Discovery of the Clinical Candidate Sonrotoclax (BGB-11417), a Highly Potent and Selective Inhibitor for Both WT and G101V Mutant Bcl-2.

The approval of venetoclax, a B-cell lymphoma-2 (Bcl-2) selective inhibitor, for the treatment of chronic lymphocytic leukemia demonstrated that the antiapoptotic protein Bcl-2 is a druggable target for B-cell malignancies. However, venetoclax's limited potency cannot produce a strong, durable clinical benefit in other Bcl-2-mediated malignancies (e.g., diffuse large B-cell lymphomas) and multiple recurrent Bcl-2 mutations (e.g., G101V) have been reported to mediate resistance to venetoclax after long-term treatment. Herein, we described novel Bcl-2 inhibitors with increased potency for both wild-type (WT) and mutant Bcl-2. Comprehensive structure optimization led to the clinical candidate BGB-11417 (compound 12e, sonrotoclax), which exhibits strong in vitro and in vivo inhibitory activity against both WT Bcl-2 and the G101V mutant, as well as excellent selectivity over Bcl-xL without obvious cytochrome P450 inhibition. Currently, BGB-11417 is undergoing phase II/III clinical assessments as monotherapy and combination treatment.

Development and internal validation of a nomogram to predict temporary acute agitated delirium after surgery for chronic subdural hematoma in elderly patients: an analysis of the clinical database.

Background: This study aimed to develop a nomogram for predicting temporary acute agitated delirium after surgery in patients with chronic subdural hematoma (CSH) without neurological compromise and hospitalized in the neurosurgery. Methods: We included 289 patients with chronic subdural hematoma (CSH) from the medical information system of Yuebei People's Hospital of Shaoguan City, Guangdong Province, and collected 16 clinical indicators within 24 h of admission. We used the least absolute shrinkage and selection operator (LASSO) regression to identify risk factors. We established a multivariate logistic regression model and constructed a nomogram. We performed internal validation by 1,000 bootstrap samples; we plotted a receiver operating curve (ROC) and calculated the area under the curve (AUC), sensitivity, and specificity. We also evaluated the calibration of our model by the calibration curve and the Hosmer-Lemeshow goodness-of-fit test (HL test). We performed a decision curve analysis (DCA) and a clinical impact curve (CIC) to assess the net clinical benefit of our model. Results: The nomogram included alcoholism history, hepatic insufficiency, verbal rating scale for postoperative pain (VRS), pre-hospital modified Rankin Scale (mRS), and preoperative hematoma thickness as predictors. Our model showed satisfactory diagnostic performance with an AUC value of 0.8474 in the validation set. The calibration curve and the HL test showed good agreement between predicted and observed outcomes (p = 0.9288). The DCA and CIC showed that our model had a high predictive ability for the occurrence of postoperative delirium in patients with CSDH. Conclusion: We identified alcoholism, liver dysfunction, pre-hospital mRS, preoperative hematoma thickness, and postoperative VRS pain as predictors of postoperative delirium in chronic subdural hematoma patients. We developed and validated a multivariate logistic regression model and a nomogram.

Correction to "Solvent-Free Nonthermal Destruction of PFAS Chemicals and PFAS in Sediment by Piezoelectric Ball Milling".

[This corrects the article DOI: 10.1021/acs.estlett.2c00902.].

ZC3H13 promotes ITGA6 m6A modification for chronic obstructive pulmonary disease progression.

