Maternal Administration of Busulfan before in Utero Hematopoietic Cell Transplantation Improves Congenic Bone Marrow Cell Engraftment in a Murine Model.

In utero hematopoietic cell transplantation (IUHCT) is a nonmyeloablative procedure that leads to donor cell chimerism and donor-specific tolerance. However, most clinical applications of IUHCT have failed because of low levels or even no engraftment of donor cells in immunologically normal fetuses. It is likely that the competition from the host hematopoietic compartment is the primary barrier to successful IUHCT, suggesting that conditioning methods that provide a competitive advantage to donor cells may lead to higher-level engraftment following IUHCT. This study aimed to research whether maternal administration of low-dose total body irradiation (TBI) or busulfan (BU) before IUHCT may result in increased donor cell chimerism in postnatal bone marrow transplantation in a congenic murine model. We first determined the birth and mortality rates after maternal administration of low-dose TBI (0, 2 or 4 Gy) or BU (5, 10, 15, or 20 mg/kg) before IUHCT in B6 mice. The mice that received 2 Gy TBI plus IUHCT showed significantly lower birth rate (23.3%) and a 100% 3-day mortality rate. The mice that received 10 mg/kg BU plus IUHCT had similar birth and 3-day mortality rates (58.6% and 0%) compared to mice that received IUHCT alone (61.1% and 4.55%). We then performed maternal administration of BU at 1 of 3 dosages (5, 10, or 15 mg/kg) at 24 hours before intrauterine transplantation of 2.5 × 105 B6GFP Sca-1+ bone marrow cells (BMCs) or 2.5 × 106 B6GFP BMCs on gestational day 14 (E14). Green fluorescent protein (GFP) chimerism in peripheral blood mononuclear cells (PBMCs), RBCs, and platelets of mice at 4 weeks of age was enhanced significantly with an increase in BU dose. Moreover, GFP chimerism of PBMCs from the B6GFP BMC group was significantly higher than that of the B6GFP Sca-1+ BMC group (22.56% versus 7.20%; P = .018). Finally, the pregnant mice were treated with 10 mg/kg of BU at E13, E14, or E15, followed by intrauterine transplantation of 2.5 × 106 B6GFP BMCs 24 hours later. Except for the short-term level of chimerism in PBMCs, which showed no significant difference among the 3 study groups, the results indicate that both short-term (age 4 weeks) and long-term (age 14 weeks) engraftment in PBMCs, RBCs, and platelets was higher in group E16 compared with groups E14 and E15. We also discovered that the engraftment was stable, multilineage, and increased with time. In conclusion, maternal administration of BU, but not of TBI, along with IUHCT could significantly enhance engraftment in a congenic murine model.

[Curative effect analysis of radical endoscopic sinus surgery for eosinophilic chronic rhinosinusitis with nasal polyps].

Objective:To evaluate the efficacy of functional endoscopic sinus surgery（FESS） and radical endoscopic sinus surgery（RESS） in eosinophilic chronic sinusitis with nasal polyps（EosCRSwNP）. Methods:A total of 44 patients diagnosed with EosCRSwNP in the Department of Otorhinolaryngology Head and Neck Surgery, Henan Provincial People's Hospital from July 1st, 2020 to August 1st, 2021 were included, the percentage of eosinophils in leukocytes in all patients included was more than 3.05%. The patients were randomly divided into FESS group and RESS group according to random number table. The visual analogue scale （VAS） score, Lund-Kennedy score and sino-nasal outcome test-22 （SNOT-22） were compared between the two groups before operation, 1 month, 3 months, 6 months and 1 year after operation. Results:At 1 year after operation, the scores of the two groups were significantly improved compared with those before operation, and the differences were statistically significant （P<0.01）. There were significant differences in nasal endoscopic score, VAS score and SNOT-22 score between the two groups（P=0.01, P=0.03, P=0.03）. The recurrence rate of RESS group was 26.09%（6/23） and that of FESS group was 61.90%（13/21）, and the difference was statistically significant（P=0.04）. Conclusion:Both RESS and FESS can improve nasal symptoms and promote olfactory recovery in EosCRSwNP patients, but RESS has more advantages in reducing recurrence and improving the prognosis of patients.

Long noncoding RNA Gm44593 attenuates oxidative stress from age-related hearing loss by regulating miR-29b/WNK1.

