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Isodesmic reactions, in which chemical bonds are redistributed between substrates and products, provide a general and powerful strategy for both biological and chemical synthesis. However, most isodesmic reactions involve either metathesis or functional-group transfer. Here, we serendipitously discovered a novel isodesmic reaction of indoles and anilines that proceeds intramolecularly under weakly acidic conditions. In this process, the five-membered ring of the indole motif is broken and a new indole motif is constructed on the aniline side, accompanied by the formation of a new aniline motif. Mechanistic studies revealed the pivotal role of σâπ* hyperconjugation on the nitrogen atom of the indole motif in driving this unusual isodesmic reaction. Furthermore, we successfully synthesized a diverse series of polycyclic indole derivatives; among quinolines, potential antitumor agents were identified using cellular and in vivo experiments, thereby demonstrating the synthetic utility of the developed methodology.
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The process of generating type I/II collagen scaffolds is fraught with bubble formation, which can interfere with the three-dimensional structure of the scaffold. Herein, we applied low-temperature vacuum freeze-drying to remove mixed air bubbles under negative pressure. Type I and II rubber sponges were acid-solubilized via acid lysis and enzymolysis. Thereafter, vacuum negative pressure was applied to remove bubbles, and the cover glass press method was applied to shape the type I/II original scaffold. Vacuum negative pressure was applied for a second time to remove any residual bubbles. Subsequent application of carbamide/N-hydroxysuccinimide cross-linked the scaffold. The traditional method was used as the control group. The structure and number of residual bubbles and pore sizes of the two scaffolds were compared. Based on the relationship between the pressure and the number of residual bubbles, a curve was created, and the time of ice formation was calculated. The bubble content of the experimental group was significantly lower than that of the control group (P < 0.05). The pore diameter of the type I/II collagen scaffold was higher in the experimental group than in the control group. The time of icing effect of type I and II collagen solution was 136.54 ± 5.26 and 144.40 ± 6.45 s, respectively. The experimental scaffold had a more regular structure with actively proliferating chondrocytes that possessed adherent pseudopodia. The findings indicated that the vacuum negative pressure method did not affect the physical or chemical properties of collagen, and these scaffolds exhibited good biocompatibility with chondrocytes.
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Colágeno , Alicerces Teciduais , Alicerces Teciduais/química , Sucção , Colágeno/química , Colágeno Tipo I , Colágeno Tipo II , Engenharia Tecidual/métodosRESUMO
OBJECTIVE: Diabetic foot ulcers (DFUs) are one of the most serious diabetic consequences, leading to amputations. Various therapies have been used to treat DFUs; however, a combination of negative pressure suction, artificial skin and autogenous skin implantation have never been investigated. This study aimed to evaluate the effectiveness of a novel three-step therapy protocol using negative pressure wound therapy (NPWT), artificial skin and autogenous skin implantation in patients with DFUs. METHOD: At a single tertiary university hospital between 2015 and 2018, the three-step therapy protocol was applied to patients with DFUs and its safety and efficacy was investigated. RESULTS: A total of 21 patients took part in the study. The majority of the patients were female (62%), with a mean age of 65 years and a mean body mass index of 21kg/m2. A third (n=7) of operative sites experienced minor complications, with two requiring re-operation. At a median follow up of 24 months, the average time of complete wound healing was 46 days, and the wound healing rate was 71%. The first-stage wound healing rate was 90%. All patients had achieved remission without any further recurrence of disease. CONCLUSION: This comprehensive surgical technique for managing DFUs achieved a high local cure rate, minimal functional morbidity, and acceptable wound complication rates. The three-step therapy protocol has the potential to promote the healing process of DFUs, which is expected to serve as a new method for the treatment and cure of DFUs.
