BackgroundPatients with prolonged disorders of consciousness (pDoC) present significant clinical challenges, directly influencing awakening strategies and the allocation of healthcare resources. This study aimed to evaluate serum sex hormone levels in pDoC patients and assess their potential for predicting clinical outcomes.MethodsThis retrospective study initially screened 142 patients with prolonged disorders of consciousness (pDoC) admitted between January 2023 and January 2025. After applying exclusion criteria, a final cohort of 119 patients (77 males and 42 females) was included for analysis. Within 1 week of enrollment, the Chinese Coma Recovery Scale-Revised (CRS-R) score and serum sex hormone levels were assessed. Outcomes were evaluated at 6 months using the Glasgow Outcome Scale–Extended (GOS-E) and classified as good outcome(GOSE score≥4) or poor outcome(GOSE score <4). Statistical analyses were performed to explore the association between serum sex hormone levels and GOSE scores to identify potential prognostic markers.ResultsAmong male patients, the good outcome group showed higher testosterone levels (p = 0.02) and a trend toward lower prolactin levels (p = 0.06) compared to the poor outcome group. In female patients, the good outcome group had lower levels of prolactin, follicle-stimulating hormone, and luteinizing hormone (p < 0.05). However, after Bonferroni correction for multiple comparisons, none of these associations remained statistically significant. In multivariate logistic regression adjusting for age, disease duration, and etiology, log-transformed testosterone showed a consistent association with favorable outcomes in males (odds ratio, OR = 2.97, 95% CI: 1.29–7.70, p = 0.02), though this did not survive Bonferroni correction (P_bonf = 0.10). Prolactin and other sex hormones were not independent predictors in either sex. Receiver operating characteristic (ROC) analysis demonstrated that testosterone alone yielded an area under the curve (AUC) of 0.656 (95% CI: 0.532–0.781), with minimal improvement after etiology adjustment (AUC = 0.691, 95% CI: 0.573–0.809; p = 0.45).ConclusionSerum testosterone showed exploratory, hypothesis-generating associations with favorable outcomes in male pDoC patients. However, these findings did not survive correction for multiple testing, and the predictive performance was modest. Sex hormone levels did not demonstrate clear prognostic value in female patients. These preliminary observations underscore the need for larger, prospective studies with standardized endocrine protocols to validate the potential role of sex hormones as prognostic biomarkers in pDoC.

This study aimed to explore the value of sympathetic skin response (SSR) for identifying paroxysmal sympathetic hyperactivity (PSH) in patients with prolonged disorders of consciousness (PDOC). This retrospective observational study included 124 consecutive patients with PDOC from March 2022 to March 2024. On the basis of the PSH Assessment Method (PSH-AM), the patients were classified into the PSH-positive (PSH + , n = 43) and PSH-negative (PSH − , n = 81) groups. Among them, 75 patients (PSH − , n = 44; PSH + , n = 31) with elicitable SSR were included in the final analysis. There were no significant differences between groups in the SSR elicitation rate (P = 0.08) or in SSR latency (P > 0.05). By contrast, both SSR amplitude and SSR amplitude difference were significantly elevated in the PSH + group compared with the PSH- group (P < 0.001). In patients with elicitable SSR (n = 75), multivariable logistic regression analysis including age, CRS-R score, right SSR amplitude, left SSR amplitude, and SSR amplitude difference simultaneously (with no significant multicollinearity) demonstrated that only the SSR amplitude difference was independently associated with PSH (OR per 0.1 mV = 2.08; 95

