Repositioning VU-0365114 as a novel microtubule-destabilizing agent for treating cancer and overcoming drug resistance.

Microtubule-targeting agents represent one of the most successful classes of anticancer agents. However, the development of drug resistance and the appearance of adverse effects hamper their clinical implementation. Novel microtubule-targeting agents without such limitations are urgently needed. By employing a gene expression-based drug repositioning strategy, this study identifies VU-0365114, originally synthesized as a positive allosteric modulator of human muscarinic acetylcholine receptor M5 (M5 mAChR), as a novel type of tubulin inhibitor by destabilizing microtubules. VU-0365114 exhibits a broad-spectrum in vitro anticancer activity, especially in colorectal cancer cells. A tumor xenograft study in nude mice shows that VU-0365114 slowed the in vivo colorectal tumor growth. The anticancer activity of VU-0365114 is not related to its original target, M5 mAChR. In addition, VU-0365114 does not serve as a substrate of multidrug resistance (MDR) proteins, and thus, it can overcome MDR. Furthermore, a kinome analysis shows that VU-0365114 did not exhibit other significant off-target effects. Taken together, our study suggests that VU-0365114 primarily targets microtubules, offering potential for repurposing in cancer treatment, although more studies are needed before further drug development.

A Bifunctional Fluorescence Probe Based on AIE-ICT Strategy for Visual Detection of Cu2+/Co2+ in Complex Matrix.

A novel fluorescence chemical sensor-based probe 1-{[(E)-(2-aminophenyl)azanylidene]methyl}naphthalen-2-ol (AMN) was designed and synthesized, which performed a "naked eye" detection ability toward Cu2+ and Co2+ based on aggregation-induced emission (AIE) fluorescence strategy. It has sensitive detection ability for Cu2+ and Co2+. In addition, the color changed from yellow-green to orange under the sunlight, realizing the rapid identification of Cu2+/Co2+, which has the potential of on-site visual detection under the "naked eye". Moreover, different "on" and "off" fluorescence expressions were exhibited under excessive glutathione (GSH) in AMN-Cu2+ and AMN-Co2+ systems, which could be employed to distinguish Cu2+ from Co2+. The detection limits for Cu2+ and Co2+ were measured to be 8.29 × 10-8 M and 9.13 × 10-8 M, respectively. The binding mode of AMN was calculated to be 2:1 by Jobs' plot method analysis. Ultimately, the new fluorescence sensor was applied to detect Cu2+ and Co2+ in real samples (tap water, river water, and yellow croaker), and the results were satisfying. Therefore, this high-efficiency bifunctional chemical sensor platform based on "on-off" fluorescence detection will provide significant guidance for the advance development of single-molecule sensors for multi-ion detection.

Effect of Atropine 0.01% Eye Drops on the Difference in Refraction and Axial Length between Right and Left Eyes.

INTRODUCTION: This study sought to determine whether the application of 0.01% atropine eye drops could impact the disparity in refraction and axial length (AL) between the right and left eyes in Chinese children. METHODS: The study was designed as a double-blind, placebo-controlled randomized trial. A total of 220 children aged 6-12 years were recruited from the Beijing Tongren Hospital in Beijing, China. Participants were randomized in a 1:1 ratio and were prescribed 0.01% atropine or placebo eye drops to be administered once a night to both eyes for the duration of 1 year. The cycloplegic refraction and AL were recorded including baseline, 6 months, and again at the 12 months. RESULTS: After 1-year follow-up period, 76 (69%) and 83 (75%) subjects of the initial 220 participants were identified as the 0.01% atropine and placebo groups, respectively. The inter-ocular difference in spherical equivalent refraction (SER) and AL demonstrated stable values in the 0.01% atropine treatment group (SER: p = 0.590; AL: p = 0.322) analyzed after 1 year, but found a significant increase (SER: p < 0.001; AL: p = 0.001) in the placebo group. Furthermore, over 1 year, eyes with greater myopia in the atropine group exhibited slower myopia progression (0.45 ± 0.44 D) than the lesser myopic eye (0.56 ± 0.44 D) (p = 0.003). CONCLUSION: This study demonstrated that 0.01% atropine could maintain the inter-ocular SER and AL difference. And 0.01% atropine appeared to be more effective in delaying the progression of myopia in eyes with more myopia than in the less myopic eyes.

