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1.
Front Cardiovasc Med ; 10: 1126822, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37180773

RESUMO

Background: Myxomas are the most common primary cardiac tumors. Intracardiac myxomas, although benign, could cause serious consequences such as tricuspid or mitral valve obstruction, hemodynamic collapse, and acute heart failure, which pose challenges during anesthetic management. The current study was designed to summarize the anesthetic management of patients undergoing cardiac myxoma resection. Methods: This study was performed retrospectively from the perioperative period of patients who underwent myxoma resection. Patients were divided into two groups according to whether the myxoma prolapsed into the ventricle (group O) or not (group N) to evaluate the impact of tricuspid or mitral valve with obstruction. Results: 110 patients, aged 17-78 years, undergoing cardiac myxoma resection between January 2019 and December 2021 were collected, and their perioperative characteristics were recorded. In the preoperative evaluation, common clinical symptoms included dyspnea and palpitation, whereas embolic events occurred in 8 patients, including 5 (4.5%) cerebral thromboembolic events, 2 (1.8%) femoral artery, and 1 (0.9%) obstructive coronary artery. According to the echocardiography, left atrial myxoma was detected in 104 (94.5%) patients, the average dimension of myxoma was 4.03 cm ± 1.52 cm in the largest diameter, and 48 patients were divided into group O. During intraoperative anesthetic management, hemodynamic instability occurred in 38 (34.5%) patients after anesthesia induction. More patients in group O had hemodynamic instability (47.9% vs. 24.2%, p = 0.009) than in group N. The mean postoperative length of stay in the hospital was 10.64 ± 3.01 days, and most of the patients made an uneventful postoperative recovery. Conclusions: Anesthetic management for myxoma resection can be composed by assessing the myxoma, particularly the echocardiography evaluation and preventing cardiovascular instability. Typically, tricuspid or mitral valve with obstruction is a premier ingredient in anesthetic management.

2.
Neurosci Lett ; 796: 137017, 2023 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-36529387

RESUMO

BACKGROUND: Exosomes bind to and are endocytosed by neurons of various brain regions. Methods for isolating and extracting exosomes from specific brain samples are critical. At present, the most important extractive methods for exosomes are Ultracentrifugation and exosome isolation kit extraction. Both of these extraction methods have applications in neuroscience. We compare these methods to reveal the differences. METHODS: We sectioned the nucleus accumbens of mice, and isolated exosomes. A culture medium containing exosomes was extracted using ultracentrifugation (UC) and a total exosome isolation kit (TEI). The exosomes were examined using transmission electron microscopy (TEM), measurement regarding the diameter of the exosomes was done, and the thermal allodynia and western blotting analysis were also conducted, respectively. RESULTS: Transmission electron microscopy observations showed that the ultracentrifugation samples had higher purity and fewer impurities than the kit samples. The results from the two methods were then compared with a number ratio regarding the percentage was not statistically significant. Marker protein tests showed that proteins were expressed under both methods. The thermal allodynia testing observed that the two extraction methods did not affect pain behavior regarding the detection. After the kit extraction method, there were substantial white subjects suspended by PBS. CONCLUSION: Our study compared the different protocols regarding exosome extraction from the nucleus accumbens and compared the quality of two principal methods for exosome extraction from a culture medium containing exosomes. It was found that the extraction quality of exosomes by ultracentrifugation was better, but the technical difficulty was greater.


Assuntos
Exossomos , Camundongos , Animais , Exossomos/metabolismo , Núcleo Accumbens/metabolismo , Hiperalgesia/metabolismo , Proteínas/metabolismo , Meios de Cultura
3.
Acta Pharmacol Sin ; 44(5): 1095-1104, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-36418428

