Astrocytes are closely linked to depression, and the prefrontal cortex (PFC) is an important brain region involved in major depressive disorder (MDD). However, the underlying mechanism by which astrocytes within PFC contribute to MDD remains unclear. Using single-nucleus RNA sequencing analyses, we show a significant reduction in astrocytes and attenuated pleiotrophin-protein tyrosine phosphatase receptor type Z1 (PTN-PTPRZ1) signaling in astrocyte-to-excitatory neuron communication in the PFC of male MDD patients. We find reduced astrocytes and PTN in the dorsomedial PFC of male mice with depression induced by chronic restraint and social defeat stress. Knockdown of astrocytic PTN induces depression-related responses, which is reversed by exogenous PTN supplementation or overexpression of astrocytic PTN. The antidepressant effects exerted by astrocytic PTN require interaction with PTPRZ1 in excitatory neurons, and PTN-PTPRZ1 activates the AKT signaling pathway to regulate depression-related responses. Our findings indicate the PTN-PTPRZ1-AKT pathway may be a potential therapeutic target for MDD.

Lung transplantation results are compromised by ischemia–reperfusion injury and alloimmune responses. Ex vivo lung perfusion (EVLP) is used to assess marginal donor lungs before transplantation but is also an excellent platform to apply novel therapeutics. We investigated donor lung immunomodulation using genetically engineered mesenchymal stromal cells with augmented production of human anti-inflammatory hIL-10 (MSCsIL-10). Pig lungs were placed on EVLP for 6 h and randomized to control (n = 7), intravascular delivery of 20 × 106 (n = 5, low dose) or 40 × 106 human MSCs IL-10 (n = 6, high dose). Subsequently, single-lung transplantation was performed, and recipient pigs were monitored for 3 days. hIL-10 secretion was measured during EVLP and after transplantation, and immunological effects were assessed by cytokine profile, T and myeloid cell characterization and mixed lymphocyte reaction. MSCIL-10 therapy rapidly increased hIL-10 during EVLP and resulted in transient hIL-10 elevation after lung transplantation. MSCIL-10 delivery did not affect lung function but was associated with dose-related immunomodulatory effects, with the low dose resulting in a beneficial decrease in apoptosis and lower macrophage activation, but the high MSCIL-10 dose resulting in inflammation and cytotoxic CD8+ T cell activation. MSCIL-10 therapy during EVLP results in a rapid and transient perioperative hIL-10 increase and has a therapeutic window for its immunomodulatory effects.

Ferroptosis, characterized by iron-dependent lipid reactive oxygen species (ROS) accumulation, plays a pivotal role in cisplatin-induced ototoxicity. Existing research has suggested that in cisplatin-mediated damage to auditory cells and hearing loss, ferroptosis is partially implicated. 4-Octyl itaconate (4-OI), derived from itaconic acid, effectively permeates cell membranes, showcasing potent anti-inflammatory as well as antioxidant effects in several disease models. Our study aimed to investigate the effect of 4-OI on cisplatin-induced ferroptosis and the underlying molecular mechanisms. The survival rates of HEI-OC1 cells and mice cochlea hair cells were measured by CCK8 and immunofluorescence, respectively. The auditory brainstem response (ABR) audiometry was used to detect changes in hearing thresholds in mice before and after treatment. Levels of ROS were evaluated by DCFH-DA. Real-time PCR quantified inflammatory cytokines TNF-α, IL-6 and IL-1β. Network Pharmacology and RNA sequencing (RNA-seq) analysis of the potential mechanism of 4-OI resistance to cisplatin-induced ferroptosis. The expressions of ferroptosis-related factors (GPX4, SLC7A11 and PTGS2) and important antioxidant factors (NRF2, HO-1, GCLC and NQO1) were tested by real-time PCR, Western blot and immunofluorescence. Results demonstrated cisplatin-induced significant ROS and inflammatory factor release, reduced NRF2 expression, hindered nuclear translocation and activated ferroptosis. Pretreatment with 4-OI exhibited anti-inflammatory and antioxidant effects, along with resistance to ferroptosis, ultimately mitigating cisplatin-induced cell loss. In the present study, we show that 4-OI inhibits cisplatin-induced ferroptosis possibly through activation of the NRF2/HO-1 signalling pathway, thereby exerting a protective effect against cisplatin-induced damage to auditory cells, and providing a new therapeutic strategy for cisplatin-induced hearing loss.