Objective This study aimed to investigate the characteristics of the blink reflex and its prognostic value for consciousness improvement in patients with prolonged disorders of consciousness (DOC).Design Prospective case-control study.Setting This study was conducted in a local hospital between March 2022 and March 2023.Participants Patients in a vegetative state/unresponsive wakefulness syndrome or in a minimally conscious state were enrolled within 3 months from their brain injury.Primary and secondary outcome measures The early component (R1), ipsilateral late component (iR2) and contralateral late component (cR2) responses at baseline were recorded using electromyography. The patients’ clinical diagnosis and the best Coma Recovery Scale-Revised (CRS-R) total score were assessed based on Chinese CRS-R evaluations.Outcome definition At the 6-month follow-up, patients were categorised as improved or non-improved based on CRS-R score changes (improved (transition to a higher consciousness state); non-improved (worsened condition, static or death)).Results A total of 58 DOC patients were included in this study. Of the 58 DOC patients, 32 were classified as the improved group and 26 as the non-improved group. In the improved group, R2 responses were elicited in 30 patients, while only 16 patients in the non-improved group had elicited R2 responses. The non-improved group exhibited significantly lower R2 mean amplitudes (iR2 (105.08 µV vs 173.25 µV, p=0.01); cR2 (55.15 µV vs 114.03 µV, p=0.01)) and longer mean latencies (iR2 (41.08±6.72 ms vs 37.77±3.94 ms, p=0.03); cR2 (41.32±6.28 ms vs 37.48±4.07 ms, p=0.01)) compared with the improved group. The result demonstrated that the iR2 mean amplitude (OR=1.01, area under the curve (AUC)=0.78 (95% CI 0.63 to 0.93), sensitivity=78.12%, specificity=83.33%, p=0.02) and cR2 mean amplitude (OR=1.02, AUC=0.76 (95% CI 0.62 to 0.90), sensitivity=81.25%, specificity=72.22%, p=0.02) were significant predictors of consciousness improvement. Meanwhile, Pearson correlation analysis revealed that iR2 mean amplitude (r=0.42, p=0.003) and cR2 mean amplitude (r=0.53, p=0.001) significantly correlated with CRS-R score at baseline.Conclusion The R2 amplitude in patients with prolonged DOC may serve as a prognostic indicator for consciousness improvement.

To evaluate the prognostic value of somatosensory evoked potentials (SEPs) in predicting functional outcomes in patients with prolonged disorders of consciousness (PDOC) secondary to severe supratentorial intracerebral hemorrhage (ICH). From August 2023 to August 2024, 77 patients with PDOC secondary to supratentorial intracerebral hemorrhage were prospectively enrolled from Critical Care Rehabilitation Department of Nanjing Jiangning Hospital. Within one week of enrollment, all patients were assessed using the Chinese Coma Recovery Scale-Revised (CRS-R) score, short-latency somatosensory evoked potentials (SLSEPs), and middle-latency somatosensory evoked potentials (MLSEPs). Patients were followed for 6 months, with outcomes classified using the Extended Glasgow Outcome Scale (GOSE). Based on GOSE scores, patients were categorized into good outcome groups (GOSE score ≥ 4) and poor outcome groups(GOSE score < 4). A total of 77 PDOC patients were included in the study. After 6 months of follow-up, 44 patients (57.14

Muscle atrophy in pathological or diseased muscles arises from an imbalance between protein synthesis and degradation. Elevated levels of interleukin-6 (IL-6) are a hallmark of ischemic stroke and have been associated with muscle atrophy in certain pathological contexts. However, the mechanisms by which IL-6 induces muscle atrophy in the context of stroke remain unclear. To investigate these effects, we used a rat model of middle cerebral artery occlusion (MCAO) and an in vitro model with the C2C12 cell line to uncover potential molecular mechanisms underlying IL-6-induced muscle atrophy. Our findings revealed elevated protein and serum levels of IL-6, along with increased markers of muscle atrophy, in MCAO rats compared to sham controls. We also observed overactivation of protein ubiquitination pathways and downregulation of muscle regeneration markers in MCAO rats. Further analysis indicated that IL-6 contributes to increased muscle protein ubiquitination. Inhibition of IL-6 signaling led to a significant reduction in infarct size and improved neurological deficit scores. Targeting the IL-6/IL-6R signaling pathway presents a promising therapeutic approach to mitigate muscle atrophy in individuals affected by ischemic stroke.