RESUMO

CKLF (chemokine-like factor)-MARVEL transmembrane domain containing protein 6 (CMTM6) is a novel regulator to maintain the stability of PD-L1. CMTM6 can colocalize and interact with PD-L1 on the recycling endosomes and cell membrane, preventing PD-L1 from lysosome-mediated degradation and proteasome-mediated degradation thus increasing the half-life of PD-L1 on the cell membrane. The difficulties in obtaining stable full-length PD-L1 and CMTM6 proteins hinder the research on their structures, function as well as related drug development. Using lauryl maltose neopentyl glycol (LMNG) as the optimized detergent and a cell membrane mimetic strategy, we assembled a stable membrane-bound full-length CMTM6-PD-L1 complex with amphipol A8-35. When the PD-1/PD-L1-CMTM6 interactions were analyzed, we found that CMTM6 greatly enhanced the binding and delayed the dissociation of PD-1/PD-L1, thus affecting immunosuppressive signaling and anti-apoptotic signaling. We then used the CMTM6-PD-L1 complex as immunogens to generate immune repertoires in camels, and identified a functional anti-CMTM6 nanobody, called 1A5. We demonstrated that the anti-CMTM6 nanobody greatly decreased T-cell immunosuppression and promoted apoptotic susceptibility of tumor cells in vitro, and mainly relied on the cytotoxic effect of CD8+ T-cells to exert tumor growth inhibitory effects in CT26 tumor-bearing mice. In conclusion, the stable membrane-bound full-length CMTM6-PD-L1 complex has been successfully used in studying PD-1/PD-L1-CMTM6 interactions and CMTM6-targeting drug development, suggesting CMTM6 as a novel tumor immunotherapy target.


Assuntos
Antígeno B7-H1 , Proteínas com Domínio MARVEL , Neoplasias , Anticorpos de Domínio Único , Animais , Camundongos , Antígeno B7-H1/metabolismo , Linfócitos T CD8-Positivos/metabolismo , Neoplasias/imunologia , Neoplasias/terapia , Receptor de Morte Celular Programada 1/metabolismo , Proteínas com Domínio MARVEL/imunologia , Proteínas com Domínio MARVEL/metabolismo , Engenharia de Proteínas/métodos , Anticorpos de Domínio Único/biossíntese
4.
J Clin Med ; 11(24)2022 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-36556110

RESUMO

BACKGROUND: The early detection of COVID-19 patients is fundamental for containing the pandemic. A reverse-transcriptase quantitative polymerase chain reaction (RT-PCR), which detects SARS-CoV-2 RNA, is the gold standard diagnostic test, although it can contribute to false-negative results. Consequently, supplementary diagnostic tests are urgently needed. METHODS: To assess the value of anti-SARS-CoV-2 antibody-based tests for confirming COVID-19, a retrospective study was conducted on 3120 inbound overseas travelers who underwent a 14-day government quarantine in Xiamen from August 2020 to October 2020. The diagnostic accuracy of the total antibody that detected the anti-SARS-CoV-2 antibody and the RT-PCR that detected SARS-CoV-2 RNA was determined in comparison to the clinical diagnosis. RESULTS: The COVID-19 positive rate was 3.14% (98/3120). The sensitivity and specificity of the RT-PCR test on the first day of quarantine were 14.29% and 100%, respectively, and the sensitivity and specificity of the total antibody were 93.88% and 99.40%, respectively. The kappa value between an RT-PCR on the first day of quarantine and a clinical diagnosis was 0.24 (95% CI, 0.14-0.35), indicating poor consistency. The kappa value between total antibodies and a clinical diagnosis was 0.88 (95% CI, 0.83-0.93), indicating perfect consistency. There were no differences in the positive rates of an RT-PCR in symptomatic COVID-19 (7.41% (2/27)) and asymptomatic COVID-19 (16.90 (12/71) (p = 0.338). Similarly, the positive rate of the total antibody tests showed no difference in symptomatic COVID-19 (96.30% (26/27)) and asymptomatic COVID-19 (92.96% (66/71)) (p = 0.676). CONCLUSION: SARS-CoV-2 antibodies are developed by the body in response to an infection or after vaccination; this can easily lead to a missed diagnosis. In the context of low sensitivity for an RT-PCR, SARS-CoV-2 antibody detection is an effective adjunct to RT-PCR detection, which can improve the diagnostic accuracy of COVID-19 and provide an effective complement to the false-negative results of an RT-PCR.

