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Testicular organoids have great potential for maintaining male fertility and even restoring male infertility. However, existing studies on generating organoids with testis-specific structure and function are scarce and come with many limitations. Research on cryopreservation of testicular organoids is even more limited, and inappropriate cryopreservation methods may result in the loss of properties in resuscitated or regenerated organoids, rendering them unsuitable for clinical or research needs. In this paper, we investigated the effects of mouse age and cell number on the self-aggregation of testicular cells into spheres in low-adsorption plates. Various media compositions, culture systems, and cell numbers were used to culture cell spheres for 14 days to form testicular organoids, and the self-organization of the organoids was assessed by histological and immunofluorescence staining. We determined the appropriate cryopreservation conditions for testicular cells, cell spheres, and tissues. Subsequently, organoids derived from cryopreserved testicular tissues, testicular cells, and testicular cell spheres were compared and evaluated by histological and immunofluorescence staining. The results indicate that testicular cell spheres consisting of 30 × 104 testicular cells from 2-week-old mice were able to form organoids highly similar to the luminal structure and cell distribution of natural mouse testicular tissues. This transformation occurred over 14 days of incubation in α-MEM medium containing 10 % knockout serum replacer (KSR) using an agarose hydrogel culture system. Additionally, the Sertoli cells were tightly connected to form a blood-testis barrier. The relative rates of tubular area, germ cells, Sertoli cells, and peritubular myoid cells were 36.985 % ± 0.695, 13.347 % ± 3.102, 47.570 % ± 0.379, and 27.406 % ± 1.832, respectively. The optimal cryopreservation protocol for primary testicular cells involved slow freezing with a cryoprotectant consisting of α-MEM with 10 % dimethyl sulfoxide (DMSO). Slow freezing with cryoprotectants containing 5 % DMSO and 5 % ethylene glycol (EG) was optimal for all different volumes of testicular cell spheres. Compared to testicular organoids generated from frozen testicular tissue and cell spheres, freezing testicular cells proved most effective in maintaining organoid differentiation characteristics and cell-cell interactions. The findings of this study contribute to a "universal" testicular organoid in vitro culture protocol with promising applications for fertility preservation and restoration in prepubertal cancer patients and adult infertile patients.
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Cryopreserved testicular tissue offers a promising method to restore fertility in male infertility patients. Current protocols rely on high concentrations of penetrating cryoprotectants (pCPAs), such as dimethyl sulfoxide (DMSO), which necessitating complex washing procedures and posing risks of toxicity. Hydrogel encapsulation presents a non-toxic alternative for cellular cryopreservation. This study investigates the effects of various types, concentrations, and thicknesses of hydrogel encapsulation on the cryopreservation of mouse testicular tissue. Testicular tissues loaded with varying concentrations of DMSO were encapsulated in alginate or gelatin-methacryloyl (GelMA) hydrogels. We evaluated hydrogels as potential CPAs to reduce pCPA concentrations and determine optimal combinations for cryopreservation. Post-cryopreservation, tissues were cultured using organ culture methods to assess spermatogenesis progression. Cryomicroscopy and differential scanning calorimetry (DSC) were used to examine ice crystal formation, melting enthalpy, and non-freezing water content in different hydrogels during cooling. Results indicate that 3â¯% alginate or 5â¯% GelMA hydrogel with thin encapsulation optimally preserves mouse testicular tissue. Using 20â¯% DMSO in 5â¯% GelMA thin encapsulation showed comparable apoptosis rates, improved morphology, higher mitochondrial activity, and enhanced antioxidant capacity compared to conventional 30â¯% DMSO without encapsulation. This suggests that hydrogel encapsulation reduces pCPA concentration by 10â¯%, thereby mitigating toxic damage. Hydrogel encapsulation can reduce basement membrane shrinkage of testicular tissue during cryopreservation. Moreover, frozen tissues remained viable with preserved germ cells after being cultured for one week on alginate methacryloyl (AlgMA) hydrogel using the gas-liquid interphase method. Cryomicroscopy and DSC studies confirmed the hydrogel's ability to inhibit ice crystal growth. In conclusion, this study introduces novel strategies for male fertility preservation and advances cryopreservation technology for clinical applications in assisted reproduction.