AIMS:FAM134B, the initial endoplasmic reticulum (ER)-phagy receptor identified, facilitates ER-phagy during ER stress. The malfunction of FAM134B has been demonstrated to have a crucial role in the pathological mechanisms of diverse human ailments. However, the role of FAM134B-mediated ER-phagy in ototoxicity, particularly in cisplatin-induced ototoxicity, remains unclear. The present study endeavors to investigate whether FAM134B is expressed in House Ear Institute-Organ of Corti 1 (HEI-OC1) and C57BL/6 murine cochlear hair cells (HCs), and to explore its potential function in cisplatin-mediated ototoxicity, with the aim of discovering new insights that can mitigate or forestall the irreversible adverse effect of cisplatin.METHODS:Immunofluorescence (IF) staining was used to test the expression pattern of FAM134B, levels of C/EBP-homologous protein (CHOP), autophagy, and co-localization ratio of lysosomes and ER. Western blotting was employed to measure changes in expression levels of FAM134B, LC3B, ER stress-related proteins, LAMP1 and apoptotic mediators. Cell apoptosis was examined using transferase dUTP nick end labeling (TUNEL) assay and flow cytometry.RESULTS:In the present investigation, it was observed that FAM134B exhibited a diffuse expression pattern in the cytoplasm and nuclei of control HEI-OC1 cells. Following cisplatin administration, FAM134B was found to accumulate and form distinct dots around the nuclei, concomitant with increased levels of ER-phagy, ER stress, unfolded protein response (UPR), and cell apoptosis. Additionally, knockdown of FAM134B resulted in reduced ER-phagy, mitigated ER stress and UPR, and decreased apoptotic activity in HEI-OC1 cells following cisplatin exposure.CONCLUSIONS:Collectively, the findings of this study demonstrate that FAM134B-mediated ER-phagy enhances the susceptibility of HCs to ER stress and apoptosis in response to cisplatin-induced stress. This suggests a sequential progression of ER-phagy, ER stress and apoptosis following cisplatin stimulus, and implies the potential therapeutic benefit of inhibiting of FAM134B-mediated ER-phagy in the prevention of cisplatin-related ototoxicity.

The diagnosis and recognition of male osteoporosis is still insufficient. The male patients with low-energy fracture were less evaluated and treated for osteoporosis. The risk of osteoporotic fracture in elderly men may be between 13% and 25%. With the extension of men’s life, the risk of fracture increases, which makes the management of men with fracture risk challenging. A consensus is emerging on targeted treatment guidelines for male osteoporosis, especially for elderly patients with high fracture risk. This paper aims to seek evidence through literature review, and put forward the most effective clinical strategies and schemes for the diagnosis and treatment of senile osteoporosis, so as to provide some theoretical guidance for clinical practice.

Objective: Diagnosing lung injury is a challenge in lung transplantation. It has been unclear if a single biopsy specimen is truly representative of the entire organ. Our objective was to investigate lung inflammatory biomarkers using human lung tissue biopsies and ex vivo lung perfusion perfusate.Methods: Eight human donor lungs declined for transplantation were air inflated, flash frozen, and partitioned from apex to base. Biopsies were then sampled throughout the lung. Perfusate was sampled from 4 lung lobes in 8 additional donor lungs subjected to ex vivo lung perfusion. The levels of interleukin-6, interleukin-8, interleukin-10, and interleukin-1b were measured using quantitative reverse transcription polymerase chain reaction from lung biopsies and enzyme-linked immunosorbent assay from ex vivo lung perfusion perfusate.Results: The median intra-biopsy equal-variance P value was .50 for messenger RNA biomarkers in tissue biopsies. The median intra-biopsy coefficient of variance was 18%. In donors with no apparent focal injuries, the biopsies in each donor showed no difference in various lung slices, with a coefficient of variance of 20%. The exception was biopsies from the lingula and injured focal areas that demonstrated larger differences. Cytokines in ex vivo lung perfusion perfusate showed minimal variation among different lobes (coefficient of variance 1/4 4.9%).Conclusions: Cytokine gene expression in lung biopsies was consistent, and the biopsy analysis reflects the whole lung, except when specimens were collected from the lingula or an area of focal injury. Ex vivo lung perfusion perfusate also provides a representative measurement of lung inflammation from the draining lobe. These results will reassure clinicians that a lung biopsy or an ex vivo lung perfusion perfusate sample can be used to inform donor lung selection. (J Thorac Cardiovasc Surg 2023;166:1520-8)

