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1.
Gynecol Endocrinol ; 40(1): 2317270, 2024 Feb 12.
Artículo en Inglés | MEDLINE | ID: mdl-38518807

RESUMEN

AIMS: The aim of this study was to investigate the impact of three single nucleotide polymorphisms (SNPs) within X-Ray Repair Cross Complementary Group 2 (XRCC2) gene and additional gene- abdominal obesity (AO) interaction with endometrial carcinoma (EC) risk. METHODS: Hardy-Weinberg equilibrium was tested for all participants by using SNPstats (online software: http://bioinfo.iconcologia.net/SNPstats). The best SNP-SNP and gene-AO interaction combination among three SNPs within XRCC2 gene and AO was screened using generalized multifactor dimensionality reduction (GMDR). RESULTS: We employed the logistic regression analysis showed that rs718282-T allele is associated with increased EC risk, adjusted ORs (95%CI) were 1.67 (1.23-2.04). However, we did not find statistical association between rs3218536, and rs3218384 and EC susceptibility. GMDR analysis was used for SNP-SNP- and gene-abdominal obesity analysis. The cross-validation consistency and the testing accuracy for the interaction were calculated. The two-locus model between rs718282 and AO had a testing accuracy of 60.11%, which was significant at the p < .001 level, and this two- locus model was considered as the best model. It provided statistical evidence for rs718282 gene-AO interaction effects. The results indicated that AO influenced the EC risk depending on the rs718282 genotypes. Compared with non- AO subjects with rs718282-CC genotype, AO subjects with rs718282-CT or TT genotype had the highest EC risk, OR (95%CI) was 2.83 (1.67 - 4.02), after covariates adjustment. CONCLUSIONS: Both the rs718282- T allele, and its interaction with AO were associated with increased EC risk.


Asunto(s)
Neoplasias Endometriales , Predisposición Genética a la Enfermedad , Humanos , Femenino , Obesidad Abdominal/complicaciones , Obesidad Abdominal/epidemiología , Obesidad Abdominal/genética , Rayos X , Genotipo , Obesidad/complicaciones , Obesidad/epidemiología , Obesidad/genética , Polimorfismo de Nucleótido Simple , Neoplasias Endometriales/epidemiología , Neoplasias Endometriales/genética , China , Estudios de Casos y Controles , Proteínas de Unión al ADN/genética
2.
Artículo en Inglés | MEDLINE | ID: mdl-38743892

RESUMEN

Objective: To improve the preoperative management of patients undergoing day arthroscopy. Methods: Based on the evidence-based concept, the preoperative management strategy of daytime arthroscopic surgery was practiced, and the implementation effect of patients undergoing daytime arthroscopic surgery before and after evidence application was compared. The evidence-based strategies adopted in this study include: informed consent,seven aspects: anesthesia assessment, health education, prohibition of drinking and eating, preoperative medication guidance, surgical arrangement, and emergency management. Results: After applying evidence-based strategies, significant improvements were observed in various quality indicators, including reduced waiting time, reduced surgical cancellations, increased patient satisfaction, and reduced unplanned hospitalization rates. Conclusion: Based on the evidence-based concept, the practice of preoperative management strategies for patients undergoing daytime arthroscopic surgery optimizes the daytime surgical process from the strict implementation of patient health education and evaluation, as well as the establishment of compliant and reasonable surgical schedules and emergency mechanisms. This improves the preoperative management behavior of daytime surgery, enhances the patient's medical experience, and enhances the quality of daytime surgical care, providing a good basis for management and decision-making.

