Application and insights of targeted next-generation sequencing in a large cohort of 46,XY disorders of sex development in Chinese.

PURPOSE: 46,XY disorders of sex development (46,XY DSD) are characterized by incomplete masculinization of genitalia with reduced androgenization. Accurate clinical management remains challenging, especially based solely on physical examination. Targeted next-generation sequencing (NGS) with known pathogenic genes provides a powerful tool for diagnosis efficiency. This study aims to identify the prevalent genetic variants by targeted NGS technology and investigate the diagnostic rate in a large cohort of 46,XY DSD patients, with most of them presenting atypical phenotypes. METHODS: Two different DSD panels were developed for sequencing purposes, targeting a cohort of 402 patients diagnosed with 46,XY DSD, who were recruited from the Department of Urology at Children's Hospital, Zhejiang University School of Medicine (Hangzhou, China). The detailed clinical characteristics were evaluated, and peripheral blood was collected for targeted panels to find the patients' variants. The clinical significance of these variants was annotated according to American College of Medical Genetics and Genomics (ACMG) guidelines. RESULTS: A total of 108 variants across 42 genes were found in 107 patients, including 46 pathogenic or likely pathogenic variants, with 45.7%(21/46) being novel. Among these genes, SRD5A2, AR, FGFR1, LHCGR, NR5A1, CHD7 were the most frequently observed. Besides, we also detected some uncommon causative genes like SOS1, and GNAS. Oligogenic variants were also identified in 9 patients, including several combinations PROKR2/FGFR1/CYP11B1, PROKR2/ATRX, PROKR2/AR, FGFR1/LHCGR/POR, FGFR1/NR5A1, GATA4/NR5A1, WNT4/AR, MAP3K1/FOXL2, WNT4/AR, and SOS1/FOXL2. CONCLUSION: The overall genetic diagnostic rate was 11.2%(45/402), with an additional 15.4% (62/402) having variants of uncertain significance. Additionally, trio/duo patients had a higher genetic diagnostic rate (13.4%) compared to singletons (8.6%), with a higher proportion of singletons (15.1%) presenting variants of uncertain significance. In conclusion, targeted gene panels identified pathogenic variants in a Chinese 46,XY DSD cohort, expanding the genetic understanding and providing evidence for known pathogenic genes' involvement.

46,XY disorders of sex development (46,XY DSD) are conditions where individuals don't fully develop male genitalia due to reduced androgen hormones. Diagnosing these conditions based only on physical exams is difficult. This study used advanced genetic testing called targeted next-generation sequencing (NGS) to identify common genetic variations in a large group of 46,XY DSD patients, many of whom had unusual symptoms. We examined 402 patients with DSD and a 46,XY karyotype, focusing on 142 candidate genes related to sex development. We found genetic variations in 107 patients, including 45 that were likely responsible for their condition. Some of these variations were new discoveries. The most commonly affected genes were SRD5A2, AR, FGFR1, LHCGR, NR5A1, CHD7. We also found that some patients had variations in multiple genes, suggesting complex genetic causes. Overall, we were able to diagnose 11.2% of patients based on our genetic testing, with another15.4% having uncertain results. Patients tested as a trio or duo (with their parents) had a higher diagnosis rate than those tested alone. This study helps expand our understanding of the genetic factors behind 46,XY DSD in the Chinese population.

Fabrication of a Janus Copper Mesh by SiO2 Spraying for Unidirectional Water Transportation and Oil/Water Separation.

Massive discharge of oily wastewater and frequent occurrence of offshore oil spills have posed an enormous threat to the socioeconomic and ecological environments. Janus membranes with asymmetric wettability properties have great potential for oil/water separation applications and have attracted widespread attention. However, existing Janus membranes still suffer from complex and costly manufacturing processes, low permeability, and poor recyclability. Herein, a novel and facile strategy was proposed to fabricate a Janus copper mesh with opposite wettability for unidirectional water transport and efficient oil/water separation. The hydrophilic side of the Janus copper mesh was prepared by coating it with Cu(OH)2 nanoneedles via a chemical oxidation method. The hydrophobic side was fabricated by coating it with hydrophobic SiO2 nanoparticles via a facile spraying method. The as-prepared Janus copper mesh showed asymmetric surface wettability, which can achieve unidirectional water transport and efficient oil/water separation with excellent recyclability, exhibiting great application potential for droplet manipulation and wastewater purification.

