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1.
Biopreserv Biobank ; 2020 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-32423228

RESUMO

Recently, many studies have found that late embryogenesis abundant (LEA) proteins could protect cells from drought, high salinity, and other stress conditions. Because LEA proteins maintain the integrity and stability of cell membranes, LEA proteins increase the cell's tolerance to dehydration stress, and reduce the osmotic and freezing damage during freezing. Whether LEA proteins could reduce cryopreservation damage and improve cell viability remains to be confirmed. In this study, we purified the recombinant AavLEA1 proteins, examined their thermal solubility and the effect of AavLEA1 proteins on the osmotic stress of cells, and studied the effects of the AavLEA1 protein on cryopreservation of human umbilical cord matrix mesenchymal stem cells (hUCM-MSCs). We utilized three concentrations of AavLEA1 protein (0.1, 0.5, and 2 mg/mL) to cryopreserve hUCM-MSCs and analyzed cell viability and apoptosis of MSCs after slow-cooling cryopreservation. We also examined the cryopreservation effect of AavLEA1 protein on hUCM-MSCs survival with 0%, 2%, 5%, and 10% (v/v) dimethyl sulfoxide (DMSO). We found that the survival rate of hUCM-MSCs supplemented with AavLEA1 protein was significantly higher than that of MSCs cryopreserved with low concentration of DMSO solution, and the apoptosis and necrosis rates were correspondingly reduced. In conclusion, recombinant AavLEA1 protein can improve the efficiency of MSC cryopreservation, increase the hUCM-MSCs viability, and partly replace DMSO during cryopreservation.

2.
Cell Transplant ; 29: 963689720918300, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32425116

RESUMO

Given the crucial role of microRNAs in the cellular proliferation of various types of cancers, we aimed to analyze the expression and function of a cellular proliferation-associated miR-188-5p in papillary thyroid carcinoma (PTC). Here we demonstrate that miR-188-5p is downregulated in PTC tumor tissues compared with the associated noncancerous tissues. We also validate that the miR-188-5p overexpression suppressed the PTC cancer cell proliferation. In addition, fibroblast growth factor 5 (FGF5) is observed to be downregulated in the PTC tumor tissues compared with the associated noncancerous tissues. Subsequently, FGF5 is identified as the direct functional target of miR-188-5p. Moreover, the silencing of FGF5 was found to inhibit PTC cell proliferation, which is the same pattern as miR-188-5p overexpression. These results suggest that miR-188-5p-associated silencing of FGF5 inhibits tumor cell proliferation in PTC. It also highlights the importance of further evaluating miR-188-5p as a potential biomarker and therapy target in PTC.

4.
Surg Endosc ; 2020 May 19.
Artigo em Inglês | MEDLINE | ID: mdl-32430528

RESUMO

BACKGROUND: The surgery for esophageal cancer arising after prior gastrectomy is technically difficult with high morbidity and mortality. Endoscopic submucosal dissection (ESD) is a minimally invasive endoscopic treatment for superficial SCC with high curative resection rate. But few studies are concerned about ESD under these circumstances. The aim of this study was to elucidate the short- and long-term outcomes of ESD for superficial esophageal squamous cell cancer (SCC) in patients with prior gastrectomy. METHODS: From January 2009 to January 2019, 37 patients with prior gastrectomy who underwent ESD for superficial esophageal SCC were retrospectively enrolled at the Zhongshan Hospital, Fudan University in Shanghai, China. Rates of en bloc resection, complete resection, curative resection, incidence of postoperative bleeding, perforation and postoperative stricture were evaluated as short-term outcomes. Overall survival, and local recurrence-free survival were evaluated as long-term outcomes. RESULTS: The rate of en bloc resection, complete resection and curative resection were 94.6%, 86.5% and 78.4%, respectively. No perforation was observed. 1 (2.7%) patient experienced postoperative bleeding. During the median observation of 43 months, 3 (8.6%) patients experienced esophageal stricture, successfully managed by balloon dilation. 3(8.6%) patients had local recurrence after ESD with 5-year local recurrence-free survival rate of 91.4%. During the observation period, 4 patients died of other reasons. The 1, 3, 5-year overall survival rates were 97.1%, 97.1% and 91.4%. CONCLUSIONS: The short-term outcomes indicate ESD is technically difficult with lower resection completeness in patients after gastrectomy, while the long-term outcomes are rather favorable.

