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This study aimed to investigate the optimal tension for the reconstruction of the distal radioulnar ligaments (DRULs) in the treatment of the distal radioulnar joint (DRUJ) instability.A total of eight human cadaver upper extremities were used. First, the Tekscan sensor film system was used to measure the contact characteristics of the intact DRUJ. Following this, the DRULs were resected, and the measurement was repeated. The DRULs were then reconstructed according to Adams' procedure, and the contact forces under different initial tension were compared with that of the intact group to obtain the optimal tension. At that point, the contact force of the DRUJ was close to normal. The reliability of the obtained tension was verified by translational testing, which reflected the stability of the DRUJ.In the neutral position, the contact force, area, and pressure inside DRUJ were 0.51 ± 0.10 N, 64.08 ± 11.58 mmBy comparing the contact forces under different reconstruction tensions with the normal value, we obtained the optimal tension, which can provide the theoretical basis for the clinical treatment of chronic DRUJ instability.

Breast cancer remains a dangerous disease, and delving the molecular mechanism of breast cancer is still necessary. To illustrate the role of miR-511-5p, TCGA database was used to excavate the abundance of miR-511-5p, and the miR-511-5p level was measured in the pathological tissues and tumor cell lines. Moreover, the targets of miR-511-5p were identified with miRDIP and GEPIA and then were used for functional enrichment analysis. Besides, the targets of miR-511-5p were analyzed with the protein-protein interaction (PPI) network for the hub nodes, and then the expression levels of the hub nodes were visualized with the GEPIA database. The results showed that miR-511-5p was significantly downregulated in multiple types of tumor samples in the online database, and the downregulated miR-511-5p was also found in pathological tissues and tumor cell lines. Moreover, 48 genes were identified as the potential targets of miR-511-5p by miRDIP and GEPIA databases and enriched in cell cycle, PI3K/AKT, and P53 pathways. Besides, seven genes including BRCA1, FN1, CCNE1, CCND1, CHEK1, BUB3, and CDC25A were identified as the hub nodes by the PPI network, and CCNE1 and CHEK1 were confirmed to be related with the prognostic survival of the patients with breast cancer. In conclusion, the proofs in this study suggest that reduced miR-511-5p was a biomarker event for breast cancer, and CCNE1 and CHEK1 served as potential targets of miR-511-5p to involve the progression of breast cancer.

Transforming growth factor β (TGFβ) is prometastatic in advanced cancers and its biological activities are mainly mediated by the Smad family of proteins. Smad4 is the central signal transducer and transcription factor in the TGFβ pathway, yet the underlying mechanisms that govern transcriptional activities of Smad4 are not fully understood. Here, we show that AMBRA1, a member of the DDB1 and CUL4-associated factor (DCAF) family of proteins, serves as the substrate receptor for Smad4 in the CUL4-RING (CRL4) ubiquitin ligase complex. The CRL4-AMBRA1 ubiquitin ligase mediates nonproteolytic polyubiquitylation of Smad4 to enhance its transcriptional functions. Consequently, AMBRA1 potentiated TGFβ signaling and critically promoted TGFβ-induced epithelial-to-mesenchymal transition, migration, and invasion of breast cancer cells. Mouse models of breast cancer demonstrated that AMBRA1 promotes metastasis. Collectively, these results show that CRL4-AMBRA1 facilitates TGFβ-driven metastasis by increasing Smad4 polyubiquitylation, suggesting AMBRA1 may serve as a new therapeutic target in metastatic breast cancer. Significance: This study identifies AMBRA1 as a novel regulator of TGFβ signaling and breast cancer metastasis, supporting further exploration of AMBRA1 as a target for cancer therapy.