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Diabetes Mellitus , Pé Diabético , Tratamento de Ferimentos com Pressão Negativa , Pele Artificial , Idoso , Amputação Cirúrgica , Pé Diabético/terapia , Feminino , Humanos , Masculino , CicatrizaçãoRESUMO
Aim. To compare the safety and efficacy of computed tomography (CT)-assisted three-dimensional guiding templates (3DGTs) and free-hand (FH) technique for posterior cervical pedicle screw fixation in cervical spondylotic myelopathy (CSM) treatment. Methods. Thirty-five patients (216 screws) with CSM and developmental cervical stenosis were randomly divided into groups A (FH) and B (3DGTs). All patients underwent modified posterior surgery with cervical pedicle screw insertion (C1-7). Preoperative, postoperative, and intergroup comparisons of efficacy were evaluated using the visual analog scale (VAS), Japanese Orthopaedic Association (JOA), and Short Form 12 (SF-12) scores and JOA score improvement rate. Incidence of intra- and postoperative complications was analyzed. Postoperative cervical spine CT was performed to evaluate (i) the pedicle screws' deviation angle from the optimal path (sagittal deviation, α; coronal deviation angle, ß), screw insertion point's deviation distance (d), and screw accuracy and (ii) the deviation angle and distance of screw entrance point of pedicle screws from the optimal channel. Results. All patients successfully completed the procedures. Groups A and B did not significantly differ in age, sex ratio, body mass index, operative time, or intraoperative blood loss amount. Postoperative VAS, JOA, and SF-12 scores improved in both groups. VAS, JOA, or SF-12 scores did not significantly differ between the 2 groups. The α, ß, and d scores were lower in group B, but accuracy was higher in group B. Conclusions. 3DGTs and FH technique show comparable outcomes with respect to neurological improvement and safety.
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Parafusos Pediculares , Humanos , Estudos Retrospectivos , Vértebras Cervicais/diagnóstico por imagem , Vértebras Cervicais/cirurgia , Tomografia Computadorizada por Raios X/métodos , Duração da Cirurgia , Resultado do TratamentoRESUMO
Acid-sensing ion channels (ASICs) are the major proton receptor in the brain and a key mediator of acidosis-induced neuronal injuries in disease. Most of published data on ASIC function came from studies performed in mice, and relatively little is known about potential differences between human and mouse ASICs (hASIC and mASIC, respectively). This information is critical for us to better interpret the functional importance of ASICs in human disease. Here, we examined the expression of ASICs in acutely resected human cortical tissue. Compared with mouse cortex, human cortical tissue showed a similar ratio of ASIC1a:ASIC2a expression, had reduced ASIC2b level, and exhibited a higher membrane:total ratio of ASIC1a. We further investigated the mechanism for higher surface trafficking of hASIC1a in heterologous cells. A single amino acid at position 285 was critical for increased N-glycosylation and surface expression of hASIC1a. Consistent with the changes in trafficking and current, cells expressing hASIC1a or mASIC1a S285P mutant had a higher acid-activated calcium increase and exhibited worsened acidotoxicity. These data suggest that ASICs are likely to have a larger impact on acidosis-induced neuronal injuries in humans than mice, and this effect is, at least in part, a result of more efficient trafficking of hASIC1a.-Xu, Y., Jiang, Y.-Q., Li, C., He, M., Rusyniak, W. G., Annamdevula, N., Ochoa, J., Leavesley, S. J., Xu, J., Rich, T. C., Lin, M. T., Zha, X.-M. Human ASIC1a mediates stronger acid-induced responses as compared with mouse ASIC1a.
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Canais Iônicos Sensíveis a Ácido/metabolismo , Prótons , Canais Iônicos Sensíveis a Ácido/química , Canais Iônicos Sensíveis a Ácido/genética , Potenciais de Ação , Adolescente , Adulto , Animais , Células CHO , Cálcio/metabolismo , Córtex Cerebral/metabolismo , Córtex Cerebral/fisiologia , Cricetinae , Cricetulus , Feminino , Humanos , Ativação do Canal Iônico , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Pessoa de Meia-Idade , Mutação , Especificidade da EspécieRESUMO
The purpose of this paper is to analyze the properties of porcine cartilage type II collagen scaffolds crosslinked with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxy-succinamide (EDC/NHS) under different conditions. The porous EDC/NHS-crosslinked scaffolds were obtained through a two-step freeze-drying process. To determine the optimal crosslinking condition, we used different solvents and various crosslinking temperatures to prepare the scaffolds. Three crosslinking solutions were prepared with different solvents, photographs were taken with a flash in the darkroom, and light transmission was observed. Type II collagen was crosslinked on a horizontal shaker at a speed of 60 r/min according to the above grouping conditions, and then the structural change of the scaffold in each group was observed. To investigate the swelling ratio and the in vitro degradation of the collagen scaffold, tests were also carried out by immersion of the scaffolds in a PBS solution and digestion in type II collagenase, respectively. The influence of the scaffolds on the proliferation of chondrocytes was assessed by the methyl thiazolyl tetrazolium colorimetric assay. The morphology of the crosslinked scaffolds cocultured with chondrocytes was characterized by a scanning electron microscope. The results proved that 75% alcohol and a crosslinking temperature of 37 °C are recommended. Collagen fibrils are more densely packed after crosslinking with EDC/NHS and have a more uniform structure than that of noncrosslinked ones. The EDC-crosslinked scaffolds possessed excellent mechanical property and biocompatibility.
