Precision Phenotypic Profiling and Capture of Circulating Tumor Cells via a Vertical Laminar Flow-Stacked Microfluidic Chip.

The heterogeneity of circulating tumor cells has a significant impact on the diagnosis, treatment, and monitoring of cancer. Research on the subtypes of circulating tumor cells can bring better treatment outcomes for cancer patients. Here, we proposed a microfluidic chip for the magnetic capture of subtypes of circulating tumor cells from the whole blood and phenotypic profiling by stacking laminar flow vertically. Circulating tumor cells were sorted and captured by the three-dimensional regulation of both magnetic fields in the vertical direction and flow fields in the lateral direction. Using EpCAM-magnetic beads, we achieved sorting and sectional capture of target cells in whole blood and analyzed the surface expression levels of the captured cells, confirming the functionality of the microfluidic chip in sorting and capturing subtypes of circulating tumor cells. This microfluidic chip can also aid in the subsequent subtype analysis of other rare cells.

Analysis of the Pattern Shapes Obtained By Micro/Nanospherical Lens Photolithography.

Micro/nanospherical lens photolithography (SLPL) constitutes an efficient and precise micro/nanofabrication methodology. It offers advantages over traditional nanolithography approaches, such as cost-effectiveness and ease of implementation. By using micrometer-sized microspheres, SLPL enables the preparation of subwavelength scale features. This technique has gained attention due to its potential applications. However, the SLPL process has a notable limitation in that it mostly produces simple pattern shapes, mainly consisting of circular arrays. There has been a lack of theoretical analysis regarding the possible shapes that can be created. In our experiments, we successfully prepared annular and ring-with-hole pattern shapes. To address this limitation, we applied the Mie scattering theory to systematically analyze and summarize the various patterns that can be obtained through the SLPL process. We also proposed methods to predict and obtain different patterns. This theoretical analysis enhances the understanding of SLPL and expands its potential applications, making it a valuable area for further research.

Manipulation of Particle/Cell Based on Compressibility in a Divergent Microchannel by Surface Acoustic Wave.

The mechanical properties (compressibility or deformability) of cells are closely related to their death, migration, and differentiation. Accurate separation and manipulation of bioparticles based on these mechanical properties are still a challenging in the field of acoustofluidics. In this work, based on surface acoustic waves (SAW) and divergent microchannels, we developed a new method for separating and detecting particles or cells with different compressibility. The difference in acoustic radiation force (Fr) caused by compressibility are gradually amplified and accumulated by decreasing the flow velocity, and they are finally reflected in the particle migration distance. During the transverse migration process, the alternating dominance of the acoustic radiation force and the Stokes resistance force (Fs) drives the particles to create three typical migration patterns: intermittent migration, compound migration, and near-wall migration. In the present tilted SAW device, a 91% separation success rate of ∼10 µm polystyrene (PS) and polydimethylsiloxane (PDMS) particles can be achieved by optimizing the acoustic field input power and the fluid velocity. The application potential of the present divergent microchannel is validated by separating the myelogenous leukemia cell K562 and the natural killer cell NK92 that have similar densities and sizes (∼15 µm) but different compressibility. The results of this work are expected to provide valuable insights into the acoustofluidics separation and detection of the cells that are with different compressibility.

High-Throughput and Efficient Intracellular Delivery Method via a Vibration-Assisted Nanoneedle/Microfluidic Composite System.