5.
Front Mol Neurosci ; 15: 1083671, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36590915

RESUMO

Background: Besides the established role of dopamine neurons and projections in nociceptive stimuli, the involvement of ventral tegmental area (VTA) glutamatergic projections to nucleus accumbens (NAc) in pain remains unknown. In the present study, we aimed to examine the role of VTA glutamatergic projections to NAc in painful stimuli and its related behavioral changes. Methods: Unilateral chronic constrictive injury (CCI) of sciatic nerve or intraplantar hind paw injections (i.pl.) of complete Freund's adjuvant (CFA) were used to develop pathological pain models in wild-type and VGluT2-Cre mice. The involvement of VTA glutamatergic neurons with projections to NAc in CCI-induced pain model was noted by c-Fos labeling and firing rate recordings. Pain response and pain-related behavior changes to the artificial manipulation of the VTA glutamatergic projections to NAc were observed by Hargreaves tests, von Frey tests, open field tests, elevated maze tests, and sucrose preference tests. Results: Glutamatergic neurons in VTA had efferent inputs to shell area of the NAc. The CCI pain model significantly increased neuronal activity and firing rate in VTA glutamate neurons with projections to NAc. The photoinhibition of these glutamatergic projections relieved CCI-induced neuropathic pain and CFA-induced acute and chronic inflammatory pain. Moreover, pathological neuropathic pain-induced anxiety and less sucrose preference were also relieved by inhibiting the VTA glutamatergic projections to NAc. Conclusion: Together, glutamatergic inputs from VTA to NAc contribute to chronic neuropathic and inflammatory pain and pain-related anxiety and depressive behaviors, providing a mechanism for developing novel therapeutic methods.

6.
Zhongguo Zhen Jiu ; 41(9): 993-6, 2021 Sep 12.
Artigo em Chinês | MEDLINE | ID: mdl-34491648

RESUMO

OBJECTIVE: To observe the therapeutic effect of horizontal penetration needling combined with rizatriptan monobenzoate tablets, simple horizontal penetration needling and simple rizatriptan monobenzoate tablets for migraine without aura in acute stage. METHODS: A total of 99 patients with migraine without aura in acute stage were randomized into an acupuncture plus medication group, an acupuncture group and a western medication group, 33 cases in each one. In the acupuncture group, horizontal penetration needling was applied once at Hanyan (GB 4) to Xuanli(GB 6), Shenting (GV 24) to Yintang (GV 29), Baihui (GV 20) to Qianding (GV 21), etc. for 2 h. In the western medication group, oral rizatriptan monobenzoate tablets for 10 mg were given once. In the acupuncture plus medication group, treatment of acupuncture combined with rizatriptan monobenzoate tablets were given, the application was the same as the acupuncture group and the western medication group. Before treatment and 0.5, 2, 24 h after treatment, the visual analogue scale (VAS) score was observed, the remission rate and the disappearance rate of migraine of 2, 24 h after treatment were compared in the 3 groups. RESULTS: Compared before treatment, the VAS scores of each time point after treatment were decreased in the 3 groups (P<0.05), and the changes in the acupuncture plus medication group were greater than those in the acupuncture group and the western medication group (P<0.05). The remission rates of 24 h after treatment and the disappearance rates of migraine of 2, 24 h after treatment in the acupuncture plus medication group were higher than those in the acupuncture group and the western medication group (P<0.05). CONCLUSION: Horizontal penetration needling combined with rizatriptan monobenzoate tablets have significant therapeutic effect on rapid analgesia and continuous analgesia for migraine without aura in acute stage, its effect is superior to simple horizontal penetration needling and simple rizatriptan monobenzoate tablets.


Assuntos
Terapia por Acupuntura , Enxaqueca sem Aura , Pontos de Acupuntura , Humanos , Comprimidos , Resultado do Tratamento , Triazóis , Triptaminas
7.
Acta Pharmacol Sin ; 41(3): 348-357, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31506573

RESUMO

Intestinal mucositis is a common side effect of anticancer regimens that exerts a negative impact on chemotherapy. Superoxide dismutase (SOD) is a potential therapy for mucositis but efficient product is not available because the enzyme is degraded following oral administration or induces an immune reaction after intravascular infusion. Multi-modified Stable Anti-Oxidant Enzymes® (MS-AOE®) is a new recombinant SOD with better resistance to pepsin and trypsin. We referred it as MS-SOD to distinguish from other SODs. In this study we investigated its potential to alleviate 5-FU-induced intestinal injury and the mechanisms. An intestinal mucositis model was established in C57/BL6 mice by 5-day administration of 5-FU (50 mg/kg every day, ip). MS-SOD (800 IU/10 g, ig) was given once daily for 9 days. 5-FU caused severe mucositis with intestinal morphological damage, bodyweight loss and diarrhea; MS-SOD significantly decreased the severity. 5-FU markedly increased reactive oxygen species (ROS) and inflammatory cytokines in the intestine which were ameliorated by MS-SOD. Furthermore, MS-SOD modified intestinal microbes, particularly reduced Verrucomicrobia, compared with the 5-FU group. In Caco2 cells, MS-SOD (250-1000 U/mL) dose-dependently decreased tBHP-induced ROS generation. In RAW264.7 cells, MS-SOD (500 U/mL) had no effect on LPS-induced inflammatory cytokines, but inhibited iNOS expression. These results demonstrate that MS-SOD can scavenge ROS at the initial stage of injury, thus play an indirect role in anti-inflammatory and barrier protein protection. In conclusion, MS-SOD attenuates 5-FU-induced intestinal mucositis by suppressing oxidative stress and inflammation, and influencing microbes. MS-SOD may exert beneficial effect in prevention of intestinal mucositis during chemotherapy in clinic.


