Breast cancer stem cells are a group of undifferentiated cells with self-renewal and multidifferentiation potential. Chemotherapeutic and radiotherapeutic resistance, hypoxic resistance, high tumorigenicity, high cell invasion, and metastatic abilities are characteristics of these cells, which are responsible for breast cancer recurrence. Therefore, the correct sorting and identification of breast cancer stem cells is a primary step for research in this field. This article briefly describes the recent progress on sorting and identification technologies for breast cancer stem cells. Sorting technologies include the side population technique, technologies that depend on cell surface markers, ALDEFLUOR assays, and in situ detection. Identification technologies include mammosphere cultures, limited dilution in vitro, and in-vivo animal models. This review provides an important reference for breast cancer stem cell research, which will explore new methods for the treatment of patients with breast cancer.
AC133 Antigen ; ATP Binding Cassette Transporter, Sub-Family G, Member 2 ; ATP-Binding Cassette Transporters ; metabolism ; Aldehyde Dehydrogenase ; metabolism ; Animals ; Antigens, CD ; metabolism ; Breast Neoplasms ; pathology ; Female ; Flow Cytometry ; methods ; Glycoproteins ; metabolism ; Humans ; Hyaluronan Receptors ; metabolism ; Integrin alpha6 ; metabolism ; Integrin beta1 ; metabolism ; Integrin beta3 ; metabolism ; Isoenzymes ; metabolism ; Membrane Proteins ; metabolism ; Neoplasm Proteins ; metabolism ; Neoplastic Stem Cells ; metabolism ; pathology ; Peptides ; metabolism ; Retinal Dehydrogenase ; metabolism ; Side-Population Cells ; cytology ; metabolism

OBJECTIVETo express and purify recombinant human interleukin-35[IL-35-IgG1 (Fc) in eukaryotic expression system and to study the interaction of IL-35 with gp130 protein.

METHODSA mammalian expression vector, pSTEP2-IL35-LFc, was constructed and transfected into HEK293T cells. Then rhIL-35-IgG1 (Fc) was expressed and purified with protein A affinity chromatography, and was examined with SDS-PAGE and Western blot analysis. The binding of IL-35 to its receptor gp130 was investigated using enzyme-linked immunosorbent assay (ELISA). The biological effect of IL-35 on gp130 was explored in M1 myeloid leukemia cells.

RESULTSrhIL-35-Fc with high purity on reduced SDS-PAGE was obtained. ELISA confirmed that IL-35-Fc was bound to gp130 and neutralized the function of gp130 in M1 myeloid leukemic cells.

CONCLUSIONSHigh-purity and biologically active rhIL-35-Fc protein successfully produced in this study. IL-35 binds to gp130 and neutralizes its activity of in M1 myeloid leukemic cells.

Keloids are benign skin tumors resulting from the excessive proliferation of connective tissue in wound skin. Precise prediction of keloid risk in trauma patients and timely early diagnosis are of paramount importance for in-depth keloid management and control of its progression. This study analyzed four keloid datasets in the high-throughput gene expression omnibus (GEO) database, identified diagnostic markers for keloids, and established a nomogram prediction model. Initially, 37 core protein-encoding genes were selected through weighted gene co-expression network analysis (WGCNA), differential expression analysis, and the centrality algorithm of the protein-protein interaction network. Subsequently, two machine learning algorithms including the least absolute shrinkage and selection operator (LASSO) and the support vector machine-recursive feature elimination (SVM-RFE) were used to further screen out four diagnostic markers with the highest predictive power for keloids, which included hepatocyte growth factor (HGF), syndecan-4 (SDC4), ectonucleotide pyrophosphatase/phosphodiesterase 2 (ENPP2), and Rho family guanosine triphophatase 3 (RND3). Potential biological pathways involved were explored through gene set enrichment analysis (GSEA) of single-gene. Finally, univariate and multivariate logistic regression analyses of diagnostic markers were performed, and a nomogram prediction model was constructed. Internal and external validations revealed that the calibration curve of this model closely approximates the ideal curve, the decision curve is superior to other strategies, and the area under the receiver operating characteristic curve is higher than the control model (with optimal cutoff value of 0.588). This indicates that the model possesses high calibration, clinical benefit rate, and predictive power, and is promising to provide effective early means for clinical diagnosis.
Humans ; Keloid/genetics* ; Nomograms ; Algorithms ; Calibration ; Machine Learning