Intracellular delivery and genetic modification have brought a significant revolutionary to tumor immunotherapy, yet existing methods are still limited by low delivery efficiency, poor throughput, excessive cell damage, or unsuitability for suspension immune cells, specifically the natural killer cell, which is highly resistant to transfection. Here, we proposed a vibration-assisted nanoneedle/microfluidic composite system that uses large-area nanoneedles to rapidly puncture and detach the fast-moving suspension cells in the microchannel under vibration to achieve continuous high-throughput intracellular delivery. The nanoneedle arrays fabricated based on the large-area self-assembly technique and microchannels can maximize the delivery efficiency. Cas9 ribonucleoprotein complexes (Cas9/RNPs) can be delivered directly into cells due to the sufficient cellular membrane nanoperforation size; for difficult-to-transfect immune cells, the delivery efficiency can be up to 98%, while the cell viability remains at about 80%. Moreover, the throughput is demonstrated to maintain a mL/min level, which is significantly higher than that of conventional delivery techniques. Further, we prepared CD96 knockout NK-92 cells via this platform, and the gene-edited NK-92 cells possessed higher immunity by reversing exhaustion. The high-throughput, high-efficiency, and low-damage performance of our intracellular delivery strategy has great potential for cellular immunotherapy in clinical applications.

[Retracted] Long intergenic non­protein coding RNA 152 promotes multiple myeloma progression by negatively regulating microRNA­497.

Following the publication of this paper, an interested reader drew the authors' attention to the fact that several of the tumour images shown in Fig. 6B were strikingly similar to images that had appeared in another paper by different authors published at around the same time. The authors also admitted that the in vivo experiments reported in this study had been performed by a third party. Consequently, owing to a lack of confidence in the presented data, the authors requested that this paper be retracted from the Journal. The Editor was in agreement that the paper should be retracted. All authors agree with the retraction of this article, and the Editor apologizes to the readership for any inconvenience caused. [the original article was published in Oncology Reports 40: 3763­3771, 2018; DOI: 10.3892/or.2018.6721].

Clinicopathological characteristics and prognostic factors for intrahepatic cholangiocarcinoma: a population-based study.

We aimed to explore the clinicopathological features and survival-related factors for intrahepatic cholangiocarcinoma (ICC). Eligible data were extracted from the Surveillance, Epidemiology and End Results (SEER) database from 2004 to 2015. Totally, 4595 ICC patients were collected with a male to female ratio of nearly 1:1. The higher proportion of ICC patients was elderly, tumor size ≥ 5 cm and advanced AJCC stage. Most patients (79.2%) have no surgery, while low proportion of patients receiving radiotherapy (15.1%). The median survival was 7.0 months (range 0-153 months). The 5-year CSS and OS rates were 8.96% and 7.90%. Multivariate analysis found that elderly age (aged ≥ 65 years old), male, diagnosis at 2008-2011, higher grade, tumor size ≥ 5 cm, and advanced AJCC stage were independent factors for poorer prognosis; while API/AI (American Indian/AK Native, Asian/Pacific Islander) race, married, chemotherapy, surgery and radiotherapy were independent favorable factors in both CSS and OS. Furthermore, stratified analysis found that chemotherapy and radiotherapy improved CSS and OS in patients without surgery. Age, sex, race, years of diagnosis, married status, grade, tumor size, AJCC stage, surgery, chemotherapy and radiotherapy were significantly related to prognosis of ICC. Chemotherapy and radiotherapy could significantly improve survival in patients without surgery.

Improved polarization and energy density in boron nitride nanosheets/poly(vinylidene fluoride-chlorotrifluoroethylene) nanocomposite with trilayered architecture regulation.