Hearing loss is currently one of the most prevalent sensory disorders worldwide. Because both the blood-labyrinth barrier and the limited blood circulation in the inner ear restrain the effective delivery of most drugs to the inner ear tissues, current treatments for hearing loss are limited to mainly medication, hearing devices and cochlear surgery for therapeutic purposes, whereas treatments lack a noninvasive targeted drug-delivery system. With the continuously rapid development of new nanomaterials, the nanodelivery systems are expected to provide a potentially effective method of clinical treatment for hearing loss. This paper reviews the advantages and disadvantages of the commonly used drug-delivery methods and novel nanomaterials for inner ears as well as advancements in the targeted treatment of hearing loss.

Background. Alzheimer's disease (AD) is closely related to aging, showing an increasing incidence rate for years. As one of the main organs involved in AD, hippocampus has been extensively studied due to its association with many human diseases. However, little knowledge is known on its association with primary ciliary dyskinesia (PCD).Material and Methods. The microarray data of hippocampus on AD were retrieved from the Gene Expression Omnibus (GEO) database to construct the co-expression network by weighted gene co-expression network analysis (WGCNA). The gene network modules associated with AD screened with the common genes were further annotated based on Gene Ontology (GO) database and enriched based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. The protein-protein interaction (PPI) network was constructed based on STRING database to identify the hub genes in the network.Results. Genes involved in PCD were identified in the hippocampus of AD patients. Functional analysis revealed that these genes were mainly enriched in ciliary tissue, ciliary assembly, axoneme assembly, ciliary movement, microtubule based process, microtubule based movement, organelle assembly, axoneme dynamin complex, cell projection tissue, and microtubule cytoskeleton tissue. A total of 20 central genes, e.g.,DYNLRB2, ZMYND10, DRC1, DNAH5, WDR16, TTC25, and ARMC4 were identified as hub genes related to PCD in hippocampus of AD patients.Conclusion. Our study demonstrated that AD and PCD have shared metabolic pathways. These common pathways provide novel evidence for further investigation of the pathophysiological mechanism and the hub genes suggest new therapeutic targets for the diagnosis and treatment of AD and PCD.Subjects Bioinformatics, Cell Biology, Molecular Biology, Neurology

Background:Nonsyndromic low-frequency sensorineural hearing loss (LFSNHL) is an uncommon form of hearing loss (HL) that typically affects frequencies at 2000 Hz and below. Heterozygous variants in the WFS1 gene at the DFNA6/14/38 locus are considered a common cause of LFSNHL. To date, 34 different pathogenic genetic variants have been reported to cause LFSNHL with seven of these variants identified in the Chinese population. However, limited reports are available on the association between WFS1 gene and LFSNHL. Here, we report a five-generation Chinese family with an autosomal dominant inheritance pattern of postlingual and progressive LFSNHL.Methods:Routine clinical and audiological examinations were performed on 16 affected and 7 healthy members in this family. The targeted next-generation sequencing of 127 known deafness genes was performed to identify variants in affected individuals. Sanger sequencing were further employed to confirm the pathogenic variant identified.Results:A novel heterozygous pathogenic genetic variant c.2530G > T (p.Ala844Ser) was identified in the WFS1 gene in all patients of this family. The mutated Ala residue is evolutionarily conserved and cosegregated with HL. The variant was predicted to be deleterious by MutationTaster, PolyPhen-2, LRT, and Fathmm software. Conservation analysis and 3D protein structure model indicated that the variant caused a structural change in the protein.Conclusions:Our present study identifies a novel heterozygous WFS1 variant associated with LFSNHL in a Chinese family.