Myopia progression after cessation of low-dose atropine eyedrops treatment: A two-year randomized, double-masked, placebo-controlled, cross-over trial.

PURPOSE: The purpose of the study was to evaluate myopia progression and axial elongation after stopping 0.01% atropine eye drops through a 2-year cross-over study. METHODS: This study was a randomized, double-masked, placebo-controlled, cross-over trial in mainland China. 220 children aged 6-12 years with spherical equivalent range of -1.00 D to -6.00 D in both eyes were enrolled in Phase 1 for 1 year. Children who had completed the first year's follow-up continued in the second phase. In Phase 2, the placebo group was crossed over to the 0.01% atropine group (referred to as the 'placebo-atropine group'), and the 0.01% atropine group was crossed over to the placebo group (referred to as the 'atropine-placebo group'). All children underwent the examination of cycloplegic refraction and axial length at a 6-month interval. Only data from right eyes were included in analysis. RESULTS: One hundred thirty-three subjects completed 2 years of follow-up. In the first year, the mean myopia progression in atropine-placebo group was 0.21 ± 0.08 D slower than that in placebo-atropine group. After cross-over treatment, the mean myopia progression in atropine-placebo group was 0.22 ± 0.07D faster than that in placebo-atropine group in the second year. Over 2 years, the mean myopia progression was -1.26 ± 0.66D and -1.25 ± 0.70D in the atropine-placebo and placebo-atropine groups (p = 0.954). CONCLUSIONS: The difference in myopia progression between atropine-placebo group and placebo-atropine group in Phase 1 was similar to Phase 2 during the cross-over treatment. Through our cross-over trial, the results suggest that there is no rebound effect after using 0.01% atropine eye drops to prevent progression of myopia.

The effect of atropine 0.01% eyedrops on relative peripheral refraction in myopic children.

BACKGROUND: Relative peripheral refraction (RPR) is a significant factor that participates in myopic development. Here, we evaluated the effects of atropine 0.01% eyedrops, as an antimyopia drug, on RPR. METHODS: Seventy-three children were enrolled from a randomized, double-blinded, placebo-0.01% atropine eyedrops cross-over trial. The study group had used the placebo for one year and then crossed over to atropine 0.01% eyedrops for half a year. The control group had used 0.01% atropine for one year and then crossed over to placebo eyedrops for half a year. Central and horizontal peripheral refractions (15° and 30° at the temporal and nasal retina) were measured under non-cycloplegia and cycloplegia. RESULTS: No significant differences in age, gender, and central refraction were identified between the two groups (P > 0.05). Under non-cycloplegia, the control group showed significant relative hyperopia in the temporal 30° retina and the nasal retina (P = 0.031; P < 0.001; P < 0.001). In the study group, the relative hyperopia in the temporal 30° retina disappeared (P = 0.983). After cycloplegia, the control group had less myopia in central refractions and less hyperopia in temporal RPR (P < 0.001; P = 0.039; P < 0.001). The study group did not present significant changes in central refractions and temporal RPR (P = 0.122; P = 0.222; P = 0.475). CONCLUSIONS: For myopic children, atropine 0.01% eyedrops can alleviate relative hyperopia in the temporal retina and the hyperopic shift before cycloplegia. The effect might participate in myopia control.

Effect of 0.01% atropine eyedrops on intraocular pressure in schoolchildren: a randomized clinical trial.

