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1.
Cell Mol Life Sci ; 81(1): 108, 2024 Feb 29.
Artigo em Inglês | MEDLINE | ID: mdl-38421455

RESUMO

Spermiogenesis is considered to be crucial for the production of haploid spermatozoa with normal morphology, structure and function, but the mechanisms underlying this process remain largely unclear. Here, we demonstrate that SPEM family member 2 (Spem2), as a novel testis-enriched gene, is essential for spermiogenesis and male fertility. Spem2 is predominantly expressed in the haploid male germ cells and is highly conserved across mammals. Mice deficient for Spem2 develop male infertility associated with spermiogenesis impairment. Specifically, the insufficient sperm individualization, failure of excess cytoplasm shedding, and defects in acrosome formation are evident in Spem2-null sperm. Sperm counts and motility are also significantly reduced compared to controls. In vivo fertilization assays have shown that Spem2-null sperm are unable to fertilize oocytes, possibly due to their impaired ability to migrate from the uterus into the oviduct. However, the infertility of Spem2-/- males cannot be rescued by in vitro fertilization, suggesting that defective sperm-egg interaction may also be a contributing factor. Furthermore, SPEM2 is detected to interact with ZPBP, PRSS21, PRSS54, PRSS55, ADAM2 and ADAM3 and is also required for their processing and maturation in epididymal sperm. Our findings establish SPEM2 as an essential regulator of spermiogenesis and fertilization in mice, possibly in mammals including humans. Understanding the molecular role of SPEM2 could provide new insights into future therapeutic treatment of human male infertility and development of non-hormonal male contraceptives.


Assuntos
Infertilidade Masculina , Testículo , Humanos , Feminino , Masculino , Animais , Camundongos , Sêmen , Espermatogênese/genética , Infertilidade Masculina/genética , Interações Espermatozoide-Óvulo , Mamíferos , Fertilinas
2.
J Cell Sci ; 134(10)2021 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-34028541

RESUMO

A disintegrin and metalloproteinase 3 (ADAM3) is a sperm membrane protein critical for sperm migration from the uterus into the oviduct and sperm-egg binding in mice. Disruption of PRSS37 results in male infertility concurrent with the absence of mature ADAM3 from cauda epididymal sperm. However, how PRSS37 modulates ADAM3 maturation remains largely unclear. Here, we determine the PRSS37 interactome by GFP immunoprecipitation coupled with mass spectrometry in PRSS37-EGFP knock-in mice. Three molecular chaperones (CLGN, CALR3 and PDILT) and three ADAM proteins (ADAM2, ADAM6B and ADAM4) were identified to be interacting with PRSS37. Coincidently, five of them (except ADAM4) have been reported to interact with ADAM3 precursor and regulate its maturation. We further demonstrated that PRSS37 also interacts directly with ADAM3 precursor and its deficiency impedes the association between PDILT and ADAM3. This could contribute to improper translocation of ADAM3 to the germ cell surface, leading to ADAM3 loss in PRSS37-null mature sperm. The understanding of the maturation mechanisms of pivotal sperm plasma membrane proteins will pave the way toward novel strategies for contraception and the treatment of unexplained male infertility.


Assuntos
Infertilidade Masculina , Glicoproteínas de Membrana , Proteínas ADAM/genética , Animais , Epididimo , Feminino , Masculino , Glicoproteínas de Membrana/genética , Camundongos , Camundongos Knockout , Isomerases de Dissulfetos de Proteínas , Serina Proteases , Espermatozoides
3.
BMC Musculoskelet Disord ; 24(1): 941, 2023 Dec 05.
Artigo em Inglês | MEDLINE | ID: mdl-38053090

RESUMO

BACKGROUND: Cortical buttress are important factors for postoperative stable reconstruction of per/inter-trochanteric fractures. The study aimed to measure the remnant axial cortical length (RACL) of the proximal circumference of the femur, and to determine which part of the RACL can be used reliably to postoperatively sustain the head-neck fragment as a cortical support pattern. METHODS: Eighty patients with trochanteric hip fractures admitted from January 2015 to January 2016 were included in a retrospective study. Their pre-operative computed tomography (CT) images were used to form 3D-CT reconstructions via Mimics software. After simulated rotation and movement for fracture reduction, the RACL, its three component parts-namely, the remnant anterior cortex (RAC), remnant lateral cortex (RLC), and remnant posterior cortex (RPC) -the γ angle between the anterior and posterior cortex, and the Hsu's lateral wall thickness (LWT) were evaluated. RESULTS: Patients with an A1 fracture (21/80) had a longer RACL (88.8 ± 15.8 mm) than those with an A2 fracture (60.0 ± 11.9 mm; P < 0.01). The RAC, RLC, and RPC of the RACL in A1 fractures were also significantly longer than those in A2 fractures (P < 0.001). However, the most significant difference among the three components of the RACL was in the RPC, which was 27.3 ± 7.8 mm in A1 fractures and 9.2 ± 6.6 mm in A2 fractures. In addition, the coefficient of variation of the RAC was only 20.0%, while that of the RPC was 75.5%. The average γ angle in A1 fractures was 16.2 ± 13.1°, which was significantly smaller than that in A2 fractures, which was 40.3 ± 14.5° (P < 0.001). There was a significant statistical difference in the LWT between A1 and A2 fractures (P < 0.001). There were significant differences in the RACL, RAC, RLC, RPC, γ angle, and LWT among the five subtypes (P < 0.001). CONCLUSIONS: The RAC is relatively stable in pertrochanteric fractures. Fracture reduction through a RAC buttress may help to enhance the postoperative stable reconstruction of per/inter-trochanteric fractures and make possible good mechanical support for fracture healing.