Liquid-Assisted Bionic Conical Needle for In-Air and In-Oil-Water Droplet Ultrafast Unidirectional Transportation and Efficient Fog Harvesting.

Learning from nature, many bionic materials and surfaces have been developed for the directional transportation of water and fog collection. However, current research mainly focuses on the self-transportation behavior of droplets in air-phase environments, rarely concerning underoil environments. Herein, in this work, a liquid-assisted bionic copper needle was fabricated for the rapid self-transportation of water droplets in air and oil environments. The water droplet can be spontaneously transported on the as-prepared bionic copper needle from the tip to the base. More importantly, the water-prewetted bionic copper needle can achieve the ultrafast unidirectional transportation of a water droplet in an oil environment, showing a maximum transport velocity of 76.2 mm/s and a transport distance over 33 mm, which were ten times higher than those reported in the previous research. Additionally, the droplet transport mechanism was revealed. The effects of the apex angle and tilt angle of the as-prepared bionic needle and droplet volume on the self-transportation behavior of water droplets were systematically investigated. The results indicated that the transport velocity of the water droplet decreased with the increase of the apex angle of the conical needle, while it increased with the increase of the droplet volume and needle tilt angle. Furthermore, the as-prepared bionic copper needle exhibited excellent fog collection performance with a single copper needle fog collecting efficiency of up to 2220 mg/h, which was 9.7 times that of the original copper needle. In summary, this work provides a simple and novel method to fabricate bionic copper needles for the directional self-transportation of water droplets in air-phase and oil-phase environments as well as efficient fog collection. It shows great application potential in the fields of microfluidics, desalination, and freshwater collection.

Preparation of a Janus copper mesh via nanoparticle interface self-assembly for unidirectional water transportation.

A simple colloidal particle interface assembly method is presented to fabricate Janus meshes. The transport mechanism of water on the Janus mesh was fully revealed by experimental observation and numerical simulation. Furthermore, a liquid-assisted ultrafast transport of water droplets on the Janus mesh was presented (transport speed was increased by more than 20 times).

Heat Shock Transcription Factor GhHSFB2a Is Crucial for Cotton Resistance to Verticillium dahliae.

Heat shock transcription factors (HSFs) play a critical regulatory role in many plant disease resistance pathways. However, the molecular mechanisms of cotton HSFs involved in resistance to the soil-borne fungus Verticillium dahliae are limited. In our previous study, we identified numerous differentially expressed genes (DEGs) in the transcriptome and metabolome of V. dahliae-inoculated Arabidopsis thaliana. In this study, we identified and functionally characterized GhHSFB2a, which is a DEG belonging to HSFs and related to cotton immunity to V. dahliae. Subsequently, the phylogenetic tree of the type two of the HSFB subfamily in different species was divided into two subgroups: A. thaliana and strawberry, which have the closest evolutionary relationship to cotton. We performed promoter cis-element analysis and showed that the defense-reaction-associated cis-acting element-FC-rich motif may be involved in the plant response to V. dahliae in cotton. The expression pattern analysis of GhHSFB2a displayed that it is transcriptional in roots, stems, and leaves and significantly higher at 12 h post-inoculation (hpi). Subcellular localization of GhHSFB2a was observed, and the results showed localization to the nucleus. Virus-induced gene silencing (VIGS) analysis exhibited that GhHSFB2a silencing increased the disease index and fungal biomass and attenuated resistance against V. dahliae. Transcriptome sequencing of wild-type and GhHSFB2a-silenced plants, followed by Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, protein-protein interaction, and validation of marker genes revealed that ABA, ethylene, linoleic acid, and phenylpropanoid pathways are involved in GhHSFB2a-mediated plant disease resistance. Ectopic overexpression of the GhHSFB2a gene in Arabidopsis showed a significant increase in the disease resistance. Cumulatively, our results suggest that GhHSFB2a is required for the cotton immune response against V. dahliae-mediated ABA, ethylene, linoleic acid, and phenylpropanoid pathways, indicating its potential role in the molecular design breeding of plants.

