INTRODUCTION:Dementia with Lewy bodies (DLB), a prevalent neurodegenerative dementia, involves α-synuclein (α-syn) aggregates and frequent amyloid beta (Aβ) co-pathology, but mechanistic drivers remain unclear. METHODS:We crossed pink1 knockout with APP/PS1 mice, and assessed behavioral and pathological phenotypes of the resulting animals. We also performed biochemical and biophysical characterizations of PTEN-induced kinase 1 (PINK1) phosphorylation of α-syn. RESULTS:DLB brains show PINK1 deficiency alongside α-syn and Aβ co-pathology. Mirroring human DLB patients, APP/PS1::pink1-/- mice spontaneously develop Lewy pathology at endogenous α-syn levels, affecting both central and peripheral nervous systems with heterogeneous phenotypes. Mechanistically, PINK1 phosphorylates α-syn at Thr44, suppressing Aβ-induced α-syn aggregation. Moreover, pT44-α-syn levels are correlated with PINK1 expression and activity in human brains. DISCUSSION:PINK1 deficiency synergizes with Aβ to promote Lewy pathology via loss of protective α-syn phosphorylation. The APP/PS1::pink1-/- model recapitulates key DLB features without α-syn overexpression, offering a valuable tool for future mechanistic and therapeutic studies. HIGHLIGHTS:PTEN-induced kinase 1 (PINK1) deficiency, either through reduced expression or impaired activity, is found in human dementia with Lewy bodies (DLB) patients with amyloid beta (Aβ) co-pathology. PINK1 specifically phosphorylates α-synuclein at Thr44, inhibiting Aβ-induced aggregation and preventing the development of Lewy pathology. The APP/PS1::pink1-/- mouse model recapitulates key features of human DLB, exhibiting widespread Lewy pathology and heterogeneous phenotypes. PINK1 alterations emerge as a novel genetic risk factor for DLB, opening new avenues for diagnosis and therapeutic intervention.

Microglia, as resident macrophages in the central nervous system (CNS), have been the focus of the scientific community. The pace of exploration in the origin and development of microglia, though tortuous, never stops. Since colony-stimulating factor receptor 1 (CSF1R) inhibitors can achieve effective depletion of microglia and the repopulated microglia can be comparable to the controls, the therapeutic potential of this repopulation has prompted increasing attention and investigation. Meanwhile, single-cell sequencing technology (scRNA-seq) revealed cell fate determination and cell heterogeneity of microglia during development, homeostasis, and pathological states, identifying various functional subpopulations, including disease-associated microglia (DAM), neurodegenerative microglia (MGnD), and others. Thus, novel therapeutic values are bestowed on the repopulated microglia given their strong self-renewal capacity and highly proliferative and migratory abilities. In this paper, we first provide a comprehensive summary of the exploration process concerning the origin, development, repopulation, and heterogeneity of microglia. Moreover, we emphasize the implications of microglial repopulation in CNS disorders in the era of single-cell to enhance the understanding of microglial repopulation and provide more insights for new therapeutic strategies.

Overgeneralization of fear to harmless situations is a core feature of anxiety disorders resulting from acute stress, yet the mechanisms by which fear becomes generalized are poorly understood. In this study, we show that generalized fear in mice results from a transmitter switch from glutamate to γ-aminobutyric acid (GABA) in serotonergic neurons of the lateral wings of the dorsal raphe. Similar change in transmitter identity was found in the postmortem brains of individuals with posttraumatic stress disorder (PTSD). Overriding the transmitter switch in mice prevented the acquisition of generalized fear. Corticosterone release and activation of glucocorticoid receptors mediated the switch, and prompt antidepressant treatment blocked the cotransmitter switch and generalized fear. Our results provide important insight into the mechanisms involved in fear generalization.