Assuntos
Fraturas do Fêmur , Fraturas do Quadril , Humanos , Estudos Retrospectivos , Fraturas do Fêmur/cirurgia , Fêmur/cirurgia , Fraturas do Quadril/diagnóstico por imagem , Fraturas do Quadril/cirurgia , Tomografia Computadorizada por Raios X/métodos
4.
Biol Reprod ; 107(4): 1139-1154, 2022 10 11.
Artigo em Inglês | MEDLINE | ID: mdl-35863763

RESUMO

Serine proteases (PRSS) constitute nearly one-third of all proteases, and many of them have been identified to be testis-specific and play significant roles during sperm development and male reproduction. PRSS54 is one of the testis-specific PRSS in mouse and human but its physiological function remains largely unclear. In the present study, we demonstrate in detail that PRSS54 exists not only in testis but also in mature sperm, exhibiting a change in protein size from 50 kDa in testis to 42 kDa in sperm. Loss of PRSS54 in mice results in male subfertility, acrosome deformation, defective sperm-zona penetration, and phenotypes of male subfertility and acrosome deformation can be rescued by Prss54 transgene. Ultrastructure analyses by transmission electronic microscopy further reveal various morphological abnormalities of Prss54-/- spermatids during spermiogenesis, including unfused vacuoles in acrosome, detachment and eccentrical localization of the acrosomal granules, and asymmetrical elongation of the nucleus. Subcellular localization of PRSS54 display that it appears in the acrosomal granule at the early phase of acrosome biogenesis, then extends along the inner acrosomal membrane, and ultimately presents in the acrosome region of the mature sperm. PRSS54 interacts with acrosomal proteins ZPBP1, ZPBP2, ACRBP, and ZP3R, and loss of PRSS54 affects the distribution of these proteins in testis and sperm, although their protein levels are largely unaffected. Moreover, Prss54-/- sperm are more sensitive to acrosome reaction inducers.


Assuntos
Acrossomo , Infertilidade Masculina , Acrossomo/metabolismo , Animais , Proteínas de Transporte/metabolismo , Proteínas do Ovo , Humanos , Infertilidade Masculina/metabolismo , Masculino , Proteínas de Membrana/metabolismo , Camundongos , Morfogênese , Proteínas/metabolismo , Sêmen/metabolismo , Serina Endopeptidases/metabolismo , Serina Proteases/genética , Serina Proteases/metabolismo , Espermatozoides/metabolismo , Testículo/metabolismo
5.
BMC Musculoskelet Disord ; 23(1): 175, 2022 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-35209887

RESUMO

BACKGROUND: Maintaining anteromedial cortical support is essential for controlling sliding and decreasing postoperative implant-related complications. However, adequate fracture reduction with cortical support in immediate postoperative fluoroscopy is not invariable in postoperative follow-ups. This study was conducted to investigate the risk factors leading to anteromedial cortical support failure in follow up for pertrochanteric femur fractures treated with cephalomedullary nails. METHODS: This retrospective study enrolled 159 patients with pertrochanteric fractures (AO/OTA- 31A1 and 31A2) that fixed with cephalomedullary nails. All patients were evaluated as adequate fracture reduction in immediate postoperative fluoroscopy before leaving the operation theater. The patients were separated into two groups based on the condition of the anteromedial cortex in the postoperative 3D CT with full-range observation: those with calcar support maintained in Group 1 and those with calcar support lost in Group 2. Demographic information, fracture classification, TAD (tip-apex distance), Cal-TAD, Parker ratio, NSA (neck-shaft angle), reduction quality score, and calcar fracture gapping were collected and compared. Logistic regression analysis was conducted to explore the risk factors leading to anteromedial cortex change. RESULTS: Anteromedial cortical support failure was noted in 46 cases (29%). There was no significant difference between the two groups concerning age, sex, side injury, TAD, Cal-TAD, Parker ratio, or NSA. There was a significant difference in the AO/OTA fracture classification in univariate analysis but no difference in the multivariable analysis. The reduction quality score, calcar fracture gapping in the AP (anteroposterior), and lateral views were significantly associated with anteromedial cortical support failure in follow-up after cephalomedullary nailing in the multivariable analysis. The threshold value of calcar fracture gapping for the risk of loss was 4.2 mm in the AP and 3.8 mm in the lateral fluoroscopies. Mechanical complications (lateral sliding and varus) were frequently observed in the negative anteromedial cortical support group. CONCLUSIONS: Good reduction quality was a protective factor, and larger calcar fracture gapping in the AP and lateral views were risk factors leading to the postoperative loss of anteromedial cortical support. Therefore, we should pay close attention to fracture reduction and minimize the calcar fracture gap during surgery.


