ROR2 promotes invasion and chemoresistance of triple-negative breast cancer cells by activating PI3K/AKT/mTOR signaling.

Objective: This study aimed to investigate the role of receptor tyrosine kinase-like orphan receptor 2 (ROR2) in triple-negative breast cancer (TNBC). Methods: ROR2 expression in primary TNBC and metastatic TNBC tissues was analyzed by immunohistochemical staining and PCR. ROR2 expression in TNBC cell lines was detected by PCR and Western blot analysis. The migration, invasion and chemosensitivity of TNBC cells with overexpression or knockdown of ROR2 were examined. Results: ROR2 expression was high in metastatic TNBC tissues. ROR2 knockdown suppressed the migration, invasion and chemoresistance of TNBC cells. ROR2 overexpression in MDA-MB-435 cells promoted the migration, invasion, and chemoresistance. Moreover, ROR2 knockdown in HC1599 and MDA-MB-435 adriamycin-resistant cells enhanced chemosensitivity to adriamycin. ROR2 could activate PI3K/AKT/mTOR signaling in TNBC cells. Conclusion: ROR2 is upregulated and promotes metastatic phenotypes of TNBC by activating PI3K/AKT/mTOR signaling.

Distinct immunoreactions after a primary tumor microwave ablation using different heating parameters in a VX2 tumor model.

BACKGROUND: Thermal ablation of solid tumors in situ can activate the immune system and produce a specific immune response against the tumor. Microwave ablation (MWA) with different parameters can ablate tumors with similar sizes and cause different local inflammatory effects. Our aim was to determine the immunological effects induced by different energy modes of MWA for a primary tumor. METHODS: Seventy rabbits with VX2 tumors that were implanted subcutaneously underneath the right second nipple were treated with high-power MWA (40 W for 1 min), low-power MWA (20 W for 2 min), or surgical resection or were left without treatment (control). Survival time was evaluated by log-rank test. On day 14 after ablation, immunohistochemistry and flow cytometry were used to evaluate the T-cell immune responses. In addition, the cytokine patterns were identified by enzyme-linked immunosorbent assay. RESULTS: Tumor eradication was achieved completely in the MWA groups, as proven by nicotinamide adenine dinucleotide diaphorase staining. Compared with the three treatment groups, the control group had a significantly higher number of pulmonary metastases and worse survival; however, no significant difference was observed among the three treatment groups. More intra-tumoral and systemic CD4+ and CD8+ T-cells were induced in the MWA groups than in the control group. Compared with operation, MWA induced more systemic CD4+ T-cells. More intra-tumoral CD4+ and CD8+ T-cells and systemic CD4+ T-cells were induced by high-power MWA than by low-power MWA. Moreover, MWA increased the interleukin 2 (IL2) and IL12 levels and decreased the IL4, IL6, and IL10 levels. Importantly, the serum IL12 level was significantly higher after high-power MWA than after low-power MWA. CONCLUSION: High-power MWA enhanced the type 1 T helper immune response and may be selected for the treatment of solid tumors. Future studies are needed to confirm our results.

Cementless bipolar hemiarthroplasty compared with proximal femoral nail anti-rotation of unstable intertrochanteric fractures in senile patients with osteoporosis: a retrospective study.

BACKGROUND: Implant design for the correction of osteoporotic unstable intertrochanteric fractures in elderly patients is a controversial issue. Our study aims to compare the efficacy of PFNA and cementless bipolar hemiarthroplasty (CBH) in treating osteoporotic unstable intertrochanteric fractures in the elderly. METHODS: We retrospectively assessed 70 elderly patients, aged > 70 years old, with intertrochanteric fractures (AO/OTA 31-A2 fractures) from 2014 to 2019. Among them, 34 patients received PFNA and 36 patients received CBH, accompanied with 2-year follow-ups. Additionally, the efficacy difference between the two implants was compared. RESULTS: Both groups had similar general variables like age, gender, fracture site, degree of osteoporosis, fracture classification, ASA score, basic diseases, preoperative preparation time, anesthesia mode, amount of postoperative blood loss, hospital length of stay, along with postoperative blood transfusions and postoperative complications (P > 0.05). Conversely, significant differences were observed among intraoperative variables (amount of blood loss, amount of blood transfusions, operative time, number of intraoperative fluoroscopy), postoperative variables (weight-bearing time out of bed), and Harris hip function score within 12 months of operation (P < 0.05). CONCLUSIONS: CBH showed no obvious advantage over PFNA in the perioperative period in elderly patients with osteoporotic unstable intertrochanteric fractures. However, the joint replacement allowed for earlier ambulation after the operation and rapid recovery of the hip joint function.

