HuR deficiency abrogated the enhanced NLRP3 signaling in experimental ischemic stroke.

Human antigen R (HuR) is a universally expressed RNA-binding protein that plays an essential role in governing the fate of mRNA transcripts. Accumulating evidence indicated that HuR is involved in the development and functions of several cell types. However, its role in cerebral ischemia/reperfusion injury (CIRI) remains unclear. In this study, we found that HuR was significantly upregulated after CIRI. Moreover, we found that silencing HuR could inhibit the inflammatory response of microglia and reduce the damage to neurons caused by oxygen-glucose deprivation/reperfusion treatment. In vivo, we found that microglial HuR deficiency significantly ameliorated CIRI and reduced NLRP3-mediated inflammasome activation. Mechanistically, we found that HuR could regulate NLRP3 mRNA stability by binding to the AU-rich element (ARE) region within the 3' untranslated region (UTR) of NLRP3 mRNA. In addition, we found that the upregulation of HuR was dependent on the upregulation of NADPH oxidase-mediated ROS accumulation. Collectively, our studies revealed that HuR could regulate NLRP3 expression and that HuR deficiency abrogated the enhanced NLRP3 signaling in experimental ischemic stroke. Targeting HuR may be a novel therapeutic strategy for cerebral ischemic stroke treatment.

CST3 alleviates retinal vascular leakage by regulating the Rap1 signaling pathway.

Retinal vascular leakage is a major event in several retinal diseases, including diabetic retinopathy (DR). In a previous study, we demonstrated that the aqueous humor concentration of Cystatin C (CST3), a physiological inhibitor of cysteine protease, is negatively correlated with the severity of diabetic macular edema. However, its function in the retina has not been clearly elucidated. In this study, we found a significant decrease in the aqueous humor concentration of CST3 with DR progression. Furthermore, we found that CST3 was expressed in retinal endothelial cells and that its expression was significantly downregulated in high glucose-treated human retinal microvascular endothelial cells (HRMECs) and the retinal vessels of oxygen-induced retinopathy (OIR) mice. Silencing CST3 expression resulted in decreased HRMEC migration and tubule formation ability. Exogenous addition of the CST3 protein significantly improved HRMEC migration and tubular formation. In-vivo experiments demonstrated that CST3 silencing induced retinal vascular leakage in WT mice, while its intravitreal injection significantly reduced retinal leakage in OIR mice. Mechanistically, CST3 promoted the expression of the downstream adhesion molecules, claudin5, VE-cadherin, and ZO-1, in retinal vascular cells by regulating the Rap1 signaling pathway. Therefore, this study revealed a novel mechanism by which CST3 improves retinal vascular function and provided evidence that it is a potential therapeutic target for retinal vascular leakage.

Clinical Manifestations and Genetic Analysis of 17 Patients with Autosomal Dominant Hyper-IgE Syndrome in Mainland China: New Reports and a Literature Review.

PURPOSE: Autosomal dominant hyper-IgE syndrome (AD-HIES) is a rare complicated primary immunodeficiency disease (PID). Signal transducer and activator of transcription 3 (STAT3) gene mutation is found to cause AD-HIES. The distribution of AD-HIES patients with STAT3 deficiency in the Chinese population is not clear. Herein, we retrospectively report 17 AD-HIES patients with STAT3 deficiency and demonstrate their clinical, immunological, and genetic features. METHODS: Patients' clinical data were collected from their medical records. Routine laboratory testing results included lymphocyte subset analysis and immunoglobulin quantification. STAT3 mutations were investigated by sequencing of genomic DNA. RESULTS: Among 575 patients with PID, 28 (4.87%) were clinically diagnosed as HIES. Among them, 17 (2.96%) were confirmed as STAT3 mutant AD-HIES. The ratio of male to female patients was 8:9. All of the 17 patients had NIH scores over 40 points. The mean ages at onset and diagnosis were 1.05 and 10.35 years, respectively. Three patients (17.65%, 3/17) died with a mean age of 13.33 years. Eczema, recurrent skin infection, and respiratory tract infection were the most common clinical symptoms and are present in all of the 17 patients in this study. Six patients (37.5%, 6/16) suffered complication from BCG vaccination. Noninfection symptoms are characteristic facial features in 17 patients (100%, 17/17), retention of primary teeth in 10 patients (90.91%, 10/11), and abnormal bone fractures in 7 patients (41.18%, 7/17). Eleven types of STAT3 mutations were identified in 17 patients, including 1 novel mutation. CONCLUSIONS: We here retrospectively report the largest Chinese cohort of AD-HIES patients with STAT3 mutation. Unique features, when compared to existing literature reports, include (1) later age of diagnosis, (2) significantly higher rate of BCG complications, and (3) lower rate of candidiasis and chronic otitis media.