3.
Zhongguo Zhong Yao Za Zhi ; 49(3): 735-743, 2024 Feb.
Artículo en Zh | MEDLINE | ID: mdl-38621877

RESUMEN

Chemical constituents of 70% ethanol extract of Alangium chinense subsp. pauciflorum were investigated. The 70% ethanol extract of A. chinense subsp. pauciflorum was isolated and purified by D-101 macroporous resins, silica gel, Sephadex LH-20 and other methods. As a result, nineteen compounds were isolated and identified as 4-cyclohexene-1α,2α,3α-triol-1-O-ß-D-glucoside(1), 1ß,4α,6α,13-tetrahydroxy-eudesm-11(12)-ene(2), sucrose(3), 1'-O-benzyl-α-L-rhamnopyranosyl-(1″→6')-ß-D-glucopyranoside(4), bis(2-ethylhexyl)benzene-1,2-dicarboxylate(5),(Z)-10-heneicosenoic acid(6), di-O-methylcrenati(7), methyl-α-D-fructofuranoside(8), ß-daucosterol(9), syringic acid(10), vanillicacid(11), octacosanol(12), isoarborinol(13), 2,7-dihydroxy-6-methyl-4-(1-methylethyl)-1-naphthalenecarboxylate(14),vanillin(15), coniferyl aldehyde(16), 9(11)-dehydroergosterolperoxide(17), 5α,8α-epidioxy-(22E,24R)-ergosta-6,22-dien-3ß-ol(18), ß-sitosterol(19), respectively. Compounds 1 and 2 were new compounds, compounds 5-11, 13, 15-18 were isolated from Alangium for the first time.The anti-inflammatory activity of compourd 1 was determinded by the LPS-induced RAW264.7 macrophage inflammation model. The results showed that the new compound 1 has a certain inhibitory effect on LPS-induced NO production of RAW264.7 cells, and the inhibitory rate was 54.57%.


Asunto(s)
Alangiaceae , Lipopolisacáridos , Antiinflamatorios/farmacología , Etanol , Extractos Vegetales
4.
Funct Integr Genomics ; 23(1): 39, 2023 Jan 16.
Artículo en Inglés | MEDLINE | ID: mdl-36642729

RESUMEN

Ovarian cancer (OC) is the most common malignant cancer in the female reproductive system. Hypoxia is an important part of tumor immune microenvironment (TIME), which is closely related to cancer progression and could significantly affect cancer metastasis and prognosis. However, the relationship between hypoxia and OC remained unclear. OCs were molecularly subtyped by consensus clustering analysis based on the expression characteristics of hypoxia-related genes. Kaplan-Meier (KM) survival was used to determine survival characteristics across subtypes. Immune infiltration analysis was performed by using Estimation of Stromal and Immune cells in Malignant Tumors using Expression data (ESTIMATE) and microenvironment cell populations-counter (MCP-Counter). Differential expression analysis was performed by using limma package. Next, univariate Cox and least absolute shrinkage and selection operator (LASSO) regression analyses were used to build a hypoxia-related risk score model (HYRS). Mutational analysis was applied to determine genomic variation across the HYRS groups. The Tumor Immune Dysfunction and Exclusion (TIDE) algorithm was used to compare the effectiveness of HYRS in immunotherapy prediction. We divided OC samples into two molecular subtypes (C1 and C2 subtypes) based on the expression signature of hypoxia genes. Compared with C1 subtype, there was a larger proportion of poor prognosis genotypes in the C2 subtype. And most immune cells scored higher in the C2 subtype. Next, we obtained a HYRS based on 7 genes. High HYRS group had a higher gene mutation rate, such as TP53. Moreover, HYRS performed better than TIDE in predicting immunotherapy effect. Combined with clinicopathological features, the nomogram showed that HYRS had the greatest impact on survival prediction and a strong robustness.


Asunto(s)
Neoplasias Ováricas , Femenino , Humanos , Neoplasias Ováricas/genética , Hipoxia/genética , Algoritmos , Análisis por Conglomerados , Genotipo , Microambiente Tumoral/genética
5.
Anal Chem ; 95(2): 889-897, 2023 01 17.
Artículo en Inglés | MEDLINE | ID: mdl-36537841

RESUMEN

The time since deposition (TSD) of latent fingermarks (LFMs) serves as "witnesses" for crime scene reconstructions. Nevertheless, existing TSD prediction approaches focused on either physical or chemical aging parameters leading to inaccurate estimation. A novel label-free protocol has been developed, where both physical ridge patterns and lipid oxide (LipOx) degradation kinetics are realized using optical microscopy and scanning electrochemical microscopy (SECM) and combined for TSD prediction. Specifically, the surface interrogation (SI)-SECM titration was utilized to monitor the LipOx degradation in LFM arrays aligned by hole array masks, through which we derived the LipOx degradation function. After establishing the relationship between several titration parameters and titrated area by experimental and numerical simulation methods, the titrated area could be reasonably estimated and subsequently used to calculate the surface coverage of LipOx. Results demonstrated that the tip transient revealed the LipOx coverage of deposited LFMs. Notably, LipOx coverage was found to increase during the first day and then decrease over time, whose degradation rate was susceptible to light. Thus, TSD candidates of an LFM could be limited to two values through the established function. Due to the nonmonotonic trend of LipOx aging, a physical parameter "the gray value ratio (GVR) of furrows to ridges" was proposed to exclude irrelevant TSD through support vector machine (SVM) classification. Ultimately, we predicted TSDs of seven LFMs with estimation errors of 2.2-26.8%. Overall, our strategy, with the outperformed capability of gleaning physical and electrochemical information on LFMs, can provide a truly label-free way of studying LFMs and hold great promise for multidimensional fingerprint information analysis.


