Constructing Linear-Oriented Pre-Vascularized Human Spinal Cord Tissues for Spinal Cord Injury Repair.

Repairing spinal cord injury (SCI) is a global medical challenge lacking effective clinical treatment. Developing human-engineered spinal cord tissues that can replenish lost cells and restore a regenerative microenvironment offers promising potential for SCI therapy. However, creating vascularized human spinal cord-like tissues (VSCT) that mimic the diverse cell types and longitudinal parallel structural features of spinal cord tissues remains a significant hurdle. In the present study, VSCTs are engineered using embryonic human spinal cord-derived neural and endothelial cells on linear-ordered collagen scaffolds (LOCS). Studies have shown that astrocytes and endothelial cells align along the scaffolds in VSCT, supporting axon extension from various human neurons myelinated by oligodendrocytes. After transplantation into SCI rats, VSCT survives at the injury sites and promotes endogenous neural regeneration and vascularization, ultimately reducing scarring and enhancing behavioral functional recovery. It suggests that pre-vascularization of engineered spinal cord tissues is beneficial for SCI treatment and highlights the important role of exogenous endothelial cells in tissue engineering.

Hedgehog pathway negatively regulated depleted uranium-induced nephrotoxicity.

Depleted uranium (DU) retains the radiological toxicities, which accumulates preferentially in the kidneys. Hedgehog (Hh) pathway plays a critical role in tissue injury. However, the role of Hh in DU-induced nephrotoxicity was still unclear. This study was carried out to investigate the effect of Gli2, which was an important transcription effector of Hh signaling, on DU induced nephrotoxicity. To clarify it, CK19 positive tubular epithelial cells specific Gli2 conditional knockout (KO) mice model was exposed to DU, and then histopathological damage and Hh signaling pathway activation was analyzed. Moreover, HEK-293 T cells were exposed to DU with Gant61 or Gli2 overexpression, and cytotoxicity of DU as analyzed. Results showed that DU caused nephrotoxicity accompanied by activation of Hh signaling pathway. Meanwhile, genetic KO of Gli2 reduced DU-induced nephrotoxicity by normalizing biochemical indicators and reducing Hh pathway activation. Pharmacologic inhibition of Gli1/2 by Gant61 reduced DU induced cytotoxicity by inhibiting apoptosis, ROS formation and Hh pathway activation. However, overexpression of Gli2 aggravated DU-induced cytotoxicity by increasing the levels of apoptosis and ROS formation. Taken together, these results revealed that Hh signaling negatively regulated DU-inducted nephrotoxicity, and that inhibition of Gli2 might serve as a promising nephroprotective target for DU-induced kidney injury.

Development of a Multi-Parametric ultrasonography nomogram for prediction of invasiveness in ductal carcinoma in situ.

OBJECTIVE: To investigate the independent risk variables associated with the potential invasiveness of ductal carcinoma in situ (DCIS) on multi-parametric ultrasonography, and further construct a nomogram for risk assessment. METHODS: Consecutive patients from January 2017 to December 2022 who were suspected of having ductal carcinoma in situ (DCIS) based on magnetic resonance imaging or mammography were prospectively enrolled. Histopathological findings after surgical resection served as the gold standard. Grayscale ultrasound, Doppler ultrasound, shear wave elastography (SWE), and contrast-enhanced ultrasound (CEUS) examinations were preoperative performed. Binary logistic regression was used for multifactorial analysis to identify independent risk factors from multi-parametric ultrasonography. The correlation between independent risk factors and pathological prognostic markers was analyzed. The predictive efficacy of DCIS associated with invasiveness was assessed by logistic analysis, and a nomogram was established. RESULTS: A total of 250 DCIS lesions were enrolled from 249 patients, comprising 85 pure DCIS and 165 DCIS with invasion (DCIS-IDC), of which 41 exhibited micro-invasion. The multivariate analysis identified independent risk factors for DCIS with invasion on multi-parametric ultrasonography, including image size (>2cm), Doppler ultrasound RI (≥0.72), SWE's Emax (≥66.4 kPa), hyper-enhancement, centripetal enhancement, increased surrounding vessel, and no contrast agent retention on CEUS. These factors correlated with histological grade, Ki-67, and human epidermal growth factor receptor 2 (HER2) (P < 0.1). The multi-parametric ultrasound approach demonstrated good predictive performance (sensitivity 89.7 %, specificity 73.8 %, AUC 0.903), surpassing single US modality or combinations with SWE or CEUS modalities. Utilizing these factors, a predictive nomogram achieved a respectable performance (AUC of 0.889) for predicting DCIS with invasion. Additionally, a separate nomogram for predicting DCIS with micro-invasion, incorporating independent risk factors such as RI (≥0.72), SWE's Emax (≥65.2 kPa), and centripetal enhancement, demonstrated an AUC of 0.867. CONCLUSION: Multi-parametric ultrasonography demonstrates good discriminatory ability in predicting both DCIS with invasion and micro-invasion through the analysis of lesion morphology, stiffness, neovascular architecture, and perfusion. The use of a nomogram based on ultrasonographic images offers an intuitive and effective method for assessing the risk of invasion in DCIS. Although the nomogram is not currently considered a clinically applicable diagnostic tool due to its AUC being below the threshold of 0.9, further research and development are anticipated to yield positive outcomes and enhance its viability for clinical utilization.

