RESUMEN
Circulating tumor cells (CTCs) play a key role in cancer metastasis but are very difficult to detect. in vivo monitoring CTCs has been recognized as an important technique for cancer research and clinical diagnosis. Recently, a noninvasive method, in vivo flow cytometry (IVFC) has been developed to enable continuous, real-time, and long-duration detection of CTCs in animal models by detecting CTC fluorescence in blood vessels excited by lasers. In this study, we present a simple optical scheme for direct noninvasive CTC detection using confocal microscopes. We demonstrate that line scanning of confocal microscopy can provide effective and quantitative CTC detection in live mice during cancer development. Rare CTC signals can be acquired at the early stage of the tumor development after implantation of subcutaneous tumor and monitored continuously to the end. Signals from CTC clusters can also be acquired and distinguished from single CTCs. Our results suggest confocal microscopy is a simple and reliable method for biologists and doctors to use for cancer research. © 2018 International Society for Advancement of Cytometry.
Asunto(s)
Vasos Sanguíneos/diagnóstico por imagen , Microscopía Confocal/métodos , Células Neoplásicas Circulantes/química , Animales , Biomarcadores de Tumor/sangre , Proteínas Fluorescentes Verdes/química , Humanos , Inyecciones Subcutáneas , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Trasplante de Neoplasias , Células Neoplásicas Circulantes/metabolismo , Células Neoplásicas Circulantes/patología , Células Neoplásicas Circulantes/efectos de la radiación , Células PC-3RESUMEN
Cervical cancer is a major public health concern in China, accounting for almost one-fifth of the global incidence and mortality. The recently prequalified domestic bivalent human papillomavirus (HPV) vaccine offers a practical and feasible preventive measure. In response to the global call for action, the National Health Commission issued an Action Plan to eliminate cervical cancer by 2030, with promotion of the HPV vaccination for school-aged girls as a critical step. Despite this, implementation of the vaccination has been patchy, with very low coverage among eligible girls. To address this, from December 2021 to December 2022, a demonstration project was launched in Shenzhen, Guangdong Province, to promote the inclusion of HPV vaccine in local immunisation programme and to address existing barriers to implementation. Using multiple sources of data, this article presents a case study of the demonstration project, analysing its impact on rolling out HPV vaccination among eligible girls and identifying any challenges encountered during implementation. The demonstration project has shown promising results in increasing the HPV vaccination rate, promoting public awareness and acceptance of the domestic HPV vaccine, and establishing a model for quickly scaling up the vaccination at the municipal level. The success of the project can be attributed to several factors, including strong governmental commitment, sufficient funding, multi-sectoral collaboration, ensured vaccine accessibility and affordability, improved vaccination services, and effective health education and communication strategies. Lessons learned from Shenzhen can provide valuable insights for future advocacy and implementation of the vaccination in other areas of China, but challenges must be addressed to achieve universal coverage. These include addressing vaccine hesitancy, expanding the programme to cover a broader age range, and ensuring consistent quality of vaccination services in primary care facilities. Overcoming these challenges will require innovative strategies, public-private partnerships, and sustained funding and resources. Future research should focus on evaluating the long-term effectiveness of the vaccination programme and identifying contextual factors that may impact its implementation in different settings. Overall, the effective control of cervical cancer in China will rely on the "political will" to ensure the incorporation of preventive interventions into policies and universal programme coverage.
Asunto(s)
Infecciones por Papillomavirus , Vacunas contra Papillomavirus , Neoplasias del Cuello Uterino , Femenino , Humanos , Adolescente , Niño , Anciano , Neoplasias del Cuello Uterino/prevención & control , Infecciones por Papillomavirus/prevención & control , Vacunación , ChinaRESUMEN
The technology to induce stem cell differentiation is of great importance in life science, stem cell therapy, and regenerative medicine. Here, we detail steps to noninvasively activate stem cell differentiation in vitro and in vivo using a tightly focused femtosecond laser. We describe how a single-time transient photoactivation can initiate differentiation without any gene engineering, exogenous substances, or physical contact. This protocol enables the differentiation of adipose-derived stem cells to osteoblasts in vitro and cerebellar granule neuron progenitors to granule neurons in vivo. For complete details on the use and execution of this protocol, please refer to Tang et al. (2022).
