Integrating Reliable Pt-S Bond-Mediated 3D DNA Nanomachine with Magnetic Separation in a Homogeneous Electrochemical Strategy for Exosomal MicroRNA Detection with Low Background and High Sensitivity.

Precise and sensitive analysis of exosomal microRNA (miRNA) is of great importance for noninvasive early disease diagnosis, but it remains a great challenge to detect exosomal miRNA in human blood samples because of their small size, high sequence homology, and low abundance. Herein, we integrated reliable Pt-S bond-mediated three-dimensional (3D) DNA nanomachine and magnetic separation in a homogeneous electrochemical strategy for the detection of exosomal miRNA with low background and high sensitivity. The 3D DNA nanomachine was easily prepared via a facile and rapid freezing method, and it was capable of resisting the influence of biothiols, thus endowing it with high stability. Notably, the as-developed magnetic 3D DNA nanomachine not only enabled the detection system to have a low background but also coupled with liposome nanocarriers to synergistically amplify the current signal. Consequently, by ingeniously combining the low background and multiple signal-amplification strategies in homogeneous electrochemical biosensing, highly sensitive detection of exosomal miRNA was successfully achieved. More significantly, with good anti-interference ability, the as-proposed method could effectively discriminate plasma samples from cancer patients and healthy subjects, thus showing a high potential for application in the nondestructive early clinical diagnosis of disease.

Freeform optical system design with differentiable three-dimensional ray tracing and unsupervised learning.

Optical systems have been crucial for versatile applications such as consumer electronics, remote sensing and biomedical imaging. Designing optical systems has been a highly professional work due to complicated aberration theories and intangible rules-of-thumb, hence neural networks are only coming into this realm until recent years. In this work, we propose and implement a generic, differentiable freeform raytracing module, suitable for off-axis, multiple-surface freeform/aspheric optical systems, paving the way toward a deep learning-based optical design method. The network is trained with minimal prior knowledge, and it can infer numerous optical systems after a one-time training. The presented work unlocks great potential for deep learning in various freeform/aspheric optical systems, and the trained network could serve as an effective, unified platform for generating, recording, and replicating good initial optical designs.

Generalization of learned Fourier-based phase-diversity wavefront sensing.

Proper initialization of the nonlinear optimization is important to avoid local minima in phase diversity wavefront sensing (PDWS). An effective neural network based on low-frequency coefficients in the Fourier domain has proved effective to determine a better estimate of the unknown aberrations. However, the network relies significantly on the training settings, such as imaging object and optical system parameters, resulting in a weak generalization ability. Here we propose a generalized Fourier-based PDWS method by combining an object-independent network with a system-independent image processing procedure. We demonstrate that a network trained with a specific setting can be applied to any image regardless of the actual settings. Experimental results show that a network trained with one setting can be applied to images with four other settings. For 1000 aberrations with RMS wavefront errors bounded within [0.2 λ, 0.4 λ], the mean RMS residual errors are 0.032 λ, 0.039 λ, 0.035 λ, and 0.037 λ, respectively, and 98.9% of the RMS residual errors are less than 0.05 λ.

Tuning the magnetic ordering driven by cationic antisite defects in the Li(ZnMn)As system.

The electronic structure and magnetic properties of Li(ZnMn)As with antisite defects have been investigated by using first-principles calculations within the Perdew-Burke-Ernzerhof generalized gradient approximation. The cation antisite defect induced by Zn substitution for As was considered. Mn-3d, As-4p, Zn-4s, and Zn-4p were involved in the formation of d-sp hybrid orbitals, which enhanced the non-localized properties of Mn-3d electrons and provided a channel of Mn(↑)-As(↓)-ZnAs(↓)-Mn(↑) for indirect exchange of electrons between the magnetic ions. The antisite defect of Zn-substituted As belonged to the acceptor doping, rendering the compound p-type characteristics. The existence of the extra free hole carriers regulated the magnetic ordering transition. The ferromagnetic coupling between the Mn magnetic dopants was more favorable in the system with an antisite defect. In this paper, a novel type of dilute magnetic semiconductor with controllable carriers was designed and the mechanism of ferromagnetic coupling was revealed, which provided a theoretical reference for the subsequent studies.