Asunto(s)
Bezafibrato , Microscopía
6.
Anal Chem ; 95(50): 18470-18478, 2023 12 19.
Artículo en Inglés | MEDLINE | ID: mdl-38051701

RESUMEN

Blood is one of the most frequent and valuable traces encountered at crime scenes, where knowing the time since deposition (TSD) of bloodstains tremendously assists forensic experts to screen out crime-related evidence and aids in the reconstruction of the event sequence. Although increasing proof-of-concept methodologies for investigating the TSD of bloodstains have been reported, there is still no accepted strategy in forensic practice as the aging mechanism involves complex components, leading to the inaccuracy of the estimation results. Herein, an endogenous biomarker of alkaline phosphatase (ALP) was chosen to investigate the TSD by scanning electrochemical microscopy (SECM). Results demonstrate that the ALP activity acquired via SECM lateral scan assay exhibited a clear decrease over time, and a similar trend was observed on both poly(vinylidene fluoride) (PVDF) membrane and glass, with the aging kinetics on PVDF membrane being faster than glass. By means of quantitatively calculating the flux of generated p-aminophenol (PAP), we established the aging curve and realized the TSD estimation of blood fingerprints (BFPs) that was unable to be distinguished via optical measurements. Intriguingly, the as-obtained estimation accuracy ranged from 74.6 to 93.7%, proving the possibility of using an ALP biomarker and SECM. More appealingly, the predicted TSDs were capable of accurately differentiating the deposition sequence of overlapping BFPs, which was hardly achieved by optical means. Therefore, this proof-of-concept strategy demonstrates the value of SECM as a forensic tool and opens possibilities for revealing multidimensional information about crime.


Asunto(s)
Fosfatasa Alcalina , Polivinilos , Microscopía Electroquímica de Rastreo , Biomarcadores
7.
Anal Chem ; 95(39): 14686-14694, 2023 Oct 03.
Artículo en Inglés | MEDLINE | ID: mdl-37713524

RESUMEN

The problem of corrosion-induced discoloration and embrittlement in silverware is a significant concern for the long-term preservation of excavated archeological silver artifacts, even after thermal restoration. The key to addressing this issue lies in the meticulous selection and evaluation of corrosion inhibitors that possess targeted corrosion inhibition capabilities. This study focuses on the evaluation of corrosion inhibitors for archeological silver artifacts using scanning electrochemical cell microscopy (SECCM) and X-ray photoelectron spectroscopy (XPS). The researchers aimed to compare the inhibition effects of four corrosion inhibitors [1,2,3-benzotriazole (BTA), 2-mercaptobenzimidazole (MBI), 2-mercaptobenzothiazole (MBT), and 2-mercaptobenzoxazole (MBO)] on a simulated Ag-Cu alloy sample and understand their mechanisms. The results showed that MBT exhibited better corrosion inhibition for microstructural regions with higher silver content due to its ability to form stable chelation structures with Ag(I). MBO exhibited better corrosion inhibition for microstructural regions with higher copper content due to its strong affinity with Cu(I). The targeted corrosion inhibition ability for the ß-phase was ranked as MBO > BTA ≈ MBI > MBT, while for the α-phase the ranking was MBT > MBO > MBI > BTA. The study demonstrated the feasibility and capabilities of SECCM in the targeted screening of corrosion inhibitors for different compositions and microstructural regions in archeological metal artifacts. This study highlights the potential of SECCM in corrosion inhibitor research for archeological metal artifacts and wider applications in metal material corrosion protection.

