Untrained neural network enabling fast and universal structured-illumination microscopy.

Structured-illumination microscopy (SIM) offers a twofold resolution enhancement beyond the optical diffraction limit. At present, SIM requires several raw structured-illumination (SI) frames to reconstruct a super-resolution (SR) image, especially the time-consuming reconstruction of speckle SIM, which requires hundreds of SI frames. Considering this, we herein propose an untrained structured-illumination reconstruction neural network (USRNN) with known illumination patterns to reduce the amount of raw data that is required for speckle SIM reconstruction by 20 times and thus improve its temporal resolution. Benefiting from the unsupervised optimizing strategy and CNNs' structure priors, the high-frequency information is obtained from the network without the requirement of datasets; as a result, a high-fidelity SR image with approximately twofold resolution enhancement can be reconstructed using five frames or less. Experiments on reconstructing non-biological and biological samples demonstrate the high-speed and high-universality capabilities of our method.

Effects of Insulin-degrading enzyme on the proliferation of swine Sertoli cells.

Sertoli cells, the only somatic cells in testis seminiferous tubules, provide a supporting microenvironment for male germ cells and play essential roles in spermatogenesis. The insulin-degrading enzyme (IDE), a ubiquitous zinc peptidase of the inverzincin family, plays crucial role in sperm production, as IDE-knockout mice presented decreased testis weight and impaired sperm viability and morphology. However, whether and how IDE affects swine Sertoli cell proliferation remains unclear. Thus, in the present study, we aimed to evaluate the effects of IDE on the proliferation of swine Sertoli cells, as well as its underlying molecular mechanism. After knocking down IDE expression with small interfering RNA transfection, we analyzed the proliferation of swine Sertoli cells as well as the expression of related regulatory factors (WT1, ERK, and AKT). The results showed that IDE knockdown promoted swine Sertoli cell proliferation and increased WT1 expression, possibly through activating ERK and AKT. Overall, our findings suggest that IDE may be involved in male reproduction by regulating Sertoli cell proliferation, which provides new information to better understand the regulatory mechanisms of swine Sertoli cells and improve the reproductive traits of male pigs.

An open, observational, three-arm clinical study of 2-3 cycles of treatment as neoadjuvant therapy in operable locally advanced non-small cell lung cancer: An interim analysis.

Background: An open, observational, three-arm clinical study aimed at investigating the efficacy of different neoadjuvant therapies (neoadjuvant immunotherapy with(out) chemotherapy, neoadjuvant chemotherapy, and neoadjuvant targeted therapy) in operable locally advanced non-small cell lung cancer (NSCLC) was conducted (NCT04197076). We report an interim analysis of 49 of 53 evaluable patients. Methods: This study was conducted at Shanghai Chest Hospital and included eligible NSCLC patients who were 18 years old and had clinical stage IIB-IIIB disease. All 49 patients had surgical resection within 4-6 weeks after 2-3 cycles of neoadjuvant treatment consisting of immunotherapy (24 patients), chemotherapy (16 patients), and a targeted therapy (9 patients) regimen starting on the first day of each 21-day cycle. Pathologic complete response (pCR) was evaluated as the primary endpoint. Major pathological response (MPR) and tumor regression rate (TRR) were also evaluated. Results: An improved pathologic complete response was achieved in the neoadjuvant immunotherapy arm compared with the neoadjuvant chemotherapy arm and neoadjuvant targeted therapy arm [20.8% (5/24) vs. 6.3% (1/16) vs. 0.0% (0/9); P = 0.089, 95% CI 0.138-0.151]. More importantly, we found that the curative effect of the neoadjuvant immunotherapy arm in pCR+MPR was better than that of the neoadjuvant chemotherapy arm and neoadjuvant targeted therapy arm [45.8% (11/24) vs. 18.8% (3/16) vs. 0.0% (0/9); P = 0.006, 95% confidence interval, 0.008-0.012]. Different neoadjuvant therapies had a statistically significant effect on postoperative pathological tumor downstaging (P = 0.017). Conclusions: Neoadjuvant immunotherapy was associated with a trend toward better pCR than the neoadjuvant chemotherapy arm and neoadjuvant targeted therapy. Curative effect (pCR + MPR) was significantly better with neoadjuvant immunotherapy (P = 0.006, 95% confidence interval, 0.008-0.012). Clinical Trial Registration: https://clinicaltrials.gov/ct2/show/NCT04197076?recrs=a&cond=NCT04197076&draw=2&rank=1.

