Deleterious ZNRF3 germline variants cause neurodevelopmental disorders with mirror brain phenotypes via domain-specific effects on Wnt/ß-catenin signaling.

Zinc and RING finger 3 (ZNRF3) is a negative-feedback regulator of Wnt/ß-catenin signaling, which plays an important role in human brain development. Although somatically frequently mutated in cancer, germline variants in ZNRF3 have not been established as causative for neurodevelopmental disorders (NDDs). We identified 12 individuals with ZNRF3 variants and various phenotypes via GeneMatcher/Decipher and evaluated genotype-phenotype correlation. We performed structural modeling and representative deleterious and control variants were assessed using in vitro transcriptional reporter assays with and without Wnt-ligand Wnt3a and/or Wnt-potentiator R-spondin (RSPO). Eight individuals harbored de novo missense variants and presented with NDD. We found missense variants associated with macrocephalic NDD to cluster in the RING ligase domain. Structural modeling predicted disruption of the ubiquitin ligase function likely compromising Wnt receptor turnover. Accordingly, the functional assays showed enhanced Wnt/ß-catenin signaling for these variants in a dominant negative manner. Contrarily, an individual with microcephalic NDD harbored a missense variant in the RSPO-binding domain predicted to disrupt binding affinity to RSPO and showed attenuated Wnt/ß-catenin signaling in the same assays. Additionally, four individuals harbored de novo truncating or de novo or inherited large in-frame deletion variants with non-NDD phenotypes, including heart, adrenal, or nephrotic problems. In contrast to NDD-associated missense variants, the effects on Wnt/ß-catenin signaling were comparable between the truncating variant and the empty vector and between benign variants and the wild type. In summary, we provide evidence for mirror brain size phenotypes caused by distinct pathomechanisms in Wnt/ß-catenin signaling through protein domain-specific deleterious ZNRF3 germline missense variants.

Development of an Integrated Disposable Device for SARS-CoV-2 Nucleic Acid Extraction and Detection.

Objective: To develop a highly sensitive and rapid nucleic acid detection method for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Methods: We designed, developed, and manufactured an integrated disposable device for SARS-CoV-2 nucleic acid extraction and detection. The precision of the liquid transfer and temperature control was tested. A comparison between our device and a commercial kit for SARS-Cov-2 nucleic acid extraction was performed using real-time fluorescence reverse transcription polymerase chain reaction (RT-PCR). The entire process, from SARS-CoV-2 nucleic acid extraction to amplification, was evaluated. Results: The precision of the syringe transfer volume was 19.2 ± 1.9 µL (set value was 20), 32.2 ± 1.6 (set value was 30), and 57.2 ± 3.5 (set value was 60). Temperature control in the amplification tube was measured at 60.0 ± 0.0 °C (set value was 60) and 95.1 ± 0.2 °C (set value was 95) respectively. SARS-Cov-2 nucleic acid extraction yield through the device was 7.10 × 10 6 copies/mL, while a commercial kit yielded 2.98 × 10 6 copies/mL. The mean time to complete the entire assay, from SARS-CoV-2 nucleic acid extraction to amplification detection, was 36 min and 45 s. The detection limit for SARS-CoV-2 nucleic acid was 250 copies/mL. Conclusion: The integrated disposable devices may be used for SARS-CoV-2 Point-of-Care test (POCT).

Nursing Management of Adult Ostomy Patients During the COVID-19 Pandemic.

The continuation of the COVID-19 epidemic poses novel challenges for adult ostomy patients care. It is essential to explore nursing management for ostomy patients to ensure patients receive standardized care while minimizing exposure to COVID-19. This article reviews the perioperative nursing of Chinese adult ostomy patients in the post-epidemic era, as well as outpatient review after discharge and home care, to provide reference and basis for medical staff and patients in the post-pandemic era.

Case Report: Clinical characteristics and genetic analysis of two patients with hereditary hemorrhagic telangiectasia.