Assuntos
Fluoruracila/efeitos adversos , Mucosa Intestinal/metabolismo , Superóxido Dismutase/metabolismo , Administração Oral , Animais , Fluoruracila/administração & dosagem , Fluoruracila/metabolismo , Injeções Intraperitoneais , Mucosa Intestinal/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/metabolismo , Superóxido Dismutase/administração & dosagem
8.
Cancer Manag Res ; 11: 10435-10443, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31849533

RESUMO

INTRODUCTION: CXCL14 was a significantly under-expressed mRNA in hepatocellular carcinoma tissues according to our microarray analysis, as well as head and neck squamous cell carcinoma and cervical squamous cell carcinoma. CXCL14 was considered a tumor suppressor in some studies; however, its role in HBV infection has not been identified. METHODS: CXCL14 mRNA expression was quantified from 20 male HCC patients, and the fold change in cancer tissues was calculated by comparisons with normal adjacent tissues. Overall, 212 patients with chronic HBV infection and 180 HBV-free controls were recruited to investigate the association between CXCL14 polymorphisms and HBV progression as well as liver function parameters. Serum CXCL14 levels were determined by enzyme-linked immunosorbent assay (ELISA), and comparisons were made between different HBV status and different CXCL14 genotypes. RESULTS: The mRNA expression of CXCL14 was 0.33-fold in HCC tissues when compared with adjacent tissues. The frequencies of rs2237062 and rs2547, but not rs2237061, were significantly different between patients with mild hepatitis and moderate-to-severe hepatitis. Moreover, rs2237062 and rs2547 polymorphisms correlated with impaired liver function parameters. ELISA results suggested that HBV-free controls had the highest level of CXCL14, while mild hepatitis patients had low levels, and patients with moderate-to-severe hepatitis had the lowest level. GA+AA genotypes of rs2547 were associated with reduced levels of serum CXCL14 because it introduced a stop codon at residue 109. CONCLUSION: CXCL14 was significantly suppressed in HBV-related HCC tissues, and its polymorphisms were linked with advanced stage chronic HBV infection and impaired liver function.

9.
Plant Cell Rep ; 38(6): 731-739, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30903268

RESUMO

KEY MESSAGE: OsIAAGLU could catalyze the reaction of IAA with glucose to generate IAA-glucose. Overexpression of OsIAAGLU in rice resulted in altered rice shoot architecture and root gravitropism. The distribution and levels of indole-3-acetic acid (IAA) within plant tissues are well known to play vital roles in plant growth and development. An important mechanism of regulating free IAA levels in monocots is formation of IAA ester conjugates. In this study, a cytosol-localized protein encoded by the rice gene of indole-3-acetic acid glucosyltransferase (OsIAAGLU) was found to catalyze the reaction of free IAA with glucose to generate IAA-glucose. Expression of OsIAAGLU could be induced by IAA and NAA. The number of tillers and leaf angle was significantly increased with a concomitant decrease in plant height and panicle length in the transgenic rice lines overexpressing OsIAAGLU compared to the wild-type (WT) plants. Phenotypes of iaaglu mutants constructed using the CRISPR/Cas9 system had no obvious differences with WT plants. Furthermore, overexpression of OsIAAGLU resulted in reduced sensitivity to IAA/NAA and altered gravitropic response of the roots in the transgenic plants. Free IAA contents in the leaves, root tips, and lamina joint of OsIAAGLU-overexpressing transgenic lines were lower than those of WT plants. These results support that OsIAAGLU could play a regulatory role in IAA homeostasis and rice architecture.


