Validation of different predictive scoring scales in patients with new-onset epileptic seizures or epilepsy related to neuronal surface antibody-mediated autoimmune encephalitis.

OBJECTIVE: To validate the different predictive scoring scales in the Chinese population with new-onset epileptic seizures or epilepsy of unknown etiology related to neuronal surface antibody (Ab)-mediated autoimmune encephalitis (AE). METHODS: We retrospectively reviewed the charts of 174 consecutive patients from October 2018 to December 2022, whose serum and cerebrospinal fluid samples were tested for neuronal surface Abs. The antibody prevalence in epilepsy and encephalopathy (APE2), antibodies contributing to focal epilepsy signs and symptoms (ACES), "obvious" indications for neural antibody testing in epilepsy or seizures (ONES) checklist, and the combinations were used to validate the predictive models of neuronal surface Ab-mediated AE. RESULTS: A total of 139 patients with new-onset epileptic seizures or epilepsy of unknown etiology were enrolled. Abs were detected in 37 patients (26.6%). The APE2/ONES reflex score had the highest sensitivity (89.2%) and lowest specificity (41.7%). The ACES score had the lowest sensitivity (67.5%) and highest specificity (64.7%). Variations in the performance were observed in the different types of AE. 100% of patients with anti-γ-aminobutyric acid B-B receptor encephalitis were predicted by ONES, APE2/ONES reflex, and ACES/ONES reflex scores. Only 75% of patients with anti-N-methyl-D-aspartate receptor encephalitis were predicted by the APE2/ONES and ACES/ONES reflex scores. CONCLUSION: Our study was the first to validate various predictive scoring scales in the Chinese cohort of patients with new-onset epileptic seizures or epilepsy of unknown etiology related to neuronal surface Ab-mediated AE. Based upon clinical suspicion, more than one scoring scale should be performed to predict the chance of AE in those patients.

Active fluorescent modulation for low-noise super-resolution microscopy.

Extracting the position of individual molecular probes with high precision is the basis and core of super-resolution microscopy. However, with the expectation of low-light conditions in life science research, the signal-to-noise ratio (SNR) decreases and signal extraction faces a great challenge. Here, based on temporally modulating the fluorescence emission at certain periodical patterns, we achieved super-resolution imaging with high sensitivity by largely suppressing the background noise. We propose simple bright-dim (BD) fluorescent modulation and delicate control by phase-modulated excitation. We demonstrate that the strategy can effectively enhance signal extraction in both sparsely and densely labeled biological samples, and thus improve the efficiency and precision of super-resolution imaging. This active modulation technique is generally applicable to various fluorescent labels, super-resolution techniques, and advanced algorithms, allowing a wide range of bioimaging applications.

Speech2EEG: Leveraging Pretrained Speech Model for EEG Signal Recognition.

Identifying meaningful brain activities is critical in brain-computer interface (BCI) applications. Recently, an increasing number of neural network approaches have been proposed to recognize EEG signals. However, these approaches depend heavily on using complex network structures to improve the performance of EEG recognition and suffer from the deficit of training data. Inspired by the waveform characteristics and processing methods shared between EEG and speech signals, we propose Speech2EEG, a novel EEG recognition method that leverages pretrained speech features to improve the accuracy of EEG recognition. Specifically, a pretrained speech processing model is adapted to the EEG domain to extract multichannel temporal embeddings. Then, several aggregation methods, including the weighted average, channelwise aggregation, and channel-and-depthwise aggregation, are implemented to exploit and integrate the multichannel temporal embeddings. Finally, a classification network is used to predict EEG categories based on the integrated features. Our work is the first to explore the use of pretrained speech models for EEG signal analysis as well as the effective ways to integrate the multichannel temporal embeddings from the EEG signal. Extensive experimental results suggest that the proposed Speech2EEG method achieves state-of-the-art performance on two challenging motor imagery (MI) datasets, the BCI IV-2a and BCI IV-2b datasets, with accuracies of 89.5% and 84.07% , respectively. Visualization analysis of the multichannel temporal embeddings show that the Speech2EEG architecture can capture useful patterns related to MI categories, which can provide a novel solution for subsequent research under the constraints of a limited dataset scale.