Assuntos
Fixação Intramedular de Fraturas , Fraturas do Quadril , Pinos Ortopédicos/efeitos adversos , Fêmur/cirurgia , Fixação Intramedular de Fraturas/efeitos adversos , Fraturas do Quadril/diagnóstico por imagem , Fraturas do Quadril/etiologia , Fraturas do Quadril/cirurgia , Humanos , Estudos Retrospectivos , Resultado do Tratamento
6.
Biol Reprod ; 105(4): 789-807, 2021 10 11.
Artigo em Inglês | MEDLINE | ID: mdl-34131698

RESUMO

Sexual reproduction requires the fusion of two gametes in a multistep and multifactorial process termed fertilization. One of the main steps that ensures successful fertilization is acrosome reaction. The acrosome, a special kind of organelle with a cap-like structure that covers the anterior portion of sperm head, plays a key role in the process. Acrosome biogenesis begins with the initial stage of spermatid development, and it is typically divided into four successive phases: the Golgi phase, cap phase, acrosome phase, and maturation phase. The run smoothly of above processes needs an active and specific coordination between the all kinds of organelles (endoplasmic reticulum, trans-Golgi network, and nucleus) and cytoplasmic structures (acroplaxome and manchette). During the past two decades, an increasing number of genes have been discovered to be involved in modulating acrosome formation. Most of these proteins interact with each other and show a complicated molecular regulatory mechanism to facilitate the occurrence of this event. This review focuses on the progresses of studying acrosome biogenesis using gene-manipulated mice and highlights an emerging molecular basis of mammalian acrosome formation.


Assuntos
Acrossomo/fisiologia , Espermatogênese , Animais , Masculino , Camundongos , Camundongos Transgênicos
7.
J Reprod Dev ; 66(1): 57-65, 2020 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-31801914

RESUMO

Testis-specific genes are prone to affect spermatogenesis or sperm fertility, and thus may play pivotal roles in male reproduction. However, whether a gene really affects male reproduction in vivo needs to be confirmed using a gene knock-out (KO) model, a 'gold standard' method. Increasing studies have found that some of the evolutionarily conserved testis-enriched genes are not essential for male fertility. In this study, we report that 1700121C10Rik, a previously uncharacterized gene, is specifically expressed in the testis and produces two long noncoding RNAs (lncRNAs) in mouse: Transcript 1 and Transcript 2. qRT-PCR, northern blotting, and in situ hybridization revealed that expression of both the lncRNAs commenced at the onset of sexual maturity and was predominant in round and elongating spermatids during spermiogenesis. Moreover, we found different subcellular localization of Transcript 1 and Transcript 2 that was predominant in the cytoplasm and nucleus, respectively. 1700121C10Rik-KO mouse model disrupting Transcript 1 and Transcript 2 expression was generated by CRISPR/Cas9 to determine their role in male reproduction. Results showed that 1700121C10Rik-KO male mice were fully fertile with approximately standard testis size, testicular histology, sperm production, sperm morphology, sperm motility, and induction of acrosome reaction. Thus, we conclude that both the testis-specific 1700121C10Rik-produced lncRNAs are dispensable for male fertility in mice under standard laboratory conditions.


Assuntos
Fertilidade/genética , Infertilidade Masculina/genética , RNA Longo não Codificante/genética , Espermatogênese/genética , Testículo/metabolismo , Animais , Infertilidade Masculina/metabolismo , Masculino , Camundongos , Camundongos Knockout , RNA Longo não Codificante/metabolismo , Motilidade dos Espermatozoides/genética
8.
BMC Musculoskelet Disord ; 21(1): 719, 2020 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-33153440

RESUMO

BACKGROUND: Anteromedial cortex-to-cortex reduction is a key parameter for stable reconstruction of the fracture fragments during the intertrochanteric fracture fixation. This paper introduces the oblique fluoroscopic projection as a novel method to evaluate the quality of anteromedial cortical apposition. METHODS: Three proximal femur specimens were marked with steel wires along five anatomic landmarks: Greater trochanter, Lesser trochanter, Intertrochanteric line, Anterolateral tubercle and the Anteromedial cortical line. After obtaining the standard femoral neck AP and lateral fluoroscopic images, the C-arm was rotated by every 5°increments until a clear tangential view of the antero-medial-inferior corner cortex was observed. 98 cases of intertrochanteric hip fractures were enrolled from April 2018 to October 2019. After fixation with the nails, the intra-operative anteromedial cortex reduction quality was evaluated from the AP, the true lateral, and the new anteromedial oblique fluoroscopic images. The fluoroscopic results were compared with the post-operative 3D-CT reconstruction images. RESULTS: The specimen study showed that internal rotation of the C-arm to approximately 30 ° can remove all the obscure shadows and clearly display the antero-medial-inferior cortical tangent line. Clinically,the positive, neutral and negative apposition of different cortices via intra-operative fluoroscopic images showed79, 19 and 0 cases of medial cortical apposition in AP views; 2, 68 and 28 cases of anterior cortices in lateral views;and 22, 51 and 25cases of anteromedial cortical apposition in oblique views respectively. The post-operative 3D-CT reconstruction images revealed that the final anteromedial cortical contact was noted in 62 cases (63.3%), and lost in 36 cases (36.7%). The overall coincidence rate between intra-operative fluoroscopy and post-operative 3D-CT was 63.3% (62/98) in AP view,79.6% (78/98) in lateral view, and 86.7% (85/98) in oblique view(p < 0.001). Negative cortical apposition in oblique view was highly predictive of a final loss of cortical support on 3D CT (24/25 cases, 96%).And non-negative cortical apposition in oblique view was highly associated with true cortical support on 3D CT images (61/73 cases, 83.6%) (p < 0.001). CONCLUSIONS: Besides the AP and lateral projections, an anteromedial oblique view of 30° certifies to be a very useful means for evaluation of the fracture reduction quality of anteromedial cortical apposition.