Long noncoding RNA has been reported to play important role in various disease. However, the function of lncRNA in age-related hearing loss still unclear. The aim of our study is to investigate the function and mechanism of lncRNA Gm44593 in AHL. ATP content, JC-1 assay, mitochondrial content, cell death rates and dual-luciferase reporter assay were performed to assess the function of lncRNA Gm44593 in HEI-OC1 cells. The expression of lncRNA Gm44593 was significantly upregulated upon H2O2 and starvation treatment. Overexpression of lncRNA Gm44593 manifestly reduced the cell death rates. The ATP content, mtDNA content and mitochondrial membrane potential were alleviated upon overexpression of lncRNA Gm44593. We also proved that miR-29b is the direct target of lncRNA Gm44593. Overexpression of miR-29b completely restored the effect induced by lncRNA Gm44593. In addition, we provided evidences that WNK1 is the direct target of miR-29b. Our research uncovers a potential role of lncRNA Gm44593 in age-related hearing loss. We provide new insights into potential therapeutic targets for the amelioration of age-related hearing loss.

Hsa_circ_0006232 promotes laryngeal squamous cell cancer progression through FUS-mediated EZH2 stabilization.

Laryngeal squamous cell cancer (LSCC) is one of the most common malignant tumors in head and neck tumors. Our previous study has revealed that hsa_circ_0006232 is abnormally expressed in LSCC. This study attempts to verify the biological role of hsa_circ_0006232 in LSCC. We found that compared with human bronchial epithelial cells, hsa_circ_0006232 was highly expressed in human LSCC cells (AMC-HN-8 and TU686). Moreover, hsa_circ_0006232 promoted proliferation, migration and invasion of AMC-HN-8 and TU686 cells. Hsa_circ_0006232 promoted the expression of enhancer of zeste homolog 2 (EZH2) and repressed the expression of phosphatase and tensin homolog deleted on chromosome 10 (PTEN). Fused in sarcoma (FUS) interacted with hsa_circ_0006232 and EZH2, and FUS promoted the stabilization of EZH2. Hsa_circ_0006232 inhibited PTEN by promoting FUS expression. Moreover, we constructed a tumor xenograft model by injection of AMC-HN-8 cells with hsa_circ_0006232 knockdown, and we found that hsa_circ_0006232 deficiency decreased tumor growth in mice. Hsa_circ_0006232 silencing repressed EZH2 expression and enhanced PTEN expression in tumor tissues. In conclusion, our data have demonstrated that Hsa_circ_0006232 promotes proliferation, migration and invasion of LSCC cells, and accelerates tumor growth of LSCC through FUS-mediated EZH2 stabilization. Thus, hsa_circ_0006232 may be a novel therapeutic target in LSCC treatment.

Hsa_circ_0042823 accelerates cancer progression via miR-877-5p/FOXM1 axis in laryngeal squamous cell carcinoma.

BACKGROUND: Laryngeal squamous cell carcinoma (LSCC) is a common malignant tumour of the head and neck. Our previous study reveals that the circular RNA (circRNA) hsa_circ_0042823 is abnormally expressed in LSCC, suggesting that hsa_circ_0042823 is closely associated with LSCC. Here, we attempted to explore the molecular mechanism of hsa_circ_0042823 in LSCC. METHODS: Quantitative real-time PCR and western blot were performed to assess the expression of gene and protein in human laryngeal carcinoma cells, TU212 and TU686. MTT and transwell assays were performed to examine cell proliferation, migration and invasion. The relationship among hsa_circ_0042823, miR-877-5p and forkhead box M1 (FOXM1) was verified by luciferase reporter assay. Finally, we constructed a subcutaneous tumour mouse model to analyse in vivo growth of LSCC cells following knockdown of hsa_circ_0042823. RESULTS: Compared with normal human bronchial epithelial cells (HBECs), hsa_circ_0042823 was highly expressed in the LSCC cell lines (AMC-HN-8 and TU686). Further studies demonstrated that hsa_circ_0042823 interacted with miR-877-5p, and FOXM1 was the target of miR-877-5p. Hsa_circ_0042823 promoted the expression of FOXM1 via its ceRNA activity on miR-877-5p. Hsa_circ_0042823 overexpression promoted proliferation, migration and invasion of AMC-HN-8 cells through regulating miR-877-5p/FOXM1 axis. Additionally, inhibition of hsa_circ_0042823 inhibited growth of LSCC in vivo via miR-877-5p/FOXM1 axis. CONCLUSIONS: Hsa_circ_0042823/miR-877-5p/FOXM1 axis participates in the progression of LSCC. This work demonstrates that hsa_circ_0042823 accelerates cancer progression by regulating miR-877-5p/FOXM1 axis in LSCC. Therefore, this study may provide new insights into the pathogenesis of LSCC.KEY MESSAGESHsa_circ_0042823 promotes FOXM1 expression by sponging miR-877-5p.Hsa_circ_0042823 promotes proliferation, migration, invasion of LSCC cells.Hsa_circ_0042823 knockdown inhibits tumour growth of LSCC via miR-877-5p/FOXM1 axis.

miR-125a suppresses viability and glycolysis and induces apoptosis by targeting Hexokinase 2 in laryngeal squamous cell carcinoma.