A polymer film capacitor with high energy density has attracted great attention in recent decades to fulfil the requirements of miniaturization and light weight for electronics and energy storage systems. The multi-layered nanocomposite exhibits an effective route for developing polymer film with large energy capability due to the advantage of each layer. Here, two sandwich constructions of boron nitride nanosheets (BNNSs)/poly(vinylidene fluoride-chlorotrifluoroethylene) (P(VDF-CTFE)) nanocomposite film were designed to investigate the effects of architecture regulation on energy storage performance, i.e. the nanocomposite serving as an inner layer with pristine films as outer layers, in which the nanocomposite film delivers a strongly polarized behavior and the pristine polymer provides a high breakdown strength. The BNNSs were liquid exfoliated from boron nitride bulk assisted with fluoro hyperbranched copolymer that was adsorbed on the surface of nanosheets via CH-π non-covalent interactions. The presence of fluoro segments improves the compatibility between the nanosheets and the P(VDF-CTFE) matrix. The maximum polarization of 7.9 µC cm-2 at 300 MV m-1 is achieved in the tailored sandwich film, and the discharged energy density of 9.1 J cm-3 is obtained in the current nanocomposite, which benefits from the barrier effect between adjacent layers induced by the redistribution of electric field and emerging charge traps at the interfaces. This sandwich BNNSs/P(VDF-CTFE) nanocomposite provides an insightful prospect for structural design with respect to the advanced application of the polymer film capacitor.

Efficient exfoliation of UV-curable, high-quality graphene from graphite in common low-boiling-point organic solvents with a designer hyperbranched polyethylene copolymer and their applications in electrothermal heaters.

The use of stabilizer with designer structures can effectively promote graphite exfoliation in common solvents to render functionalized graphene desirable for their various applications. Herein, a hyperbranched polyethylene copolymer, HBPE@Py@Acryl, simultaneously bearing multiple pyrene terminal groups and pendant acryloyl moieties has been successfully synthesized from ethylene with a Pd-diimine catalyst based on unique chain walking mechanism. The unique structural design of the HBPE@Py@Acryl makes it capable of effectively promote graphite exfoliation in a series of common, low-boiling-point organic solvents, e.g. CHCl3, to render stable graphene dispersions with concentrations effectively adjustable by changing feed concentrations of graphite and polymer or sonication time. Meanwhile, it can be irreversibly adsorbed on the exfoliated graphene surface based on the π-π interactions between them to concurrently render acryloyl-functionalized graphene free of structural defects, with majority (92.7%) of them having a thickness of 2-3 layers. This allows us to obtain graphene electrothermal films simply by filtration and UV irradiation, which exhibit outstanding stability in use. The action mechanism of the HBPE@Py@Acryl as stabilizer for promoting graphite exfoliation and the role of UV irradiation on improving the stability in use of resulting graphene films have been elucidated.

Enhanced interfacial polarization in poly(vinylidene fluoride-chlorotrifluoroethylene) nanocomposite with parallel boron nitride nanosheets.

The miniaturization of electronics provides an opportunity for the polymer film capacitor due to its lightweight and flexibility. In order to improve energy density and charge-discharge efficiency of the film capacitor, the development of a polymer nanocomposite is one of the effective strategies, in which the distribution of the fillers plays a key role in the enhancement of the electrical energy capability. In this work, the few-layer boron nitride nanosheets (BNNSs) was exfoliated with assistance of the fluoro hyperbranched polyethylene-graft-poly(trifluoroethyl methacrylate) (HBPE-g-PTFEMA) copolymer as stabilizer, which was adsorbed on the surface of the nanosheets via a CH-π non-covalent interaction. The morphological results confirm the lateral size of ∼0.4 µm for resultant nanosheets with the intact crystal structure. The loading of 0.5 vol% BNNSs was embedded into poly(vinylidene fluoride-chlorotrifluoroethylene) (P(VDF-CTFE)) matrix by solution casting method, and then the nanocomposite film was uniaxial stretched to achieve the orientation of nanosheets in polymer host. The dielectric constant of stretching nanocomposite with ratio of 4 at 50 mm min-1 reaches 51.1 at 100 Hz with low loss as 0.016, while the energy density of 7.0 J cm-3 at 250 MV m-1 with charge-discharge efficiency of 56% is obtained in current nanocomposite film, which is attributed to the interfacial polarization as well as parallel nanosheets blocking the growth of electrical treeing branches. This strategy of the aligned nanosheets/polymer nanocomposite establishes a simple route to construct heterogeneity in polymer films with enhanced electrical energy capability for flexible capacitors.