Chronic obstructive pulmonary disease (COPD) is potentially fatal, and as society ages, its effects on human health are predicted to deteriorate. The potential function of m6A modifications within COPD has become a hot topic recently. This study was conducted to clarify the function and related mechanisms of the m6A methylation transferase ZC3H13 in COPD. The expression of m6A-associated protease and ITGA6 in COPD tissues was assessed using GEO data, qRT-PCR, and western blot. COPD models in cells and mice were established through cigarette smoke extract (CSE) and smoke exposure. Inflammatory marker levels were measured by ELISA, apoptosis by flow cytometry, and mRNA stability with Actinomycin D assay. m6A modification levels were checked by MeRIP-PCR. HE and Masson staining evaluated lung pathology, and alveolar lavage fluid analysis included total cell count and Giemsa staining. ZC3H13 and METTL3 were differentially expressed m6A regulators in COPD, with ZC3H13 being more significantly upregulated. Further analysis revealed the ZC3H13 expression-related differentially expressed genes (DEGs) functions were enriched in the immunoinflammatory pathway, indicating ZC3H13's involvement in COPD pathogenesis through inflammation, and immune responses. Knockdown studies in cellular and mouse models demonstrated ZC3H13's role in exacerbating COPD symptoms, including inflammation, apoptosis, and EMT, and its suppression led to significant improvements. The identification of ITGA6 as a target gene further elucidated the mechanism, showing that ZC3H13 enhances ITGA6 expression and mRNA stability through m6A modification, influencing bronchial epithelial cell inflammation and fibrosis. In conclusion, targeting ZC3H13/ITGA6 could be an underlying therapeutic approach for treating COPD.

Prescribed-time containment control of multi-agent systems subject to collision avoidance and connectivity maintenance.

In this paper, the problem of prescribed-time containment control for a second-order multiple leader-follower systems (MLFSs) is studied, in where both collision avoidance and connectivity maintenance of the agents are considered. Firstly, an effective exponential potential field function (EAPF) with constraints based on the estimated distance is designed to achieve collision resistance and connectivity preservation of the agents at a prescribed-time. Secondly, an estimator-based distributed control protocol is proposed, which drives the agents to achieve containment control in a cooperative manner at a prescribed-time. Furthermore, a novel distributed control protocol containing a collision avoidance term and a containment control term is addressed as well, which enables all followers to complete collision avoidance and connectivity maintenance in any prescribed-time and enter the leaders' convex packet. Finally, the stability of the system is technically analyzed by using Lyapunov theory, and the effectiveness of the presented strategies is verified by several simulations.

Leader-Following Synchronization Control of Multiagent Systems Under Hybrid Cyber Attacks via Impulsive Control Based on Topology Switching.

This article investigates the leader-following synchronization problem of multiagent systems (MASs) under hybrid cyber attacks, which refers to deception attacks and multichannel independent denial-of-service (DoS) attacks in communication channels. In order to achieve the secure control of MASs under hybrid cyber attacks, a novel impulsive control method based on topology switching is proposed, and a new algorithm for determining impulsive instants is designed. In addition, the cooperative-competitive relationship between agents is also considered, which is more in line with reality. Sufficient conditions for ensuring secure control of MASs and a parametric upper bound on the error vector norm between the agents and the leader are obtained. Finally, the numerical simulation verified the effectiveness of the proposed algorithm.

Extending the Grading of Recommendations Assessment, Development and Evaluation (GRADE) in Traditional Chinese Medicine (TCM): The GRADE-TCM.

AIM: To extend and form the "Grading of Recommendations Assessment, Development and Evaluation in Traditional Chinese Medicine" (GRADE-TCM). METHODS: Methodologies were systematically reviewed and analyzed concerning evidence-based TCM guidelines worldwide. A survey questionnaire was developed based on the literature review and open-end expert interviews. Then, we performed expert consensus, discussion meeting, opinion collection, external examination, and the GRADE-TCM was formed eventually. RESULTS: 265 Chinese and English TCM guidelines were included and analyzed. Five experts completed the open-end interviews. Ten methodological entries were summarized, screened and selected. One round of consensus was conducted, including a total of 22 experts and 220 valid questionnaire entries, concerning 1) selection of the GRADE, 2) GRADE-TCM upgrading criteria, 3) GRADE-TCM evaluation standard, 4) principles of consensus and recommendation, and 5) presentation of the GRADE-TCM and recommendation. Finally, consensus was reached on the above 10 entries, and the results were of high importance (with voting percentages ranging from 50 % to 81.82 % for "very important" rating) and strong reliability (with the Cr ranging from 0.93 to 0.99). Expert discussion meeting (with 40 experts), opinion collection (in two online platforms) and external examination (with 14 third-party experts) were conducted, and the GRADE-TCM was established eventually. CONCLUSION: GRADE-TCM provides a new extended evidence-based evaluation standard for TCM guidelines. In GRADE-TCM, international evidence-based norms, characteristics of TCM intervention, and inheritance of TCM culture were combined organically and followed. This is helpful for localization of the GRADE in TCM and internationalization of TCM guidelines.