BACKGROUND: miR-125a usually functions as a tumor suppressor in cancers. However, the role of miR-125a in laryngeal squamous cell carcinoma (LSCC) has not been determined. METHODS: qRT-PCR was applied to measure the expression of miR-125a and HK2 mRNA in LSCC tissues and cells. CCK-8 kit and flow cytometry analysis were performed to detect cell viability and apoptosis. Luciferase reporter assay and RNA immunoprecipitation (RIP) were conducted to confirm the relationship between miR-125a and HK2. Commercial test kits were used to determine the concentrations of glucose and l-lactate. Xenograft in mice was constructed to validate the function and mechanism of miR-125a in LSCC tumor growth. RESULTS: A negative correlation was found between miR-125a expression and the level of Hexokinase 2 (HK2) mRNA in LSCC tissues. Functional experiments found that miR-125a inhibited viability and glycolysis and induced apoptosis in LSCC cells. Similarly, HK2 downregulation led to viability and glycolysis inhibition and induction of apoptosis in LSCC cells in vitro. Moreover, miR-125a overexpression suppressed LSCC xenograft growth in vivo. Mechanically, HK2 was verified to be a target of miR-125a by luciferase reporter assays and RNA immunoprecipitation (RIP) assays. Furthermore, restored HK2 expression reversed miR-125a-mediated proliferation and glycolysis inhibition and induction of apoptosis in LSCC cells. CONCLUSIONS: miR-125a suppressed LSCC progression by targeting HK2 in vitro and in vivo, suggesting that miR-125a may be a potential molecular target for LSCC treatment.

A probable case of Legg-Calvé-Perthes disease in Warring States-era China.

Reports of Legg-Calvé-Perthes disease (LCPD) in the paleopathological literature are rare. Here, the authors present a probable case of LCPD, which presents as abnormal morphology of the proximal femur. The condition was observed in an individual of the Warring States period in Shaanxi Province, China, and the morphology involves a "mushroom head" deformity of the proximal right femur and an enlarged acetabulum, along with a contralateral tibia, talus, and navicular that are enlarged and demonstrate periosteal new bone formation. The authors consider tuberculosis, septic arthritis, trauma, slipped capital femoral epiphysis, and Legg-Calvé-Perthes disease in a differential diagnosis. The authors conclude that the most likely diagnosis for the deformity is Legg-Calvé-Perthes disease. Bony changes in the hip joint and contralateral lower leg suggest that the individual had an altered gait because of the condition.

[Expression of angiopoietin-1,2 and its relationship with angiogenesis in laryngeal squamous cell carcinoma].

OBJECTIVE: To investigate the expression of Angiopoietin-1,2 in laryngeal squamous cell carcinoma (LSCC) and the relationship between Angiopoietin-1,2 expression and clinicopathologic parameters and microvessel density (MVD) marked by CD105, and also evaluate the significance of co-expression of Ang-2 and vascular endothelial cell growth factor (VEGF) in tumor angiogenesis. METHOD: The expression of Ang-1,2 and VEGF in samples of tumor, para cancer and normal mucosa were detected by immunohistochemical method. RESULT: The expression of Angiopoietin-1,2 were identified in LSCC, para cancer tissue and normal mucosa. The VEGF expression was only existed in LSCC. The expression of Ang-1,2 were significantly higher in LSCC than in para cancer tissue (P < 0.05) and normal mucosa (P < 0.01). In the clinicopathologic parameters, only the expression of Ang-2 was closely correlated with clinical stages and MVD (all P < 0.05). There was no significant correlation between the expression of Ang-1,2 and tumor differentiation degree (all P > 0.05). When the expression of Ang-2 and VEGF were both positive, the mean value of MVD was higher than others (P < 0.05). CONCLUSION: These results suggest that overexpression of Ang-1,2 may play a very important role in the development of LSCC and are closely correlated with angiogenesis. Ang-2 promote angiogenesis when interacting with VEGF.