Nicotinamide phosphoribosyltransferase (NAMPT) is the key enzyme in the salvaging synthesize pathway of nicotinamide adenine dinucleotide (NAD). The neuroprotective roles of NAMPT on neurodegeneration have been widely explored in aging brain and Alzheimer’s Disease. However, its roles in Parkinson’s Disease (PD) remain to be elucidated. We found that the dopaminergic neurons in substantia nigra expressed higher levels of NAMPT than the other types of neurons. Using conditional knockout of the nampt gene in dopaminergic neurons and utilizing a NAMPT inhibitor in the substantia nigra of mice, we found that the NAMPT deficiency triggered the loss of dopaminergic neurons, the impairment of the dopamine nigrostriatal pathway, and the development of PD-like motor dysfunction. In the rotenone-induced PD mouse model, nicotinamide ribose (NR), a precursor of NAD, rescued the loss of dopaminergic neurons, the impairment of dopamine nigrostriatal pathway, and mitigated PD-like motor dysfunction. In SH-SY5Y cells, NAD suppression induced the accumulation of reactive oxygen species (ROS), mitochondrial impairment, and cell death, which was reversed by N-acetyl cysteine, an antioxidant and ROS scavenger. Rotenone decreased NAD level, induced the accumulation of ROS and the impairment of mitochondria, which was reversed by NR. In summary, our findings show that NAMPT deficiency is an independent risk factor for the development of PD, and NAD precursors have the potential to rescue degenerative dopaminergic neurons and treat PD.

Poly (ADP-ribose) polymerase-1 (PARP-1) activity significantly increases during cerebral ischemia/reperfusion. PARP-1 is an NAD+-consumption enzyme. PARP-1 hyperactivity causes intracellular NAD+ deficiency and bioenergetic collapse, contributing to neuronal death. Besides, the powerful trigger of PARP-1 causes the catalyzation of poly (ADP-ribosyl)ation (PARylation), a posttranslational modification of proteins. Here, we found that PARP-1 was activated in the ischemic brain tissue during middle-cerebral-artery occlusion and reperfusion (MCAO/R) for 24 h, and PAR accumulated in the neurons in mice. Using immunoprecipitation, Western blotting, liquid chromatography-mass spectrometry, and 3D-modeling analysis, we revealed that the activation of PARP-1 caused PARylation of hexokinase-1 and lactate dehydrogenase-B, which, therefore, caused the inhibition of these enzyme activities and the resulting cell energy metabolism collapse. PARP-1 inhibition significantly reversed the activity of hexokinase and lactate dehydrogenase, decreased infarct volume, and improved neuronal deficiency. PARP-1 inhibitor combined with pyruvate further alleviated MCAO/R-induced ischemic brain injury in mice. As such, we conclude that PARP-1 inhibitor alleviates neuronal death partly by inhibiting the PARylation of metabolic-related enzymes and reversing metabolism reprogramming during cerebral ischemia/reperfusion injury in mice. PARP-1 inhibitor combined with pyruvate might be a promising therapeutic approach against brain ischemia/reperfusion injury.

Nicotinamide phosphoribosyltransferase (NAMPT) is the key enzyme in the salvaging synthesize pathway of nicotinamide adenine dinucleotide (NAD). The neuroprotective roles of NAMPT on neurodegeneration have been explored in aging brain and Alzheimer’s Disease. However, its roles in Parkinson’s Disease (PD) remain to be elucidated. We found that the dopaminergic neurons in substantia nigra expressed higher levels of NAMPT than the other types of neurons. Using conditional knockout of the Nampt gene in dopaminergic neurons and utilizing a NAMPT inhibitor in the substantia nigra of mice, we found that the NAMPT deficiency triggered the time-dependent loss of dopaminergic neurons, the impairment of the dopamine nigrostriatal pathway, and the development of PD-like motor dysfunction. In the rotenone-induced PD mouse model, nicotinamide ribose (NR), a precursor of NAD, rescued the loss of dopaminergic neurons, the impairment of dopamine nigrostriatal pathway, and mitigated PD-like motor dysfunction. In SH-SY5Y cells, NAD suppression induced the accumulation of reactive oxygen species (ROS), mitochondrial impairment, and cell death, which was reversed by N-acetyl cysteine, an antioxidant and ROS scavenger. Rotenone decreased NAD level, induced the accumulation of ROS and the impairment of mitochondria, which was reversed by NR. In summary, our findings show that the ablation of NAMPT in dopaminergic neurons leads to neurodegeneration and contributes to the development of PD. The NAD precursors have the potential to protect the degeneration of dopaminergic neurons, and offering a therapeutic approach for the treatment of PD.