8.
Analyst ; 148(5): 1032-1040, 2023 Feb 27.
Artículo en Inglés | MEDLINE | ID: mdl-36723182

RESUMEN

Fingerprints have long been the gold standard for personal identification in forensic science. However, realizing the high-resolution enhancement of eccrine LFPs is difficult using the traditional methods and the label-free detection of fingerprint residue information is also challenging. Herein, we propose two enhancement strategies for LFPs on PVDF membrane (LFPs/PVDF) using blue-black ink staining and scanning electrochemical microscopy (SECM). The blue-black ink staining method was used for the first time to develop three types (sebaceous, natural and eccrine) of LFPs/PVDF based on the difference in wettability between the fingerprint residues and PVDF membrane. The enhanced fingerprints clearly displayed levels 1-3 features with high contrast and low background interference. Furthermore, we achieved chemical imaging of the LFP/PVDF samples, where their possible visualization mechanisms were ascribed to the electrochemical reactivity of squalene and difference in wettability between the LFP and PVDF membrane, which was first proposed and investigated by SECM imaging and water contact angle (WCA) measurements, respectively. Significantly, SECM imaging not only provided fingerprint patterns without any labelling but also revealed the spatial distribution information of squalene in LFPs simultaneously. In addition, it was also demonstrated that LFPs deposited on various surfaces were first successfully transferred to the PVDF membrane, and then further developed with both methods, making them general for personal identity-related applications. Taken together, the blue-black ink staining method can easily and quickly obtain level 3 features of LFPs/PVDF and the SECM approach can non-invasively image the topography and chemical information of LFPs/PVDF, and thus they can be potentially selected according to various requirements in forensic scenarios.

9.
Analyst ; 148(11): 2438-2448, 2023 May 30.
Artículo en Inglés | MEDLINE | ID: mdl-37139711

RESUMEN

A facile and high-resolution enhancement of latent fingerprints (LFPs) has been developed by using a wet nitrocellulose (NC) membrane as a matrix under natural light. A clear fingerprint pattern was presented on the membrane after a fingertip touch owing to the difference in light transmittance between the ridge residues and the wet-NC-membrane background. Compared with conventional methods, this protocol can provide a higher resolution fingerprint image to extract level 3 details accurately. It is also compatible with commonly used fingerprint visualization techniques (magnetic ferric oxide powder and AgNO3. The modified membrane could be more general to realize the high-resolution visualization of LFP transferred from various substrates, even independent of light projection. Due to the excellent feasibility and reproducibility of level 3 details extracted by the wet NC membrane, the frequency distribution of the distance between adjacent sweat pores (FDDasp) could be used to effectively distinguish the fragmentary fingerprints. Finally, the level 3 features of LFPs from females and males were conveniently extracted by the wet-NC-membrane method for gender identification. The statistical results indicated that females had a higher average sweat pore density (115/9 mm2) than males (84/9 mm2). Taken together, this approach provided a high-resolution, reproducible, and accurate imaging of LFPs, which shows great promise for forensic information analysis.


Asunto(s)
Dermatoglifia , Masculino , Humanos , Colodión , Reproducibilidad de los Resultados
10.
J Cell Mol Med ; 26(15): 4292-4304, 2022 08.
Artículo en Inglés | MEDLINE | ID: mdl-35789100

RESUMEN

Nonsyndromic cleft palate only (NSCP) is a common congenital malformation worldwide. In this study, we report a three-generation pedigree with NSCP following the autosomal-dominant pattern. Whole-exome sequencing and Sanger sequencing revealed that only the frameshift variant c.1012dupG [p. E338Gfs*26] in PARD3 cosegregated with the disease. In zebrafish embryos, ethmoid plate patterning defects were observed with PARD3 ortholog disruption or expression of patient-derived N-terminal truncating PARD3 (c.1012dupG), which implicated PARD3 in ethmoid plate morphogenesis. PARD3 plays vital roles in determining cellular polarity. Compared with the apical distribution of wild-type PARD3, PARD3-p. E338Gfs*26 mainly localized to the basal membrane in 3D-cultured MCF-10A epithelial cells. The interaction between PARD3-p. E338Gfs*26 and endogenous PARD3 was identified by LC-MS/MS and validated by co-IP. Immunofluorescence analysis showed that PARD3-p. E338Gfs*26 substantially altered the localization of endogenous PARD3 to the basement membrane in 3D-cultured MCF-10A cells. Furthermore, seven variants, including one nonsense variant and six missense variants, were identified in the coding region of PARD3 in sporadic cases with NSCP. Subsequent analysis showed that PARD3-p. R133*, like the insertion variant of c.1012dupG, also changed the localization of endogenous full-length PARD3 and that its expression induced abnormal ethmoid plate morphogenesis in zebrafish. Based on these data, we reveal PARD3 gene variation as a novel candidate cause of nonsyndromic cleft palate only.