Ring finger protein 6 enhances chemo-resistance by transcriptionally activating proliferating cell nuclear antigen expression and attenuating DNA damage in lung adenocarcinoma.

The E3 ubiquitin ligase RING finger protein 6 (RNF6) is elevated in several cancers, including prostate and colorectal cancers. Here, we extended the finding of elevated RNF6 expression levels and its association with poor prognosis in patients with lung adenocarcinoma (LUAD). Genome-wide RNA sequencing in H3255 cells with RNF6 knockdown, followed by analysis of differentially expressed genes using Clusters of Orthologous Groups and gene set enrichment analysis revealed aberrations in genes related to DNA repair, especially double-strand break (DSB) repair. RNF6 knockdown increased γH2AX foci, a biomarker for DSBs in H3255 and A549 LUAD cells, and enhanced DNA damage induced by chemotherapy in cisplatin-resistant A549/CDDP cells. In a series of experiments in cultured cells, as well as in nude mice carrying xenografts, RNF6 knockdown restored the sensitivity of A549/CDDP cells to cisplatin treatment. Mechanistically, RNA sequencing in RNF6-knockdown cells revealed the significant downregulation of proliferating cell nuclear antigen (PCNA), an oncogene that promotes DNA repair. Re-chromatin immunoprecipitation assay results suggested the formation of a RNF6-TCF4 complex that binds to the PCNA promoter to activate its transcription. Downregulation of RNF6 reduced TCF4 recruitment to PCNA promoters in H3255 and A549 cells, indicating that RNF6 regulates PCNA transcription to a certain extent by regulating TCF4 binding to PCNA promoters. The collective results implicate RNF6 overexpression as a molecular target in the management of cisplatin-resistant LUAD.

Total variation and spatial iteration-based 3D structured illumination microscopy.

Three-dimensional structured illumination microscopy (3D-SIM) plays an essential role in biological volumetric imaging with the capabilities of improving lateral and axial resolution. However, the traditional linear 3D algorithm is sensitive to noise and generates artifacts, while the low temporal resolution hinders live-cell imaging. In this paper, we propose a novel 3D-SIM algorithm based on total variation (TV) and fast iterative shrinkage threshold algorithm (FISTA), termed TV-FISTA-SIM. Compared to conventional algorithms, TV-FISTA-SIM achieves higher reconstruction fidelity with the least artifacts, even when the signal-to-noise ratio (SNR) is as low as 5 dB, and a faster reconstruction rate. Through simulation, we have verified that TV-FISTA-SIM can effectively reduce the amount of required data with less deterioration. Moreover, we demonstrate TV-FISTA-SIM for high-quality multi-color 3D super-resolution imaging, which can be potentially applied to live-cell imaging applications.

3D super-resolution microscopy based on nonlinear gradient descent structured illumination.

Three-dimensional structured illumination microscopy (3D-SIM) is an essential tool for volumetric fluorescence imaging, which improves both axial and lateral resolution by down-modulating high-frequency information of the sample into the passband of optical transfer function (OTF). And when combining with the 4Pi structure, the performance of 3D-SIM can be further improved. The reconstruction results of generally used linear 3D algorithm, however, are lack of high-fidelity and proneess to generate artifacts. In this paper, we proposed a novel iterative algorithm based on gradient descent combined with a nonlinear optimizer, which can be applied to all 3D-SIM setups (including I5S setup). We verified through simulation that the proposed solution, termed as nonlinear gradient descent structured illumination microscopy (NGD-SIM), achieves more fidelity results which can reach the limitation of theoretical resolution improvement of SIM. Moreover, it can be firmly validated on simulation that this algorithm can effectively reduce the amount of raw data in the case of sinusoidal-pattern illumination, i.e., the algorithm doesn't need five-step phase shifting; data with any number of phases can theoretically be reconstructed. Our method also provides the possibility to extend the application of sinusoidal-pattern illumination to any kind of interference fringe, which is generated by diversified types of illumination mode.

Decision-Making and Multisensory Combination Under Time Stress.

