IGFBP5 Promotes Neuronal Apoptosis in a 6-OHDA-Toxicant Model of Parkinson's Disease by Inhibiting the Sonic Hedgehog Signaling Pathway.

INTRODUCTION: Parkinson's disease (PD) is the most common neurodegenerative disease worldwide. Studies have shown that insulin-like growth factor-binding protein 5 (IGFBP5) may contribute to methamphetamine-induced neurotoxicity and neuronal apoptosis in PC-12 cells and rat striatum. Here, we studied the expression and role of IGFBP5 in the 6-OHDA-toxicant model of PD. METHODS: PC-12 and SH-SY5Y cells were exposed to 50 µm 6-OHDA for 24 h. qRT-PCR, western blotting, CCK-8 assay, EdU staining, annexin V staining, and immunofluorescence were performed to study the effects of IGFBP5-specific siRNAs. The effects of IGFBP5 on a rat 6-OHDA model of PD were confirmed by performing behavioral tests, tyrosine hydroxylase (TH) immunofluorescence staining, and western blotting. RESULTS: In the GSE7621 dataset, IGFBP5 was highly expressed in the substantia nigra tissues of PD patients compared to healthy controls. In PC-12 and SH-SY5Y cells, IGFBP5 was upregulated following 6-OHDA exposure in a dose-dependent manner. Silencing of IGFBP5 promoted PC-12 and SH-SY5Y proliferation and inhibited apoptosis under 6-OHDA stimulation. Silencing of IGFBP5 relieved 6-OHDA-induced TH-positive neuron loss. Hedgehog signaling pathway was predicted as a downstream signaling pathway of IGFBP5. Negative regulation between IGFBP5 and sonic hedgehog (SHH) signaling pathway was confirmed in vitro. The effects of IGFBP5 silencing on SH-SY5Y cells were partially reversed using cyclopamine, a direct inhibitor of the SHH signaling pathway. In addition, silencing of IGFBP5 attenuated motor deficits and neuronal damage in 6-OHDA-induced PD rats. CONCLUSION: Elevated IGFBP5 expression may be involved in 6-OHDA-induced neurotoxicity through regulation of the SHH signaling pathway.

Sinigrin improves cerebral ischaemia-reperfusion injury by inhibiting the TLR4 pathway-mediated oxidative stress.

Cerebral ischaemia-reperfusion (CIR) injury occurs in stroke patients after the restoration of cerebral perfusion. Sinigrin, a phytochemical found in cruciferous vegetables, exhibits strong antioxidant activity. This study investigated the role of sinigrin in oxidative stress using a CIR injury model. The effects of sinigrin were studied in middle cerebral artery occlusion (MCAO) rats and oxygen-glucose deprivation/reoxygenation (OGD/R)-injured SH-SY5Y cells. Sinigrin treatment improved brain injury and neurological deficits induced by MCAO surgery in rats. Sinigrin inhibited apoptosis in brain tissues and SH-SY5Y cells following OGD/R induction. Additionally, sinigrin elevated the levels of superoxide dismutase (SOD), glutathione (GSH) and glutathione peroxidase (GSH-Px) while reducing malondialdehyde (MDA) levels. Furthermore, sinigrin inhibited the toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88) signalling pathway. The anti-apoptotic and antioxidant activities of sinigrin in OGD/R-injured SH-SY5Y cells were reversed by TLR4 overexpression. In conclusion, sinigrin inhibits oxidative stress in CIR injury by suppressing the TLR4/MyD88 signalling pathway.

Hyperthermia is a promising potential adjunct to treating sporotrichosis: A refractory case of HIV sporotrichosis and in vitro experiments on Sporothrix spp.

BACKGROUND: Hyperthermia is a common monotherapy for sporotrichosis, but only in patients with special conditions, such as pregnancy and nursing. However, hyperthermia has not been used more widely for sporotrichosis in clinical practice. PATIENTS/METHODS: An HIV-positive adult male with lymphocutaneous sporotrichosis caused by Sporothrix globosa that did not respond to conventional itraconazole therapy lasting >2 months received adjunctive therapy with local hyperthermia. To simulate the effects of heat exposure on the growth and morphology of Sporothrix spp. in vitro, S. globosa, S. schenckii and S. brasiliensis were exposed to intermittent heat (42°C) for 1 h a day for 7 or 28 days and observed under transmission electron microscopy. RESULTS: Itraconazole combined with local hyperthermia significantly improved the lesions, and the patient was successfully cured of sporotrichosis, with no recurrence after 2 years of follow-up. Cultures of Sporothrix spp. treated with 7 days of daily heat exposure in vitro showed obvious decreases in colony diameters, but not numbers, compared with untreated cultures (p < .001). After 28 days of heat exposure in vitro, Sporothrix spp. were unable to thrive (p < .001), and ultrastructural alterations, including loose cell wall structure, incomplete cell membrane, disrupted vacuoles and fragmented nuclei, were noticeable. CONCLUSIONS: Our case findings and in vitro experiments on Sporothrix spp., together with a literature review of previous sporotrichosis cases, suggest that hyperthermia has a clinical role as a treatment adjunct. Large-scale clinical trials are required to examine the utility of hyperthermia in various forms of cutaneous sporotrichosis.

