Precise Correction of Lhcgr Mutation in Stem Leydig Cells by Prime Editing Rescues Hereditary Primary Hypogonadism in Mice.

Hereditary primary hypogonadism (HPH), caused by gene mutation related to testosterone synthesis in Leydig cells, usually impairs male sexual development and spermatogenesis. Genetically corrected stem Leydig cells (SLCs) transplantation may provide a new approach for treating HPH. Here, a novel nonsense-point-mutation mouse model (LhcgrW495X ) is first generated based on a gene mutation relative to HPH patients. To verify the efficacy and feasibility of SLCs transplantation in treating HPH, wild-type SLCs are transplanted into LhcgrW495X mice, in which SLCs obviously rescue HPH phenotypes. Through comparing several editing strategies, optimized PE2 protein (PEmax) system is identified as an efficient and precise approach to correct the pathogenic point mutation in Lhcgr. Furthermore, delivering intein-split PEmax system via lentivirus successfully corrects the mutation in SLCs from LhcgrW495X mice ex vivo. Gene-corrected SLCs from LhcgrW495X mice exert ability to differentiate into functional Leydig cells in vitro. Notably, the transplantation of gene-corrected SLCs effectively regenerates Leydig cells, recovers testosterone production, restarts sexual development, rescues spermatogenesis, and produces fertile offspring in LhcgrW495X mice. Altogether, these results suggest that PE-based gene editing in SLCs ex vivo is a promising strategy for HPH therapy and is potentially leveraged to address more hereditary diseases in reproductive system.

AAV-mediated gene therapy produces fertile offspring in the Lhcgr-deficient mouse model of Leydig cell failure.

Leydig cell failure (LCF) caused by gene mutation results in testosterone deficiency and infertility. Serum testosterone levels can be recovered via testosterone replacement; however, established therapies have shown limited success in restoring fertility. Here, we use a luteinizing hormone/choriogonadotrophin receptor (Lhcgr)-deficient mouse model of LCF to investigate the feasibility of gene therapy for restoring testosterone production and fertility. We screen several adeno-associated virus (AAV) serotypes and identify AAV8 as an efficient vector to drive exogenous Lhcgr expression in progenitor Leydig cells through interstitial injection. We observe considerable testosterone recovery and Leydig cell maturation after AAV8-Lhcgr treatment in pubertal Lhcgr-/- mice. Of note, this gene therapy partially recovers sexual development, substantially restores spermatogenesis, and effectively produces fertile offspring. Furthermore, these favorable effects can be reproduced in adult Lhcgr-/- mice. Our proof-of-concept experiments in the mouse model demonstrate that AAV-mediated gene therapy may represent a promising therapeutic approach for patients with LCF.

Serum miR-21 predicts the prognosis of patients with primary gastrointestinal diffuse large B-cell lymphoma.

BACKGROUND: Primary gastrointestinal diffuse large B-cell lymphoma (PGI-DLBCL) lacks specific clinical manifestations and its malignancy renders prognostication and choice of treatment strategy difficult. The aim of this study was to evaluate microRNA (miR)-21 as potential non-invasive biomarkers for prognosis in PGI-DLBCL patients. METHODS: Serum miR-21 expression in de novo PGI-DLBCL patients, consecutively enrolled for this study, was detected by quantitative real-time polymerase chain reaction (qRT-PCR). Relative expression was calculated using the comparative Ct method. Statistical significance was determined using the Mann-Whitney rank sum and Fisher's exact test. Survival analysis was conducted using the Kaplan-Meier method. RESULTS: Compared with healthy controls, serum miR-21 levels were significantly elevated in the PGI-DLBCL patients (n=156). The high expression level of serum miR-21 at diagnosis was associated with worse progression-free survival (PFS) (30 (9-42) vs 42 (12-52) months in high and low miR-21 groups) and overall survival (OS) (35 (15-52) vs 48 (17-61) months in high and low miR-21 groups) and was an independent risk factor for PFS and OS (hazard ratios 4.345 and 3.311, respectively). Furthermore, Bcl-2, Bcl-6 and Ki-67 were independently and positively associated with miR-21 expression. CONCLUSIONS: Our results suggest that miR-21 is a potential prognostic marker to predict clinical outcomes in PGI-DLBCL patients and a high miR-21 level is associated with poor outcomes.

Early hepatoid adenocarcinoma of the stomach with signet ring cell carcinoma: A case report and clinicopathological features.