Asunto(s)
Labio Leporino , Fisura del Paladar , Animales , Cromatografía Liquida , Labio Leporino/genética , Fisura del Paladar/genética , Predisposición Genética a la Enfermedad , Polimorfismo de Nucleótido Simple , Espectrometría de Masas en Tándem , Pez Cebra/genética
11.
Anal Chem ; 94(49): 17320-17327, 2022 12 13.
Artículo en Inglés | MEDLINE | ID: mdl-36448925

RESUMEN

A simple and universal strategy for fabricating flexible 25 µm platinum (Pt) disk ultramicroelectrodes (UMEs) was proposed, where a pulled borosilicate glass micropipette acted as a mold for shaping the flexible tip with flexible epoxy resin. The whole preparation procedure was highly efficient, enabling 10 or more probes to be manually fabricated within 10 h. Intriguingly, this technique permits an adjustable RG ratio, tip length, and stiffness, which could be tuned according to varying experimental demands. Besides, the electroactive area of the probe could be exposed and made renewable with a thin blade, allowing its reuse in multiple experiments. The flexibility characterization was then employed to optimize the resin/hardener mass ratio of epoxy resin and the tip position during HF etching in the fabrication process, suggesting that more hardener, a larger RG value, or a longer tip length obtained stronger deformation resistance. Subsequently, the as-prepared probe was examined by optical microscopy, cyclic voltammetry, and SECM approach curves. The results demonstrated the probe possessed good geometry with a small RG ratio of less than 3 and exceptional electrochemical properties, and its insulating sheath remained undeformed after blade cutting. Owing to the tip's flexibility, it could be operated in contactless mode with an extremely low working distance and even in contact mode scanning to achieve high spatial resolution and high sensitivity while guaranteeing that the tip and samples would suffer minimal damage if the tip crashed. Finally, the flexible probe was successfully employed in three scanning scenarios where tilted and 3D structured PDMS microchips, a latent fingerprint deposited on the stiff copper sheet, and soft egg white were included. In all, the flexible probe encompasses the advantages of traditional disk UMEs and circumvents their principal drawbacks of tip crash and causing sample scratches, which is thus more compatible with large specimens of 3D structured, stiff, or even soft topography.


Asunto(s)
Resinas Epoxi , Platino (Metal) , Microscopía Electroquímica de Rastreo , Electroquímica/métodos , Platino (Metal)/química , Microscopía/métodos
12.
Retina ; 42(9): 1702-1708, 2022 09 01.
Artículo en Inglés | MEDLINE | ID: mdl-35576609

RESUMEN

BACKGROUND: Retinal detachment is a severe complication of acute retinal necrosis, threatening vision. The published articles on the effectiveness of laser in preventing retinal detachment in acute retinal necrosis are controversial. Therefore, we aim to evaluate whether prophylactic laser is effective for retinal detachment after acute retinal necrosis. METHODS: PubMed, Embase, and Cochrane databases were searched, and the retrieved records were screened. Each included study has well-defined laser-treated group and control group without laser treatment or with enough data for manual grouping. The quality of the included studies was assessed using ROBINS-I ("Risk Of Bias In Nonrandomized Studies-of Interventions"). Meta-analysis was conducted to calculate the pooled odds ratios and their 95% confidence interval. Sensitivity analysis was used to test the solidarity, and subgroup analysis was performed to determine the source of heterogeneity. RESULTS: Fourteen studies with a total of 532 eyes were eventually included. The quality of the included studies was moderate. The combined results showed that the pooled odds ratio was 0.61 (95% confidence interval [0.41∼0.90], P < 0.05, I 2 = 27%). Sensitivity analysis showed that the odds ratios were similar when excluding any study. Subgroup analysis showed moderate heterogeneity among three subgroups (I 2 = 48.0%, P = 0.15 for heterogeneity), and the odds ratio in antiviral therapy and steroid combined with prophylactic laser was 0.43 (95% confidence interval [0.25∼0.74], P < 0.05, I 2 = 0%). CONCLUSION: In patients with acute retinal necrosis, laser photocoagulation is an effective treatment to prevent retinal detachment, especially in the subgroup combined with antiviral therapy and steroid.


