The elevated polyamines, amine-rich molecules with diverse functions in pathophysiology processes, are implicated in contributing to tumorigenesis and progression. Whether and how they affect the efficacy of chemotherapy is incompletely understood. Our screening assays reveal that the supplement with a low dose of spermidine (Spd), one of the polyamines, enhances ferroptosis in prostate cancer cells as evidenced by increased lipid peroxidation and intracellular Fe²⁺ levels in vitro. Combination treatment with Spd and a low dose of ferroptosis inducer erastin synergistically augments anti-tumor efficacy with undetectable toxicity in mice. Analysis of RNA-seq data indicates that heme oxygenase 1 (HMOX1), an enzyme that catalyzes the cleavage of heme to release Fe²⁺, is significantly upregulated in response to Spd and erastin cotreatment. Spd mediated the hypusine modification of the eukaryotic initiation factor 5A (EIF5A) promotes the translation of the nuclear factor erythroid 2-related factor 2 (NRF2), subsequently leading to elevation of HMOX1. Moreover, Spd and erastin significantly inhibit proteasome activity which results in a decrease in proteasomal degradation of NRF2, although many proteasome-related genes are induced either by Spd or Spd plus erastin. Thus, in addition to its pro-oncogenic activity, the supplement of Spd improves antitumor activity in combination with ferroptosis inducers and offers an optional approach to cancer treatment.

Sexual dimorphism in the brain underlies behavioral differences between sexes. The bed nucleus of the stria terminalis (BNST) is a complex nucleus that differs between males and females, but the sexual dimorphism in cytoarchitecture and the connectome of its oval subdivision (ovBNST) remains largely unexplored. By combining snRNA-seq and transgenic labeling, we found a higher density of ovBNST proenkephalin (ovBNST^PENK) neurons in male than female mice. Anatomically, we virally mapped the efferents and afferents of ovBNST^PENK neurons, finding reciprocally dimorphic connections with the hypothalamus and striatum. Gene enrichment analysis suggests that ovBNST^PENK neurons are modulated by the upstream dopamine pathway. Functionally, by applying caspase-3-mediated depletion of ovBNST^PENK neurons, we found that loss of these neurons enhanced locomotor activity in male but not female mice, without altering the anxiety-like phenotypes in either sex. Our study may pave the way for a better understanding of the anatomical and functional profiles of ovBNST^PENK neurons from a sexually dimorphic perspective.
Animals ; Male ; Female ; Septal Nuclei/physiology* ; Sex Characteristics ; Neurons/physiology* ; Enkephalins/metabolism* ; Mice ; Mice, Transgenic ; Protein Precursors/metabolism* ; Mice, Inbred C57BL ; Neural Pathways/physiology*

Objective:To provide a basis for its diagnosis and treatment for clinicians by analyzing the clinical manifestations, imaging features and surgical efficacy of primary intraspinal paraganglioma.Methods:The clinical data of 6 patients with intraspinal paraganglioma from April 2014 to January 2021 in Xiangyang Central Hospital were retrospectively analyzed, and all patients were treated with microsurgery via a posterior median approach.Results:All 6 patients achieved total tumor resection, and the postoperative pathological diagnosis was paraganglioma. Among them, 1 patient′s tumor located inside and outside the cervical spinal canal without destruction of the vertebral body; 1 patient′s tumor located lumbosacral canal with destruction of the vertebral body; the others 4 patients′ tumor located within the lumbar spinal canal. The patients were followed up for 12 to 120 months after surgery, with a median follow-up time of 61.5 months. MRI examination was performed at the last follow-up, and no recurrence was observed. The patients underwent MRI examination at the last follow-up, and none of the patients recurred.Conclusions:The intraspinal paraganglioma is a rare tumor, and nonfunctional benign tumors are predominant. Its clinical and imaging manifestations lack specificity and are often difficult to diagnose before surgery. Surgical resection, especially complete resection, has a better prognosis, and the effectiveness of adjuvant therapy is uncertain.