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Colágeno Tipo II/química , Reagentes de Ligações Cruzadas/química , Succinimidas/química , Alicerces Teciduais/química , Animais , Proliferação de Células , Células Cultivadas , Condrócitos/citologia , Liofilização , Coelhos , Suínos , Engenharia TecidualRESUMO
Diabetic neuropathic pain (DNP) is a prevalent complication in diabetes patients. Neuronal inflammation and activation of Toll-like receptor 4 (TLR4) are involved in the occurrence of DNP. However, the underlying mechanisms remain unclear. Downregulation of gamma-aminobutyric acid B (GABAB) receptor contributes to the DNP. GABAB receptor interacts with NF-κB, a downstream signaling factor of TLR4, in a neuropathic pain induced by chemotherapy. In this study, we determined the role of TLR4/Myd88/NF-κB signaling pathways coupled to GABAB receptors in the generation of DNP. Intrathecal injection of baclofen (GABAB receptor agonist), LPS-RS ultrapure (TLR4 antagonist), MIP (MyD88 antagonist), or SN50 (NF-κB inhibitor) significantly increased paw withdrawal threshold (PWT) and paw withdrawal thermal latency (PWTL) in DNP rats, while intrathecal injection of saclofen (GABAB receptor blocker) decreased PWT and PWTL in DNP rats. The expression of TLR4, Myd88, NF-κBp65, and their downstream components IL-1 and TNF-α was significantly higher in the spinal cord tissue in DNP rats compared to control rats. Following inhibition of TLR4, Myd88, and NF-κB, the expression of IL-1 and TNF-α decreased. Activation of GABAB receptors downregulated the expression of TLR4, Myd88, NF-κBp65, IL-1, and TNF-α. Blockade of GABAB receptors significantly upregulated expression of TLR4, Myd88, NF-κBp65, IL-1, and TNF-α. These data suggest that activation of the TLR4/Myd88/NF-κB signaling pathway is involved in the occurrence of DNP in rats. Activation of GABAB receptor in the spinal cord may suppress the TLR4/Myd88/NF-κB signaling pathway and alleviate the DNP.
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Neuropatias Diabéticas/metabolismo , Receptores de GABA-B/metabolismo , Corno Dorsal da Medula Espinal/metabolismo , Receptor 4 Toll-Like/metabolismo , Animais , Western Blotting , Ensaio de Imunoadsorção Enzimática , Imunofluorescência , Masculino , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/fisiologiaRESUMO
BACKGROUND: Large cell neuroendocrine carcinoma (LCNEC) of the bladder is a rare non-urothelial tumor of the bladder. The treatment of LCNEC of the bladder is different from that of urothelial carcinoma (UC); therefore, early and accurate diagnosis is particularly important. As LCNEC of the bladder is rare and its clinical symptoms and radiographic features are similar to those of urothelial tumors, the clinical diagnosis of the disease remains challenging. CASE SUMMARY: We report a 72-year-old female patient who presented with gross hematuria for 3 mo. A solitary tumor located in the anterior wall of the bladder was found by cystoscopy. Pathological examination after biopsy suggested UC of the bladder in the absence of immunohistochemical assessment. The patient underwent partial cystectomy and was finally diagnosed with LCNEC (pT2bN0M0) based on the results of postoperative immunohistochemical examination. During the 10-mo follow-up, no signs of tumor recurrence or metastasis were found. CONCLUSION: Immunohistochemical examination is essential for diagnosing LCNEC of the bladder. Accurate diagnosis and multidisciplinary treatment in the early stage of the disease are crucial for improving the prognosis.