Assuntos
Glucose/química , Glucose/farmacologia , Ácidos Indolacéticos/química , Ácidos Indolacéticos/farmacologia , Oryza/efeitos dos fármacos , Oryza/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/genética , Oryza/genética , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/genética , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/genética , Plantas Geneticamente Modificadas/efeitos dos fármacos , Plantas Geneticamente Modificadas/genética
10.
Chin J Nat Med ; 14(11): 856-864, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27914529

RESUMO

Arisaema heterophyllum Blume is one of the three medicinal plants known as traditional Chinese medicine Rhizoma Arisaematis (RA). RA has been popularly used to treat patients with convulsions, inflammation, and cancer for a long time. However, the underlying mechanisms for RA effects are still unclear. The present study was designed to determine the cytotoxicity of agglutinin isolated from Arisema heterophyllum Blume (AHA) and explore the possible mechanisms in human non-small-cell lung cancer A549 cells. AHA with purity up to 95% was isolated and purified from Arisaema heterophyllum Blume using hydrophobic interaction chromatography. AHA dose-dependently inhibited the proliferation of A549 cells and induced G1 phase cell cycle arrest. AHA induced apoptosis by up-regulating pro-apoptotic Bax, decreasing anti-apoptotic Bcl-2, and activating caspase-9 and caspase-3. In A549 cells treated with AHA, the PI3K/Akt pathway was inhibited. Furthermore, AHA induced increase in the levels of ER stress markers such as phosphorylated eukaryotic initiation factor 2α (p-eIF2α), C/EBP-homologous protein (CHOP), inositol-requiring enzyme 1α (IRE1α), and phosphorylated c-Jun NH2-terminal kinase (p-JNK). AHA also induced autophagy in A549 cells. Staining of acidic vesicular organelles (AVOs) and increase in the levels of LC3II and ATG7 were observed in AHA-treated cells. These findings suggested that AHA might be one of the active components with anti-cancer effects in Arisaema heterophyllum Blume. In conclusion, cytotoxicity of AHA on cancer cells might be related to its effects on apoptosis and autophagy through inhibition of PI3K/Akt pathway and induction of ER stress.


Assuntos
Aglutininas/farmacologia , Apoptose/efeitos dos fármacos , Arisaema/química , Autofagia/efeitos dos fármacos , Carcinoma Pulmonar de Células não Pequenas/fisiopatologia , Medicamentos de Ervas Chinesas/farmacologia , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Células A549 , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/enzimologia , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Linhagem Celular Tumoral , Humanos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Fosfatidilinositol 3-Quinases/genética , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/genética
11.
Neuroreport ; 27(15): 1174-81, 2016 Oct 19.
Artigo em Inglês | MEDLINE | ID: mdl-27571431

RESUMO

Substantial evidence indicates that T-type Cav3.2 channel and insulin-like growth factor-1 (IGF-1) contribute to pain hypersensitivity within primary sensory nerves. A recent study suggested that activation of IGF-1 receptor (IGF-1R) could increase Cav3.2 channel currents and further contribute to inflammatory pain sensitivity. However, the expression patterns of Cav3.2 and IGF-1R and their colocalization in dorsal root ganglion (DRG) in chronic neuropathic pain condition remain unknown. In this study, we explored expression patterns of Cav3.2, IGF-1R and their colocalization, and whether phenotypic switch occurs in a subpopulation of Cav3.2 or IGF-1R neurons in mouse DRGs after sciatic nerve axotomy with immunofluorescence, real-time reverse transcription-PCR, and western blot assays. We found that expressions of Cav3.2 and IGF-1R, and their colocalization were not increased in DRGs of mice following axotomy. In addition, Cav3.2 or IGF-1R subpopulation neurons did not acquire significant switch in expression phenotype after sciatic nerve axotomy. Our findings argue for an upregulation of Cav3.2 and IGF-1R expression in lumbar DRGs post-sciatic nerve axotomy and provided an insight for understanding the functions of peripheral afferent Cav3.2 channel and IGF-1/IGF-1R signaling in chronic neuropathic pain.