Enhancing Weak-Signal Extraction for Single-Molecule Localization Microscopy.

Single-molecule localization microscopy (SMLM) has been widely used in biological imaging due to its ultrahigh spatial resolution. However, due to the strategy of reducing photodamage to living cells, the fluorescence signals of emitters are usually weak and the detector noises become non-negligible, which leads to localization misalignments and signal losses, thus deteriorating the imaging capability of SMLM. Here, we propose an active modulation method to control the fluorescence of the probe emitters. It actually marks the emitters with artificial blinking character, which directly distinguishes weak signals from multiple detector noises. We demonstrated from simulations and experiments that this method improves the signal-to-noise ratio by about 10 dB over the non-modulated method and boosts the sensitivity of single-molecule localization down to -4 dB, which significantly reduces localization misalignments and signal losses in SMLM. This signal-noise decoupling strategy is generally applicable to the super-resolution system with versatile labeled probes to improve their imaging capability. We also showed its application to the densely labeled sample, showing its flexibility in super-resolution nanoscopy.

SIN: Semantic Inference Network for Few-Shot Streaming Label Learning.

Streaming label learning aims to model newly emerged labels for multilabel classification systems, which requires plenty of new label data for training. However, in changing environments, only a small amount of new label data can practically be collected. In this work, we formulate and study few-shot streaming label learning (FSLL), which models emerging new labels with only a few annotated examples by utilizing the knowledge learned from past labels. We propose a meta-learning framework, semantic inference network (SIN), which can learn and infer the semantic correlation between new labels and past labels to adapt FSLL tasks from a few examples effectively. SIN leverages label semantic representation to regularize the output space and acquires labelwise meta-knowledge based on gradient-based meta-learning. Moreover, SIN incorporates a novel label decision module with a meta-threshold loss to find the optimal confidence thresholds for each new label. Theoretically, we illustrate that the proposed semantic inference mechanism could constrain the complexity of hypotheses space to reduce the risk of overfitting and achieve better generalizability. Experimentally, extensive empirical results and ablation studies demonstrate the performance of SIN is superior to the prior state-of-the-art methods on FSLL.

Resolving plasmonic hotspots by label-free super-resolution microscopy.

The plasmonic hotspot of metal nanostructures has small dimension far beyond the optical diffraction limit. When trying to locate the hotspot using fluorescent probes, the localization is significantly distorted due to the coupling of emission and surface plasmon. A label-free technique can solve the problem, which uses hotspot emission as the native probe. We demonstrate a super-resolution microscopy investigation based on this idea. By modulating hotspot emission of crossed silver nanowires, which have a pair of plasmonic hotspots approximately 100 nm apart at the intersection, we precisely locate and separate them with nanometer precision. This label-free technique could be applied for analyzing hotspot distribution with high efficiency and precision.

Transformation of Fonsecaea pedrosoi into sclerotic cells links to the refractoriness of experimental chromoblastomycosis in BALB/c mice via a mechanism involving a chitin-induced impairment of IFN-γ production.