Precise segmentation of lung field is a crucial step in chest radiographic computer-aided diagnosis system. With the development of deep learning, fully convolutional network based models for lung field segmentation have achieved great effect but are poor at accurate identification of the boundary and preserving lung field consistency. To solve this problem, this paper proposed a lung segmentation algorithm based on non-local attention and multi-task learning. Firstly, an encoder-decoder convolutional network based on residual connection was used to extract multi-scale context and predict the boundary of lung. Secondly, a non-local attention mechanism to capture the long-range dependencies between pixels in the boundary regions and global context was proposed to enrich feature of inconsistent region. Thirdly, a multi-task learning to predict lung field based on the enriched feature was conducted. Finally, experiments to evaluate this algorithm were performed on JSRT and Montgomery dataset. The maximum improvement of Dice coefficient and accuracy were 1.99% and 2.27%, respectively, comparing with other representative algorithms. Results show that by enhancing the attention of boundary, this algorithm can improve the accuracy and reduce false segmentation.
X-Rays ; Algorithms ; Diagnosis, Computer-Assisted ; Thorax/diagnostic imaging* ; Lung/diagnostic imaging* ; Image Processing, Computer-Assisted

Bioactive glass (BG) has been widely used in the preparation of artificial bone scaffolds due to its excellent biological properties and non-cytotoxicity, which can promote bone and soft tissue regeneration. However, due to the brittleness, poor mechanical strength, easy agglomeration and uncontrollable structure of glass material, its application in various fields is limited. In this regard, most current researches mainly focus on mixing BG with organic or inorganic materials by freeze-drying method, sol-gel method, etc., to improve its mechanical properties and brittleness, so as to increase its clinical application and expand its application field. This review introduces the combination of BG with natural organic materials, metallic materials and non-metallic materials, and demonstrates the latest technology and future prospects of BG composite materials through the development of scaffolds, injectable fillers, membranes, hydrogels and coatings. The previous studies show that the addition of BG improves the mechanical properties, biological activity and regeneration potential of the composites, and broadens the application of BG in the field of bone tissue engineering. By reviewing the recent BG researches on bone regeneration, the research potential of new materials is demonstrated, in order to provide a reference for future related research.
Bone Regeneration ; Bone and Bones ; Freeze Drying ; Glass ; Hydrogels

OBJECTIVE: To summarize the progress of clinical diagnosis and treatment of diabetic Charcot neuroarthropathy (CNO) of foot and ankle to provide reference for clinical treatment. METHODS: The research literature on diabetic CNO of foot and ankle at home and abroad was widely reviewed, and the stages and classification criteria of CNO were summarized, and the treatment methods at different stages of the disease course were summarized. RESULTS: CNO is a rapidly destructive disease of bone and joint caused by peripheral neuropathy, which leads to the formation of local deformities and stress ulcers due to bone and joint destruction and protective sensory loss, which eventually leads to disability and even life-threatening. At present, the modified Eichenholtz stage is a commonly used staging criteria for CNO of foot and ankle, which is divided into 4 stages by clinical and imaging manifestations. The classification mainly adopts the modified Brodsky classification, which is divided into 6 types according to the anatomical structure. The treatment of diabetic CNO of foot and ankle needs to be considered in combination with disease stage, blood glucose, comorbidities, local soft tissue conditions, degree of bone and joint destruction, and whether ulcers and infections are present. Conservative treatment is mainly used in the active phase and surgery in the stable phase. CONCLUSION The formulation of individualized and stepped treatment regimens can help improve the effectiveness of diabetic CNO of foot and ankle. However, there is still a lack of definitive clinical evidence to guide the treatment of active and stable phases, and further research is needed.
Humans ; Ankle ; Ulcer/complications* ; Arthropathy, Neurogenic/therapy* ; Ankle Joint ; Diabetes Mellitus ; Diabetic Foot/therapy*