Assuntos
Fixação Intramedular de Fraturas , Fraturas do Quadril , Fêmur/diagnóstico por imagem , Fêmur/cirurgia , Fluoroscopia , Fixação de Fratura , Fixação Interna de Fraturas , Fraturas do Quadril/diagnóstico por imagem , Fraturas do Quadril/cirurgia , Humanos
9.
Mol Reprod Dev ; 86(8): 935-955, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31131960

RESUMO

Mammalian reproductive processes involve spermatogenesis, which occurs in the testis, and fertilization, which takes place in the female genital tract. Fertilization is a successive, multistep, and extremely complicated event that usually includes sperm survival in the uterus, sperm migration through the uterotubal junction (UTJ) and the oviduct, sperm penetration through the cumulus cell layer and the zona pellucida, and sperm-egg fusion. There may be a complex molecular mechanism to ensure that the above processes run smoothly. Previous studies have discovered essential factors for these fertilization steps through in vitro fertilization experiments. However, recent gene disruption approaches in mice have revealed that many of the factors previously described as important for fertilization are largely dispensable in gene-knockout animals, and some previously unknown factors are emerging. As a result, the molecular mechanisms of fertilization, especially sperm migration from the uterus into the oviduct, have recently been revised by the emergence of genetically modified animals. In this review, we only focus on and update the essential genes for sperm migration through the UTJ and describe recent advances in our knowledge of the basis of mammalian sperm migration.


Assuntos
Oviductos/metabolismo , Motilidade dos Espermatozoides , Espermatozoides/metabolismo , Útero/metabolismo , Animais , Feminino , Masculino , Camundongos , Camundongos Knockout , Oviductos/citologia , Espermatozoides/citologia , Útero/citologia
10.
Med Sci Monit ; 25: 2049-2057, 2019 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-30889172

RESUMO

BACKGROUND The lesser trochanter (LT) posterior cortical extension (LTPE) fragment is important for joint stability during reconstruction in unstable pertrochanteric hip fractures. This study aimed to investigate the morphological characteristics of the LT fragments using three-dimensional computed tomography (3-D CT) reconstruction and to compare clinical outcomes in different subgroups of 31A2 fractures. MATERIAL AND METHODS A retrospective study included CT images of 58 cases of 31A2 type unstable pertrochanteric fractures, using the Arbeitsgemeinschaft für Osteosynthesefragen and Orthopedic Trauma Association (AO/OTA) classification. After 3-D CT reconstruction, all the displaced fragments were reduced and the morphologic parameters of the LT fragments were measured. RESULTS At the mid-level of the LT, the mean cortical extension of the LT fragment was 33.5 mm in the posterior wall (83%), 19.0 mm in the medial wall (53%). The mean distal cortical extension from the lower edge of the LT was 13.0 mm. The LT fragment occupied 38% of the whole cortical circumference. Comparison of the subgroups showed that the LT fragment of A2.3 had a larger cortical extension in the posterior, medial, and distal cortex. The mean fracture collapse was 4.7 mm in A2.2 fractures and 5.4 mm in A2.3 fractures (p=0.311). Despite the increased size of the LT fragment in subtype A2.3 fracture compared with subtype A2.2 fracture, this did not influence the clinical outcome. CONCLUSIONS The 3-D morphology of the LT fragment from CT imaging provided a better understanding of the characteristics of subgroups of unstable pertrochanteric fractures, which may improve and guide implant choice.


Assuntos
Fêmur/diagnóstico por imagem , Fraturas do Quadril/diagnóstico por imagem , Tomografia Computadorizada por Raios X/métodos , Idoso , Idoso de 80 Anos ou mais , Parafusos Ósseos , Feminino , Fraturas do Fêmur/cirurgia , Fêmur/patologia , Fêmur/cirurgia , Fixação Interna de Fraturas/métodos , Fraturas do Quadril/patologia , Fraturas do Quadril/cirurgia , Humanos , Imageamento Tridimensional/métodos , Masculino , Estudos Retrospectivos
11.
Pestic Biochem Physiol ; 159: 107-117, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31400772