Asunto(s)
Osteoblastos , Células Madre , Tejido Adiposo , Diferenciación Celular/genética , Rayos LáserRESUMEN
Tumor cells present anti-apoptosis and abnormal proliferation during development. Senescence and stemness of tumor cells play key roles in tumor development and malignancy. In this study, we show the transient stimulation by a single-time scanning of tightly focused femtosecond laser to tumor cells can modulate the stemness and senescence in vitro and in vivo. The laser-induced cellular senescence and stemness present distinct transitions in vitro and in vivo. The cells 1.2 mm deep in tumor tissue are found with significant senescence induced by the transient photostimulations in 100-200 µm shallow layer in vivo, which suppresses the growth of whole tumor in living mice.
RESUMEN
A variety of physical and chemical methods have been developed in research laboratories for the induction of stem cell differentiation. However, the use of exogenous chemicals and materials may limit their widespread utility in clinics. To develop a clean and precise induction approach with minimal invasion, we reported here that 1-second stimulation by a tightly focused femtosecond laser (fsL) (140 mW/µm2 , 200 fs) can modulate the signaling systems in human mesenchymal cells, such as intracellular calcium and reactive oxygen species. Upon stimulation on an automatic platform, hMSCs were found to express osteoblastic markers and form calcium-rich deposits. Moreover, tissue mineralization was observed when the fsL-illuminated hMSCs were ectopically transplanted into nude mice. Collectively, we described a novel and non-contact optical stimulation method for cell differentiation with high spatiotemporal resolution.
Asunto(s)
Células Madre Mesenquimatosas , Osteogénesis , Animales , Ratones , Humanos , Osteogénesis/fisiología , Calcio , Ratones Desnudos , Diferenciación Celular , Rayos Láser , Células CultivadasRESUMEN
Recent technological advancements on stem cell differentiation induction have been making great progress in stem cell research, regenerative medicine, and therapeutic applications. However, the risk of off-target differentiation limits the wide application of stem cell therapy strategies. Here, we report a non-invasive all-optical strategy to induce stem cell differentiation in vitro and in vivo that activates individual target stem cells in situ by delivering a transient 100-ms irradiation of a tightly focused femtosecond laser to a submicron cytoplasmic region of primary adipose-derived stem cells (ADSCs). The ADSCs differentiate to osteoblasts with stable lineage commitment that cannot further transdifferentiate because of simultaneous initiation of multiple signaling pathways through specific Ca2+ kinetic patterns. This method can work in vivo to direct mouse cerebellar granule neuron progenitors to granule neurons in intact mouse cerebellums through the skull. Hence, this optical method without any genetic manipulations or exogenous biomaterials holds promising potential in biomedical research and cell-based therapies.
Asunto(s)
Tejido Adiposo , Células Madre , Adipocitos/metabolismo , Tejido Adiposo/metabolismo , Animales , Diferenciación Celular/fisiología , Rayos Láser , Ratones , Células Madre/metabolismoRESUMEN
Background: Pulmonary vascular alteration is an important feature of chronic obstructive pulmonary disease (COPD), which is characterized by distal pulmonary vascular pruning in angiography. We aimed to further investigate the clinical relevance of pulmonary vasculature in COPD patients using non-contrast computed tomography (CT). Methods: Seventy-one control subjects and 216 COPD patients completed the questionnaires, spirometry, and computed tomography (CT) scans within 1 month and were included in the study. Small pulmonary vessels represented by percentage of cross-sectional area of pulmonary vessels smaller than 5 mm2 or 5-10 mm2 to the total lung fields (%CSA<5 or %CSA5-10, respectively) were measured using ImageJ software. Spearman correlation was used to investigate the relationship between %CSA<5 and airflow limitation. A receiver operating characteristic (ROC) curve was built to evaluate the value of %CSA<5 in discriminating COPD patients from healthy control subjects. Segmented regression was used to analyze the relationship between %CSA<5 and %LAA-950 (percentage of low-attenuation areas less than -950 HU). Results: We found a significant correlation between %CSA<5 and forced expiratory volume in one second (FEV1) percentage of predicted value (%pred) (r = 0.564, P < 0.001). The area under the ROC curve for the value of %CSA<5 in distinguishing COPD was 0.816, with a cut-off value of 0.537 (Youden index J, 0.501; sensitivity, 78.24%; specificity, 71.83%). Since the relationship between %CSA<5 and %LAA-950 was not constant, performance of segmented regression was better than ordinary linear regression (adjusted R2, 0.474 vs 0.332, P < 0.001 and P < 0.001, respectively). As %CSA<5 decreased, %LAA-950 slightly increased until an inflection point (%CSA<5 = 0.524) was reached, after which the %LAA-950 increased apparently with a decrease in %CSA<5. Conclusion: %CSA<5 was significantly correlated with both airflow limitation and emphysema, and we identified an inflection point for the relationship between %CSA<5 and %LAA-950.