Genome-Wide ChIP-seq and RNA-seq Analyses of STAT3 Target Genes in TLRs Activated Human Peripheral Blood B Cells.

Toll like receptors (TLRs) induced response plays a vital role in B-cell development and activation, in which TLR7-mediated and TLR9-mediated response interact together and play antagonistic or cooperative roles at different situations. Previous studies showed that the transcription factor signal transducer and activator of transcription (STAT) 3 was one of the key transcriptional factors (TFs) needed for both TLR7 and TLR9 signaling in B cell, and patients with autosomal dominant hyper IgE syndromes (AD-HIES) due to STAT3 mutations having defective TLRs response in B cells. However, how STAT3 affects its target genes and the downstream signaling pathways in B cell upon TLRs stimulation remains unclarified on a genome-wide level. ChIP-seq and RNA-seq was used in this study to identify the STAT3 targets in response to TLRs stimulation in human B cell. STAT3 ChIP-seq results showed a total of 611 and 2,289 differential STAT3-binding sites in human B cell after TLR7 and TLR9 agonists stimulation, respectively. RNA-seq results showed 1,186 and 1,775 differentially expressed genes after TLR7 and TLR9 activation, respectively. We identified 47 primary STAT3 target genes after TLR7 activation and 189 target genes after TLR9 activation in B cell by integration of STAT3 ChIP-seq and RNA-seq data. Among these STAT3 primary targets, we identified 7 TFs and 18 TFs for TLR7 and TLR9 response, respectively. Besides, we showed that STAT3 might regulate TLR9, but not TLR7 response in B cells through directly regulating integrin signaling pathway, which might further affect the antagonism between TLR7 and TLR9 signaling in B cell. Our study provides insights into the molecular mechanism of human TLRs response in B cell and how it can be regulated, which helps to better understand and modulate TLR-mediated pathogenic immune responses in B cell.

Eustachian tube teratoma: A case report.

BACKGROUND: Mature teratoma composed of all three basic germ cell layers of the head and neck is a rare disease. Teratomas involving the temporal bone are particularly scarce. CASE SUMMARY: A 48-year-old male patient with a history of chronic otitis of the left ear from infancy, for which he had been operated on twice, was referred to our hospital for chronic otitis, cholesteatoma and a middle ear mass. Computed tomography (CT) scan and magnetic resonance imaging (MRI) revealed a eustachian tube teratoma, in which the anterior lower part and posterior upper part were connected by a thin membranaceous tissue. The mass was removed completely under general anesthesia by mastoidectomy. As of last follow-up (2 years post-surgery), the disease had not relapsed. CONCLUSION: Pre-operative CT and MRI are necessary for eustachian tube teratoma. Complete surgical resection provided excellent prognosis.

Hepatic steatosis with mass effect: A case report.

