A novel heterozygous variant of the SALL1 gene with atypical Townes-Brocks syndrome phenotypes in Chinese family.

Townes-Brocks syndrome (TBS) is an autosomal dominant disorder characterised by the triad of anorectal, thumb, and ear malformations. It may also be accompanied by defects in kidney, heart, eyes, hearing, and feet. TBS has been demonstrated to result from heterozygous variants in the SALL1 gene, which encodes zinc finger protein believed to function as a transcriptional repressor. The clinical characteristics of an atypical TBS phenotype patient from a Chinese family are described, with predominant manifestations including external ear dysplasia, unilateral renal hypoplasia with mild renal dysfunction, and hearing impairment. A novel heterozygous variant c.3060T>A (p.Tyr1020*) in exon 2 of the SALL1 gene was identified in this proband. Pyrosequencing of the complementary DNA of the proband revealed that the variant transcript accounted for 48% of the total transcripts in peripheral leukocytes, indicating that this variant transcript has not undergone nonsense-mediated mRNA decay. This variant c.3060T > A is located at the terminal end of exon 2, proximal to the 3' end of the SALL1 gene, and exerts a relatively minor impact on protein function. We suggest that the atypical TBS phenotype observed in the proband may be attributed to the truncated protein retaining partial SALL1 function.

Valosin-containing protein acts as a target and mediator of S-nitrosylation in the heart through distinct mechanisms.

S-nitrosylation (SNO) is an emerging paradigm of redox signaling protecting cells against oxidative stress in the heart. Our previous studies demonstrated that valosin-containing protein (VCP), an ATPase-associated protein, is a vital mediator protecting the heart against cardiac stress and ischemic injury. However, the molecular regulations conferred by VCP in the heart are not fully understood. In this study, we explored the potential role of VCP in cardiac protein SNO using multiple cardiac-specific genetically modified mouse models and various analytical techniques including biotin switch assay, liquid chromatography, mass spectrometry, and western blotting. Our results showed that cardiac-specific overexpression of VCP led to an overall increase in the levels of SNO-modified cardiac proteins in the transgenic (TG) vs. wild-type (WT) mice. Mass spectrometry analysis identified mitochondrial proteins involved in respiration, metabolism, and detoxification as primary targets of SNO modification in VCP-overexpressing mouse hearts. Particularly, we found that VCP itself underwent SNO modification at a specific cysteine residue in its N-domain. Additionally, our study demonstrated that glyceraldehyde 3-phosphate dehydrogenase (GAPDH), a key enzyme in glycolysis, also experienced increased SNO in response to VCP overexpression. While deletion of inducible nitric oxide synthase (iNOS) in VCP TG mice did not affect VCP SNO, it did abolish SNO modification in mitochondrial complex proteins, suggesting a dual mechanism of regulation involving both iNOS-dependent and independent pathways. Overall, our findings shed light on post-translational modification of VCP in the heart, unveiling a previously unrecognized role for VCP in regulating cardiac protein SNO and offering new insights into its function in cardiac protection.

Impact of Postoperative Radiotherapy on the Prognosis of Early-Stage (pT1-2N0M0) Oral Tongue Squamous Cell Carcinoma.