Assuntos
Canais de Cálcio Tipo T/metabolismo , Gânglios Espinais/patologia , Fator de Crescimento Insulin-Like I/metabolismo , Neurônios/metabolismo , Neuropatia Ciática/patologia , Animais , Axotomia/efeitos adversos , Canais de Cálcio Tipo T/genética , Modelos Animais de Doenças , Fator de Crescimento Insulin-Like I/genética , Camundongos , RNA Mensageiro
12.
Sci Rep ; 6: 28188, 2016 06 17.
Artigo em Inglês | MEDLINE | ID: mdl-27312666

RESUMO

Increasing evidence suggests that neuro-immune and neuro-glial interactions are critically involved in chronic pain sensitization. It is well studied how immune/glial mediators sensitize pain, but how sensory neurons control neuroinflammation remains unclear. We employed Myd88 conditional knockout (CKO) mice, in which Myd88 was deleted in sodium channel subunit Nav1.8-expressing primary sensory neurons, to examine the unique role of neuronal MyD88 in regulating acute and chronic pain, and possible underlying mechanisms. We found that baseline pain and the formalin induced acute inflammatory pain were intact in CKO mice. However, the late phase inflammatory pain following complete Freund's adjuvant injection and the late phase neuropathic pain following chronic constriction injury (CCI), were reduced in CKO mice. CCI induced up-regulation of MyD88 and chemokine C-C motif ligand 2 expression in DRG neurons and macrophage infiltration into DRGs, and microglia activation in spinal dorsal horns in wild-type mice, but all these changes were compromised in CKO mice. Finally, the pain hypersensitivity induced by intraplantar IL-1ß was reduced in CKO mice. Our findings suggest that MyD88 in primary sensory neurons plays an active role in regulating IL-1ß signaling and neuroinflammation in the peripheral and the central nervous systems, and contributes to the maintenance of persistent pain.


Assuntos
Dor Crônica/patologia , Gânglios Espinais/metabolismo , Interleucina-1beta/metabolismo , Fator 88 de Diferenciação Mieloide/metabolismo , Neuralgia/patologia , Células Receptoras Sensoriais/metabolismo , Animais , Células Cultivadas , Gânglios Espinais/imunologia , Inflamação/patologia , Macrófagos/imunologia , Masculino , Camundongos , Camundongos Knockout , Microglia/metabolismo , Fator 88 de Diferenciação Mieloide/genética , Transdução de Sinais/fisiologia
13.
Neuroreport ; 27(10): 737-43, 2016 07 06.
Artigo em Inglês | MEDLINE | ID: mdl-27213932

RESUMO

Insulin-like growth factor-1 (IGF-1) is a neurotrophic factor and plays important roles in the nervous system. Increasing evidence supports that IGF-1 contributes to pain hypersensitivity through its insulin-like growth factor-1 receptor (IGF-1R) by activating IGF-1R/Akt or MAPK signaling pathways, whereas T-type Cav3.2 channel can facilitate and amplify pain signals originating from the sensory periphery. A recent study showed that activated IGF-1R can increase T-type Cav3.2 channel currents and further activate the G protein-dependent PKCα pathway to contribute to inflammatory pain sensitivity. However, the colocalization of IGF-1R and Cav3.2 in mouse dorsal root ganglion (DRG) under chronic inflammatory pain conditions remains elusive. In this study, we investigated changes in the expression of IGF-1R and the Cav3.2 channel, and their colocalization in mouse DRGs in chronic inflammatory pain condition (induced by complete Freund's adjuvant intraplanter injection) using real-time RT-PCR and immunohistochemistry approaches to confirm that Cav3.2 channel can mediate pain facilitation following IGF-1/IGF-1R signaling. We found that IGF-1R was expressed extensively in DRG neurons including small-, medium-, and large-sized neurons, whereas Cav3.2 channel was expressed exclusively in small-sized DRG neurons of naive mice. Expression of Cav3.2, but not IGF-1R, and colocalization of Cav3.2 and IGF-1R were increased in lumbar (L)4-L6 primary sensory neurons in DRGs of mice in chronic inflammatory pain. Moreover, the increased colocalization of IGF-1R and Cav3.2 is exclusively localized in small- and medium-sized primary sensory neurons. Our findings provided morphological evidence that T-type Cav3.2 channel, at least partially, mediates the pain facilitation of IGF-1/IGF-1R signaling in chronic inflammatory pain condition.