AIM: To assess the effect of 0.01% atropine eye drops on intraocular pressure (IOP) in myopic children. METHODS: A placebo-controlled, double-masked, randomized study. Totally 220 children aged 6 to 12y with myopia ranging from -1.00 to -6.00 D in both eyes were enrolled. Children were randomized in a 1:1 ratio to either 0.01% atropine eye drops or a placebo group using generated random numbers. All participants underwent the examination of IOP and cycloplegic refraction at baseline, 6 and 12mo. The change of IOP and the proportion of subjects with increased IOP in atropine and placebo groups were compared. RESULTS: Of 220 children, 117 were boys (53.2%). A total of 159 (72.3%) participants completed the follow-up at the 1-year study. At baseline, the mean IOP was 15.74 mm Hg (95%CI, 15.13 to 16.34 mm Hg) for the 0.01% atropine group and 15.59 mm Hg (95%CI, 15.00 to 16.19 mm Hg) for placebo group (mean difference, 0.14 mm Hg; P=0.743) after adjusting for central corneal thickness at baseline. At one year follow-up, the mean change of IOP was 0.16 mm Hg (95%CI, -0.43 to 0.76 mm Hg) for the 0.01% atropine group and -0.11 mm Hg (95%CI, -0.71 to 0.50 mm Hg) for placebo group (mean difference, 0.27 mm Hg; P=0.525) after adjusting for central corneal thickness. The 51.4% of children have increased IOP in the 0.01% atropine group, compared with 45.9% in the placebo group (P=0.511). CONCLUSION: The 0.01% atropine eye drops do not significantly affect the risk of elevated IOP. It is relatively safer to use in the studies that try to minimize myopia progression. However, a further long-duration study is required to be validated.

Brain Activation Induced by Myopic and Hyperopic Defocus From Spectacles.

Purpose: To assess neural changes in perceptual effects induced by myopic defocus and hyperopic defocus stimuli in ametropic and emmetropic subjects using functional magnetic resonance imaging (fMRI). Methods: This study included 41 subjects with a mean age of 26.0 ± 2.9 years. The mean spherical equivalence refraction was -0.54 ± 0.51D in the emmetropic group and -3.57 ± 2.27D in the ametropic group. The subjects were instructed to view through full refractive correction, with values of +2.00D to induce myopic defocus state and -2.00D to induce hyperopic defocus state. This was carried over in three random sessions. Arterial spin labeling (ASL) perfusion was measured using fMRI to obtain quantified regional cerebral blood flow (rCBF). Behavioral tests including distant visual acuity (VA) and contrast sensitivity (CS), were measured every 5 min for 30 min. Results: Myopic defocus induced significantly greater rCBF increase in four cerebral regions compared with full correction: right precentral gyrus, right superior temporal gyrus, left inferior parietal lobule, and left middle temporal gyrus (P < 0.001). The differences were less significant in low myopes than emmetropes. In the hyperopic defocus session, the increased responses of rCBF were only observed in the right and left precentral gyrus. Myopic defocused VA and CS improved significantly within 5 min and reached a plateau shortly after. Conclusion: This study revealed that myopic defocus stimuli can significantly increase blood perfusion in visual attention-related cerebral regions, which suggests a potential direction for future investigation on the relationship between retinal defocus and its neural consequences.

Mesenchymal stem cells: Biological characteristics and application in disease therapy.

Mesenchymal stem cells (MSCs) are multipotent stem cells. In addition to the capacity for self-renewal and multipotential differentiation, MSCs also have the following characteristics. MSCs can exert immunomodulatory functions through interaction with innate or adaptive immune cells, MSCs with poor immunogenicity can be used for allogeneic transplantation, and MSCs can "home" to inflammation and tumour sites. Based on these biological properties, MSCs demonstrate broad clinical application prospects in the treatment of tissue injury, autoimmune diseases, transplantation, cancer and other inflammation-related diseases. In this review we describe the biological characteristics of MSCs and discuss the research advances of MSCs in regenerative medicine, immunomodulation, oncology, and COVID-19, to fully understand the range of diseases in which MSC therapy may be beneficial.

Effect of reading with a mobile phone and text on accommodation in young adults.