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Criopreservación , Crioprotectores , Hidrogeles , Testículo , Animales , Masculino , Crioprotectores/química , Crioprotectores/farmacología , Ratones , Testículo/efectos de los fármacos , Criopreservación/métodos , Hidrogeles/química , Hidrogeles/farmacología , Alginatos/química , Alginatos/farmacología , Dimetilsulfóxido/química , Dimetilsulfóxido/farmacología , Gelatina/química , Rastreo Diferencial de Calorimetría , MetacrilatosRESUMEN
Age-related changes in testicular function can impact health and well-being. The mechanisms underlying age-related testicular dysfunction, such as late-onset hypogonadism (LOH), remain incompletely understood. Using single-cell RNA sequencing on human testes with LOH, we delineated Sertoli cells (SCs) as pivotal metabolic coordinators within the testicular microenvironment. In particular, lysosomal acidity probing revealed compromised degradative capacity in aged SCs, hindering autophagy and phagocytic flux. Consequently, SCs accumulated metabolites, including cholesterol, and have increased inflammatory gene expression; thus, we termed these cells as phago-/auto-lysosomal deregulated SCs. Exposure to a high-fat diet-induced phago-/auto-lysosomal dysregulated-like SCs, recapitulating LOH features in mice. Notably, efferent ductular injection and systemic TRPML1 agonist administration restored lysosomal function, normalizing testosterone deficiency and associated abnormalities in high-fat diet-induced LOH mice. Our findings underscore the central role of SCs in testis aging, presenting a promising therapeutic avenue for LOH.
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Dieta Alta en Grasa , Hipogonadismo , Lisosomas , Células de Sertoli , Masculino , Células de Sertoli/metabolismo , Animales , Lisosomas/metabolismo , Ratones , Hipogonadismo/metabolismo , Hipogonadismo/genética , Hipogonadismo/patología , Humanos , Dieta Alta en Grasa/efectos adversos , Testículo/metabolismo , Testículo/patología , Testosterona/metabolismo , Autofagia/efectos de los fármacos , Envejecimiento/metabolismoRESUMEN
Testicular tissue culture in vitro is considered an important tool for the study of spermatogenesis and the treatment of male infertility. Although agarose hydrogel is commonly used in testicular tissue culture, the efficiency of spermatogenesis in vitro is limited. In this study, testicular tissues from adult mice were cultured using a gas-liquid interphase method based on agarose (Agarose), gelatin methacryloyl (GelMA), alginate methacryloyl (AlgMA), dextran methacryloyl (DexMA), and mixture GelMA-Agarose, AlgMA-Agarose, and DexMA-Agarose hydrogels, respectively, for 32 days in vitro. The integrity of the seminiferous tubules, the density and proportions of spermatogonia, spermatocytes, Sertoli cells, and testosterone concentrations were quantified and compared between groups. Properties of different hydrogels including compression modulus, Fourier Infrared Spectroscopy (FITR) spectra, pore size, water absorption, and water retention were tested to investigate how biochemical and physical properties of hydrogels affect the results of testicular tissue culture. The results indicate that testicular tissues cultured on AlgMA exhibited the highest seminiferous tubule integrity rate (0.835 ± 0.021), the presence of a high density of spermatocytes (2107.627 ± 232.082/mm2), and a high proportion of SOX9-positive well-preserved seminiferous tubules (0.473 ± 0.047) compared to all remaining experimental groups on day 32. This may be due to the high water content of AlgMA reducing the toxic effect of oxygen on testicular tissue. In the later period of culture, testicular tissues cultured on DexMA, not DexMA-Agarose, produced significantly more testosterone (18.093 ± 3.302 ng/mL) than the other groups, suggesting that DexMA is friendly to Leydig cells. Our study provides a new idea for the optimization of the gas-liquid interphase method for achieving in vitro spermatogenesis, facilitating the future achievement of efficient in vitro spermatogenesis in more species, including humans.