BACKGROUND The tp0548 gene, hypothesized to encode for an outer-membrane protein, was originally used in the enhanced Centers for Disease Control and Prevention typing for molecular typing of Treponema pallidum. It plays an important role in the molecular epidemiology of Treponema because it is not only an important locus of multiple typing approaches but also suitable for strain typing of multiple Treponema subspecies. METHODS A 27-year-old Chinese man attended the Institute of Dermatology, Chinese Academy of Medical Sciences Sexually Transmitted Disease Clinic in Nanjing, China, because of a genital ulcer and inguinal lymphadenopathy for 1 week. Workup consisted of microbiological and hematological investigations, and sequences analysis. The aims of this study were to describe a novel tp0548 sequence type "Qn" of this syphilis strain and to review all previously reported novel tp0548 genotypes. RESULTS We identified a novel tp0548 gene type in a genital ulcer in a patient with primary syphilis in Nanjing, China. Using sequence alignment, we further found that this novel sequence was closely similar to "Q." Following the nomenclature used in the enhanced Centers for Disease Control and Prevention typing methodology, the letters "Qn" was assigned to the new sequence type. CONCLUSION The novel tp0548 sequence type of T. pallidum not only expands the database up to 27 different sequence types but also indicates the substantial genetic diversity of the tp0548 gene sequence.

This study aimed to explore the expression of lncRNA-metastasis associated lung adenocarcinoma transcript 1 (lncRNA-MALAT1) in breast cancer (BC) patients and its influences on the prognosis of the patients. A total of 120 BC patients admitted to our hospital were enrolled as a BC group, of which 58 patients at I/II stage were treated with breast-conserving surgery as an operation group, and the other 62 patients at III/IV stage were treated with neoadjuvant chemotherapy combined with breast-conserving surgery as a combination group. Meantime, 100 healthy people in physical examination during the same period were enrolled as a normal group. The expression of serum lncRNA-MALAT1 in the subjects was determined, and the expression in BC patients and its influences on the patients were analyzed. LncRNA-MALAT1 was over-expressed in patients from the BC group, and the area-under-the-curve (AUC) of it for diagnosing BC was 0.911. After treatment, the expression of lncRNA-MALAT1 in the operation group and the combination group significantly decreased, and the expression of it in patients with good prognosis was greatly lower than that in patients with poor prognosis. The AUC of lncRNA-MALAT1 for predicting poor prognosis was 0.838, and TNM staging, pathological differentiation, tumor diameter, and lncRNA-MALAT1 were independent prognostic factors for poor prognosis of the patients. Furthermore, low expression of lncRNA-MALAT1 was associated with a relatively high 5-year overall survival (OS) of BC patients. The expression of lncRNA-MALAT1 was up-regulated in BC patients, while it was down-regulated in BC patients treated with breast-conserving surgery combined with neo-adjuvant chemotherapy, so lncRNA-MALAT1 can be used as a potential indicator for early diagnosis and prognosis prediction of BC patients.

Herein, a label-free and homogeneous electrochemical strategy for monitoring of matrix metalloproteinase 2 (MMP-2) activity was proposed based on electrodes modified with orderly distributed mesoporous silica films (MSFs). In the absence of target MMP-2, an artificially substrate peptide with positive charge was absorbed on the surface of MSFs by electrostatic interaction, which could prevent electrochemical molecules [Ru(NH3)6]Cl3 from approaching the electrode surface. When the substrate peptide was hydrolyzed by target MMP-2, [Ru(NH3)6]Cl3 could arrive to the electrode surface and lead to the increase of electrochemical signal. This assay showed considerable sensitivity to target MMP-2, which could measure it down to 0.98 ng. mL−1. Meanwhile, a satisfied response to the inhibitor of MMP-2 was also achieved (IC-50 value = 1.68 μM). Significantly, it displayed satisfactory performances in the complicated biological samples including cell lysates and human serum. Taking advantages of the anti-fouling ability in biological complex samples of MSFs and the high efficiency of homogeneous sensing, this assay realized the electrochemical detection of MMP-2 with accuracy and sensitivity, which exhibited significant potential in clinical biomedicine and biological analysis of cancer-related protease.

Smads are critical intracellular signal transducers for transforming growth factor-β (TGF-β) in mammalian cells. In this study, we have identified WD repeat-containing protein 74 (WDR74) as a novel transcriptional coactivator for Smads in the canonical TGF-β signaling pathway. Through direct interactions with Smad proteins, WDR74 enhances TGF-β-mediated phosphorylation and nuclear accumulation of Smad2 and Smad3. Consequently, WDR74 enables stronger transcriptional responses and more robust TGF-β-induced physiological responses. Our findings have elucidated a critical role of WDR74 in regulating TGF-β signaling.