Identification of HIF-1α/VEGFA signaling pathway and transcription factors in Kashin-Beck disease by integrated bioinformatics analysis.

Kashin-Beck disease (KBD) is a chronic and endemic osteoarthropathy. The pathogenesis of KBD has yet to be fully elucidated, although previous studies have shown that its etiology may be associated with low selenium abundance and high exposure to mycotoxins, such as T-2 toxin. In the present study, the Comparative Toxicogenomics Database was used to identify key genes associated with KBD, T-2 toxin and selenium. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were used to identify the biological processes and pathways that key genes may be associated with. By searching the Search Tool for the Retrieval of Interacting Genes database and the Molecular Complex Detection plug-in with Cytoscape, it was possible to construct a KBD-associated protein-protein interaction (PPI) network, and screen the core modules and genes. Western blot analysis was subsequently used to verify the expression levels of hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor A (VEGFA), two components that are associated with the HIF-1 signaling pathway in KBD disease. Via this approach, a total of 301 key genes were identified that were associated with KBD, T-2 toxin and selenium. The results of the GO and KEGG enrichment analyses demonstrated that these key genes were mainly involved in the process of apoptosis. Previous studies have demonstrated that excessive apoptosis of chondrocytes plays a crucial role in the pathophysiology of KBD, and that HIF-1α has an important role in chondrocyte apoptosis; therefore, the present study was focused on the expression level of HIF-1α in KBD. By analyzing the PPI network constructed from the key genes, a total of 10 core genes were obtained that may be associated with KBD. The results of western blotting experiments revealed that, after treating chondrocytes with different concentrations of T-2 toxin, the expression levels of HIF-1α and VEGFA were markedly downregulated. The iRegulon plug-in for Cytoscape was used to predict the transcription factors that may regulate HIF-1α and VEGFA in the HIF-1 signaling pathway. Using this approach, 10 core genes and 15 transcription factors were obtained. These results may help to clarify the pathogenesis of KBD, thereby providing further avenues for the therapeutic treatment of KBD.

A seven-lncRNA signature for predicting Ewing's sarcoma.

BACKGROUND: Long non-coding RNAs (lncRNAs) are a class of non-coding RNAs with unique characteristics. These RNA can regulate cancer cells' survival, proliferation, invasion, metastasis, and angiogenesis and are potential diagnostic and prognostic markers. We identified a seven-lncRNA signature related to the overall survival (OS) of patients with Ewing's sarcoma (EWS). METHODS: We used an expression profile from the Gene Expression Omnibus (GEO) database as a training cohort to screen out the OS-associated lncRNAs in EWS and further established a seven-lncRNA signature using univariate Cox regression, the least absolute shrinkage, and selection operator (LASSO) regression analysis. The prognostic lncRNA signature was validated in an external dataset from the International Cancer Genome Consortium (ICGC) as a validation cohort. RESULTS: We obtained 10 survival-related lncRNAs from the Kaplan-Meier and ROC curve analysis (log-rank test P < 0.05; AUC >0.6). Univariate Cox regression and LASSO regression analyses confirmed seven key lncRNAs and we established a lncRNA signature to predict an EWS prognosis. EWS patients in the training cohort were categorized into a low-risk group or a high-risk group based on their median risk score. The high-risk group's survival time was significantly shorter than the low-risk group's. This seven-lncRNA signature was further confirmed by the validation cohort. The area under the curve (AUC) for this lncRNA signature was up to 0.905 in the training group and 0.697 in the 3-year validation group. The nomogram's calibration curves demonstrated that EWS probability in the two cohorts was consistent between the nomogram prediction and actual observation. CONCLUSION: We screened a seven-lncRNA signature to predict the EWS patients' prognosis. Our findings provide a new reference for the current prognostic evaluation of EWS and new direction for the diagnosis and treatment of EWS.