Long-term effect of catheter ablation on tachycardia-bradycardia syndrome: evidenced by 10 years follow up.

Background: Catheter ablation has been used for the treatment of tachycardia-bradycardia syndrome (TBS). However, data on its long-term effect of rhythm control and stroke are limited.Method and results: Patients with TBS admitted in the First Affiliated Hospital of Dalian Medical University from 2002 to 2013 were reviewed in the present study. A total of 150 patients were enrolled. Seventy-nine patients underwent catheter ablation (CA group) and 71 patients chose implantation of pacemaker (PM group). The two groups were followed up for 123.01 ± 29.68 and 120.67 ± 31.05 months respectively. The CA group underwent 1.2 ± 0.5 procedure. Of the CA group, 70.9% patients exhibited sinus rhythm without long pauses or the need of anti-arrhythmia drugs (AADs). In contrast, no patient in the PM group was free of atrial fibrillation (AF). A higher proportion in the PM group progressed to persistent AF than in that in the CA group (9.9% vs. 1.3%, p < .05). The incidence of new-onset stroke in the PM group was significantly higher than that in the CA group (15.4% vs. 5.1%, p < .05).Conclusions: Even for long-term following up, catheter ablation is effective for preventing both the tachycardia and bradycardia components for the majority of patients with TBS without the need for further pacemaker implantation. Furthermore, ablation can reduce the stroke incidence of TBS through eliminating AF and reducing the progression to persistent AF.

Long non-coding RNA DSCAM-AS1 accelerates the progression of hepatocellular carcinoma via sponging miR-338-3p.

Aberrant expression of long non-coding RNA DSCAM-AS1 (Down Syndrome Cell Adhesion Molecule antisense) has been observed in several cancers. However, the expression status, biological function and underling mechanism of DSCAM-AS1 in hepatocellular carcinoma (HCC) remain unclear. The expression of DSCAM-AS1 was detected in HCC tissues and serum from both HCC patients and healthy controls. MTS, wound healing and transwell invasion assays were used to examine the effects of DSCAM-AS1 on cell proliferation, migration, and invasion in HCC cells, respectively. MicroRNAs (miRNAs) targeted DSCAM-AS1 was predicated by Starbase2.0 and identified using luciferase reporter and RNA immunoprecipitation assays. The xenograft mice were established to examine the effect DSCAM-AS1 on tumor growth in vivo. We found that DSCAM-AS1 was up-regulated in HCC tissues relative to adjacent non-tumor tissues. Serum levels of DSCAM-AS1 were higher in HCC patients than that in healthy controls. Increased DSCAM-AS1 was associated with poor prognosis. Knockdown of DSCAM-AS1 significantly inhibited HCC cell proliferation, migration and invasion. Moreover, miR-338-3p was confirmed as a direct target of DSCAM-AS1 in HCC cells. The miR-338-3p inhibitor could partially reverse the inhibitory effect of DSCAM-AS1 depletion in HCC cells. DSCAM-AS1 positively regulated CyclinD1 and smoothened (SMO) expression (two targets of miR-338-3p) in HCC cells. Moreover, tumor growth was tremendously retarded in nude mice received injection of SMCC-7721 cells transfected with sh-DSCAM-AS1. Taken together, the present work suggested that DSCAM-AS1 functioned as an oncogenic lncRNA that promoted HCC progression by sponging miR-338-3p.

Long intergenic non­protein coding RNA 152 promotes multiple myeloma progression by negatively regulating microRNA­497.