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Alginatos , Dextranos , Humanos , Masculino , Animales , Ratones , Dextranos/farmacología , Alginatos/farmacología , Gelatina/farmacología , Hidrogeles/farmacología , Sefarosa/farmacología , Espermatogénesis , Testosterona , Agua/farmacologíaRESUMEN
In recent years, the two-dimensional (2D) semiconductor α-In2Se3 has great potential for applications in the fields of electronics and optoelectronics due to its spontaneous iron electrolysis properties. Through ab initio electronic structure calculations and quantum transport simulations, the interface properties and transport properties of α-In2Se3/Au contacts with different polarization directions are studied, and a two-dimensional α-In2Se3 asymmetric metal contact design is proposed. When α-In2Se3 is polarized upward, it forms an n-type Schottky contact with Au. While when α-In2Se3 is polarized downward, it forms a p-type Schottky contact with Au. More importantly, significant rectification effect is found in the asymmetric Au/α-In2Se3/Au field-effect transistor. The carrier transports under positive and negative bias voltages are found to be dominated by thermionic excitation and tunneling, respectively. These findings provide guidance for the further design of 2D α-In2Se3-based transistors.
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Ultra-rapid cooling and rewarming rate is a critical technical approach to achieve ice-free cells during the freezing and melting process. A set of ultra-rapid solid surface freeze-thaw visualization system was developed based on a sapphire flim, and experiments on droplet freeze-thaw were carried out under different cryoprotectant components, volumes and laser energies. The results showed that the cooling rate of 1 µL mixed cryoprotectant [1.5 mol/L propylene glycol (PG) + 1.5 mol/L ethylene glycol (EG) + 0.5 mol/L trehalose (TRE)] could be 9.2×10 3 °C/min. The volume range of 1-8 µL droplets could be vitrified. After comparing the proportions of multiple cryoprotectants, the combination of equal proportion mixed permeability protectant and trehalose had the best vitrification freezing effect and more uniform crystallization characteristics. During the rewarming operation, the heating curve of glassy droplets containing gold nanoparticles was measured for the first time under the action of 400-1 200 W laser power, and the rewarming rate was up to the order of 10 6 °C/min. According to the droplet images of different power rewarming processes, the laser power range for ice-free rewarming with micron-level resolution was clarified to be 1 400-1 600 W. The work of this paper simultaneously realizes the ultra-high-speed temperature ramp-up, transient visual observation and temperature measurement of droplets, providing technical means for judging the ice free droplets during the freeze-thaw process. It is conducive to promoting the development of ultra-rapid freeze-thaw technology for biological cells and tissues.
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Nanopartículas del Metal , Vitrificación , Congelación , Criopreservación/métodos , Trehalosa , Oro , Recalentamiento , Crioprotectores , Rayos LáserRESUMEN
Chirality plays an extremely important role in nature. In this work, a highly ordered and non-clustered crystalline material UiO-88-LP was synthesized by using l-proline (l-Pro)-tuning Zr-MOF and the solvothermal method, which was then modified on the glassy carbon electrode (GCE) to construct an electrochemical chiral interface for the recognition of tryptophan (Trp) configuration. UiO-88-LP composites were characterized by scanning electron microscopy, X-ray transmission diffraction, X-ray photoelectron spectroscopy, and Fourier transform infrared (FT-IR) spectroscopy. After optimization of the experimental conditions, redox peaks for l-Trp and d-Trp were clearly observed at the UiO-88-LP/GCE electrochemical sensing interface with a peak-to-current ratio (IL/ID) of 2.47. The peak current was positively correlated with the concentration of Trp. The electrochemical recognition behavior of l-Trp and d-Trp was investigated by differential pulse voltammetry. The electrochemical characterization showed that UiO-88-LP/GCE had an enantiomeric resolution of amino acids. The recognition mechanism showed that l-Pro entering the UiO-66 molecular cage provided a site for the system to be recognized, so the purpose of recognition was achieved. The relevant data provide theoretical support for the practical application of UiO-88-LP in electrochemical sensors.