Sonrotoclax overcomes BCL2 G101V mutation-induced venetoclax resistance in preclinical models of hematologic malignancy.

ABSTRACT: Venetoclax, the first-generation inhibitor of the apoptosis regulator B-cell lymphoma 2 (BCL2), disrupts the interaction between BCL2 and proapoptotic proteins, promoting the apoptosis in malignant cells. Venetoclax is the mainstay of therapy for relapsed chronic lymphocytic leukemia and is under investigation in multiple clinical trials for the treatment of various cancers. Although venetoclax treatment can result in high rates of durable remission, relapse has been widely observed, indicating the emergence of drug resistance. The G101V mutation in BCL2 is frequently observed in patients who relapsed treated with venetoclax and sufficient to confer resistance to venetoclax by interfering with compound binding. Therefore, the development of next-generation BCL2 inhibitors to overcome drug resistance is urgently needed. In this study, we discovered that sonrotoclax, a potent and selective BCL2 inhibitor, demonstrates stronger cytotoxic activity in various hematologic cancer cells and more profound tumor growth inhibition in multiple hematologic tumor models than venetoclax. Notably, sonrotoclax effectively inhibits venetoclax-resistant BCL2 variants, such as G101V. The crystal structures of wild-type BCL2/BCL2 G101V in complex with sonrotoclax revealed that sonrotoclax adopts a novel binding mode within the P2 pocket of BCL2 and could explain why sonrotoclax maintains stronger potency than venetoclax against the G101V mutant. In summary, sonrotoclax emerges as a potential second-generation BCL2 inhibitor for the treatment of hematologic malignancies with the potential to overcome BCL2 mutation-induced venetoclax resistance. Sonrotoclax is currently under investigation in multiple clinical trials.

The predictive role of the nomogram based on clinical characteristics and thromboelastography markers for rebleeding after hypertensive intracerebral hemorrhage.

Objectives: Rebleeding after hypertensive intracerebral hemorrhage is a common and serious postoperative complication in neurosurgery, with high mortality and mental disability rates. The aim of this study was to establish a nomogram to analyze the role of thromboelastography in predicting rebleeding after hypertensive intracerebral hemorrhage. Basic methods: We selected 375 patients with hypertensive intracerebral hemorrhage who underwent surgical treatment in Yuebei People's Hospital of Shaoguan City, Guangdong Province from May 2018 to August 2022, and retrospectively analyzed the relevant data of hypertensive intracerebral hemorrhage patients (including general data and clinical thromboelastography data), and analyzed the factors and thromboelastography parameters that affect rebleeding after surgery (45 cases, defined as re-examination of head CT within 72 h after surgery showed that the hematoma in the surgical area exceeded 20 ml). Main results: Time from symptom onset to surgery, taking antiplatelet drugs, taking anticoagulant drugs, diabetes mellitus, difficulty in hemostasis during surgery, R value and EPL value in thromboelastography were risk factors for rebleeding after hypertensive intracerebral hemorrhage (P < 0.05). Logistic regression was used to determine the independent risk factors, and based on these risk factors, a nomogram was established and internally validated using a bootstrap method. ROC curve analysis showed that the nomogram model had high diagnostic value for rebleeding after hypertensive intracerebral hemorrhage, with AUC of 0.7314. The calibration curve of the nomogram showed good consistency between the predicted probabilities and the observed values. The decision curve analysis and clinical impact curve also revealed the potential clinical usefulness of the nomogram. Conclusions: The nomogram based on clinical characteristics and thromboelastography markers may be useful for predicting rebleeding after hypertensive intracerebral hemorrhage.