AIM:To explore the relationship between C-reactive protein (CRP) and mortality in critically ill patients with acute kidney injury (AKI). MATERIALS AND METHODS:A total of 580 patients diagnosed with AKI within 48 hours of ICU admission between September 2017 and August 2019 were enrolled. Patients were followed for all-cause mortality in-hospital and then up to 2 years after discharge. We performed two multivariate regression analysis to assess the association between CRP and mortality, and conducted stratified analysis to assess whether the effect of the CRP differed across subgroups. RESULTS:According to initial CRP quartiles, patients were divided into 4 groups (quartile 1, CRP ≤ 2.87 mg/L; quartile 2, CRP: 2.87 - 25.95 mg/L; quartile 3, CRP: 25.95 - 111.51 mg/L; quartile 4, CRP > 111.51 mg/L). Patients with high CRP levels have higher APACHE-II score, longer length of stay in the ICU, and higher mortality. In multivariate regression analysis, high CRP was associated with the increased risk of in-hospital mortality after adjusting for age, gender, surgical grade, heart rate, serum potassium, serum chloride, coronary heart disease, and atherosclerotic cerebral infarction (quartile 4 vs. quartile 1, OR: 3.810, 95% CI: 2.081 - 6.973). For 2-year mortality, the increased trend was still significant with the OR (95% CI) of the quartile 4 group of 5.117 (2.678 - 9.780) after adjusting for confounders. Subgroup analyses detected in each group showed that the in-hospital and 2-year risk of mortality increased with higher CRP levels. CONCLUSION:Higher CRP level was associated with the increased risk of mortality in critically ill patients with AKI.

Background: The 4-octyl itaconate (OI) is a type of cell-permeable itaconate derivative. Studies have shown that with an anti-fibrotic effect in systemic sclerosis, the OI also affects osteoclast differentiation. The aim of this study was to explore the molecular mechanisms underlying the effects of OI on myoblast differentiation by RNA-seq analysis.Methods: Myoblast proliferation, differentiation, and muscle regulatory factors were examined in C2C12 myo-blasts treated with OI of various concentrations (2.5, 10, 25, 50, and 100 mu mol/L). Cells were treated with the PI3K-Akt activator IGF-1 to explore the role of the PI3K-Akt pathway in OI inhibition of myogenic differentiation. The regulatory mechanisms of OI in myogenesis were further investigated by RNA-seq and subsequent gene ontology (GO), kyoto encyclopedia of genes and genomes (KEGG) and, gene set enrichment analysis (GSEA).Results: OI of various concentrations did not show any effect during cell proliferation. During differentiation, OI inhibited the expressions of the marker of mature myotubes myosin heavy chain (MHC) and myogenin in a dose -dependent manner. OI inhibited muscle differentiation by affecting MyoD-regulated activity through inhibition of AKT1 phosphorylation. The results of the KEGG enrichment analysis and GSEA showed that OI affected multiple metabolic pathways during myogenic differentiation, including PI3K-Akt signaling, calcium signaling, and PPAR signaling.Conclusions: Our study broadens the understanding of the OI inhibition of myogenic differentiation. OI plays its functions by targeting multiple molecules and pathways, providing novel insights into the understanding of the overall effect of OI.

Aims: The present study was aimed to explore the possible mechanism(s) underlying the action of cisplatin on auditory cells of mice in vitro, with special attention given to the dynamic variation in calcium homeostasis and responding channels. Methods: The apoptosis of auditory cells was tested by flow cytometry and TUNEL staining. The expressions of inositol 1,4,5-trisphosphate receptors (IP3R), voltage-dependent anion channel 1 (VDAC1), phosphorylated protein kinase R-like ER kinase (p-PERK), activating transcription factor 6 (ATF6), caspase-12, bcl-2, bax, cleaved caspase-9, cleaved caspase-3, beclin-1 and light chain 3p (LC3B) were measured by immunofluorescence or Western blotting. The calcium variations in subcellular structures were evaluated by Rhod-2 AM and MagFluo-4 AM staining. The colocalization ratio between IP3R and beclin-1 was determined by immunocytochemistry. Results: We found that cisplatin exposure induced the apoptosis of HEI-OC1 cells and hair cells (HCs) in a caspase3 dependent manner. This apoptotic process was attributed to the activation of endoplasmic reticulum (ER) stress and mitochondrial pathway and, meanwhile, accompanied by variation in calcium homeostasis and responding channels. Interestingly, we also observed that IP3R might dissociate from beclin-1 to motivate autophagy under the cisplatin insult. Conclusions: Overall, the findings from this work indicate that cisplatin leads to auditory cell damage of mice in vitro, which is closely relevant to dynamic variation in calcium homeostasis and responding channels in subcellular structure.