Whether the Unilateral Transverse Process-pedicle Approach has Advantages over the Traditional Transpedicle Approach: A Systematic review and Meta-analysis.

PURPOSE: To summarize the literature and compare the advantages and disadvantages of the unilateral transverse process-pedicle approach (UTPA) and conventional transpedicular approach (CTPA) vertebral augmentation in the treatment of osteoporotic vertebral compression fractures (OVCF). METHODS: A single researcher performed a systematic literature review using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Online scientific databases were searched in September 2021 for English- and Chinese-language publications. A series of comparative studies were included, with UTPA as the main intervention and CTPA as the comparison indicator. A meta-analysis was performed for studies that reported clinical outcome indicators. The χ2 was used to study heterogeneity between trials, and the I2 statistic was calculated to estimate variation across studies. RESULTS: A total of eight studies were included for meta-analysis, all of which were observational studies with mixed bias risk. There were 613 subjects in the UTPA group and 488 subjects in the CTPA group. The results of the meta-analysis showed that there was no difference between the UTPA group and the CTPA group in terms of visual analogue scale scores (p = 0.31), Oswestry Disability Index scores (p = 0.50), correction of kyphosis angle (p = 0.65), and the amount of bone cement (p = 0.13), but the UTPA group had a shorter operative time (p < 0.001), bone cement leakage rates (p = 0.02), and fluoroscopy times than the CTPA group (p < 0.001). Partial analysis results had a high risk of bias, and the most common source of bias was that there was high heterogeneity between studies, and the sensitivity can only be reduced by a random effect model, and some studies (four items) did not clearly describe the confounders that they controlled. CONCLUSION: The limited evidence obtained in this study proves that the new puncture method does not have more advantages than the traditional technique, so it is no longer meaningful to continue to obsess over the impact of the puncture method on surgical outcome.

Laparoscopic versus robot-assisted pyeloplasty in infants and young children.

OBJECTIVE: To compare the characteristics of conventional laparoscopic pyeloplasty (LP) and robotic-assisted laparoscopic pyeloplasty (RALP) in infants and young children with ureteropelvic junction obstruction (UPJO). METHODS: We performed a retrospective study of patients (age: 0-36 months) who underwent dismembered pyeloplasty (Anderson-Hynes) with the fourth-generation RALP or traditional LP between April 2020 and December 2020. RESULTS: A total of 33 patients with UPJO were enrolled: 12 underwent RALP (9 left side; 3 right side) and 21 underwent LP (18 left side; 3 right side). In the RALP group, the median patient age was 17 months (range: 5-36 months). In the LP group, the median patient age was 9 months (range: 2-36 months) (P = 0.182). The mean operation times were 120.25 ± 37.54 min (RALP) and 156.10 ± 51.11 min (LP) (P = 0.042), and the mean lengths of hospital stay were 6.42 ± 1.62 days (RALP) and 8.19 ± 2.25 days (LP) (P = 0.023). Removal of the drainage tube was performed after 3.08 ± 0.69 days (RALP) and after 4.76 ± 1.81 days (LP) (P = 0.001). The postoperative pain showed no significant difference. The mean hospitalization costs were 61464.75 ± 2800.53 yuan (RALP) and 22169.52 ± 3442.15 yuan (LP) (P < 0.001). The mean follow-up time was 10-18 months. Significant improvements in the anteroposterior diameter and parenchymal thickness were observed after surgery. Conversion to laparotomy was not performed. No short-term complications occurred during postoperative hospitalization and follow-up. CONCLUSION: RALP has the advantages of less trauma and faster recovery. It can be safely and effectively performed in infants and young children, and its effectiveness is similar to that of traditional LP.

The role of neuroimmune and inflammation in pediatric uremia-induced neuropathy.