PURPOSE: To investigate the effects of reading with mobile phone versus text on accommodation accuracy and near work-induced transient myopia (NITM) and its subsequent decay during near reading in young adults with mild to moderate myopia. METHODS: The refractions of 31 young adults were measured with an open-field autorefractor (WAM-5500, Grand Seiko) for two reading tasks with a mobile phone and text at 33 cm. The mean age of the young adults was 24.35 ± 1.80 years. The baseline refractive aspects were determined clinically with full distance refractive correction in place. The initial NITM and its decay time and accommodative lag were assessed objectively immediately after binocularly viewing a mobile phone or text for 40 min. RESULTS: The mean ± standard deviation (SD) initial NITM magnitude was greater for reading with text (0.23 ± 0.26 D) than for reading with mobile phone (0.12 ± 0.17 D), but there was no significant difference between the two reading tasks (p = 0.082). The decay time (median, first quartile, and third quartile) was 60 s (16, 154) and 70 s (32, 180) in the phone task and text task groups, respectively. There was also no significant difference in the decay time between the two reading types in general (p = 0.294). The accommodative lags of text tasks and mobile phones tasks were equivalent (1.27 ± 0.52 D vs 1.31 ± 0.64 D, p = 0.792). CONCLUSION: There were no significant differences in accommodative lags and the initial NITM and its decay time between reading with a mobile phone and text in young adults.

Safety and Efficacy of Low-Dose Atropine Eyedrops for the Treatment of Myopia Progression in Chinese Children: A Randomized Clinical Trial.

Importance: Because studies have suggested that atropine might slow the progression of myopia in children, randomized clinical trials are warranted to understand this potential causal relationship. Objective: To evaluate the efficacy and safety of atropine, 0.01%, eyedrops on slowing myopia progression and axial elongation in Chinese children. Design, Setting, and Participants: This was a randomized, placebo-controlled, double-masked study. A total of 220 children aged 6 to 12 years with myopia of -1.00 D to -6.00 D in both eyes were enrolled between April 2018 and July 2018 at Beijing Tongren Hospital, Beijing, China. Cycloplegic refraction and axial length were measured at baseline, 6 months, and 12 months. Adverse events were also recorded. Interventions: Patients were randomly assigned in a 1:1 ratio to atropine, 0.01%, or placebo groups to be administered once nightly to both eyes for 1 year. Main Outcomes and Measures: Mean changes and percentage differences in myopia progression and axial elongation between atropine, 0.01%, or placebo groups. Results: Of 220 participants, 103 were girls (46.8%), and the mean (SD) age was 9.64 (1.68) years. The mean (SD) baseline refractive error and axial length were -2.58 (1.39) D and 24.59 (0.87) mm. Follow-up at 1 year included 76 children (69%) and 83 children (75%) allocated into the atropine, 0.01%, and placebo groups, respectively, when mean myopia progression was -0.49 (0.42) D and -0.76 (0.50) D in the atropine, 0.01%, and placebo groups (mean difference, 0.26 D; 95% CI, 0.12-0.41 D; P < .001), with a relative reduction of 34.2% in myopia progression. The mean (SD) axial elongation in the atropine, 0.01%, group was 0.32 (0.19) mm compared with 0.41 (0.19) mm in the placebo group (mean difference, 0.09 mm; 95% CI, 0.03-0.15 mm; P = .004), with relative reduction of 22.0% in axial elongation. Fifty-one percent and 13.2% of children progressed by at least 0.50 D and 1.00 D in the atropine, 0.01%, group, compared with 69.9% and 34.9% in the placebo group. No serious adverse events related to atropine were reported. Conclusions and Relevance: While the clinical relevance of the results cannot be determined from this trial, these 1-year results, limited by approximately 70% follow-up, suggest that atropine, 0.01%, eyedrops can slow myopia progression and axial elongation in children and warrant future studies to determine longer-term results and potential effects on slowing sight-threatening pathologic changes later in life. Trial Registration: http://www.chictr.org.cn Identifier: ChiCTR-IOR-17013898.

Sequential Interferon ß-Cisplatin Treatment Enhances the Surface Exposure of Calreticulin in Cancer Cells via an Interferon Regulatory Factor 1-Dependent Manner.