BACKGROUND: Hepatic steatosis is a common radiologic finding. Some imaging inklings are the absence of a mass effect, and there is currently no report of hepatic steatosis with mass effect. CASE SUMMARY: A 23-year-old female was admitted due to a liver mass for half a month. No obvious abnormalities were found in physical and laboratory examinations. Ultrasound, computed tomography, and magnetic resonance imaging showed a huge mass between the liver and stomach with a significant mass effect, and the caudate lobe and left lobe of the liver were involved. The signal on T2- and T1- weighted fat-saturated images of the mass was significantly reduced, and the enhanced scan showed inhomogeneous enhancement. Surgical and pathological findings indicated the diagnosis of hepatic steatosis. The operation and re-review of the patient's images showed that the lesion was supplied by the branch of the hepatic artery. The signal on T1-weighted out-of-phase images of the lesion was lower than on in-phase images, and there was no black rim cancellation artifact around the hepatic steatosis area on T1-weighted out-of-phase images. The dynamic enhancement pattern of the lesion was similar to that of the adjacent normal liver parenchyma. The above characteristics suggested that the lesion was hepatic steatosis. However, in this case, the lesion showed exogenous growth and was mass-like, with an obvious mass effect, which has not been reported previously. CONCLUSION: Hepatic steatosis could grow exogenously and has an obvious mass effect. It needs to be distinguished from fat-rich tumors. The T1-weighted in- and out-of-phase images and dynamic enhanced scanning are valuable for differential diagnosis of this lesion.

MTDH antisense oligonucleotides reshape the immunosuppressive tumor microenvironment to sensitize Hepatocellular Carcinoma to immune checkpoint blockade therapy.

Immune checkpoint blockade (ICB) therapy is an important treatment option for individuals with cancer, but it has certain limitations. Identifying a better target that can overcome tumor immune escape and stimulate T cell activity is critical. This research aimed to delve into the molecular mechanism underlying the immunoregulatory function of metadherin (MTDH), which is a novel and potential therapeutic target in hepatocellular cancer (HCC). A small interfering RNA library was screened using the luciferase reporter assay and PD-L1 promoter. The Cancer Genome Atlas database and HCC tissues were used to investigate the relationship between MTDH and PD-L1. The association between MTDH and ß-catenin/lymphoid enhancer binding factor (LEF-1) was discovered by co-immunoprecipitation. The chromatin immunoprecipitation assay was used to investigate the interaction of MTDH with the PD-L1 promoter when LEF-1 expression was silenced. Locked nucleic acid antisense oligonucleotides (ASOs) were used to inhibit MTDH. We utilized in vitro co-cultures and in vivo syngeneic tumor development experiments to confirm the effectiveness of MTDH ASO combined with PD-1 monoclonal antibody (mAb). MTDH was demonstrated to be a PD-L1 modulator. MTDH increased PD-L1 expression and upregulated PD-L1 transcriptional activity through ß-catenin/LEF-1 signaling. More importantly, MTDH ASO improved the anti-PD-1 response and increased cytotoxic T-cell infiltration in PD-1 mAb-treated malignancies. MTDH effectively predicts the therapeutic efficacy of ICB therapy. Our results imply that combining MTDH ASO with PD-1 mAb could be a promising therapeutic strategy for HCC. In addition, MTDH is a potential novel biomarker for predicting the effectiveness of immune checkpoint inhibitor treatment.

Immunoregulatory effect of human ß-defensin 1 on neonatal cord blood monocyte-derived dendritic cells and T cells.

The relationship between breastfeeding and infant health has been well elucidated in past decades. Our previous study has shown that human ß-defensin 1 (hBD-1) in human breast milk plays a protective role in reducing the incidence of upper respiratory infection in infants younger than 6 months. In the present study, we aim to reveal the mechanism underlying the protective role of hBD-1 by focusing on its immunoregulatory function in neonates. Cord blood (CB) from newborns' umbilical cords, which can simulate many of the neonatal symptoms, was used to study the immunomodulatory role of hBD-1 in neonates in vitro. Our results showed that hBD-1 promotes the GM-CSF- and IL-4-driven differentiation of neonatal umbilical CB monocytes to immature dendritic cells (DCs) and the final maturation of CB monocyte-derived DCs (moDCs) induced by LPS but not inflammatory cytokine production. In addition, hBD-1 inhibits apoptosis in neonatal moDCs through CCR6, which might be a possible mechanism of the hBD-1-induced phenotypes in moDCs. Furthermore, we found that hBD-1 promotes the proliferation and activation, but not the maturation, of neonatal CB CD4 + T cells. These results extend the immunoregulatory effects of hBD-1 and provide a potential mechanism for the protective role of hBD-1 in early infants, which will inform the development of infant nutrition, novel vaccines and anti-infective strategies in the future.