A xylosidase gene, labeled as BH1068 in genome of Bacillus halodurans C-125, was successfully cloned and overexpressed in Escherichia coli JM109. The purified enzyme was thoroughly characterized and its xylosidase function was unambiguously confirmed. It has maximum activities in neutral condition and is stable over a wide range of pH (4.5-9.0). The enzyme has a broad temperature optimal (35 degrees C-45 degrees C) and is quite stable at temperature up to 45 degrees C. The unique pH and temperature profiles of the enzyme should allow a wide range of xylanolytic operational conditions. With high specific activity of 174 mU/mg protein for its artificial substrate (p-nitrophenyl-beta-xylose) and low xylose inhibition (inhibitor constant Ki = 300 mmol/L), this enzyme is among the most active and high tolerant bacterial xylosidase to xylose inhibition. Its high synergy with commercial xylanase has been demonstrated with beechwood xylan hydrolysis, achieving a hydrolysis yield of 40%. Its neutral pH optimal and high tolerance to product inhibition complements well with its fungal counterparts that are only optimal at acidic pH and susceptible to xylose inhibition. In conclusion, this enzyme has high potential in the saccharification of xylan and xylan-containing polysaccharides.
Amino Acid Sequence ; Bacillus ; classification ; enzymology ; genetics ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Hydrolysis ; Molecular Sequence Data ; Recombinant Proteins ; genetics ; metabolism ; Substrate Specificity ; Xylose ; metabolism ; Xylosidases ; genetics ; metabolism

The study aimed to evaluate the safety and function of poly(lactic-acid-co-ε-caprolactone) (PLCL)/fibrinogen nanofibers (P/F-Ns), and provide theoretical basis for the clinical application. The surface morphology, mechanical properties, the hydrophilicity and the fibrinogen content of P/F-Ns were tested by scanning electron microscope, the material testing machine, the contact angle meter and the microplate reader, respectively. The cell adhesion, proliferation and ligament remodeling genes expression of Hig-82 cells on P/F-Ns were conducted through cell counting kit-8 (CCK-8) and real-time quantitative PCR analyses, respectively. The results showed that with the increase of the fibrinogen content, the pore sizes and hydrophilicity of three P/F-Ns increased, but the mechanical properties decreased. Cell adhesion and proliferation tests showed that P/F-N-2 held the best ability to promote cell adhesion and proliferation. The ligament remodeling genes expressions of Hig-82 cells on P/F-N-1, P/F-N-2 and P/F-N-3 were all up-regulated compared to P/F-N-0 on days 3 and 7. All the three P/F-Ns containing fibrinogen (P/F-N-1, P/F-N-2 and P/F-N-3) had better biocompatibility compared to P/F-N-0, and could be efficiently applied to the reconstruction of anterior cruciate ligament.
Anterior Cruciate Ligament Reconstruction ; Cell Adhesion ; Fibrinogen ; Materials Testing ; Nanofibers

The pulsatile flow experiment can not only evaluate the preclinical safety and risk of prosthetic heart valve (PHV) but also play an important role in the computational model and fluid simulation, providing an experimental basis for the performance optimization of PHV. This paper mainly reviews the development and the latest progress of PHV pulsatile flow experiments and the characteristics of experimental pulse duplicator, and discuss the research direction of pulsatile flow experiments, expecting a further development in this field.

Considering the small differences between different types in the diabetic retinopathy (DR) grading task, a retinopathy grading algorithm based on cross-layer bilinear pooling is proposed. Firstly, the input image is cropped according to the Hough circle transform (HCT), and then the image contrast is improved by the preprocessing method; then the squeeze excitation group residual network (SEResNeXt) is used as the backbone of the model, and a cross-layer bilinear pooling module is introduced for classification. Finally, a random puzzle generator is introduced in the training process for progressive training, and the center loss (CL) and focal loss (FL) methods are used to further improve the effect of the final classification. The quadratic weighted Kappa (QWK) is 90.84% in the Indian Diabetic Retinopathy Image Dataset (IDRiD), and the area under the receiver operating characteristic curve (AUC) in the Messidor-2 dataset (Messidor-2) is 88.54%. Experiments show that the algorithm proposed in this paper has a certain application value in the field of diabetic retina grading.
Humans ; Diabetic Retinopathy/diagnostic imaging* ; Algorithms ; ROC Curve ; Diabetes Mellitus