Zona incerta mediates early life isoflurane-induced fear memory deficits.

The potential long-term effects of anesthesia on cognitive development, especially in neonates and infants, have raised concerns. However, our understanding of its underlying mechanisms and effective treatments is still limited. In this study, we found that early exposure to isoflurane (ISO) impaired fear memory retrieval, which was reversed by dexmedetomidine (DEX) pre-treatment. Measurement of c-fos expression revealed that ISO exposure significantly increased neuronal activation in the zona incerta (ZI). Fiber photometry recording showed that ZI neurons from ISO mice displayed enhanced calcium activity during retrieval of fear memory compared to the control group, while DEX treatment reduced this enhanced calcium activity. Chemogenetic inhibition of ZI neurons effectively rescued the impairments caused by ISO exposure. These findings suggest that the ZI may play a pivotal role in mediating the cognitive effects of anesthetics, offering a potential therapeutic target for preventing anesthesia-related cognitive impairments.

Early blood immune molecular alterations in cynomolgus monkeys with a PSEN1 mutation causing familial Alzheimer's disease.

INTRODUCTION: More robust non-human primate models of Alzheimer's disease (AD) will provide new opportunities to better understand the pathogenesis and progression of AD. METHODS: We designed a CRISPR/Cas9 system to achieve precise genomic deletion of exon 9 in cynomolgus monkeys using two guide RNAs targeting the 3' and 5' intron sequences of PSEN1 exon 9. We performed biochemical, transcriptome, proteome, and biomarker analyses to characterize the cellular and molecular dysregulations of this non-human primate model. RESULTS: We observed early changes of AD-related pathological proteins (cerebrospinal fluid Aß42 and phosphorylated tau) in PSEN1 mutant (ie, PSEN1-ΔE9) monkeys. Blood transcriptome and proteome profiling revealed early changes in inflammatory and immune molecules in juvenile PSEN1-ΔE9 cynomolgus monkeys. DISCUSSION: PSEN1 mutant cynomolgus monkeys recapitulate AD-related pathological protein changes, and reveal early alterations in blood immune signaling. Thus, this model might mimic AD-associated pathogenesis and has potential utility for developing early diagnostic and therapeutic interventions. HIGHLIGHTS: A dual-guide CRISPR/Cas9 system successfully mimics AD PSEN1-ΔE9 mutation by genomic excision of exon 9. PSEN1 mutant cynomolgus monkey-derived fibroblasts exhibit disrupted PSEN1 endoproteolysis and increased Aß secretion. Blood transcriptome and proteome profiling implicate early inflammatory and immune molecular dysregulation in juvenile PSEN1 mutant cynomolgus monkeys. Cerebrospinal fluid from juvenile PSEN1 mutant monkeys recapitulates early changes of AD-related pathological proteins (increased Aß42 and phosphorylated tau).

A Novel Retrograde AAV Variant for Functional Manipulation of Cortical Projection Neurons in Mice and Monkeys.

Retrograde adeno-associated viruses (AAVs) are capable of infecting the axons of projection neurons and serve as a powerful tool for the anatomical and functional characterization of neural networks. However, few retrograde AAV capsids have been shown to offer access to cortical projection neurons across different species and enable the manipulation of neural function in non-human primates (NHPs). Here, we report the development of a novel retrograde AAV capsid, AAV-DJ8R, which efficiently labeled cortical projection neurons after local administration into the striatum of mice and macaques. In addition, intrastriatally injected AAV-DJ8R mediated opsin expression in the mouse motor cortex and induced robust behavioral alterations. Moreover, AAV-DJ8R markedly increased motor cortical neuron firing upon optogenetic light stimulation after viral delivery into the macaque putamen. These data demonstrate the usefulness of AAV-DJ8R as an efficient retrograde tracer for cortical projection neurons in rodents and NHPs and indicate its suitability for use in conducting functional interrogations.

An ultra-compact promoter drives widespread neuronal expression in mouse and monkey brains.

Promoters are essential tools for basic and translational neuroscience research. An ideal promoter should possess the shortest possible DNA sequence with cell-type selectivity. However, whether ultra-compact promoters can offer neuron-specific expression is unclear. Here, we report the development of an extremely short promoter that enables selective gene expression in neurons, but not glial cells, in the brain. The promoter sequence originates from the human CALM1 gene and is only 120 bp in size. The CALM1 promoter (pCALM1) embedded in an adeno-associated virus (AAV) genome directed broad reporter expression in excitatory and inhibitory neurons in mouse and monkey brains. Moreover, pCALM1, when inserted into an all-in-one AAV vector expressing SpCas9 and sgRNA, drives constitutive and conditional in vivo gene editing in neurons and elicits functional alterations. These data demonstrate the ability of pCALM1 to conduct restricted neuronal gene expression, illustrating the feasibility of ultra-miniature promoters for targeting brain-cell subtypes.