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Acid-sensing ion channel-1a (ASIC1a) is a potential therapeutic target for multiple neurological diseases. We studied here ASIC1a glycosylation and trafficking, two poorly understood processes pivotal in determining the functional outcome of an ion channel. We found that most ASIC1a in the mouse brain was fully glycosylated. Inhibiting glycosylation with tunicamycin reduced ASIC1a surface trafficking, dendritic targeting, and acid-activated current density. N-glycosylation of the two glycosylation sites, Asn393 and Asn366, has differential effects on ASIC1a biogenesis. Maturation of Asn393 increased ASIC1a surface and dendritic trafficking, pH sensitivity, and current density. In contrast, glycosylation of Asn366 was dispensable for ASIC1a function and may be a rate-limiting step in ASIC1a biogenesis. In addition, we revealed that acidosis reduced the density and length of dendritic spines in a time- and ASIC1a-dependent manner. ASIC1a N366Q, which showed increased glycosylation and dendritic targeting, potentiated acidosis-induced spine loss. Conversely, ASIC1a N393Q, which had diminished dendritic targeting and inhibited ASIC1a current dominant-negatively, had the opposite effect. These data tie N-glycosylation of ASIC1a with its trafficking. More importantly, by revealing a site-specific effect of acidosis on dendritic spines, our findings suggest that these processes have an important role in regulating synaptic plasticity and determining long-term consequences in diseases that generate acidosis.
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Acidose , Espinhas Dendríticas/fisiologia , Proteínas do Tecido Nervoso/metabolismo , Neurônios/citologia , Canais de Sódio/metabolismo , Canais Iônicos Sensíveis a Ácido , Análise de Variância , Animais , Animais Recém-Nascidos , Asparagina/genética , Asparagina/metabolismo , Biotinilação/fisiologia , Células CHO , Cricetinae , Cricetulus , Feminino , Glicina/genética , Glicosilação/efeitos dos fármacos , Hipocampo/citologia , Concentração de Íons de Hidrogênio , Masculino , Potenciais da Membrana/efeitos dos fármacos , Potenciais da Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Microscopia Confocal , Mutação/genética , Proteínas do Tecido Nervoso/deficiência , Oócitos , Técnicas de Cultura de Órgãos , Técnicas de Patch-Clamp , Transporte Proteico/efeitos dos fármacos , Transporte Proteico/genética , Ratos , Canais de Sódio/deficiência , Transfecção , Tunicamicina/farmacologia , XenopusRESUMO
Background: Most previous studies on acupuncture in the treatment of knee osteoarthritis (KOA) have focused on improving functional efficacy and safety, while related mechanisms have not been systematically reviewed. Acupuncture modulates cytokines to attenuate cartilage extracellular matrix degradation and apoptosis, key to the pathogenesis of KOA, but the mechanisms are complex. Objectives: The purpose of this study is to assess the efficacy of acupuncture quantitatively and summarily in animal studies of KOA. Methods: Nine databases including PubMed, Embase, Web of Science (including Medline), Cochrane library, Scopus, CNKI, Wan Fang, and VIP were searched to retrieve animal studies on acupuncture interventions in KOA published since the inception of the journal. Relevant literature was screened, and information extracted. Meta-analysis was performed using Revman 5.4 and Stata 17.0 software. Results: The 35 included studies involved 247 animals, half of which were in acupuncture groups and half in model groups. The mean quality level was 6.7, indicating moderate quality. Meta-analysis showed that acupuncture had the following significant effects on cytokine levels in p38MAPK and mitochondrial pathways: (1) p38MAPK pathway: It significantly inhibits p38MAPK, interleukin-1beta (IL-1ß), tumor necrosis factor alpha (TNF-α), phosphorylated (p)-p38MAPK, matrix metalloproteinase-13 (MMP-13), MMP-1, a disintegrin and metalloproteinase with thrombospondin motifs-5 (ADAMST-5) expression, and significantly increased the expression of collagen II and aggrecan. (2) mitochondrial pathway: It significantly inhibited the expression of Bcl-2-associated X protein (Bax), cysteine protease-3 (caspase-3), caspase-9, and Cytochrome-c (Cyt-c). And significantly increased the expression of B cell lymphocytoma-2 (Bcl-2). In addition, acupuncture significantly reduced chondrocyte apoptosis, Mankin's score (a measure of cartilage damage), and improved cartilage morphometric characteristics. Conclusion: Acupuncture may inhibit cytokine expression in the p38MAPK pathway to attenuate cartilage extracellular matrix degradation, regulate cytokines in the mitochondrial pathway to inhibit chondrocyte apoptosis, and improve cartilage tissue-related phenotypes to delay cartilage degeneration. These findings provide possible explanations for the therapeutic mechanisms and clinical benefits of acupuncture for KOA. Systematic review registration: https://inplasy.com, identifier INPLASY20 2290125.