Long intergenic non­protein coding RNA 152 (LINC00152, also known as cytoskeleton regulator RNA) is reportedly involved in the development and progression of various types of human malignancy. However, the functional role of LINC00152 in multiple myeloma (MM) and the underlying molecular mechanisms have remained elusive. The present study aimed to investigate the role of LINC00152 in the genesis of MM and the potential molecular mechanisms. It was identified that the expression of LINC00152 was significantly upregulated in plasma cells from patients with MM vs. healthy subjects, and that a high expression of LINC00152 was correlated with a shorter overall survival in patients with MM. Functional assays demonstrated that knockdown of LINC00152 by transfecting MM cells with LINC00152­specific short hairpin RNA expression plasmids significantly inhibited cell proliferation by inducing cell cycle arrest at the G0/G1 phase. Furthermore, knockdown of LINC00152 promoted caspase­3/9 activity and apoptosis in MM cells. In addition, knockdown of LINC00152 significantly attenuated tumor growth in mice bearing a myeloma xenograft. A luciferase reporter assay indicated that microRNA (miR)­497 is a direct target of LINC00152, and its expression levels were inversely correlated with those of LINC00152 in MM tissues. Furthermore, repression of miR­497 partly abrogated the inhibitory effects of LINC00152 knockdown on MM cells. Collectively, the results indicated that LINC00152 has an oncogenic effect in MM by targeting miR­497, and may be a novel diagnostic marker and therapeutic target for MM.

Citrus aurantium Naringenin Prevents Osteosarcoma Progression and Recurrence in the Patients Who Underwent Osteosarcoma Surgery by Improving Antioxidant Capability.

Citrus aurantium is rich in flavonoids, which may prevent osteosarcoma progression, but its related molecular mechanism remains unclear. Flavonoids were extracted from C. aurantium and purified by reparative HPLC. Each fraction was identified by using electrospray ionisation mass spectrometry (ESI-MS). Three main components (naringin, naringenin, and hesperetin) were isolated from C. aurantium. Naringenin inhibited the growth of MG-63 cells, whereas naringin and hesperetin had no inhibitory function on cell growth. ROS production was increased in naringin- and hesperetin-treated groups after one day of culture while the level was always lowest in the naringenin-treated group after three days of culture. 95 osteosarcoma patients who underwent surgery were assigned into two groups: naringenin group (NG, received 20 mg naringenin daily, n = 47) and control group (CG, received 20 mg placebo daily, n = 48). After an average of two-year follow-up, osteosarcoma volumes were smaller in the NG group than in the CG group (P > 0.01). The rate of osteosarcoma recurrence was also lower in the NG group than in CG group. ROS levels were lower in the NG group than in the CG group. Thus, naringenin from Citrus aurantium inhibits osteosarcoma progression and local recurrence in the patients who underwent osteosarcoma surgery by improving antioxidant capability.

MicroRNA-497 suppresses cell proliferation and induces apoptosis through targeting PBX3 in human multiple myeloma.

Aberrant expression of microRNA-497 (miRN-497) is implicated in development and progression of multiple types of cancers. However, the biological function and underlying mechanism of miR-497 in multiple myeloma (MM) remains unclear. Thus, we studied the potential biological roles of miR-497 in MM. The expression of miR-497 was examined in multiple myeloma and normal plasma cells by qRT-PCR. Biological functions of miR-497 were analyzed using cell proliferation, colony formation, cell cycle, apoptosis and luciferase assays in vitro, as well as via tumorigenicity in vivo analysis. Here, we observed reduced expression of miR-497 in MM plasma samples and cell lines. Ectopic expression of miR-497 dramatically suppressed cell proliferation and clonogenicity, as well as induced cell arrest at G0/G1 stage and apoptosis in vitro. Mechanistic investigation assays showed that Pre-B-cellleukemia transcription factor 3 (PBX3) was a novel and direct downstream target of miR-497. Interestingly, overexpression of PBX3 partially reverted the effect of miR-497 in MM cells. In xenograft model, overexpression of miR-497 inhibited tumorigenicity by repressing PBX3. These findings collectively suggested that miR-497 functioned as tumor suppressor in MM by directly targeting PBX3, supporting its utility as a novel and potential therapeutic agent for MM therapy.