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Pleural mesothelioma (PM) with pericardial involvement is extremely rare. We now report a rare case of malignant PM with constrictive pericarditis as the first presentation. A 59-year-old male diagnosed with constrictive pericarditis underwent pericardiectomy and pericardial pathology revealed mesothelial hyperplasia. Eight months after surgery, the patient was admitted to the hospital with chest tightness and wheezing for 5 days. Computed tomography scan of the chest showed a left lung expansion insufficiency, limited bilateral pleural thickening, pericardial thickening with a small amount of pericardial effusion, and multiple enlarged lymph nodes in the mediastinum, bilateral supraclavicular fossa, bilateral cervical roots, and right axilla. The pleural malignancy should be possibly considered. Pathology after pleural puncture showed malignant PM. Pathology after left supraclavicular lymph node puncture biopsy showed metastatic malignant mesothelioma. The diagnosis of this patient was clear. Although malignant PM rarely involves the pericardial constriction, we cannot ignore the fact that malignant PM involves the pericardium. The patient has been diagnosed with constrictive pericarditis, accompanied by pleural thickening and pleural effusion. Without other pathogenic factors, pleural biopsy should be aggressively performed in patients with constrictive pericarditis to determine the cause.
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Genetic causation for the majority of non-obstructive azoospermia (NOA) remains unclear. Mutations in synaptonemal complex (SC)-associated genes could cause meiotic arrest and NOA. Previous studies showed that heterozygous truncating variants in SYCP2 encoding a protein essential for SC formation, are associated with non-obstructive azoospermia and severe oligozoospermia. Herein, we showed a homozygous loss-of-function variant in SYCP2 (c.2689_2690insT) in an NOA-affected patient. And this variant was inherited from heterozygous parental carriers by natural reproduction. HE, IF, and meiotic chromosomal spread analyses demonstrated that spermatogenesis was arrested at the zygotene stage in the proband with NOA. Thus, this study revealed that SYCP2 associated with NOA segregates in an autosomal recessive inheritance pattern, rather than an autosomal dominant pattern. Furthermore, our study expanded the knowledge of variants in SYCP2 and provided new insight into understanding the genetic etiology of NOA.
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Azoospermia , Masculino , Humanos , Azoospermia/genética , Mutación del Sistema de Lectura , Mutación , Espermatogénesis/genética , Proteínas de Unión al ADN/genética , Proteínas de Ciclo Celular/genéticaRESUMEN
AlGaN is an important material for deep ultraviolet optoelectronic devices and electronic devices. The phase separation on the AlGaN surface means small-scale compositional fluctuations of Al, which is prone to degrade the performance of devices. In order to study the mechanism of the surface phase separation, the Al0.3Ga0.7N wafer was investigated by the scanning diffusion microscopy method based on the photo-assisted Kelvin force probe microscope. The response of the surface photovoltage near the bandgap was quite different for the edge and the center of the island on the AlGaN surface. We utilize the theoretical model of scanning diffusion microscopy to fit the local absorption coefficients from the measured surface photovoltage spectrum. During the fitting process, we introduce as and ab parameters (bandgap shift and broadening) to describe the local variation of absorption coefficients α(as, ab, λ). The local bandgap and Al composition can be calculated quantitatively from the absorption coefficients. The results show that there is lower bandgap (about 305 nm) and lower Al composition (about 0.31) at the edge of the island, compared with those at the center of the island (about 300 nm for bandgap and 0.34 for Al composition). Similar to the edge of the island, there is a lower bandgap at the V-pit defect which is about 306 nm corresponding to the Al composition of about 0.30. These results mean Ga enrichment both at the edge of the island and the V-pit defect position. It proves that scanning diffusion microscopy is an effective method to review the micro-mechanism of AlGaN phase separation.