Predictive value of contrast-enhanced ultrasonography and ultrasound elastography for management of BI-RADS category 4 nonpalpable breast masses.

PURPOSE: The objective of this study was to investigate the independent risk factors and associated predictive values of contrast-enhanced ultrasound (CEUS), shear wave elastography (SWE), and strain elastography (SE) for high-risk lesions (HRL) and malignant tumors (MT) among nonpalpable breast masses classified as BI-RADS category 4 on conventional ultrasound. METHODS: This prospective study involved consecutively admitted patients with breast tumors from January 2018, aiming to explore the management of BI-RADS category 4 breast tumors using CEUS and elastography. We conducted a retrospective review of patient data, focusing on those with a history of a nonpalpable mass as the primary complaint. Pathologic findings after surgical resection served as the gold standard. The CEUS arterial-phase indices were analyzed using contrast agent arrival-time parametric imaging processing mode, while quantitative and qualitative indices were examined on ES images. Independent risk factors were identified through binary logistic regression multifactorial analysis. The predictive efficacy of different modalities was compared using a receiver operating characteristics curve. Subsequently, a nomogram for predicting the risk of HRL/MT was established based on a multifactorial logistic regression model. RESULTS: A total of 146 breast masses from 146 patients were included, comprising 80 benign tumors, 12 HRLs, and 54 MTs based on the final pathology. There was no significant difference in pathologic size between the benign and HRL/MT groups [8.00(6.25,10.00) vs. 9.00(6.00,10.00), P = 0.506]. The diagnostic efficacy of US plus CEUS exceeded that of US plus SWE/SE for BI-RADS 4 nonpalpable masses, with an AUC of 0.954 compared to 0.798/0.741 (P ï¼œ 0.001). Further stratified analysis revealed a more pronounced improvement for reclassification of BI-RADS 4a masses (AUC: 0.943 vs. 0.762/0.675, P ï¼œ 0.001) than BI-RADS 4b (AUC:0.950 vs. 0.885/0.796, P＞0.05) with the assistance of CEUS than SWE/SE. Employing downgrade CEUS strategies resulted in negative predictive values ranging from 95.2 % to 100.0 % for BI-RADS 4a and 4b masses. Conversely, using upgrade nomogram strategies, which included the independent predictive risk factors of irregular enhanced shape, poor defined enhanced margin, earlier enhanced time, increased surrounding vessels, and presence of contrast agent retention, the diagnostic performance achieved an AUC of 0.947 with good calibration. CONCLUSION: After investigating the potential of CEUS and ES in improving risk assessment and diagnostic accuracy for nonpalpable BI-RADS category 4 breast masses, it is evident that CEUS has a more significant impact on enhancing classification compared to ES, particularly for BI-RADS 4a subgroup masses. This finding suggests that CEUS may offer greater benefits in improving risk assessment and diagnostic accuracy for this specific subgroup of breast masses.

Predicting axillary metastasis in breast cancer using lymphatic contrast-enhanced ultrasound-guided fine-needle aspiration of one lymph node.