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Prostate cancer growth is almost wholly dependent on the route of the androgen receptor and most therapies aimed at blocking this signaling axis are useful tools in the management of this disease. Unfortunately such therapies invariably fail, and the cancer progresses to an androgen-independent stage. In such cases, androgen receptor mutation almost always occurs and much evidence suggests the continuous growth of the tumor. One mechanism by which the receptor is thought to remain active is mutation. This paper reviews the molecular mechanism of tumor growing after androgen receptor mutation and some of the promising management principles and systemic chemotherapy options against prostate cancer.
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Mutação , Neoplasias da Próstata/genética , Receptores Androgênicos/genética , Progressão da Doença , Humanos , Masculino , Neoplasias da Próstata/patologiaRESUMO
OBJECTIVE: To identify imaging markers that reflect the epidermal growth factor receptor (EGFR) mutation status by comparing computed tomography (CT) imaging-based histogram features between bone metastases with and without EGFR mutation in patients with primary lung adenocarcinoma. MATERIALS AND METHODS: This retrospective study included 57 patients, with pathologically confirmed bone metastasis of primary lung adenocarcinoma. EGFR mutation status of bone metastases was confirmed by gene detection. The CT imaging of the metastatic bone lesions which were obtained between June 2014 and December 2017 were collected and analyzed. A total of 42 CT imaging-based histogram features were automatically extracted. Feature selection was conducted using Student's t-test, Mann-Whitney U test, single-factor logistic regression analysis and Spearman correlation analysis. A receiver operating characteristic (ROC) curve was plotted to compare the effectiveness of features in distinguishing between EGFR(+) and EGFR(-) groups. DeLong's test was used to analyze the differences between the area under the curve (AUC) values. RESULTS: Three histogram features, namely range, skewness, and quantile 0.975 were significantly associated with EGFR mutation status. After combining these three features and combining range and skewness, we obtained the same AUC values, sensitivity and specificity. Meanwhile, the highest AUC value was achieved (AUC 0.783), which also had a higher sensitivity (0.708) and specificity (0.788). The differences between AUC values of the three features and their various combinations were statistically insignificant. CONCLUSION: CT imaging-based histogram features of bone metastases with and without EGFR mutation in patients with primary lung adenocarcinoma were identified, and they may contribute to diagnosis and prediction of EGFR mutation status.
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Adenocarcinoma de Pulmão/diagnóstico por imagem , Neoplasias Ósseas/diagnóstico por imagem , Neoplasias Pulmonares/diagnóstico por imagem , Mutação , Tomografia Computadorizada por Raios X/métodos , Adenocarcinoma de Pulmão/genética , Adenocarcinoma de Pulmão/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias Ósseas/genética , Neoplasias Ósseas/secundário , Receptores ErbB/genética , Feminino , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-IdadeRESUMO
It has been previously suggested that cytokeratins (CKs) are important diagnostic and prognostic biomarkers for urothelial lesions. Hence it is imperative to understand the expression pattern of cytokeratins during formation of papillary bladder cancer, which was the objective of the current study. Expression pattern of CK14 and CK18 were examined using immunohistochemical staining in a mice model of papillary bladder cancer. Twenty female mice were divided into two groups-group 1 (NT) and group 2, which received N-butyl- N-(4-hydroxybutyl) nitrosamine (BBN) for 20 weeks plus one week without treatment. Following histological classification of bladder lesions, CK14 and CK18 immunostaining was assessed according to its distribution and intensity. In NT animals, both basal cells and umbrella cells showed sporadic positive staining for CK14 and CK18, respectively. In BBN group, hyperplastic lesions showed significantly more CK14 and significantly less CK18 staining ( P < 0.05 in each case). Invasive carcinomas showed increased CK14 immunostaining in all epithelial layers. Cumulatively, our data indicate that altered CK14 (high) and CK18 (low) expression is perhaps an early event in bladder cancer tumorigenesis in females at least and is characteristic of both urothelial superficial pre-neoplastic and neoplastic lesions. Impact statement Studies have shown that expression of cytokeratins (CKs) or their altered distribution affects the bladder cancer pathogenesis and disease outcome, while the underlying mechanisms are not clear. The present study aims to explore the expression pattern of CK14 and CK18 during formation of papillary bladder cancer. The results showed that hyperplastic lesions showed significantly more CK14 and significantly less CK18 staining and invasive carcinomas showed increased CK14 immunostaining in all epithelial layers in N-butyl- N-(4-hydroxybutyl)nitrosamine (BBN)-induced mouse model. The results indicate that altered CK14 (high) and CK18 (low) expression is perhaps an early event in bladder cancer tumorigenesis and is characteristic of both urothelial superficial pre-neoplastic and neoplastic lesions, which may provide the early diagnosis index.