Presented herein is a simple, robust, and label-free homogeneous electrochemical sensing platform constructed for the detection of protein kinase activity and inhibition by integration of carboxypeptidase Y (CPY)-assisted peptide cleavage reaction and vertically ordered mesoporous silica films (MSFs). In this sensing platform, the substrate peptide composed of kinase-specific recognized sequence and multiple positively charged arginine (R) residues was ingeniously designed. In the presence of protein kinase, the substrate peptide was phosphorylated and then immediately resisted CPY cleavage. The phosphorylated peptide could be effectively adsorbed on the negatively charged surface of MSFs modified indium-tin oxide (ITO) electrode (MSFs/ITO) by noncovalent electrostatic attraction. The adsorbed peptide was subsequently used as a hamper to prevent the diffusion of electroactive probe (FcMeOH) to the electrode surface through the vertically aligned nanopores, resulting in a detectable reduction of electrochemical signal. As demonstrated for the feasibility and universality of the sensing platform, both protein kinase A (PKA) and casein kinase II (CK2) were selected as the models, and the detection limits were determined to be 0.083 and 0.095 UmL

In this work, based on mesoporous silica containers (MSNs) with the programmed enzyme-free DNA assembly amplification of catalytic hairpin assembly (CHA) and hybridization chain reaction (HCR), an ultrasensitive electrochemical sensing platform with low background is developed for the detection of microRNA (miRNA). Herein, the electrochemical reporter methylene blue (MB) was sealed in the pores of MSNs by the double-stranded DNA (dsDNA) gate of hairpin DNA H1 and anchor DNA. In the absence of target, neither the CHA nor the HCR process happened, which enabled a low background. After target was added, DNA H1 was displaced from the MSNs surface and participated in the CHA process with the assistance of hairpin DNA H2, which accelerated the release of MB from the MSNs pore. Meanwhile, the CHA products H1-H2 were hybridized with the capture probes (SH-CP) on the electrode surface, which further initiated the HCR process. The released MB from the MSNs will effectively intercalate into long dsDNA polymers of HCR products, resulting in a significant electrochemical response. Taking miRNA-21 as the model target, the proposed sensing platform achieves a satisfactory detection limit down to 0.037 fM, which is lower than that of electrochemical assay with amplification methods. In addition, the strategy shows good selectivity against other miRNAs and is capable in practical analytes. Benefitting from the features of being label-free and enzyme-free and having low background, high sensitivity, and selectivity, this strategy shows great potential in bioanalysis and clinical diagnostics.

Smad proteins are central mediators in the canonical transforming growth factor-β (TGF-β) signaling pathway in mammalian cells. We report here that bromodomain-containing protein 7 (BRD7) functions as a novel transcription coactivator for Smads in TGF-β signaling. BRD7 forms a TGF-β inducible complex with Smad3/4 through its N-terminal Smad-binding domain. BRD7 simultaneously binds to acetylated histones to promote Smad-chromatin association, and associates with histone acetyltransferase p300 to enhance Smad transcriptional activity. Ectopic expression of BRD7, but not its mutants defective in Smad binding, enhances TGF-β transcriptional, tumor suppressing and epithelial-mesenchymal transition (EMT) responses. Conversely, depletion of BRD7 inhibits TGF-β responses. Thus, our study provides compelling evidence for a new function of BRD7 in fine-tuning TGF-β physiological responses.