RESUMO

Latrophilin (LPH) is an adhesion G protein-coupled receptor (aGPCR) that participates in multiple essential physiological processes. Our previous studies have shown that lph is not only indispensable for the development and reproduction of red flour beetles (Tribolium castaneum), but also for their resistance against dichlorvos or carbofuran insecticides. However, the regulatory mechanism of lph-mediated insecticide susceptibility remains unclear. Here, we revealed that knockdown of lph in beetles resulted in opposing changes in two chemoreception genes, chemosensory protein 10 (CSP10) and odorant-binding protein C01 (OBPC01), in which the expression of TcCSP10 was downregulated, whereas the expression of TcOBPC01 was upregulated. TcCSP10 and TcOBPC01 were expressed at the highest levels in early pupal and late larval stages, respectively. High levels of expression of both these genes were observed in the heads (without antennae) of adults. TcCSP10 and TcOBPC01 were significantly induced by dichlorvos or carbofuran between 12 and 72 h (hrs) after exposure, suggesting that they are likely associated with increasing the binding affinity of insecticides, leading to a decrease in sensitivity to the insecticides. Moreover, once these two genes were knocked down, the susceptibility of the beetles to dichlorvos or carbofuran was enhanced. Additionally, RNA interference (RNAi) targeting of lph followed by exposure to dichlorvos or carbofuran also caused the opposing expression levels of TcCSP10 and TcOBPC01 compared to the expression levels of wild-type larvae treated with insecticides alone. All these results indicate that lph is involved in insecticide susceptibility through positively regulating TcCSP10; and the susceptibility could also further partially compensated for through the negative regulation of TcOBPC01 when lph was knockdown in the red flour beetle. Our studies shed new light on the molecular regulatory mechanisms of lph related to insecticide susceptibility.


Assuntos
Proteínas de Insetos/metabolismo , Inseticidas/farmacologia , Receptores de Peptídeos/metabolismo , Tribolium/efeitos dos fármacos , Tribolium/metabolismo , Animais , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Regulação da Expressão Gênica no Desenvolvimento/genética , Proteínas de Insetos/genética
12.
Int Heart J ; 60(1): 181-184, 2019 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-30464117
13.
Genetica ; 146(3): 287-302, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29797154

RESUMO

Latrophilin of Tribolium castaneum (Tclph) has been reported to play crucial roles in growth, development and reproduction. However, the regulatory mechanism of Tclph associated with these physiology processes is unknown. Thus, the global transcriptome profiles between RNAi treated (ds-Tclph) and control larvae of T. castaneum were analyzed by RNA-sequencing. Totally, 274 differentially expressed genes (DEGs) were identified between the ds-Tclph and control samples. These DEGs were classified into 42 GO functional groups, including developmental process, reproduction and stress response. The results indicated that knockdown of Tclph disturbed the antioxidant activity process, and partially inhibited the serine protease (SP) and lipase signaling pathways to regulate the development and reproduction as well as the decreasing of the stress response in T. castaneum. Additionally, knockdown of Tclph suppressed IMD immunity pathways which likely modulated the effects of Tclph on stress response. Interestingly, CSPs, ESTs, CYPs, AOXs and BGs were significantly down-regulated in ds-Tclph larvae, implying that they cooperated with Tclph to reduce the activity of cellular metabolism system. FMOs was up-regulated in ds-Tclph insects suggested it may be involved in detoxifying alkaloid of insect metabolism system. These results implied that Tclph participated in phase 0, I and II cellular detoxification. Furthermore, RNAi against Tclph increased larval susceptibility to carbamates and organophosphates insecticides, supporting that Tclph was indeed involved into the insecticide susceptibility in T. castaneum.


Assuntos
Resistência a Medicamentos , Regulação da Expressão Gênica/efeitos dos fármacos , Inseticidas/farmacologia , Receptores de Peptídeos/genética , Reprodução/genética , Transcriptoma , Tribolium/efeitos dos fármacos , Tribolium/fisiologia , Animais , Biologia Computacional/métodos , Perfilação da Expressão Gênica , Ontologia Genética , Genoma de Inseto , Genômica/métodos , Sequenciamento de Nucleotídeos em Larga Escala , Anotação de Sequência Molecular , Interferência de RNA
14.
Genomics ; 109(5-6): 362-373, 2017 10.
Artigo em Inglês | MEDLINE | ID: mdl-28624536

RESUMO

To date, although some microRNAs (miRNAs) have been discovered in the holometabolism insect Tribolium castaneum, large numbers of miRNAs still require investigation. Knocking down Dicer-1 (Dcr-1) and Argonaute-1 (Ago-1) in late larvae impaired miRNA synthesis, affected the juvenile hormone pathway by up-regulating Methoprene-tolerant (Met) and Krüppel-homolog1 (Kr-h1) transcript levels, and resulted in a series of defects in T. castaneum development and metamorphosis. Thus, high-throughput Illumina/Solexa sequencing was performed with a mixed sample of eight key developmental stages of T. castaneum. In total, 1154 unique miRNAs were discovered containing 274 conserved miRNAs belong to 68 miRNA families, 108 known candidate miRNAs and 772 novel miRNAs. Genome locus analysis showed that miRNA clusters are more abundant in T. castaneum than other species. The results indicated that RNAi of Dcr-1 and Ago-1 in T. castaneum resulted in miRNA-induced metamorphosis defects. Furthermore, large numbers of novel miRNAs were discovered in T. castaneum and localized to T. castaneum genome loci.