Combination of atmospheric and room temperature plasma (ARTP) mutagenesis, genome shuffling and dimethyl sulfoxide (DMSO) feeding to improve FK506 production in Streptomyces tsukubaensis.

FK506 is a clinically important macrocyclic polyketide with immunosuppressive activity produced by Streptomyces tsukubaensis. However, the production capacity of the strain is very low. To improve production, atmospheric and room temperature plasma (ARTP) mutagenesis was adopted to get the initial strains used in genome shuffling (GS). After three rounds of GS, S. tsukubaensis R3-C4 was the most productive strain, resulting in a FK506 concentration of 335 µg/mL, 2.6 times than that of the original wild-type strain. Moreover, exogenous DMSO 4% (v/v) addition could induce efflux of FK506 and increased FK506 production by 27.9% to 429 µg/mL. Finally, analyses of the differences in morphology, fermentation characteristics and specific gene expression levels between S. tsukubaensis R3-C4 and the wild-type strain revealed that R3-C4 strain: has hampered spore differentiation, thicker mycelia and more red pigment, which are likely related to the downregulation of bldD and cdgB expression. In addition, the expression levels of fkbO, fkbP, dahp, pccB and prpE all showed up-regulation at diverse degrees compared to the wild-type S. tsukubaensis. Overall, these results show that a combined approach involving classical random mutation and exogenous feeding can be applied to increase FK506 biosynthesis and may be applied also to the improvement of other important secondary metabolites.

Weighted gene correlation network analysis reveals novel biomarkers associated with mesenchymal stromal cell differentiation in early phase.

Osteoporosis is a major public health problem that is associated with high morbidity and mortality, and its prevalence is increasing as the world's population ages. Therefore, understanding the molecular basis of the disease is becoming a high priority. In this regard, studies have shown that an imbalance in adipogenic and osteogenic differentiation of bone marrow mesenchymal stem cells (MSCs) is associated with osteoporosis. In this study, we conducted a Weighted Gene Co-Expression Network Analysis to identify gene modules associated with the differentiation of bone marrow MSCs. Gene Ontology and Kyoto Encyclopedia of Genes and Genome enrichment analysis showed that the most significant module, the brown module, was enriched with genes involved in cell cycle regulation, which is in line with the initial results published using these data. In addition, the Cytoscape platform was used to identify important hub genes and lncRNAs correlated with the gene modules. Furthermore, differential gene expression analysis identified 157 and 40 genes that were upregulated and downregulated, respectively, after 3 h of MSCs differentiation. Interestingly, regulatory network analysis, and comparison of the differentially expressed genes with those in the brown module identified potential novel biomarker genes, including two transcription factors (ZNF740, FOS) and two hub genes (FOXQ1, SGK1), which were further validated for differential expression in another data set of differentiation of MSCs. Finally, Gene Set Enrichment Analysis suggested that the two most important candidate hub genes are involved in regulatory pathways, such as the JAK-STAT and RAS signaling pathways. In summary, we have revealed new molecular mechanisms of MSCs differentiation and identified novel genes that could be used as potential therapeutic targets for the treatment of osteoporosis.

Effects of Rolling-Sliding Mechanical Stimulation on Cartilage Preserved In Vitro.