Exosomal microRNAs (miRNAs) transferred between cells have been implicated in modulating the host immune response in microbial infections. In this study, we isolated exosomes from Treponema pallidum (T. pallidum)-stimulated macrophages and detected differential exosomal miRNA expression using both microarrays, and RT-qPCR. A total of 65 differentially expressed miRNAs (35 upregulated and 30 downregulated) were identified. Of all identified miRNAs, miR-146a-5p was one of the most significantly changed miRNAs with high expression in exosomes from T. pallidum-stimulated macrophages. Furthermore, we isolated plasma exosomes from early syphilis patients and healthy controls, and confirmed miR-146a-5p upregulation in the former group. We also show that exosomal miR-146a-5p is efficiently transported into endothelial cells, reducing monocyte transendothelial migration and endothelial permeability by targeting junctional adhesion molecule C (JAM-C). Luciferase reporter assays confirmed binding of exosomal miR-146a-5p to the 3'untranslated region (3'UTR) of JAM-C. We then demonstrated that also exosomes derived from macrophages stimulated by T. pallidum expressed high levels of miR-146a-5p which could be delivered to endothelial cells, and decreased monocyte transendothelial migration by targeting JAM-C. Overall, this work provides novel insights into the mechanism by which T. pallidum hampers inflammatory reactions of the host via a blockade of leukocytes transendothelial migration and endothelial permeability.

PROBLEM IDENTIFICATION Fear of cancer recurrence (FCR) is prevalent among people with cancer; therefore, it is important to determine whether interventions offered to this population are effective in alleviating FCR. This review summarizes randomized, controlled trials in the literature about the effect of psychosocial interventions on FCR in people with cancer. LITERATURE SEARCH A systematic review of the literature was conducted using PubMed, Web of Science, EMBASE, and Cochrane Library. DATA EVALUATION Extracted data included cancer type, number of participants, intervention type and duration, follow-ups, and primary and secondary outcomes. SYNTHESIS 10 studies met inclusion criteria. Types of interventions included mindfulness-based therapy (n = 3), cognitive behavioral therapy (n = 4), psychoeducational interventions (n = 1), gratitude interventions (n = 1), and communication interventions (n = 1). IMPLICATIONS FOR PRACTICE Psychosocial interventions are beneficial for alleviating FCR and positively affect depression, anxiety, and quality of life in people with cancer.

Herein a low background cascade signal amplification electrochemical sensing platform has been proposed for the ultrasensitive detection of mRNA (mRNA) by coupling the target-activated hybridization chain reaction and electroactive cargo release from mesoporous silica nanocontainers (MSNs). In this sensing platform, the 5'-phosphate-terminated DNA (5'-PO4 cDNA) complement to target mRNA is hybridized with the trigger DNA and anchor DNA on the surface of the MSNs, aiming at forming a double-stranded DNA gate molecule and sealing the methylene blue (MB) in the inner pores of the MSNs. In the presence of target mRNA, the 5'-PO4 cDNA is displaced from the MSNs and competitively hybridizes with mRNA, which led to the liberation of the trigger DNA and the opening of the MSNs pore. The liberated trigger DNA can be then immobilized onto the electrode surface through hybridization with the capture DNA, triggering HCR on the electrode surface. At the same time, the MB released from the MSNs will selectively intercalate into the HCR long dsDNA polymers, giving rise to significant electrochemical response. In addition, due to the λ-exonuclease (λ-Exo) cleavage reaction-assisted target recycling, more amounts of trigger DNA will be liberated and trigger HCR, and numerous MB are uncapped and intercalate into the HCR products. As proof of concept, thymidine kinase 1 (TK1) mRNA was used as a model target. Featured with amplification efficiency, label-free capability, and low background signal, the strategy could quantitatively detect TK1 mRNA down to 2.0 aM with a linear calibration range from 0.1 fM to 1 pM. We have also demonstrated the practical application of our proposed sensing platform for detecting TK1 mRNA in real samples, opening up new avenues for highly sensitive quantification of biomarkers in bioanalysis and clinical diagnosis.

TGF-β controls a variety of cellular functions during development. Abnormal TGF-β responses are commonly found in human diseases such as cancer, suggesting that TGF-β signaling must be tightly regulated. Here, we report that protein tyrosine phosphatase non-receptor 3 (PTPN3) profoundly potentiates TGF-β signaling independent of its phosphatase activity. PTPN3 stabilizes TGF-β type I receptor (TβRI) through attenuating the interaction between Smurf2 and TβRI. Consequently, PTPN3 facilitates TGF-β-induced R-Smad phosphorylation, transcriptional responses, and subsequent physiological responses. Importantly, the leucine-to-arginine substitution at amino acid residue 232 (L232R) of PTPN3, a frequent mutation found in intrahepatic cholangiocarcinoma (ICC), disables its role in enhancing TGF-β signaling and abolishes its tumor-suppressive function. Our findings have revealed a vital role of PTPN3 in regulating TGF-β signaling during normal physiology and pathogenesis.