Phase-diversity wavefront sensing enhanced by a Fourier-based neural network.

Phase diversity wavefront sensing (PDWS) has been a successful approach to quantifying wavefront aberrations with only a few intensity measurements and nonlinear optimization. However, the inherent non-convexity of the inverse problem may lead to stagnation at a local minimum far from the true solution. Proper initialization of the nonlinear optimization is important to avoid local minima and improve wavefront retrieval accuracy. In this paper, we propose an effective neural network based on low-frequency coefficients in the Fourier domain to determine a better estimate of the unknown aberrations. By virtue of the proposed network, only a small amount of simulation data suffice for a robust training, two orders of magnitude less than those in existing work. Experimental results show that, when compared with some existing methods, our method achieves the highest accuracy while drastically reducing the training time to 1.4 min. The minimum, maximum, and mean values of the root mean square (RMS) residual errors for 800 aberrations are 0.017λ, 0.056λ, and 0.039λ, respectively, and 95% of the RMS residual errors are less than 0.05λ.

Improvement of irradiation-induced fibroblast damage by α2-macroglobulin through alleviating mitochondrial dysfunction.

CONTEXT: α2-Macroglobulin (α2-M) is believed to be a potential anti-irradiation agent, but related mechanisms remains unclear. OBJECTIVE: We investigated the irradiation protective effect of α2-M. MATERIALS AND METHODS: A total of 10 Gy dose of irradiation was used to damage human skin fibroblasts. The influence of α2-M (100 µg/mL) on the proliferation, migration, invasion and apoptosis of fibroblasts was observed using Cell Counting Kit-8 (CCK8), wound healing, transwell, and flow cytometry. Malondialdehyde, superoxide dismutase and catalase was measured using related ELISA kits. The levels of mitochondrial membrane potential and calcium were detected using flow cytometry. The expression of transient receptor potential melastatin 2 (TRPM2) was investigated through western blotting and immunofluorescence staining. RESULTS: High purity of α2-M was isolated from Cohn fraction IV. α2-M significantly increased cell proliferation, migration, invasion, but suppressed cell apoptosis after irradiation. The promotion of cell proliferation, migration and invasion by α2-M exceeded over 50% compared group irradiation. The increased cell ratio in the S phase and decreased cell ratio in the G2 phase induced by irradiation were remarkably reversed by α2-M. α2-M markedly suppressed the increased oxidative stress level caused by irradiation. The mitochondrial damage induced by irradiation was improved by α2-M through inhibiting mitochondrial membrane potential loss, calcium and TRPM2 expression. DISCUSSION AND CONCLUSIONS: α2-M significantly promoted the decreased fibroblast viability and improved the mitochondria dysfunction caused by irradiation. α2-M might present anti-radiation effect through alleviating mitochondrial dysfunction caused by irradiation. This study could provide a novel understanding about the improvement of α2-M on irradiation-induced injury.

Identification of human parvovirus 4 genotypes 1 and 2 in Chinese source plasma pools.

Human parvovirus B19 (B19V) and human parvovirus 4 (PARV4) are known to infect humans and transmit through contaminated blood and blood products. Globally, three genotypes of B19V, as well as PARV4, have been identified, respectively. The existence of different B19V genotypes in Chinese plasma donors has been investigated, however, the data regarding PARV4 were not available. The main objective of this study is to identify the genotypes of PARV4 circulating in Chinese plasma donors. By using a duplex quantitative polymerase chain reaction assay adapted for all genotypes of B19V and PARV4, 78 source plasma pools for fractionation were screened and quantified. Results showed that positive rates of B19V and PARV4 DNA in plasma pool samples were 25.64% and 14.10%, respectively. PARV4 sequences in two positive samples were next genotyped, and these two sequences belonged to PARV4 genotypes 1 and 2, respectively. In conclusion, the data present demonstrate the existence of PARV4 genotypes 1 and 2 in Chinese plasma donors for the first time and also show the relatively lower prevalence and level of PARV4 DNA in Chinese plasma donors in comparison with that of B19V DNA.

