Identification of a novel WAS mutation and the non-splicing effect of a second-site mutation in a Chinese pedigree with Wiskott-Aldrich syndrome.

BACKGROUND: Wiskott-Aldrich syndrome (WAS) is a rare X-linked immunodeficiency disorder caused by abnormal expression of the WAS protein (WASp) due to mutations in the WAS gene, and is generally characterized by microthrombocytopenia, eczema, recurrent infections, and high susceptibility to autoimmune complications and hematological malignancies. RESULTS: Herein, we identified a novel WAS mutation (c.158 T > C) using next-generation sequencing in a Chinese pedigree with WAS. The expression of WASp in the patients and their families was detected by flow cytometry and western blot analysis. To explore the exon-splicing effect of intron mutations and the correlation between the genotype and clinical phenotype, four groups of wild-type (WT), exon mutant, intron mutant, and combined mutant recombinant plasmids were transfected into COS-7 cells in vitro. The proband showed dramatically decreased WASp expression, while the female carriers showed a slightly lower level of WASp. The expression of products in the mutant and WT recombinant plasmids was detected by real-time fluorescence quantitative polymerase chain reaction (PCR), which showed a significant reduction in the combined mutant group than in the WT, exon mutant, and intron mutant groups. The length of the expression products in the four groups showed no differences, each containing 360 base pairs. Sequence analysis confirmed that the c.158 T > C mutation appeared in the exon mutant and combined mutant groups, whereas the intron variant c.273 + 14C > T caused no other sequence changes. CONCLUSION: This study confirmed that the intron mutation did not affect the splicing of exons and excluded the influence of the double mutations at the transcription level on the severe clinical manifestations in the cousin. This in vitro study provided new insights into the pathogenesis of intronic mutations in WAS.

Comparison of percutaneous transforaminal endoscopic discectomy and open lumbar discectomy for lumbar disc herniations: A systematic review and meta-analysis.

Purpose: In order to compare the outcomes of percutaneous transforaminal endoscopic discectomy (PTED) and open lumbar discectomy (OLD) for lumbar disc herniation (LDH). Methods: The Pubmed, Cochrane Library, Web of Sience, Embase, Clinicaltrials.gov, CBM, CNKI, VIP, Wangfang databases were searched from inception to April 30, 2022 to collect the published studies about PTED vs. OLD for treatment of LDH. The Revman 5.2 was used for data analysis. The primary outcomes were excellent rates, complication rates and reoperation rates. The secondary outcomes were length of incision, length of operation, length of hospital stay, and the amount of intraoperative blood loss. Results: A total of nine studies were included, of which, eight randomized controlled trials and one retrospective study involving 1,679 patients with LDH (755 patients for PTED, and 924 patients for OLD) were included. According to meta-analysis, there were no significant difference in excellent rates (odds ratio [OR] = 1.47, 95% confidence intervals [CI]: 0.94-2.28, P = 0.09), reoperation rates (OR = 0.96, 95% CI: 0.50-1.84, P = 0.90), length of operation [standardized mean differences (SMD) = -17.97, 95%CI: -54.83-18.89, P = 0.34], and the amount of intraoperative blood loss (SMD = -128.05, 95%CI: -258.67-2.57, P = 0.05), respectively. There were significant differences in complication rates (OR = 0.22, 95% CI: 0.14-0.33, P < 0.001), length of incision (SMD = -2.76, 95%CI: -2.88--2.65, P < 0.001), and length of hospital stay (SMD = -5.19, 95%CI: -5.36--5.01, P < 0.001), respectively. Conclusions: PTED can achieve better outcomes with respect to the complication rates, length of incision, and length of hospital stay compared with OLD.

Natural Polysaccharide ß-Glucan Protects against Doxorubicin-Induced Cardiotoxicity by Suppressing Oxidative Stress.

Doxorubicin (DOXO) can be used to treat a variety of human tumors, but its clinical application is limited due to severe cardiotoxic side effect. Here, we explore the role of ß-glucan in DOXO-induced cardiotoxicity in mice and study its underlying mechanism. When co-administered with DOXO, ß-glucan was observed to prevent left ventricular dilation and fibrosis. In fact, DOXO reduces the activity of mitochondrial respiratory chain complex and enhances oxidative stress, which in turn impairs heart function. DOXO decreases the ATP production capacity of the heart and increases the ROS content, while ß-glucan can restore the heart capacity and reduce oxidative stress. ß-glucan also increases the activity of antioxidant enzymes GSH-PX and SOD, and reduces the level of MDA in the serum. In addition, the mRNAs of cardiac dysfunction marker genes ANP, BNP and Myh7 were significantly increased after DOXO induction, however, they did not increase when combined with ß-glucan administration. In conclusion, our results indicate that ß-glucan can improve the antioxidant capacity of the heart, thereby serving as a potential therapeutic strategy to prevent DOXO-induced cardiotoxicity.

Pyroptosis inhibition alleviates potassium oxonate- and monosodium urate-induced gouty arthritis in mice.