Assuntos
Proteínas Argonautas/genética , MicroRNAs/genética , RNA Helicases/genética , Fatores de Transcrição/genética , Tribolium/fisiologia , Animais , Análise por Conglomerados , Evolução Molecular , Regulação da Expressão Gênica no Desenvolvimento , Técnicas de Silenciamento de Genes , Sequenciamento de Nucleotídeos em Larga Escala , Proteínas de Insetos/genética , Fatores de Transcrição Kruppel-Like/genética , Metamorfose Biológica , Análise de Sequência de RNA , Tribolium/genética
15.
Int Orthop ; 42(1): 183-189, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-28891021

RESUMO

PURPOSE: Anteromedial cortical support reduction is favourable for secondary stability after limited sliding in unstable pertrochanteric fractures. The aim of this study was to compare the accuracy and agreement between intra-operative fluoroscopy and post-operative 3D reconstruction. MATERIALS AND METHODS: A retrospective analysis of 28 patients (mean 81.6 years) treated with short cephalomedullary nails was performed. All patients had full sets of intra-operative fluoroscopy and post-operative 3D CT images. Observation was focused on the position of the anteromedial cortices of the inferior corner between the head-neck fragment and femoral shaft, and their relationship was categorised into three types: positive, neutral and negative. The percentage of subsequent changes in cortical reduction quality between fluoroscopy and 3D CT was calculated and compared. RESULTS: There were 24 positive (85.7%), four neutral and no negative positions in the anteroposterior (AP) view and one positive, 20 neutral (71.4%) and seven negative positions in the lateral view from fluoroscopy. On post-operative 3D CT images with a full range of rotation, definitive anteromedial cortical contact (positive and neutral support) was observed in 18 cases (64.3%). Ten cases lost the anteromedial cortical buttress. With the posteromedial region of the lesser trochanter detached, a positive AP cortical position combined with a positive/neutral lateral position on fluoroscopy (17 cases) was highly predictive of reliable, definitive cortical support (15 cases, 88.2%) on 3D CT. A negative lateral position on fluoroscopy (seven cases), regardless of the combination in the AP view, was likely to predict the final loss of cortical support (six cases, 85.7%) on 3D CT; a positive/neutral lateral position (21 cases) was only associated with loss of support in four cases (19.1%) (p < 0.05). CONCLUSIONS: A lateral negative position of the anterior cortex on fluoroscopy for unstable pertrochanteric fractures may be highly predictive of post-operative final loss of the anteromedial cortical buttress, which should be avoided during operation.


Assuntos
Fraturas do Fêmur/diagnóstico por imagem , Fluoroscopia/métodos , Fixação Interna de Fraturas/métodos , Imageamento Tridimensional/métodos , Tomografia Computadorizada por Raios X/métodos , Adulto , Idoso , Pinos Ortopédicos/efeitos adversos , Feminino , Fraturas do Fêmur/cirurgia , Fêmur/cirurgia , Fixação de Fratura/métodos , Humanos , Masculino , Pessoa de Meia-Idade , Período Pós-Operatório , Estudos Retrospectivos
16.
Genesis ; 55(12)2017 12.
Artigo em Inglês | MEDLINE | ID: mdl-29052315

RESUMO

Latrophilins (LPHs) are adhesion G-protein-coupled receptors comprising three paralogous forms (LPH-1, LPH-2, and LPH-3) and known receptors for α-latrotoxin, which are involved in growth, development, adaptability, and schizophrenia and other diseases in vertebrates. However, the functions of LPH are poorly understood in most insects. Here, phylogenetic and synteny analysis indicated that LPH-1 and LPH-3 evolved separately from a common ancestor LPH-2. Then, latrophilin (Tclph) was cloned in Tribolium castaneum, and three alternatively spliced transcripts (Tclpha, Tclphb and Tclphc) were identified. All these three Tclphs were highest expressed at the early adult stage, and strongly expressed in central nervous system of adults. Larval RNA interference (RNAi) against Tclph caused 24% adult wing abnormal, 30% insect death, and led to 100% reductions in beetle fecundity. Fecundity deficiency was rescued by reciprocal crosses with wild-type females, but not males. And dissection results revealed that 63% of dsTclph female ovaries were atrophied. Further, exon-specific RNAi illustrated that neither knockdown of Tclpha nor Tclphc resulted in development defects and reductions in beetle fecundity. Thus, it indicated that Tclphb was essential for development and female fecundity in T. castaneum. Moreover, Tclph knockdown increased the expression of the foxo, plc, and pka genes, which most likely modulated the effects of Tclph on development and reproduction in T. castaneum.


Assuntos
Evolução Molecular , Fertilidade/genética , Receptores de Peptídeos/genética , Tribolium/genética , Processamento Alternativo/genética , Animais , Sistema Nervoso Central/crescimento & desenvolvimento , Sistema Nervoso Central/metabolismo , Embrião não Mamífero , Desenvolvimento Embrionário , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Larva/genética , Larva/crescimento & desenvolvimento , Filogenia , Isoformas de Proteínas/genética , Receptores Acoplados a Proteínas G/genética , Tribolium/crescimento & desenvolvimento
17.
J Ultrasound Med ; 36(3): 557-563, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28127781