Assuntos
Canais de Cálcio Tipo T/metabolismo , Gânglios Espinais/metabolismo , Fator de Crescimento Insulin-Like I/metabolismo , Dor/metabolismo , Animais , Adjuvante de Freund , Inflamação/induzido quimicamente , Masculino , Camundongos Endogâmicos C57BL , Dor/induzido quimicamente , Células Receptoras Sensoriais/metabolismo , Regulação para Cima
14.
J Virol Methods ; 228: 60-6, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26615807

RESUMO

A novel multiplex PCR assay using non-homologous oligonucleotides with locked nucleic acid (LNA) modifications as universal primers was developed and validated for the simultaneous detection of five swine viruses. The assay utilizes five virus-specific primer pairs modified at the 5' end through the addition of the universal primer sequence. In the reaction, small amounts of target templates with the 5' tail were generated and subsequently amplified through the extension of a LNA universal primer set. To validate the specificity of this assay, 27 viral target strains and 12 non-target pathogens were tested. The lower limit of detection of viral nucleic acids was 1.1-1.9 pg per reaction or 11-32 pg in a five-plex viral nucleic acid mixture. The LNA mPCR assay displayed higher analytical sensitivity and efficiency for the detection of plasmid standards compared with the conventional assay, which uses standard primers without the 5' tail. A total of 207 field samples were tested using both assays. The LNA mPCR assay provided numerically higher detection rates for all pathogens in independent samples. Moreover, the LNA mPCR assay had significantly higher detection rates in independent samples compared with the conventional assay.


Assuntos
Reação em Cadeia da Polimerase Multiplex/métodos , Oligonucleotídeos , Doenças dos Suínos/diagnóstico , Viroses/veterinária , Vírus/genética , Vírus/isolamento & purificação , Animais , Circovirus/genética , Circovirus/isolamento & purificação , Vírus da Febre Suína Clássica/genética , Vírus da Febre Suína Clássica/isolamento & purificação , Primers do DNA , Herpesvirus Suídeo 1/genética , Herpesvirus Suídeo 1/isolamento & purificação , Limite de Detecção , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Vírus da Síndrome Respiratória e Reprodutiva Suína/isolamento & purificação , Sensibilidade e Especificidade , Suínos , Doenças dos Suínos/virologia
15.
Appl Microbiol Biotechnol ; 99(2): 919-28, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25557628

RESUMO

A novel multiplex detection array based on Luminex xMAP technology was developed and validated for simultaneous detection of five major viruses causing swine reproductive diseases. By combining one-step asymmetric multiplex reverse transcription polymerase chain reaction (RT-PCR) with xMAP bead-based hybridization and flow cytometry analysis, the resulting multiplex assay was capable of detecting single and mixed infections of PRRSV, PCV-2, PRV, CSFV, and PPV in a single reaction. The assay accurately detected and differentiated 23 viral strains used in this study. The low detection limit was determined as 2.2-22 copies/µL (corresponding to 0.5-6.8 fg/µL DNA template) on plasmid constructs containing viral fragments. The intra-assay and inter-assay variances (CV%) were low that ranged from 2.5 to 5.4 % and 4.1 to 7.6 %, respectively. The assay was applied to test field samples and detected single and mixed viral infections. The detection rate was higher than that of uniplex conventional PCR and RT-PCR methods. The detection of PRRSV by the bead-based multiplex assay was comparable with a commercially available real time RT-PCR kit. The test procedure on purified DNA or RNA samples could be completed within 2 h. In conclusion, the bead-based suspension array presented here proved to be a high-throughput practical tool that provided highly specific and sensitive identification of single and multiple infections of five major viruses in pigs and boar semen.


Assuntos
Circovirus/isolamento & purificação , Vírus da Febre Suína Clássica/isolamento & purificação , DNA Viral/isolamento & purificação , Parvovirus Suíno/isolamento & purificação , Vírus da Síndrome Respiratória e Reprodutiva Suína/isolamento & purificação , RNA Viral/isolamento & purificação , Animais , Circovirus/classificação , Peste Suína Clássica/diagnóstico , Peste Suína Clássica/virologia , Primers do DNA , DNA Viral/genética , Reação em Cadeia da Polimerase Multiplex , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Manejo de Espécimes , Suínos/virologia , Doenças dos Suínos/diagnóstico , Doenças dos Suínos/virologia
16.
Plasmid ; 74: 9-14, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24864033