OBJECTIVES: Fine-needle aspiration (FNA) is a microinvasive method to diagnose lymph nodes. This study aims to determine the capability of lymphatic contrast-enhanced ultrasound (LCEUS)-guided FNA in predicting the axillary metastasis with the target of one lymph node (LN) in patients with breast cancer. METHODS: LCEUS was prospectively performed in 105 patients with breast cancer. The most suspicious LN was targeted based on the characters of LCEUS. FNA was performed in the LN, followed by localization using a guide wire. The detection of lymph cells and/or tumour cells was recognized as a puncture success. Cytologic diagnosis was compared with histologic diagnosis of wire-marked LN for diagnosing accuracy and compared with histologic diagnosis of axillary LNs for predicting accuracy. RESULTS: LCEUS-guided FNA was performed in all 105 female patients who underwent axillary dissection. The puncture success rates were 74.3%, 91.4%, and 97.1% for three sequential groups (P = .010). In diagnosing LN metastasis, the sensitivity, specificity, and accuracy values of LCEUS-guided FNA were 89.7%, 100%, and 95.7%, respectively. In predicting axillary metastasis, the sensitivity, specificity, and accuracy values of LCEUS-guided FNA were 81.4%, 100%, and 91.3%, respectively. CONCLUSIONS: The microinvasive LCEUS-guided FNA of one lymph node can be an accurate method and may help predict axillary metastasis in patients with breast cancer. ADVANCES IN KNOWLEDGE: This study presented that LCEUS combined with FNA would be practical in clinic. The characters of LCEUS could indicate the suspicious LNs and promote the accuracy in predicting axillary metastasis.

The relationships between screen exposure, parent-child interactions and comprehension in 8-month-old infants: The mediating role of shared viewing and parent-child conversation.

OBJECTIVE: To explore the relationships between screen exposure, parent-child interactions and comprehension in 8-month-old infants, and to examine whether shared viewing and parent-child conversation during screen exposure may play mediating role in that relationships. METHODS: The sample included 437 infants aged 8 months from the Children's Health Department of Guiyang Maternal and Child Health Hospital during January 2022 to February 2023. The use of electronic screen devices was assessed using a screen exposure questionnaire. The Brigance Parent-child interactions Scale was used to assess parent-child interactions and the Putonghua Communicative Development Inventory (PCDI) scale was used to assess infants' word comprehension. RESULTS: 48.7% of infants were found to be using screens 1-2 days per week. There was a significant difference (p < 0.05) in the PCDI-comprehension scores of screen-exposed infants compared to non-screen-exposed infants. Shared viewing and parent-child conversation during screen exposure were positively associated with parent-child interactions (p < 0.05). Mediation analysis revealed that parent-child conversation fully mediated between screen exposure and PCDI-comprehension, but partially mediated between parent-child interactions and PCDI-comprehension. CONCLUSIONS: Shared viewing and parent-child conversation during screen exposure may mediate between screen exposure and comprehension development. Shared viewing, parent-child conversation and parent-child interactions may be protective factors for screen exposure in comprehension development. Suggests that parents should accompany and communicate with their children when they use electronic screen devices to reduce the negative impact of screen exposure on children's comprehension.

Comparison of the diagnostic value of contrast-enhanced ultrasound combined with conventional ultrasound versus magnetic resonance imaging in malignant non-mass breast lesions.

OBJECTIVE: To compare the diagnostic value of contrast-enhanced ultrasound (CEUS)+conventional ultrasound vs MRI for malignant non-mass breast lesions (NMLs). METHODS: A total of 109 NMLs detected by conventional ultrasound and examined by both CEUS and MRI were retrospectively analysed. The characteristics of NMLs in CEUS and MRI were noted, and agreement between the two modalities was analysed. Sensitivity, specificity, positive-predictive value (PPV), negative-predictive value (NPV), and area under the curve (AUC) of the two methods for diagnosing malignant NMLs were calculated in the overall sample and subgroups of different sizes(<10 mm, 10-20 mm, >20 mm). RESULTS: A total of 66 NMLs detected by conventional ultrasound showed non-mass enhancement in MRI. Agreement between ultrasound and MRI was 60.6%. Probability of malignancy was higher when there was agreement between the two modalities. In the overall group, the sensitivity, specificity, PPV, and NPV of the two methods were 91.3%, 71.4%, 60%, 93.4% and 100%, 50.4%, 59.7%, 100%, respectively. The diagnostic performance of CEUS+conventional ultrasound was better than that of MRI (AUC: 0.825 vs 0.762, p = 0.043). The specificity of both methods decreased as lesion size increased, but sensitivity did not change. There was no significant difference between the AUCs of the two methods in the size subgroups (p > 0.05). CONCLUSION: The diagnostic performance of CEUS+conventional ultrasound may be better than that of MRI for NMLs detected by conventional ultrasound. However, the specificity of both methods decrease significantly as lesion size increases. ADVANCES IN KNOWLEDGE: This is the first study to compare the diagnostic performance of CEUS+conventional ultrasound vs that of MRI for malignant NMLs detected by conventional ultrasound. While CEUS+conventional ultrasound appears to be superior to MRI, subgroup analysis suggests that diagnostic performance is poorer for larger NMLs.