PURPOSE: To identify subgroups of patients with early-stage (pT1-2N0M0) oral tongue squamous cell carcinoma (OTSCC) who may benefit from postoperative radiotherapy (PORT). PATIENTS AND METHODS: This retrospective cohort study included 528 patients diagnosed between October 2009 and December 2021. Clinicopathological characteristics and treatments with or without PORT were analyzed for their impact on outcomes. RESULTS: Among 528 patients who underwent radical surgery (median age, 62 years [IQR, 52-69]), 145 (27.5%) also underwent PORT. Multivariate analyses revealed that PORT was associated with improved survival outcomes, whereas moderate-to-poor differentiation, perineural infiltration (PNI), lymphovascular invasion (LVI), and increasing depth of invasion (DOI) were associated with poorer survival outcomes. For patients with moderate-to-poor differentiation, the surgery + PORT group showed improved outcomes compared with the surgery-alone group. After propensity score matching, the results were as follows: overall survival (OS), 97% versus 69%, P = .003; disease-free survival (DFS), 88% versus 50%, P = .001. After excluding cases with PNI/LVI, the differences persisted: OS, 97% versus 82%, P = .040; DFS, 87% versus 64%, P = .012. Similar survival benefits were observed in 104 patients with PNI and/or LVI (OS, 81% v 58%; P = .022; DFS, 76% v 47%; P = .002). In subgroups with DOI >5 mm or close margins, PORT contributed to improved DFS (80% v 64%; P = .006; 92% v 66%; P = .049) but did not significantly affect OS. CONCLUSION: Patients with moderately-to-poorly differentiated pT1-2N0M0 OTSCC benefited from PORT. Our study provided evidence that patients with PNI and/or LVI who underwent PORT had improved survival. PORT also offered DFS benefit among patients with DOI >5 mm.

Analyzing the Correlation Between Serum Biomarkers and Child-Pugh Score in Liver Cirrhosis.

Objective: This study aimed to analyze the diagnostic efficacy of serum biomarkers in liver cirrhosis patients categorized by Child-Pugh scores. Methods: An observational cross-sectional study design was employed. A total of 110 liver cirrhosis patients, classified according to Child-Pugh scores and 60 healthy individuals were included in this study. Serum levels of adenosine deaminase (ADA), adiponectin (APN), matrix metalloproteinase-2 (MMP-2), alkaline phosphatase (ALP), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were measured. Results: The levels of ADA, APN, MMP-2, ALP, ALT, and AST were significantly higher in the study group compared to the control group (P < .05). Furthermore, these levels increased with the severity of liver cirrhosis, with higher levels observed in patients with Child-Pugh class C. The positive diagnostic rates for joint detection in Child-Pugh class A, B, and C were 93.75% (30/32), 100% (34/34), and 100% (44/44), respectively. Conclusions: Combined detection of serum biomarkers improves the diagnostic efficacy of liver cirrhosis. The diagnostic rates were higher when considering Child-Pugh scores, with the highest rates observed in class C.

Functional analysis of the CTNS gene exonic variants predicted to affect splicing.

Cystinosis is a severe, monogenic systemic disease caused by variants in CTNS gene. Currently, there is growing evidence that exonic variants in many diseases can affect pre-mRNA splicing. The impact of CTNS gene exonic variants on splicing regulation may be underestimated due to the lack of routine studies at the RNA level. Here, we analyzed 59 exonic variants in the CTNS gene using bioinformatics tools and identified candidate variants that may induce splicing alterations by minigene assays. We identified six exonic variants that induce splicing alterations by disrupting the ratio of exonic splicing enhancers/exonic splicing silencers (ESEs/ESSs) or by interfering with the recognition of classical splice sites, or both. Our results help in the correct molecular characterization of variants in cystinosis and inform emerging therapies. Furthermore, our work suggests that the combination of in silico and in vitro assays facilitates to assess the effects of DNA variants driving rare genetic diseases on splicing regulation and will enhance the clinical utility of variant functional annotation.

Adult localized Langerhans cell histiocytosis: A case report.

