Schistosoma japonicum extracellular vesicle proteins serve as effective biomarkers for diagnosing parasite infection.

Schistosoma species are the causative agent of schistosomiasis and shows worldwide distribution. There is a great need to develop a sensitive diagnostic approach for controlling the disease. Previously, we identified large numbers of Extracellular Vesicle (EV) proteins from Schistosoma japonicum (S. japonicum), but rarely these proteins have been evaluated for their diagnostic potential. In the present study, we performed bioinformatic analyses of S. japonicum identified EV-associated proteins from the previous study and then identified Schistosoma-specific proteins with potentially secreted capability. Among them, we selected SJCHGC02838 protein, SJCHGC05593 protein, SJCHGC05668 protein and a hypothetical protein (SJHYP) to evaluate their diagnostic potential for detecting S. japonicum infection. First, we determined the expression of these four proteins at the transcript levels using qRT-PCR and revealed that all these genes showed higher expression in adult stage. Then, we cloned the full-length cDNA for each protein into a prokaryotic expression vector and successfully generated the recombinant proteins. Upon the purification of recombinant proteins, we developed an indirect ELISA method to evaluate the diagnostic potential of these purified recombinant proteins. The results showed high sensitivity for detecting Schistosoma infection. Additionally, these proteins also displayed a good potential for detecting Schistosoma infection, especially SJCHGC05668 protein at an early stage. The diagnostic potentials of these recombinant proteins were further evaluated by Western blot and comparatively analyzed by our previously developed cfDNA methods.

Robust and adjustable dynamic scattering compensation for high-precision deep tissue optogenetics.

The development of high-precision optogenetics in deep tissue is limited due to the strong optical scattering induced by biological tissue. Although various wavefront shaping techniques have been developed to compensate the scattering, it is still a challenge to non-invasively characterize the dynamic scattered optical wavefront inside the living tissue. Here, we present a non-invasive scattering compensation system with fast multidither coherent optical adaptive technique (fCOAT), which allows the rapid wavefront correction and stable focusing in dynamic scattering medium. We achieve subcellular-resolution focusing through 500-µm-thickness brain slices, or even three pieces overlapped mouse skulls after just one iteration with a 589 nm CW laser. Further, focusing through dynamic scattering medium such as live rat ear is also successfully achieved. The formed focus can maintain longer than 60 s, which satisfies the requirements of stable optogenetics manipulation. Moreover, the focus size is adjustable from subcellular level to tens of microns to freely match the various manipulation targets. With the specially designed fCOAT system, we successfully achieve single-cellular optogenetic manipulation through the brain tissue, with a stimulation efficiency enhancement up to 300% compared with that of the speckle.

A multi-feature deep learning system to enhance glaucoma severity diagnosis with high accuracy and fast speed.

Glaucoma is the leading cause of irreversible blindness, and the early detection and timely treatment are essential for glaucoma management. However, due to the interindividual variability in the characteristics of glaucoma onset, a single feature is not yet sufficient for monitoring glaucoma progression in isolation. There is an urgent need to develop more comprehensive diagnostic methods with higher accuracy. In this study, we proposed a multi- feature deep learning (MFDL) system based on intraocular pressure (IOP), color fundus photograph (CFP) and visual field (VF) to classify the glaucoma into four severity levels. We designed a three-phase framework for glaucoma severity diagnosis from coarse to fine, which contains screening, detection and classification. We trained it on 6,131 samples from 3,324 patients and tested it on independent 240 samples from 185 patients. Our results show that MFDL achieved a higher accuracy of 0.842 (95 % CI, 0.795-0.888) than the direct four classification deep learning (DFC-DL, accuracy of 0.513 [0.449-0.576]), CFP-based single-feature deep learning (CFP-DL, accuracy of 0.483 [0.420-0.547]) and VF-based single-feature deep learning (VF-DL, accuracy of 0.725 [0.668-0.782]). Its performance was statistically significantly superior to that of 8 juniors. It also outperformed 3 seniors and 1 expert, and was comparable with 2 glaucoma experts (0.842 vs 0.854, p = 0.663; 0.842 vs 0.858, p = 0.580). With the assistance of MFDL, junior ophthalmologists achieved statistically significantly higher accuracy performance, with the increased accuracy ranged from 7.50 % to 17.9 %, and that of seniors and experts were 6.30 % to 7.50 % and 5.40 % to 7.50 %. The mean diagnosis time per patient of MFDL was 5.96 s. The proposed model can potentially assist ophthalmologists in efficient and accurate glaucoma diagnosis that could aid the clinical management of glaucoma.