Analysis of Protein Expression in Human Cells Cocultured with Porcine Peripheral Blood Mononuclear Cells.

OBJECTIVE: Porcine endogenous retroviruses (PERV) involved in pig to human xenotransplantation have raised great concerns because of their ubiquitous nature in pigs and their ability of infecting human cells in vitro. Although no significant cytopathic effect attributed to PERV was evident on PERV-infected human embryonic kidney 293 (HEK293) cells, we did proteomic analysis to investigate the differences of protein profile in order to further characterize the effect of PERV infection. METHODS: HEK293 cells were cocultured with porcine peripheral blood mononuclear cells (PBMCs). Protein profiles of PERV-infected and -noninfected HEK293 cells were analyzed by two-dimensional gel electrophoresis (2-DE). Protein spots with at least 1.5-fold alteration were identified by high-definition mass spectrometry (HDMS) analysis. Then real-time RT-PCR and Western blotting were performed to validate the proteomic results. RESULTS: Differential analysis of PERV-infected and -noninfected HEK293 cells by 2-DE revealed ten differentially regulated proteins. The proteins identified by HDMS were involved in various cellular pathways including signal transduction, cell apoptosis, and protein synthesis. CONCLUSION: The results of this study revealed differentially expressed proteins in HEK293 cells cocultured with porcine PBMCs and implied that these changes were probably induced by PERV infection. These results provide clues and potential links to understanding the molecular effect of the infection by human-tropic PERV.

Large-scale purification of high purity α1-antitrypsin from Cohn Fraction IV with virus inactivation by solvent/detergent and dry-heat treatment.

α1-Antitrypsin (AAT) is widely used to treat patients with congenital AAT deficiency. Cohn Fraction IV (Cohn F IV) is normally discarded during the manufacturing process of albumin but contains approximately 33% of plasma AAT. We established a new process for large-scale purification of AAT from it. liquid chromatography-electrospray ionization-tandem mass spectrometry and high-performance liquid chromatography were applied for qualitative identification and composition analysis, respectively. Stabilizers were optimized for AAT activity protection during lyophilization and dry-heat. Virus inactivation by dry-heat and solvent/detergent (S/D) was validated on a range of viruses. AAT with purity of 95.54%, specific activity of 3,938.5 IU/mg, and yield of 26.79%, was achieved. More than 95% activity was reserved after S/D. More than 96% activity was obtained after lyophilization or dry-heat. After S/D, pseudorabies virus (PRV) and vesicular stomatitis virus (VSV) were inactivated below detectable level within 1 H. Virus titer reductions of more than 5.50 log10 and 5.38 log10 were achieved for PRV and VSV, respectively. Porcine parvovirus and encephalomyocarditis virus were inactivated by 3.17 log10 and 5.88 log10 reduction after dry-heat. The advantages of this process, including suitability for large-scale production, high purity, better utilization of human plasma, viral safety, commercial and inexpensive chromatography medium, may facilitate its further application.

Assessment of lymph node metastasis in elderly patients with colorectal cancer by sentinel lymph node identification using carbon nanoparticles.

PURPOSE: To evaluate the value of sentinel lymph node (SLN) identification using carbon nanoparticles in abdominal lymph node metastasis in elderly (>60 years old) patients with colorectal cancer. METHODS: Eighty patients admitted at Weihai Second Municipal Hospital affiliated to Qingdao University from November 2014 to February 2017 were selected and divided into the control group (n=40) and the observation group (n=40) using the random number method. The control group was treated with surgery, while the observation group was administered carbon nanoparticle tracer for intraoperative dye detection and positioning; the first to four black-stained lymph nodes were marked as SLN, then radical surgery for colorectal cancer was performed. Pathological examination of intraoperative specimens was performed to assess the effect of SLN in the abdominal lymph node metastasis. RESULTS: There were no statistically significant differences in the metastasis rate and lymph node metastasis rate between the two groups (p>0.05). The total number of lymph nodes and the number of lymph nodes with micrometastases (<2mm) in the observation group were larger than those in the control group (p<0.05); the ratio of fewer than 12 lymph nodes in the observation group was lower than that in the control group (p<0.05). In the observation group, 8 out of 40 cases had lymph node metastasis, the detection rate of SLN using carbon nanoparticles was 92.50%, the accuracy rate 94.59%, the specificity of diagnosis 87.50%, the false negative rate 12.50% and the negative predictive value 21.88%. There was no statistically significant difference in the metastasis rate of black-stained and non-black-stained lymph nodes in the observation group (p>0.05). The blackstained rate of micro lymph nodes was higher than the total black-stained rate (p<0.05); the rate of micro lymph node metastasis was lower than that of lymph node metastasis >5mm (p<0.05). CONCLUSION: Preoperative SLN examination can evaluate the abdominal lymph node status in elderly patients with colorectal cancer, which is simple and accurate and can guide the clinical treatment, so it is worthy of popularization and application.