BACKGROUND: Langerhans cell histiocytosis (LCH) is a rare clonal proliferative disease of Langerhans cells with unknown pathogenesis. An increasing number of clinicians recognize that LCH has a wide clinical spectrum and a highly varied course. Adults rarely develop LCH. Here, we report a case of adult localized LCH. CASE SUMMARY: A 32-year-old woman presented with plaques and ulcers on the vulva and crissum, accompanied by pain that persisted for more than one year. Physical examination revealed a red-infiltrating plaque with ulcerations and exudates in the vulva and crissum. Pathological examination revealed a diffuse infiltration of lymphocytes, eosinophilic granulocytes, and histiocytoid cells in the superficial dermis. Proliferative histiocytoid cells showed mild atypia, partly with kidney-shaped nuclei. Immunohistochemical examination showed that the histiocytoid cells were positive for S100 protein and CD1 and weakly positive for CD68 (20% +), with a Ki-67 index of 30%. Laboratory tests did not reveal any other systemic damage. The patient was diagnosed with adult localized LCH and was prescribed oral prednisone (20 mg) once daily. The skin lesions gradually improved and are still being followed-up. CONCLUSION: Adult localized LCH is rare and must be differentiated from other common conditions.

Nuclear translocation of thioredoxin-1 promotes colorectal cancer development via modulation of the IL-6/STAT3 signaling axis through interaction with STAT3.

Background: Thioredoxin 1 (Trx-1) is a small redox protein predominantly localized in the cytoplasm. Its expression is increased in several cancers, including colorectal cancer (CRC). However, the function of Trx-1 translocation to the nucleus in cancer is not clear. In this study, we investigated the role of Trx-1 nuclear translocation in development of CRC. Methods: Expression of Trx-1 and STAT3 was analyzed by Western blot and immunofluorescence. Endogenous interaction of Trx-1, STAT3, and karyopherin α1 in CRC cells was analyzed by co-immunoprecipitation. Trx-1 and pSTAT3 nuclear staining in human CRC tissues was analyzed by immunohistochemistry. A mouse model of AOM/DSS induced colitis-associated cancer (CAC) was utilized to investigate the antitumor effect of PX-12, a Trx-1 inhibitor. A knockin mouse with the Txn1(KK81-82EE) mutation was generated via CRISPR/Cas9, and CAC was induced in knockin and wild-type mice. Results: Nuclear translocation of Trx-1 was induced by IL-6, and inhibition of this translocation reversed IL-6-induced epithelial-to-mesenchymal transition, invasion and metastasis. Karyopherin α1 was found to specifically mediate IL-6-induced translocation of the Trx-1-pSTAT3 complex into the nucleus. Nuclear Trx-1 expression was closely correlated with lymph node metastasis and distant metastasis in human CRC. In addition, nuclear staining of Trx-1 showed significant positive correlation with nuclear staining of pSTAT3 in human CRC tissues. PX-12, an inhibitor of Trx-1, significantly impaired the activation of STAT3 and suppressed the development of AOM/DSS-induced CAC in mice. Moreover, AOM/DSS-induced nuclear Trx-1 expression was suppressed in Txn1(KK81-82EE) mice, which inhibited STAT3 activation and cancer progression. Conclusions: These results provide new insights into the mechanisms of STAT3 activation triggered by IL-6 and identify nuclear translocation of Trx-1 as a potential therapeutic target for the treatment of CRC and CAC.

Application of carbon nanoparticles combined with refined extracapsular anatomy in endoscopic thyroidectomy.

Objective: To evaluate the value of refined extracapsular anatomy combined with carbon nanoparticle suspension tracing technology for protecting parathyroid function and the thoroughness of lymph node dissection in the central region during endoscopic thyroid cancer surgery. Patients and methods: Retrospective clinical data analysis was performed on 108 patients who underwent endoscopic thyroid cancer surgery at the First Affiliated Hospital of Wannan Medical College (Yijishan Hospital) from November 2019 to November 2022. Before surgery, thyroid function tests, color Doppler ultrasounds and neck-enhanced CT scans were performed on all patients. Cytopathological diagnosis obtained via ultrasound-guided fine-needle aspiration served as confirmation for the primary diagnosis. It was determined whether to perform a total thyroidectomy or a hemithyroidectomy (HT) together with preventive unilateral (ipsilateral) central neck dissection. Follow-up times were 1 to 34 months. Results: Transient neuromuscular symptoms were present in 3.70% (4/108) cases, with no permanent neuromuscular symptoms or permanent hypoparathyroidism. Regarding transient hypoparathyroidism, the patients recovered after three months and did not need long-term calcium supplementation. The number of harvested LNs (mean± SD) was 5.54 ± 3.84, with ≤5 in 57.41% (62/108) and >5 in 42.59% (46/108) cases. The number of patients with metastatic LNs was 37.96% (41/108), with ≤2 in 65.85% (27/41) and >2 in 34.15% (14/41) cases. Conclusions: Fine extracapsular anatomy combined with carbon nanoparticle suspension tracing is effective in endoscopic thyroid cancer surgery. It can improve the thoroughness of prophylactic central neck dissection and recognition of the parathyroid gland and avoid parathyroid injury and other complications to effectively protect parathyroid function.