PDF file - 169K, Supplementary Table S1. The clinical features of the glioma specimens used in this study. Supplementary Table S2. Primers used for qRT-PCT, miRNA cloning and luciferase plasmid construction. Supplementary Table S3. Top 20 downregulated miRNAs in tumor tissue (GBM-T) versus adjacent normal tissue (GBM-N). Supplementary Table S4. Top 20 upregulated miRNAs in tumor tissue (GBM-T) versus adjacent normal tissue (GBM-N). Supplementary Table S5. The correlation between miR-663 expression and clinicopathological factors of patients with astrocytoma (Grade I and II). Supplementary Table S6. The correlation between miR-663 expression and clinicopathological factors of patients with anaplastic astrocytoma (Grade III). Supplementary Table S7. The correlation between miR-663 expression and clinicopathological factors of patients with GBM (Grade IV). Supplementary Table S8. Univariate analysis for disease-free survival and overall survival in GBM patients. Supplementary Table S9. Top 30 overlapped predicted targets of miR-663 and their degrees based on signal transduction network.

Accurate pathologic diagnosis and molecular classification of breast mass biopsy tissue is important for determining individualized therapy for (neo)adjuvant systemic therapies for invasive breast cancer. The CassiII rotational core biopsy system is a novel biopsy technique with a guide needle and a “stick-freeze” technology. The comprehensive assessments including the concordance rates of diagnosis and biomarker status between CassiII and core needle biopsy were evaluated in this study. Estrogen receptor (ER), progesterone receptor (PgR), human epidermal growth factor receptor 2 (HER2), and Ki67 were analyzed through immunohistochemistry. In total, 655 patients with breast cancer who underwent surgery after biopsy at Sir Run Run Shaw Hospital between January 2019 to December 2021 were evaluated. The concordance rates (CRs) of malignant surgical specimens with CassiII needle biopsy was significantly high compared with core needle biopsy. Moreover, CassiII needle biopsy had about 20% improvement in sensitivity and about 5% improvement in positive predictive value compared to Core needle biopsy. The characteristics including age and tumor size were identified the risk factors for pathological inconsistencies with core needle biopsies. However, CassiII needle biopsy was associated with tumor diameter only. The CRs of ER, PgR, HER2, and Ki67 using Cassi needle were 98.08% (kappa, 0.941; p<.001), 90.77% (kappa, 0.812; p<.001), 69.62% (kappa, 0.482; p<.001), and 86.92% (kappa, 0.552; p<.001), respectively. Post-biopsy complications with CassiII needle biopsy were also collected. The complications of CassiII needle biopsy including chest stuffiness, pain and subcutaneous ecchymosis are not rare. The underlying mechanism of subcutaneous congestion or hematoma after CassiII needle biopsy might be the larger needle diameter and the effect of temperature on coagulation function. In summary, CassiII needle biopsy is age-independent and has a better accuracy than CNB for distinguishing carcinoma in situ and invasive carcinoma.

Abstract Purpose Vacuum-assisted breast biopsy (VABB) offers high accuracy in the diagnosis of breast tumors but makes the location of the original lesion difficult to identify when the subsequent breast-conserving surgery (BCS) is to be carried out. The aim of this study was to describe an effective technique to identify the exact location of the VABB cavity and to make breast-conserving resection more precise. Methods In this study, a balloon urinary catheter was inserted through the biopsy track; thus, the inflated balloon and catheter perfectly reconstructed the VABB cavity and biopsy track. Then breast-conserving resection with the guidance of the catheter was performed. Six patients were enrolled and assessed for their surgical findings, pathological results, surgical margins, and postoperative complications. Results Residual tumors were confirmed in 4 out of 6 patients after VABB. All the patients gained negative margins and underwent successful breast-conserving surgery. None of them required re-excisions. Satisfactory cosmetic results were observed after the catheter-directed BCS. Conclusions The results of this study show the effectiveness of urinary catheter-guided breast-conserving surgery after VABB. The procedure is simple and inexpensive. This new technique enabled precise breast-conserving resection of the targeted lesion and potentially improve successful rate of breast-conserving surgery.