Assessment of lymph node metastasis in elderly patients with colorectal cancer by sentinel lymph node identification using carbon nanoparticles

PURPOSE: To identify the sentinel lymph node (SLN) in abdominal lymph node metastases of elderly patients with colorectal cancer using carbon nanoparticles (CN). METHODS: 80 colorectal cancer patients admitted to the affiliated Weihai Second Municipal Hospital of Qingdao University from November 2014 to February 2017 were selected, and divided into the control group (n=40) and the study group (n=40). The control group was treated with surgery, while the study group was administered CN tracer subcutaneously for intraoperative dye positioning; the first to four black-stained lymph nodes were marked as SLN and then radical surgery for CRC followed. Pathological examination of intraoperative specimens was performed to assess SLN metastasis. RESULTS: There were no statistically significant differences in the distant metastasis rate and SLN metastasis rate between the two groups (p>0.05). The total number of lymph nodes and the number of micro lymph nodes (<2 mm) in the study group was higher compared with the control group (p<0.05); the ratio of <12 lymph nodes in the study group was lower compared with the control group (p<0.05). In the study group, 8 out of 40 cases had SLN metastasis, the detection rate of SLN using CN was 92.50%, the accuracy rate was 94.59%, the specificity of diagnosis was 87.50%, the false negative rate was 12.50% and the negative predictive value was 21.88%. Νo statistically significant difference was noted in the metastasis rate of black-stained lymph nodes and non-black-stained lymph nodes in the study group (p>0.05). The black-stained rate of micro lymph nodes was higher than the total black-stained rate (p<0.05). Τhe rate of micro lymph node metastasis was lower than that of lymph node metastasis >5mm (p<0.05). CONCLUSION: Preoperative SLN examination can evaluate the abdominal lymph node status in elderly patients with colorectal cancer, which is simple and accurate and can guide the clinical treatment, so it is worthy of popularization and application.

Simultaneous detection and differentiation of human parvovirus B19 and human parvovirus 4 by an internally controlled multiplex quantitative real-time PCR.

Human parvovirus B19 (B19V) and human parvovirus 4 (PARV4) are two parvoviruses known to infect humans and transmit through blood and plasma derived medicinal products (PDMPs). Inactivation of the two parvoviruses has proven to be difficult and nucleic acid testing (NAT) would be an efficient means to exclude viruses. In this study, an internally controlled multiplex quantitative real-time PCR (qPCR) assay for B19V and PARV4 simultaneous detection and quantification was established and evaluated. The optimized multiplex qPCR assay allowed for simultaneous detection of all of the genotypes (1-3) of B19V and PARV4, with equal limit of quantification (LOQ) of 5 copies/µL, rather than other blood-borne viruses. It had a wide dynamic range of reliable amplification linearity of at least 8 orders of magnitude. Low standard deviations (SD) of quantification cycle (Cq) values and low coefficients of variation (CV) of copy numbers for both B19V and PARV4 suggested a high level of repeatability and reproducibility for the multiplex qPCR assay. This multiplex qPCR assay can be served as a readily applicable approach to screen plasma units intended for further manufacturing into PDMPs to reduce the risk of parvoviruses infection by such products and may also be useful for the detection of B19V/PARV4 co-infection or co-existence.

Inactivation of viruses by pasteurization at 60 °C for 10 h with and without 40% glucose as stabilizer during a new manufacturing process of α2-Macroglobulin from Cohn Fraction IV.