Background: Hepatoid adenocarcinoma of the stomach (HAS) is a rare subtype of gastric cancer with poor prognosis, and its clinicopathological features are not well understood, so the pathology from the clinical biopsy is easily misdiagnosed, especially for special or atypical HAS. We present an extremely rare early HAS with signet ring cell carcinoma and evaluate its clinicopathological features. Case presentation: A 51-year-old female patient of Chinese Han ethnicity with upper abdominal pain for 5 years and worsened abdominal pain for 1 month was admitted to our hospital. Esophagogastroduodenoscopy showed a submucosal tumor-like elevated lesion with central depression in the greater curvature of the junction between the antrum and body. Histopathological examination from the biopsy revealed medium-low-differentiation adenocarcinoma with signet ring cell carcinoma. Radical gastrectomy was performed, and the final diagnosis was early HAS with signet ring cell carcinoma. Conclusions: HAS with signet ring cell carcinoma is a special type of HAS and extremely rare. It is first presented for this extremely rare type of HAS, which contributes to strengthen the understanding for the clinicopathological characteristics of HAS and especially promote early detection of HAS.

Medullary thyroid carcinoma combined with papillary thyroid carcinoma: case report and literature review.

We investigated the clinicopathologic features, immunophenotype, (differential) diagnosis, pathogenesis, treatment, and follow-up of medullary thyroid carcinoma (MTC) combined with papillary thyroid carcinoma (PTC). A retrospective analysis of the clinical and pathologic features and immunophenotype was conducted in a patient with MTC and PTC. Relevant literature was also reviewed. Results of thyroid fine needle aspiration indicated malignant tumor in the right lobe of the thyroid, suggesting PTC; further analysis by biopsy confirmed this diagnosis. The left lobe exhibited MTC. Tumor metastases were absent from the lymph nodes of the left central area (0/2), and no tumor was present in the thymic tissue. In the right lobe and isthmus, PTC was observed, with a maximum infiltration diameter of 0.8 cm, and tumor metastases were absent from lymph nodes of the right central area (0/3). Immunohistochemistry of the left lobe was positive for calcitonin, CK, TTF-1, CD56, CgA, and Congo red, but negative for CK19, thyroglobulin, galectin-3, MC, and CEA, with a Ki-67 proliferation index of 1%. The right lobe was positive for CK19, galectin-3, and MC, but negative for CD56. The V600E mutation was detected in BRAF. MTC combined with PTC is a rare thyroid tumor. This condition is diagnosed mainly based on morphology, immunophenotyping, and molecular detection. It must be distinguished from other malignancies, such as thyroid follicular tumors, undifferentiated carcinoma, poorly differentiated carcinoma, transparent stellate tumor, and mixed PTC/MTC. Surgery and post-operative drug administration currently constitute the preferred treatments.

Hollow Mesoporous Carbon Sphere Loaded Ni-N4 Single-Atom: Support Structure Study for CO2 Electrocatalytic Reduction Catalyst.

Single-atom catalysts have become a hot spot because of the high atom utilization efficiency and excellent activity. However, the effect of the support structure in the single-atom catalyst is often unnoticed in the catalytic process. Herein, a series of carbon spheres supported Ni-N4 single-atom catalysts with different support structures are successfully synthesized by the fine adjustment of synthetic conditions. The hollow mesoporous carbon spheres supported Ni-N4 catalyst (Ni/HMCS-3-800) exhibits superior catalytic activity toward the electrocatalytic CO2 reduction reaction (CO2 RR). The Faradaic efficiency toward CO is high to 95% at the potential range from -0.7 to -1.1 V versus reversible hydrogen electrode and the turnover frequency value is high up to 15 608 h-1 . More importantly, the effect of the geometrical structures of carbon support on the CO2 RR performance is studied intensively. The shell thickness and compactness of carbon spheres regulate the chemical environment of the doped-N species in the carbon skeleton effectively and promote CO2 molecule activation. Additionally, the optimized mesopore size is beneficial to improve diffusion and overflow of the substance, which enhances the CO2 adsorption capacity greatly. This work provides a new consideration for promoting the catalytic performance of single-atom catalysts.

ALK-1-positive inflammatory myofibroblastic tumor of the thyroid complicated by Hashimoto's thyroiditis: report of a rare case and a literature review.