Asunto(s)
Desprendimiento de Retina , Síndrome de Necrosis Retiniana Aguda , Antivirales , Humanos , Coagulación con Láser/métodos , Desprendimiento de Retina/etiología , Desprendimiento de Retina/prevención & control , Desprendimiento de Retina/cirugía , Síndrome de Necrosis Retiniana Aguda/complicaciones
13.
Anal Chem ; 93(36): 12417-12425, 2021 09 14.
Artículo en Inglés | MEDLINE | ID: mdl-34464090

RESUMEN

The cellular heterogeneity and plasticity are often overlooked due to the averaged bulk assay in conventional methods. Optical imaging-based single-cell analysis usually requires specific labeling of target molecules inside or on the surface of the cell membrane, interfering with the physiological homeostasis of the cell. Scanning electrochemical microscopy (SECM), as an alternative approach, enables label-free imaging of single cells, which still confronts the challenge that the long-time scanning process is not feasible for large-scale analysis at the single-cell level. Herein, we developed a methodology combining a programmable SECM (P-SECM) with an addressable microwell array, which dramatically shortened the time consumption for the topography detection of the micropits array occupied by the polystyrene beads as well as the evaluation of alkaline phosphatase (ALP) activity of the 82 single cells compared with the traditional SECM imaging. This new arithmetic was based on the line scanning approach, enabling analysis of over 900 microwells within 1.2 h, which is 10 times faster than conventional SECM imaging. By implementing this configuration with the dual-mediator-based voltage-switching (VSM) mode, we investigated the activity of ALP, a promising marker for cancer stem cells, in hundreds of tumor and stromal cells on a single microwell device. The results discovered that not only a higher ALP activity is presented in cancer cells but also the heterogeneous distribution of kinetic constant (kf value) of ALP activity can be obtained at the single-cell level. By directly relating large numbers of addressed cells on the scalable microfluidic device to the deterministic routing of the above SECM tip, our platform holds potential as a high-content screening tool for label-free single-cell analysis.


Asunto(s)
Dispositivos Laboratorio en un Chip , Análisis de la Célula Individual , Microscopía Electroquímica de Rastreo , Imagen Óptica
14.
Analyst ; 146(1): 33-47, 2021 Jan 04.
Artículo en Inglés | MEDLINE | ID: mdl-33140753

RESUMEN

Fingermarks have long been recognized as one of the most reliable and valuable evidence for personal identification. In practice, fingerprint analysis primarily concentrates on latent fingerprint visualization. However, fingerprint visualization techniques do not always enable individualization when fingermarks collected in crime scenes are fragmentary, ambiguous, or deformed. Age determination techniques based on physical and chemical composition changes in fingerprints over time have attracted researchers' attention in recent years. Nevertheless, the components of fingerprints are liable to factors including donor features, deposition conditions, substrate properties, environmental conditions and revealing methods. All the influences mainly contribute to unreliable outcomes of age estimation. Recent developments in fingermark age determination have moved forward to more precise approaches. The advanced methods can be classified into two categories including techniques based on the modifications of physical characteristics and chemical composition characteristics. Herein, the review includes the five types of variables that influence the aging process. The methodologies are subsequently highlighted along with their advantages and disadvantages. Furthermore, photography, optical, microscopy and electrochemical methods, and vibrational spectroscopy and mass spectrometry (MS) techniques are summarized in detail, with an emphasis on their utilization.


Asunto(s)
Dermatoglifia , Técnicas Electroquímicas , Espectrometría de Masas
15.
Anal Chem ; 92(11): 7638-7645, 2020 06 02.
Artículo en Inglés | MEDLINE | ID: mdl-32374153

RESUMEN

Three-dimensional cell culture provides an efficient way to simulate the in vivo tumorigenic microenvironment where tumor-stroma interaction intrinsically plays a pivotal role. Conventional three-dimensional (3D) culture is inadequate to address precise coexistential heterogeneous pairing and quantitative measurement in a parallel algorithm format. Herein, we implemented a set of microwell array microfluidic devices to study the cell spheroids-based tumor-stromal metastatic process in vitro. This approach enables accurate one-to-one pairing between tumor and fibroblast spheroid for dissecting 3D tumor invasion in the manner of high-content imaging. On one single device, 240 addressable tumor-stroma pairings can be formed with convenient pipetting and centrifugation within a small area of 1 cm2. Consequential confocal imaging analysis disclosed that the tumor spheroid could envelop the fibroblast spheroid. Specific chemicals can effectively hamper or promote this 3D metastasis. Due to the addressable time-resolved measurements of the merging process of hundreds of doublets, our approach allows us to decipher the metastatic phenotype between different tumor spheroids. Compared with traditional protocols, massive heterogeneous cellular spheroids pairing and merging using this method is well-defined with microfluidic control, which leads to a favorable high-content tumor-stroma doublet metastasis analysis. This simple technique will be a useful tool for investigating heterotypic spheroid-spheroid interactions.