The epithelial-to-mesenchymal transition (EMT) is a cellular reprogramming process converting epithelial cells into mesenchymal cell morphology. Snail is a critical regulator of EMT by both suppressing epithelial gene expression and promoting mesenchymal gene expression. Expression and activity of Snail are tightly controlled at transcriptional and post-translational levels. It has previously been reported that Snail undergoes phosphorylation and ubiquitin-dependent proteasome degradation. Here, we report nuclear phosphatase SCP4/CTDSPL2 acts as a novel Snail phosphatase. SCP4 physically interacts with and directly dephosphorylates Snail. SCP4-mediated dephosphorylation of Snail suppresses the ubiquitin-dependent proteasome degradation of Snail and consequently enhances TGFβ-induced EMT. The knockdown of SCP4 in MCF10A mammary epithelial cells leads to attenuated cell migration. Collectively, our finding demonstrates that SCP4 plays a critical role in EMT through Snail dephosphorylation and stabilization.

A highly sensitive electrochemiluminescence (ECL) strategy was developed for the protein kinase A (PKA) activity and inhibition assay based on double-quenching of graphene quantum dots (GQDs) ECL by G-quadruplex-hemin DNAzyme and gold nanoparticles (AuNPs). In this strategy, the GQDs were modified onto the indium-tin oxide (ITO) electrode and further assembled with substrate peptide of target protein kinase through covalent coupling, which can exhibit high and stable ECL signal. The AuNPs, functionalized with the phosphorylated DNA and G-quadruplex-hemin DNAzyme via Au-S chemistry, were selected as quenching probes. In the presence of PKA, the peptide on the electrode was phosphorylated and the AuNPs functionalized with the phosphorylated DNA and G-quadruplex-hemin DNAzyme were subsequently integrated onto the phosphorylated peptide by Zr(4+). Owing to the reduction of coreactant H2O2 resulting from G-quadruplex-hemin DNAzyme catalytic reaction and the ECL energy transfer (ECL-RET) between AuNPs and GQDs, the ECL intensity of GQDs was significantly decreased. By taking advantage of the double-quenching effect, this assay can detect PKA with a linear range of 0.05 to 5 U mL(-1) and a detection limit of 0.04 U mL(-1). In addition, the PKA inhibition assay and interferences experiments of CK2 and T4 PNK have been studied respectively. This assay was also successfully applied to PKA assay in serum samples and cell lysates, indicating that the developed method have the potential applications in protein kinase-related biochemical fundamental research and clinical diagnosis.

A sensitive and simple electrochemical strategy has been developed for assay of protein kinase A (PKA) activity and inhibition using gold nanoparticles/multi-walled carbon nanotubes (AuNPs/MWNTs) nanohybrids. Key features of this assay included intrinsic peroxidase-like activity of positively-charged gold nanoparticles (+AuNPs) and signal transduction and amplification of multi-walled carbon nanotubes (MWNTs). In this assay, an N-terminally cysteine-containing peptide was self-assembled onto the gold electrode via Au–S bonding and used as substrate for PKA, and adenosine-5′-(γ-thio)-triphosphate was used as co-substrate. Upon thiophosphorylation in the presence of PKA, the AuNPs/MWNTs nanohybrids would be fixed onto the peptides via Au–S bond. The conjugated AuNPs/MWNTs nanohybrids could catalyze the 3, 3′, 5, 5′-Tetramethylbenzidine (TMB) oxidation by H2O2 to form TMB oxidation product, which was reduced at the electrode surface to generate an electrochemical current. It was eT on state. The current signal intensity is proportional to the activity of PKA. Here, the presence of MWNTs not only increased the surface area for accumulation of +AuNPs but also could promote electron-transfer reaction. It was found that the electrochemical strategy can be employed to assay PKA activity with a low detection limit of 0.09 U/mL. The linear range of the assay for PKA enzymatic unit/ml was 0.1–1 U/mL. Furthermore, the interferences experiments of T4 polynucleotide kinase (T4 PNK) and Casein kinase II (CK2), and inhibition of PKA, have also been studied by using this strategy. The developed method would provide a diversified platform for kinase activity and inhibition monitoring.