Clinical, Laboratory, and Molecular Characteristics and Remission Status in Children With Severe Congenital and Non-congenital Neutropenia.

Objectives: Severe congenital neutropenia (SCN) is a primary immunodeficiency disease characterized by the early onset of recurrent infections and persistent severe neutropenia, with or without genetic defect. We aimed to study the different clinical features and hematological and bone marrow characteristics of patients with SCN and the non-congenital form of severe neutropenia (SN) with unknown etiology. Methods: Thirty-nine Chinese children with severe neutropenia for longer than 6 months unrelated to virus infection or autoimmune diseases were enrolled in the study to analyse the clinical, laboratory, and molecular characteristics. They were followed clinically to observe their remission status. Results: Seven patients were found to have SCN mutations, including ELANE and G6PC3. Among 26 patients with close follow-up, one died for an unknown reason, and 10 resolved spontaneously with a median neutropenia duration of 14.5 months; these patients were designated as having recovered SN. The demographic characteristics of both groups were similar, with a median infection rate of 5 times/year. SCN patients had more frequent infection than recovered SN patients (4 times/year, P = 0.039). The median absolute neutrophil count (ANC) was 0.40 × 109/L in SCN patients, which was significantly higher than 0.2 × 109/L in SN with unknown etiology and 0.21 × 109/L in recovered SN patients (P = 0.021, P = 0.017). The median monocyte count was 1.60 × 109/L in SCN patients, which was also significantly higher than 0.57 × 109/L in SN of unknown etiology and 0.55 × 109/L in recovered SN patients (P = 0.018, P = 0.001). Bone marrow examinations demonstrated myeloid maturation arrest at the myelocyte-metamyelocyte stage in SCN patients and normal findings in SN with unknown etiology and recovered SN patients. Conclusions: Patients with severe neutropenia due to gene mutations demonstrate more serious symptoms than patients with unknown etiology. Patients with relatively higher ANC and monocyte counts are more likely to have known gene mutations. Future studies should focus on more detailed laboratory investigation, prolonged follow-up and advanced molecular biology tools to facilitate accurate diagnosis and effective treatment.

HRCT and MRI findings in X-linked non-syndromic deafness patients with a POU3F4 mutation.

OBJECTIVE: The aim of this study was to analyze HRCT and MRI findings in patients with X-linked non-syndromic deafness and a POU3f4 mutation. METHODS: HRCT and MRI data of four patients (males, 2-19 years old) with a POU3f4 mutation were collected and a retrospective review was performed. Cochlea, internal auditory canal (IAC), vestibule, semicircular canals, vestibular aqueduct, nerve canals in the IAC fundus, stapes and cochlear nerve were evaluated on 2D images (multi-planner reformation, MPR) and cochlear foramen on 3D images (CT virtual endoscopy, CTVE). Ten cases with normal hearing subjected to CT and MR exams served as controls. RESULTS: Inner ear malformations were bilateral and symmetrical. Cochlear malformation was shown to consist of as a relatively normal outer coat shape, absence of a cochlear modilous, and a direct intercommunication between the IAC and cochlear inner cavity. The lateral portion of the IAC was dilated. A spiral cochlear inner cavity was observed with CTVE images versus a helical cochlear nerve foramen as seen in controls. The labyrinthine facial nerve canal and superior vestibular nerve canal were enlarged. The Bill's bar was hypertrophic and partially pneumatized. A thickened stapes footplate was present and a fissura ante fenestram was absent in seven ears examined. A column shaped stapes was observed in one ear. CONCLUSIONS: The absence of a cochlear modilous with a dilated lateral IAC and thickened stapes footplate were the remarkable features observed with imaging these in X-linked non-syndromic deafness patients with a POU3F4 mutation. Preoperative recognition of the image features in these patients is important because it precludes stapedectomy and indicates the risks in the surgery of cochlear implantation including CSF gusher and electrode insertion into IAC.

A retrospective study of laparoscopic unilateral adrenalectomy for primary hyperaldosteronism caused by unilateral adrenal hyperplasia.