Fonsecaea pedrosoi (F. pedrosoi) is the most common agent of chromoblastomycosis. Transformation of this fungus from its saprophytic phase into pathogenic sclerotic cells in tissue is an essential link to the refractoriness of this infection. Experimental studies in murine models have shown that the absence of CD4+ T cells impairs host defense against F. pedrosoi infection. Clinical research has also suggested that a relatively low level of the Th1 cytokine INF-γ and inefficient T cell proliferation are simultaneously present in patients with severe chromoblastomycosis upon in vitro stimulation with ChromoAg, an antigen prepared from F. pedrosoi. In the present study, we show that in mice intraperitoneally infected with F. pedrosoi-spores, -hyphae or in vitro-induced sclerotic cells respectively, the transformation of this causative agent into sclerotic cells contributes to a compromised Th1 cytokine production in the earlier stage of infection with impaired generation of neutrophil reactive oxygen species (ROS) and pan-inhibition of Th1/Th2/Th17 cytokine production with disseminated infection in the later stage by using a CBA murine Th1/Th2/Th17 cytokine kit. In addition, we have further demonstrated that intraperitoneal administration of recombinant mouse IFN-γ (rmIFN-γ) effectively reduces the fungal load in the infected mouse spleen, and dampens the peritoneal dissemination of F. pedrosoi-sclerotic cells. Meanwhile, exogeneous rmIFN-γ contributes to the formation and maintenance of micro-abscess and restores the decrease in neutrophil ROS generation in the mouse spleen infected with F. pedrosoi-sclerotic cells. Of note, we have once again demonstrated that it is a chitin-like component, but not ß-glucans or mannose moiety, that exclusively accumulates on the outer cell wall of F. pedrosoi-sclerotic cells which were induced in vitro or isolated from the spleens of intraperitoneally infected BALB/c mice. In addition, our results indicate that decreased accumulation of chitin on the surface of live F. pedrosoi-sclerotic cells after chitinase treatment can be self-compensated in a time-dependent manner. Importantly, we have for the first time demonstrated that exclusive accumulation of chitin on the transformed sclerotic cells of F. pedrosoi is involved in an impaired murine Th1 cytokine profile, therefore promoting the refractoriness of experimental murine chromoblastomycosis.

Cerebriform connective tissue nevus of lumbar.

Connective tissue nevi represents a kind of hamartoma, and coalescence of the lesions in a cerebriform mode in the lumbar region without Proteus syndrome is rarely seen. Here, we report a 26-year-old woman presenting with nodules and plaques in her left lumbar region of 26 years in duration. Histopathological examination and Masson-trichrome stain showed increased dermal collagen bundles in a haphazard array. The diagnosis of connective tissue nevi was made. This is the first case report on cerebriform connective tissue nevi without Proteus syndrome in the lumbar region.

A chitin-like component on sclerotic cells of Fonsecaea pedrosoi inhibits Dectin-1-mediated murine Th17 development by masking ß-glucans.

Fonsecaea pedrosoi (F. pedrosoi), a major agent of chromoblastomycosis, has been shown to be recognized primarily by C-type lectin receptors (CLRs) in a murine model of chromoblastomycosis. Specifically, the ß-glucan receptor, Dectin-1, mediates Th17 development and consequent recruitment of neutrophils, and is evidenced to have the capacity to bind to saprophytic hyphae of F. pedrosoi in vitro. However, when embedded in tissue, most etiological agents of chromoblastomycosis including F. pedrosoi will transform into the sclerotic cells, which are linked to the greatest survival of melanized fungi in tissue. In this study, using immunocompetent and athymic (nu/nu) murine models infected subcutaneously or intraperitoneally with F. pedrosoi, we demonstrated that T lymphocytes play an active role in the resolution of localized footpad infection, and there existed a significantly decreased expression of Th17-defining transcription factor Rorγt and inefficient recruitment of neutrophils in chronically infected spleen where the inoculated mycelium of F. pedrosoi transformed into the sclerotic cells. We also found that Dectin-1-expressing histocytes and neutrophils participated in the enclosure of transformed sclerotic cells in the infectious foci. Furthermore, we induced the formation of sclerotic cells in vitro, and evidenced a significantly decreased binding capacity of human or murine-derived Dectin-1 to the induced sclerotic cells in comparison with the saprophytic mycelial forms. Our analysis of ß-glucans-masking components revealed that it is a chitin-like component, but not the mannose moiety on the sclerotic cells, that interferes with the binding of ß-glucans by human or murine Dectin-1. Notably, we demonstrated that although Dectin-1 contributed to the development of IL-17A-producing CD3+CD4+ murine splenocytes upon in vitro-stimulation by saprophytic F. pedrosoi, the masking effect of chitin components partly inhibited Dectin-1-mediated Th17 development upon in vitro-stimulation by induced sclerotic cells. Therefore, these findings extend our understanding of the chronicity of chromoblastomycosis.