INTRODUCTION: Mechanical stimulation is important for maintaining cartilage function. We used a loading device to exert rolling-sliding mechanical stimulation on cartilage preserved in vitro to investigate cartilage viability and the involved mechanisms. METHODS: Osteochondral grafts from pig knees were randomly classified into loading and control groups. The loading group cartilage was subjected to cycles of mechanical stimulation with specified frequency/time/pressure combinations every 3 days; Then the DMEM was refreshed, and the cartilage was preserved in vitro. The control group cartilage was preserved in DMEM throughout the process and was changed every 3 days. On days 14 and 28, the chondrocyte survival rate, histology, and Young's modulus of the cartilage were measured. Western blots were performed after 2 h of loading to evaluate the protein expression. RESULTS: The loading group showed a significantly higher chondrocyte survival rate, proteoglycan and type II collagen content, and Young's modulus than did the control group on day 14, but no statistically significant differences were found on day 28. After two hours of the loading, the phosphorylation levels of MEK and ERK1/2 increased, and the expression of caspase-3, cleaved caspase-3 and bax decreased. CONCLUSION: These results suggest that periodic rolling-sliding mechanical stimulation can increase cartilage vitality in 2 weeks; a possible mechanism is that mechanical stimulation activates the MEK/ERK signalling pathway, thus inhibiting apoptotic protein expression. This loading preservation scheme could be used by cartilage tissue banks to improve cartilage preservation in vitro and enhance the quality of cartilage repair.

Fault Identification of Chemical Processes Based on k-NN Variable Contribution and CNN Data Reconstruction Methods.

Data-driven fault detection and identification methods are important in large-scale chemical processes. However, some traditional methods often fail to show superior performance owing to the self-limitations and the characteristics of process data, such as nonlinearity, non-Gaussian distribution, and multi-operating mode. To cope with these issues, the k-NN (k-Nearest Neighbor) fault detection method and extensions have been developed in recent years. Nevertheless, these methods are primarily used for fault detection, and few papers can be found that examine fault identification. In this paper, in order to extract effective fault information, the relationship between various faults and abnormal variables is studied, and an accurate "fault⁻symptom" table is presented. Then, a novel fault identification method based on k-NN variable contribution and CNN data reconstruction theories is proposed. When there is an abnormality, a variable contribution plot method based on k-NN is used to calculate the contribution index of each variable, and the feasibility of this method is verified by contribution decomposition theory, which includes a feasibility analysis of a single abnormal variable and multiple abnormal variables. Furthermore, to identify all the faulty variables, a CNN (Center-based Nearest Neighbor) data reconstruction method is proposed; the variables that have the larger contribution indices can be reconstructed using the CNN reconstruction method in turn. The proposed search strategy can guarantee that all faulty variables are found in each sample. The reliability and validity of the proposed method are verified by a numerical example and the Continuous Stirred Tank Reactor system.

Predicting novel genes and pathways associated with osteosarcoma by using bioinformatics analysis.

This aim of this study was to explore novel biomarkers related to osteosarcoma. The mRNA expression profile GSE41293 dataset was downloaded from the Gene Expression Omnibus (GEO) database, which included seven osteosarcoma and six control samples. After preprocessing, the FASTQ format reads of 13 samples were mapped to the reference sequences to screen for unique mapping reads. Differentially expressed genes (DEGs) were selected, which were then used for pathway and protein-protein interaction (PPI) network analyses. Moreover, the microarray data GSE63631 were downloaded from GEO database to verify our results. The percentages of unique mapping reads for osteosarcomas and control samples were both >85%. A total of 6157 DEGs were identified between the two groups. DEGs that were upregulated were significantly enriched in 19 pathways, and those that were downregulated were enriched in 14 pathways. In the PPI network, DEGs such as SRC, ERBB2, and CAV3 in cluster 1 were enriched in the pathway responsible for focal adhesions. The DEGs in cluster 2, such as CDK4 and CDK6, were enriched in the cell cycle pathway. In GSE63631, DEGs were significantly enriched in focal adhesion pathway, which was in accordance with the result in GSE41293. Thus, the focal adhesion and cell cycle pathways may play important roles in osteosarcoma progression, and SRC, ERBB2, CAV3, CDK4, and CDK6 may be used as critical biomarkers of osteosarcoma.

Effects of Electrode Material on the Voltage of a Tree-Based Energy Generator.