BACKGROUND: Inflammatory myofibroblastic tumors (IMTs) of the thyroid are extremely rare soft-tissue tumors. In the literature, IMTs are sometimes called plasma cell granulomas (PCGs) or inflammatory pseudotumors, which often causes ambiguity. To date, 17 cases of PCGs and five cases of thyroid IMTs have been reported. These cases reveal that IMTs of the thyroid are often negative for the anaplastic lymphoma kinase (ALK-1) gene. To provide further information on this rare lesion, we present a case of an ALK-1-positive thyroid IMT and a review of IMTs of the thyroid. CASE PRESENTATION: A 34-year-old Chinese woman presented with a painless neck mass that had persisted for over a month. Ultrasonography revealed a 4.28 × 2.53 cm2 hypoechoic mass, in the left lobe of the thyroid gland. Serum levels of thyroglobulin and anti-thyroglobulin antibodies were high. Subsequently, left lobectomy was performed. Macroscopically, the lesion was a gray-brown nodular mass with a partial envelope. Histologically, two different lesion types were observed. The first lesion showed classic spindle cell proliferation, with spindle cells arranged in fascicles, accompanied by mature inflammatory cells. The other lesion showed a large number of infiltrating lymphocytes, with lymphoid follicles in the remaining thyroid gland, which was atrophic. Immunohistochemical staining showed that the spindle cells were negative for CK19, CyclinD1, Gelectin-3, EMA, CD34, S100, Bcl-2, and STAT-6, but strongly positive for ALK-1, vimentin, and TTF1. CK was focally expressed, and the Ki-67 index was 5%. A diagnosis of IMT was proposed according to immunohistochemical findings and morphology. Hashimoto's thyroiditis was confirmed according to serum levels of thyroglobulin and anti-thyroglobulin antibodies and morphology. The patient did not receive adjuvant therapy. She remained alive without disease recurrence for 10 months after lobectomy. CONCLUSIONS: IMTs should be considered in the diagnosis when spindle cell proliferation accompanied by mature inflammatory cells is observed, spindle cells are mildly atypical, and myofibroblast differentiation is present in the thyroid. A uniform diagnostic term is crucial to avoid ambiguity. Clinicians and pathologists should be aware of the necessity for long-term follow-up, especially in ALK-positive cases. The therapeutic potential of ALK-1 positivity should be explored further.

Ultra-small Pd nanoparticles derived from a supramolecular assembly for enhanced electrochemical reduction of CO2 to CO.

Ultra-small Pd nanoparticles were successfully prepared through reduction of a supramolecular assembly formed between the macrocyclic decamethylcucurbit[5]uril (Me10CB[5]) and [PdCl4]2- anions via H-bonds. The final Pd NPs exhibit excellent activity and stability toward the electrochemical reduction of CO2 to CO. This work provides a new strategy for the preparation of nanocatalysts.

Giant recurrent mixed-type liposarcoma of the retroperitoneum: report of a case and review of literature.

Retroperitoneal liposarcoma is a rare tumor with an incidence of 2.5 per million individuals, especially tumors of mixed histologic pattern. We present a case of a 63-year-old woman with a history of slowly increasing abdominal volume over 2 years. The diagnosis of giant and multiple retroperitoneal mass suspected of liposarcoma was confirmed by computed tomography. The patient underwent resection of 7 tumor masses together weighing 5 kg. The microscopic diagnosis was mixed-type liposarcoma of the retroperitoneum. 8 months after surgery, the patient suffered multiple metastases in the liver and abdominal wall, and upper digestive tract hemorrhage. Although this type of tumor is rarely seen, tumor tissue should be thoroughly gathered and analyzed on histologic examination to reach final diagnosis. Knowledge of the subtype of liposarcoma is important for proper prognosis and treatment of the patient. To the best of our knowledge, this is the first report of mixed-type retroperitoneal liposarcoma with three components described in the English literature.

Carvacrol attenuates traumatic neuronal injury through store-operated Ca(2+) entry-independent regulation of intracellular Ca(2+) homeostasis.

Searching for effective pharmacological agents for traumatic brain injury (TBI) treatment has largely been unsuccessful. The transient receptor potential melastatin 7 (TRPM7), a TRP channel that is essential for embryonic development, has been shown to mediate ischemic neuronal injury in vivo and in vitro, but global deletion of TRPM7 in mice is lethal. Here, carvacrol was used to investigate the protective effect of TRPM7 inhibition in an in vitro traumatic neuronal injury model. Carvacrol (0.5 and 1 mM) reduced lactate dehydrogenase (LDH) release, apoptosis and caspase-3 activation after traumatic injury in cortical neurons. These neuroprotective effects were accompanied by alleviated cytoplasmic calcium levels as measured by calcium imaging. In contrast, the thapsigargin (TG) induced store-operated calcium entry (SOCE) and the expression of SOCE related proteins in neurons were not altered by carvacrol treatment. The involvement of TRPM7 sensitive calcium influx in our in vitro model was confirmed by the results that bradykinin induced calcium influx was prevented by carvacrol in neurons. Furthermore, carvacrol significantly inhibited the induction of neuronal nitric oxide synthase (nNOS) after traumatic injury, and treatment with carvacrol and the nNOS inhibitor NLPA together had no extra effect on calcium concentration and neuronal injury. Thus, inhibition of TRPM7 function by carvacrol protects against traumatic neuronal injury, and might be a potential drug development strategy for the treatment of TBI.