Kretschmer et al. reported in this journal that SARS-CoV-2 lateral flow tests (LFT) for rapid antigen testing resulted in an unnecessary burden in Germany with statistics on rapid antigen testing conducted in Cologne, in May 2021.1Kretschmer A. Kossow A. Grüne B. Schildgen O. Mathes T. Schildgen V. False positive rapid antigen tests for SARS-CoV-2 in the real-world and their economic burden.J Infect. 2021; (S0163-4453(21)00402-3)https://doi.org/10.1016/j.jinf.2021.08.020Google Scholar In this study, they used the PCR test to determine the false positive rate of the antigen test. The results showed that the total false positive rate of rapid antigen detection was as high as 46.28%. Andreas and colleagues concluded that "From a pure laboratory and diagnostic point of view it has to be concluded that the usage of an antigen testing strategy with a false detecting rate of about 50% is unacceptable."1Kretschmer A. Kossow A. Grüne B. Schildgen O. Mathes T. Schildgen V. False positive rapid antigen tests for SARS-CoV-2 in the real-world and their economic burden.J Infect. 2021; (S0163-4453(21)00402-3)https://doi.org/10.1016/j.jinf.2021.08.020Google Scholar To obtain the insights into the economic burden of rapid antigen testing, they calculated the direct costs arising from the performed tests and the indirect costs arising from the subsequent quarantine. The authors further concluded that "the use of rapid antigen testing only appears appropriate in high prevalence/incidence situations, because a (too) low prevalence may increase the risk of false-positive results leading to unnecessary quarantine and high economic burden."1Kretschmer A. Kossow A. Grüne B. Schildgen O. Mathes T. Schildgen V. False positive rapid antigen tests for SARS-CoV-2 in the real-world and their economic burden.J Infect. 2021; (S0163-4453(21)00402-3)https://doi.org/10.1016/j.jinf.2021.08.020Google Scholar Currently, the diagnosis of COVID-19 is mainly based on testing SARS-CoV-2 RNA load using quantitative real-time polymerase chain reaction (qRT-PCR).2Hadi J. Dunowska M. Wu S. Brightwell G. Control measures for SARS-CoV-2: a review on light-based inactivation of single-stranded RNA viruses.Pathogens. 2020; 9: 737https://doi.org/10.3390/pathogens9090737Google Scholar Although the detection method using qRT-PCR has been considered as the benchmark among the currently available diagnostic approaches, strategies to rapidly scale up the testing for SARS-CoV-2 must be considered for future diagnostic testing, particularly dealing with the circumstances encountered worldwide in the past year. Nucleic acid testing usually requires considerable laboratory equipment and staff, and test results may take several days to become available to the physician.3Li K. Huang B. Wu M. Zhong A. Li L. Cai Y. et al.Dynamic changes in anti-SARS-CoV-2 antibodies during SARS-CoV-2 infection and recovery from COVID-19.Nat Commun. 2020; 11: 6044https://doi.org/10.1038/s41467-020-19943-yGoogle Scholar Therefore, rapid antigen testing is a crucial supplement to nucleic acid testing for COVID-19 management with the test results obtained rather quickly.4Augustine R. Das S. Hasan A.S.A. Abdul Salam S. Augustine P. et al.Rapid antibody-based COVID-19 mass surveillance: relevance, challenges, and prospects in a pandemic and post-pandemic world.J Clin Med. 2020; 9: 3372https://doi.org/10.3390/jcm9103372Google Scholar However, taking the experience of Kretschmer et al.1Kretschmer A. Kossow A. Grüne B. Schildgen O. Mathes T. Schildgen V. False positive rapid antigen tests for SARS-CoV-2 in the real-world and their economic burden.J Infect. 