Pasteurization is regularly used to inactivate viruses for the safety of plasma derivatives. Influence of pasteurization at 60 °C for 10 h on α2-Macroglobulin activity and virus inactivation were studied. With 40% sugar as stabilizers more than 70% α2-Macroglobulin activity was reserved after pasteurization compared with 20% in control. Glucose presented a better activity protection effect than sucrose and maltose. By pasteurization without stabilizer the virus titers of pseudorabies virus, Sindbis virus, porcine parvovirus and encephalomyocarditis virus were reduced more than 5.88 log10, 7.50 log10, 4.88 log10, and 5.63 log10 respectively within 2 h. By pasteurization with 40% glucose vesicular stomatitis virus was inactivated more than 5.88 log10 within 1 h. Only 2.71 log10 reduction was achieved for encephalomyocarditis virus after 10 h. 40% glucose protected α2-M activity and viruses simultaneously from pasteurization. Other viral inactivation methods need to be incorporated to ensure viral safety of this manufacturing process of α2-Macroglobulin.

Inactivation of viruses during a new manufacturing process of α2-macroglobulin from Cohn Fraction IV by dry-heat treatment.

BACKGROUND: α2-Macroglobulin (α2-M) has a curative effect on radiation injury. Virus transmission through plasma derivatives is still not risk-free. Effect of dry heat on α2-M activity and virus inactivation by dry heat in a new manufacturing process of α2-M were studied. STUDY DESIGN AND METHODS: Effects of 100°C for 30 minutes, 80°C for 72 hours, and lyophilization on α2-M activity were detected, and stabilizing agents were optimized. Effect of a treatment at 100°C for 30 minutes has been tested on a range of viruses and characteristics change of α2-M was investigated. RESULTS: More than 90 and 80% α2-M activity recovery were reserved after treatment at 100°C for 30 minutes and 80°C for 72 hours, respectively. A concentration of 0.05 mol/L histidine presented a better protecting effect for α-M activity. No substantial changes were observed in the characteristics of α2-M compared with the untreated. By lyophilization and dry-heat treatment at 100°C for 30 minutes, murine encephalomyocarditis virus and pseudorabies virus (PRV) were inactivated below detectable level within 5 minutes (virus titers reduction ≥ 5.75 log) and 30 minutes (virus titers reduction ≥ 6.00 log), respectively. Bovine viral diarrhea virus and porcine parvovirus were inactivated by 4.29 and 2.46 log reduction, respectively. CONCLUSION: Treatment at 100°C for 30 minutes could improve the virus safety of α2-M with a slight activity loss.

Existence of various human parvovirus B19 genotypes in Chinese plasma pools: genotype 1, genotype 3, putative intergenotypic recombinant variants and new genotypes.

BACKGROUND: Human parvovirus B19 (B19V) is a frequent contaminant of blood and plasma-derived medicinal products. Three distinct genotypes of B19V have been identified. The distribution of the three B19V genotypes has been investigated in various regions or countries. However, in China, data on the existence of different B19V genotypes are limited. METHODS: One hundred and eighteen B19V-DNA positive source plasma pool samples collected from three Chinese blood products manufacturers were analyzed. The subgenomic NS1/VP1u region junction of B19V was amplified by nested PCR. These amplified products were then cloned and subsequently sequenced. For genotyping, their phylogenetic inferences were constructed based on the NS1/VP1-unique region. Then putative recombination events were analyzed and identified. RESULTS: Phylogenetic analysis of 118 B19V sequences attributed 61.86 % to genotype 1a, 10.17 % to genotype 1b, and 17.80 % to genotype 3b. All the genotype 3b sequences obtained in this study grouped as a specific, closely related cluster with B19V strain D91.1. Four 1a/3b recombinants and 5 new atypical B19V variants with no recombination events were identified. CONCLUSIONS: There were at least 3 subtypes (1a, 1b and 3b) of B19V circulating in China. Furthermore, putative B19V 1a/3b recombinants and unclassified strains were identified as well. Such recombinant and unclassified strains may contribute to the genetic diversity of B19V and consequently complicate the B19V infection diagnosis and NAT screening. Further studies will be required to elucidate the biological significance of the recombinant and unclassified strains.