Image quality of coronary CT angiography at ultra low tube voltage reconstructed with a deep-learning image reconstruction algorithm in patients of different weight.

Background: GE Healthcare's new generation of deep-learning image reconstruction (DLIR), the Revolution Apex CT is the first CT image reconstruction engine based on a deep neural network to be approved by the US Food and Drug Administration (FDA). It can generate high-quality CT images that restore the true texture with a low radiation dose. The aim of the present study was to assess the image quality of coronary CT angiography (CCTA) at 70 kVp with the DLIR algorithm as compared to the adaptive statistical iterative reconstruction-Veo (ASiR-V) algorithm in patients of different weight. Methods: The study group comprised 96 patients who underwent CCTA examination at 70 kVp and were subdivided by body mass index (BMI) into normal-weight patients [48] and overweight patients [48]. ASiR-V40%, ASiR-V80%, DLIR-low, DLIR-medium, and DLIR-high images were obtained. The objective image quality, radiation dose, and subjective score of the two groups of images with different reconstruction algorithms were compared and statistically analyzed. Results: In the overweight group, the noise of the DLIR image was lower than that of the routinely used ASiR-40%, and the contrast-to-noise ratio (CNR) of DLIR (H: 19.15±4.31; M: 12.68±2.91; L: 10.59±2.32) was higher than that of the ASiR-40% reconstructed image (8.39±1.46), with statistically significant differences (all P values <0.05). The subjective image quality evaluation of DLIR was significantly higher than that of ASiR-V reconstructed images (all P values <0.05), with the DLIR-H being the best. In a comparison of the normal-weight and overweight groups, the objective score of the ASiR-V-reconstructed image increased with increasing strength, but the subjective image evaluation decreased, and both differences (i.e., objective and subjective) were statistically significant (P<0.05). In general, the objective score of the DLIR reconstruction image between the two groups increased with increased noise reduction, and the DLIR-L image was the best. The difference between the two groups was statistically significant (P<0.05), but there was no significant difference in subjective image evaluation between the two groups. The effective dose (ED) of the normal-weight group and the overweight group was 1.36±0.42 and 1.59±0.46 mSv, respectively, and was significantly higher in the overweight group (P<0.05). Conclusions: As the strength of the ASiR-V reconstruction algorithm increased, the objective image quality increased accordingly, but the high-strength ASiR-V changed the noise texture of the image, resulting in a decrease in the subjective score, which affected disease diagnosis. Compared with the ASiR-V reconstruction algorithm, the DLIR reconstruction algorithm improved the image quality and diagnostic reliability for CCTA in patients with different weights, especially in heavier patients.

Noncanonical amino acids as doubly bio-orthogonal handles for one-pot preparation of protein multiconjugates.