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The development of low-dimensional multifunctional devices has become increasingly important as the size of field-effect transistors decreases. In recent years, the two-dimensional (2D) semiconductor In2Se3 has emerged as a promising candidate for applications in the fields of electronics and optoelectronics owing to its remarkable spontaneous polarization properties. Through first-principles calculations, the effects of the polarization direction and biaxial tensile strain on the electronic and contact properties of In2Se3/Au heterostructures are investigated. The contact type of In2Se3/Au heterostructures depends on the polarization direction of In2Se3. The more charge transfers from the metal to the space charge region, the biaxial tensile strain increases. Moreover, the upward polarized In2Se3 in contact with Au maintains a constant n-type Schottky contact as the biaxial tensile strain increases, with a barrier height Φ SB,n of only 0.086 eV at 6% strain, which is close to ohmic contact. On the other hand, the downward polarized In2Se3 in contact with Au can be transformed from p-type to n-type by applying a biaxial tensile strain. Our calculation results can provide a reference for the design and fabrication of In2Se3-based field effect transistors.
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Chirality plays a key role in many fields of natural sciences as well as life sciences. Chiral materials are widely developed and used for electrochemical chiral recognition. In recent years, carbon quantum dots (CQDs) have been widely used as a novel carbon nanomaterial due to their excellent charge transfer properties, good biocompatibility, and low cost. The special structure of π-conjugated porphyrin attracts attention. Supramolecular self-assembly shows a way to construct chiral materials by self-assembling simple molecules into chiral composites. Herein, we demonstrate the self-assembly of achiral porphyrins induced by chiral carbon quantum dots assembled from L- and or D-tryptophan (L- and or D-Trp) with carbon quantum dots, resulting in 5,10,15,20-tetrakis (4-carboxyPheyl) (TCPP) self-assembled structure. The electrochemical chiral recognition of chiral self-assembled materials was studied using Phenylalanine (Phe) enantiomer as a chiral analyte. Electrochemical chiral recognition results showed that the chiral self-assembled materials induced by chiral templates have a good ability to discriminate Phe enantiomers. Therefore, this research provides a new idea for the synthesis of chiral composites and further expands applications to electrochemical chiral recognition.
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Nanoestructuras , Porfirinas , Porfirinas/química , Fenilalanina/química , Triptófano , CarbonoRESUMEN
Rhinacanthin C (RC) is a naphthoquinone ester with an anti-inflammatory activity extracted from Rhinacanthus nasutus (L.) Kurz (Rn). It has been proven to improve hyperglycemia and hyperlipidemia, but the prevention and mechanism of RC in nonalcoholic fatty liver disease (NAFLD) are not clear. In the current study, we first extracted RC from Rn using ethyl acetate and identified it by HPLC, MS, and NMR. At the same time, molecular docking analysis of RC with AMPK and SREBP-1c was performed using AutoDock software. In addition, the mouse model of NAFLD was induced by a high-fat diet in vivo, and low, medium, and high concentrations of RC were used for intervention. The results showed that RC significantly reduced the body mass and liver body coefficient of NAFLD mice, inhibited liver inflammation and fat accumulation, and improved insulin resistance. Further studies showed that RC significantly reduced the levels of serum leptin and resistin, upregulated the expression levels of adiponectin and adiponectin receptor in the liver, and inhibited the expression levels of MCP-1, TNF-α, and IL-6. In terms of mechanism, RC upregulates the expression of p-AMPK and SIRT1 and downregulates the expression of p-p65, SREBP-1c, Fas, Acc-α, PPAR-γ, and SCD1. These studies suggest that RC improves insulin resistance and lipid accumulation in NAFLD by activating the AMPK/SIRT1 and SREBP-1c/Fas/ACC pathways, respectively.
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Malignant peripheral nerve sheath tumor (MPNST) is a rare tumor that can develop on the lining of nerves and within the network of nerve fibers in different organs, and it is commonly found in the head and neck, limbs, and trunk. These tumors can occur in patients of any age. They most commonly occur in adults aged 20 to 50 years; however, fewer cases of this tumor in children have been reported. To date, no neonatal case of MPNST in the nasal cavity has been reported. Here, we report the case of a 4-day-old female newborn who presented with a nasal mass that re-enlarged after surgery and was diagnosed as MPNST of the nasal cavity on the basis of pathological results. This is the first report of MPNST in the nasal cavity of a neonate. Differential diagnosis and treatment of nasal masses have been proposed in the related literature.