5.
Cell Death Dis ; 11(5): 400, 2020 May 26.
Artigo em Inglês | MEDLINE | ID: mdl-32457294

RESUMO

DNA-dependent protein kinase catalytic subunit (DNA-PKcs) is the core component of DNA-PK complex in the non-homologous end-joining (NHEJ) repair of DNA double-strand breaks, and its activity is strictly controlled by DNA-PKcs phosphorylation. The ubiquitin-like protein, NEDD8 is involved in regulation of DNA damage response, but it remains mysterious whether and how NEDD8-related neddylation affects DNA-PKcs and the NHEJ process. Here, we show that DNA-PKcs is poly-neddylated at its kinase domain. The neddylation E2-conjugating enzyme UBE2M and E3 ligase HUWE1 (HECT, UBA, and WWE domain containing E3 ubiquitin protein ligase 1) are responsible for the DNA-PKcs neddylation. Moreover, inhibition of HUWE1-dependent DNA-PKcs neddylation impairs DNA-PKcs autophosphorylation at Ser2056. Finally, depletion of HUWE1-dependent DNA-PKcs neddylation reduces the efficiency of NHEJ. These studies provide insights how neddylation modulates the activity of NHEJ core complex.

6.
Microb Pathog ; 145: 104225, 2020 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-32353581

RESUMO

Regulatory T cells (Tregs) play an essential role during homeostasis and tolerance of the immune system. Based on our previous study that exposure of pregnant rats to staphylococcal enterotoxin B (SEB) can alter the percentage of CD4/CD8 subsets in the thymus of the offspring, in this study, we focus on the influence of exposure of pregnant rats to SEB on number, function and response of Tregs in the thymus of the offspring. Pregnant rats at gestational day of 16 were intravenously injected with 15 µg SEB and the thymuses of the neonatal and adult offspring were harvested for this study. We found that exposure of pregnant rats to SEB could significantly increase the absolute number of Tregs and the FoxP3 expression level in the thymus of not only neonatal but also adult offspring. Re-exposure of adult offspring to SEB remarkably reduced the suppressive capacity of Tregs to CD4+ T cells and the expression levels of TGF-ß and IL-10 in the thymus, but had no effect on production of IL-4 and IFN-γ. Furthermore, it also notedly decreased the absolute number of Tregs and the FoxP3 expression level. These data suggest that prenatal exposure of pregnant rats to SEB attenuates the response of increased Tregs to re-exposure to SEB in the thymus of adult offspring.

7.
J Cell Physiol ; 2020 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-32391604

RESUMO

Facial paralysis can result in severe implications for patients. A good prognosis depends on the degree of nerve regeneration. Schwann cells (SCs) play an important role in facial nerve development and regeneration through migration. Forkhead box C1 (Foxc1), a member of the forkhead transcription factor family, is implicated in cell migration. However, the role of Foxc1 in the progression after facial nerve crush remains unknown. Our aim was to evaluate the effect of Foxc1 overexpression on SC migration and recovery of facial nerves after crush injury. The rat facial nerve crush injury model was established through the use of unilateral surgery. The results showed that the expression of Foxc1 was increased in the surgery group compared to that of the control group. SCs were isolated from the sciatic nerves and cultured. Foxc1, delivered by an adeno-associated virus in vivo, or adenovirus in vitro, both induced overexpression of Foxc1, and increased the expression of CXCL12 and ß-catenin. After the transfection of Foxc1, the migration of SC was increased both in vitro and in vivo, was reduced by the inhibition of CXCL12 or ß-catenin. The facial nerve function and the nerve axon remyelination of the rats transfected with Foxc1 were significantly improved after nerve crush injury. Overall, the results demonstrated that overexpression of Foxc1 promoted SC migration by regulating CXCL12 via the Wnt/ß-catenin pathway, thus contributing to improved facial nerve function after crush injury.