The voltage between a standing tree and its surrounding soil is regarded as an innovative renewable energy source. This source is expected to provide a new power generation system for the low-power electrical equipment used in forestry. However, the voltage is weak, which has caused great difficulty in application. Consequently, the development of a method to increase the voltage is a key issue that must be addressed in this area of applied research. As the front-end component for energy harvesting, a metal electrode has a material effect on the level and stability of the voltage obtained. This study aimed to preliminarily ascertain the rules and mechanisms that underlie the effects of electrode material on voltage. Electrodes of different materials were used to measure the tree-source voltage, and the data were employed in a comparative analysis. The results indicate that the conductivity of the metal electrode significantly affects the contact resistance of the electrode-soil and electrode-trunk contact surfaces, thereby influencing the voltage level. The metal reactivity of the electrode has no significant effect on the voltage. However, passivation of the electrode materials markedly reduces the voltage. Suitable electrode materials are demonstrated and recommended.

[Correlation research on the MRI quantity of lumbar modic changes and low back pain].

OBJECTIVE: To analyzed the relationship between lumbar endplate Modic area changes rate and low back pain by measuring MRI T2 sagittal image of lumbar endplate Modic area changes rate. METHODS: From December 2011 to June 2012,70 patients with low back pain in operation were evaluated on pain by VAS and function by JOA,and examined by MRI including 39 males and 31 females with an average age of (51.00 +/- 11.89) years ranging from 29 to 72 years old. Among them, 54 cases had lumbar endplate Modic changes involving 15 cases in types Modic I ,21 cases in type Modic II, 11 cases in type Modic III ,mixed type Modic in 7 cases (eliminated for too few cases). Modic area changes and corresponding vertebral area were measured on MRI T2 median sagittal. The areas of two ways were compared to yield the rate of changes for Modic, for multisegmental Modic changes to calculate the total ratios. A correlation was observed among JOA, VAS and the rate of Modic changes. RESULTS: The correlation coefficient of change rate of Modic I with JOA score was r = -0.308, P = 0.048 < 0.05, there was a negative correlation;the correlation coefficient of change rate of Modic I with VAS scores was r = 0.428,P = 0.021 < 0.05, there was a positive correlation. The correlation coefficient of change rate of Modic II with JOA score was r = -0.375, P = 0.043 < 0.05, there was a negative correlation;the correlation coefficient of change rate of Modic II with VAS score was r = 0.352, P = 0.041 < 0.05, there was a positive correlation. The area change rate of Modic III had no significant correlation with low back pain degree (P > 0.05). CONCLUSION: Modic I and II area changes rate of of patients with low back pain is closely related to the degree of pain low back pain, Modic III area changes rate is not significant correlated to the degree of lower back pain.

[Polymorphism analysis of Plasmodium falciparum histidine-rich protein II and III].

OBJECTIVE: To analyze the polymorphism of Plasmodium falciparum histidine-rich protein (PfHRP) II and III. METHODS: Genomic DNA was isolated from blood samples of 20 patients infected with Plasmodium falciparum in Yunnan Province. Blood samples were tested by microscopy and RDTs. The Pfhrp2 and Pfhrp3 gene fragments were amplified by PCR and sequenced. The sequencing results were analyzed and compared using the bioinformatics software. RESULTS: 20 patients infected with Plasmodium falciparum tested by microscopy and RDTs. PCR showed that the Pfhrp2 gene was with 389~986 bp, and Pfhrp3 gene with 329-640 bp. All PfH-IRP II sequences started with type 1 repeat (AHHAHHVAD) and ended with the type 12 repeat (AHHAAAHHEAATH). The number of type 7 (AHHAAD), type 2 (AHHAHHAAD) and type 6 (AHHATD) within PfHRP II was more than the other types of repeats, as well as type 16 (AHHAAN) and type 17 (AHHDG) for PfHRP III. Type 11 repeat (AHN) was not found from the PfHRP II and PfHRP III sequences. CONCLUSION: There is an extensive diversity in Pfhrp2 and Pfhrp3 fragments in the individuals infected with P. falciparum in Yunnan. Some types of repeats are shared by PfHRP II and PfHRP III.