DNA repair processes are responsible for maintaining genome stability. Ligase and polynucleotide kinase (PNK) have important roles in ligase-mediated DNA repair. The development of analytical methods to monitor these enzymes involved in DNA repair pathways is of great interest in biochemistry and biotechnology. In this work, we reported a new strategy for label-free monitoring PNK and ligase activity by using dumbbell-shaped DNA templated copper nanoparticles (CuNPs). In the presence of PNK and ligase, the dumbbell-shaped DNA probe (DP) was locked and could resist the digestion of exonucleases and then served as an efficient template for synthesizing fluorescent CuNPs. However, in the absence of ligase or PNK, the nicked DP could be digested by exonucleases and failed to template fluorescent CuNPs. Therefore, the fluorescence changes of CuNPs could be used to evaluate these enzymes activity. Under the optimal conditions, highly sensitive detection of ligase activity of about 1U/mL and PNK activity down to 0.05U/mL is achieved. To challenge the practical application capability of this strategy, the detection of analyte in dilute cells extracts was also investigated and showed similar linear relationships. In addition to ligase and PNK, this sensing strategy was also extended to the detection of phosphatase, which illustrates the versatility of this strategy.

Measuring pH in living cells is of great importance for better understanding cellular functions as well as providing pivotal assistance for early diagnosis of diseases. In this work, we report the first use of a novel kind of label-free carbon dots for intracellular ratiometric fluorescence pH sensing. By simple one-pot hydrothermal treatment of citric acid and basic fuchsin, the carbon dots showing dual emission bands at 475 and 545 nm under single-wavelength excitation were synthesized. It is demonstrated that the fluorescence intensities of the as-synthesized carbon dots at the two emissions are pH-sensitive simultaneously. The intensity ratio (I475 nm/I545 nm) is linear against pH values from 5.2 to 8.8 in buffer solution, affording the capability as ratiometric probes for intracellular pH sensing. It also displays that the carbon dots show excellent reversibility and photostability in pH measurements. With this nanoprobe, quantitative fluorescence imaging using the ratio of two emissions (I475 nm/I545 nm) for the detection of intracellular pH were successfully applied in HeLa cells. In contrast to most of the reported nanomaterials-based ratiometric pH sensors which rely on the attachment of additional dyes, these carbon-dots-based ratiometric probes are low in toxicity, easy to synthesize, and free from labels.

OBJECTIVE: To prospectively investigate the association between serum uric acid (SUA) level and incidence of impaired fasting glucose (IFG) in adult Chinese. METHODS: We evaluated 13,328 women and 41,350 men without diabetes and IFG. The participants were classified into quintile according to baseline level of SUA. Data were analyzed to examine the association between SUA levels and the incidence of IFG. We used Cox regression models to estimate the relative risk of IFG after adjusting for known risk factors. RESULTS: For men, the second quintile of SUA has the lowest cumulative incidence of IFG (29.9%); the fifth quintile of SUA has the highest cumulative incidence of IFG (35.6%). After corrected with Cox regression, the first quartile and the fourth quartile have higher cumulative incidence of IFG than the second quintile, with the HR of 1.11(1.05-1.17) and 1.07(1.01-1.13), respectively. For women, the first quartile of SUA has the lowest cumulative incidence of IFG (20.7%), while the fifth quintile of SUA has the highest cumulative incidence of IFG (30.0%). However, there is no significant difference in IFG between different quintile after adjusted with Cox regression. CONCLUSIONS: The results of this prospective study suggest that there is a higher risk of developing IFG in association with low or high SUA concentrations for men. These relationships were independent of other known risk factors. There is no significant correlation in the risk of developing IFG in association with SUA concentrations for women. Analyses excluding participants with hypertension or with hyperlipidemia and analyses with participants stratified by age reached similar conclusion.