Using a video game platform, we examined how vision-based decision making was affected by a concurrent, potentially conflicting auditory stimulus. Electroencephalographic responses showed that by 150 milliseconds of stimulus onset, the brain had detected the conflict between visual and auditory stimuli. Systematically reducing the intertrial interval (ITI), which subjects described as stressful, undermined decision making. Subjects' arterial pulse variance decreased along with ITI, signaling increased parasympathetic influence on the heart. When successive trials required a shift in processing mode, short ITIs significantly boosted one trial's influence on the next, suggesting that stress reduces cognitive flexibility. Finally, our study demonstrates the heart's and the brain's important influence on decision making.

Mini-patient-derived xenograft assay based on microfluidic technology promises to be an effective tool for screening individualized chemotherapy regimens for advanced non-small cell lung cancer.

Patient-derived xenograft (PDX) assay has been widely used in preclinical research in patients with multidrug-resistant lung cancer. One hundred patients with non-small cell lung cancer (NSCLC) were divided into MiniPDX group and conventional group, with 50 cases in each group. The MiniPDX assay was established by enriching high-purity tumor cells using microfluidic technology to detect the drug sensitivity of NSCLC cells. All patients underwent conventional computed tomography (CT) scans of lung and mediastinum at baseline and during follow-up. Kaplan-Meier method was used to compare the overall survival and progression-free survival of two groups. The sensitivity of the same drug in different tumor xenograft varied greatly. The overall survival, progression-free survival, and clinical benefit rate of patients in the MiniPDX-guided chemotherapy group were significantly longer than those in the conventional chemotherapy group. MiniPDX assay may be an effective tool for screening chemotherapy regimens in NSCLC patients.

Comparative Efficacy and Safety of Immunotherapy Alone and in Combination With Chemotherapy for Advanced Non-small Cell Lung Cancer.

There is a lack of direct cross-comparison studies in clinical trials between immunotherapy alone and combination treatment, especially in Non-Small Cell Lung Cancer (NSCLC) patients with high PD-L1 expression. To determine if anti-PD-(L)1 antibody combined with chemotherapy is more efficient than immune checkpoint inhibitor (ICI) monotherapy for advanced NSCLC patients in the real-world data. We retrospectively collected 325 patients with advanced NSCLC treated with ICI alone with or without chemotherapy from 11th July 2016 to 26th May 2020 to investigate which treatment scenario is the most efficient, and how clinical factors impact response. Patients with advanced NSCLC were treated with ICI monotherapy (178/325, 54.8%) or in combination with chemotherapy (147/325, 45.2%). The objective response rate and disease control rate were higher in the combination group than the monotherapy group. Patients (including those with distant metastasis) treated with chemo-immunotherapy were associated with a significantly longer median PFS and OS compared with the monotherapy group, irrespective of the PD-L1 expression level and previous treatment lines. No significant increase in the risk of immune-related adverse events (irAEs) was found after combination with chemotherapy (50.6 vs. 57.8%). IrAEs predicted better PFS of immunotherapy in the monotherapy group, especially for patients with late irAEs (after ≥4 cycles). Collectively, we demonstrated that ICI monotherapy plus chemotherapy might have better anti-tumor activity and an acceptable side-effect profile regardless of PD-L1 level or previous treatment lines. Both regimens were well-tolerated and cost-effective, the more efficient is usually recommended.

Histone 3 lysine-27 demethylase KDM6A coordinates with KMT2B to play an oncogenic role in NSCLC by regulating H3K4me3.

Aberrations in epigenetic modulation dysregulate transcription, playing a critical role in the developmental process of tumors, including lung cancer. Aberrant levels of the histone 3 lysine-27 demethylase KDM6A have been found in cancer and are either positively or negatively associated with tumorigenesis and prognosis. However, the clinical relevance and functional role of KDM6A in lung cancer is largely unknown. We found that KDM6A protein expression was higher in NSCLC tissues than in the corresponding paracancer tissues and that high KDM6A expression was associated with poor patient prognosis. Furthermore, KDM6A knockdown in NSCLC cell lines markedly inhibited the tumorigenic phenotype both in vitro and in vivo. Mechanistically, KDM6A colocalized and cooperated with KMT2B to reprogram the transcriptional network via regulating the cancer pathway, in which abnormal activation of the Wnt pathway is the dominant factor. Interestingly, in NSCLC cell lines, H3K4me3 but not H3K27me2/3 or H3K4me1/2 was markedly altered upon KDM6A or KMT2B knockdown, indicating that KDM6A may act independently of H3K27 demethylases in NSCLC. Taken together, these results indicated that KDM6A or KMT2B may be a prognostic biomarker and promising therapeutic target in NSCLC.