BACKGROUND:The recurrence and metastasis of oesophageal squamous cell cancer (ESCC) following radiation therapy are major treatment challenges. Cancer-associated fibroblasts (CAFs) are key in the ESCC microenvironment, yet their role in post-radiation recurrence remains unclear. MATERIALS AND METHODS:KYSE150 ESCC cells were co-implanted with non-irradiated (0 Gy) or irradiated (8 Gy) CAFs in nude mice. CAF-derived extracellular vesicles (EVs) were isolated via differential centrifugation and analysed by electron microscopy and immunoblotting. Transwell assays evaluated EVs' effects on ESCC cell migration and invasion in vitro. RNA sequencing identified differentially expressed microRNAs, and functional experiments verified the role of miR-193a-3p. Plasma samples from 32 ESCC patients and tissue samples from 76 ESCC patients were analysed for miR-193a-3p expression. RESULTS:Irradiated CAFs promoted the lung metastasis of ESCC cells in vivo, and their EVs enhanced ESCC cell invasion, migration and metastasis. Elevated miR-193a-3p levels in EVs from irradiated CAFs increased miR-193a-3p expression in ESCC cells. This effect was effectively attenuated by RNase and Triton X-100 (degrading microRNAs encapsulated in EVs), or GW4869 (inhibiting EVs biogenesis and secretion)-indicating that miR-193a-3p functions in an EV-dependent manner. Knockdown of miR-193a-3p diminished the invasion, migration and epithelial-mesenchymal transition (EMT)-promoting activities of CAF-derived EVs. Luciferase assays confirmed PTEN as a target of miR-193a-3p; miR-193a-3p overexpression decreased PTEN and increased p-Akt expression. In vivo, coinjection of miR-193a-3p-knockdown CAFs with KYSE150 ESCC cells resulted in smaller tumours, fewer lung metastases, increased PTEN and E-cadherin, and decreased p-Akt and Snail expression. Clinically, radiation increased plasma exosomal miR-193a-3p levels, and high miR-193a-3p expression was correlated with shorter survival, identifying miR-193a-3p as an independent predictor of poor prognosis in ESCC patients. CONCLUSION:EVs from irradiated CAFs promote ESCC metastasis via the miR-193a-3p-mediated PTEN/Akt signalling pathway. Targeting this EVs-mediated interaction represents a promising strategy for improving ESCC radiotherapy outcomes. KEY POINTS:The poor prognosis of oesophageal squamous cell carcinoma (ESCC) is largely driven by recurrence and metastasis following radiation therapy. Irradiated cancer-associated fibroblasts (CAFs) drive ESCC recurrence and metastasis through extracellular vesicles (EVs), highlighting their critical role in the post-radiation tumor microenvironment. CAF-derived EVs deliver miR-193a-3p to ESCC cells, suppressing PTEN and activating Akt signaling, thereby enhancing invasion, migration, epithelialmesenchymal transition (EMT), and metastatic potential. High plasma exosomal miR-193a-3p levels predict poor prognosis in ESCC patients and may guide therapeutic strategies after radiotherapy.

To establish a nomogram prediction model for the patients with prolonged disorders of consciousness (PDOC) caused by brain injury at six months based on behavioral scale scores, neuroelectro-physiological techniques and hypothalamic-pituitary hormone levels. The clinical data of patients with PDOC who were first diagnosed and hospitalized in the Department of Rehabilitation Medicine of The Affiliated Jiangning Hospital of Nanjing Medical University from March 2023 to July 2024 were collected retrospectively. We performed stratified sampling based on etiology and divided into a training set (121 cases) and a validation set (49 cases) in a ratio of 7:3. After a 6-month follow-up, patients were divided into groups with improved consciousness and those without improved consciousness based on changes in CRS-R scores.Clinical behavioral scores, somatosensory evoked potentials, brainstem auditory evoked potentials, and levels of hypothalamic-pituitary hormones were utilized to identify prognostic factors for prolonged disorders of consciousness. Concurrently, a nomogram prediction model was crafted and validated to forecast the prognosis of patients with prolonged disorders of consciousness. Decision curve analysis (DCA) was subsequently employed to appraise the clinical applicability of this predictive model. The comparison of clinical data between the training and validation cohorts revealed no significant statistical disparities (P > 0.05). Within the training cohort of 121 PDOC patients, 63 (52.1