Analysis of Th1/Th2 response pattern for erythrodermic psoriasis.

As one of the most serious types of psoriasis, pathogenesis of erythrodermic psoriasis (EP) is unclear so far. In this study, we aimed to detect the levels of Th1/Th2 cytokine-associated transcription factors and T-lymphocyte clone in peripheral blood mononuclear cells (PBMCs) derived from EP patients, and gene expression level of T-bet/GATA-3 in skin lesion. The potential role of Th1/Th2 reaction pattern played in the pathogenesis of EP was also discussed. Serum levels of IFN-γ, IL-2, IL-4 and IL-10 were quantified by ELISA among 16 EP patients, 20 psoriasis vulgaris (PV) patients and 15 healthy controls. The expression levels of T-bet/GATA-3 in the skin lesion and PBMCs were examined by real-time qPCR. The ratio of Th1/Th2 was measured by flow cytometry. The levels of IFN-γ, IL-2, IL-4 and IL-10 were higher in EP patients than in the healthy controls. The levels of IL-4 and IL-10 were 69.44±11.45 and 12.62±4.57 pg/mL, respectively, in EP patients, significantly higher than those in PV patients and healthy controls (P<0.05). Flow cytometry revealed the levels of both Th1 and Th2 in PBMCs from EP patients were higher than those in healthy controls, and the Th1/Th2 ratio was dramatically lower than in PV patients (P<0.01). The ratios of IFN-γ/IL-4 and T-bet/GATA-3 in EP patients were both less than 1.0, suggesting a reversal when compared with the other two groups. Our study indicated that the EP patients exerted a Th1/Th2 bidirectional response pattern, and the balance of Th cell subsets inclines to Th2, which might be one of the important mechanisms of EP pathogenesis.

Progesterone suppresses Th17 cell responses, and enhances the development of regulatory T cells, through thymic stromal lymphopoietin-dependent mechanisms in experimental gonococcal genital tract infection.

In most female patients, the symptoms of genital infection due to Neisseria gonorrhoeae tend to be slight or even absent. Our previous studies suggested that progesterone might play a role in female asymptomatic gonococcal infection. In this study, we demonstrated that progesterone induced the expression of thymic stromal lymphopoietin (TSLP) and regulatory T cells (Treg)-related transcription factor Foxp3, and inhibited the expression of Th17 related transcription factor RORγt, and reduced the influx of neutrophils in murine vaginal gonococcal infection. Blockade of TSLP with antibody partially reversed the effects of progesterone on the murine model of gonococcal vaginal infection. In in vitro experiments, progesterone induced a rapid up-regulation of TSLP in vaginal epithelial cells stimulated with N. gonorrhoeae. Blocking thymic stromal lymphopoietin receptor (TSLPR) with a TSLPR monoclonal antibody partially prevented progesterone suppression of IL-17-producing T cells differentiation, and progesterone promotion of CD4⁺CD25⁺Foxp3⁺ regulatory T cells differentiation. Altogether, our results indicate that the progesterone suppresses Th17 cell responses, and enhances the development of Treg cells, through TSLP-dependent mechanisms, and play a role in female asymptomatic gonococcal infections.

Generalized subcutaneous phaeohyphomycosis caused by Phialophora verrucosa: report of a case and review of literature.