Immunogenic cell death (ICD) refers to a unique form of cell death that activates an adaptive immune response against dead-cell-associated antigens. Accumulating evidence indicates that the efficacy of conventional anticancer agents relies on not only their direct cytostatic/cytotoxic effects but also the activation of antitumor ICD. Common anticancer ICD inducers include certain chemotherapeutic agents (such as anthracyclines, oxaliplatin, and bortezomib), radiotherapy, photodynamic therapy (PDT), and oncolytic virotherapies. However, most chemotherapeutic reagents are inefficient or fail to trigger ICD. Therefore, better understanding on the molecular determinants of chemotherapy-induced ICD will help in the development of more efficient combinational anticancer strategies through converting non- or relatively weak ICD inducers into bona fide ICD inducers. In this study, we found that sequential, but not concurrent, treatment of cancer cells with interferon ß (IFNß), a type I IFN, and cisplatin (an inefficient ICD inducer) can enhance the expression of ICD biomarkers in cancer cells, including surface translocation of an endoplasmic reticulum (ER) chaperone, calreticulin (CRT), and phosphorylation of the eukaryotic translation initiation factor alpha (eIF2α). These results suggest that exogenous IFNß may activate molecular determinants that convert cisplatin into an ICD inducer. Further bioinformatics and in vitro experimental analyses found that interferon regulatory factor 1 (IRF1) acted as an essential mediator of surface CRT exposure by sequential IFNß-cisplatin combination. Our findings not only help to design more effective combinational anticancer therapy using IFNß and cisplatin, but also provide a novel insight into the role of IRF1 in connecting the type I IFN responses and ICD.

Visual Impairment and Spectacle Use in University Students in Central China: The Anyang University Students Eye Study.

PURPOSE: To investigate the prevalence and associations of visual impairment and spectacle use in university students in central China. DESIGN: Cross-sectional study. METHODS: This study included students aged 16-26 years in China. Study subjects from 2 universities underwent distance visual acuity (VA) assessment in both eyes with a logarithm of the minimum angle of resolution chart and their refractions were measured by cycloplegic autorefraction. Blindness was defined as presenting VA less than three-sixtieth in the better eye (World Health Organization definition), and visual impairment was defined as presenting VA less than six-twelfths. RESULTS: Overall, 9710 undergraduates were enumerated, 7704 (79.3%) subjects were included in this study. The prevalence of uncorrected VA less than six-twelfths and less than three-sixtieth in the better eye were 69.9% and 0.9%, respectively. Only 77.0% (4148/5388) of subjects with uncorrected VA in the better eye of less than six-twelfths wore glasses. For presenting VA, the prevalence of mild (VA <6/12 to 6/18), moderate (VA <6/18 to 6/60), and severe (VA <6/60 to 3/60) visual impairment was 6.3%, 11.2%, and 0.7%, respectively. Overall, 71.7% (4300/6001) of students with myopia (spherical equivalent ≤-0.5 diopters) wore spectacles. In multiple logistic regression analysis, visual impairment was associated with female sex (P < .001) and lower year level of education (P = .006) when presenting with VA. CONCLUSIONS: This study has documented a relatively high prevalence of visual impairment and relatively low spectacle coverage in Chinese university students. Given the potential impact of visual impairment, target education and accessible refraction services are highly important to solve the problem.

Refractive Errors in University Students in Central China: The Anyang University Students Eye Study.