RESUMO

OBJECTIVES: Virtual Touch tissue imaging quantification (VTIQ; Siemens Medical Solutions, Mountain View, CA) is useful for assessing tissue hardness. This study aimed to investigate the value of VTIQ in differential diagnosis of cervical lymph nodes. METHODS: We retrospectively analyzed conventional sonograms and VTIQ images of 85 pathologically confirmed patients with superficial lymph node lesions. Conventional sonography was first performed, with 2-dimensional images described. Then VTIQ shear wave velocity (SWV) values of superficial lymph nodes were measured. With pathologic diagnosis as the reference standard, a receiver operating characteristic curve was generated to evaluate VTIQ efficacy in differential diagnosis of metastatic and nonmetastatic cervical lymph nodes. RESULTS: Of the 85 nodes, 44 and 41 were metastatic and nonmetastatic, respectively. The latter group included 24 and 17 hematologic/lymphatic system disease and reactive hyperplastic nodes, respectively. Shear wave velocity values of metastatic nodes were significantly higher than those of their nonmetastatic counterparts (P < .001). With an area under the curve (AUC) of 0.953 and SWV cutoff of 3.27 m/s, accuracy, sensitivity, and specificity were 89.4%, 88.6%, and 90.2%, respectively, for distinguishing metastatic and nonmetastatic nodes. An AUC of 0.943 and SWV cutoff of 3.23 m/s yielded accuracy, sensitivity, and specificity of 88.2%, 88.6%, and 87.5% for differentiating metastatic from hematologic/lymphatic system disease nodes. Finally, an AUC of 0.968 and SWV cutoff of 3.27 m/s yielded accuracy, sensitivity, and specificity of 90.2%, 88.6%, and 94.1% for differentiating metastatic from reactive hyperplastic nodes. CONCLUSIONS: Virtual Touch tissue imaging quantification is efficient in differential diagnosis of metastatic and nonmetastatic cervical lymph nodes.


Assuntos
Processamento de Imagem Assistida por Computador/métodos , Linfonodos/diagnóstico por imagem , Doenças Linfáticas/diagnóstico por imagem , Ultrassonografia/métodos , Interface Usuário-Computador , Adulto , Idoso , Idoso de 80 Anos ou mais , Diagnóstico Diferencial , Feminino , Humanos , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Pescoço , Curva ROC , Reprodutibilidade dos Testes , Estudos Retrospectivos , Sensibilidade e Especificidade
18.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi ; 38(3): 290-297, 2024 Mar 15.
Artigo em Chinês | MEDLINE | ID: mdl-38500421

RESUMO

Objective: To analyze the causes and treatment of off target of the distal interlocking screws when short cephalomeduallry nails were installed through jig-guided targeting device, and to put forward the technical points to prevent off target. Methods: Retrospective analysis of 9 patients with intertrochanteric fractures treated between July 2014 and June 2023 was conducted, in which off target occurred during the insertion of the distal interlocking screw by jig-guided targeting device in short cephalomedullary nailling (<24 cm). There were 1 male and 8 females, with an average age of 82.7 years (range, 73-94 years). There were 3 cases of type A1, 5 cases of type A2, and 1 case of type A3 according to 2018-AO/Orthopaedic Trauma Association (AO/OTA) fracture classification. As for the misaligned distal interlocking screw, six parameters were collected and analyzed, including the time of finding, the position, the type of passing through the cortical bone, the special circumstances during operation (such as the need to remove the intramedullary nail for reaming the diaphysis, hammering, etc.), the treatment, and the patient follow-up results. Results: In the 9 patients, the off target of the distal interlocking screw was found in 7 cases during operation and in 2 cases after operation; the locking screw was located behind the nail in 7 cases and in front of the nail in 2 cases; the off target locking screw was passing tangentially in transcortical patern in 6 cases and in bicortical pattern through the medullary cavity in 3 cases. Three cases were attributed to the mismatch between the nail and the femur, two of which were attributed to the narrow femoral medullary cavity, one of which was attributed to the large anterolateral femoral bowing, and the other 6 cases were attributed to technical errors such as the loosening of the jig-guided targeting device, the tension of the fascia lata, and the blunt of the drill. In the 7 cases found during operation, the misaligned interlocking screw was removed first and the screw hole was left vacant, then in 2 cases, the interlocking screw was not used further; in 1 case, the distal dynamic hole was successfully inserted with a dynamic guide frame, and in 4 cases, the interlocking screw was successfully put after 2-3 attempts, leaving a large hole in the lateral cortex. No special treatment was performed in 2 cases found after operation. One patient was out of bed early after operation, 7 patients were in bed for 1 month, and 1 patient deteriorated to A3 type after operation and was in bed for 3 months. All the 9 patients were followed up 6-12 months, with an average of 8 months. Fracture healing was achieved in 8 patients. One patient with vacant interlocking screw had a secondary spiral fracture of the femoral shaft 3 months later, and was refixed with a long cephalomedullary nail and circlage wiring. Conclusion: Distal interlocking screw off target is rare, but when it occurs, it leaves a large cortical hole in the osteoporotic femoral shaft, reducing bone strength; the use of precision instruments and attention to technical details can reduce this adverse phenomenon.