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Carcinoma Papilar/patologia , Queratina-14/análise , Queratina-18/análise , Lesões Pré-Cancerosas/patologia , Neoplasias da Bexiga Urinária/patologia , Animais , Modelos Animais de Doenças , Feminino , Imuno-Histoquímica , Camundongos Endogâmicos C57BLRESUMO
Cellular metabolic reprogramming is the main characteristic of cancer cells and identification of targets using this metabolic pattern is extremely important to treat cancers, such as osteosarcoma (OS). In this study, SLIT2 and ROBO1 were upregulated in OS, and higher expression of ROBO1 was associated with worse overall survival rate. Furthermore, in vitro and in vivo experiments demonstrated that the SLIT2/ROBO1 axis promotes proliferation, inhibits apoptosis, and contributes to the Warburg effect in OS cells. Mechanistically, the SLIT2/ROBO1 axis exerted cancer-promoting effects on OS via activation of the SRC/ERK/c-MYC/PFKFB2 pathway. Taken together, the findings reveal a previously unappreciated function of SLIT2/ROBO1 signaling in OS, which is intertwined with metabolic alterations that promote cancer progression. Targeting the SLIT2/ROBO1 axis may be a potential therapeutic approach for patients with OS.
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Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Osteossarcoma/metabolismo , Fosfofrutoquinase-2/metabolismo , Proteínas Proto-Oncogênicas c-myc/metabolismo , Receptores Imunológicos/metabolismo , Quinases da Família src/metabolismo , Animais , Glicólise , Xenoenxertos , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/genética , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Proteínas do Tecido Nervoso/genética , Osteossarcoma/genética , Fosforilação Oxidativa , Fosfofrutoquinase-2/genética , Proteínas Proto-Oncogênicas c-myc/genética , Receptores Imunológicos/genética , Transdução de Sinais , Quinases da Família src/genética , Proteínas RoundaboutRESUMO
Acid-sensing ion channel 1a (ASIC1a) is the key subunit that determines acid-activated currents in neurons. ASIC1a is important for neural plasticity, learning, and for multiple neurological diseases, including stroke, multiple sclerosis, and traumatic injuries. These findings underline the importance for better defining mechanisms that regulate ASIC1a expression. During the past decade, microRNA has emerged as one important group of regulatory molecules in controlling protein expression. However, little is known about whether microRNA regulates ASIC1a. Here, we assessed several microRNAs that have predicted targeting sequences in the 3' untranslated region (UTR) of mouse ASIC1a. Our results indicated that miR-144 and -149 reduced ASIC1a expression while Let-7 increased ASIC1a protein levels. miR-30c, -98, -125, -182* had no significant effect. Since a reduction in ASIC1a expression may have translational potentials in treating neuronal injury, we further asked whether the effect of miR-144 and miR-149, both reduced ASIC1a expression, was through specific targeting of the predicted sites on ASIC1a. We mutated the targeting sequence of miR-144 and miR-149 in ASIC1a UTR. The effect of miR-149 was abolished in the corresponding mutation. In contrast, miR-144 still reduced ASIC1a level when its predicted target sequence was mutated. This result indicates that miR-149 targets the 3'UTR of ASIC1a and reduces its expression.