Intratumor microbiota: Occult participants in the microenvironment of multiple myeloma.

More recently, microbiota was detected in several tumorous tissues including multiple myeloma (MM), but the roles of which is still under-studied as paucity of research on tumor biology. Moreover, we also detected the presence of microbiota in the bone marrow of patients with MM by 2bRAD-M sequencing technology, which is an incurable hematological malignancy characterized by accumulation of abnormal plasma cells in the bone marrow. However, the roles of intratumor microbiota in tumor disease remains poorly understood. In this review, we critically reviewed recent literature about microbiota in the tumorigenesis and progression of MM. Importantly, we proposed that the emergence of microbiota in the microenvironment of multiple myeloma may be attributed to microbial dysbiosis and impaired intestinal barrier, due to the increased prevalence of MM in patients with obesity and diabetes, of which the characteristic phenotype is gut microbial dysbiosis and impaired intestinal barrier. When the intestinal barrier is damaged, dysbiotic microbiota and their metabolites, as well as dysregulated immune cells, may participate in the reshaping of the local immune microenvironment, and play pivotal roles in the tumorigenesis and development of multiple myeloma, probably by migrating to the bone marrow microenvironment from intestine. We also discuss the emerging microbiological manipulation strategies to improve long-term outcomes of MM, as well as the prospective of the state-of-the-art techniques to advance our knowledge about the biological implication in the microbiome in MM.

Value of an expanded range of lesions on contrast-enhanced ultrasound for the diagnosis of hypervascular breast masses.

Background: Breast cancer lesions show an expanded range on contrast-enhanced ultrasound (CEUS). Here, we quantitatively analyze this index to explore its effective cutoff value for distinguishing benign and malignant lesions and the corresponding diagnostic performance and investigate its role in prognostic assessment of malignant lesions. Methods: Consecutive patients who underwent CEUS for breast lesions during the period from September 2017 to June 2019 were included. Original CEUS images were selected, displayed in dual-frame mode, and measured when enhancement of the lesion reached its peak. The longitudinal diameter, transverse diameter, and area of the lesion on the two-dimensional images and the corresponding postenhancement images were measured to calculate six indicators: longitudinal diameter increment, transverse diameter increment, area increment, percent increase in longitudinal diameter, percent increase in transverse diameter, and percent increase in area increment. With postoperative pathology as the gold standard, the cutoff values for distinguishing benign and malignant lesions and the correlations of these indicators with pathological subtypes and pathological grades were evaluated. Results: Malignant lesions showed a more significantly expanded range after enhancement compared to benign lesions, especially in terms of area increase. When the cutoff value of the area increment was set at 0.47 cm2 for distinguishing between benign and malignant lesions, the area under the curve (AUC) was 0.945, and the sensitivity, specificity, accuracy, positive predictive value, and negative predictive value were 90.1%, 91.5%, 90.9%, 87.2%, and 93.5%, respectively. The pathologically measured maximum diameter of malignant masses correlated with the percent increase in transverse diameter, area increment, and percent increase in area increment. The longitudinal diameter increment in the luminal A group was significantly smaller than that in the human epidermal growth factor receptor 2 (HER2)+ group. The percent increase in transverse diameter was helpful for predicting the pathological grade of malignant masses. When the cutoff value of the percent increase in transverse diameter was set at 10.84% for pathological grading, the AUC was 0.623, and the sensitivity was 90.8%. Conclusions: Indicators related to the expanded lesion range on CEUS are helpful in differential diagnosis of benign and malignant lesions and in prognostic assessment of pathological grades.

Advances in biomaterial-based cardiac organoids.