Sporothrix globosa melanin regulates autophagy via the TLR2 signaling pathway in THP-1 macrophages.

Melanin, an important virulence factor of pathogenic fungi, has been shown to suppress host immune responses in multiple ways. Autophagy is a vital cellular mechanism underlying the host's innate immunity against microbial infections. However, the potential influence of melanin on autophagy has not been explored. We investigated the effect of melanin on autophagy in macrophages, which play a key role in controlling Sporothrix spp. infection, as well as the mechanism of melanin interaction with Toll-like receptor (TLR)-induced pathways. Sporothrix globosa conidia (wild-type and melanin-deficient mutant strains) or yeast cells were co-cultured with THP-1 macrophages to demonstrate that, although S. globosa infection led to the activation of autophagy-related proteins and increased autophagic flux, S. globosa melanin suppressed macrophage autophagy. Incubation with S. globosa conidia also increased the expression levels of reactive oxygen species and multiple proinflammatory cytokines (interleukin-6, tumor necrosis factor-α, interleukin-1ß and interferon-γ) in macrophages. These effects were attenuated as melanin presented. Furthermore, while S. globosa conidia significantly increased the expression of both TLR2 and TLR4 in macrophages, the knockdown of TLR2, but not TLR4, with small interfering RNA suppressed autophagy. Overall, this study revealed the novel immune defense ability of S. globosa melanin to inhibit macrophage functionality by resisting macrophage autophagy through the regulation of TLR2 expression.

ZBP1 mediates the progression of Alzheimer's disease via pyroptosis by regulating IRF3.

Alzheimer's disease (AD) is one of the leading causes of death throughout the world. Z-DNA binding protein 1 (ZBP1), a DNA-related gene, is associated with inflammation, and its expression is altered in AD brain. We aimed to elucidate the exact role of ZBP1 in AD development and its potential regulatory mechanism. First, we constructed both in vivo and in vitro models of AD and investigated the ZBP1 expression profile. A loss-of-function assay was performed by transfecting lentivirus carrying ZBP1 short hairpin RNA (shRNA). By evaluating cell death, oxidative stress, inflammation response and pyroptosis, the function of ZBP1 was validated. Finally, the correlation between ZBP1 and interferon regulatory factor 3 (IRF3) was verified. We also performed rescue experiments to validate the crucial role of IRF3 in ZBP1-mediated AD progression. According to our results, ZBP1 was upregulated in AD rat tissue and AD neurons. Silencing ZBP1 dramatically decreased cell injury, oxidative stress and inflammation in AD neurons and improved the cognitive function of AD rats. Additionally, IRF3 expression and phosphorylation were significantly elevated during AD development and positively correlated with ZBP1. Taken together, silencing ZBP1 suppressed cell injury and pyroptosis of AD neurons and improved cognitive function of AD rats via inhibiting IRF3. These findings might provide a novel insight for AD target diagnosis and therapy.

AcneTyper: An automatic diagnosis method of dermoscopic acne image via self-ensemble and stacking.

BACKGROUND: Acne is a skin lesion type widely existing in adolescents, and poses computational challenges for automatic diagnosis. Computer vision algorithms are utilized to detect and determine different subtypes of acne. Most of the existing acne detection algorithms are based on the facial natural images, which carry noisy factors like illuminations. OBJECTIVE: In order to tackle this issue, this study collected a dataset ACNEDer of dermoscopic acne images with annotations. Deep learning methods have demonstrated powerful capabilities in automatic acne diagnosis, and they usually release the training epoch with the best performance as the delivered model. METHODS: This study proposes a novel self-ensemble and stacking-based framework AcneTyper for diagnosing the acne subtypes. Instead of delivering the best epoch, AcneTyper consolidates the prediction results of all training epochs as the latent features and stacks the best subset of these latent features for distinguishing different acne subtypes. RESULTS: The proposed AcneTyper framework achieves a promising detection performance of acne subtypes and even outperforms a clinical dermatologist with two-year experiences by 6.8% in accuracy. CONCLUSION: The method we proposed is used to determine different subtypes of acne and outperforms inexperienced dermatologists and contributes to reducing the probability of misdiagnosis.