Objective:To establish a digital droplet RT-PCR(dRT-PCR) method for Hepatitis A virus (HAV), and compare it with Real time RT-PCR(RT-qPCR) method, and select the best method for detecting hepatitis A virus in strawberry.Methods:Extract HAV vaccine RNA, optimize the reaction conditions of dRT-PCR and evaluate its specificity; Alkaline elution -PEG concentration method was used to extract nucleic acid from strawberry samples. At the same time, dRT-PCR and RT-qPCR method were used to detect the sensitivity and inhibition rate of HAV vaccine RNA in pure water and strawberry matrix, and the recovery rate of HAV in artificially contaminated strawberry was compared, which was applied to the detection of commercially available samples.Results:The optimal annealing temperature for dRT-PCR reaction was 60 ℃, and the optimal concentrations of primers and probes were 0.4 μmol/L、0.4 μmol/L and 0.2 μmol/L, with good specificity. There is no significant difference in sensitivity between the two method in detecting HAV vaccine RNA in pure water and strawberry matrix. The inhibition rate of dRT-PCR is low. The recovery rates of dRT-PCR and RT-qRCR in the detection of strawberry samples contaminated with HAV at higher concentrations were 12.90±0.006% and 30.12±0.02%, respectively. The recovery rates of lower concentrations of HAV contaminated strawberry samples were 18.27±0.07% and 10.85±0.03%, respectively, and the difference was statistically significant ( P<0.05). When strawberry samples on the market were tested, the result of both method were negative. Conclusions:The sensitivity of dRT-PCR method established in this study is not significantly different from that of RT-qPCR in detecting HAV RNA in different substrates, but dRT-PCR has good tolerance to PCR reaction inhibitors and high recovery rate when detecting low concentration HAV. Both detection method can be used for quantitative detection of hepatitis A virus in strawberry, and can be selected according to the actual situation.

Objective:To explore the marking method for magnetically controlled capsule gastroscopy (MCCG) pictures with artificial intelligence (AI), so as to improve the work efficiency of endoscopist and to reduce the blind area of AI image reading.Methods:According to the consensus of MCCG, 24 parts of stomach in 14 775 pictures of MCCG from 35 subjects in Shenzhen Zifu Medical Technology Co., Ltd received MCCG from March to August, 2020 were marked by ten gastroenterologists and one developer of MCCG with medical background, the marking shape included rectangles and polygons. Among the ten gastroenterologists, three were senior endoscopist (the total number of gastroenteroscopy operations over 80 000, chief physician or associate chief physician), four were medium seniority endoscopist (the total number of gastroenteroscopy operations between 10 000 and 80 000, associate chief physician), and three were junior endoscopist (the total number of gastroenteroscopy operations less than 10 000, attending physician). The pictures of the same subject were pre-marked by two selected senior endoscopists with blind method, and the standard of marking with most appropriate coincidence rate was determined. The qualified marked pictures were automatically learn with AI deep learning method, and the learning results were fed back. Chi square test was used for statistical analysis.Results:According to the pre-marked results, the standard of coincidence rate for rectangular marking area was set as 50.0% and that for polygon marking area was 70.0%. The first correction for qualified rate was 39.0% (5 762/14 775). A total of 9 013 pictures were corrected. After repeated training and correction for one to five times, all pictures were qualified marked. The marking qualified rate of senior endoscopist partners was higher than that of partners of different qualifications (48.7%, 1 200/2 466 vs. 19.0%, 825/4 337), and the difference was statistically significant ( χ2=659.20, P<0.001). There was no statistically significant difference in the marking qualified rate between the senior endoscopist partners and partners of senior endoscopist and capsule developer (48.7%, 1 200/2 466 vs. 49.6%, 1 496/3 019; P>0.05). Conclusions:Establishment of AI marking method for MCCG can provide technical support for AI non-blind area reading, and AI non-blind area monitoring during the operation of MCCG.

Objective: To detect WNT10A gene mutations in patients with oligodontia or anodontia (≥6 teeth missing) and analyze their dental phenotype. Methods: Patients with oligodontia or anodontia were enrolled from the clinic for oral examination, genetic history collection and whole exon sequencing, and patients with WNT10A gene mutations were included. Sanger sequencing was utilized to validate the WNT10A gene variations in probands and family members compared with the normal sequence. The pathogenicity of WNT10A mutations was evaluated by functional prediction, conservation analysis and structure prediction of protein mutants. Implant rehabilitation was applied to restore the patients' oral function. Results: Five WNT10A gene mutations were detected in six unrelated patients, and c.26G>A (p. Trp9X) and c.1036delT (p. Cys346fs) were novel mutations with pathogenic potential. The mean number of missing teeth was (15.33±8.64) per case. The most frequently missing permanent teeth were maxillary canines (100%), and the least frequently missing teeth were mandibular first molars (25%). Implant rehabilitation was applied in five patients, and patients were found to have ideal implant osseointegration and functional restoration. Conclusion This study identified novel WNT10A gene pathogenic variants, enriching the WNT10A gene spectrum and providing new evidence for genetic diagnosis and prenatal consultation. Implant rehabilitation was also proven to be a treatment option for these patients.