Cardiovascular disease (CVD) is one of the important causes of death worldwide. The incidence and mortality rates are increasing annually with the intensification of social aging. The efficacy of drug therapy is limited in individuals suffering from severe heart failure due to the inability of myocardial cells to undergo regeneration and the challenging nature of cardiac tissue repair following injury. Consequently, surgical transplantation stands as the most efficient approach for treatment. Nevertheless, the shortage of donors and the considerable number of heart failure patients worldwide, estimated at 26 million, results in an alarming treatment deficit, with only around 5000 heart transplants feasible annually. The existing major alternatives, such as mechanical or xenogeneic hearts, have significant flaws, such as high cost and rejection, and are challenging to implement for large-scale, long-term use. An organoid is a three-dimensional (3D) cell tissue that mimics the characteristics of an organ. The critical application has been rated in annual biotechnology by authoritative journals, such as Science and Cell. Related industries have achieved rapid growth in recent years. Based on this technology, cardiac organoids are expected to pave the way for viable heart repair and treatment and play an essential role in pathological research, drug screening, and other areas. This review centers on the examination of biomaterials employed in cardiac repair, strategies employed for the reconstruction of cardiac structure and function, clinical investigations pertaining to cardiac repair, and the prospective applications of cardiac organoids. From basic research to clinical practice, the current status, latest progress, challenges, and prospects of biomaterial-based cardiac repair are summarized and discussed, providing a reference for future exploration and development of cardiac regeneration strategies.

Role of NF-κB signaling pathway in hexavalent chromium-induced hepatotoxicity.

Hexavalent chromium Cr (VI) is a primary human carcinogen with damaging toxic effects on multiple organs. Cr (VI) exposure can induce hepatotoxicity through oxidative stress, but its exact mechanism of action was still unclear. In our study, a model of acute Cr (VI) induced liver injury was established by exposing mice to different concentrations (0, 40, 80, and 160 mg/kg) of Cr (VI); RNA-seq was used to characterize changes in liver tissue transcriptome of C57BL/6 mice after exposing to 160 mg/kg Bw of Cr (VI). Changes in liver tissue structures, proteins, and genes were observed by hematoxylin and eosin (H&E), western blot, immunohistochemistry and RT-PCR. After Cr (VI) exposure, abnormal liver tissue structure, hepatocyte injury, and hepatic inflammatory response were observed in mice in a dose-dependent manner. RNA-seq transcriptome results indicated that oxidative stress, apoptosis, and inflammatory response pathways were increased after Cr (VI) exposure; KEGG pathway analysis found that activation of NF-κB signaling pathway was significantly upregulated. Consistent with the RNA-seq results, immunohistochemistry showed that Cr (VI) exposure resulted in infiltrating of Kupffer cells and neutrophils, increasing expression of inflammatory factors (TNF-α, IL-6, IL-1ß), and activating of NF-κB signaling pathways (p-IKKα/ß and p-p65). However, ROS inhibitor, N-acetyl-L-cysteine (NAC), could reduce infiltration of Kupffer cells and neutrophils and expression of inflammatory factors. Besides, NAC could inhibit NF-κB signaling pathway activation, and alleviate Cr (VI)-induced liver tissue damage. Our findings strongly suggested that inhibition of ROS by NAC might help in the development of new strategies for Cr (VI)-associated liver fibrosis. Our findings revealed for the first time that Cr (VI) induced liver tissue damage through the inflammatory response mediated by the NF-κB signaling pathway, and inhibition of ROS by NAC might help in the development of new strategies for Cr (VI)-associated hepatotoxicity.

N-acetyl-L-cysteine attenuated the toxicity of ZIF-8 on EA.hy926 endothelial cells by wnt/ß-catenin pathway.

As kinds of porous crystalline compounds, zeolitic imidazolate frameworks (ZIFs) have been developed quickly and attracted considerable attention for use in nano drug delivery systems, which raised concerns about cardiovascular disorders. At the present, the cytotoxic mechanism of ZIFs in cardiovascular disorders was still unclear. Our experiment explored the toxicity of ZIF-8, a typical kind of ZIFs, on human EA.hy926 vascular endothelial cells. The cell viability, ROS formation, apoptosis level, inflammatory response level, wound healing ability and atherosclerosis-related indicators of EA.hy926 endothelial cells were analyzed after ZIF-8 treatment. Meanwhile, we evaluated the ability of antioxidant N-Acetyl-L-cysteine (NAC) to attenuate the toxicity of ZIF-8 on EA.hy926 endothelial cells. As results, NAC attenuated ROS formation, cell apoptosis, LDH formation and endothelial dysfunction caused by ZIF-8. As the Wnt/ß-catenin pathway was involved in endothelial cell dysfunction, we also studied the expression level of ß-catenin and LEF1 in ZIF-8 and/or NAC treated EA.hy926 cells. As expected, ZIF-8 increased the protein expressions of ß-catenin and LEF1in the IC50 group, which was significantly inhibited by co-treatment with NAC. Taken together, this study could help improve our understanding about the mechanism of ZIF-8-induced endothelial cells injury and NAC had therapeutic potential in preventing ZIF-8-associated endothelial dysfunction by wnt/ß-catenin pathway.