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Neoplasias de la Vaina del Nervio , Neurofibrosarcoma , Adulto , Niño , Recién Nacido , Humanos , Femenino , Neoplasias de la Vaina del Nervio/diagnóstico , Neoplasias de la Vaina del Nervio/cirugía , Neoplasias de la Vaina del Nervio/patología , Cavidad Nasal/patologíaRESUMEN
OBJECTIVES: To establish a system for regulating the gene expression of embryonic mouse cerebral cortex neural stem cells (NSCs) using in utero electroporation (IUE). METHODS: At embryonic day 14.5, the mouse cerebral cortex NSCs were electro-transfected with the pCIG plasmid injected into the ventricle of the mouse embryo. At embryonic day 16.5 or day 17.5, embryonic mouse brain tissues were collected to prepare frozen sections. Immunofluorescence staining was used to observe the proliferation, apoptosis, division, directional differentiation, migration, and maturation of NSCs. RESULTS: The differentiation of NSCs into intermediate progenitors, the proliferation and apoptosis of NSCs, and the morphological development of radial axis of radial glial cells were observed at embryonic day 16.5. The differentiation of NSCs into neurons in layers V-VI of the cerebral cortex, the migration of NSCs to the lateral cerebral cortex, the development of dendrites of migrating neurons, and the maturation of neurons were observed at embryonic day 17.5. CONCLUSIONS: The system for regulating the gene expression of embryonic mouse cerebral cortex NSCs can be established using IUE, which is useful for the study of neural development related to the proliferation, apoptosis, division, directional differentiation, migration and maturation of NSCs in the cerebral cortex.
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Células-Madre Neurales , Animales , Corteza Cerebral/metabolismo , Electroporación , Expresión Génica , Ratones , Neuronas/metabolismoRESUMEN
The corpus cavernosum is the most important structure for penile erection, and its dysfunction causes many physiological and psychological problems. However, its cellular heterogeneity and signalling networks at the molecular level are poorly understood because of limited access to samples. Here, we profile 64,993 human cavernosal single-cell transcriptomes from three males with normal erection and five organic erectile dysfunction patients. Cell communication analysis reveals that cavernosal fibroblasts are central to the paracrine signalling network and regulate microenvironmental homeostasis. Combining with immunohistochemical staining, we reveal the cellular heterogeneity and describe a detailed spatial distribution map for each fibroblast, smooth muscle and endothelial subcluster in the corpus cavernosum. Furthermore, comparative analysis and related functional experiments identify candidate regulatory signalling pathways in the pathological process. Our study provides an insight into the human corpus cavernosum microenvironment and a reference for potential erectile dysfunction therapies.
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Disfunción Eréctil , Disfunción Eréctil/genética , Humanos , Masculino , Músculo Liso/patología , Erección Peniana/fisiología , Pene , Transcriptoma/genéticaRESUMEN
With the increase in extremely low birth weight (ELBW) infants, their outcome attracted worldwide attention. However, in China, the related studies are rare. The hospitalized records of ELBW infants discharged from twenty-six neonatal intensive care units in Guangdong Province of China during 2008-2017 were analyzed. A total of 2575 ELBW infants were enrolled and the overall survival rate was 55.11%. From 2008 to 2017, the number of ELBW infants increased rapidly from 91 to 466, and the survival rate improved steadily from 41.76% to 62.02%. Increased survival is closely related to birth weight (BW), regional economic development, and specialized hospital. The incidence of complications was neonatal respiratory distress syndrome (85.2%), oxygen dependency at 28 days (63.7%), retinopathy of prematurity (39.3%), intraventricular hemorrhage (29.4%), necrotizing enterocolitis (12.0%), and periventricular leukomalacia (8.0%). Among the 1156 nonsurvivors, 90.0% of infants died during the neonatal period (≤ 28 days). A total of 768 ELBW infants died after treatment withdrawal, for reasons of economic and/or poor outcome. The number of ELBW infants is increasing in Guangdong Province of China, and the overall survival rate is improving steadily.
