Clinical and molecular characterization of 10 Chinese children with congenital adrenal hyperplasia due to 11beta-hydroxylase deficiency.

BACKGROUND: The clinical manifestations of nonclassical 11beta-hydroxylase deficiency are very similar to those of non-classical 21-hydroxylase deficiency. For this study, we investigated the relationship between the clinical and molecular features of congenital adrenal hyperplasia caused by 11beta-hydroxylase deficiency and reviewed the related literature, which are expected to provide assistance for the clinical diagnosis and analysis of congenital adrenal hyperplasia. METHODS: Clinical data for 10 Chinese patients diagnosed with congenital adrenal hyperplasia in our hospital from 2018 to 2022 were retrospectively analyzed. We examined the effects of gene mutations on protease activity and constructed three-dimensional structure prediction models of proteins. RESULTS: We describe 10 patients with 11beta-hydroxylase gene mutations (n = 5, 46,XY; n = 5, 46,XX), with 10 novel mutations were reported. Female patients received treatment at an early stage, with an average age of 2.08 ± 1.66 years, whereas male patients received treatment significantly later, at an average age of 9.77 ± 3.62 years. The most common CYP11B1 pathogenic variant in the Chinese population was found to be c.1360C > T. All mutations lead to spatial conformational changes that affect protein stability. CONCLUSIONS: Our study found that there was no significant correlation between each specific mutation and the severity of clinical manifestations. Different patients with the same gene pathogenic variant may have mild or severe clinical manifestations. The correlation between genotype and phenotype needs further study. Three-dimensional protein simulations may provide additional support for the physiopathological mechanism of genetic mutations.

Ubiquitylation of cyclin C by HACE1 regulates cisplatin-associated sensitivity in gastric cancer.

BACKGROUND: Cyclin C (CCNC) was reported to take part in regulating mitochondria-derived oxidative stress under cisplatin stimulation. However, its effect in gastric cancer is unknown. This study aimed to investigate the role of cyclin C and its ubiquitylation in regulating cisplatin resistance in gastric cancer. METHODS: The interaction between HECT domain and ankyrin repeat-containing E3 ubiquitin-protein ligase 1 (HACE1) and cyclin C was investigated by GST pull-down assay, co-immunoprecipitation and ubiquitylation assay. Mitochondria-derived oxidative stress was studied by MitoSOX Red assay, seahorse assay and mitochondrial membrane potential measurement. Cyclin C-associated cisplatin resistance was studied in vivo via xenograft. RESULTS: HACE1 catalysed the ubiquitylation of cyclin C by adding Lys11-linked ubiquitin chains when cyclin C translocates to cytoplasm induced by cisplatin treatment. The ubiquitin-modified cyclin C then anchor at mitochondira, which induced mitochondrial fission and ROS synthesis. Depleting CCNC or mutation on the ubiquitylation sites decreased mitochondrial ROS production and reduced cell apoptosis under cisplatin treatment. Xenograft study showed that disrupting cyclin C ubiquitylation by HACE1 conferred impairing cell apoptosis response upon cisplatin administration. CONCLUSIONS: Cyclin C is a newly identified substrate of HACE1 E3 ligase. HACE1-mediated ubiquitylation of cyclin C sheds light on a better understanding of cisplatin-associated resistance in gastric cancer patients. Ubiquitylation of cyclin C by HACE1 regulates cisplatin-associated sensitivity in gastric cancer. With cisplatin-induced nuclear-mitochondrial translocation of cyclin C, its ubiquitylation by HACE1 increased mitochondrial fission and mitochondrial-derived oxidative stress, leading to cell apoptosis.

Treatment with retinoic acid and lens epithelial cell-conditioned medium in vitro directed the differentiation of pluripotent stem cells towards corneal endothelial cell-like cells.

Embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) have extensive self-renewal capacity and the potential to differentiate into all tissue-specific cell lineages, including corneal endothelial cells (CECs). They are a promising prospect for the future of regenerative medicine. The method of derivation of CECs from ESCs and iPSCs, however, remains to be elucidated. In this study, mouse ESCs and iPSCs were induced to differentiate into CECs using CEC embryonic development events as a guide. All-trans retinoic acid (RA) treatment during the embryoid body (EB) differentiation step was used to promote neural crest (NC) cell differentiation as first step and was followed by a second induction in CEC- or lens epithelial cell (LEC)-conditioned medium (CM) to ultimately generate CEC-like cells. During the corresponding differentiation stages, NC developmental markers and CEC differentiation markers were detected at the protein level using immunocytochemistry (ICC) and at the mRNA level by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). During the first stage, the data indicated that 4 days of treatment with 1 µM RA starting on day 4 of EB formation favored NC cell differentiation and that plating on gelatin-coated plates led to cell migration out of the EBs. The second-stage differentiation results showed that the CM, particularly the LEC-CM, enhanced the yield of polygonal cells with CEC-specific marker expression shown by ICC and RT-qPCR. This study demonstrates that mouse ESCs and iPSCs were induced and expressed CEC differentiation markers when subjected to a two-step inducement process, suggesting that they are a promising resource for corneal endothelium failure replacement therapy in the future.

[Bulbourethral sling for treating urinary incontinence due to intrinsic sphincter deficiency].

OBJECTIVE: To investigate the therapeutic effect of bulbourethral sling for treating urinary incontinence resulted from intrinsic sphincter deficiency. METHODS: Eighteen male patients with urinary incontinence were treated by bulbourethral sling operation. RESULT: In the follow-up for 1 to 34 months, 16 patients achieved clinical cure and 2 showed significant improvement of the symptoms. CONCLUSION: This operation can be effective for treating urinary incontinence resulted from intrinsic sphincter deficiency.

[Laparoendoscopic single-site surgery for male pseudohermaphroditism: the initial report].

OBJECTIVE: To evaluate the therapeutic effect of laparoendoscopic single-site surgery (LESS) for treatment of male pseudohermaphroditism. METHODS: A 17-year-old patient with male pseudohermaphroditism and a female social sex was admitted. According to the request by the patient and the relatives for a female gender, LESS vaginoplasty and cryptorchidectomy were performed using a single multilumen port inserted through a 2.5 cm incision below the umbilicus, followed by reconstruction of the perineal region by open surgery. RESULTS: The total operative time was 7 h, and the LESS procedure lasted for about 3.5 h. No other port incision was needed. The estimated intraoperative blood loss was 400 ml. No electrolyte or metabolic acid-base balance disorders were observed perioperatively. In the follow-up examination at 6 months after the operation, the reconstructed vagina healed smoothly without obvious contraction or fixation failure, and the perineal region showed good appearance. CONCLUSION: With minimal invasiveness, LESS surgery produces good cosmetic effect and allows rapid postoperative recovery, thus may become a promising alternative to the management of pseudohermaphroditism.

[NSP5/NSP6 of rotavirus strain TB-Chen and preliminary genotyping analysis].

Strain TB-Chen is a group A rotavirus (RV) isolated from a Chinese infant suffering from gastroenteritis in hospital. The NSP5 and NSP6 of strain TB-Chen are encoded by the 10th gene segment (816bp in whole length) of the viral genome. The results obtained in this study showed that the NSP5 was encoded in the first open-reading-frame (ORF) of the gene segment (from 22bp to 624bp), and NSP6 was encoded in the second ORF (from 80bp to 355bp). The NSP5 protein consisted of 200 amino acid residues with a putative molecular mass of 21.9 kD, and a putative isoelectric point of 7.86. The NSP6 protein consisted of 92 amino acids with a putative molecular mass of 11 kD, and a putative isoelectric point of 9.65. This study further analyzed phylogenetic relationship of the NSP5/NSP6 ORF nucleotide sequence. The results showed that the NSP5s of group A rotavirus could be at least classified into 7 genotypes (H1-H7), the NSP6s could be at least classified into 8 genotypes (hl-h8); the genotypes of the NSP5 and NSP6 derived from strain TB-Chen was classified as H2 and h2. This was the first report on the genotype classification of the NSP6 of group A RVs, and it was proposed English letter "h" to represent genotype of the NSP6, e. g. strain 69M classified as H7h7, strains Wa and KU classified as H1h8.