Microglia, the primary resident immune cells in the brain, exhibit two distinct functional states: The detrimental (M1) phenotype and the protective (M2) phenotype. Exosomes, which are released by various cell types, play crucial roles in intercellular communication. While existing studies have shown that exosomes from microglia with different activation states can affect neuronal survival following ischemic stroke, a comprehensive exploration of the differences between these microglial phenotypes is still lacking. In this study, we treated primary microglia with lipopolysaccharide(LPS) or interleukin-4 (IL-4) to induce the M1 or M2 phenotype, respectively, and investigated the characteristics of the resulting exosomes. These microglia-derived exosomes can be internalized by neurons. Specifically, exosomes derived from M1microglia (M1-EXOs) exacerbated ischemia-induced neuronal apoptosis both in vivo and in vitro, while exosomes from M2 microglia (M2-EXOs) exhibited a protective effect. Subsequently, we conducted a quantitative proteomic analysis of M1-EXOsand M2-EXOs, characterizing 1129 proteins. Notably, M1-EXO proteins were primarily associated with inflammatory responses, neutrophil chemotaxis, and complement activation. In contrast, M2-EXO proteins were predominantly involved in protein transport and cellular proliferation. In addition, we analyzed key proteins, includingIL-6, SAA3, CCL5, CCL9, C3, CFB, SRGN, and sphingosine-1-phosphate phosphatase 1,which play central roles in the protein-protein interaction network. Overall, this dataset provides valuable insights into the proteomic profiles of exosomes derived from microglia with distinct phenotypes, enhancing our understanding of the mechanisms underlying microglial involvement in central nervous system diseases.

BACKGROUND:Esophageal squamous cell carcinoma (ESCC) is a prevalent malignant tumor worldwide. Circular RNA (circRNA) is of great value in tumorigenesis progression. However, the mechanism of circFNDC3B in ESCC remains to be clarified.METHODS:Firstly, the circular characteristics of circFNDC3B were evaluated by Actinomycin D and RNase R measurements. The functions of circFNDC3B in ESCC cells were examined by CCK-8, EdU and flow cytometry. Subsequently, the molecular mechanism of circFNDC3B was explained using luciferase reporter gene detection. Finally, we constructed xenograft model to prove the role of circFNDC3B in vivo.RESULTS:Our study revealed that circFNDC3B was more stable than its linear RNA and prominently upregulated in ESCC. Functional findings suggested that silencing of circFNDC3B reduced the proliferation and enhanced apoptosis of ESCC cells in vitro. Meanwhile, knockdown of circFNDC3B attenuated tumor progression in vivo. Next, miR-370-3p/miR-136-5p was discovered to bind circFNDC3B. miR-370-3p/miR-136-5p reversed the promotive effect on cell proliferation and the inhibitory effect on cell apoptosis of circFNDC3B. MYO5A was a downstream target of miR-370-3p/miR-136-5p. CircFNDC3B served as a sponge for miR-370-3p/miR-136-5p and alleviated the prohibitory effect of miR-370-3p/miR-136-5p on MYO5A, which accelerated ESCC progression.CONCLUSION:circFNDC3B positively adjusted the MYO5A expression via spongy miR-370-3p/miR-136-5p, hence achieving the cancer-promoting effect on ESCC. circFNDC3B was a prospective diagnosis marker for ESCC.