Purpose: To assess the prevalence of refractive errors and associated factors in university students in urban areas of Anyang, Central China. Methods: This is a cross-sectional university-based study of 16- to 26-year-old students in China. Subjects from two universities were invited to undergo a comprehensive eye examination. Cycloplegic refraction was acquired by autorefractor with two drops of 1% cyclopentolate. The prevalence of myopia, high myopia, hyperopia, astigmatism, and anisometropia was calculated. Only data from right eyes were included in analysis. Results: A total of 7732 eligible subjects were included, with an average age of 20.2 ± 1.4 years. Overall, the mean spherical equivalent (SE) was -2.92 ± 2.48 diopters (D). The prevalence of myopia (SE ≤ -0.50 D), emmetropia, and hyperopia (SE ≥ +0.50 D) was 83.2%, 9.5%, and 7.3%, respectively. Female sex (OR = 1.542; P < 0.001) and science and engineering students (OR = 1.219; P = 0.004) were more likely to be myopic. The prevalence of high myopia, defined using SE ≤ -5.0 D, ≤ -6.0 D, and ≤ -10.0 D, respectively, occurred in 20.2%, 11.1 %, and 0.5%. High myopia (SE ≤ -6.0 D) was statistically associated with female sex (OR = 1.202; P = 0.029) and younger age (OR = 0.896; P = 0.001). The prevalence of astigmatism (cylinder of ≤ -0.75 D) was 28.8%. Astigmatism was associated with male sex (OR = 0.824; P = 0.001) and younger age (OR = 0.925; P = 0.001). Conclusions: A prevalence of 83.2% for myopia and 11.1% for high myopia (SE ≤ -6.0 D) was found in central Chinese university students. In the future, this generation of university students may encounter long-term, vision-threatening effects, especially pathologic myopia.

Cycloplegic refraction by 1% cyclopentolate in young adults: is it the gold standard? The Anyang University Students Eye Study (AUSES).

AIMS: To document the difference between non-cycloplegic and cycloplegic refraction and explore its associated factors in Chinese young adults. METHODS: A school-based study including 7971 undergraduates was conducted in Anyang, Henan Province, China. Cycloplegia was achieved with two drops of 1% cyclopentolate and 1 drop of Mydrin P (Tropicamide 0.5%, phenylephrine HCl 0.5%) with a 5 min interval. Non-cycloplegic and cycloplegic refractions were measured by an autorefractor. A paired-sample t-test and Spearman correlation analysis were used for analysis with data from only the right eyes included.  RESULTS: Of the 7971 students examined, 7793 (97.8%) with complete data were included, aging 20.2±1.5 years. Male students accounted for 36.8%. Overall, there was a significant difference between non-cycloplegic and cycloplegic SE (spherical equivalent) of 0.83±0.81D (p<0.01). The difference was 1.80±1.11D, 1.26±0.93D and 0.69±0.69D for those with cycloplegic hyperopia, emmetropia and myopia, respectively (p<0.01 for all). Those with a hyperopic shift less than 0.25D and 0.5D accounted for 11.1% and 34.1%, respectively. A significant relationship was found between difference in SE and cycloplegic refraction (r=0.33, b=0.11, p<0.01). Without cycloplegia, prevalence of hyperopia and emmetropia would be underestimated by 6.2% (1.0% vs 7.2%) and 5.7% (3.8% vs 9.5%), respectively, with prevalence of myopia and high myopia overestimated by 12.1% (95.3% vs 83.2%) and 6.1% (17.2% vs 11.1%).  CONCLUSION: Lack of cycloplegia will lead to significant misclassification of myopia, emmetropia and hyperopia in Chinese young adults. Cycloplegia is therefore essential for this age-group in epidemiological studies.

The Chinese Herbal Mixture Tien-Hsien Liquid Augments the Anticancer Immunity in Tumor Cell-Vaccinated Mice.

BACKGROUND: The Chinese herbal mixture, Tien-Hsien liquid (THL), has been used as an anticancer dietary supplement for more than 20 years. Our previous studies have shown that THL can modulate immune responseand inhibit tumor growth. In this study, we further evaluated the effect of THL on anticancer immune response in mice vaccinated with γ-ray-irradiated tumor cells. METHODS: The antitumor effect of THL was determined in mice vaccinated with low-tumorigenic CT-26-low colon cancer cells or γ-ray-irradiated high-tumorigenic CT-26-high colon cancer cells. The number of natural killer (NK) cells and T lymphocytes in the spleen was analyzed by flow cytometry. The tumor-killing activities of NK cells and cytotoxic T lymphocytes (CTLs) were analyzed by flow cytometry using YAC-1 and CT-26-high cells, respectively, as target cells. The levels of IFN-γ, IL-2, and TNF-α were determined by ELISA. RESULTS: THL suppressed the growth of CT-26-high tumor in mice previously vaccinated with low-tumorigenic CT-26-low cells or γ-irradiated CT-26-high cells. THL increased the populations of NK cells and CD4+ T lymphocytes in the spleen and enhanced the tumor-killing activities of NK cells and CTL in mice vaccinated with γ-irradiated CT-26-high cells. THL increased the production of IFN-γ, IL-2, and TNF-α in mice vaccinated with γ-irradiated CT-26-high cells. CONCLUSION: THL can enhance the antitumor immune responses in mice vaccinated with killed tumor cells. These results suggest that THL may be used as a complementary medicine for cancer patients previously treated with killed tumor cell vaccines, radiotherapy, or chemotherapy.

