Evolution of multiple cell clones over a 29-year period of a CLL patient

Chronic lymphocytic leukaemia (CLL) is a frequent B-cell malignancy, characterized by recurrent somatic chromosome alterations and a low level of point mutations. Here we present single-nucleotide polymorphism microarray analyses of a single CLL patient over 29 years of observation and treatment, and transcriptome and whole-genome sequencing at selected time points. We identify chromosome alterations 13q14−, 6q− and 12q+ in early cell clones, elimination of clonal populations following therapy, and subsequent appearance of a clone containing trisomy 12 and chromosome 10 copy-neutral loss of heterogeneity that marks a major population dominant at death. Serial single-cell RNA sequencing reveals an expression pattern with high FOS, JUN and KLF4 at disease acceleration, which resolves following therapy, but reoccurs following relapse and death. Transcriptome evolution indicates complex changes in expression occur over time. In conclusion, CLL can evolve gradually during indolent phases, and undergo rapid changes following therapy

Full-length single-cell RNA-seq applied to a viral human cancer:applications to HPV expression and splicing analysis in HeLa S3 cells

Background: Viral infection causes multiple forms of human cancer, and HPV infection is the primary factor in cervical carcinomas Recent single-cell RNA-seq studies highlight the tumor heterogeneity present in most cancers, but virally induced tumors have not been studied HeLa is a well characterized HPV+ cervical cancer cell line Result: We developed a new high throughput platform to prepare single-cell RNA on a nanoliter scale based on a customized microwell chip Using this method, we successfully amplified full-length transcripts of 669 single HeLa S3 cells and 40 of them were randomly selected to perform single-cell RNA sequencing Based on these data, we obtained a comprehensive understanding of the heterogeneity of HeLa S3 cells in gene expression, alternative splicing and fusions Furthermore, we identified a high diversity of HPV-18 expression and splicing at the single-cell level By co-expression analysis we identified 283 E6, E7 co-regulated genes, including CDC25, PCNA, PLK4, BUB1B and IRF1 known to interact with HPV viral proteins Conclusion: Our results reveal the heterogeneity of a virus-infected cell line It not only provides a transcriptome characterization of HeLa S3 cells at the single cell level, but is a demonstration of the power of single cell RNA-seq analysis of virally infected cells and cancers

Influence of Thermally-Accelerated Aging on the Dynamic Mechanical Properties of HTPB Coating and Crosslinking Density-Modified Model for the Payne Effect

Hydroxyl terminated polybutadiene (HTPB) coating is widely used in a solid rocket motor, but an aging phenomenon exists during long-term storage, which causes irreversible damage to the performance of this HTPB coating. In order to study the effect of aging on the dynamic mechanical properties of the HTPB coating, the thermally-accelerated aging test was carried out. The variation of maximum elongation and crosslinking density with aging time was obtained, and a good linear relationship between maximum elongation and crosslinking density was found by correlation analysis. The changing regularity of dynamic mechanical properties with aging time was analyzed. It was found that with the increase of aging time, Tg of HTPB coating increased, Tα, tan β and tan α decreased, and the functional relationships between the loss factor parameters and crosslinking density were constructed. The storage modulus and loss modulus of HTPB coating increased with the increase of aging time, and decreased with the increase of pre-strain. The aging enhanced the Payne effect of HTPB coating, while the pre-strain had a weakening effect. In view of the Payne effect of HTPB coating, the crosslinking density was introduced into Kraus model as aging evaluation parameter, and the crosslinking density modified models with and without pre-strain were established. The proposed models can effectively solve the problem that the Kraus model has a poor fitting effect under the condition of small strain (generally less than 1%) and on the loss modulus, which have improved the correlations between the fitting results and the test results

Surgical Modification of the Murine Calvaria Osteolysis Model

The murine calvaria model has been adopted for evaluation of osteolysis and inflammation induced by polyethylene (PE) or metal wear debris. However, this model suffers from several complications. The purpose of our study is to introduce a surgical modification with lower complication rates, thus providing more accurate results. Forty C57/BL6 mice were divided into two groups, both receiving polyethylene particles. Surgical modifications were performed in group 1, and group 2 underwent traditional surgeries. The incidence of fluid leakage was recorded on the operative day. Curst formation, wound dehiscence, and bone exposure were recorded on day 7. Histological osteolysis was demonstrated by HE staining of tissue slices. Micro-CT was used for quantifying evaluation of osteolysis in two groups. Intraoperative fluid leakage was significantly reduced in group 1. Postoperative crust formation, wound dehiscence, and bone exposure were also significantly decreased in group 1. HE staining results revealed obvious osteolysis in group 1 and more obvious osteolysis in group 2. Bone volume fraction (BVF) was (0.32 ± 0.03) in group 1 compared to group 2 (0.24 ± 0.05). Bone mineral density (BMD) was (1.11 ± 0.03) in group 1 compared to group 2 (1.01 ± 0.02). Surgical modifications provide a reliable way for establishment of the murine calvaria osteolysis model