Uremic neuropathy in children encompasses a wide range of central nervous system (CNS), peripheral nervous system (PNS), autonomic nervous system (ANS), and psychological abnormalities, which is associated with progressive renal dysfunction. Clinically, the diagnosis of uremic neuropathy in children is often made retrospectively when symptoms improve after dialysis or transplantation, due to there is no defining signs or laboratory and imaging findings. These neurological disorders consequently result in increased morbidity and mortality among children population, making uremia an urgent public health problem worldwide. In this review, we discuss the epidemiology, potential mechanisms, possible treatments, and the shortcomings of current research of uremic neuropathy in children. Mechanistically, the uremic neuropathy may be caused by retention of uremic solutes, increased oxidative stress, neurotransmitter imbalance, and disturbance of the blood-brain barrier (BBB). Neuroimmune, including the change of inflammatory factors and immune cells, may also play a crucial role in the progression of uremic neuropathy. Different from the invasive treatment of dialysis and kidney transplantation, intervention in neuroimmune and targeted anti-inflammatory therapy may provide a new insight for the treatment of uremia.

Individual treatment strategy for single urethrocutaneous fistula after hypospadias repair: a retrospective cohort study.

Background: Urethrocutaneous fistula (UCF) remains the most common complication after hypospadias repair, and the recurrence rate of UCF is still high if the surgical techniques is not chosen properly, which called for better approaches to this problem. UCF presents different clinical characteristics due to their different locations and sizes, therefore we retrospectively analyzed the effects of different surgical techniques on single UCF after hypospadias repair in order to reduce the recurrence rates of UCF. Methods: A total of 602 patients diagnosed with UCF after hypospadias repair from January 2014 to December 2021 were enrolled. Baseline clinical characteristics such as age of patients, UCF location, size, surgical techniques were recorded. Patients were followed up in the outpatient clinic. The recurrence of UCF was defined as outcomes. Patients were divided according to the location of the UCF into a coronal UCF group and a non-coronal UCF group, which was then further classified according to the diameter of the UCF. The surgical technique and the recurrence rate of different types of UCF were analyzed and summarized. Results: A total of 425 patients satisfied the inclusion criteria and 71 patients (16.7%) had recurrent UCF. Five surgical techniques were used to repair the UCF, namely tubularized incised plate (TIP) urethroplasty, Mathieu urethroplasty, double ligation, simple classical closure and trap-door procedure. The recurrence rate was 24.1%, 14.3%, 15.1%, 16.7%, and 22.2%, respectively. TIP or Mathieu urethroplasty is recommended for patients with coronal UCF with glans dehiscence or patients with coronal UCF diameter ≥4 mm without glans dehiscence. In patients with coronal UCF without glans dehiscence, double ligation is recommended for small UCF with diameter <2 mm, and simple classical closure is recommended for UCF with diameter 2-<4 mm. In patients with non-coronal UCF, double ligation is recommended for UCF with diameter <3 mm, and simple classical closure is recommended for UCF with diameter ≥3 mm. Conclusions: Single UCF can be classified according to the location and size of the UCF. Different types of UCF should be treated with more appropriate individualized strategies, which can effectively reduce the recurrence rate of UCF.

Novel androgen receptor gene variant containing a frameshift mutation in a patient with complete androgen insensitivity syndrome.

A variety of mutations in the androgen receptor (AR) gene are linked to androgen insensitivity syndrome (AIS). AIS is the most common specific cause of 46, XY disorder in sex development. Here, we reported a patient which presented as a female with 46, XY karyotype and normal female external genitalia. The patient was diagnosed with complete AIS caused by a novel mutation (NM_000044, c.2678-2726del, p. Pro893Leufs*35) in the AR gene. Targeted exome sequencing was used to detect the patient's androgen receptor gene mutations. Sanger sequencing was used to validate the mutation. This study showed that a novel mutation of the AR gene can cause complete AIS; the study also broadened the AR mutation spectrum and indicated that targeted exome sequencing could help facilitate the diagnosis of complicated disorders in sexual development.