Prognostic significance of CD56 antigen in newly diagnosed multiple myeloma: A real-world retrospective study.

The prognostic value of plasma cell CD56 expression of patients with multiple myeloma (MM) has been reported in many studies, but the results are controversial. This study aimed to examine the prognostic significance of CD56 in MM patients. Eighty seven patients with newly diagnosed MM were enrolled in this study, and their clinical characteristics, immunophenotypes, and cytogenetics were retrospectively analyzed to explore the prognostic significance of CD56 expression. Multiparameter flow cytometry was used to detect MM in bone marrow samples from all patients. Patients were divided into 2 groups based on whether they expressed CD56: CD56 + group and CD56 - group. After 4 cycles of chemotherapy, the overall response rate of the CD56 - patients was lower than that of the CD56 + patients (60.0% vs 81.1%, P = .036). Survival analysis showed that the median progression-free survival (PFS) was 10 months for the CD56 - group and 27 months for the CD56 + group (P = .007). The median overall survival (OS) of patients for the CD56 - group was 25 months versus not reached in the CD56 + group (P = .010). In addition, among the high-risk patients detected by fluorescence in situ hybridization (FISH), the median PFS was 4 months for the CD56 - group and 16 months for the CD56 + group (P = .012). The median OS of the CD56 + group and CD56 - group was 36 months and 15 months, respectively, with statistically significant differences (P = .017). Our study confirmed that CD56 - patients with MM had a worse prognosis than that of CD56 + patients with MM. Among the patients with ≥ 2 high-risk cytogenetics, the existence of the CD56 negativity can further identify MM patients with poor PFS and OS.

Endosperm-specific accumulation of human α-lactalbumin increases seed lysine content in maize.

KEY MESSAGE: This study demonstrated high expression and accumulation of human α-lactalbumin in transgenic maize, and significant improvement of lysine content in maize endosperm. As a high-yield crop, lack of lysine in endosperm storage protein is a major defect of maize (Zea mays L.). Specifically expression of foreign proteins is a potential way to improve lysine content in maize endosperm. Human α-lactalbumin is such a protein with high lysine content and high nutritional value. In this study, the codon-optimized human lactalbumin alpha (LALBA) gene was driven by maize endosperm-specific 27 kD γ-zein promoter, and transformed into maize. Five independent transgenic lines were obtained, and LALBA was highly expressed in endosperm in all these lines. Protein assay indicated that human α-lactalbumin was highly accumulated in maize endosperm. Immuno-localization assay indicated that human α-lactalbumin was mainly deposited into the protein body (PB). Protein interaction assay showed that human α-lactalbumin interacted with 16 kD γ-zein, which might lead to its deposition to the PBs. Amino acid analysis of two independent transgenic lines showed significant increase of lysine contents in transgenic endosperm, with 47.26% and 45.15% increase to their non-transgenic seeds, respectively. We obtained transgenic maize with endosperm-specific accumulation of human α-lactalbumin at high level and increased the lysine content in maize endosperm. This study demonstrated an effective way to improve the nutritional value of maize seeds.

miR-576-5p Facilitates Aggressive Cell Behaviors in Colon Adenocarcinoma via Targeting NEGR1.