8.
Neural Plast ; 2020: 5701042, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32377180

RESUMO

Chronic shoulder pain (CSP) is the third most common musculoskeletal problem. For maximum treatment effectiveness, most acupuncturists usually choose acupoint in the nonpainful side, to alleviate pain or improve shoulder function. This method is named opposite needling, which means acupuncture points on the right side are selected for diseases on the left side and vice versa. However, the underlying neural mechanisms related to treatment are currently unclear. The purpose of this study was to determine whether different mechanisms were observed with contralateral and ipsilateral acupuncture at Tiaokou (ST 38) in patients with unilateral CSP. Twenty-four patients were randomized to the contralateral acupuncture group (contra-group) and the ipsilateral acupuncture group (ipsi-group). The patients received one acupuncture treatment session at ST 38 on the nonpainful or painful sides, respectively. Before and after acupuncture treatment, they underwent functional magnetic resonance scanning. The treatment-related changes in degree centrality (DC) maps were compared between the two groups. We found alleviated pain and improved shoulder function in both groups, but better shoulder functional improvement was observed in the contra-group. Increased DC in the anterior/paracingulate cortex and decreased DC in bilateral postcentral gyri were found in the contra-group, while decreased DC in the bilateral cerebellum and right thalamus was observed in the ipsi-group. Furthermore, the DC value in the bilateral anterior/paracingulate cortex was positively correlated with the treatment-related change in the Constant-Murley score. The current study reveals different changes of DC patterns after acupuncture at contralateral or ipsilateral ST 38 in patients with CSP. Our findings support the hypothesis of acupoint specificity and provide the evidence for acupuncturists to select acupoints for CSP.

10.
Dalton Trans ; 2020 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-32412561

RESUMO

NbO-type MOFs built up from linear diisophthalate ligands and dicopper paddlewheel-based secondary building units offer an excellent platform to perform pore surface chemistry engineering and understand the structure-property relationship. In this work, we designed and synthesized two N-oxide functionalized linear diisophthalate ligands, and employed them to construct under suitable solvothermal conditions their corresponding NbO-type MOFs termed ZJNU-19 and ZJNU-20. Their gas adsorption properties with respect to C2H2, CO2, and CH4 were systematically measured, and adsorption selectivities of C2H2 and CO2 over CH4 were assayed using a well-known ideal adsorbed solution theory, establishing their promising potential for C2H2/CH4 and CO2/CH4 separations in connection with acetylene and natural gas separation and purification, which were found to be less dependent on the methyl position. In particular, at atmospheric pressure, the C2H2 and CO2 uptake capacities reach as high as 214.5 and 114.7 cm3 (STP) g-1 for ZJNU-19, and 210.0 and 111.9 cm3 (STP) g-1 for ZJNU-20 at 295 K, while the IAST-predicted C2H2/CH4 (v/v = 1/1) and CO2/CH4 (v/v = 1/1) adsorption selectivities are up to 42.2 and 6.4 for ZJNU-19, and 42.1 and 6.2 for ZJNU-20 at 298 K. Furthermore, a comparative investigation demonstrated that N-oxide is a powerful chemical functionality that can be utilized to design and construct porous framework compounds for boosting C2H2 and CO2 adsorptions.

11.
Int J Biol Macromol ; 158: 562-568, 2020 May 04.
Artigo em Inglês | MEDLINE | ID: mdl-32380112

RESUMO

Neonatal hypoxic-ischemic encephalopathy (HIE) is a complex condition that remains the leading cause of mortality and morbidity among infants. Polysaccharide has been reported to possess diverse biological activities, however, the neuro-protective activity of polysaccharide isolated from Lycium ruthenicum remains unknown so far. However, the role of Lycium ruthenicum polysaccharide 3 (LRP3) in HIE has not been evaluated. Herein, we investigated the effect of LRP3 on oxygen-glucose deprivation/reoxygenation (OGD/R)-induced primary cortical neurons. Our results demonstrated that LRP3 significantly improved the cell viability of OGD/R-induced cortical neurons. The OGD/R-caused increase in ROS production and decrease in the activities of anti-oxidative enzymes including catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) were mitigated by LRP3. Besides, the caspase-3 activity in OGD/R-induced cortical neurons was markedly decreased after LRP3 treatment. The increased bax expression and decreased bcl-2 expression caused by OGD/R stimulation were alleviated by pretreatment with LRP3. In addition, LRP3 significantly induced the expressions of nuclear factor erythroid 2-related factor (Nrf2) and heme oxygenase-1 (HO-1) in OGD/R-induced cortical neurons. However, inhibition of Nrf2/HO-1 signaling pathway through transfection with siRNA targeting Nrf2 reversed the protective effects of LRP3. In conclusion, LRP3 exerts a neuroprotective effect against OGD/R-induced neuronal injury in rat primary cortical neurons.