2021; (S0163-4453(21)00402-3)https://doi.org/10.1016/j.jinf.2021.08.020Google Scholar into account, with the rapid antigen testing leading to unnecessary quarantine and high economic burden, the novel rapid and inexpensive testing methods are therefore valuable. To deal with these difficulties in detecting the SARS-CoV-2, the Integrated Mobile Container PCR Laboratory (IMCPL) unit was developed in China, which was a standard PCR laboratory established inside a walk-in container and can be quickly set up at any convenient locations to carry out the large-scale diagnostic testing. Two main characteristics of the IMCPL make this medical unit more suitable for diagnostic detection than the traditional laboratories in the event of major public health emergencies. First, the IMCPL unit is extremely sturdy and mobile (Fig. 1). The square rectangle cabin is 17.5 m in length, 3 m width, and 3 m height, built using steel plates of 3 mm thickness with each part further reinforced to ensure the safety during transportation. In order to be deployed rapidly, the IMCPL unit can be conveniently carried and transported by using a logistics 17.5 m semi-trailer (Fig. 1A), enabling the long-distance and cross-regional laboratory support facilitated in a short time. Second, the installation requirements of the IMCPL unit are minimal. The IMCPL unit can be installed quickly at any convenient sites under most circumstances in less than 2 h with the availability of both water and electricity. To assess the operating efficiency of the IMCPL unit, we carried out the tests in the IMCPL unit at the Shandong Provincial Hospital in China. The processing speed of the laboratory completing the sample testing depends largely on the proficiency of the personnel members and the operational efficiency of the supporting equipment.5Safiabadi Tali S.H. LeBlanc J.J. Sadiq Z. Oyewunmi O.D. Camargo C. Nikpour B. et al.Tools and techniques for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)/COVID-19 detection.Clin Microbiol Rev. 2021; 34 (e00228-20)https://doi.org/10.1128/CMR.00228-20Google Scholar In our study, several skilled medical laboratory technicians performed individually a load test on the supporting equipment installed in the IMCPL unit, including the hot thermostat pan (used to inactivate viruses), nucleic acid extractor, and the fluorescent quantitative PCR analyzer (Fig. 2A-C). The experimental procedure was performed according to the manual of the kits. The results showed that it took a skilled technician ∼35 min to complete the sample inactivation, 20–40 min to finish the nucleic acid extraction, and 90–120 min to run the nucleic acid amplification (Fig. 2D). These results showed evidently that the operation efficiency on the equipment was greatly improved in the IMCPL unit in comparison to normal process performed in a regular molecular laboratory. Overall, it took a skilled technician 135–195 min to complete the detection of viral nucleic acid in a batch of 96 samples in the IMCPL unit. In order to maintain the high work efficiency, we organized four working shifts every day with a group of staff members replaced and the instruments disinfected every 6 h. The results showed that an IMCPL unit equipped with 12 PCR analyzers completed the nucleic acid detection of ∼8000–10,000 clinical samples within 24 h (Fig. 2E), demonstrating evidently the enhanced SARS-CoV-2 detection capability of the IMCPL unit in a high-throughput manner. It is worth noting that under the condition of low incidence, IMCPL can be used as an independent laboratory configured by the hospital to complete the daily testing work. The Shandong Provincial Hospital began to operate the IMCPL unit on January 7, 2021, with a total of 132,027 samples processed by August 14, 2021, significantly alleviating the workload pressure on the clinical laboratory at the hospital. The large-scale SARS-CoV-2 screening is necessary to prevent the spread of the disease COVID-19, to help government make relevant public health policies, and to test people and their close contacts with a history of travel or residence in the epidemic areas. The IMCPL unit can be used as a reserve force for the health and epidemic prevention programs, ultimately reducing the unnecessary high positive rate and economic burden caused by the application of rapid antigen testing. Under the current unpredictable situations in many countries globally, the IMCPL unit could be used in combination with the local medical laboratories to eliminate the blind spots in the areas with epidemic. The authors declare that no competing interests exist related to this submission. This work was supported by the National Natural Science Foundation of China (Nos. 81670942) and Special Funds for Taishan Scholar Project (Nos.tsqn202103180).