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OBJECTIVE: To compare clinical results of treatment of Pipkin type I and II femoral head fractures through modified Smith-Peterson(S-P) approach and modified Hardinge approach. METHODS: From July 2005 to July 2014, 42 patients with Pipkin type I and II femoral head fractures were treated with operation. A total of 23 patients in anterior group was treated with modified S-P approach including 17 males and 6 females with an average age of (29.3±9.4) years old, 5 cases of type I by excision of the fragement, 3 cases of type I and 15 cases of type II cases by fixation of the fragement. While a total of 19 patients in the lateral group was treated with modified Hardinge approach including 15 males and 4 females with an average age of (31.4±10.0) years old, 3 cases of type I by excision of the fragement, 4 cases of type I and 12 cases of type II by fixation of the fragement. Operative time, blood loss during operation and fracture healing time were observed and compared. The clinical and radiographic outcomes of the patients were measured using Thompson-Epstein scoring scale. The effect of hip reduction time of less than 6 h, 6 to12 h, and more than 12 h, the effect of surgery time within 24 h and more than 24 h after injury were compared. RESULTS: All patients were followed up from 24 to 60 months with an average of(30.29±6.95) months. The operation time (61.96±12.22) min, blood loss (46.09±18.03) ml, and (74.74±10.06) min, blood loss (72.11±19.88) ml in lateral group in the anterior group were better than those of lateral group(P<0.05). In anterior group, fracture healing time was(12.22±1.70) weeks, the results were excellent in 8 cases, good in 10 cases, fair in 4 cases and poor in 1 case, the excellent and good rate was 78.3%, the incidence of avascular necrosis of femoral head was 8.69%(2/23), and the incidence of heterotopic ossification was 13.04%(3/23). While in lateral group, the fracture healing time was(12.42±1.95) weeks, the results were excellent in 6 cases, good in 7 cases, fair in 3 cases and poor in 3 cases, the excellent and good rate was 68.4%, the incidence of avascular necrosis of femoral head was 10.53%(2/19), and the incidence of heterotopic ossification was 5.26%(1/19). There was no significant difference in fracture healing time, postoperative effect and postoperative complications between the anterior group and lateral group(P<0.05). The effect of patients with reduction time of hip dislocation less than 12 h was significantly better than that of more than 12 h, there was no significant difference in the effect between reduction time within 6 h and 6 to 12 h. There was no significant difference in the outcome between surgical patients within 24 h and more than 24 h after injury. CONCLUSIONS: Dislocated hip of Pipkin type I and II femoral head fractures should be closed reduction within 6 h. If conditions are limited, the reduction time can be accepted within 12 h. Both of modified S-P approach and modified Hardinge approach are effective in treating Pipkin type I and II femoral head fractures, and can obtain excellent outcomes. Moreover, modified S-P approach has advantage of less trauma, less blood loss, shorter operative time.
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Cabeça do Fêmur/lesões , Fixação Interna de Fraturas/métodos , Luxação do Quadril/cirurgia , Fraturas do Quadril/cirurgia , Adulto , Estudos de Casos e Controles , Feminino , Fraturas do Quadril/classificação , Humanos , Masculino , Resultado do TratamentoRESUMO
Hydrogen sulfide (H2S) is an endogenous neurotransmitter that importantly regulates various physiological and pathological events including pain signal transduction. In this study, we investigated the role of spinal NMDA receptors in the nociception induced by intraplantar injection of NaHS, an H2S donor. Intraplantar injection of NaHS into hindpaw significantly decreased the paw withdrawal threshold (PWT) in contralateral hindpaw. However, intraplantar formalin injection did not produce PWT in contralateral hindpaw. Intrathecal injection of methemoglobin, a H2S scavenger, abolished hyperalgesia induced by NaHS. In addition, NaHS-induced hyperalgesia was partly, but significantly, attenuated by intrathecal injection of hydroxylamine, a cystathionine-ß-synthase (CBS) inhibitor. RT-PCR and western blotting analysis revealed that NR2B mRNA and protein levels were increased in the spinal dorsal horn, but not in dorsal root ganglion (DRG) in rats subjected to NaHS intraplantar injection. Collectively, these data suggest that peripheral injection of H2S donor causes hyperalgesia through increase in NR2B expression and production of H2S in the spinal cord.