Assuntos
Fraturas do Fêmur , Fixação Intramedular de Fraturas , Fraturas do Quadril , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pinos Ortopédicos , Parafusos Ósseos , Fraturas do Fêmur/cirurgia , Fêmur , Fixação Intramedular de Fraturas/métodos , Fraturas do Quadril/cirurgia , Fraturas do Quadril/etiologia , Estudos Retrospectivos , Resultado do Tratamento , Idoso
19.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi ; 38(1): 107-112, 2024 Jan 15.
Artigo em Chinês | MEDLINE | ID: mdl-38225849

RESUMO

Objective: To review the advancement made in the understanding of valgus impacted proximal humeral fracture (PHF). Methods: The domestic and foreign literature about the valgus impacted PHF was extensively reviewed and the definition, classification, pathological features, and treatment of valgus impacted PHFs were summarized. Results: PHF with a neck shaft angle ≥160° is recognized as a valgus impacted PHF characterized by the preservation of the medial epiphyseal region of the humeral head, which contributes to maintenance of the medial periosteum's integrity after fracture and reduces the occurrence of avascular necrosis. Therefore, the valgus impacted PHF has a better prognosis when compared to other complex PHFs. The Neer classification designates it as a three- or four-part fracture, while the AO/Association for the Study of Internal Fixation (AO/ASIF) categorizes it as type C (C1.1). In the management of the valgus impacted PHF, the selection between conservative and surgical approaches is contingent upon the patient's age and the extent of fracture displacement. While conservative treatment offers the advantage of being non-invasive, it is accompanied by limitations such as the inability to achieve anatomical reduction and the potential for multiple complications. Surgical treatment includes open reduction combined with steel wire or locking plate and/or non-absorbable suture, transosseous suture technology, and shoulder replacement. Surgeons must adopt personalized treatment strategies for each patient with a valgus impacted PHF. Minimally invasive surgery helps to preserve blood supply to the humeral head, mitigate the likelihood of avascular necrosis, and reduce postoperative complications of bone and soft tissue. For elderly patients with severe comminuted and displaced fractures, osteoporosis, and unsuitable internal fixation, shoulder joint replacement is the best treatment option. Conclusion: Currently, there has been some advancement in the classification, vascular supply, and management of valgus impacted PHF. Nevertheless, further research is imperative to assess the clinical safety, biomechanical stability, and indication of minimally invasive technology.


Assuntos
Fraturas Cominutivas , Fraturas do Úmero , Osteonecrose , Fraturas do Ombro , Idoso , Humanos , Placas Ósseas , Fios Ortopédicos , Fixação Interna de Fraturas/efeitos adversos , Fraturas Cominutivas/cirurgia , Estudos Retrospectivos , Fraturas do Ombro/cirurgia , Resultado do Tratamento
20.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi ; 38(4): 391-397, 2024 Apr 15.
Artigo em Chinês | MEDLINE | ID: mdl-38632056

RESUMO

Objective: To investigate the position of the anterior fracture line in AO/Orthopaedic Trauma Association (AO/OTA) type A2 unstable intertrochanteric fractures and its impact on the incidence of anterior cortical reduction loss after cephalomedullary nail fixation. Methods: A clinical data of 95 patients with intertrochanteric fractures who met the selection criteria between April 2020 and February 2023 was retrospectively analyzed. All patients were treated with cephalomedullary nail fixation, and the intra- and post-operative imaging data were complete. Among them, there were 37 males and 58 females. The age ranged from 61 to 97 years, with an average of 79.6 years. The time from injury to operation ranged from 7 hours to 11 days, with an average of 2.8 days. According to the 2018-AO/OTA classification standard, there were 39 cases of type 31-A2.2 and 56 cases of type 31-A2.3. Intraoperative fluoroscopy was used to record the number of patients with satisfactory fracture alignment. The preoperative CT data were imported into Mimics17.0 software to simulate the fracture reduction and measure the distance between the anterior fracture line and the intertrochanteric line bony ridge. The fractures were classified as transcapsular fractures, extra-capsular fractures, and intra-capsular fractures according to the distance. CT three-dimensional reconstruction was performed within 2 weeks after operation to observe the number of patients with anterior cortical reduction loss. The postoperative anterior cortical reduction loss incidence in patients with satisfactory fracture alignment, and the relationship between postoperative anterior cortical reduction loss and the position of the anterior fracture line were observed. Results: There were 52 cases (54.7%) of transcapsular fractures, 24 cases (25.3%) of extra-capsular fractures, and 19 cases (20.0%) of intra-capsular fractures. Among them, 41 of the 52 transcapsular fractures had satisfactory fracture alignment, and 4 (9.8%) of them experienced anterior cortical reduction loss after operation; 19 of the 24 extra-capsular fractures had satisfactory fracture alignment, and no anterior cortical reduction loss occurred; 16 of the 19 intra-capsular fractures had satisfactory fracture alignment, and 7 (43.8%) of them experienced anterior cortical reduction loss after operation. There was a significant difference in the incidence of anterior cortical reduction loss between groups ( χ 2=8.538, P=0.003). All patients were followed up 3-26 months (mean, 9 months). Among them, 91 cases had fracture healing, and 4 cases had nonunion. Conclusion: In AO/OTA type A2 unstable intertrochanteric fractures, where the anterior fracture line is located within the joint capsule, there is a high risk of anterior cortical reduction loss after operation.


Assuntos
Fixação Intramedular de Fraturas , Fraturas do Quadril , Masculino , Feminino , Humanos , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Estudos Retrospectivos , Pinos Ortopédicos , Resultado do Tratamento , Fraturas do Quadril/cirurgia
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