RESUMEN

The present study aimed to investigate the effects of sevoflurane post­conditioning in a rat brain cerebral ischemia­reperfusion (I/R) model and examine its possible mechanism. Rats were randomly divided into six groups: Sham control group (Sham), I/R group, sevoflurane group (Se), Toll­like receptor­4 (TLR4) inhibitor group (Tak­242), nuclear factor (NF)­κB inhibitor group (QNZ) and Sevoflurane post­conditioning combined with TLR4­NF­κB signaling pathway inhibitor group (Se + Tak­242). Morris water maze test and tetrazolium chloride staining were used to investigate the I/R injury. The nerve cell apoptosis and autophagy in cortical tissue were detected by TUNEL and transmission electron microscopy, respectively. The expression of TLR4 protein in cortical tissue was observed by immunohistochemical staining. The expression of autophagy and apoptotic associated proteins in cortical tissues and the activity of TLR4­NF­κB signaling pathway were assayed by western blot analysis. Sevoflurane post­conditioning improved the learning and memory dysfunction caused by cerebral I/R injury. The cerebral infarction area, nerve cell apoptosis and formation of autophagic vacuoles were reduced after sevoflurane administration. The expression of light chain 3II/I, Beclin­1, Bad and Cleaved­Caspase­3 proteins were inhibited and the expression of Bcl­2 protein was upregulated after sevoflurane administration. Sevoflurane post­conditioning also inhibited the TLR4 protein and NF­κB phosphorylation, and increased inhibitor of kBα phosphorylation. The treatment effect of Tak­242 and QNZ groups were not significantly different compared with the Se group (P>0.05), and the Se + Tak­242 group had the best results. The present study demonstrated that sevoflurane post­conditioning could protect middle cerebral artery occlusion­induced brain injury rats by inhibiting autophagy and apoptosis, and that its mechanism is related to the TLR4­NF­κB signaling pathway.

Asunto(s)

Apoptosis/efectos de los fármacos , Corteza Cerebral/metabolismo , Neuronas/metabolismo , Daño por Reperfusión/tratamiento farmacológico , Sevoflurano/farmacología , Transducción de Señal/efectos de los fármacos , Animales , Corteza Cerebral/patología , Modelos Animales de Enfermedad , Masculino , Neuronas/patología , Ratas , Ratas Sprague-Dawley , Daño por Reperfusión/metabolismo , Daño por Reperfusión/patología

RESUMEN

The present prospective, randomized, double-blinded, controlled study aimed to investigate the efficacy and safety of dexmedetomidine (DEX) combined with butorphanol for patient-controlled intravenous analgesia (PCIA) following total laparoscopic hysterectomy. A total of 88 patients undergoing total laparoscopic hysterectomy and receiving postoperative PCIA were divided into two groups following surgery. Patients received DEX 0.5 µg/kg intravenously in the DEX group or 0.9% normal saline in the control (CON) group following anesthesia induction. Postoperatively, the PCIA (10 mg butorphanol with 300 µg dexmedetomidine in the DEX group or without DEX in the CON group) was delivered as a 0.5 ml bolus (lockout interval of 15 min) with a continuous background infusion of 2 ml/h. Cardiovascular and respiratory variables, cumulative butorphanol consumption, pain scores, level of sedation, concerning adverse events and the degree of patient satisfaction were recorded for 24 h post-surgery. A total of 81 patients completed the study. Blood pressure and heart rate exhibited no significant difference between the two groups during surgery and for 24 h post-surgery. Compared with the CON group, patients in the DEX group required ~19% less butorphanol (P<0.05). During the first 24 h post-surgery, patients from the DEX group had a significantly lower visual analogue scale score at rest and movement states compared with the CON group (P<0.05). There was no significant difference in sedation score between the groups. The satisfaction scores were significantly higher in the DEX group compared with those in the CON group (P<0.05). Compared with the CON group, the DEX group exhibited a lower rate of postoperative nausea and vomiting (P<0.05). There was no occurrence of serious adverse events, including respiratory depression, hypotension, bradycardia and somnolence. In conclusion, following total laparoscopic hysterectomy, the loading dose of DEX (0.5 µg/kg) followed by a continuous infusion as an adjunct to butorphanol PCIA resulted in effective analgesia, significant butorphanol sparing and less butorphanol-induced nausea and vomiting without excessive sedation or adverse effects. The trial registration number was ChiCTR1800015675 at the Chinese Clinical Trial Registry (chictr.org.cn) and the date of registration was 4th April 2018.