OBJECTIVES: Pyroptosis has been found implicated in several diseases, however, whether it was involved in gouty arthritis remained unclear. Our study was performed to uncover the role of pyroptosis in gouty arthritis based on a mice model. METHODS: Mouse gouty arthritis model was established by injections of potassium oxonate (PO), monosodium urate (MSU) and pyroptosis suppressor disulfiram. The diameter of the ankle joints was measured, and ankle joints morphology was observed with hematoxylin-eosin (H&E) staining. Uric acid, creatinine and blood urea nitrogen (BUN) concentrations were measured, while cytokines level and xanthine oxidase (XOD) activity were quantified. Relative pyroptosis markers expressions were determined using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot as needed. RESULTS: In mouse model, PO and MSU injections cause damage to right ankle, increase the root thickness ratio and uric acid, creatinine and BUN levels in serum and decrease the uric acid and creatinine levels in urine. Also, under PO and MSU treatment, up-regulated XOD activity, inflammatory cytokines levels and pyroptosis markers expressions are observed. Negative regulation of mice injury by disulfiram treatment is also observed. CONCLUSION: Pyroptosis inhibition might alleviate PO- and MSU-induced gouty arthritis, providing possible therapeutic strategies for gouty arthritis.

Case Report: Wiskott-Aldrich Syndrome Caused by Extremely Skewed X-Chromosome Inactivation in a Chinese Girl.

Wiskott-Aldrich syndrome (WAS) is a rare X-linked immunodeficiency disorder caused by abnormal expression of Wiskott-Aldrich syndrome protein due to WAS gene mutation, which is generally characterized by microthrombocytopenia, eczema, recurrent infections, and high risk of autoimmune complications and hematological malignancies. Although affected males with WAS usually manifest severe symptoms, female carriers have no significant clinical manifestations. Here, we describe a Chinese girl diagnosed with WAS carrying a heterozygous missense mutation in exon 2 of the WAS gene. The patient presented with persistent thrombocytopenia with small platelets and decreased WAS protein detected by flow cytometry and western blot analysis. The methylation analysis of the HUMARA gene displayed an extremely skewed X-chromosome inactivation (SXCI) pattern, where the X-chromosomes bearing normal WAS gene were predominantly inactivated, leaving the mutant gene active. Hence, our results suggest that completely inactivating the unaffected paternal X-chromosomes may be the reason for such phenotype in this female patient. SXCI has important implications for genetic counseling of female carriers with a family history of WAS.

Arachidonic Acid Promotes Intestinal Regeneration by Activating WNT Signaling.

Intestinal regeneration is crucial for functional restoration after injury, and nutritional molecules can play an important role in this process. Here, we found that arachidonic acid (AA) serves as a direct proliferation promoter of intestinal epithelial cells that facilitates small intestinal regeneration in both three-dimensional cultured organoids and mouse models. As shown in the study, during post-irradiation regeneration, AA positively regulates intestinal epithelial cell proliferation by upregulating the expression of Ascl2 and activating WNT signaling, but negatively regulates intestinal epithelial cell differentiation. AA acts as a delicate regulator that efficiently facilitates epithelial tissue repair by activating radiation-resistant Msi1+ cells rather than Lgr5+ cells, which are extensively considered WNT-activated crypt base stem cells. Additionally, short-term AA treatment maintains optimal intestinal epithelial homeostasis under physiological conditions. As a result, AA treatment can be considered a potential therapy for irradiation injury repair and tissue regeneration.

Synergistic protective effect of FTY720 and vitamin E against simulated cerebral ischemia in vitro.

The purpose of the present study was to explore the combination effect of FTY720 and vitamin E on cerebral ischemia. Astrocytes were isolated from newborn Sprague­Dawley rats and were subjected to FTY720, vitamin E, or combination of the two. The astrocyte cultures were then exposed to oxygen­glucose deprivation (OGD) to simulate an ischemic model in vitro. Cell viability, lactate dehydrogenase (LDH) leakage and cell apoptosis were detected following 12 h of exposure to OGD. In addition, the levels of tumor necrosis factor (TNF)­α, interleukin (IL)­6, IL­1ß, total antioxidant capacity, intercellular adhesion molecule (ICAM)­1, vascular cell adhesion molecule (VCAM)­1, chemokine (C­X­C motif) ligand (CXCL)­10, heme oxygenase (HO)­1 and superoxide dismutase (SOD)­1 were measured. Pre­treatment with FTY720 or vitamin E significantly elevated the cell viability and decreased LDH release and number of apoptotic cells. Combination treatment with FTY720 and vitamin E demonstrated a synergistic protective effect on OGD­induced cell viability, toxicity and apoptosis. Pre­treatment with FTY720 markedly reduced the release of IL­1ß, TNF­α, IL­6, ICAM­1, VCAM­1 and CXCL­10, and pre­treatment with vitamin E increased the levels of antioxidant, HO­1 and SOD­1. However, pre­treatment with FTY720 combined with vitamin E revealed a synergistic effect. Pre­treatment with FTY720 combined with vitamin E exerts synergistic neuroprotective effects in the simulated cerebral ischemia in vitro.