We report a case of subcutaneous phaeohyphomycosis due to Phialophora verrucosa in a 64-year-old Chinese farmer suffering from CD4+ lymphopenia. He presented with diffuse and infiltrated plaques involving the entire face including the eyes, neck, occiput, and extending to the dorsal regions of his torso. The patient is notable for the discrete multifocal nature of the illness in the absence of disseminated infection and rarity of P. verrucosa as a cause of subcutaneous phaeohyphomycosis.

MBL-mediated opsonophagocytosis of Candida albicans by human neutrophils is coupled with intracellular Dectin-1-triggered ROS production.

Mannan-binding lectin (MBL), a lectin homologous to C1q, greatly facilitates C3/C4-mediated opsonophagocytosis of Candida albicans (C. albicans) by human neutrophils, and has the capacity to bind to CR1 (CD35) expressed on circulating neutrophils. The intracellular pool of neutrophil Dectin-1 plays a critical role in stimulating the reactive oxygen species (ROS) generation through recognition of ß-1,3-glucan component of phagocytized zymosan or yeasts. However, little is known about whether MBL can mediate the opsonophagocytosis of Candida albicans by neutrophils independent of complement activation, and whether MBL-mediated opsonophagocytosis influence the intracellular expression of Dectin-1 and ROS production. Here we showed that the inhibited phagocytic efficiency of neutrophils as a result of blockage of Dectin-1 was compensated by exogenous MBL alone in a dose-dependent manner. Furthermore, the expressions of Dectin-1 at mRNA and intracellular protein levels were significantly up-regulated in neutrophils stimulated by MBL-pre-incubated C. albicans, while the expression of surface Dectin-1 remained almost unchanged. Nevertheless, the stimulated ROS production in neutrophils was partly and irreversibly inhibited by blockage of Dectin-1 in the presence of exogenous MBL. Confocal microscopy examination showed that intracellular Dectin-1 was recruited and co-distributed with ROS on the surface of some phagocytized yeasts. The ß-1,3-glucanase digestion test further suggested that the specific recognition and binding site of human Dectin-1 is just the ß-1,3-glucan moiety on the cell wall of C. albicans. These data demonstrate that MBL has an ability to mediate the opsonophagocytosis of Candida albicans by human neutrophils independent of complement activation, which is coupled with intracellular Dectin-1-triggered ROS production.

A combined assay of hTERT and E6 oncoprotein to identify virus-infected keratinocytes with higher telomerase activity in human papillomaviruses 16 and 18-related bowenoid papulosis.

In the present study, we aim to evaluate the application potential of a combined assay of human telomerase reverse transcriptase (hTERT) and E6 oncoprotein in screening the virus-infected keratinocytes with higher telomerase activity in human papillomaviruses (HPV) 16- and 18-related bowenoid papulosis (BP). HPV16/18 DNA in BP (n = 123) was identified by in situ hybridization, the expression of hTERT and E6 in HPV16/18-related BP (n = 68) was determined by immunohistochemistry. We demonstrated that the expression of hTERT correlated well with that of E6 oncoprotein in HPV16/18-related BP lesions (Spearman rho = 0.868, P < 0.01). Furthermore, the majority of keratinocytes with positive nuclear staining for hTERT or E6 in the consecutive sections of each HPV16/18-related BP lesion showed nuclear paleomorphism or nuclear mitosis. In conclusion, we suggested that a combined assay of hTERT and E6 oncoprotein can be used to screen the HPV-infected keratinocytes with higher telomerase activity in HPV16-related and HPV18-related BP lesions.

Genotyping of human adenoviruses using a PCR-based reverse line blot hybridisation assay.