Asunto(s)
Dispositivos Laboratorio en un Chip , Neoplasias/patología , Esferoides Celulares/patología , Células del Estroma/patología , Humanos , Células Tumorales Cultivadas , Microambiente Tumoral
16.
Int Urogynecol J ; 31(1): 15-33, 2020 01.
Artículo en Inglés | MEDLINE | ID: mdl-31485686

RESUMEN

INTRODUCTION AND HYPOTHESIS: Pelvic organ prolapse (POP) is a common medical condition universally. In addition to physical examination, experts have increasingly turned their attention to ultrasound in diagnosing POP for its low cost and dynamic imaging. The aim of this paper is to provide an overview of the methods of pelvic floor ultrasound in diagnosing POP, which has been lacking up till now. METHODS: We included original papers comparing the outcome of the Pelvic Organ Prolapse Quantification system and ultrasound, published from 2008 to present in English, using electronic databases (MEDLINE, EMBASE, CENTRAL, PUBMED). All stages of the review were conducted in parallel by two reviewers. RESULTS: Fifteen papers were included. We found that current methods have advantages and limitations. The main methods are to measure levator hiatus-related parameters and distances between the lowest point of the pelvic organs and reference lines during Valsalva maneuver, contraction, and at rest. CONCLUSIONS: Pelvic floor ultrasound is valuable in diagnosing POP, yet suffers from a weakness in precision compared with physical examination. From the existing research, we found that the differences in baseline data such as weight, height, ethnicity, etc., may affect the cutoffs of the above-mentioned parameters. Further research is required to find one appropriate cutoff for each parameter, even if it is necessary to set group values for every parameter according to varying situations.


Asunto(s)
Diafragma Pélvico/diagnóstico por imagen , Prolapso de Órgano Pélvico/diagnóstico por imagen , Ultrasonografía , Adulto , Anciano , Femenino , Humanos , Persona de Mediana Edad , Examen Físico , Valor Predictivo de las Pruebas , Sensibilidad y Especificidad
17.
J Cell Mol Med ; 23(10): 6978-6988, 2019 10.
Artículo en Inglés | MEDLINE | ID: mdl-31410985

RESUMEN

Recent studies have demonstrated that aberrant long non-coding RNAs (lncRNAs) expression are suggested to be closely associated with multiple human diseases, lung cancer included. However, the roles of lncRNAs in lung cancer are not well understood. In this study, we used microarrays to investigate the aberrantly expressed lncRNAs in the mouse lung adenocarcinoma with P53 knockout and the KrasG12D mutation. Results revealed that 6424 lncRNAs were differentially expressed (≥ 2-fold change, P < .05). Two hundred and ten lncRNAs showed more than 8-fold change and conserved across human and were further analysed in the primary mouse lung adenocarcinoma KP cells, which were isolated from the p53 knockout and the KrasG12D mutation mice. Among all the 210 lncRNAs, 11 lncRNAs' expression was regulated by P53, 33 lncRNAs by KRAS and 13 lncRNAs by hypoxia in the primary KP cells, respectively. NONMMUT015812, which was remarkably up-regulated in the mouse lung adenocarcinoma and negatively regulated by the P53 re-expression, was detected to analyse its cellular function. Results showed that knockdown of NONMMUT015812 by shRNAs decreased proliferation and migration abilities of KP cells. Among those aberrantly expressed lncRNAs in the mouse lung adenocarcinoma, NONMMUT015812 was a potential oncogene.


Asunto(s)
Adenocarcinoma del Pulmón/genética , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Neoplasias Pulmonares/genética , Mutación/genética , Proteínas Proto-Oncogénicas p21(ras)/genética , ARN Largo no Codificante/genética , Proteína p53 Supresora de Tumor/genética , Adenocarcinoma del Pulmón/patología , Animales , Secuencia de Bases , Hipoxia de la Célula/genética , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Células HEK293 , Humanos , Neoplasias Pulmonares/patología , Ratones Noqueados , Oncogenes , ARN Largo no Codificante/metabolismo
18.
Anal Chem ; 91(7): 4307-4311, 2019 04 02.
Artículo en Inglés | MEDLINE | ID: mdl-30869520