Enhanced Bounding Box Estimation with Distribution Calibration for Visual Tracking

Bounding box estimation by overlap maximization has improved the state of the art of visual tracking significantly, yet the improvement in robustness and accuracy is restricted by the limited reference information, i.e., the initial target. In this paper, we present DCOM, a novel bounding box estimation method for visual tracking, based on distribution calibration and overlap maximization. We assume every dimension in the modulation vector follows a Gaussian distribution, so that the mean and the variance can borrow from those of similar targets in large-scale training datasets. As such, sufficient and reliable reference information can be obtained from the calibrated distribution, leading to a more robust and accurate target estimation. Additionally, an updating strategy for the modulation vector is proposed to adapt the variation of the target object. Our method can be built on top of off-the-shelf networks without finetuning and extra parameters. It yields state-of-the-art performance on three popular benchmarks, including GOT-10k, LaSOT, and NfS while running at around 40 FPS, confirming its effectiveness and efficiency

Enhanced Bounding Box Estimation with Distribution Calibration for Visual Tracking

Bounding box estimation by overlap maximization has improved the state of the art of visual tracking significantly, yet the improvement in robustness and accuracy is restricted by the limited reference information, i.e., the initial target. In this paper, we present DCOM, a novel bounding box estimation method for visual tracking, based on distribution calibration and overlap maximization. We assume every dimension in the modulation vector follows a Gaussian distribution, so that the mean and the variance can borrow from those of similar targets in large-scale training datasets. As such, sufficient and reliable reference information can be obtained from the calibrated distribution, leading to a more robust and accurate target estimation. Additionally, an updating strategy for the modulation vector is proposed to adapt the variation of the target object. Our method can be built on top of off-the-shelf networks without finetuning and extra parameters. It yields state-of-the-art performance on three popular benchmarks, including GOT-10k, LaSOT, and NfS while running at around 40 FPS, confirming its effectiveness and efficiency

Morphologic changes within the cerebellar cortex in the unilateral 6-hydroxydopamine lesioned rat model for Parkinson disease

Parkinson’s disease (PD) is a common neurodegenerative disorder caused by the progressive loss of dopaminergic neurons in the substantia nigra. Most investigations have focused on the cerebral regions such as the basal ganglia, thalamus, or the substantia nigra, but whether there is pathologic impairment within the cerebellum has rarely been assessed. Synapsin and neurofilament as the inner markers of neurons and synapses reflect the functional state by their distribution or expression. Significant morphologic changes at the cellular level have been demonstrated directly or indirectly in multiple neurodegenerative diseases. The purpose of this study was to determine whether the behavioral abnormalities that accompany PD are associated with the cerebellum using an in vivo 6- hydroxydopamine lesioned rat model. Forty-two rats were divided into three groups, the Parkinsonian group (N=22), sham group (N=10) and control group (N=10). The dopaminergic lesion was determined by immunohistochemical analysis for tyrosine hydroxylaseimmunopositive cells. Immunohistochemical studies showed that the density of synapsin I in the granular layer of the cerebellum on both sides of the Parkinsonian -model was not statistically significantly different compared to the control and sham groups. However, expression of neurofilament H in the cortex within bilateral paramedian lobule (PML) and Crus 2 of the ansiform lobule (C2AL) in cerebellum posterior lobe of Parkinsonian rats was decreased compared with controls (P<0.05), especially in the loss of Purkinje cells and the presence of morphologic abnormalities in the cell nucleus. The study suggested that loss of neurons and synapses may take place in the cerebellar cortex of Parkinson’s disease, and might play an important role in the pathologic mechanism of PD

Surgical Modification of the Murine Calvaria Osteolysis Model

The murine calvaria model has been adopted for evaluation of osteolysis and inflammation induced by polyethylene (PE) or metal wear debris. However, this model suffers from several complications. The purpose of our study is to introduce a surgical modification with lower complication rates, thus providing more accurate results. Forty C57/BL6 mice were divided into two groups, both receiving polyethylene particles. Surgical modifications were performed in group 1, and group 2 underwent traditional surgeries. The incidence of fluid leakage was recorded on the operative day. Curst formation, wound dehiscence, and bone exposure were recorded on day 7. Histological osteolysis was demonstrated by HE staining of tissue slices. Micro-CT was used for quantifying evaluation of osteolysis in two groups. Intraoperative fluid leakage was significantly reduced in group 1. Postoperative crust formation, wound dehiscence, and bone exposure were also significantly decreased in group 1. HE staining results revealed obvious osteolysis in group 1 and more obvious osteolysis in group 2. Bone volume fraction (BVF) was (0.32 ± 0.03) in group 1 compared to group 2 (0.24 ± 0.05). Bone mineral density (BMD) was (1.11 ± 0.03) in group 1 compared to group 2 (1.01 ± 0.02). Surgical modifications provide a reliable way for establishment of the murine calvaria osteolysis model