Objective: To analyze the clinical features and genetic characteristics of two patients with hereditary hemorrhagic telangiectasia (HHT) and to review the relevant literature. Methods: The clinical data of two HHT patients admitted to the author's hospital between April 2019 and February 2022 were retrospectively analyzed. Meanwhile, the genetic analysis was performed with their consent. Results: The first patient was a 62-year-old woman who had been complaining of shortness of breath and fever for 20 days. Her previous medical history included brain abscess drainage and video-assisted thoracoscopic surgery for a pulmonary hemangioma. A right heart catheterization revealed no pulmonary arterial hypertension, and an abdominal enhanced magnetic resonance imaging revealed multiple arteriovenous malformations in the liver. Her ACVRL1 heterozygous variants were discovered through whole-exon gene testing. The second case involved a 47-year-old woman who had been experiencing chest tightness for the past 2 years. Several years ago, she underwent brain abscess drainage and embolization of a pulmonary arteriovenous fistula. Ultrasound revealed generalized hepatic vascular dilation, and enhanced computed tomography revealed numerous pulmonary venous fistulas scattered in both lungs as well as multiple arteriovenous malformations in the liver. Her whole-exon gene testing revealed that she, like her son, had heterozygous ENG variants. Conclusion: HHT patients may experience infection, bleeding, dyspnea, and other symptoms. Imaging is important in disease diagnosis and management because early detection and treatment can prevent major complications and disability or even death.

Morphology of reproductive accessory glands in female Sepia pharaonis (Cephalopoda: Sepiidae) sheds light on egg encapsulation.

The pharaoh cuttlefish, Sepia pharaonis, is an important cephalopod fishery species in southeastern Asia, with understudied reproductive physiology. The present study aimed to investigate the cellular characteristics of epithelial cells found in the nidamental glands (NGs) and accessory NGs (ANGs), as well as the structural connections between these two glands in mature female S. pharaonis. A histological analysis revealed two types of epithelial cells in NGs: Alcian blue-positive, PAS-negative mucosubstance-secreting cells and eosinophilic, PAS-positive granule-secreting cells. Using transmission electron microscopy, three types of epithelial cells were identified: cells with electron-dense granules, cells with electron-lucent granules, and cells with both cilia and microvilli in the apex. Mature ANGs contain an abundance of tubular units composed of epithelial cells resting on a thin layer of basal lamina. Innervated muscle cells are tightly adhered to the basal lamina. In addition, we observed epithelial canalization of ANG tubules penetrating through the connective tissue linking NGs and the walls of the tubules in ANGs, which allows the contents of the ANG tubules to be transported to the NGs. Our results suggest that ANGs participate in the encapsulation of the ova via the same pathway as NGs, which provides an important basis for future studies on the mechanism of protection provided by NGs and ANGs during embryonic development in S. pharaonis.

[Effects of butylphthalide on bronchial asthma in guinea pigs and involvement of endothelin].

OBJECTIVE: To study the effects of butylphthalide on bronchial asthma in guinea pigs, and investigate the involvement of endothelin. METHODS: In guinea pigs, bronchial asthma was induced by injection of ovalbumin (OVA) and provoked by inhalation of OVA, and the effects of butylphthalide on asthma were evaluated through the changes it induced by OVA, pulmonary function, endothelin-1 (ET-1) contents and activity of endothelin converting enzyme-1 (ECE-1) in bronchoalveolar lavage fluid (BALF), serum and lung tissue, and the gene expression of ET-1 in lung tissue. RESULTS: Butylphthalide significantly improved pulmonary function, lowered asthmatic behavior score, inhibited the activity of ECE-1, and reduced ET-1 gene expression level in lung tissue. CONCLUSIONS: Butylphthalide has an anti-asthma effect and the mechanisms involve inhibition of ECE-1 activity and lowering of ET-1geng expression.

Development of the genital duct system in the protandrous black porgy, Acanthopagrus schlegeli.

Protandrous black porgies, Acanthopagrus schlegeli, have a striking life cycle, which includes early sex differentiation, bisexual gonads, and a male-to-female sex change at three years of age. We found novel features of and insights into the development of the genital duct system in relation to the gonadal stage during early gonadal development and natural sex change. We found that the genital ducts developed at 16-20 weeks of age during sex differentiation. The gonad developed "ovarian cavity cracks" and became "four-stranded" during the first prespawning period and then proceeded to the development of genital ducts before 1 year of age. Two ovarian cavities of the paired gonads combined, developed, and extended caudally to form the oviduct, making up the inner duct of the genital ducts. The testicular main cavities also extended and fused together to form the outer duct of the genital duct system, that is, the sperm duct. The coexistence of an outer sperm duct and an inner oviduct constituted a unique "double cannula genital duct" structure. Gradually the inferior walls of the oviduct intermingled with those of the sperm duct, and the circular lumen of the sperm duct changed into an "M-shaped canal." Finally, the sperm duct and oviduct separated completely at the distal part of genital duct system. During natural sex change, the male reproductive passage regressed and degenerated and was replaced by connective tissue. The oviduct arrested as a blunt end during male phase and, finally, extended and connected to the genital pore during the female phase.