Establishment of evanescent wave fiber-optic immunosensor method for detection bluetongue virus.

The evanescent wave fiber immunosensors (EWFI) technique was developed for the real-time rapidly sensitive and specific detection of the monoclonal antibody 3E2 of BTV. The outer-core protein VP7 of BTV was labled on the surface of the exposed fiber-optic core. The monoclonal antibody 3E2 of BTV VP7 were added and then the goat ant-rat IgG conjugated with Cy3 was captured. After the 532nm pulse (excitation source) reached the fiber probe, evanescent wave was generated, which excited the Cy3 bound to the immuno-complex and produced the fluorescent signal, which was changed into electrical signals read through computer. The preliminary results suggested that a detection limit of 10ng/ml was measured for the monoclonal antibody 3E2, which is equal to the sensitivity of ELISA. The 3E2 sample was specifically detected through the EWFI assay in 15min, and the fiber can be recycled at least ten times through TEA solution condition. This developed EWFI was a real-time rapidly sensitive and specific way for the detection of BTV antibodies.

Development of novel application of 3,3'-diindolylmethane: sensitizing multidrug resistance human breast cancer cells to γ-irradiation.

CONTEXT: Multidrug resistance (MDR) is known as a major obstacle to effective cancer therapy. The effects of irradiation on MDR in cancer cells had rarely been reported. OBJECTIVE: The effect of 3,3'-diindolylmethane (DIM) sensitizing MDR human breast carcinoma to γ-irradiation was investigated. MATERIALS AND METHODS: MCF-7/ADR cells were exposed to different concentrations of DIM (0-30 µM) for 48 or 2 h before IR (γ-Co60, 10 Gy, room temperature) then cultured for 48 h. Cell survival was determined by MTT assay. Intracellular reactive oxygen spices (ROS) induced by DIM (20 and 30 µM, 2 h before irradiation) was measured by flow cytometry. Propidium iodide staining assay was used for cell cycle distribution studies; cell apoptosis was measured by flow cytometry and confocal microscopy. RESULTS: DIM (20 and 30 µM, 2 h before irradiation) sensitized MCF-7/ADR cells to IR with survival rates decreased from 100% to 79% and 63%, respectively. DIM combined with γ-radiation demonstrated that the activity of G2/M phase cell cycle arresting with percentages enhanced from 9% to 49% and 52%. DIM can increase intracellular ROS generation by 1.45- and 1.55-times compared to control group. Significantly enhanced radiation-induced apoptosis by DIM was also observed. DISCUSSION AND CONCLUSION: These data provide a rationale for the use of DIM as a promising radio-sensitizer to MDR cancer cells.

Infection of common marmosets with hepatitis C virus/GB virus-B chimeras.

UNLABELLED: The development of vaccination and novel therapy for hepatitis C virus (HCV) has been hampered by the lack of suitable small-animal models. GB virus B (GBV-B), closely related to HCV, causes viral hepatitis in common marmosets (Callithrix jacchue jacchus) and might represent an attractive surrogate model for HCV infection. However, differences exist between GBV-B and HCV in spite of a short genetic distance between the two viruses. Here we report common marmosets infected with two HCV/GBV-B chimeras containing HCV structural genes coding for either whole core and envelope proteins (CE1E2p7) or full envelope proteins (E1E2p7) substituted for the counterpart elements of GBV-B. Naïve animals intrahepatically injected with chimeric RNA transcripts or intravenously injected with sera from primary infected animals produced high levels of circulating infectious chimeric viruses and they developed chronic infection. Tacrolimus-treated marmosets inoculated with a CE1E2p7 chimera had higher viral loads and long-term persistent infection. A moderate elevation of serum aspartate aminotransferase (AST) levels was observed in parallel with viral replication. Chimeras recovered from liver samples revealed 1/958 adaptive viral mutations. Histopathological changes typical of viral hepatitis were observed in liver tissues from all types of HCV chimeras-infected marmosets. HCV core and E2 proteins were detected in liver tissues from infected animals by immunohistochemical staining. Fluctuations of chimeric virus replication in marmosets with spontaneous and sporadic viral clearance might be related to specific antibody and T-cell response to HCV proteins in vivo. Replication of CE1E2p7 chimera was observed in primary hepatocyte cultures by immunofluorescent staining in vitro. CONCLUSION: Infectious HCV chimeras causing chronic hepatitis in marmosets might constitute a small primate model suitable for evaluation of virus-cell interaction, vaccination, and antiviral therapy against HCV infection.