RESUMEN

Superoxide dismutase (SOD) is used to manage chronic pain, including neuropathic and inflammatory pain. However, data regarding the clinical effectiveness are conflicting and the neurophysiological mechanism of SOD has yet to be elucidated. The aim of the present study was to investigate whether SOD relieved chronic central pain (CCP) following spinal cord injury (SCI) and the possible underlying mechanisms. A CCP model was established using the Allen method and the CCP of the rats was measured using the paw withdrawal threshold. SOD was administered intraperitoneally following the establishment of CCP as a result of SCI. The results demonstrated that SOD relieved CCP in rats following SCI. In addition, the expression of spinal phosphorylated N-methyl-D-aspartate(NMDA) receptor subunit 1 (pNR-1) was inhibited in the CCP rats that had been treated with SOD. These observations indicated that SOD reduced mechanical allodynia and attenuated the enhancement of spinal pNR1 expression in rats with CCP. In addition, the results indicated that superoxide, produced via xanthine oxidase, and the participation of superoxide and nitric oxide (NO) as a precursor of peroxynitrite in NMDA, were involved in the mediation of central sensitization. Therefore, the observations support the hypothesis that SOD may have a potential therapeutic role for the treatment of CCP following SCI via the manipulation of superoxide and NO.

RESUMEN

This study aims to evaluate the association of E-cadherin expression with the prognosis of oral squamous cell carcinoma (OSCC). Literature retrieval, selection and assessment, data extraction, and meta-analyses were performed according to the Revman 5.0 guidelines. In the meta-analysis, we utilized either fixed effects or random effects model to pool the HR according to the test of heterogeneity. A total of nine eligible studies included 973 OSCC patients were analyzed. Of the patients, 76.3 % had low expression of E-cadherin according to the cutoff value defined by the authors. The pooled hazard ratio (HR) of low expression of E-cadherin for overall survival (OS) was 0.65 (95 % CI 0.52 to 0.80, P<0.001); in Asian population, the HR for overall survival of the patients with reduced expression of E-cadherin was 0.84 (95 % CI 0.75 to 0.95, P=0.006), and in non-Asian population, the HR for overall survival of the patients with reduced expression of E-cadherin was 0.54 (95 % CI 0.41 to 0.69, P<0.001). Patients with reduced expression of E-cadherin appear to have a poorer OS compared with those with normal or higher expression of E-cadherin.

Asunto(s)

Cadherinas/análisis , Carcinoma de Células Escamosas/mortalidad , Neoplasias de la Boca/mortalidad , Carcinoma de Células Escamosas/química , Carcinoma de Células Escamosas/patología , Transición Epitelial-Mesenquimal , Humanos , Inmunohistoquímica , Neoplasias de la Boca/química , Neoplasias de la Boca/patología , Pronóstico , Sesgo de Publicación

RESUMEN

Melanoma antigen-encoding gene 3 (MAGE-3) is an ideal candidate for a tumor vaccine although its potency need to be increased. Heat shock proteins (HSPs) represents a potential approach for increasing the potency of DNA vaccines. In the present study, a fusion DNA vaccine composed of Mycobacterium tuberculosis HSP70 and MAGE-3 was constructed and used to immunize C57BL/6 mice against B16 or B16-MAGE-3 tumor cells. The results show that the HSP70-MAGE-3 fusion DNA vaccine enhanced the frequency of MAGE-3-specific cytotoxic T-cells as compared to the MAGE-3 DNA vaccine or the HSP70/MAGE-3 cocktail DNA vaccine (P < 0.05). In conclusion, the results indicate that the HSP70-MAGE-3 fusion DNA vaccine can strongly activate MAGE-3 specific cellular immunological reactions and thus significantly inhibit the growth of B16-MAGE-3 tumors, improving the survival of tumor-bearing mice, and the HSP70-MAGE-3 fusion DNA vaccine has a significant therapeutic effect on the tumors that express MAGE-3 antigens.

Asunto(s)

Antígenos de Neoplasias/inmunología , Proteínas Bacterianas/inmunología , Vacunas contra el Cáncer/inmunología , Proteínas HSP70 de Choque Térmico/inmunología , Proteínas de Neoplasias/inmunología , Linfocitos T Citotóxicos/inmunología , Vacunas de ADN/inmunología , Animales , Anticuerpos Antineoplásicos/biosíntesis , Antígenos de Neoplasias/genética , Proteínas Bacterianas/genética , Vacunas contra el Cáncer/administración & dosificación , Vacunas contra el Cáncer/uso terapéutico , Citotoxicidad Inmunológica , Ensayos de Selección de Medicamentos Antitumorales , Proteínas HSP70 de Choque Térmico/genética , Humanos , Inmunoterapia Activa , Inyecciones Intramusculares , Interferón gamma/metabolismo , Melanoma Experimental/terapia , Ratones , Ratones Endogámicos C57BL , Proteínas de Neoplasias/genética , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , Vacunas de ADN/administración & dosificación , Vacunas de ADN/uso terapéutico