RESUMO

OBJECTIVES: To learn the prevalence of the primary classical broad-host-range (BHR) IncA/C, IncN, IncP, IncQ, and IncW plasmids in dominant gram-negative bacilli from inpatients in a teaching hospital in southern China. METHODS: A multiplex polymerase chain reaction based on the replicons of BHR IncA/C, IncN, IncP, IncQ, and IncW plasmids was developed and used to determine these BHR plasmids. The difference in prevalence rates among the different species from three specimens was evaluated by a binary logistic regression model and the differences between multidrug-resistant organisms (MDRO) and non-MDRO were assessed using a chi-square test. RESULTS: The average positive detection percentages of the replicons were 4.3%, 3.7%, 3.0%, 2.6%, and 1.9%, respectively, for IncN, IncP, IncQ, IncW, and IncA/C in descending order. The distribution of all five BHR plasmids did not differ significantly between specimens collected from wounds and urine, although both were significantly higher than those of sputum. The prevalence rates of all five BHR plasmids in MDROs were significantly higher than those in non-MDRO for Enterobacteriaceae; however, no significant difference was seen in non-fermenting gram-negative bacilli (NFGNB). CONCLUSIONS: BHR IncA/C, IncN, IncP, IncQ, and IncW plasmids, which occur more often in bacilli from wound and urine specimens than those of sputum, are widespread in Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Enterobacter cloacae, Pseudomonas aeruginosa, and Acinetobacter baumannii strains isolated from inpatients. The prevalence rates in MDRO are higher than those in non-MDRO for Enterobacteriaceae but not significantly different for NFGNB.


Assuntos
Bacillus/genética , Bacillus/isolamento & purificação , Especificidade de Hospedeiro/genética , Plasmídeos/genética , Acinetobacter baumannii/genética , Acinetobacter baumannii/isolamento & purificação , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Técnicas de Tipagem Bacteriana/métodos , Criança , Pré-Escolar , Farmacorresistência Bacteriana , Enterobacter cloacae/genética , Enterobacter cloacae/isolamento & purificação , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Feminino , Humanos , Lactente , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/isolamento & purificação , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Multiplex , Proteus mirabilis/genética , Proteus mirabilis/isolamento & purificação , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/isolamento & purificação , Replicon/genética , Sensibilidade e Especificidade , Manejo de Espécimes , Adulto Jovem
17.
Sci Rep ; 3: 2119, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23846153

RESUMO

The method of synthetic gauge potentials opens up a new avenue for our understanding and discovering novel quantum states of matter. We investigate the topological quantum phase transition of Fermi gases trapped in a honeycomb lattice in the presence of a synthetic non-Abelian gauge potential. We develop a systematic fermionic effective field theory to describe a topological quantum phase transition tuned by the non-Abelian gauge potential and explore its various important experimental consequences. Numerical calculations on lattice scales are performed to compare with the results achieved by the fermionic effective field theory. Several possible experimental detection methods of topological quantum phase transition are proposed. In contrast to condensed matter experiments where only gauge invariant quantities can be measured, both gauge invariant and non-gauge invariant quantities can be measured by experimentally generating various non-Abelian gauges corresponding to the same set of Wilson loops.

18.
Artigo em Chinês | MEDLINE | ID: mdl-22295540

RESUMO

OBJECTIVE: To establish a method of acutely isolating dorsal root ganglion (DRG) neurons for patch clamp study of single-channel. METHODS: DRG neurons of rats were acutely isolated by enzymatic digestion and mechanical blowing. RESULTS: The acutely isolated DRG cells were easy to form the higher sealing resistance (> 5G Omega), which lowered noise level, so that pA-level single channel currents could be recorded. CONCLUSION: The acutely isolated DRG neurons in this study are an ideal for patch-clamp study of single-channel.


Assuntos
Separação Celular/métodos , Gânglios Espinais/citologia , Ativação do Canal Iônico/fisiologia , Neurônios/citologia , Animais , Feminino , Canais Iônicos , Masculino , Neurônios/fisiologia , Técnicas de Patch-Clamp , Ratos , Ratos Sprague-Dawley
19.
Phys Rev Lett ; 102(18): 185301, 2009 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-19518883

RESUMO

We investigate the non-Abelian Josephson effect in F=2 spinor Bose-Einstein condensates with double optical traps. We propose a real physical system which contains non-Abelian Josephson effect and has very different density and spin tunneling characters compared with the Abelian case. We calculate the frequencies of the pseudo Goldstone modes in different phases between two traps, respectively, which are the crucial feature of the non-Abelian Josephson effect. We also give an experimental protocol to observe this novel effect in future experiments.

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