Circuit-specific gene therapy reverses core symptoms in a primate Parkinson's disease model.

Parkinson's disease (PD) is a debilitating neurodegenerative disorder. Its symptoms are typically treated with levodopa or dopamine receptor agonists, but its action lacks specificity due to the wide distribution of dopamine receptors in the central nervous system and periphery. Here, we report the development of a gene therapy strategy to selectively manipulate PD-affected circuitry. Targeting striatal D1 medium spiny neurons (MSNs), whose activity is chronically suppressed in PD, we engineered a therapeutic strategy comprised of a highly efficient retrograde adeno-associated virus (AAV), promoter elements with strong D1-MSN activity, and a chemogenetic effector to enable precise D1-MSN activation after systemic ligand administration. Application of this therapeutic approach rescues locomotion, tremor, and motor skill defects in both mouse and primate models of PD, supporting the feasibility of targeted circuit modulation tools for the treatment of PD in humans.

Development and characterization of an Otp conditional loss of function allele.

Orthopedia (Otp) is a homeodomain transcription factor that plays an essential role in the development of hypothalamic neurosecretory systems. Loss of Otp results in the failure of differentiation of key hypothalamic neuroendocrine cell types, and pups die soon after birth. Although the constitutive knockout Otp mouse model (Otp KO ) has significantly expanded our understanding of Otp's function in vivo, a conditional loss of function Otp allele that enables tissue or cell-type specific ablation of Otp has not been developed. Here, we used CRISPR/Cas9 gene-editing technology to generate a conditional Otp knockout mouse line in which exon 2 of the murine Otp gene is flanked by LoxP sites (Otp f/f ). Crossing the Otp f/f mouse with Agrp-Ires-cre mouse, we demonstrate the requirement for Otp in the continuous differentiation of AgRP neurons after cell fate determination. We also show that the residual AgRP neurons in Agrp-Ires-cre;Otp f/f mice project to similar downstream target regions. This newly developed Otp f/f mouse can be used to explore the functions of Otp with cell-type or temporal specificity.

Prenatal Exposure to Ketamine Leads to Anxiety-Like Behaviors and Dysfunction in Bed Nucleus of Stria Terminalis.

BACKGROUND: Both the clinical and preclinical studies have suggested embryonic or infant exposure to ketamine, a general anesthetic, pose a great threat to the developing brain. However, it remains unclear how ketamine may contribute to the brain dysfunctions. METHODS: A mouse model of prenatal exposure to ketamine was generated by i.m. injection and continuous i.p. infusion of pregnant mice. Open field test and elevated plus maze test were used to analyze the behavioral alterations induced by ketamine. Immunostaining by c-Fos was used to map the neuron activity. Chemogenetic modulation of the neurons was used to rescue the abnormal neuron activity and behaviors. RESULTS: Here we show that mice prenatally exposed to ketamine displayed anxiety-like behaviors during adulthood, but not during puberty. C-Fos immunostaining identified abnormal neuronal activity in Bed Nucleus of the Stria Terminalis, the silencing of which by chemogenetics restores the anxiety-like behaviors. CONCLUSIONS: Taken together, these results demonstrate a circuitry mechanism of ketamine-induced anxiety-like behaviors.

CT findings predict survival of patients with peripheral T cell lymphoma: a preliminary study.

Background Peripheral T-cell lymphoma (PTCL) is an uncommon disease with poor clinical outcomes. Radiological reports on the survival of patients with PTCL are scarce. The purpose of this study is to investigate the prognostic value of CT findings to predict clinical outcomes in fifty-one patients with histologically proven PTCL. Patients and methods The clinical data and CT images of all patients were retrospectively reviewed. CT features including number of involvement sites, lesion size, shape, margin, density, peritumoral invasion, intratumoral necrosis, lymph node involvement, and degree of contrast enhancement were evaluated. Univariate and multiple logistic regression analysis were used to determine the association between the clinical outcome and radiologic factors. Results Multiple site involvement, an ill-defined margin with peritumoral invasion, inhomogeneous density, and intratumoral necrosis were found to be associated with poor outcomes in univariate analysis (P < 0.05). An ill-defined margin with peritumoral invasion, was identified as an independent risk sign by further multivariate logistic regression analysis (P < 0.05). The area under the ROC curve of this CT feature was 0.745 (P < 0.05). Conclusions An ill-defined margin with peritumoral invasion was a valuable prognostic factor to predict the worse clinical outcomes in patients with PTCL.