Longitudinal Changes in Chinese Prosociality.

This article presents three studies using data from the World Values Survey, 128 published studies, and China Family Panel Studies to comprehensively examine the longitudinal dynamics of Chinese prosociality, encompassing prosocial attitudes, tendencies, and behaviors, with the overarching goal of shedding light on the evolving nature of prosociality in the Chinese context. These studies reveal a consistent pattern, illustrating a decline followed by a resurgence in all three aspects, with a nadir around 2014. In addition, the study investigates the intricate relationship between economic inequality, prosocial behavior, and prosocial attitudes. The findings suggest that while economic inequality significantly relates to prosocial behavior, it does not entirely explain its fluctuations. Prosocial attitudes partially mediate the connection between economic inequality and prosocial behavior. These insights suggest that addressing inequality could contribute to a more conducive social environment for societal-level prosociality. However, further research is imperative to explore additional determinants of prosociality shifts.

IKZF4/NONO-RAB11FIP3 axis promotes immune evasion in gastric cancer via facilitating PD-L1 endosome recycling.

As an immune checkpoint protein expressed by diverse cancer cells, programmed death ligand 1 (PD-L1) facilitates immune evasion by interacting with programmed cell death-1 (PD-1) on T cells. Despite the clinical benefits observed in various cancer types, strategies targeting PD-1/PD-L1 have demonstrated limited efficacy in gastric cancer (GC). Furthermore, the regulation of PD-L1, especially at post-translational modification levels, remains largely unknown. Therefore, it is crucial to elucidate the mechanisms governing PD-L1 expression to enhance anti-tumor immunity. In this study, we have identified that IKAROS family zinc finger 4 (IKZF4) and Non-POU domain-containing octamer-binding (NONO) synergistically regulate and enhance the expression of RAB11 family-interacting protein 3 (RAB11FIP3) in GC. The IKZF4/NONO-RAB11FIP3 axis facilitates the endosomal recycling of PD-L1, particularly on the cell membrane of GC cells. Moreover, overexpression of RAB11FIP3 mitigates the hypo-expression of PD-L1 protein resulting from IKZF4 or NONO deletion. Functionally, the silencing of RAB11FIP3 or IKZF4 promotes T cell proliferation, and enhances T-cell cytotoxicity towards GC cells in vitro, which further inhibits tumor immune evasion in mice via increasing the infiltration of CD8+ T cells into the tumor microenvironment (TME) to suppress GC progression. Our study suggests that the IKZF4/NONO-RAB11FIP3 axis promotes immune evasion by facilitating PD-L1 endosome recycling, thus presenting a potential therapeutic target for GC treatment.

Comparative Assessment of APTT Reagents for Evaluating Anticoagulant Sensitivity of Fucosylated Glycosaminoglycans (FGs) Derived from Sea Cucumbers.

Fucosylated glycosaminoglycans (FGs) derived from sea cucumbers exhibit potent intrinsic Xase (iXase) inhibition, anticoagulation, and antithrombosis. Plasma activated partial thromboplastin time (APTT), a widely used screening test worldwide, is crucial for evaluating anticoagulant efficacy. However, the applicability of these commercially available APTT reagents for assessing anticoagulation of FGs remains unreported. In this study, we investigated the disparity between ellagic acid and colloidal silica APTT reagents in evaluating anticoagulation of dHG-5 and dHLFG-4, two depolymerized FGs, and elucidated the underlying rationale. The results demonstrated that dHG-5 and dHLFG-4 exhibited heightened sensitivity to the ellagic acid APTT reagent both in vitro and in vivo, and did not significantly affect the activation of APTT reagents for plasma. In addition, both ellagic acid and colloidal silica APTT reagents inhibited the anti-iXase of dHG-5 and dHLFG-4, and the inhibition of the ellagic acid APTT reagent was less pronounced compared to the colloidal silica APTT reagent. These findings suggest that the reduced impact of the ellagic acid APTT reagent on the anti-iXase activity of dHG-5 and dHLFG-4 is responsible for the increased sensitivity in plasma APTT analysis. This study offers valuable insights into the characteristics of two APTT reagents applied for assessing the anticoagulant activity of FG-related compounds.