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Sulfeto de Hidrogênio/toxicidade , Hiperalgesia/induzido quimicamente , Hiperalgesia/metabolismo , Receptores de N-Metil-D-Aspartato/biossíntese , Medula Espinal/metabolismo , Regulação para Cima/fisiologia , Animais , Sulfeto de Hidrogênio/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Medula Espinal/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacosRESUMO
BACKGROUND: Acid-sensing ion channels (ASICs) are key mediators of acidosis-induced responses in neurons. However, little is known about the relative abundance of different ASIC subunits in the brain. Such data are fundamental for interpreting the relative contribution of ASIC1a homomers and 1a/2 heteromers to acid signaling, and essential for designing therapeutic interventions to target these channels. We used a simple biochemical approach and semi-quantitatively determined the molar ratio of ASIC1a and 2 subunits in mouse brain. Further, we investigated differential surface trafficking of ASIC1a, ASIC2a, and ASIC2b. RESULTS AND CONCLUSIONS: ASIC1a subunits outnumber the sum of ASIC2a and ASIC2b. There is a region-specific variation in ASIC2a and 2b expression, with cerebellum and striatum expressing predominantly 2b and 2a, respectively. Further, we performed surface biotinylation and found that surface ASIC1a and ASIC2a ratio correlates with their total expression. In contrast, ASIC2b exhibits little surface presence in the brain. This result is consistent with increased co-localization of ASIC2b with an ER marker in 3T3 cells. Our data are the first semi-quantitative determination of relative subunit ratio of various ASICs in the brain. The differential surface trafficking of ASICs suggests that the main functional ASICs in the brain are ASIC1a homomers and 1a/2a heteromers. This finding provides important insights into the relative contribution of various ASIC complexes to acid signaling in neurons.
Assuntos
Canais Iônicos Sensíveis a Ácido/metabolismo , Encéfalo/metabolismo , Subunidades Proteicas/metabolismo , Células 3T3 , Animais , Membrana Celular/metabolismo , Retículo Endoplasmático/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Multimerização Proteica , Transporte Proteico , Reprodutibilidade dos TestesRESUMO
OBJECTIVE: To investigate the relationship of upper cervical pedicle and vertebral artery (VA) location in order to improve the safety of transpedicular screw insertion. METHODS: The vertebral arteries on 12 sides of 6 adult pate cadaverous specimens were dissected. The distance between VA and VA groove at the atlas needling point of transpedicle screw, and the distance between VA and the inner edge of axis cervical foramen, and the VA external diameter in axis cervical foramen were measured respectively. RESULTS: The distance between VA and VA groove was (1.96 ± 0.72) mm on the left and (1.99 ± 0.61)mm on the right at the atlas needling point of transpedicle screw, the distance between VA and the inner edge of axis cervical foramen was (2.23 ± 0.43) mm on the left and (2.30 ± 0.39) mm on the right, the VA external diameter in axis cervical foramen was (3.03 ± 0.48) mm on the left and (2.98 ± 0.75) mm on the right. CONCLUSION: It is unlikely to injury VA when the transpedicle screws of upper cervical vertebrae were implanted correctly besides high straddled VA, and the individualization must be performed in the process.
Assuntos
Vértebras Cervicais/cirurgia , Fixação Interna de Fraturas/métodos , Parafusos Pediculares , Artéria Vertebral/anatomia & histologia , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
OBJECTIVE: To analyze the clinical performance of TP antibody detection by CLIA kits and evaluate whether the CLIA kits made in China is suitable for clinical use. METHODS: 1200 samples were collected from Beijing Hospital including 300 samples with confirmed TP infection and 900 healthy control samples. To detect the TP antibody of the 1200 sanples separately by the CLIA kits and the ELISA kits at the same time. The test results were analyzed with statistical methods. RESULTS: The sensitivity and specificity of the CLIA kits were 99.3% and 99.9% respectively, and positive predictive value of 99.7%, negative predictive value of 100%. With the ELISA method, the positive coincidence rate was 98.7%, the negative coincidence rate was 99.8%, and the total coincidence rate was 99.5%. CONCLUSION: The CLIA kits showed good clinical performance and the agreement rate with the ELISA kits was. The CLIA kits are suitable for clinical use.