Genetic encoding of noncanonical amino acid (ncAA) for site-specific protein modification has been widely applied for many biological and therapeutic applications. To efficiently prepare homogeneous protein multiconjugates, we design two encodable noncanonical amino acids (ncAAs), 4-(6-(3-azidopropyl)-s-tetrazin-3-yl) phenylalanine (pTAF) and 3-(6-(3-azidopropyl)-s-tetrazin-3-yl) phenylalanine (mTAF), containing mutually orthogonal and bioorthogonal azide and tetrazine reaction handles. Recombinant proteins and antibody fragments containing the TAFs can easily be functionalized in one-pot reactions with combinations of commercially available fluorophores, radioisotopes, PEGs, and drugs in a plug-and-play manner to afford protein dual conjugates to assess combinations of tumor diagnosis, image-guided surgery, and targeted therapy in mouse models. Furthermore, we demonstrate that simultaneously incorporating mTAF and a ketone-containing ncAA into one protein via two non-sense codons allows preparation of a site-specific protein triconjugate. Our results demonstrate that TAFs are doubly bio-orthogonal handles for efficient and scalable preparation of homogeneous protein multiconjugates.

Identification of oxidosqualene cyclases associated with saponin biosynthesis from Astragalus membranaceus reveals a conserved motif important for catalytic function.

INTRODUCTION: Triterpenoids and saponins have a broad range of pharmacological activities. Unlike most legumes which contain mainly oleanane-type scaffold, Astragalus membranaceus contains not only oleanane-type but also cycloartane-type saponins, for which the biosynthetic pathways are unknown. OBJECTIVES: This work aims to study the function and catalytic mechanism of oxidosqualene cyclases (OSCs), one of the most important enzymes in triterpenoid biosynthesis, in A. membranaceus. METHODS: Two OSC genes, AmOSC2 and AmOSC3, were cloned from A. membranaceus. Their functions were studied by heterologous expression in tobacco and yeast, together with in vivo transient expression and virus-induced gene silencing. Site-directed mutagenesis and molecular docking were used to explain the catalytic mechanism for the conserved motif. RESULTS: AmOSC2 is a ß-amyrin synthase which showed higher expression levels in underground parts. It is associated with the production of ß-amyrin and soyasaponins (oleanane-type) in vivo. AmOSC3 is a cycloartenol synthase expressed in both aerial and underground parts. It is related to the synthesis of astragalosides (cycloartane-type) in the roots, and to the synthesis of cycloartenol as a plant sterol precursor. From AmOSC2/3, conserved triad motifs VFM/VFN were discovered for ß-amyrin/cycloartenol synthases, respectively. The motif is a critical determinant of yield as proved by 10 variants from different OSCs, where the variant containing the conserved motif increased the yield by up to 12.8-fold. Molecular docking and mutagenesis revealed that Val, Phe and Met residues acted together to stabilize the substrate, and the cation-π interactions from Phe played the major role. CONCLUSION: The study provides insights into the biogenic origin of oleanane-type and cycloartane-type triterpenoids in Astragalus membranaceus. The conserved motif offers new opportunities for OSC engineering.

Two novel variants in a Bardet-Biedl syndrome type 5 patient with severe renal phenotype.

Bardet-Biedl syndrome type 5 (BBS5) has never been reported in Chinese populations. The aim of this study is to report the first BBS5 case in China, explore the phenotype and genotype correlation. The case was male, Han nationality, born with polydactyly and gained weight after birth, accompanied by polydipsia, polyuria and nocturia. He was found to have low vision at the age of 7 years, and having insufficient renal function at the age of 20 years. After hospitalization, he was found to have suffered from atrophy of the whole layer of macular retina, and end stage of kidney disease, presenting with shrinking and cyst-like changes of bilateral kidneys. Whole-exome sequencing was performed among the proband and his parents (Trios), further validated using Sanger sequencing and quantitative polymerase chain reaction. Two novel compound heterozygous variants of BBS5 gene [a missense variant NC_000002.12, NM_152384.3:c.1A>G(p.Met1?) & a large deletion c.(?_-60)_(386 + 1_387-1)del] were detected. BBS is rare, whereas BBS5 is rarer. Herein, we reported a Chinese BBS5 patient with severe renal phenotype and identified two novel BBS5 variants.

Deoxyhypusine hydroxylase as a novel pharmacological target for ischemic stroke via inducing a unique post-translational hypusination modification.

