Protein-activated and FRET enhanced excited-state intermolecular proton transfer fluorescent probes for high-resolution imaging of cilia and tunneling nanotubes in live cells.

Excited-state intermolecular proton transfer (inter-ESPT) fluorescent probes responsive to specific bioactive molecules should be greatly promising for protein sensing, DNA mutation simulating and cellular process regulating. However, the inter-ESPT molecules are recessive ESPT fluorophores, which need the assistance of other molecules with both hydrogen-bond accepting and donating abilities to turn on the tautomeric fluorescence. Valid design strategies to create powerful inter-ESPT fluorescent probes are poorly developed, particularly for proteins as targets. We recently reported a unique supramolecular strategy to trigger the inter-ESPT process based on the probe-protein recognition by H-bonding and to image protein-based subcellular structures in live cells. Herein, we found that our inter-ESPT probes (inter-ESPT-01) bearing a 2-amino-3-cyanopyridine scaffold can anchor proteins and light up the "invisible" ESPT state, so as to image the proteins or the protein-based subcellular organelles. More importantly, the inter-ESPT emission of inter-ESPT-01 can be significantly enhanced by the FRET process between amino and imino tautomers, endowing the inter-ESPT-01 probes with super-bright tautomeric fluorescence. The expressed proteins Ecallantide and MarTX were selected as the models to light up the inter-ESPT fluorescence of the probes and revealed that the inter-ESPT process can be triggered by the specific probe-protein recognition events. In the use of the super-bright inter-ESPT fluorescence, not only the proteins, but also the protein-based cilia and tunneling nanotubes (TNTs) can be specifically visualized in living cancer cells. Furthermore, such recognition-driven strategy allows us to construct a unique inter-ESPT probe to track and image specific endogenous proteins in live cells, highlighting the potential of inter-ESPT fluorogens as novel intelligent biomaterials.

The diagnostic and prognostic value of plasma circulating circNUP98 for nasopharyngeal carcinoma.

BACKGROUND: Our preliminary sequencing analysis revealed increased expression levels of circNUP98 in nasopharyngeal carcinoma (NPC). This study was therefore carried out to explore the role of circNUP98 in NPC. METHODS: The present study enrolled 56 patients with NPC, 44 patients with cervical lymphadenitis (CL), 50 patients with nose bleeding (NB), 50 patients with chronic sinusitis (CS), 50 patients with lymph node tuberculosis (LNT), and 50 healthy controls (Control). Plasma samples were obtained from all patients and the controls. In addition, NPC and paired non-tumor tissue samples were collected from the 56 NPC patients. Expression of circNUP98 in both tissue and plasma samples was determined by RT-qPCR. The 56 NPC patients were followed up for 5 years to analyze the associations between plasma expression of circNUP98 and the survival of patients. The diagnostic value of circNUP98 for NPC was analyzed through ROC curve analysis. RESULTS: The plasma expression levels of circNUP98 were only increased in NPC, but not in CL, NB, CS and LNT groups compared to that in the Control group. In addition, increased expression levels of circNUP98 was observed in NPC tissues compared to that in non-tumor tissues. Plasma circNUP98 was closely correlated with circNUP98 in NPC tissues, but not circNUP98 in non-tumor tissues. With plasma circNUP98 as a biomarker, NPC patients were separated from CL, NB, CS, LNT and the Control groups. Plasma expression of circNUP98 was found to be positively correlated with the poor survival of patients. Moreover, plasma circNUP98 was only closely associated with tumor metastasis, but not tumor size. CONCLUSION: The expression of circNUP98 in plasma may be used to diagnose NPC and predict patients' survival.

WNT1 expression influences the development of dysplasia of the hip via regulating RBPMS2/NOG-BMP2/4-GDF5- WISP2 pathway.

OBJECTIVE: To explore the role of WNT family member 1 (WNT1) in the development of dysplasia of the hip (DDH) and the molecular mechanism involved in this process. Methods: Si-WNT1, pcDNA3.1-WNT1 or corresponding negative controls were transfected into human osteoblast hFOB1.19 and human chondrocyte C28/I2, respectively. The proliferation of cells was measured by EdU assay. The relative expressions of human noggin gene (NOG), growth differentiating factor 5 (GDF5), WNT1, and WNT1-inducible-signaling pathway protein 2 (WISP2) were determined by immunofluorescence analysis. The protein expressions of RNA-binding protein of multiple splice forms 2 (RBPMS2), NOG, bone morphogenetic protein 2 (BMP2), BMP4, WNT1 and WISP2 were determined by western blot. Animal experiment was also performed and the morphological development of hip joint was observed. Results: Overexpression of WNT1 promoted osteoblast proliferation and inhibited chondrocyte proliferation, while knockdown of WNT1 inhibited osteoblast proliferation. In chondrocytes, knockdown of WNT1 upregulated NOG expression, while overexpression of WNT1 downregulated its expression. In osteoblasts and chondrocytes, overexpression of WNT1 increased BMP2, BMP4, WNT1, and WISP2 expression. RBPMS2 and NOG were slightly expressed in each group. Conclusion: Overexpression of WNT1 promoted osteoblast proliferation, inhibited chondrocyte proliferation, and increased the expressions of BMP2, BMP4, WNT1, and WISP2. Therefore, WNT1 may be a new therapeutic target for DDH.

Fluid Homeostasis May Predict the Prognosis of Non-infectious Fever After Total Knee Arthroplasty Within 7-Day: A Retrospective Cohort Study.