Quantitative simulation and verification of the tourism economic resilience in urban agglomerations.

The concept of tourism economic resilience emphasizes the sustainable development level of tourism economy under uncertainty and risk. Focusing on urban agglomerations, this study aims to describe how the tourism economic resilience is developing, explore whether the resilience level is enhanced with urban agglomerations and whether spatial elements affect resilience levels. With the combination of the aggregation and diffusion effects and crowding effects of regional development, the study uses a combination of dynamic evaluation method, spatial kernel density, and mathematical models of urban agglomeration development to quantitatively analyze the spatiotemporal dynamic evolution of tourism economic resilience from 2006 to 2019, simulates and verifies its development patterns. The conclusions show that: (1) The tourism economic resilience in urban agglomerations is closely related to regional development and prosperity; (2) The development of tourism economic resilience also follows the spatial economic development pattern which moves towards equilibrium in aggregation process; (3) The tourism economic resilience of urban agglomerations has a fluctuation climbing node, generally presents as a wave-like upward trend with fluctuations and stages; (4) The evolutionary trend of tourism economic resilience in urban agglomerations presents as a slight wave-like upward curve that changes with time and co-opetition.

Study on the performance of a new type of combined packing biofilm reactor treating wastewater.

The present work investigates the performance of a biofilm reactor filled with a new type of combined packing used to treat wastewater and explores a new technology approach for the application of coral sand and waste non-woven fabric. The combined packing was made of coral sand and waste non-woven fabric, which was used as a biofilm carrier to treat sewage. The experimental results showed that the removal efficiencies of COD, NH4+-N and TN in the biofilm reactor containing the combined packing were 92.9%, 72.9% and 63.2%, respectively. The maximum removal efficiencies of COD, NH4+-N and TN in the biofilm reactor containing single packing were 89.0%, 63.4% and 55.2%, respectively. The properties of the combined packing were characterized by Fourier Transform Infrared (FTIR), specific surface area, SEM and dehydrogenase activity. Infrared analysis showed that there were hydroxyl, carboxyl and carbonyl groups on the surface of coral sand and non-woven fabric which were beneficial for biofilm growth and wastewater treatment. The large pores in the interior of coral sand and non-woven fabric could provide a comfortable environment for microbes to grow and reproduce. The dehydrogenase activity of the biofilm on the surface of coral sand in the third biofilm reactor was 49.91 µgTF·g-1·h-1, which was significantly higher than that of the other two biofilm reactors. The new type of combined packing is suitable for biofilm carriers with low cost, which can be applied to actual sewage treatment projects. This study provides a reference for the practical application of the technique.

Comprehensive analysis of ferritinophagy-related genes and immune infiltration landscape in diabetic retinopathy.