Ischemic stroke remains one of the leading causes of death worldwide, thereby highlighting the urgent necessary to identify new therapeutic targets. Deoxyhypusine hydroxylase (DOHH) is a fundamental enzyme catalyzing a unique posttranslational hypusination modification of eukaryotic translation initiation factor 5A (eIF5A) and is highly involved in the progression of several human diseases, including HIV-1 infection, cancer, malaria, and diabetes. However, the potential therapeutic role of pharmacological regulation of DOHH in ischemic stroke is still poorly understood. Our study first discovered a natural small-molecule brazilin (BZ) with an obvious neuroprotective effect against oxygen-glucose deprivation/reperfusion insult. Then, DOHH was identified as a crucial cellular target of BZ using HuProt™ human proteome microarray. By selectively binding to the Cys232 residue, BZ induced a previously undisclosed allosteric effect to significantly increase DOHH catalytic activity. Furthermore, BZ-mediated DOHH activation amplified mitophagy for mitochondrial function and morphology maintenance via DOHH/eIF5A hypusination signaling pathway, thereby protecting against ischemic neuronal injury in vitro and in vivo. Collectively, our study first identified DOHH as a previously unreported therapeutic target for ischemic stroke, and provided a future drug design direction for DOHH allosteric activators using BZ as a novel molecular template.

Nitazoxanide and related thiazolides induce cell death in cancer cells by targeting the 20S proteasome with novel binding modes.

Nitazoxanide and related thiazolides are a novel class of anti-infectious agents against protozoan parasites, bacteria and viruses. In recent years, it is demonstrated that thiazolides can also induce cell cycle arrest and apoptotic cell death in cancer cells. Due to their fast proliferating nature, cancer cells highly depend on the proteasome system to remove aberrant proteins. Many of these aberrant proteins are regulators of cell cycle progression and apoptosis, such as the cyclins, BCL2 family members and nuclear factor of κB (NF-κB). Here, we demonstrate at both molecular and cellular levels that the 20S proteasome is a direct target of NTZ and related thiazolides. By concurrently inhibiting the multiple catalytic subunits of 20S proteasome, NTZ promotes cell cycle arrest and triggers cell death in colon cancer cells, either directly or as a sensitizer to other anti-tumor agents, especially doxorubicin. We further show that the binding mode of NTZ in the ß5 subunit of the 20S proteasome is different from that of bortezomib and other existing proteasome inhibitors. These findings provide new insights in the design of novel small molecular proteasome inhibitors as anti-tumor agents suitable for solid tumor treatment in an oral dosing form.

Deep Learning Reconstruction Shows Better Lung Nodule Detection for Ultra-Low-Dose Chest CT.