BACKGROUND:Long-term bed rest in neurointensive care (NIC) patients leads to skeletal muscle atrophy and cognitive dysfunction, which seriously affects the physical fitness and final prognosis of critically ill patients. Exercise therapy plays an increasingly important role in the treatment and rehabilitation of patients with sarcopenia. However, the therapeutic effect and mechanism of exercise therapy for patients with neurological impairment remain unclear.METHODS:Serum samples of NIC patients before and after exercise therapy and normal people were collected to detect interleukin-6 (IL-6) and interleukin-1β levels by enzyme-linked immunosorbent assay (ELISA). Middle cerebral artery occlusion (MCAO) was used for the construction of a rat model. The Morris water maze test, exploration test, and open-field test were used to assess neurological function in rats. Western blot and quantitative real-time polymerase chain reaction were performed to evaluate the activation of IL-6/adenosine-monophosphate-activated protein kinase (AMPK) signaling.RESULTS:Exercise therapy attenuated IL-6 expression in NIC patients. Exercise therapy alleviated cognitive dysfunctions and decreased IL-6 expression in MCAO rats. Exercise therapy alleviated gastrocnemius muscle injury in rats after MCAO by modulating IL-6/AMPK signaling.CONCLUSIONS:Treadmill exercise decreases inflammation in MCAO rats via modulating IL-6/AMPK signaling.

Mechanisms underlying cognitive impairment (CI) in hypertensive patients remain relatively unclear. The present study aimed to explore the relationship among serum exosomal microRNAs (miRNAs), cerebrovascular reactivity (CVR), and cognitive function in hypertensive patients. Seventy-three hypertensive patients with CI (HT-CI), 67 hypertensive patients with normal cognition (HT-NC), and 37 healthy controls underwent identification of exosomal miRNA, multimodal magnetic resonance imaging (MRI) scans, and neuropsychological tests. CVR mapping was investigated based on resting-state functional MRI data. Compared with healthy subjects and HT-NC subjects, HT-CI subjects displayed decreased serum exosomal miRNA-330-3p. The group difference of CVR was mainly found in the left frontal lobe and demonstrated that HT-CI group had a lower CVR than both HT-NC group and control group. Furthermore, both the CVR in the left medial superior frontal gyrus and the miRNA-330-3p level were significantly correlated with executive function ( r = −0.275, P = 0.021, and r = −0.246, P = 0.04, respectively) in HT-CI subjects, and the CVR was significantly correlated with the miRNA-330-3p level ( r = 0.246, P = 0.040). Notably, path analysis showed that the CVR mediated the association between miRNA-330-3p and executive function. In conclusion, decreased miRNA-330-3p might contribute to CI in hypertensive patients by decreasing frontal CVR and could be a biomarker of early diagnosis.

BackgroundLung adenocarcinoma (LUAD) is a leading malignancy and has a poor prognosis over the decades. LUAD is characterized by dysregulation of cell cycle. Immunotherapy has emerged as an ideal option for treating LUAD. Nevertheless, optimal biomarkers to predict outcomes of immunotherapy is still ill-defined and little is known about the interaction of cell cycle-related genes (CCRGs) and immunity-related genes (IRGs).MethodsWe downloaded gene expression and clinical data from TCGA and GEO database. LASSO regression and Cox regression were used to construct a differentially expressed CCRGs and IRGs signature. We used Kaplan-Meier analysis to compare survival of LUAD patients. We constructed a nomogram to predict the survival and calibration curves were used to evaluate the accuracy.ResultsA total of 61 differentially expressed CCRGs and IRGs were screened out. We constructed a new risk model based on 8 genes, including ACVR1B, BIRC5, NR2E1, INSR, TGFA, BMP7, CD28, NUDT6. Subgroup analysis revealed the risk model accurately predicted the overall survival in LUAD patients with different clinical features and was correlated with immune cells infiltration. A nomogram based on the risk model exhibited excellent performance in survival prediction of LUAD.ConclusionsThe 8 gene survival signature and nomogram in our study are effective and have potential clinical application to predict prognosis of LUAD.