Background: Diabetic retinopathy (DR) is deemed a microangiopathy and neurodegenerative disorder, which is a primary reason of visual impairment in the world. Ferritinophagy is a critical regulator of ferroptosis and has a vital part in the etiopathogenesis of DR. Nevertheless, its molecular mechanism in DR remains to be expounded. Methods: The GSE146615 dataset was adopted to identify ferritinophagy-related differentially expressed genes (FRDEGs). The interactions and biological functions of the genes were described by means of functional enrichment analysis (FEA). The enriched gene sets were analyzed utilizing gene set enrichment analysis (GSEA) and gene set variation analysis (GSVA). Identification of hub genes was performed utilizing protein-protein interaction (PPI) analysis. mRNA-miRNA, mRNA-transcription factors (TF), mRNA-drugs, mRNA-RNA-binding proteins (RBP) interaction networks were constructed. In addition, datasets GSE60436 and GSE94019 were utilized for validation. The diagnostic performance of FRDEGs was assessed by means of receiver-operating characteristic curve monofactor analysis, followed by immune infiltration analysis. Lastly, quantitative real-time polymerase chain reaction (qRT-PCR) was implemented to analyze the validation of genes. Results: In total, the identification of eight FRDEGs was completed utilizing differential expression analysis. FEA mainly implicated the autophagy of mitochondrion, mitochondrion disassembly, autophagosome assembly, and organization pathways. GSEA and GSVA mainly implicated the interferon alpha response, ultraviolet response up, interferon gamma response, apical junction, pical surface, and allograft rejection pathways. BECN1 and HERC2 displayed high diagnostic accuracies in validation sets. Immune infiltration analysis revealed that several immune cells related to ferritinophagy may be play potential roles in DR. Finally, qRT-PCR was utilized to validate the upregulated expression of BECN1 as well as the downregulated expression of BCAT2 and ATG7 in the DR model. Conclusion: BECN1, HERC2, ATG7, and BCAT2 act as potential biomarkers for DR and might regulate ferritinophagy and the immune microenvironment to influence its development and progression. This research can provide new insights into pathogenesis of DR related to ferritinophagy.

Spatial dynamics and influencing factors of carbon rebound effect in tourism transport: Evidence from the Yangtze-river delta urban agglomeration.

Economic efficiency gains in tourism are considered a crucial approach to reducing carbon emissions in the tourism sector, especially in tourism transport. However, as a significant source of carbon emissions from tourism activities, the total carbon emissions from tourism transport have not decreased proportionally to the reduction in the intensity, despite China's overall improvement in the tourism economic efficiency. This phenomenon is commonly known as the "rebound effect", which means that although technological progress can achieve emission reductions by efficiency improvement, but it can also indirectly stimulate socio-economic growth and creates new energy demands, results in expected emission reductions being offset by the additional economic growth effect. Based on the multi-source data structure, this paper takes Yangtze-river delta urban agglomeration as an example, quantitatively evaluated the carbon rebound effect of tourism transport through the rebound effect measurement model; simulated the spatiotemporal dynamics evolution pattern of the carbon rebound effect in tourism transport through the spatial kernel density; extracted and identified the dominant factors of carbon rebound effect in tourism transport by the geographic detector. The conclusions summarized as follow: (1) The overall carbon emissions from tourism transport in the agglomeration primarily exhibit a weak rebound effect. (2) The carbon rebound effect is significantly influenced by spatiotemporal factors, which impact its development trend and interaction relations. (3) The level of tourism consumption exerts the greatest influence on the carbon rebound effect of tourism transport, while environmental regulation intensity is commonly employed as a measure to address the rebound effect. This paper aims to enhance the diversity of research on carbon emissions in tourism transport while addressing the existing limitations in spatial-temporal extension. The objective is to restrain the spread of the carbon rebound effect at the regional level, thereby providing a novel decision-making reference for the sustainable development of regional tourism.

Solvent-Free Nonthermal Destruction of PFAS Chemicals and PFAS in Sediment by Piezoelectric Ball Milling.

Studies on the destruction of solid per- and polyfluoroalkyl substances (PFAS) chemicals and PFAS-laden solid wastes significantly lag behind the urgent social demand. There is a great need to develop novel treatment processes that can destroy nonaqueous PFAS at ambient temperatures and pressures. In this study, we develop a piezoelectric-material-assisted ball milling (PZM-BM) process built on the principle that ball collisions during milling can activate PZMs to generate ∼kV potentials for PFAS destruction in the absence of solvents. Using boron nitride (BN), a typical PZM, as an example, we successfully demonstrate the complete destruction and near-quantitative (∼100%) defluorination of solid PFOS and perfluorooctanoic acid (PFOA) after a 2 h treatment. This process was also used to treat PFAS-contaminated sediment. Approximately 80% of 21 targeted PFAS were destroyed after 6 h of treatment. The reaction mechanisms were determined to be a combination of piezo-electrochemical oxidation of PFAS and fluorination of BN. The PZM-BM process demonstrates many potential advantages, as the degradation of diverse PFAS is independent of functional group and chain configurations and does not require caustic chemicals, heating, or pressurization. This pioneering study lays the groundwork for optimizing PZM-BM to treat various PFAS-laden solid wastes.