Background Ultra-low-dose (ULD) CT could facilitate the clinical implementation of large-scale lung cancer screening while minimizing the radiation dose. However, traditional image reconstruction methods are associated with image noise in low-dose acquisitions. Purpose To compare the image quality and lung nodule detectability of deep learning image reconstruction (DLIR) and adaptive statistical iterative reconstruction-V (ASIR-V) in ULD CT. Materials and Methods Patients who underwent noncontrast ULD CT (performed at 0.07 or 0.14 mSv, similar to a single chest radiograph) and contrast-enhanced chest CT (CECT) from April to June 2020 were included in this prospective study. ULD CT images were reconstructed with filtered back projection (FBP), ASIR-V, and DLIR. Three-dimensional segmentation of lung tissue was performed to evaluate image noise. Radiologists detected and measured nodules with use of a deep learning-based nodule assessment system and recognized malignancy-related imaging features. Bland-Altman analysis and repeated-measures analysis of variance were used to evaluate the differences between ULD CT images and CECT images. Results A total of 203 participants (mean age ± standard deviation, 61 years ± 12; 129 men) with 1066 nodules were included, with 100 scans at 0.07 mSv and 103 scans at 0.14 mSv. The mean lung tissue noise ± standard deviation was 46 HU ± 4 for CECT and 59 HU ± 4, 56 HU ± 4, 53 HU ± 4, 54 HU ± 4, and 51 HU ± 4 in FBP, ASIR-V level 40%, ASIR-V level 80% (ASIR-V-80%), medium-strength DLIR, and high-strength DLIR (DLIR-H), respectively, of ULD CT scans (P < .001). The nodule detection rates of FBP reconstruction, ASIR-V-80%, and DLIR-H were 62.5% (666 of 1066 nodules), 73.3% (781 of 1066 nodules), and 75.8% (808 of 1066 nodules), respectively (P < .001). Bland-Altman analysis showed the percentage difference in long diameter from that of CECT was 9.3% (95% CI of the mean: 8.0, 10.6), 9.2% (95% CI of the mean: 8.0, 10.4), and 6.2% (95% CI of the mean: 5.0, 7.4) in FBP reconstruction, ASIR-V-80%, and DLIR-H, respectively (P < .001). Conclusion Compared with adaptive statistical iterative reconstruction-V, deep learning image reconstruction reduced image noise, increased nodule detection rate, and improved measurement accuracy on ultra-low-dose chest CT images. © RSNA, 2022 Online supplemental material is available for this article. See also the editorial by Lee in this issue.

O-Glycosylation Induces Amyloid-ß To Form New Fibril Polymorphs Vulnerable for Degradation.

Brain accumulation of amyloid-ß (Aß) peptides (resulting from a disrupted balance between biosynthesis and clearance) occurs during the progression of Alzheimer's disease (AD). Aß peptides have diverse posttranslational modifications (PTMs) that variously modulate Aß aggregation into fibrils, but understanding the mechanistic roles of PTMs in these processes remains a challenge. Here, we chemically synthesized three homogeneously modified isoforms of Aß (1-42) peptides bearing Tyr10 O-glycosylation, an unusual PTM initially identified from the cerebrospinal fluid samples of AD patients. We discovered that O-glycans significantly affect both the aggregation and degradation of Aß42. By combining cryo-EM and various biochemical assays, we demonstrate that a Galß1-3GalNAc modification redirects Aß42 to form a new fibril polymorphic structure that is less stable and more vulnerable to Aß-degrading enzymes (e.g., insulin-degrading enzyme). Thus, beyond showing how particular O-glycosylation modifications affect Aß42 aggregation at the molecular level, our study provides powerful experimental tools to support further investigations about how PTMs affect Aß42 fibril aggregation and AD-related neurotoxicity.

APEX2-based Proximity Labeling of Atox1 Identifies CRIP2 as a Nuclear Copper-binding Protein that Regulates Autophagy Activation.

Mammalian cell nuclei contain copper, and cancer cells are known to accumulate aberrantly high copper levels, yet the mechanisms underlying nuclear accumulation and copper's broader functional significance remain poorly understood. Here, by combining APEX2-based proximity labeling focused on the copper chaperone Atox1 with mass spectrometry we identified a previously unrecognized nuclear copper binding protein, Cysteine-rich protein 2 (CRIP2), that interacts with Atox1 in the nucleus. We show that Atox1 transfers copper to CRIP2, which induces a change in CRIP2's secondary structure that ultimately promotes its ubiquitin-mediated proteasomal degradation. Finally, we demonstrate that depletion of CRIP2-as well as copper-induced CRIP2 degradation-elevates ROS levels and activates autophagy in H1299 cells. Thus, our study establishes that CRIP2 as an autophagic suppressor protein and implicates CRIP2-mediated copper metabolism in the activation of autophagy in cancer cells.

Application of double layered end-to-end anastomosis with continuous manual suture for completing digestive tract reconstruction in totally laparoscopic distal gastrectomy.