Survival comparison of Three histological subtypes of lung squamous cell carcinoma: A population-based propensity score matching analysis.

OBJECTIVES: This study is aimed to analyze the survival differences among patients with lung basaloid squamous cell carcinoma (BSCC), keratinizing squamous cell carcinoma (KSCC), and nonkeratinizing squamous cell carcinoma (NKSCC), and explore the prognostic factors of patients with lung BSCC. MATERIALS AND METHODS: We searched the information of 4743 patients with lung SCC between 2005 and 2014 from the SEER database. Propensity score matching (PSM) was used to adjust confounding factors. The overall survival (OS), cancer-specific survival (CSS), and cumulative incidence of cancer-specific mortality (CSM) were estimated with a comparative analysis. A competing risks regression model was conducted to identify the prognostic factors of lung BSCC. RESULTS: After PSM, patients with lung BSCC had a higher CSS rate than those with lung KSCC or NKSCC (5-year CSS rate: 50.4 % vs. 37.7 % vs. 38.5 %, p = 0.033 and p = 0.033). The cumulative incidence of CSM was lower for patients with lung BSCC than those with lung KSCC or NKSCC (5-year CSM rate: 46.4 % vs. 56.9 % vs. 56.4 %, p = 0.046 and p = 0.042), which were similar to the results before PSM. As for patients with lung KSCC and NKSCC, there was no survival differences between them (5-year CSS rate: 37.7 % vs. 38.5 %, p = 0.997). The competing risks regression analysis showed that T stage, N stage, M stage and surgery were independent prognostic factors for patients with lung BSCC (all p < 0.05). CONCLUSIONS: Patients with lung BSCC had a better survival than those with lung KSCC or NKSCC, while no survival differences were found between lung KSCC and NKSCC. T stage, N stage, M stage and surgery were independent prognostic factors for patients with lung BSCC.

Cross-Sectional Associations between Dietary Fat-Related Behaviors and Continuous Metabolic Syndrome Score among Young Australian Adults.

Dietary guidelines recommend removing visible fat from meat, choosing low-fat options and cooking with oil instead of butter. This study examined cross-sectional associations between fat-related eating behaviors and a continuous metabolic syndrome (cMetSyn) score among young adults. During 2004⁻2006, 2071 participants aged 26⁻36 years reported how often they trimmed fat from meat, consumed low-fat dairy products and used different types of fat for cooking. A fasting blood sample was collected. Blood pressure, weight and height were measured. To create the cMetSyn score, sex-specific principal component analysis was applied to normalized risk factors of the harmonized definition of metabolic syndrome. Higher score indicates higher risk. For each behavior, differences in mean cMetSyn score were calculated using linear regression adjusted for confounders. Analyses were stratified by weight status (Body mass index (BMI) < 25 kg/m² or ≥25 kg/m²). Mean cMetSyn score was positively associated with consumption of low-fat oily dressing (PTrend = 0.013) among participants who were healthy weight and frequency of using canola/sunflower oil for cooking (PTrend = 0.008) among participants who were overweight/obese. Trimming fat from meat, cooking with olive oil, cooking with butter, and consuming low-fat dairy products were not associated with cMetSyn score. Among young adults, following fat-related dietary recommendations tended to not be associated with metabolic risk.

Catching Audiovisual Interactions With a First-Person Fisherman Video Game.

The human brain is excellent at integrating information from different sources across multiple sensory modalities. To examine one particularly important form of multisensory interaction, we manipulated the temporal correlation between visual and auditory stimuli in a first-person fisherman video game. Subjects saw rapidly swimming fish whose size oscillated, either at 6 or 8 Hz. Subjects categorized each fish according to its rate of size oscillation, while trying to ignore a concurrent broadband sound seemingly emitted by the fish. In three experiments, categorization was faster and more accurate when the rate at which a fish oscillated in size matched the rate at which the accompanying, task-irrelevant sound was amplitude modulated. Control conditions showed that the difference between responses to matched and mismatched audiovisual signals reflected a performance gain in the matched condition, rather than a cost from the mismatched condition. The performance advantage with matched audiovisual signals was remarkably robust over changes in task demands between experiments. Performance with matched or unmatched audiovisual signals improved over successive trials at about the same rate, emblematic of perceptual learning in which visual oscillation rate becomes more discriminable with experience. Finally, analysis at the level of individual subjects' performance pointed to differences in the rates at which subjects can extract information from audiovisual stimuli.