Long-term microglial phase-specific dynamics during single vessel occlusion and recanalization.

Vascular occlusion leading to brain dysfunctions is usually considered evoking microglia-induced inflammation response. However, it remains unclear how microglia interact with blood vessels in the development of vascular occlusion-related brain disorders. Here, we illuminate long-term spatiotemporal dynamics of microglia during single vessel occlusion and recanalization. Microglia display remarkable response characteristics in different phases, including acute reaction, rapid diffusion, transition and chronic effect. Fibrinogen-induced microglial cluster promotes major histocompatibility complex II (MHCII) expression. Microglial soma represents a unique filament-shape migration and has slower motility compared to the immediate reaction of processes to occlusion. We capture proliferative microglia redistribute territory. Microglial cluster resolves gradually and microglia recover to resting state both in the morphology and function in the chronic effect phase. Therefore, our study offers a comprehensive analysis of spatiotemporal dynamics of microglia and potential mechanisms to both vessel occlusion and recanalization. Microglial phase-specific response suggests the morphological feature-oriented phased intervention would be an attractive option for vascular occlusion-related diseases treatments.

Cysteine and glycine-rich protein 2 promotes hypoxic pulmonary vascular smooth muscle cell proliferation through the Wnt3α-ß-catenin/lymphoid enhancer-binding factor 1 pathway.

Pulmonary hypertension (PH) is mainly characterized by abnormal pulmonary vascular hyperplasia and vascular remodeling, but its mechanism is complicated and currently unclear. Cysteine and glycine-rich protein 2 (Csrp2) has been reported to promote cell proliferation and migration, and affect cell cycle progression. As a new invasive actin-binding factor, Csrp2 increased the invasion and even metastasis of some cancer cells. It was associated with tumor recurrence and chemotherapy resistance. However, the role of Csrp2 in PH remains unknown. We found that Csrp2 expression was increased both in pulmonary arteries (PAs) and smooth muscle cells (PASMCs) in PH. Csrp2 enhanced PASMC proliferation and phenotypic transition. The Wnt3α-ß-catenin/lymphoid enhancer-binding factor 1 (LEF1) pathway is involved in cell proliferation and phenotypic transition regulated by Csrp2 expression. These results suggest that hypoxia downregulates YinYang-1 (YY1) and then increases Csrp2 expression. Increased Csrp2 promotes PASMC proliferation and phenotypic transition by activating the Wnt3α-ß-catenin/LEF1 pathways, which leads to pulmonary vascular remodeling and even provides a new theoretical basis for studying the pathogenesis and therapeutic targets of PH.

Raman-scattering-assistant large energy dissipative soliton and multicolor coherent noise-like pulse complex in an Yb-doped fiber laser.

In this Letter, we have demonstrated the generation of dissipative solitons (DSs) or multi-wavelength noise-like pulses (NLPs) directly from a common linear Yb-doped fiber laser in the presence of stimulated Raman scattering (SRS). For the DSs, the pulse energy of the solitons with a pulse width of 74.2 ps reaches 21.2 nJ. For the NLPs, the generation of the main NLP (1032 nm) together with the first-order Raman NLP (1080 nm) is realized. The narrow peak of the double-scale autocorrelation trace is characterized by quasi-periodic beat pulses with a pulse beating of 40.6 fs and a pulse separation of 79 fs, indicating that the generated solitons at dual wavelengths are mutually coherent. Furthermore, a three-color stable NLP complex with a broader spectrum is also obtained. The results contribute to an in-depth understanding of nonlinear dynamics and ultrafast physics.

Improved spatial resolution using focal modulation microscopy with a Tai Chi aperture.

Focal modulation microscopy (FMM) has gained significant interest in biological imaging. However, the spatial resolution and penetration depth limit the imaging quality of FMM due to the strong scattering background. Here, we introduce FMM with a Tai Chi aperture (TCFMM) based on diffraction theory to improve the spatial resolution. The results show that the transverse resolution is improved by 61.60% and 41.37% in two orthogonal directions, and the axial resolution is improved by 29.67%, compared with confocal microscopy (CM). The signal background ratio (SBR) of TCFMM is increased by 23.26% compared with CM and remains nearly the same compared with FMM using D-shape apertures (DFMM). These improvements in spatial resolution and SBR indicate that TCFMM has potential in deep tissue imaging.