The microRNA (miRNA) miR-576-5p was reported to facilitate tumor progression, but its underlying regulatory impacts on colon adenocarcinoma remain unknown. This work therefore attempted to examine the biological effects of miR-576-5p in colon adenocarcinoma. The Cancer Genome Atlas (TCGA) database was introduced to analyze miR-576-5p and NEGR1 messenger RNA (mRNA) expression levels between normal and cancer tissues. miR-576-5p and NEGR1 expression levels in colon adenocarcinoma cells and colon cells were evaluated with quantitative reverse transcription polymerase chain reaction (qRT-PCR). NEGR1 protein expression was examined by Western blot. Furthermore, colon adenocarcinoma cell behaviors were evaluated via CCK-8, wound-healing, Transwell, and hanging drop experiments. The interaction between miR-576-5p and NEGRI was verified by dual-luciferase assay. miR-576-5p was upregulated in colon adenocarcinoma, and miR-576-5p overexpression notably facilitated the proliferative, migratory, invasive abilities of colon adenocarcinoma cells. NEGR1 was newly identified as one target of miR-576-5p, and, miR-576-5p/NEGR1 axis was subsequently verified to modulate cell proliferative, migratory, invasive, and aggregate abilities. miR-576-5p facilitated aggressive progression of colon adenocarcinoma cells by targeting NEGR1, which could be an underlying therapeutic target for colon adenocarcinoma.

Proteomics of post-translational modifications in colorectal cancer: Discovery of new biomarkers.

Colorectal cancer (CRC) is one of the costliest health problems and ranks second in cancer-related mortality in developed countries. With the aid of proteomics, many protein biomarkers for the diagnosis, prognosis, and precise management of CRC have been identified. Furthermore, some protein biomarkers exhibit structural diversity after modifications. Post-translational modifications (PTMs), most of which are catalyzed by a variety of enzymes, extensively increase protein diversity and are involved in many complex and dynamic cellular processes through the regulation of protein function. Accumulating evidence suggests that abnormal PTM events are associated with a variety of human diseases, such as CRC, thus highlighting the need for studying PTMs to discover both the molecular mechanisms and therapeutic targets of CRC. In this review, we begin with a brief overview of the importance of protein PTMs, discuss the general strategies for proteomic profiling of several key PTMs (including phosphorylation, acetylation, glycosylation, ubiquitination, methylation, and citrullination), shift the emphasis to describing the specific methods used for delineating the global landscapes of each of these PTMs, and summarize the recent applications of these methods to explore the potential roles of the PTMs in CRC. Finally, we discuss the current status of PTM research on CRC and provide future perspectives on how PTM regulation can play an essential role in translational medicine for early diagnosis, prognosis stratification, and therapeutic intervention in CRC.

The Positive Influence of Polyphenols Extracted From Pueraria lobata Root on the Gut Microbiota and Its Antioxidant Capability.

Pueraria lobata, an edible food and medicinal plant, is a rich source of bioactive components. In this study, a polyphenol-rich extract was isolated from P. lobata. Puerarin was identified, and the high antioxidant bioactivity of the P. lobata extract was evaluated using the methods of 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS), and hydroxyl free radical scavenging ratio. Additionally, the IC50 values of DPPH, ABTS, and hydroxyl radical scavenging activities were 50.8, 13.9, and 100.4 µg/ml, respectively. Then, the P. lobata extract was administered to C57Bl/6J mice and confirmed to have a superior effect on enhancing the antioxidant status including improving superoxide dismutase activity, glutathione peroxidase peroxide activity, total antioxidant capacity activity, and malondialdehyde contents in vivo. Furthermore, the P. lobata extract had beneficial and prebiotic effects on the composition and structure of gut microbiota. Results showed that the P. lobata extract significantly increased the abundance of beneficial bacteria, involving Lactobacillaceae and Bacteroidetes, and decreased the abundance of Ruminococcaceae, Prevotellaceae, and Burkholderiaceae. Overall, our results provided a basis for using the P. lobata extract as a promising and potential functional ingredient for the food industry.

[Relationship between the Expression of miRNA181a-5p and the Imbalance of Treg/Th17 in Primary Immune Thrombocytopenia].