RESUMEN

AIM: To construct and express a fusion gene of human heptoma peptide (EPVTKAEML) with human heat shock protein 70 (HSP70). METHODS: A cDNA fragment encoding EPVTKAEML was added to 3 terminus of human HSP70 gene by PCR amplification. The PCR products of fusion gene were cloned into pET-28a(+)vector. The recombinant plasmid pET-28a(+)/EPVTKAEML-HSP70 was identified by enzyme digestion analysis and sequencing, and then it was transformed into E.coli BL21(DE3) through IPTG induction to express the target protein bearing His tag. RESULTS: A fragment of about 2.0 kb was amplified by PCR. Sequence analysis revealed that the sequence of EPVTKAEML was connected successfully to 3 terminus of human HSP70. Enzyme digestion analysis showed the fusion gene was cloned into pET-28a(+). SDS-PAGE showed that a relative molecular mass 72 000 fusion protein was expressed. CONCLUSION: The fusion gene of EPVTKAEML-HSP70 has been successfully constructed and expressed in E.coli BL21(DE3).

Asunto(s)

Carcinoma Hepatocelular/metabolismo , Proteínas HSP70 de Choque Térmico/metabolismo , Neoplasias Hepáticas/metabolismo , Péptidos/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , Carcinoma Hepatocelular/genética , Electroforesis en Gel de Poliacrilamida , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas HSP70 de Choque Térmico/genética , Humanos , Neoplasias Hepáticas/genética , Péptidos/genética , Reacción en Cadena de la Polimerasa , Proteínas Recombinantes de Fusión/genética

RESUMEN

MAGE-3, a member of melanoma antigen (MAGE) gene family, is recognized as an ideal candidate for tumor vaccine because it is expressed in a significant proportion of tumors of various histological types and can induce antigen-specific immune response in vivo. There is now substantial evidence that heat shock proteins HSPs isolated from cancer cells and virus-infected cells can be used as vaccines to produce cancer-specific or virus-specific immunity. In this research, we investigated whether M. tuberculosis HSP70 can be used as vehicle to elicit immune response to its accompanying MAGE-3 protein. A recombinant protein expression vector was constructed that permitted the production of fusion protein linking amino acids 195-314 of MAGE-3 to the C terminus of HSP70. We found that HSP70-MAGE-3 fusion protein can elicit stronger cellular and humoral immune responses against MAGE-3 expressing murine tumor than those elicited by MAGE-3 protein in vivo, which resulted in potent antitumor immunity against MAGE-3-expressing tumors. Covalent linkage of HSP70 to MAGE-3 was necessary to elicit immune response to MAGE-3. These results indicate that linkage of HSP70 to MAGE-3 enhanced immune responses to MAGE-3 in vivo and HSP70 can be exploited to enhance the cellular and humoral immune responses against any attached tumor-specific antigens.

Asunto(s)

Antígenos de Neoplasias/inmunología , Vacunas contra el Cáncer/inmunología , Proteínas HSP70 de Choque Térmico/inmunología , Melanoma Experimental/terapia , Mycobacterium tuberculosis/inmunología , Proteínas de Neoplasias/inmunología , Linfocitos T Citotóxicos/inmunología , Vacunas de ADN/inmunología , Animales , Células Presentadoras de Antígenos , Antígenos de Neoplasias/genética , Femenino , Proteínas HSP70 de Choque Térmico/genética , Humanos , Inmunidad Celular , Melanoma Experimental/inmunología , Ratones , Ratones Endogámicos C57BL , Proteínas de Neoplasias/genética , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , Tasa de Supervivencia , Células Tumorales Cultivadas , Vacunación

RESUMEN

AIM: To construct the melanoma antigen-1(MAGE-1) eukaryotic expression plasmid and express MAGE-1 in mouse melanoma B16 cells. METHODS: The MAGE-1 gene was amplified by PCR and cloned into the eukaryotic expression vector pIRES2-EGFP to construct the pIRES2-EGFP-MAGE-1 plasmid. The plasmid was transfected into the B16 cells. The EGFP expression was detected under fluoroscent microscope and the MAGE-1 expression was detected by immunohistochemistry staining. RESULTS: The eukaryotic expression vector pIRES2-EGFP-MAGE-1 was constructed and transfected successfully into B16 cells, and the EGFP and MAGE-1 genes were co-expressed in the B16 cells. CONCLUSION: A mouse melanoma cell line B16 co-expressing MAGE-1 and EGFP genes has been established successfully, which lays the foundation for the research on application of MAGE-1 in the tumor immunotherapy.

Asunto(s)

Línea Celular Tumoral , Proteínas Fluorescentes Verdes/biosíntesis , Melanoma Experimental/metabolismo , Proteínas de Neoplasias/biosíntesis , Animales , Antígenos de Neoplasias , Proteínas Fluorescentes Verdes/genética , Melanoma Experimental/patología , Antígenos Específicos del Melanoma , Ratones , Proteínas de Neoplasias/genética , Plásmidos , Transfección