OBJECTIVE: To evaluated the long-term outcomes of laparoscopic unilateral adrenalectomy for primary aldosteronism (PA) caused by unilateral adrenal hyperplasia (UAH). METHODS: One hundred and sixty-four patients who underwent laparoscopic unilateral adrenalectomy for UAH from January 2004 to December 2011 were entered in this retrospective analysis. Patients demographics, perioperative parameters, and follow-up results were recorded and analyzed statistically. RESULTS: All 164 cases suffered hypertension with biochemical evidence of hyperaldosteronism prior to operation. Hypokalemia was observed in 52/164 (37.14%) patients. UAH was proved by multi-slice computed tomography (MSCT). All operations were completed successfully without any conversions or complications. Postoperative pathology confirmed that 164 cases were cortical nodular hyperplasia, of which 4 cases coexist with medullary hyperplasia and 7 with micro-adenoma. At the median follow-up of 48 months, hypertension was cured in 88 (53.7%) patients, improved in 71 (43.3%) patients, and refractory in 5 (3.05%) patients. Hypokalemia and hyperaldosteronism were cured in all patients except re-elevation of blood pressure and plasma aldosterone in two patients 1 month after adrenalectomy. CONCLUSIONS: As an underestimated subtype of PA, UAH is accepted gradually. Laparoscopic unilateral adrenalectomy is nowadays the preferred approach to treat patients with PA caused by UAH. When adrenal venous sampling is not allowed, high-resolution MSCT is a reliable test for lateralization of aldosterone hypersecretion in carefully selected patients and 97% had either cure or improvement in blood pressure control.

[Removable appliance and closed of central incisors: report of 48 cases].

PURPOSE: To evaluate the treatment effect of orthodontic traction on embedded and inverted maxillary bending central incisors. METHODS: Fifty-two embedded and inverted maxillary central incisors without-eruption ability in 48 cases were treated by combined orthodontic traction with surgical treatment after spiral CT positioning and expanding the space, the eruption situation, gingival-breaking time and teeth pulp vitality after treatment were recorded, and the treatment effect was evaluated. RESULTS: Forty-eight cases with 52 impacted upper central incisors were treated with surgical and closed orthodontic traction, successful results were achieved in 36 cases, accounting for 69.23%; 15 cases had improvement, accounting for 28.85%; 1 case failed, accounting for 1.9%. Pulp vitality tests were all normal and there was no root absorption and ankylosis. CONCLUSIONS: Selecting appropriate cases, precise positioning with spiral CT, proper space expanding and rational application of removable traction appliance are the keys to successful treatment of impacted and inverted maxillary bending central incisors of children in mixed dentition.

[CT multiplan reconstruction images of disorder of stapes].

OBJECTIVE: To evaluate the clinical application of multi-planar reformation (MPR) for the stapes with multi-slice spinal thin-section CT in the patients with disorder of stapes before prosthetic ossicular reconstruction and their impact on surgical decision. METHODS: Axial CT scanning of temporal bone was undergone in 50 volunteers. Multiplan reformatted images included coronal, sagittal and oblique MPR, were made. All the MPR images were compared with each other in order to show which one could reveal the whole structures of stapes better. The height of stapes was measured with the oblique MPR. CT findings were evaluated in 102 cases with disorder of stapes. Prosthetic ossicular reconstruction was made in 65 cases. RESULTS: The full stapes cannot be shown in axial, coronal and sagittal MPR images, however, it was shown in oblique MPR in all the cases. The highness of stapes was (3.3 +/- 0.4) mm in the abnormal group. There was chronic otitis media in 69 cases (90 ears) and congenital abnormalities of the stapes in 33 cases (47 ears) which included the dispart of incudostapedial joint in 5 cases (7 ears), defect of head and crus of stapes in 9 cases (13 ears), defect of one side of crus in 3 cases (4 ears) and absence of the oval window in 16 cases (23 ears). CONCLUSIONS: Coronal and oblique MPR images from Axial Spiral CT Data are essential for the pre-operative planning of prosthetic ossicular reconstruction. The ship, structure or defect of stapes can be shown in the oblique MPR images. It is important for the patient to choose the type of operation.