Objective:To observe the ratio of helper T cells 17 (Th17)/regulatory T cells (Treg) in peripheral blood of patients with Hashimoto's thyroiditis (HT) and the expression changes of related cytokines in serum, and to explore their role in the occurrence and development of HT.Methods:Using the case-control study method, 35 HT patients examined in the General Hospital of Heilongjiang Beidahuang Group from February to November 2019 were selected as HT group, and 39 healthy people in the same period were selected as control group. Early morning fasting venous blood samples of the two groups were collected to test the levels of thyroid stimulating hormone (TSH), free thyroxine (FT 4), thyroid peroxidase antibody (TPOAb) and thyroglobulin antibody (TgAb). The expressions of serum interleukin (IL)-6, IL-17 and transforming growth factor β (TGF-β) were tested by enzyme-linked immunosorbent assay (ELISA); the number of Th17, Treg in peripheral blood were determined by flow cytometry. Results:The levels of TPOAb, TgAb and TSH in HT group [130.60 (43.37, 714.40), 368.10 (136.90, 1 103.00) U/ml, 9.05 (6.62, 15.23) μU/ml] were significantly higher than those in control group [2.66 (1.52, 4.69), 12.63 (11.43, 14.60) U/ml, 1.87 (1.36, 2.23) μU/ml, U = 6.87, 6.62, 4.85, P < 0.001], and the FT 4 level [0.76 (0.63, 1.04) ng/dl] was lower than that in control group [1.14 (1.02, 1.26) ng/dl, U = 7.39, P < 0.001]. The expressions of IL-6, IL-17 and TGF-β in HT group were higher than those in control group ( t = 2.41, 9.04, 2.44, P < 0.05). The number of Th17 and the ratio of Th17/Treg in HT group were higher than those in control group ( t = 4.20, 3.50, P < 0.05), and the number of Treg was lower than that in control group ( t = 4.45, P = 0.001). Conclusion:In HT patients, Th17 are increased, Treg decreased, Th17/Treg ratio increased, and the expressions of IL-6, IL-17 and TGF-β are increased.

Oligodontia is the congenital absence of six or more teeth and comprises the more severe forms of tooth agenesis. Many genes have been implicated in the etiology of tooth agenesis, which is highly variable in its clinical presentation. The purpose of this study was to identify associations between genetic mutations and clinical features of oligodontia patients. An online systematic search of papers published from January 1992 to June 2021 identified 381 oligodontia cases meeting the eligibility criteria of causative gene mutation, phenotype description, and radiographic records. Additionally, ten families with oligodontia were recruited and their genetic etiologies were determined by whole-exome sequence analyses. We identified a novel mutation in WNT10A (c.99_105dup) and eight previously reported mutations in WNT10A (c.433 G > A; c.682 T > A; c.318 C > G; c.511.C > T; c.321 C > A), EDAR (c.581 C > T), and LRP6 (c.1003 C > T, c.2747 G > T). Collectively, 20 different causative genes were implicated among those 393 cases with oligodontia. For each causative gene, the mean number of missing teeth per case and the frequency of teeth missing at each position were calculated. Genotype-phenotype correlation analysis indicated that molars agenesis is more likely linked to PAX9 mutations, mandibular first premolar agenesis is least associated with PAX9 mutations. Mandibular incisors and maxillary lateral incisor agenesis are most closely linked to EDA mutations.
Humans ; Phenotype ; Wnt Proteins

Although DNA 5-hydroxymethylcytosine (5hmC) is recognized as an important epige-netic mark in cancer, its precise role in lymph node metastasis remains elusive. In this study, we investigated how 5hmC associates with lymph node metastasis in breast cancer. Accompanying with high expression of TET1 and TET2 proteins, large numbers of genes in the metastasis-positive pri-mary tumors exhibit higher 5hmC levels than those in the metastasis-negative primary tumors. In contrast, the TET protein expression and DNA 5hmC decrease significantly within the metastatic lesions in the lymph nodes compared to those in their matched primary tumors. Through genome-wide analysis of 8 sets of primary tumors, we identified 100 high-confidence metastasis-associated 5hmC signatures, and it is found that increased levels of DNA 5hmC and gene expression of MAP7D1 associate with high risk of lymph node metastasis. Furthermore, we demonstrate that MAP7D1, regulated by TET1, promotes tumor growth and metastasis. In conclusion, the dynamic 5hmC profiles during lymph node metastasis suggest a link between DNA 5hmC and lymph node metastasis. Meanwhile, the role of MAP7D1 in breast cancer progression suggests that the metastasis-associated 5hmC signatures are potential biomarkers to predict the risk for lymph node metastasis, which may serve as diagnostic and therapeutic targets for metastatic breast cancer.