Fermentation product of soybean, black bean, and green bean mixture induces apoptosis in a wide variety of cancer cells.

Previous studies have shown that soybean fermentation products can act as cancer chemoprevention or therapeutic agents. In this study, the anticancer activities of a fermentation product of soybean, black bean, and green bean mixture (BN999) were investigated. We found that BN999 inhibited the growth of human breast cancer AU565 cells and prostate adenocarcinoma PC-3 cells but not that of normal human cells. BN999 induced apoptosis in various human cancer cells but not in normal human cells. BN999 treatment of AU565 cancer cells resulted in activation of calpain and caspase-8, -9, and -3, suggesting that BN999 induces apoptosis via receptor-, mitochondria-, and endoplasmic reticulum-mediated pathways. Finally, we showed that BN999 inhibited the growth of mouse CT-26 colon cancer xenografts in syngenic BALB/c mice without causing obvious side effects. Together, these data suggest that BN999 has potential to be used as a cancer chemoprevention or therapeutic agent.

Cellular protein HAX1 interacts with the influenza A virus PA polymerase subunit and impedes its nuclear translocation.

Transcription and replication of the influenza A virus RNA genome occur in the nucleus through the viral RNA-dependent RNA polymerase consisting of PB1, PB2, and PA. Cellular factors that associate with the viral polymerase complex play important roles in these processes. To look for cellular factors that could associate with influenza A virus PA protein, we have carried out a yeast two-hybrid screen using a HeLa cell cDNA library. We identified six cellular proteins that may interact with PA. We focused our study on one of the new PA-interacting proteins, HAX1, a protein with antiapoptotic function. By using glutathione S-transferase pulldown and coimmunoprecipitation assays, we demonstrate that HAX1 specifically interacts with PA in vitro and in vivo and that HAX1 interacts with the nuclear localization signal domain of PA. Nuclear accumulation of PA was increased in HAX1-knockdown cells, and this phenotype could be reversed by reexpression of HAX1, indicating that HAX1 can impede nuclear transport of PA. As a consequence, knockdown of HAX1 resulted in a significant increase in virus yield and polymerase activity in a minigenome assay, and this phenotype could be reversed by reexpression of HAX1, indicating that HAX1 can inhibit influenza A virus propagation. Together, these results not only provide insight into the mechanism underlying nuclear transport of PA but also identify an intrinsic host factor that restricts influenza A virus infection.

SV40 T/t-common polypeptide enhances the sensitivity of HER2-overexpressing human cancer cells to anticancer drugs cisplatin and doxorubicin.

HER2-overexpressing cancer cells are resistant to cisplatin (CDDP) and doxorubicin (DXR). Here we report that SV40 T/t-common polypeptide could specifically sensitize HER2-overexpressing cancer cells to CDDP and DXR and specifically enhance CDDP- or DXR-induced apoptosis in these cells. This activity of T/t-common may be attributed to its ability to inhibit Bcl-2 and Bcl-XL and to suppress ERK activity in CDDP- or DXR-treated HER2-overexpressing cancer cells. T/t-common could enhance the antitumor activity of DXR on HER2-overexpressing ovarian tumor in NOD/SCID mice, suggesting that combination therapy using T/t-common and chemotherapeutic agents may provide a new approach for treating HER2-overexpressing cancers.

Inhibition of metastasis, angiogenesis, and tumor growth by Chinese herbal cocktail Tien-Hsien Liquid.