Prostate cancer (PC) is a common tumor in men, and the incidence rate is high worldwide. Exosomes are nanosized vesicles released by all types of cells into multiple biological fluid types. These vesicles contribute to intercellular communication by delivering both nucleic acids and proteins to recipient cells. In recent years, many studies have explored the mechanisms by which exosomes mediate the epithelial-mesenchymal transition, angiogenesis, tumor microenvironment establishment, and drug resistance acquisition in PC, and the mechanisms that have been identified and the molecules involved have provided new perspectives for the possible discovery of novel diagnostic markers in PC. Furthermore, the excellent biophysical properties of exosomes, such as their high stability, high biocompatibility and ability to cross biological barriers, have made exosomes promising candidates for use in novel targeted drug delivery system development. In this review, we summarize the roles of exosomes in the growth and signal transmission in PC and show the promising future of exosome contributions to PC diagnostics and treatment.

As a chronic metabolic disease caused by disorders of purine metabolism, gout has shown increasing incidence rate worldwide. Considering that gout is not easily treated and cured, further studies are explored to prevent gout development through diet modification. Both β-carotin and green tea powder are rich in dietary fiber, which helps maintain the balance of gut microbiota in humans. The aim of this study was to investigate the effects of β-carotin and green tea powder diet on the prevention of gouty arthritis in relation to the bacterial structure of gut microbiota in mice. We successfully induced gouty arthritis in C57BL/6 mice by injecting monosodium urate (MSU) crystals and feeding high-fat diet (HFD), and further investigated the effects of additional β-carotin and green tea powder in the diets of mice on the prevention of gouty arthritis in mice. Our results showed that diet of β-carotin and green tea powder reduced the joint swelling and pain in mice with gout, reduced the levels of serum uric acid (UA) and three types of pro-inflammatory cytokines, i.e., interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α), improved the gut microbiota profile, and reduced the metabolic levels of purines and pyrimidines. In conclusion, our study provided evidence to support the application of β-carotin and green tea powder diet as a dietary adjustment method to prevent and treat gouty arthritis.

Background: Auditory neuropathy (AN) is a specific type of hearing loss characterized by impaired language comprehension. Apoptosis inducing factor mitochondrion associated 1 ( AIFM1 ) is the most common gene associated with late-onset AN. In this study, we aimed to screen the pathogenic variant of AIFM1 in a Chinese family with AN and to explore the molecular mechanism underlying the function of such variant in the development of AN. Methods: One patient with AN and eight unaffected individuals from a Chinese family were enrolled in this study. A comprehensive clinical evaluation was performed on all participants. A targeted next-generation sequencing (NGS) analysis of a total of 406 known deafness genes was performed to screen the potential pathogenic variants in the proband. Sanger sequencing was used to confirm the variants identified in all participants. The pathogenicity of variant was predicted by bioinformatics analysis. Immunofluorescence and Western blot analyses were performed to evaluate the subcellular distribution and expression of the wild type (WT) and mutant AIFM1 proteins. Cell apoptosis was evaluated based on the TUNEL analyses. Results: Based on the clinical evaluations, the proband in this family was diagnosed with AN. The results of NGS and Sanger sequencing showed that a novel missense mutation of AIFM1 , i.e., c.1367A > G (p. D456G), was identified in this family. Bioinformatics analysis indicated that this variant was pathogenic. Functional analysis showed that in comparison with the WT, the mutation c.1367A > G of AIFM1 showed no effect on its subcellular localization and the ability to induce apoptosis, but changed its protein expression level. Conclusion: A novel variant of AIFM1 was identified for the first time, which was probably the genetic cause of AN in a Chinese family with AN.