Edaravone Attenuates Aß 1-42-Induced Inflammatory Damage and Ferroptosis in HT22 Cells.

Ferroptosis and neuroinflammation play a crucial role in the pathogenesis of Alzheimer's disease (AD), and Edaravone (EDA) has been demonstrated to have anti-inflammatory, antioxidant and neuroprotective effects in neurodegenerative diseases. However, the relationship between EDA and ferroptosis in AD is unidentified. This research aimed to elucidate the mechanism of EDA in AD with Aß 1-42-induced HT22 cells as in vitro cell model. The results showed that EDA could significantly reduce Aß1-42-induced apoptosis of HT22 cells and formation of pro-inflammatory factors TNF-α, IL-1ß and IL-6, prevent the activation of TLR4/NF-κB /NLRP3 signaling pathway, and inhibit ferroptosis and lipid peroxidation. Taken together, EDA contributes to inhibiting neuroinflammatory injury and ferroptosis in Aß 1-42-induced HT22 cells, and thus may be a potential candidate for the treatment of AD.

Effectiveness comparisons of various therapies for FIGO stage IB2/IIA2 cervical cancer: a Bayesian network meta-analysis.

BACKGROUND: Cervical cancer is a common malignancy of the female genital tract. Treatment options for cervical cancer patients diagnosed at FIGO (2009) stage IB2 and IIA2 remains controversial. METHODS: We perform a Bayesian network meta-analysis to directly or indirectly compare various interventions for FIGO (2009) IB2 and IIA2 disease, in order to improve our understand of the optimal treatment strategy for these women. Three databases were searched for articles published between 1971 and 2020. Data on included study characteristics, outcomes, and risk of bias were abstracted by two reviewers. RESULTS: Seven thousand four hundred eighty-six articles were identified. Thirteen randomized controlled trials of FIGO (2009) IB2 and IIA2 cervical cancer patients were included in the final analysis. These trials used six different interventions: concomitant chemoradiotherapy (CCRT), radical surgery (RS), radical surgery following chemoradiotherapy (CCRT+RS), neoadjuvant chemotherapy followed by radical surgery (NACT+RS), adjuvant radiotherapy followed by Radical surgery (RT + RS), radiotherapy alone (RT).SUCRA ranking of OS and Relapse identified CCRT+RS and CCRT as the best interventions, respectively. Systematic clustering analysis identified the CCRT group as a unique cluster. CONCLUSION: These data suggest that CCRT may be the best approach for improving the clinical outcome of cervical cancer patients diagnosed at FIGO (2009) stage IB2/IIA2. Phase III randomized trials should be performed in order to robustly assess the relative efficacy of available treatment strategies in this disease context.

In vitro antifungal susceptibility of Sporothrix globosa isolates from Jilin Province, northeastern China: comparison of yeast and mycelial phases.

The dimorphic fungus Sporothrix globosa is the predominant etiologic agent causing sporotrichosis in China, particularly in the northeast. It has been demonstrated that the incubation temperature and growth phase can influence in vitro antifungal susceptibility profiles of S. schenckii sensu stricto and S. brasiliensis (sibling species of S. globosa). Few studies have reported on the antifungal susceptibility of S. globosa, especially using large numbers of isolates. In this study, we assessed the susceptibility of 80 isolates of S. globosa originating from Jilin Province, northeastern China, to six antifungal agents (itraconazole, terbinafine, voriconazole, posaconazole, fluconazole, and amphotericin B), at varying incubation temperatures and in different fungal growth phases. The isolates were most sensitive to terbinafine (geometric mean [GM] of the minimum inhibitory concentration [MIC]: 0.0356 µg/ml for the mycelial phase at 30 °C, 0.0332 µg/ml for the mycelial phase at 35 °C, and 0.031 µg/ml for the yeast phase, respectively), followed by posaconazole (GM of the MIC: 4.2501 µg/ml for the mycelial phase at 30 °C, 1.4142 µg/ml for the mycelial phase at 35 °C, and 0.7195 µg/ml for the yeast phase, respectively) and itraconazole (GM of the MIC: 6.8448 µg/ml for the mycelial phase at 30 °C, 3.1383 µg/ml for the mycelial phase at 35 °C, and 1.0263 µg/ml for the yeast phase, respectively). The isolates were relatively resistant to fluconazole (GM of the MIC: 76.7716 µg/ml for the mycelial phase at 30 °C, 66.2570 µg/ml for the mycelial phase at 35 °C, and 24.4625 µg/ml for the yeast phase, respectively) and voriconazole (GM of the MIC: 26.2183 µg/ml for the mycelial phase at 30 °C, 13.6895 µg/ml for the mycelial phase at 35 °C, and 1.3899 µg/ml for the yeast phase, respectively). For all the tested azole drugs, the MICs at 30 °C were significantly higher than those at 35 °C (P < 0.001); for all agents except terbinafine, the MICs of S. globosa in the yeast phase were significantly lower than those of the strains in the mycelial phase (P < 0.001). These results show that the sensitivities of S. globosa to antifungal compounds are dependent on incubation temperature and growth phase. To the best of our knowledge, this is the largest study of antifungal susceptibility of S. globosa isolates reported to date. To establish epidemiological cutoff values for S. globosa, further antifungal susceptibility testing studies by independent laboratories located in different regions and using uniform conditions are required.