Diagnostic Value of Autoantibodies against Ezrin in Esophageal Squamous Cell Carcinoma.

Esophageal squamous cell carcinoma (ESCC), one of the most common malignancies worldwide, is a highly aggressive and homogeneous entity occurring in esophageal squamous epithelium, and a reliable noninvasive test for early detection is needed. A recent study showed that serum autoantibodies against Ezrin could be detected in patients with pancreatic cancer. Here, we assessed whether autoantibodies against Ezrin could have diagnostic relevance for early ESCC. We analyzed autoantibodies against Ezrin in sera of 98 normal controls and 149 patients with ESCC. Ezrin autoantibodies levels were evaluated by enzyme-linked immunosorbent assay (ELISA). Results showed that higher levels of autoantibodies against Ezrin were observed in serum samples from patients with ESCC than in serum from normal controls (P < 0.0001). Based on a cutoff value of 0.319, the sensitivity and specificity of autoantibodies against Ezrin for diagnosis of ESCC were 27.5% and 95.9%, respectively. Compared with normal controls, the positive rate of autoantibodies against Ezrin was significantly elevated in patients with early-stage ESCC (P < 0.0001). Moreover, there was no significant difference of positivity of autoantibodies against Ezrin in ESCC patients categorized according to age, gender, tumor size, tumor invasion depth, tumor site, histological grade, lymph node status, or tumor stage. Our study indicates that the presence of autoantibodies against Ezrin is significantly associated with ESCC.

Radiological-pathological correlation of yolk sac tumor in 20 patients.

BACKGROUND: Yolk sac tumor (YST) is a rare tumor. Familiarity of its radiological characteristics may permit preoperative diagnosis and improve surgical management of patients. However, a detailed description of the imaging features of YST with pathological correlation in particular is scarce. PURPOSE: To investigate computed tomography (CT) findings of YSTs with pathological correlation. MATERIAL AND METHODS: CT images of 20 patients with pathologically proven YST were retrospectively reviewed. The location, size, margin, internal architecture, and pattern and degree enhancement of the lesion were evaluated. Radiological findings were correlated with pathological results. RESULTS: The locations of 20 tumors were distributed between the testis (n = 3), ovary (n = 6), sacrococcygeal area (n = 6), rectum (n = 1), and mediastinum (n = 4). The median age was 13 years. On CT images, all tumors were seen as oval (n = 14) or irregular (n = 6), well-defined (n = 16) or ill-defined (n = 4) masses with a mean size of 9.7 cm. The lesions were solid cystic (n = 10), entirely solid (n = 6), or predominantly cystic (n = 4). Intratumoral hemorrhage, calcification, and fatty tissue were seen in nine, three, and two tumors, respectively. Discontinuity of the tumor wall was seen in eight tumors. After contrast media administration, most tumors showed heterogeneous moderate to marked enhancement (n = 7) or heterogeneous marked enhancement (n = 9). Enlarged intratumoral vessels were seen in 17 tumors. CONCLUSION: YST usually appears as a large solid-cystic mass with intratumoral hemorrhage, capsular tear, marked heterogeneous enhancement, and enlarged intratumoral vessels on CT images. Intratumoral calcification and fatty tissue, although rare, may indicate a mixed YST containing teratoma component.

CT imaging of ovarian yolk sac tumor with emphasis on differential diagnosis.

Ovarian yolk sac tumors (YSTs) are rare neoplasms. No radiological study has been done to compare the imaging findings between this type of tumor and other ovarian tumors. Here we analyzed the CT findings of 11 pathologically proven ovarian YSTs and compared their imaging findings with 18 other types of ovarian tumors in the same age range. Patient age, tumor size, tumor shape, ascites and metastasis of two groups did not differ significantly (P > 0.05). A mixed solid-cystic nature, intratumoral hemorrhage, marked enhancement and dilated intratumoral vessel of two groups differed significantly (P < 0.05). The area under the ROC curve of four significant CT features was 0.679, 0.707, 0.705, and 1.000, respectively. Multivariate logistic regression analysis identified two independent signs of YST: intratumoral hemorrhage and marked enhancement. Our results show that certain suggestive CT signs that may be valuable for improving the accuracy of imaging diagnosis of YST and may be helpful in distinguishing YST from other ovarian tumors.