RESUMEN

The 3D cell spheroid is an emerging tool that allows better recapitulating of in vivo scenarios with multiple factors such as tissue-like morphology and membrane protein expression that intimately coordinates with enzyme activity, thus providing a psychological environment for tumorigenesis study. For analyzing different spheroids, conventional optical imaging may be hampered by the need for fluorescent labeling, which could cause toxicity side effects. As an alternative approach, scanning electrochemical microscopy (SECM) enables label-free imaging. However, SECM for cell spheroid imaging is currently suffering from incapability of systematically analyzing the cell aggregates from spheroid generation, electrochemical signal gaining, and the gene expression on different individual cell spheroids. Herein, we developed a top-removable microfluidic device for cell aggregate yielding and SECM imaging methodology to analyze heterotypic 3D cell spheroids on a single device. This technique allows not only on-chip culturing of cell aggregates but also SECM imaging of the spheroids after opening the chip and subsequent qPCR assay of corresponding clusters. Through employment of the micropit arrays (85 × 4) with a top withdrawable microfluidic layer, uniformly sized breast tumor cell and fibroblast spheroids can be simultaneously produced on a single device. By leveraging voltage-switching mode SECM at different potentials of dual mediators, we evaluated alkaline phosphatase without disturbance of substrate morphology for distinguishing the tumor aggregates from stroma. Moreover, this method also enables gene expression profiling on individual tumor or stromal spheroids. Therefore, this new strategy can seamlessly bridge SECM measurements and molecular biological analysis.


Asunto(s)
Fosfatasa Alcalina/análisis , Dispositivos Laboratorio en un Chip , Técnicas Analíticas Microfluídicas/métodos , Microscopía Electroquímica de Rastreo/métodos , Esferoides Celulares/química , Fosfatasa Alcalina/genética , Biomarcadores de Tumor/análisis , Biomarcadores de Tumor/genética , Técnicas de Cultivo de Célula/instrumentación , Técnicas de Cultivo de Célula/métodos , Fibroblastos/química , Perfilación de la Expresión Génica , Humanos , Células MCF-7 , Técnicas Analíticas Microfluídicas/instrumentación , Prueba de Estudio Conceptual , ARN Mensajero/análisis , Reacción en Cadena en Tiempo Real de la Polimerasa
19.
Anal Chem ; 90(1): 777-784, 2018 01 02.
Artículo en Inglés | MEDLINE | ID: mdl-29191012

RESUMEN

Cancer metastatic dissemination is a complex event during tumor progression which involves cell-cell and cell-matrix interactions. Micropatterning is one of the most efficient ways to study tumor development because it can tune the distribution of cells with spatial and temporal control. Extensive studies have shown that microfluidics can provide a feasible method for cell patterning. However, the current technique requires a microfabrication laboratory to manufacture the chip, which results in inaccessibility to researchers, especially biologists who focus on disclosing biological mechanisms rather than the methods. In this work, we developed a new methodology (tape-assisted photolithographic-free microfluidic chip, TAPMiC) that can realize homogeneous and heterogeneous micropatterning (45 features, 300 µm diameter of each) on a culture dish without the photolithographic procedure. We have applied this method to study critical biological problems, such as tumor cell migration under different conditions, including antitumor pharmaceutics and candidate gene RNAi assay that was relevant to tumor translocation and invasion. Moreover, this platform can achieve copatterning to recapitulate the tumor invasion scenario with single-cell trackable analysis. To decode regulation during metastasis, we conducted in situ recovering for quantitative polymerase chain reaction (qPCR) analysis from each cell type from tumor-fibroblast copairing. Regulation of several essential genes has unveiled that matrix degradation gene MMP2 and angiogenesis associated gene VEGFA were up-regulated in tumor cells in the fibroblast-enriched niche compared with homogeneous cultivation. Therefore, this approach constitutes a novel tool for investigating metastasis with quantitative measurements both on phenotype and genetical information.


Asunto(s)
Dispositivos Laboratorio en un Chip , Metástasis de la Neoplasia/fisiopatología , Neoplasias/fisiopatología , Animales , Comunicación Celular/fisiología , Técnicas de Cultivo de Célula/métodos , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Movimiento Celular/genética , Movimiento Celular/fisiología , Fibroblastos/fisiología , Humanos , Técnicas Analíticas Microfluídicas/instrumentación , Técnicas Analíticas Microfluídicas/métodos , Metástasis de la Neoplasia/genética , Neoplasias/genética , Ratas
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