12.
J Ultrasound Med ; 2020 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-32383807

RESUMO

OBJECTIVES: To investigate whether quantitative contrast-enhanced ultrasound (CEUS) analysis and magnetic resonance imaging (MRI) can differentiate hepatic angiomyolipoma (AML) from hepatocellular carcinoma (HCC) and hepatic epithelial angiomyolipoma (EAML) from nonepithelial AML. METHODS: Retrospective analysis of 26 CEUS lesions pathologically confirmed as AML and 60 patients with confirmed HCC. Clinical and ultrasound (US) features of AML versus HCC and EAML versus nonepithelial AML were analyzed. Quantitative parameters, including arrival time, ascending slope, time to peak, peak intensity (PI), fall time (FT), and mean transit time (MTT), were analyzed. Diagnostic performance of MRI and CEUS was compared. RESULTS: Angiomyolipoma was significantly different from HCC in US findings of solitary tumor (P < .001), well-defined border (P = .025), and hyperechogenicity (P < .001). Tumor types were significantly different in US findings of centripetal filling pattern (P < .001), less hypoenhancement in the CEUS delayed phase (P = .04), high PI (P = .007) and ascending slope (P = .006), and short FT (P = .001) and MTT (P < .05). The sensitivity and specificity of quantitative parameters were 61.5% to 96.2% and 55.0% to 71.7% (area under the curve, 0.642-0.773). An early draining vein and tumor vessels, intratumor fat, absence of pseudocapsules, mild signal intensity on diffusion-weighted imaging, and hypointensity in the MRI delayed phase were helpful for differentiation of AML from HCC (P < .05). The positive predictive values of CEUS and MRI were 46.3% to 100% and 42.9% to 75%, with no significant difference (P > .05). Epithelial AML was more likely to be hypoechoic with a centripetal filling pattern than nonepithelial AML (P < .05). The MTT and FT of EAML were shorter than those of nonepithelial AML (P < .05). The PI of EAML was significantly stronger than that of nonepithelial AML (P < .01). The sensitivity and specificity of quantitative parameters were 68.7% to 100.0% and 60.0% to 100.0% (area under the curve, 0.825-0.941). Less intratumor fat was helpful for differentiation of EAML from nonepithelial AML (P < .05). The positive predictive values of CEUS and MRI were 77.8% to 100% and 85.7%, with a significant difference (P < .05). CONCLUSIONS: Quantitative CEUS findings could be valuable in differentiating EAML from HCC.

13.
Signal Transduct Target Ther ; 5(1): 60, 2020 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-32355263

RESUMO

Radiotherapy is one of the most common countermeasures for treating a wide range of tumors. However, the radioresistance of cancer cells is still a major limitation for radiotherapy applications. Efforts are continuously ongoing to explore sensitizing targets and develop radiosensitizers for improving the outcomes of radiotherapy. DNA double-strand breaks are the most lethal lesions induced by ionizing radiation and can trigger a series of cellular DNA damage responses (DDRs), including those helping cells recover from radiation injuries, such as the activation of DNA damage sensing and early transduction pathways, cell cycle arrest, and DNA repair. Obviously, these protective DDRs confer tumor radioresistance. Targeting DDR signaling pathways has become an attractive strategy for overcoming tumor radioresistance, and some important advances and breakthroughs have already been achieved in recent years. On the basis of comprehensively reviewing the DDR signal pathways, we provide an update on the novel and promising druggable targets emerging from DDR pathways that can be exploited for radiosensitization. We further discuss recent advances identified from preclinical studies, current clinical trials, and clinical application of chemical inhibitors targeting key DDR proteins, including DNA-PKcs (DNA-dependent protein kinase, catalytic subunit), ATM/ATR (ataxia-telangiectasia mutated and Rad3-related), the MRN (MRE11-RAD50-NBS1) complex, the PARP (poly[ADP-ribose] polymerase) family, MDC1, Wee1, LIG4 (ligase IV), CDK1, BRCA1 (BRCA1 C terminal), CHK1, and HIF-1 (hypoxia-inducible factor-1). Challenges for ionizing radiation-induced signal transduction and targeted therapy are also discussed based on recent achievements in the biological field of radiotherapy.