Abstract Background Hypoxic tumour microenvironment (TME) is a key regulator in cancer progression. However, the communications between hypoxic cells and other components in TME during colorectal cancer (CRC) progression via extracellular vesicles (EVs) remain unclear. Methods High‐throughput sequencing was employed to detect aberrantly expressed microRNAs (miRNAs) in hypoxic EVs. Quantitative real‐time PCR was used to confirm and screen preliminarily candidate miRNAs. The effects of EVs derived from hypoxia (<1% O2) and miR‐361‐3p on CRC growth were assessed using CCK‐8 assays, colony formation assays, EdU assays, flow cytometric assays and mouse xenograft. Then, the specific mechanisms of miR‐361‐3p were investigated by RNA immunoprecipitation, luciferase reporter assay, Western blot, chromatin immunoprecipitation, immunohistochemistry and rescue experiments. Results The level of miR‐361‐3p expression was remarkably elevated in hypoxic EVs and can be transferred to CRC cells. Functional experiments exhibited that hypoxic EVs facilitated cell growth and suppressed cell apoptosis by transferring miR‐361‐3p of CRC. Hypoxia‐inducible factor‐1α induced the elevation of miR‐361‐3p levels in hypoxic EVs. Upregulated miR‐361‐3p in CRC inhibited cell apoptosis and facilitated cell growth by directly targeting TNF receptor‐associated factor 3, which consequently activated the noncanonical NF‐κB pathway. Moreover, the high expression of circulating exosomal miR‐361‐3p was correlated to worse prognosis of CRC patients. Conclusions Altogether, the abnormality of exosomal miR‐361‐3p derived from hypoxia acts vital roles in the regulation of CRC growth and apoptosis and can be an emerging prognostic biomarker and a therapeutic target for CRC patients.

Background Extracellular vesicles (EVs) are endogenous membrane vesicles with a diameter of 30–200 nm. It has been reported that hypoxic cancer cells can release numerous EVs to mediate multiple regional and systemic effects in the tumor microenvironment. Methods In this study, we used ultracentrifugation to extract EVs secreted by TE‐13, an esophageal squamous carcinoma (ESCC) cell line during normoxia and hypoxia and performed high‐throughput sequencing to detect exosomal miRNAs. Gene ontology (GO) and KEGG pathway analyses were used to reveal pathways potentially regulated by the miRNAs. Results A total of 10 810 miRNAs were detected; 50 were significantly upregulated and 34 were significantly downregulated under hypoxic environment. GO analysis identified enrichment of protein binding, regulation of transcription (DNA‐templated), and membrane as molecular function, biological process, and cellular component, respectively. KEGG pathway analysis revealed cancer‐associated pathways, phospholipase D signaling pathway, autophagy, focal adhesion and AGE‐RAGE signaling as the key pathways. Further verification experiment from qRT‐PCR indicated that miR‐128‐3p, miR‐140‐3p, miR‐340‐5p, miR‐452‐5p, miR‐769‐5p and miR‐1304‐p5 were significantly upregulated in EVs from hypoxia TE‐13 cells while miR‐340‐5p was significantly upregulated in two other ESCC cells, ECA109 and TE‐1. Conclusion This study, for the first time reveals changes in the expression of exosomal miRNAs in hypoxic ESCC cells and these findings will act as a resource to study the hypoxic tumor microenvironment and ESCC EVs.

Introduction: Cerebral small vessel disease (CSVD) refers to a group of clinical syndromes involving cerebral arterioles, perforating arterioles, capillaries and venules, which is characterized by abnormal emotion, abnormal gait, urinary dysfunction, stroke, dementia and so on. It’s five to six times as much as stroke, and accounts for 45% of AD. Early forecast and early intervention can slow down the occurrence and development of cognitive impairment. Ethics number: 2016-134-01; Clinical Registration number: ChiCTR-OOC-17010562. Methods: 195 patients were enrolled in a case-control study and a cohort study. Exosomes were extracted by miRNeasy Serum/Plasma KIT (Qiagen). Transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and exosome reference particle size technique were used to identify exosomes, than, miRNA sequencing was performed and the results were analyzed and verified. Patients underwent neuropsychological measurement including the general cognitive examination and multiple cognitive domain assessments. Results: There were 5 differentially expressed exosome miRNAs between patients with cognitive impairment and the normal subjects, that was miRNA-330/miRNA-339/miRNA-432/miRNA-625/miRNA-6852 were associated with overall cognitive score ( P < 0.001). Among the miRNAs, Significant correlations were observed between miRNA-330 and executive function, as well as processing speed (Pearson’s r = 0.30 after log transformation, P = 0.002; Pearson’s r = 0.24 after log transformation, P = 0.039, respectively). miRNA-432 was significantly correlated to verbal fluency (Pearson’s r = -0.33 after log transformation, P < 0.001). Conclusions: Peripheral blood exosome miRNAs are markers for overall cognitive impairment and correlates with impairment in the domains executive function, verbal fluency and processing speed. It is of great significance for clinical early prevention and early treatment.

