0.88). Functional enrichment analysis of genes in primary gene module found metabolic pathways, which might be a potentially important pathway involved in CRC. Further, we identified and verified some hub genes positively correlated with CRC by using Cytoscape software and UALCAN databases, including PAICS, ATR, AASDHPPT, DDX18, NUP107 and TOMM6. The present study discovered key gene modules and hub genes associated with CRC, which provide references to understand the pathogenesis of CRC and may be novel candidate target genes of CRC.

Highly selective fluorimetric and colorimetric sensing of mercury(II) by exploiting the self-assembly-induced emission of 4-chlorothiophenol capped copper nanoclusters.

A highly stable copper nanoclusters (CuNC) carrying 4-chlorothiophenol as a protective ligand is described. They display self-assembly-induced emission with excitation/emission maxima at 330/605 nm even in neutral or alkaline aqueous environment. The fluorescence of these CuNC is quenched by Hg(II). Quenching is mainly ascribed to the formation of a complex formed via Hg-S bonding between the Hg(II) ions and the ligand. This destroys the ordered architectures of the assembled CuNC. The assay enables Hg(II) to be determined with good sensitivity and a linear response ranging from 1 to 500 nM Hg(II) with a 0.3 nM limit of detection. In addition, the method was implemented in a test strip (which undergoes a color change from red to blue) that can be used for visual determination of Hg(II) in complex environmental water samples. Graphical abstractNovel and highly selective fluorimetric and colorimetric methods have been designed for mercury(II) ions determination based on stable self-assembly-induced emission of copper nanoclusters.

The elevated CXCL5 levels in circulation are associated with lung function decline in COPD patients and cigarette smoking-induced mouse model of COPD.

Introduction: C-X-C motif chemokine 5 is primarily chemotactic for neutrophils and previously shown to increase in the bronchoalveolar lavage fluid of patients with chronic obstructive pulmonary disease. However, whether C-X-C motif chemokine 5 levels correlate with lung function decline in patients or mouse model of chronic obstructive pulmonary disease was not clear. Methods: The mouse model was induced by cigarette smoke exposure. Plasma/serum and bronchoalveolar lavage fluid were obtained from patients and mouse model of chronic obstructive pulmonary disease; C-X-C motif chemokine 5 levels were assessed and correlated with lung functions and granulocyte-colony stimulating factor levels, respectively. Results: The C-X-C motif chemokine 5 levels increased and correlated to granulocyte-colony stimulating factor levels in both plasma/serum and bronchoalveolar lavage fluid obtained from patients and mouse model of chronic obstructive pulmonary disease. Circulating levels of C-X-C motif chemokine 5 correlated to lung functions decline in patients and mouse model. Conclusions: Granulocyte-colony stimulating factor might coordinate with C-X-C motif chemokine 5 in the pathogenesis of neutrophilic inflammation in chronic obstructive pulmonary disease. Circulating C-X-C motif chemokine 5 might serve as a potential blood-based biomarker to add additional modest predictive value on the preliminary screening and diagnosis of chronic obstructive pulmonary disease. Key messages Circulating C-X-C motif chemokine 5 might serve as a potential blood-based biomarker to add additional modest predictive value on the preliminary screening and diagnosis of COPD. Granulocyte-colony stimulating factor might coordinate with C-X-C motif chemokine 5 in the pathogenesis of neutrophilic inflammation in chronic obstructive pulmonary disease.

Natural plant extract tubeimoside I promotes apoptosis-mediated cell death in cultured human hepatoma (HepG2) cells.

Tubeimoside I (TBMS I), an extract from Chinese herbal medicine Bolbostemma paniculatum (MAXIM.) FRANQUET (Cucurbitaceae) has been shown as a potent anti-tumor agent for a variety of human cancers, but yet to be evaluated for hepatoma that is highly prevalent in Eastern Asian countries including China. Here, we examined in vitro the cytotoxic effects of TBMS I on human hepatoma (HepG2) and normal liver (L-02) cell lines. We also investigated TBMS I-induced molecular events related to apoptosis in HepG2 cells. The results show that TBMS I inhibited the proliferation of both HepG2 and L-02 cells in a dose- and time-dependent manner, but HepG2 cells appeared more sensitive to the agent. When exposed to TBMS I for 24, 48 and 72 h, IC50 for HepG2 cells versus L-02 cells were 15.5 vs. 23.1, 11.7 vs. 16.2, 9.2 vs. 13.1 (µM, p<0.01), respectively. TBMS I induced cell shrinkage, nuclear condensation and fragmentation, cell cycle arrest at the G2/M phase, mitochondrial membrane disruption, release of cytochrome c from the mitochondria, activation of caspase 3 and 9, and shifting Bax/Bcl-2 ratio from being anti-apoptotic to pro-apoptotic, all indicative of initiation and progression of apoptosis involving mitochondrial dysfunction. Taken together, these results indicate for the first time that TBMS I potently inhibited growth in HepG2 cells by mediating a cascade of apoptosis signaling pathways. Considering its sensitivity of HepG2 cells, preferential distribution in the liver and natural product origin, TBMS I therefore may have a great potential as a chemotherapeutic drug candidate for hepatoma.

Natural plant extract tubeimoside I induces cytotoxicity via the mitochondrial pathway in human normal liver cells.

Tubeimoside I (TBMS I) is a natural compound extracted from Bolbostemma paniculatum (Maxim.) Franquet (Cucurbitaceae), a traditional Chinese herbal medicine widely used for the treatment of inflammation. Recently, it has been suggested that TBMS I may be a potent anticancer agent for a variety of human cancers. However, TBMS I is known to distribute preferentially in the liver, and thus may harm normal liver cells if it is delivered systemically for cancer treatment. This safety concern warrants careful evaluation of the hepatotoxicity of TBMS I to normal liver cells, which to date has not been carried out. Here, we report the cytotoxic effects of TBMS I on one type of normal liver cells (L-02 cells), and the associated molecular events as underlying mechanisms. Cultured human normal liver L-02 cells were treated with TBMS I at concentrations of 0, 15 and 30 µM for 24, 48 and 72 h, respectively. Subsequently, the cell survival rate was evaluated by the MTT dye method, and several key molecular events associated with apoptosis were assayed, including mitochondrial depolarization, release of cytochrome c (cyt-c), activation of caspases, and the balance between Bax and Bcl-2 protein expression. Our results indicate that TBMS I inhibited the proliferation of L-02 cells in a dose- and time-dependent manner. The TBMS I-induced growth inhibition of L-02 cells was accompanied by the collapse of mitochondrial membrane potential, release of cyt-c from the mitochondria to the cytosol, activation of caspase-9 and -3, decrease of anti-apoptotic protein Bcl-2 levels and increase of the pro-apoptotic protein Bax levels, all indicative of apoptosis through the mitochondrial pathway. Taken together, these results confirm that TBMS I has a significant apoptotic effect on normal liver L-02 cells, which may be significant to its clinical applications.