MicroRNA-125b promotes tumor metastasis through targeting tumor protein 53-induced nuclear protein 1 in patients with non-small-cell lung cancer.

BACKGROUND: Lung cancer, predominantly non-small-cell lung cancer (NSCLC), is the leading cause of cancer deaths worldwide. There is a great need to identify critical effectors involved in metastasis of NSCLC that will facilitate the development of new therapeutic strategies. Here we evaluated the potential role of miR-125b in the metastasis of NSCLC cells. METHODS: Human NSCLC cells were isolated from surgical tissues with Cancer Cell Isolation Kit. Expressions of miR-125b and TP53INP1 were detected with real-time PCR and western blot. Human miR-125b mimics, miR-125b inhibitor, TP53INP1 expression plasmid and TP53INP1 siRNA were transfected into NSCLC cells with nucleofector transfection kit. NSCLC metastasis was determined with adhesion assay, invasive assay and lung tumor metastasis model. RESULTS: The expression of miR-125b was significantly higher in poorly differentiated NSCLC cells that are endowed with high metastatic potentials. Up-regulation of miR-125b could enhance the metastatic potential of NSCLC cells in vitro and in vivo, while down-regulation of miR-125b resulted in decreased metastatic potentials in vitro and in vivo. Further, tumor protein 53-induced nuclear protein 1 (TP53INP1) was an important target of miR-125b involved in metastasis of NSCLC cells. TP53INP1 served as a negative regulator of NSCLC metastasis. Decreased expression of TP53INP1 in tumor tissues was inversely associated with their expression of miR-125b, significantly lower in poorly differentiated tumors and inversely correlated with the clinical stages in patients with NSCLC. CONCLUSIONS: These findings demonstrated that miR-125b promoted tumor metastasis via targeting TP53INP1 in human NSCLC cells, which uncovered a real clinical relevance of microRNAs in tumor biology, and provided novel potential candidates for NSCLC clinical practice.

The effectiveness of riboflavin photochemical-mediated virus inactivation and changes in protein retention in fresh-frozen plasma treated using a flow-based treatment device.

BACKGROUND: A flow-based treatment device using riboflavin and ultraviolet (UV) light was developed to inactivate viruses in fresh-frozen plasma (FFP). The objective of this study was to evaluate the in vitro effectiveness of virus inactivation and changes in protein quality in FFP treated with this device. STUDY DESIGN AND METHODS: FFP-contaminating viruses were treated with riboflavin and UV light using a one-pass linear flow device. The infectivity of viruses was measured using established biologic assays. Real-time polymerase chain reaction (PCR) was performed to detect damage to viral nucleotides after treatment. Treated plasma was analyzed using standard coagulation assays. RESULTS: FFP treated at the UV dose of 3.6 J/cm(2) (J) exhibited a mean reduction of virus titer of more than 4 logs. The effectiveness increased significantly at higher doses. Real-time PCR showed that the cycle threshold values for both complete inactivation and virus recultivation were higher than that of the untreated sample. At doses of 3.6, 5.4, and 7.2 J, the protein recovery rates were 60.2 ± 8.6, 46.6 ± 9.4, and 28.0 ± 1.0% for fibrinogen; 67.0 ± 3.1, 57.3 ± 8.0, and 49.2 ± 3.8% for Factor VIII; 93.6 ± 2.8, 89.6 ± 6.1, and 86.5 ± 5.3% for antithrombin-III; and 72.1 ± 5.6, 59.8 ± 14.2, and 49.2 ± 8.4% for Protein C, respectively. CONCLUSION: The effectiveness of virus inactivation was enhanced, but total activity of plasma factors was reduced, in a UV dose-dependent manner.