Aggressive sebaceous carcinoma of the scalp: a case report and literature review.

Sebaceous carcinoma (SC) is an uncommon and potentially aggressive adnexal neoplasm. SC presents most often as a periocular tumor arising from the ocular adnexa with unclear pathogenesis. Aggressive SC of the scalp is extremely rare. Here, we describe a rare case of aggressive SC of the scalp in a 61-year-old female, who presented with a scalp neoplasm lasting for two months. Cranial magnetic resonance imaging (MRI) revealed a partly necrotic and cystic tumor with mixed signal shadow. The parietal multiple destructive lesions invaded the skull, involved the dura mater, and compressed the associated superior sagittal sinus. A wide local lesion excision with I stage repair of dura mater and I stage bone graft of skull was performed in our department. The patient was treated with external radiation to prevent recurrence and was followed for 3 years with favorable results. The relevant literature regarding aggressive SC was reviewed, and the clinical manifestations, radiological characteristics, surgical strategies, histopathological findings, and prognosis are discussed. This cutaneous malignancy invaded skull, dura mater or brain, or metastasized to the lymph nodes and viscera, with high recurrence and fatality rates. It is necessary to be aware of these rare examples which showed unexpectedly clinical behaviors. Early precise diagnosis and improved radical treatment remain essential steps against aggressive cutaneous malignancy. Patients with aggressive SC of the scalp should be closely followed to assess recurrence and distant metastasis.

Th2 Biased Immunity With Altered B Cell Profiles in Circulation of Patients With Sporotrichosis Caused by Sporothrix globosa.

Sporotrichosis is a subcutaneous mycotic infection, and Sporothrixglobosa is one of the causative agents with a worldwide distribution, notably in Asia. However, the immune profile in human sporotrichosis caused by S. globosa still remains obscure. Here, we demonstrated enhanced Th2 response in circulation with significant increases in Th2 frequency, Th2/Tregs as well as IL-4 seretion in patients. Elevated IL-17A+Th17 percentage was accompanied with reduced IL-17A level in serum, which may imply a dysfunction of this CD4+T subset in S. globosa infection. In addition, Th2 percentage, the ratios of Th2/Tregs and Th17/Tregs were all raised in patients with fixed cutaneous form, while only Th2/Tregs displayed increment in lymphocutaneous form. Meanwhile, the percentage of double negative B cells was significantly increased and positively correlated with Th2 and Tregs in whole patients. Except naïve B cells, all memory B cells together with Th2 cells increased in patients with short duration (less than 6 months), which may suggest a collaboration of T cells with altered B cell profile in human sporotrichosis caused by S. globosa. In consistent with the changes of IFN-γ+Th1, IL-4+Th2 and IL-17A+Th17 in patients with short duration, the percentages of these effector T cells all expanded when cocultured with S. globosa yeast cells in vitro. These data shed light on the potential involvement of peripheral T and B cell immunity against this mycotic infection and indicated that different immune responses existed in different stages of sporotrichosis; meanwhile different immune profile may contribute to different clinical manifestations of this disease.

5-Aminolevulinic acid-mediated photodynamic therapy has effective antifungal activity against Sporothrix globosa in vitro.