Overgeneralization of fear to harmless situations is a core feature of anxiety disorders resulting from acute stress, yet the mechanisms by which fear becomes generalized are poorly understood. Here we show that generalized fear in mice in response to footshock results from a transmitter switch from glutamate to GABA in serotonergic neurons of the lateral wings of the dorsal raphe. We observe a similar change in transmitter identity in the postmortem brains of PTSD patients. Overriding the transmitter switch in mice using viral tools prevents the acquisition of generalized fear. Corticosterone release and activation of glucocorticoid receptors trigger the switch, and prompt antidepressant treatment blocks the co-transmitter switch and generalized fear. Our results provide new understanding of the plasticity involved in fear generalization.

Supplementary Figure S4. The prognostic significance of miR-663 and CXCR4 in astrocytic glioma.

Supplementary Figure S1. Luciferase reporter assay of U87-MG cell transfected with miR-663 mimics or control oligomer, together with CXCR4-CDS wild-type reporter (CXCR4-CDS-wt) or the indicated mutant ones.

PDF file - 1654K, Supplementary Fig. S4. The suppressive effects of miR-663 on PIK3CD and its downstream proliferation and invasion-related targets.

Abstract Purpose Vacuum-assisted breast biopsy (VABB) offers high accuracy in the diagnosis of breast tumors but makes the location of the original lesion difficult to identify when the subsequent breast-conserving surgery (BCS) is to be carried out. The aim of this study was to describe an effective technique to identify the exact location of the VABB cavity and to make breast-conserving resection more precise. Methods In this study, a balloon urinary catheter was inserted through the biopsy track; thus, the inflated balloon and catheter perfectly reconstructed the VABB cavity and biopsy track. Then breast-conserving resection with the guidance of the catheter was performed. Six patients were enrolled and assessed for their surgical findings, pathological results, surgical margins, and postoperative complications. Results Residual tumors were confirmed in 4 out of 6 patients after VABB. All the patients gained negative margins and underwent successful breast-conserving surgery. None of them required re-excisions. Satisfactory cosmetic results were observed after the catheter-directed BCS. Conclusions The results of this study show the effectiveness of urinary catheter-guided breast-conserving surgery after VABB. The procedure is simple and inexpensive. This new technique enabled precise breast-conserving resection of the targeted lesion and potentially improve successful rate of breast-conserving surgery.

Supplementary Figure S3. Correlation between miR-663 and CXCR4 mRNA in grade II and III astrocytic glioma specimens.

Supplementary Figure legends. Supplementary Figure legends for Fig. S1-S4.

Ser65-phosphorylated ubiquitin (pUb) was found elevated in neurons of aged and neurodegenerative brains. Yet little is known whether a causative link exists between pUb level and brain aging. Here we show that the knockout of pink1 , a Ub kinase, abolished pUb elevation and decelerated protein aggregation in aged mouse brains and cells with proteasomal inhibition. Conversely, over-expression of PINK1 but not the kinase-dead version increased the pUb level and accelerated protein aggregation by suppressing of proteasomal degradation. Furthermore, PINK1 over-expression in mouse hippocampus neurons increased pUb level and protein aggregation, slowly leading to mitochondrial injury, neurodegeneration, and cognitive impairment. Notably, the neuronal damages induced by PINK1 were rescued by the dominant negative Ub/S65A mutant, while Ub/S65E phosphomimetic mutant caused neuronal death. Together, an incidental increase of Ub phosphorylation can progressively and cumulatively cause the decline of Ub-dependent proteasomal activity, consequenting promotes neurodegeneration in the aging brain.

The tumor microenvironment is proposed to contribute substantially to the progression of cancers, including breast cancer. Cancer-associated fibroblasts (CAFs) are the most abundant components of the tumor microenvironment. Studies have revealed that CAFs in breast cancer originate from several types of cells and promote breast cancer malignancy by secreting factors, generating exosomes, releasing nutrients, reshaping the extracellular matrix, and suppressing the function of immune cells. CAFs are also becoming therapeutic targets for breast cancer due to their specific distribution in tumors and their unique biomarkers. Agents interrupting the effect of CAFs on surrounding cells have been developed and applied in clinical trials. Here, we reviewed studies examining the heterogeneity of CAFs in breast cancer and expression patterns of CAF markers in different subtypes of breast cancer. We hope that summarizing CAF-related studies from a historical perspective will help to accelerate the development of CAF-targeted therapeutic strategies for breast cancer.