Determining the organ of origin of large pelvic masses in females using multidetector CT angiography and three-dimensional volume rendering CT angiography.

OBJECTIVE: To study the value of tumour feeding arteries and the ovarian vein in determining the organ of origin of large pelvic tumours in females using multidetector CT. METHODS: One hundred and thirty patients with 131 pathologically proven tumours (>6.5 cm) were retrospectively reviewed. Conventional CT images and CT angiography were evaluated, with focus on assessing the value of tumour feeding arteries and the ovarian vein in differentiating ovarian from non-ovarian tumours. RESULTS: For 97 ovarian tumours, the feeding arteries included the ovarian artery (n = 51) and the ovarian branch of uterine artery (n = 64). For 34 non-ovarian tumours, the feeding arteries included the ovarian artery (n = 2), the uterine artery (n = 21), the mesenteric artery (n = 5), and the internal iliac artery (n = 1). The ovarian vein was identified in 86 ovarian tumours and 12 non-ovarian tumours. When the feeding arteries and the ovarian vein were combined to confirm ovarian origin, the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 97.8 %, 84.2 %, 93.8 %, 94.1 %, and 93.9 %, respectively. The accuracy was significantly higher than that of independently using the ovarian vein or the ovarian feeding arteries. CONCLUSION: Combined application of tumour feeding arteries and the ovarian vein is valuable to differentiate large ovarian from non-ovarian tumours. KEY POINTS: • CT is a valuable modality for diagnosing pelvic tumours. • Determining the organ of origin is difficult for large pelvic tumours. • Contrast-enhanced CT and CT angiography are helpful in depicting abdominopelvic vessels. • Tracking tumour-associated vessels can help differentiate large ovarian from non-ovarian tumours.

Evaluation of the relationship between extremity soft tissue sarcomas and adjacent major vessels using contrast-enhanced multidetector CT and three-dimensional volume-rendered CT angiography: a preliminary study.

BACKGROUND: Accurate description of the relationship between extremity soft tissue sarcoma and the adjacent major vessels is crucial for successful surgery. In addition to magnetic resonance imaging (MRI) or in patients who cannot undergo MRI, two-dimensional (2D) postcontrast computed tomography (CT) images and three-dimensional (3D) volume-rendered CT angiography may be valuable alternative imaging techniques for preoperative evaluation of extremity sarcomas. PURPOSE: To preoperatively assess extremity sarcomas using multidetector CT (MDCT), with emphasis on postcontrast MDCT images and 3D volume-rendered MDCT angiography in evaluating the relationship between tumors and adjacent major vessels. MATERIAL AND METHODS: MDCT examinations were performed on 13 patients with non-metastatic extremity sarcomas. Conventional CT images and 3D volume-rendered CT angiography were evaluated, with focus on the relationship between tumors and adjacent major vessels. Kappa consistency statistics were performed with surgery serving as the reference standard. RESULTS: The relationship between sarcomas and adjacent vessels was described as one of three patterns: proximity, adhesion, and encasement. Proximity was seen in five cases on postcontrast CT images or in eight cases on volume-rendered images. Adhesion was seen in three cases on both postcontrast CT images and volume-rendered images. Encasement was seen in five cases on postcontrast CT images or in two cases on volume-rendered images. Compared to surgical results, postcontrast CT images had 100% sensitivity, 83.3% specificity, 87.5% positive predictive value, 100% negative predictive value, and 92.3% accuracy in the detection of vascular invasion (κ = 0.843, P = 0.002). 3D volume-rendered CT angiography had 71.4% sensitivity, 100% specificity, 100% positive predictive value, 75% negative predictive value, and 84.6% accuracy in the detection of vascular invasion (κ = 0.698, P = 0.008). On volume-rendered images, all cases with adhesion or encasement had arterial stenosis and all tumors' feeding arteries were clearly depicted. CONCLUSION: 2D postcontrast CT images are superior to 3D volume-rendered CT angiography in evaluating the relationship between extremity sarcomas and adjacent major vessels. 3D volume-rendered CT angiography is good at assessing the tumor's blood supply, the longitudinal extent of vascular involvement, and the vascular narrowing due to the tumor.