14.
Nanotechnology ; 2020 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-32375141

RESUMO

Conventional approaches for preparing yolk-shell nanostructures require complicated procedures such as multi-step coatings and template removal. Herein, we present a new and general strategy for making yolk-shell nanocomposites based on an oil-in-water emulsion system. As a demonstrating case, silicon nanoparticles were dispersed in an oil phase which was in an oil-in-water emulsion; then the oil/water interface was in-situ polymerized to form microcapsules. After carbonization, the shell of microcapsules was formed. The Li-ion battery anodes based on the microcapsules exhibit a good electrochemical performance including stable capacity and high rate-performance. The capacity remains 1100 mAh g-1 after 500 cycles at a current density of 1.9 A g-1, along with a Coulombic efficiency of ≈99.9%. In addition, the method presented here is general, which is applicable for the synthesis of many yolk shell-structured nanocomposites.

15.
JMIR Med Inform ; 8(5): e18251, 2020 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-32369039

RESUMO

BACKGROUND: Computer-aided diagnosis (CAD) is a tool that can help radiologists diagnose breast lesions by ultrasonography. Previous studies have demonstrated that CAD can help reduce the incidence of missed diagnoses by radiologists. However, the optimal method to apply CAD to breast lesions using diagnostic planes has not been assessed. OBJECTIVE: The aim of this study was to compare the performance of radiologists with different levels of experience when using CAD with the quadri-planes method to detect breast tumors. METHODS: From November 2018 to October 2019, we enrolled patients in the study who had a breast mass as their most prominent symptom. We assigned 2 ultrasound radiologists (with 1 and 5 years of experience, respectively) to read breast ultrasonography images without CAD and then to perform a second reading while applying CAD with the quadri-planes method. We then compared the diagnostic performance of the readers for the 2 readings (without and with CAD). The McNemar test for paired data was used for statistical analysis. RESULTS: A total of 331 patients were included in this study (mean age 43.88 years, range 17-70, SD 12.10), including 512 lesions (mean diameter 1.85 centimeters, SD 1.19; range 0.26-9.5); 200/512 (39.1%) were malignant, and 312/512 (60.9%) were benign. For CAD, the area under the receiver operating characteristic curve (AUC) improved significantly from 0.76 (95% CI 0.71-0.79) with the cross-planes method to 0.84 (95% CI 0.80-0.88; P<.001) with the quadri-planes method. For the novice reader, the AUC significantly improved from 0.73 (95% CI 0.69-0.78) for the without-CAD mode to 0.83 (95% CI 0.80-0.87; P<.001) for the combined-CAD mode with the quadri-planes method. For the experienced reader, the AUC improved from 0.85 (95% CI 0.81-0.88) to 0.87 (95% CI 0.84-0.91; P=.15). The kappa indicating consistency between the experienced reader and the novice reader for the combined-CAD mode was 0.63. For the novice reader, the sensitivity significantly improved from 60.0% for the without-CAD mode to 79.0% for the combined-CAD mode (P=.004). The specificity, negative predictive value, positive predictive value, and accuracy improved from 84.9% to 87.8% (P=.53), 76.8% to 86.7% (P=.07), 71.9% to 80.6% (P=.13), and 75.2% to 84.4% (P=.12), respectively. For the experienced reader, the sensitivity improved significantly from 76.0% for the without-CAD mode to 87.0% for the combined-CAD mode (P=.045). The NPV and accuracy moderately improved from 85.8% and 86.3% to 91.0% (P=.27) and 87.0% (P=.84), respectively. The specificity and positive predictive value decreased from 87.4% to 81.3% (P=.25) and from 87.2% to 93.0% (P=.16), respectively. CONCLUSIONS: S-Detect is a feasible diagnostic tool that can improve the sensitivity, accuracy, and AUC of the quadri-planes method for both novice and experienced readers while also improving the specificity for the novice reader. It demonstrates important application value in the clinical diagnosis of breast cancer. TRIAL REGISTRATION: ChiCTR.org.cn 1800019649; http://www.chictr.org.cn/showproj.aspx?proj=33094.