BACKGROUND: Advanced cancer is a multifactorial disease that demands treatments targeting multiple cellular pathways. Chinese herbal cocktail which contains various phytochemicals may target multiple dys-regulated pathways in cancer cells and thus may provide an alternative/complementary way to treat cancers. Previously we reported that the Chinese herbal cocktail Tien-Hsien Liguid (THL) can specifically induce apoptosis in various cancer cells and have immuno-modulating activity. In this study, we further evaluated the anti-metastatic, anti-angiogenic and anti-tumor activities of THL with a series of in vitro and in vivo experiments. METHODS: The migration and invasion of cancer cells and endothelial cells was determined by Boyden chamber transwell assays. The effect of THL on pulmonary metastasis was done by injecting CT-26 colon cancer cells intravenously to syngenic mice. The in vitro and in vivo microvessel formation was determined by the tube formation assay and the Matrigel plug assay, respectively. The in vivo anti-tumor effect of THL was determined by a human MDA-MB-231 breast cancer xenograft model. The expression of metalloproteinase (MMP)-2, MMP-9, and urokinase plasminogen activator (uPA) was measured by gelatin zymography. The expression of HIF-1alpha and the phosphorylation of ERK1/2 were determined by Western blot. RESULTS: THL inhibited the migration and invasion ability of various cancer cells in vitro, decreased the secretion of MMP-2, MMP-9, and uPA and the activity of ERK1/2 in cancer cells, and suppressed pulmonary metastasis of CT-26 cancer cells in syngenic mice. Moreover, THL inhibited the migration, invasion, and tube formation of endothelial cells in vitro, decreased the secretion of MMP-2 and uPA in endothelial cells, and suppressed neovascularization in Matrigel plugs in mice. Besides its inhibitory effect on endothelial cells, THL inhibited hypoxia-induced HIF-1alpha and vascular endothelial growth factor-A expression in cancer cells. Finally, our results show that THL inhibited the growth of human MDA-MB-231 breast cancer xenografts in NOD-SCID mice. This suppression of tumor growth was associated with decreased microvessel formation and increased apoptosis caused by THL. CONCLUSION: Our data demonstrate that THL had broad-spectra anti-cancer activities and merits further evaluation for its use in cancer therapy.

SV40 T/t-common polypeptide specifically induces apoptosis in human cancer cells that overexpress HER2/neu.

Previously, we reported that SV40 T/t-common polypeptide, which contains the NH(2)-terminal common domain of SV40 large T and small t antigens, can repress HER2/neu (also known as erbB-2) expression and consequently suppress the tumorigenic potential of the HER2/neu-overexpressing ovarian carcinoma cells. Here we report that T/t-common could specifically induce apoptosis in HER2/neu-overexpressing human cancer cell lines but not in nontransformed cell lines and HER2/neu low-expressing human cancer cell lines. The ability of T/t-common to induce apoptosis in HER2/neu-overexpressing cancer cells was derived from its ability to inhibit HER2/neu because reexpression of a large amount of HER2/neu could block apoptosis induced by T/t-common. T/t-common expression in HER2/neu-overexpressing SK-OV-3 cancer cells led to down-regulation of Bcl-2 and Bcl-X(L), and overexpression of Bcl-2 could inhibit the ability of T/t-common to induce apoptosis in these cells. Therefore, the apoptosis-inducing activity of T/t-common is related to its ability to inhibit Bcl-2 expression in HER2/neu-overexpressing cancer cells. Consistent with the apoptosis-inducing activity of T/t-common, we found that T/t-common could specifically inhibit the soft-agarose colony-forming ability of the HER2/neu-overexpressing human cancer cell lines but not that of the HER2/neu low-expressing human cancer cell lines. Finally, we showed that T/t-common could specifically sensitize HER2/neu-overexpressing human cancer cell lines, but not HER2/neu low-expressing human cancer cell lines, to chemotherapeutic agent etoposide. Together, these data suggest that T/t-common alone or in combination with chemotherapy may provide a new approach for treatment of cancers that overexpress HER2/neu.