Bi-directional audiovisual influences on temporal modulation discrimination.

Cross-modal interactions of auditory and visual temporal modulation were examined in a game-like experimental framework. Participants observed an audiovisual stimulus (an animated, sound-emitting fish) whose sound intensity and/or visual size oscillated sinusoidally at either 6 or 7 Hz. Participants made speeded judgments about the modulation rate in either the auditory or visual modality while doing their best to ignore information from the other modality. Modulation rate in the task-irrelevant modality matched the modulation rate in the task-relevant modality (congruent conditions), was at the other rate (incongruent conditions), or had no modulation (unmodulated conditions). Both performance accuracy and parameter estimates from drift-diffusion decision modeling indicated that (1) the presence of temporal modulation in both modalities, regardless of whether modulations were matched or mismatched in rate, resulted in audiovisual interactions; (2) congruence in audiovisual temporal modulation resulted in more reliable information processing; and (3) the effects of congruence appeared to be stronger when judging visual modulation rates (i.e., audition influencing vision), than when judging auditory modulation rates (i.e., vision influencing audition). The results demonstrate that audiovisual interactions from temporal modulations are bi-directional in nature, but with potential asymmetries in the size of the effect in each direction.

Policing Fish at Boston's Museum of Science: Studying Audiovisual Interaction in the Wild.

Boston's Museum of Science supports researchers whose projects advance science and provide educational opportunities to the Museum's visitors. For our project, 60 visitors to the Museum played "Fish Police!!," a video game that examines audiovisual integration, including the ability to ignore irrelevant sensory information. Players, who ranged in age from 6 to 82 years, made speeded responses to computer-generated fish that swam rapidly across a tablet display. Responses were to be based solely on the rate (6 or 8 Hz) at which a fish's size modulated, sinusoidally growing and shrinking. Accompanying each fish was a task-irrelevant broadband sound, amplitude modulated at either 6 or 8 Hz. The rates of visual and auditory modulation were either Congruent (both 6 Hz or 8 Hz) or Incongruent (6 and 8 or 8 and 6 Hz). Despite being instructed to ignore the sound, players of all ages responded more accurately and faster when a fish's auditory and visual signatures were Congruent. In a controlled laboratory setting, a related task produced comparable results, demonstrating the robustness of the audiovisual interaction reported here. Some suggestions are made for conducting research in public settings.

Factors contributing to de qi in acupuncture randomized clinical trials.

De qi is a core concept of acupuncture and is necessary to produce therapeutic effect. In 2010, de qi has been received as a term in the official extension of the CONSORT Statement. However, there are few articles that discuss which factors have influences on obtaining de qi in clinical trials. This paper aims to explore these factors and give advice on trial design in order to optimize de qi in acupuncture RCTs.

A match made by modafinil: probability matching in choice decisions and spatial attention.

When predicting where a target or reward will be, participants tend to choose each location commensurate with the true underlying probability (i.e., probability match). The strategy of probability matching involves independent sampling of high and low probability locations on separate trials. In contrast, models of probabilistic spatial attention hypothesize that on any given trial attention will either be weighted toward the high probability location or be distributed equally across all locations. Thus, the strategies of probabilistic sampling by choice decisions and spatial attention appear to differ with regard to low-probability events. This distinction is somewhat surprising because similar brain mechanisms (e.g., pFC-mediated cognitive control) are thought to be important in both functions. Thus, the goal of the current study was to examine the relationship between choice decisions and attentional selection within single trials to test for any strategic differences, then to determine whether that relationship is malleable to manipulations of catecholamine-modulated cognitive control with the drug modafinil. Our results demonstrate that spatial attention and choice decisions followed different strategies of probabilistic information selection on placebo, but that modafinil brought the pattern of spatial attention into alignment with that of predictive choices. Modafinil also produced earlier learning of the probability distribution. Together, these results suggest that enhancing cognitive control mechanisms (e.g., through prefrontal cortical function) leads spatial attention to follow choice decisions in selecting information according to rule-based expectations.