16.
Arch Biochem Biophys ; 686: 108351, 2020 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-32240636

RESUMO

Transforming growth factor beta regulator 4 (TBRG4) is a novel regulator in tumorigenic progression of several tumors. However, so far, the expression and functions of TBRG4 in osteosarcoma are unknown. The aim of this study was to investigate the potential biological functions of TBRG4 in osteosarcoma. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of TBRG4 in osteosarcoma tissues and cell lines. The levels of TBRG4 protein in osteosarcoma tissues were assessed by immunohistochemistry. Lentivirus-mediated short hairpin (sh) RNA was employed to knock down TBRG4 in osteosarcoma cells, and the expressions of TBRG4 mRNA and protein were determined by qRT-PCR and Western blot assay, respectively. Subsequently, the proliferation, clonogenic ability, apoptosis and invasion of osteosarcoma cells were measured using high content screening analysis and CCK8 assay, tumor sphere formation assay, flow cytometry and Transwell invasion assays, respectively. Furthermore, the osteosarcoma cells growth and metastasis in vivo were detected, and the effect of TBRG4 on the transforming growth factor ß1 (TGF-ß1) and PI3K/AKT signaling pathway was explored by qRT-PCR and Western blot assay, respectively. The results showed the levels of TBRG4 were overexpressed in osteosarcoma tissues and cell lines, confirming that the high TBRG4 expression was related to advanced tumor stages, large tumor size, and lymph node metastasis. Functional assays showed knockdown of TBRG4 could inhibit proliferation, invasion and induce apoptosis of osteosarcoma cells in vitro, and could also suppress osteosarcoma growth and metastasis in vivo. By examining the expression levels of TGF-ß1, p-PI3K, PI3K, p-AKT and AKT, it showed that the suppression of TBRG4 would reduce TGF-ß1 expression and inactivate the PI3K/AKT signaling pathway. These results showed for the first time that TBRG4 knockdown could suppress osteosarcoma progression, suggesting TBRG4 might be a promising therapeutic target for osteosarcoma treatment.

17.
Zhongguo Yi Liao Qi Xie Za Zhi ; 44(1): 33-37, 2020 Jan 08.
Artigo em Chinês | MEDLINE | ID: mdl-32343063

RESUMO

Accompanied by changes in modern work and lifestyle, the incidence of cervical spondylosis has increased year by year. In view of the fact long-term fixed posture of the head and neck is one of the main causes of cervical spondylosis, a set of wearable cervical spondylosis prevention system is developed. The system comprises a head and neck movement collection module based on the acceleration sensor and a head and neck motion recognition module based on artificial intelligence. Experimental results showed that the system can accurately identify long-term posture of the head and neck, and guide users to complete effective exercise therapy under the supervision of motion recognition module. Using this system can be beneficial for the prevention of cervical spondylosis.