Small molecule drug intervention for chondrocytes is a valuable method for the treatment of osteoarthritis (OA). The 4‐octyl itaconate (OI) is a cellular derivative of itaconate with sound cell permeability and transformation rate. We attempted to confirm the protective role of OI in chondrocytes and its regulatory mechanism. We used lipopolysaccharide (LPS) to induce chondrocyte inflammation injury. After the OI treatment, the secretion and mRNA expression of Il ‐ 6 , Il ‐ 10 , Mcp ‐ 1 and Tnf ‐ α were detected by ELISA and qPCR. The protective effect of OI on articular cartilage was further verified in surgical destabilization of the medial meniscus model of OA. Cell death and apoptosis were evaluated based on CCK8, LDH, Typan blue staining, Annexin V and TUNEL analyses. The small interfering RNAs were used to knockout the Nrf2 gene of chondrocytes to verify the OI‐mediated Nrf2 signalling pathway. The results revealed that OI protects cells from LPS‐induced inflammatory injury and attenuates cell death and apoptosis induced by LPS. Similar protective effects were also observed on articular cartilage in mice. The OI activated Nrf2 signalling pathway and promoted the stable expression and translocation of Nrf2 into the nucleus. When the Nrf2 signalling pathway was blocked, the protective effect of OI was significantly counteracted in chondrocytes and a mouse arthritis model. Both itaconate and its derivative (i.e., OI) showed important medical effects in the treatment of OA.

Additional file 10: Table S6. Gene Interaction Heatmap data.

Objective: The zebrafish is an excellent model for studying gene function in auditory system development. Pou4f3 plays an important role in mouse hair cell formation. Here, we constructed a pou4f3 -knockout Tg(Brn3c:GFP) zebrafish to provide an efficient fluorescence-visualized model for studying the molecular mechanisms of ear development. Methods: Cas9/single guide RNAs targeting exon 2 of pou4f3 were designed and injected into one-cell stage zebrafish embryos (G0 generation). The G0 generation were crossed with Tg(Brn3c:GFP) zebrafish to obtain pou4f3- mutant Tg(Brn3c:GFP) zebrafish. The targeting efficiency was detected by polymerase chain reaction amplification and Sanger sequencing. Zebrafish hair cells were observed by laser scanning confocal microscopy in vivo . The morphology of the otoliths and semicircular canals were analyzed. All animal experiments were approved by the Animal Care and Use Committee of Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University (approval No. 2016-KY-040) on March 3, 2016. Results: The pou4f3 -mutant Tg(Brn3c:GFP) zebrafish line was successfully established. Fluorescence observation suggested that hair cell development was delayed in pou4f3 -knockout zebrafish. Knockout of pou4f3 also induced defects in the otoliths and semicircular canals and impaired ear function in zebrafish. Conclusion: A CRISPR/Cas9-mediated pou4f3 mutant Tg(Brn3c:GFP) zebrafish model was established for the first time to demonstrate the essential role of pou4f3 in zebrafish ear development. Our study provides a highly efficient method for the establishment of a visualized model of gene knockout zebrafish and has the potential to allow high-throughput drug screening to explore therapeutics for related diseases.

The small muscle protein, x-linked (SMPX) encodes a small protein containing 88 amino acids. Malfunction of this protein can cause a sex-linked non-syndromic hearing loss, named X-linked deafness 4 (DFNX4). Herein, we reported a point mutation and a frameshift mutation in two Chinese families who developed gradual hearing loss with age. To explore the impaired sites in the hearing system and the mechanism of DFNX4, we established and validated an Smpx null mouse model using CRISPR-Cas9. By analyzing auditory brainstem response (ABR), male Smpx null mice showed a progressive hearing loss starting from high frequency at the 3rd month. Hearing loss in female mice was milder and occurred later compared to male mice, which was very similar to human beings. Through morphological analyses of mice cochleas, we found the hair cell bundles progressively degenerated from the shortest row. Cellular edema occurred at the end phase of stereocilia degeneration, followed by cell death. By transfecting exogenous fluorescent Smpx into living hair cells, Smpx was observed to be expressed in stereocilia. Through noise exposure, it was shown that Smpx might participate in maintaining hair cell bundles. This Smpx knock-out mouse might be used as a suitable model to explore the pathology of DFNX4.