OBJECTIVES: Human adenoviruses are common pathogens associated with a broad spectrum of disease. There is a growing clinical interest in typing clinical isolates since it is becoming increasingly clear that individual serotypes are associated with different disease spectra, virulence, severity of consequences, and outbreaks. Current methods cannot detect all known adenoviruses simultaneously and rapidly. We designed a practical adenovirus typing method with polymerase chain reaction (PCR)-based reverse line blot hybridisation assay (RLB) using hypervariable region-7 (HVR-7) in the hexon gene. METHODS: A PCR-RLB assay was developed based on HVR-7 in the hexon region for potentially genotyping 51 adenovirus serotypes by hybridisation of 62 genotype-specific probes using amplicons generated from one genus-specific primer pair. Single PCR and sequencing were performed for confirmation of RLB results. Eighty-seven previously serotyped clinical isolates (representing 28 serotypes) were studied. RESULTS: Thirty-two different genotypes were detected by RLB from 87 adenovirus isolates, of which 82 isolates showed consistent results with sequencing. Another five isolates revealed evidence by RLB of co-infection, and were confirmed with a combination of genotype-specific single PCR and sequencing. CONCLUSIONS: In comparison to sequencing and serological methods, the advantages of the RLB assay include: (1) rapid genotyping of multiple samples in a single run; (2) successful detection of co-infection; (3) detection of subgenotype variants. This will allow rapid and inexpensive characterisation of adenovirus infections and outbreaks.

[Observation of therapeutic effect on perineal, crissal and progenital pruritus treated with acupuncture according to differentiation].

OBJECTIVE: To observe the therapeutic effect and safety of perineal, crissal and progenital pruritus treated with acupuncture according to differentiation. METHODS: Self-control method was applied in these 32 cases. Changqiang (GV 1), Huiyin (CV 1), Qugu (CV 2), Sanyinjiao (SP 6) and Ashi points etc. were punctured as main points, and adjunct points were added according to differentiation: Taichong (LR 3) and Ququan (LR 8) etc. were added for wind and heat excess of liver meridian, Xuehai (SP 10) and Quchi (LI 11) etc. were added for blood deficiency and wind dryness. Itchiness, skin lesions sign scores and therapeutic effects were observed before and after treatment. RESULTS: The total scores of itchiness before and after treatment were 6.06 +/- 1.46 and 2.19 +/- 1.71 respectively, and the total scores of skin lesions sign were 4.38 +/- 2.21 and 1.50 +/- 1.44, indicating that the scores and the total scores of itchiness and skin lesions sign reduced obviously after treatment (P < 0.05, P < 0.01); the cured and markedly effective rate was 73.4% (11/15) for wind and heat excess of liver meridian, and 70.6% (12/17) for blood deficiency and wind dryness, presenting similar therapeutic effect (P > 0.05). Hematoma or ecchymosis appeared in 2 cases, and disappeared spontaneously after 2-3 days, without obvious adverse reaction. CONCLUSION: Simple perineal, crissal and progenital pruritus treated with acupuncture according to differentiation is effective, safe and applicable.

Practical method for detection and identification of Candida, Aspergillus, and Scedosporium spp. by use of rolling-circle amplification.

A sensitive rolling-circle amplification (RCA)-based method utilizing species-specific padlock probes targeted to the internal transcribed spacer 2 region of the fungal ribosomal DNA gene complex was developed. The assay was rapid (2 hours) and specific. Of 28 fungal isolates (16 of Candida, six of Aspergillus, and six of Scedosporium spp.), all were all identified correctly.

Expression of LL-37, human beta defensin-2, and CCR6 mRNA in patients with psoriasis vulgaris.

To investigate whether LL-37 and human beta defensin-2 (hBD-2) is related to the patients with psoriasis seldom having skin infections and explore the role of the two peptides and CCR6 (the receptor of hBD-2) in the pathogenesis of psoriasis, the expression levels of mRNA of LL-37, hBD-2, and CCR6 in skin lesions of patients with psoriasis vulgaris were detected by using RT-PCR. The results showed that the mRNA expression levels of the two peptides and CCR6 in psoriatic lesions all increased compared with the normal skin (P<0.001). It was suggested that up-regulated expression of LL-37 and hBD-2 might be the main reason that result in the the skin of patients with psoriasis being seldom infected, and the two peptides and CCR6 might play crucial roles in the pathogenesis of psoriasis.