Asunto(s)
Enfisema , Enfermedad Pulmonar Obstructiva Crónica , Enfisema Pulmonar , Volumen Espiratorio Forzado , Humanos , Pulmón , Enfermedad Pulmonar Obstructiva Crónica/diagnóstico por imagen , Enfisema Pulmonar/diagnóstico por imagen , Enfisema Pulmonar/etiología , Tomografía Computarizada por Rayos X/métodos , Capacidad VitalRESUMEN
Dysregulation of the PINK1/Parkin-mediated mitophagy is essential to Parkinson's disease. Although important progress has been made in previous researches, the biochemical reagents that induce global and significant mitochondrial damage may still hinder deeper insights into the mechanisms of mitophagy. The origin of PINK1/Parkin pathway activation in mitophagy remains elusive. In this study, we develop an optical method, ultra-precise laser stimulation (UPLaS) that delivers a precise and noninvasive stimulation onto a submicron region in a single mitochondrial tubular structure. UPLaS excites localized mitochondrial Ca2+ (mitoCa2+) oscillations with tiny perturbation to mitochondrial membrane potential (MMP) or mitochondrial reactive oxygen species. The UPLaS-induced mitoCa2+ oscillations can directly induce PINK1 accumulation and Parkin recruitment on mitochondria. The Parkin recruitment by UPLaS requires PINK1. Our results provide a precise and noninvasive technology for research on mitophagy, which stimulates target mitochondria with little damage, and reveal mitoCa2+ oscillation directly initiates the PINK1-Parkin pathway for mitophagy without MMP depolarization.
Asunto(s)
Señalización del Calcio , Calcio/metabolismo , Mitocondrias/enzimología , Mitofagia , Proteínas Quinasas/metabolismo , Ubiquitina-Proteína Ligasas/metabolismo , Células HeLa , Humanos , Potencial de la Membrana Mitocondrial , Microscopía Confocal , Microscopía Fluorescente , Mitocondrias/patología , Proteínas Quinasas/genética , Especies Reactivas de Oxígeno , Análisis de la Célula Individual , Factores de Tiempo , Imagen de Lapso de Tiempo , Ubiquitina-Proteína Ligasas/genéticaRESUMEN
Ca2+ channels are essential to cell birth, life, and death. They can be externally activated by optogenetic tools, but this requires robust introduction of exogenous optogenetic genes for expression of photosensitive proteins in biological systems. Here we present femtoSOC, a method for direct control of Ca2+ channels solely by ultrafast laser without the need for optogenetic tools or any other exogenous reagents. Specifically, by focusing and scanning wavelength-tuned low-power femtosecond laser pulses on the plasma membrane for multiphoton excitation, we directly induced Ca2+ influx in cultured cells. Mechanistic study reveals that photoexcited flavins covalently bind cysteine residues in Orai1 via thioether bonds, which facilitates Orai1 polymerization to form store-operated calcium channels (SOCs) independently of STIM1, a protein generally participating in SOC formation, enabling all-optical activation of Ca2+ influx and downstream signaling pathways. Moreover, we used femtoSOC to demonstrate direct neural activation both in brain slices in vitro and in intact brains of living mice in vivo in a spatiotemporal-specific manner, indicating potential utility of femtoSOC.