Background: In the perioperative management of Total Knee Arthroplasty (TKA), postoperative fever has always been a concern. Current research focuses on infectious fever, and there is no relevant research on the occurrence of non-infectious fever (NIF) and its risk factors. Hence, the aim of this study was to clarify the risk factors for NIF after TKA, and construct an easy-to-use nomogram. Methods: A retrospective cohort study was conducted. Consecutive patients undergoing primary unilateral TKA were divided into the non-infectious fever group and the control group. Clinicopathological characters were collected from electronic medical records. Univariate Logistic regression was used to analyze the related independent risk factors. The optimal threshold for each selected factor and combined index was determined when the Youden index achieved the highest value. And the predictive nomogram was developed by these independent factors. Results: Ultimately, 146 patients were included in this study. Of them, 57 (39.04%) patients experienced NIF. Results of the univariable logistic regression analysis indicated that intraoperative blood loss (OR, 1.002; 95% CI, 1.000-1.0004), postoperative drainage fluid volume (OR, 1.003; 95% CI, 1.001-1.006) and frequency of blood transfusion (n = 1; OR, 0.227; 95% CI, 0.068-0.757) were independent risk factors of NIF occurrence. The predictive nomogram that incorporated the above independent risk factors was developed, and it yielded an areas under the curves (AUC) of 0.731 (95% CI: 0.651-0.801; P < 0.0001) with 54.39% sensitivity and 82.02% specificity. Conclusions: Non-infectious fever after TKA prolongs the time of antibiotic use and hospital stay. Our results demonstrated that the nomogram may facilitate to predict the individualized risk of NIF occurrence within 7-day by intraoperative blood loss, postoperative drainage fluid volume and frequency of blood transfusion.

Monteggia equivalent lesion in children: a narrative review.

Background: Monteggia equivalent lesion represents a group of injury or combined injury patterns that resemble the Monteggia lesion in its presentations and mechanisms. Unlike Monteggia lesions, the equivalent ones, which share vague definitions and mostly occur as sporadic single case reports in the literature, have not been thoroughly reviewed since Bado first proposed the term, especially in the pediatric population. The objective of this review was to elucidate the definition by elaborating on its clinical styles and thus analyzing the mechanism, diagnosis, and management through related literature. Data sources: Based on the terms of 'Monteggia equivalent', 'radial neck fracture' and 'pediatric', all of the related literature was searched on the PubMed and Google Scholar search engine. Results: The advance of the definitions for pediatric Monteggia equivalent lesion (PMEL) was reviewed. The functional roles of the ulnar and the related mechanism theories in this injury were analyzed. The status of the radiocapitellar joint in this injury was emphasized. According to the previous statements, a new classification model was proposed and proper diagnosis and treatment approaches were suggested. Conclusions: PMEL should be defined as an ulnar fracture at any level combined with a proximal radial fracture. According to the status of the radiocapitellar joint, it could be divided into three groups. The occult ulnar bowing and delayed radial head dislocation should be a serious concern of orthopedists. Surgical need is usually warranted. Maintaining the ulnar length and securing the radiocapitellar joint are highly recommended.

Nucleosome Assembly Protein 1-Like 3 Enhances Cisplatin Resistance of Ovarian Cancer Cell by Activating Transforming Growth Factor-Beta Pathway.

Chemotherapy resistance is one of the main reasons for tumor-related death. In particular, ovarian cancer patients often acquire drug resistance after chemotherapy. In this study, we found that the histone chaperone, nucleosome assembly protein 1-like 3 (NAP1L3), was significantly upregulated in tissues with cisplatin resistance compared with cisplatin-sensitive tissues. Patients with high NAP1L3 levels had poor prognosis, suggesting that NAP1L3 might regulate ovarian cancer resistance. Colony formation and terminal deoxynulceotidyl transferase nick-end-labeling (TUNEL) assays showed cells with high NAP1L3 had high cisplatin resistance, whereas cells with low NAP1L3 had poor cisplatin resistance. NAP1L3 overexpression significantly increased cisplatin resistance, whereas NAP1L3 knockdown significantly reduced cisplatin resistance, suggesting that NAP1L3 promoted cisplatin resistance. Mechanistically, gene set enrichment analysis and luciferase reporter assays showed that NAP1L3 regulated the transforming growth factor-beta (TGF-ß) pathway. NAP1L3 overexpression increased the phosphorylation and nuclear translocation of SMAD family member 2 (SMAD2) and SMAD3, confirming that NAP1L3 activated the TGF-ß pathway. Therefore, NAP1L3 might represent a novel target to overcome ovarian cancer chemoresistance.

N-acetyl-cysteine attenuates neuropathic pain by suppressing matrix metalloproteinases.

The treatment of neuropathic pain remains a clinical challenge because of its unclear mechanisms and broad clinical morbidity. Matrix metalloproteinase (MMP)-9 and MMP-2 have previously been described as key components in neuropathic pain because of their facilitation of inflammatory cytokine maturation and induction of neural inflammation. Therefore, the inhibition of MMPs may represent a novel therapeutic approach to the treatment of neuropathic pain. In this study, we report that N-acetyl-cysteine (NAC), which is a broadly used respiratory drug, significantly attenuates neuropathic pain through a unique mechanism of MMP inhibition. Both the in vitro (0.1 mM) and in vivo application of NAC significantly suppressed the activity of MMP-9/2. Orally administered NAC (50, 100, and 200 mg/kg) not only postponed the occurrence but also inhibited the maintenance of chronic constrictive injury (CCI)-induced neuropathic pain in rats. The administration of NAC blocked the maturation of interleukin-1ß, which is a critical substrate of MMPs, and markedly suppressed the neuronal activation induced by CCI, including inhibiting the phosphorylation of protein kinase Cγ, NMDAR1, and mitogen-activated protein kinases. Finally, NAC significantly inhibited CCI-induced microglia activation but elicited no notable effects on astrocytes. These results demonstrate an effective and safe approach that has been used clinically to alleviate neuropathic pain through the powerful inhibition of the activation of MMPs.