BACKGROUND: An alternative therapy for sporotrichosis is necessary to reduce the treatment time and raise clinical efficacy. The 5-Aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) is a promising platform with which to treat mycoses. However, despite the worldwide prevalence of Sporothrix globosa, a causative agent of sporotrichosis, the effect of ALA-PDT on this pathogen has not been validated. OBJECTIVES: The aim of this study was to evaluate the effect of ALA-PDT on S globosa and the protection of melanin through an in vitro study. The mechanisms involved were also investigated. METHODS: To estimate the survival rate of S globosa treated with ALA-PDT and the protection offered by melanin, the conidia and yeast cells of wild-type S globosa (Mel+), other clinical strains, tricyclazole-treated Mel+ and an albino mutant strain (Mel-) were incubated with and without ALA or irradiation. Reactive oxygen species generation by Mel+ conidia induced by ALA-PDT was assayed. SEM and TEM were conducted to obverse ultrastructural changes in the conidia. A comet assay was performed to evaluate DNA damage. RESULTS: The survival rate of S globosa conidia and yeast cells significantly decreased following incubation with 1.19M ALA and 162 J/cm2 irradiation in vitro. Melanin was not only capable of protecting the conidia against ALA-PDT, but also against ALA or irradiation alone. After induction by ALA-PDT, alterations in reactive oxygen species generation, DNA damage and ultrastructural changes were observed. CONCLUSIONS: ALA-PDT inhibits the survival of S globosa conidia in vitro and therefore has potential for the treatment of sporotrichosis.

Comorbid SUNCT Syndrome and Opalski Syndrome Caused by Dorsolateral Medullary Infarction.

Opalski syndrome is a rare variation of lateral medullary syndrome (LMS) accompanied by ipsilateral hemiparesis. Short-lasting unilateral neuralgiform headaches with conjunctival injection and tearing (SUNCT) is a rare headache syndrome which belongs to the trigeminal autonomic cephalalgias. SUNCT syndrome has been previously described in association with LMS. We here describe a case of SUNCT syndrome with Opalski syndrome caused by dorsolateral medullary infarction.

Che-1 inhibits oxygen-glucose deprivation/reoxygenation-induced neuronal apoptosis associated with inhibition of the p53-mediated proapoptotic signaling pathway.

Accumulating evidence suggests that Che-1 is a strong antiapoptotic protein and can protect cells against various insults. However, whether Che-1 is involved in the protection of neurons against cerebral ischemia/reperfusion injury remains unclear. In this study, we aimed to investigate the potential role of Che-1 in regulating cerebral ischemia/reperfusion injury-induced neuronal injury using the oxygen-glucose deprivation and reoxygenation (OGD/R) model in vitro. We found that Che-1 expression was induced in neurons following OGD/R treatment. Functional experiments showed that Che-1 knockdown aggravated OGD/R-induced neuronal apoptosis. In contrast, Che-1 overexpression exerted a protective effect against OGD/R-induced neuronal apoptosis. Moreover, our results showed that the protective effect of Che-1 was associated with the inhibition of p53-mediated proapoptotic genes, including Puma, Noxa, and Bax. In addition, we showed that Che-1 impeded the transcript activity of p53 toward apoptosis. Taken together, our results indicate that Che-1 alleviates OGD/R-induced neuronal apoptosis in-vitro through inhibition of p53-mediated proapoptotic signaling. Our study suggests that Che-1 may serve as a potential therapeutic target for the treatment of cerebral ischemic/reperfusion injury in vivo.

Genetic variation of Sporothrix globosa isolates from diverse geographic and clinical origins in China.

Sporothrix globosa is the main causative agent of sporotrichosis, a common mycosis that usually affects the skin, in China. Despite increasing efforts in the molecular identification of this fungal pathogen, its modes of transmission and epidemiology remain poorly understood. The goals of this study were to assess the genetic diversity of S. globosa using amplified fragment length polymorphism (AFLP) analysis and to assess the correlation of AFLP profiles with the geographic origins, growth rates, clinical forms, and antifungal susceptibilities of S. globosa isolates. AFLP analysis of 225 clinical S. globosa isolates from eight provinces or municipalities in China identified eight distinct clustering groups (I-VIII), with groups I, II and IV being the most common. The AFLP genotypes showed distinct distribution patterns among different regions within Jilin Province and between northern and southern China, but there was no obvious association between the AFLP genotypes and the growth rates, clinical forms or antifungal susceptibilities of the S. globosa isolates. These results expand our understanding of the genetic variation of S. globosa and suggest that AFLP analysis is a potentially useful tool for studying the epidemiology of this fungal pathogen.