In situ real-time investigation of cancer cell photothermolysis mediated by excited gold nanorod surface plasmons.

The photothermolysis of living EMT-6 breast tumor cells triggered by gold nanorods (AuNRs) with two-photon irradiation was conducted in situ and under real-time observation. The morphology and plasma membrane permeability of the cells were key indicators to the phenomena. AuNRs with an aspect ratio of 3.92, and a longitudinal absorption peak at 800 nm were synthesized with a seed-mediated method. The nanorods surfaces were further modified with polystyrenesulfonate (PSS) for biocompatibility. The prepared nanorods displayed excellent two-photon photoluminescence imaging. In situ real-time results revealed cavities internal to the cells were created from thermal explosions triggered by AuNRs localized photothermal effect. The cavitation dynamic is energy dependent and responsible for the perforation or sudden rupture of the plasma membrane. The energy threshold for cell therapy depended significantly on the number of nanorods taken up per cell. For an ingested AuNR cluster quantity N approximately 10-30 per cell, it is found that energy fluences E larger-than 93 mJ/cm(2) led to effective cell destruction in the crumbled form within a very short period. As for a lower energy level E = 18 mJ/cm(2) with N approximately 60-100, a non-instant, but progressive cell deterioration, is observed.

Effect of estrogen on the activity and growth of human osteoclasts in vitro.

OBJECTIVE: Estrogen deficiency results in postmenopausal osteoporosis by increasing the rate of bone loss. The mechanism responsible for the effects of estrogen on osteoclasts is still unclear. MATERIALS AND METHODS: The potential of mononuclear cells from cord blood or bone marrow to differentiate into mature osteoclasts when co-cultured with human osteoblast cells was investigated. The effects of estrogen on osteoclastogenesis and osteoclast activity were also examined. RESULTS: Macrophage markers CD11b and CD14 were downregulated and vitronectin receptor was upregulated during 28 days' co-culture of mononuclear cells and human osteoblasts. Long-term co-culture resulted in the formation of numerous large tartrate-resistant acid phosphatase-positive multinucleated cells capable of resorption of bone slices. After incubation for 28 days, the addition of 17beta-estradiol caused a significant decrease in the expression of vitronectin receptor and tartrate-resistant acid phosphatase-positive multinucleated cells in cultures derived from both bone marrow and cord blood. A significant decrease in bone resorption was also noted in the presence of estrogen. CONCLUSION: Estrogen not only suppresses osteoclastogenesis but also inhibits the activity of osteoclasts.

Estradiol-17beta triggers luteinizing hormone release in the protandrous black porgy (Acanthopagrus schlegeli Bleeker) through multiple interactions with gonadotropin-releasing hormone control.

We investigated the mechanism of estradiol-17beta (E2) action on stimulation of LH (=gonadotropin II) release in the black porgy fish (Acanthopagrus schlegeli Bleeker) using an in vivo approach and primary cultures of dispersed pituitary cells in vitro. In vivo, E2 but not androgens (testosterone [T] and 11-ketotestosterone [11-KT]) significantly stimulated plasma LH in a dose-dependent manner. Estradiol-17beta also increased brain content of seabream GnRH. GnRH antagonist prevented E2 stimulation of LH release in vivo, indicating that the effect of E2 on LH was mediated by GnRH. In vitro, sex steroids (E2, T, 11-KT) alone had no effect on basal LH release in the cultured pituitary cells, but GnRH significantly stimulated LH release. Estradiol-17beta potentiated GnRH stimulation of LH release, an effect that was inhibited by GnRH antagonist, and 11-KT, but not T, also potentiated GnRH stimulation of LH release. The potentiating effect of 11-KT on GnRH-induced LH release in vitro was stronger than that of E2. These data suggest that E2 triggers LH release in vivo by acting both on GnRH production at the hypothalamus and on GnRH action at the pituitary. In contrast, 11-KT may only stimulate GnRH action at the pituitary. The E2) induction of LH release, through multiple interactions with GnRH control, supports a possible central role of E2in the sex change observed in the protandrous black porgy.