BACKGROUND: We retrospectively reviewed and consecutively collected the clinical data of distal gastric cancer patients who received surgical treatment, and we discuss the safety and feasibility of double layered end-to-end anastomosis with continuous manual suture to complete digestive tract reconstruction in totally laparoscopic distal gastrectomy. METHODS: We reviewed the clinical data of 41 patients with distal gastric cancer from the gastroenterology department of the Second Affiliated Hospital of Dalian Medical University, from September 2018 to August 2019, who underwent totally laparoscopic distal gastrectomy. During the operation, the method of double layered end-to-end anastomosis with continuous manual suture was used for Billroth type I anastomosis to complete digestive tract reconstruction. All patients have been given a follow-up visit and gastroscopy three months after the operation. The peri-operative clinical information and postoperative follow-up information were collected for analysis, and the clinical application value was evaluated. RESULTS: General information: male(n = 27), female(n = 14), age = 65.02(SD 9.94) years, and BMI = 23.52(SD 2.56) kg/m2, Tumor location: antrum(32,78.0%), angle (6,14.6%), and body (3,7.3%). Clinical stage: I (27, 65.9%), II (7, 17.1%), and III (7, 17.1%). Operative information: operation time = 154.51(SD 33.37) min, anastomosis time = 26.88(SD 5.11) min; intraoperative bleeding = 66.34(SD 48.81) ml; first postoperative ambulation Median = 1(IQR 0) d, first postoperative flatus Median = 3(IQR 2) d, first postoperative diet Median = 3(IQR 1) d, postoperative hospital stay Median = 7(IQR 2) d, and total hospitalization cost = 10,935.00(SD 2205.72)USD. Differentiation degree: high and high-moderate (3,7.32%), moderate and poor-moderate (24, 58.54%), poor differentiation (14, 34.15%), dissected lymph nodes Median = 31(IQR 17), and positive lymph nodes Median = 0(IQR 1). Pathological stage: IA (20, 48.78%), IB (3, 7.32%), IIA (4, 9.76%), IIB (5, 12.20%), IIIA (1, 2.44%), IIIB (3, 7.32%), and IIIC (5, 12.20%). Complications (n = 4): lung infection (1, 2.44%), anastomotic leakage (1, 2.44%), and gastroparesis (2, 4.88%). CONCLUSION: It is safe and feasible in clinical treatment to apply the method of double layered end-to-end anastomosis with continuous manual suture to complete digestive tract reconstruction in totally laparoscopic distal gastrectomy.

Sudden onset of nephrotic syndrome in an asymptomatic Fabry patient: a case report.

BACKGROUND: Fabry disease (FD) is an X-linked lysosomal storage disorder caused by the mutation of the GLA gene, encoding the α-galactosidase, which is responsible for the catabolism of neutral glycosphingolipids. Microalbuminuria or low-grade proteinuria, and continuously progressive renal failure are common manifestations in FD males. However, sudden onset of nephrotic syndrome in FD, is rarely reported. CASE REPORT: A 32-year-old Chinese man was admitted to our hospital because of sudden onset of generalized edema due to nephrotic syndrome. He denied hypohidrosis, nocturia, and any history of episodic hand or foot pain. A few scattered angiokeratoma can be found on the low back skin on examination. Except for the similar locating pattern of angiokeratoma, no evident abnormality was found in the laboratory work up and physical examination of his younger brother. The patient was diagnosed with FD companying with minimal change disease by renal biopsy. Genetic analysis on our patient and his sibling revealed a nonsense GLA gene variant (c.707G > A, p.Trp236*), which has been previously reported in FD. Immunotherapy alone (steroids and tacrolimus), but without enzyme replacement therapy, much improved the massive proteinuria. Follow up to date, his 24-h urine protein is stable at about 0.5 g, and renal function keeps normal. CONCLUSION: Sudden onset of nephrotic syndrome, although rare, may occur in FD, even as the primary renal manifestation, but this usually suggests additional renal disease. Immunosuppressive treatment should be considered in such FD patient companying with nephrotic syndrome.