18.
Artigo em Inglês | MEDLINE | ID: mdl-32314195

RESUMO

PURPOSE: Cystic fibrosis transmembrane conductance regulator (CFTR) and adhesion G protein-coupled receptor G2 (ADGRG2) have been identified as the main pathogenic genes in congenital bilateral absence of the vas deferens (CBAVD), which is an important cause of obstructive azoospermia. This study aimed to identify the disease-causing gene in two brothers with CBAVD from a Chinese consanguineous family and reveal the intracytoplasmic sperm injection (ICSI) outcomes in these patients. METHODS: Whole-exome sequencing and Sanger sequencing were used to identify the candidate pathogenic genes. Real-time polymerase chain reaction, immunohistochemistry, and immunofluorescence were used to assess the expression of the mutant gene. Moreover, the ICSI results from both patients were retrospectively reviewed. RESULTS: A novel hemizygous loss-of-function mutation (c.G118T: p.Glu40*) in ADGRG2 was identified in both patients with CBAVD. This mutation is absent from the human genome databases and causes an early translational termination in the third exon of ADGRG2. Expression analyses showed that both the ADGRG2 mRNA and the corresponding protein were undetectable in the proximal epididymal tissue of ADGRG2-mutated patients. ADGRG2 expression was restricted to the apical membranes of non-ciliated epithelia in human efferent ducts, which was consistent with a previous report in mice. Both ADGRG2-mutated patients had normal spermatogenesis and had successful clinical outcomes following ICSI. CONCLUSIONS: Our study verifies the pathogenic role of ADGRG2 in X-linked CBAVD and broadens the spectrum of ADGRG2 mutations. In addition, we found positive ICSI outcomes in the two ADGRG2-mutated CBAVD patients.

19.
Acupunct Med ; : 964528419900789, 2020 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-32309995

RESUMO

BACKGROUND: Acupuncture has been advocated for as a potentially effective therapy for patients with knee osteoarthritis (KOA) in systematic reviews and guidelines. However, there is still a lack of agreement on the optimal therapeutic protocol for acupuncture. This aim of this study was to develop an expert consensus regarding the therapeutic protocol of acupuncture to guide doctors in clinical practice. METHODS: An initial list of items was based on an overview of research evidence from four databases and clinical problem investigation with a multidisciplinary panel. A two-step process was used to optimize the list, including semi-structured interviews with three acupuncture clinical experts and a three-round Delphi consensus survey with the voting panel. A nine-point Likert-type scale (1 = strongly disagree, 9 = strongly agree) was used to measure agreement. RESULTS: In total, 52 professionals (response rate: 52%) confirmed their participation in the voting panel. The initial list including 28 items was evaluated. Following a three-round Delphi survey, a consensus was achieved including 37 items that can be broadly categorized into six domains: (1) main treatment principles, (2) acupuncture treatment, (3) dose of acupuncture intervention, (4) primary outcomes, (5) adverse events and (6) others. CONCLUSION: This expert consensus could be used to guide doctors in clinical practice and help patients with KOA gain access to appropriate and coordinated acupuncture treatment.

20.
Artigo em Inglês | MEDLINE | ID: mdl-32323121

RESUMO

BACKGROUND: Asthenoteratospermia with multiple morphological abnormalities in the sperm flagella (MMAF) is a significant cause of male infertility. WDR19 is a core component in the IFT-A complex and has a critical role in intraflagellar transport. However, the role of WDR19 mutations in male infertility has yet to be examined. METHODS AND RESULTS: We performed whole exome sequencing (WES) for 65 asthenoteratospermia individuals and identified a proband who carried a homozygous WDR19 (c.A3811G, p.K1271E) mutation from a consanguineous family. Systematic examinations, including CT scanning and retinal imaging, excluded previous ciliopathic syndromes in the proband. Moreover, semen analysis of this patient showed that the progressive rate decreased to zero, and the sperm flagella showed multiple morphological abnormalities. Scanning and transmission electron microscopy assays indicated that the ultrastructure of sperm flagella in the patient was completely destroyed, while immunofluorescence revealed that WDR19 was absent from the sperm neck and flagella. Moreover, IFT140 and IFT88, predicted to interact with WDR19 directly, were mis-allocated in the WDR19-mutated sperm. Notably, the MMAF subject harboring WDR19 variant and his partner successfully achieved clinical pregnancy through intracytoplasmic sperm injection (ICSI). CONCLUSIONS: We identified WDR19 as a novel pathogenic gene for male infertility caused by asthenoteratospermia in the absence of other ciliopathic phenotypes, and that patients carrying WDR19 variant can have favorable pregnancy outcomes following ICSI.

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