OBJECTIVE: To investigate the role of relationship between the expression of miRNA181a-5p and imbalance of Treg/Th17 in the pathogenesis of primary immune thrombocytopenia(ITP), which contributes to clarify the mechanism of T cell immune imbalance in ITP patients. METHODS: Peripheral blood was collected from 37 ITP patients, concluding 21 untreated patients and 16 effectively treated patients, and 19 healthy controls; Peripheral blood mononuclear cells (PBMC) were isolated and the expression of miRNA181a-5p and Notch1 was analyzed by RT-PCR. The proportion of Th17 subsets and Treg cells in the peripheral circulation was detected by flow cytometer (FCM). Clinical data of ITP group was collected, including age, platelet count and disease course. RESULTS: The expression of miR-181a-5p was significantly decreased in ITP group than that of healthy control group (P＜0.01). After effective treatment, the expression of miR-181a-5p was significantly higher than that of ITP group (P＜0.05), but still significantly lower than that of healthy control group (P＜0.01); The expression of Notch1 was significantly increased in ITP group and effectively treated group than that of healthy control group (P＜0.01). There was no significant difference in proportion of Treg cells in ITP group, effectively treated group and healthy control group (P＞0.05). The proportion of Th17 subsets in ITP group was significantly increased than that of healthy control group (P＜0.05), while the ratio of Treg/Th17 was significantly decreased (P＜0.05). There was a positive correlation between the expression of miR-181a-5p and ratio of Treg/Th17 in ITP group (r=0.555). CONCLUSION: The expression of miR-181a-5p is significantly decreased in ITP patients, which is closely related to the imbalance of Treg/Th17 cells. After effective treatment, the expression of miR-181a-5p can be significantly corrected, but still failed to reach the level of healthy people. While the expression of Notch1 is significantly increased in ITP patients, and could not reach the level of healthy people after effective treatment.

Diagnostic value of Doppler imaging for malignant non-mass breast lesions: with different diagnostic criteria for older and younger women: first results.

OBJECTIVE: To evaluate and optimize the additional diagnostic value of Doppler imaging for malignant NMLs detected by US. MATERIALS AND METHODS: The characteristics of 233 NMLs in Doppler imaging were analyzed, and different Adler grades of intralesional vessels were selected as the diagnostic cutoffs on Doppler imaging: grade 1 in the full cohort and in women < 40 years, and grade 0 in women ≥40 years. The diagnostic performance of US and US + Doppler imaging were calculated and compared with that of mammography. RESULTS: The AUC of US + Doppler was larger than that of US alone in each group (P < 0.001). In the full cohort, addition of Doppler imaging increased specificity of US, but decreased sensitivity. However, by use of different diagnostic cutoffs in the two subgroups, it was possible to achieve high sensitivity and specificity simultaneously, which were 100% and 75.8% in women < 40 years, 94.7% and 69.5% in women ≥40 years, respectively. The AUC + Doppler was comparable to that of mammography in the full cohort and in women ≥40 years. In women < 40 years, the AUC of the combination was larger than that of mammography (P < 0.001). CONCLUSION: Doppler imaging, with different Adler grades used as cutoffs in older versus younger women, can improve the specificity of US for the diagnosis of malignant NMLs without losing sensitivity. In younger women, US + Doppler imaging may be better than mammography.

Hexavalent chromium induces hepatocyte apoptosis via regulation of apoptosis signal-regulating kinase 1/c-Jun amino-terminal kinase signaling.

With the spread of hexavalent chromium (Cr(VI)) contamination, Cr(VI)-induced hepatotoxicity has attracted increasing attention in recent years. To date, however, the exact mechanism of Cr(VI) toxicity remains unclear. In this study, we investigated the role of apoptosis signal-regulating kinase 1 (ASK1)/c-Jun amino-terminal kinase (JNK) in Cr(VI)-induced hepatic toxicity and the possible related mechanisms. AML-12 hepatocyte cell-lines were treated with 0, 1, 4, and 16 µmol/Lof Cr(VI) with or without GS-444271 (an ASK1 inhibitor). Adult male mice were administered with 0, 2, 8, and 32 mg/kg body mass (BM)/day of Cr(VI) for 5 days. The level of hepatocyte apoptosis/proliferation, generation of reactive oxygen species (ROS), and expression levels of mRNAs and proteins related to ASK1/JNK and nuclear factor-E2-related factor 2 (Nrf2) signaling were assessed. Results showed that high Cr(VI) exposure induced hepatocyte apoptosis and liver injury by generation of ROS and down-regulation of Nrf2 signaling. In addition, ASK1/JNK signaling activity was upregulated in the Cr(VI)-treated group. Furthermore, GS-444217 treatment significantly rescued Cr(VI)-induced hepatocyte apoptosis and liver dysfunction in vitro and in vivo by down-regulation of ASK1/JNK signaling. Thus, ASK1/JNK signaling appears to play an important role in Cr(VI)-induced hepatocyte apoptosis and liver injury. This study should help improve our understanding of the mechanism of Cr(VI)-induced liver injury and provide support for future investigations on liver disease therapy.