Pharmacogenetic activation of parvalbumin interneurons in the prefrontal cortex rescues cognitive deficits induced by adolescent MK801 administration.

The cognitive symptoms of schizophrenia (SZ) present a significant clinical burden. They are treatment resistant and are the primary predictor of functional outcomes. Although the neural mechanisms underlying these deficits remain unclear, pathological GABAergic signaling likely plays an essential role. Perturbations with parvalbumin (PV)-expressing fast-spiking (FS) interneurons in the prefrontal cortex (PFC) are consistently found in post-mortem studies of patients with SZ, as well as in animal models. Our studies have shown decreased prefrontal synaptic inhibition and PV immunostaining, along with working memory and cognitive flexibility deficits in the MK801 model. To test the hypothesized association between PV cell perturbations and impaired cognition in SZ, we activated prefrontal PV cells by using an excitatory DREADD viral vector with a PV promoter to rescue the cognitive deficits induced by adolescent MK801 administration in female rats. We found that targeted pharmacogenetic upregulation of prefrontal PV interneuron activity can restore E/I balance and improve cognition in the MK801 model. Our findings support the hypothesis that the reduced PV cell activity levels disrupt GABA transmission, resulting in the disinhibition of excitatory pyramidal cells. This disinhibition leads to an elevated prefrontal excitation/inhibition (E/I) balance that could be causal for cognitive impairments. Our study provides novel insights into the causal role of PV cells in cognitive function and has clinical implications for understanding the pathophysiology and management of SZ.

Three-photon excited fluorescence imaging in neuroscience: From principles to applications.

The development of three-photon microscopy (3PM) has greatly expanded the capability of imaging deep within biological tissues, enabling neuroscientists to visualize the structure and activity of neuronal populations with greater depth than two-photon imaging. In this review, we outline the history and physical principles of 3PM technology. We cover the current techniques for improving the performance of 3PM. Furthermore, we summarize the imaging applications of 3PM for various brain regions and species. Finally, we discuss the future of 3PM applications for neuroscience.

Solving the two-decades-old murder case through joint application of ZooMS and ancient DNA approaches.

Bones are one of the most common biological types of evidence in forensic cases. Discriminating human bones from irrelevant species is important for the identification of victims; however, the highly degraded bones could be undiagnostic morphologically and difficult to analyze with standard DNA profiling approaches. The same challenge also exists in archaeological studies. Here, we present an initial study of an analytical strategy that involves zooarchaeology by mass spectrometry (ZooMS) and ancient DNA methods. Through the combined strategy, we managed to identify the only biological evidence of a two-decades-old murder case - a small piece of human bone out of 19 bone fragments - and confirmed the kinship between the victim and the putative parents through joint application of next-generation sequencing (NGS) and Sanger sequencing methods. ZooMS effectively screened out the target human bone while ancient DNA methods improve the DNA yields. The combined strategy in this case outperforms the standard DNA profiling approach with shorter time, less cost, as well as higher reliability for the genetic identification results. HIGHLIGHTS: • The first application of zooarchaeology by mass spectrometry technique in the forensic case for screening out human bones from bone fragment mixtures. • Application of ancient DNA technique to recover the highly degraded DNA sequence from the challenging sample that failed standard DNA profiling approaches. • A fast, sensitive, and low-cost strategy that combines the strengths of protein analysis and DNA analysis for kinship identification in forensic research.