Introduction: Microglia are the main type of resident immune cells in the brain. Under different stimuli, microglia exhibit two entirely different functional activated states, termed the pro-inflammatory (M1) phenotype and the anti-inflammatory (M2) phenotype. Almost all cell types can release exosomes, which are considered as important mediators in cell-to-cell communication. Cell-derived exosomal cargo contains a variety of proteins and nucleic acids, which can work locally or stably transferred to recipient cells. Here, we explored the effects of different phenotypes of microglia-derived exosomes on ischemia-induced brain injury. Methods: Primary microglial cells (PMGs) were treated with LPS or IL-4 to induce the M1 or M2 phenotype, respectively. Exosomes were isolated from the cell culture supernatant. Neuronal apoptosis was examined after oxygen-glucose deprivation (OGD) treatment with different exosomes. Transient middle cerebral artery occlusion (MCAO) was used as the in vivo ischemic stroke model to observe the effects of different phenotypes of microglia-derived exosomes on brain injury. Protein profiles were investigated to screen the candidate molecules that mediate distinct functions. Results: Exosomes derived from different phenotypes of microglia have similar diameter, density and exosomal markers. PMGs-derived exosomes can be taken up by neurons. M1 microglia-derived exosomes increased neuronal apoptosis after OGD treatment than OGD alone, while M2 microglia-derived exosomes attenuated neuronal apoptosis. In vivo results revealed that M2 microglia-derived exosomes alleviated brain infarct size and neurological deficits. Besides, M1 microglia-derived exosomes aggravated brain damage. Proteomic analysis showed that the expression of complement-related proteins in M1 microglia-derived exosomes was significantly upregulated than that of M2 microglia-derived exosomes, while the underlying mechanism mediated by specific proteins needs further verification. Conclusions: We demonstrated that M1 microglia-derived exosomes could aggravate ischemic brain injury, while M2 microglia-derived exosomes attenuated this phenomenon. We will explore the detailed mechanisms in the future.

Radiotherapy is one of the primary therapeutic modalities for patients diagnosed esophageal squamous cell carcinoma(ESCC). Previous studies have shown that chemotherapy resistance could be linked with the overexpression vascular ATPases(V-ATPase) subunits genes. However, it is unknown whether V-ATPase subunits genes play a role in radiotherapy resistance. The aim of this study was to investigate the effect of the ATP6V1C1 in radiotherapy resistance. siRNA and plasmids were used to transfect low expression of ATP6V1C1 in TE13 (human ESCC cell) and high expressed in ECA109 (human ESCC cell), respectively. To observe proliferation, radiosensitivity, apoptosis and DNA-damage response, colony formation assays, EDU assays, flow cytometry and γH2AX assay were used with or without radiation exposure, separately. The quantities of the autophagosomes and autolysosomes by immunofluorescence were calculated. Autophagic microstructure were discovered by transmission electron microscopy, and the study also repeated in vivo by nude mice. Western blot assay was applied to prove changes in relative proteins. We found that suppressing ATP6V1C1 increased the sensitivity of ESCC cells after RT. Silencing ATP6V1C1 with IR suppressed the tumor growth and promoted autophagy. Besides, the underlying mechanism of ATP6V1C1, which is not fatally disrupted, is that ATP6V1C1 with ionizing radiation (IR)decreased apoptosis and inhibited autophagy may by activating mTOR signaling to suppress radiosensitivity for ESCC cells. Thus, we first reported that the ATP6V1C1 may represent a potential radiotherapeutic target by effect on radiation sensitivity for ESCC.