Testing the quality of images for permanent magnet desktop MRI systems using specially designed phantoms.

Our aim was to measure the performance of desktop magnetic resonance imaging (MRI) systems using specially designed phantoms, by testing imaging parameters and analysing the imaging quality. We designed multifunction phantoms with diameters of 18 and 60 mm for desktop MRI scanners in accordance with the American Association of Physicists in Medicine (AAPM) report no. 28. We scanned the phantoms with three permanent magnet 0.5 T desktop MRI systems, measured the MRI image parameters, and analysed imaging quality by comparing the data with the AAPM criteria and Chinese national standards. Image parameters included: resonance frequency, high contrast spatial resolution, low contrast object detectability, slice thickness, geometrical distortion, signal-to-noise ratio (SNR), and image uniformity. The image parameters of three desktop MRI machines could be measured using our specially designed phantoms, and most parameters were in line with MRI quality control criterion, including: resonance frequency, high contrast spatial resolution, low contrast object detectability, slice thickness, geometrical distortion, image uniformity and slice position accuracy. However, SNR was significantly lower than in some references. The imaging test and quality control are necessary for desktop MRI systems, and should be performed with the applicable phantom and corresponding standards.

Bone metastasis in a novel breast cancer mouse model containing human breast and human bone.

In practice, investigations for bone metastasis of breast cancer rely heavily on models in vivo. Lacking of such ideal model makes it difficult to study the whole process or accurate mechanism of each step of this metastatic disease. Development of xenograft mouse models has made great contributions in this area. Currently, the best animal model of breast cancer metastasizing to bone is NOD/SCID-hu models containing human bone, which makes it possible to let the breast cancer cells and the bone target of osteotropic metastasis be both of human origin. We have developed a novel mouse model containing both human bone and breast, and proved it functional and reliable. In this study, a set of human breast cancer cell line including MDA-MB-231, MDA-MB-231BO, MCF-7, ZR-75-1 and SUM1315 were characterized their osteotropism in this model. A specific cell line SUM1315 made species-specific bone metastasis, certifying the osteotropism-identification utility of the novel mouse model. Furthermore, gene expression and microRNA expression profiling analysis were done to the two SUM1315 derived sub lines isolated and purified from the orthotopic and metastatic xenograft. In addition, to demonstrate the disparity between the "spontaneous" and "forced" bone metastasis in mouse model, MDA-MB-231 cells were inoculated into both the human implants in this model simultaneously, and then primary cultured and profiling analyzed. Supported by overall results of profiling analyses, this study suggested the novel model was a useful tool for understanding, preventing and treating bone metastasis of breast cancer, meanwhile it had provided significant information for further investigations.

STAT3 mediates resistance of CD44(+)CD24(-/low) breast cancer stem cells to tamoxifen in vitro.

We sought to determine whether STAT3 mediated tamoxifen resistance of breast cancer stem cells in vitro. The capacities for mammosphere formation and STAT3 expression of CD44(+)CD24(-/low) MCF-7 and MCF-7 were observed. The CD44(+)CD24(-/low) subpopulation ratio and its sensitivity to adriamycin were analyzed in MCF-7 and TAM resistant (TAM-R) cells. Cell cycle, apoptosis, STAT3 and phospho-STAT3 changes were observed after treatment with tamoxifen. Small interference RNA-mediated knockdown of STAT3 in TAM-R cells was also performed. CD44(+)CD24(-/low) MCF-7 showed higher capacities for mammosphere formation and STAT3 expression than total MCF-7. The CD44(+)CD24(-/low) subpopulation was also upregulated in TAM-R cells with less sensitivity to adriamycin than MCF-7. Cell cycle changes, anti-apoptotic effects and STAT3 changes were also found. Meanwhile, the knock-down of STAT3 in TAM-R resulted in an increase in sensitivity to tamoxifen. It is concluded that STAT3 plays an essential role in breast cancer stem cells, which correlated with tamoxifen resistance.