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Enterocolitis Necrotizante , Enfermedades del Prematuro , Estudios de Cohortes , Enterocolitis Necrotizante/epidemiología , Humanos , Lactante , Mortalidad Infantil , Recien Nacido con Peso al Nacer Extremadamente Bajo , Recién Nacido , Enfermedades del Prematuro/epidemiologíaRESUMEN
BACKGROUND: Non-obstructive azoospermia (NOA) is the most severe disease in male infertility, but the genetic causes for majority of NOA remain unknown. METHODS: Two Chinese NOA-affected patients were recruited to identify the genetic causal factor of infertility. Whole-exome sequencing (WES) was conducted in the two patients with NOA. Sanger sequencing and CNV array were used to ascertain the WES results. Hematoxylin and eosin (H&E) staining and immunofluorescence (IF) were carried out to evaluate the stage of spermatogenesis arrested in the affected cases. RESULTS: Novel heterozygous deletion (LOH) within SYCE1 (seq[GRCh37] del(10)(10q26.3)chr10:g.135111754_135427143del) and heterozygous loss of function (LoF) variant in SYCE1 (NM_001143763: c.689_690 del:p.F230fs) were identified in one NOA-affected patient. While homozygous deletion within SYCE1 (seq[GRCh37] del(10)(10q26.3)chr10:g.135340247_135379115del) was detected in the other patient with meiotic arrest. H&E and IF staining demonstrated that the spermatogenesis was arrested at pachytene stage in the two patients with NOA, suggesting these two novel CNVs within SYCE1 could lead to meiotic defect and NOA. CONCLUSIONS: We identified that two novel CNVs within SYCE1 are associated with meiotic arrest and male infertility. Thus, our study expands the knowledge of variants in SYCE1 and provides a new insight to understand the genetic etiologies of NOA.
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Azoospermia , Proteínas de Unión al ADN , Infertilidad Masculina , Azoospermia/genética , Variaciones en el Número de Copia de ADN , Proteínas de Unión al ADN/genética , Homocigoto , Humanos , Infertilidad Masculina/genética , Masculino , Mutación , Eliminación de SecuenciaRESUMEN
Renal injury in lupus nephritis (LN) does not manifest as one uniform entity. The clinical presentation, management, and prognosis of membranous LN (MLN) differ from that of the proliferative LN (PLN). Differentiating the molecular mechanisms involved in MLN and PLN and discovering the reliable biomarkers for early diagnosis and target therapy are important. We compared the kidney protein expression patterns of 11 pure MLN and 12 pure PLN patients on formalin-fixed paraffin-embedded (FFPE) kidney tissues using label-free liquid chromatography-mass spectrometry (LC-MS) for quantitative proteomics analysis. FunRich software was used to identify proteins in differentially expressed pathways. Quantitative comparisons of differentially expressed proteins in each patient were further analyzed based on protein intensity levels determined by LC-MS. The protein-protein interaction (PPI) network of the differentially expressed genes (DEGs) was established through Search Tool for the Retrieval of Interacting Genes database (STRING) website, visualized by Cytoscape. A total of 5112 proteins were identified. In total, 12 significantly upregulated (fold change ≥2, p < 0.05) proteins were identified in the MLN group and 220 proteins (fold change ≥2, p < 0.05) were upregulated in the PLN group. Further analysis showed that the most significant upregulated pathway involved in MLN was histone deacetylase (HDAC) class I pathway, and the three most significant upregulated pathways in PLN were interferon signaling, interferon gamma signaling, and the immune system. Next, we selected sirtuin-2 (SIRT2) in MLN, and vascular cell adhesion protein 1 (VCAM1) and Bcl-xl in PLN for further mass spectrometry (MS) intensity and PPI analysis. SIRT2 expression was significantly increased in the MLN group compared with the PLN group, and VCAM1, Bcl-xl expression was significantly increased in the PLN group compared with the MLN group, based on MS intensity. These results may help to improve our understanding of the underlying molecular mechanisms of MLN and PLN and provide potential targets for the diagnosis and treatment of different subclasses of LN.