BACKGROUND:Based on accumulating evidence, excessive activation of microglia-mediated inflammatory responses plays an essential role in ischemic stroke. Poncirin (Pon) exerts anti-hyperalgesic, anti-osteoporotic and anti-tumor effects on various diseases. However, the roles of Pon in microglial activation and the underlying mechanism have not been elucidated. This study aimed to explore whether Pon inhibits lipopolysaccharide (LPS)-induced microglial neuroinflammation and protects against brain ischemic injury in experimental stroke in mice.METHODS:Primary microglia cells were prepared from the cerebral cortices of 1- to 2-day-old C57BL/6J mice. Murine BV2 cells and primary microglia were stimulated with LPS and the effects of a non-cytotoxic concentration of Pon on LPS-stimulated pro-inflammatory factors were measured using real-time PCR and enzyme-linked immunosorbent assays (ELISAs). Western blot analyses were used for mechanistic studies. In an in vivo study, 8-week-old male C57BL/6J mice were subjected to focal cerebral ischemia through middle cerebral artery occlusion (MCAO). Pon (30 mg/kg, i.p.) or the same volume of saline was administered after the MCAO model was established, and the infarct volume was evaluated using 2,3,5-triphenyltetrazolium chloride (TTC) staining. We also evaluated animal behaviours, the expression of pro-inflammatory cytokines and microglial activation in the ischemic hemisphere.RESULTS:Pon prevented the release of nitric oxide (NO), prostaglandin E2 (PGE2), interleukin (IL)-1β, IL-6 and tumor necrosis factor-alpha (TNF-α) in both BV2 cells and primary microglia stimulated with LPS. The inhibitory effects of Pon were associated with the regulation of the ERK1/2, JNK and nuclear factor kappa B (NF-κB) signaling pathways. In mice that underwent MCAO, Pon administration decreased the lesion size and improved neurological deficits. Furthermore, Pon attenuated the production of inflammatory cytokines mainly by restraining microglial activation after ischemic stroke.CONCLUSIONS:Based on the findings from the present study, Pon provides neuroprotection through its anti-inflammatory effects on microglia and it may be a useful treatment for ischemic stroke.

MicroRNAs (miRNAs) are non-coding small RNA molecules that regulate gene expression at the post-transcriptional/translational level. They act a considerable role not only in the normal progress of development but also in aberrant human diseases, including malignancy. With accumulating proofs of miR-105, the complex role of miR-105 during cancer initiation and progression is gradually emerging. miR-105 acts as a tumor suppressor by inhibiting tumor growth and metastasis or as an oncogene by promoting tumor initiation and invasion, depending on particular tumor contexts and base-pairing genes. In this review, we emphasize the characteristics of miR-105 in cancer to elucidate various deadly tumors and discuss transcriptional regulations that may explain fluctuations in miR-105 expression. This review may provide new ideas for applying miR-105 as a diagnostic and prognostic biomarker.

Salt-inducible kinase (SIK), which belongs to the sucrose non-fermenting 1/AMP-activated protein kinase family, was first discovered in the adrenal cortex of a rat on a high-salt diet. As an isoform of the SIK family, SIK2 modulates various biological functions and acts as a signal transmitter in various pathways. Compared with that in adjacent normal tissues, the expression of SIK2 is significantly higher in multiple types of tumors, which indicates its pivotal effect in oncogenesis. Studies on SIK2 have recently underlined its role in several signaling pathways, including the PI3K-Akt-mTOR pathway, the Hippo-YAP pathway, the LKB1-HDAC axis, and the cAMP-PKA axis. Moreover, a few small-molecule SIK2 inhibitors have been found to be able to rescue the oncogenicity of SIK2 during tumor development and reverse its abnormal activation of downstream pathways. In this mini-review, we discuss the results of in vivo and in vitro studies regarding the SIK2 mechanism in different signaling pathways, particularly their regulation of cancer cells. This work may provide new ideas for targeting SIK2 as a novel therapeutic strategy in tumor therapy.