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HCC, also known as hepatocellular carcinoma, is a frequently occurring form of cancer with an unfavorable prognosis. This research constructed a prognostic signature related to ubiquitination and investigated its correlation with the response to immunotherapy in HCC. The Molecular Signatures Database provided a compilation of genes associated with ubiquitination. A gene signature related to ubiquitination was obtained through Cox regression using the Least Absolute Shrinkage and Selection Operator method. The genetic factors CPY26B1, MCM10, SPINK4, and TRIM54 notably impacted the outcomes of HCC. The patients were divided into two groups: one group had a high risk of poor survival while the other had a low risk but a greater chance of controlling HCC progression. Both univariate and multivariate analyses using Cox regression found the risk score to be an independent predictor of HCC prognosis. Gene set enrichment analysis (GSEA) indicated enrichment in cell cycle and cancer-related microRNAs in high-risk groups. The tumor microenvironment (TME), response to immunotherapy, and effectiveness of chemotherapy medications positively correlated with the risk score. In the high-risk group, erlotinib showed higher IC50 values compared to the low-risk group which exhibited higher IC50 values for VX-11e, AKT inhibitor VIII, AT-7519, BMS345541, Bortezomib, CP466722, FMK, and JNK-9L. The results of RT-qPCR revealed that the expression of four UEGs was higher in tumor tissue as compared to normal tissue. Based on the genes that were expressed differently and associated with ubiquitination-related tumor categorization, we have developed a pattern of four genes and a strong nomogram that can predict the prognosis of HCC, which could be useful in identifying and managing HCC.
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Carcinoma Hepatocelular , Inmunoterapia , Neoplasias Hepáticas , Ubiquitinación , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/inmunología , Neoplasias Hepáticas/terapia , Ubiquitinación/genética , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/inmunología , Carcinoma Hepatocelular/terapia , Pronóstico , Regulación Neoplásica de la Expresión Génica , Microambiente Tumoral/genética , Microambiente Tumoral/inmunología , Masculino , Femenino , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/metabolismo , TranscriptomaRESUMEN
With the popularity of Internet technology, reading has developed in the direction of digitalization and mobileization. And entering the metaverse era, both the subject and object of reading may be redefined, presenting a new developmental pattern. This process brings a crisis to reading, such as the fragmentation of reading, the obstruction of reading needs, and the replacement of classical reading. However, reading is still an important way for college students to acquire new knowledge, broaden their horizons and improve their skills. The existence of reading crises inevitably affects the academic achievement of college students. Therefore, from the perspective of university management, this paper conducts regression analysis on 1,155 effective samples of colleges and universities in Anhui Province, extracts the factors that affect college students' reading engagement, and further explores the relationship between college students' reading engagement and academic achievement. The study concluded that: (1) in terms of family reading culture, students who grow up in families with good family reading culture perform better in reading engagement. The amount of family books, family reading education and family reading atmosphere all have significant positive effects on reading time and reflective reading strategies of college students. (2) In the cultivation of reading habits in colleges and universities, the course-driven mechanism and the atmosphere stimulating mechanism have a significant positive effect on students' reading time. The course-driven mechanism, resource supporting mechanism and atmosphere stimulating mechanism have a significant positive effect on the critical reading strategy of college students. (3) In terms of reading time, it is only found that the reading time spent on paper books has a significant positive effect on college students' academic achievement and professional quality. (4) In terms of reading strategies, the replicative reading strategy only has a significant positive effect on the improvement of college students' academic achievement and professional quality. The critical reading strategy has a significant positive effect on the professional quality, general ability and career planning ability of college students.
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Pancreatic cancer is a prevalent malignancy of the digestive system and a major cause of cancer-associated deaths. Previous studies have shown that mutation in the dermokine-ß (DMKN-ß) gene causes pancreatic and colorectal cancer. The role of the carboxy-terminal domain of DMKN-ß and dermokine-α (DMKN-α) genes in cancer tumorigenesis. Herein, the role of DMKN-α in pancreatic cancer (PC) tumorigenesis and the mechanisms underlying this process were investigated. Differentially expressed genes between PC and matched normal cells were identified through RNA-seq analysis, and the corresponding protein expression levels were verified using Western blot analysis. In vivo tumor formation experiment was also performed in nude mice. We found that the DMKN-α gene was overexpressed in cancerous pancreatic cell lines compared to normal pancreatic cell lines. CCK-8, colony formation, RTCA test, wound healing, as well as transwell test showed that the overexpression of DMKN-α enhanced the proliferation, migration, invasion, and EMT of PC cells. In vivo assays confirmed that DMKN-α promotes tumorigenesis. The findings of this study show that DMKN-α is a potential oncogene for pancreatic cancer.
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Transición Epitelial-Mesenquimal , Neoplasias Pancreáticas , Animales , Carcinogénesis/genética , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Transición Epitelial-Mesenquimal/genética , Regulación Neoplásica de la Expresión Génica , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Ratones , Ratones Desnudos , Invasividad Neoplásica/genética , Neoplasias Pancreáticas/patología , Sincalida/genética , Sincalida/metabolismo , Neoplasias PancreáticasRESUMEN
Background: Tetramethylpyrazine (TMP), a potent anti-free radical and anti-inflammations substance, has been demonstrated to possess a direct vessel relaxation property. This study aimed to evaluate the effect of TMP treatment in pulmonary hypertension (PH) and test the hypothesis that TMP prevents or reverses the process of PH. Methods: Rats (n = 36) injected with 50 mg/kg of monocrotaline (MCT) subcutaneously 4 weeks to develop PH were then randomized to TMP (5 mg/kg per day) for another 4 weeks. Hemodynamics was evaluated via the right ventricle. Pulmonary vessels structural remodeling and inflammation were examined by histologic and transmission electron microscopy observation. The expression of inducible nitric oxide synthase (iNOS) and cGMP-dependent protein kinases 1 (PKG-1) was detected by immunohistochemical staining and Western blot. Generation of reactive oxygen species (ROS) and antioxidation species was measured by biochemical analyses. Results: MCT increased PH and right ventricle hypertrophy. TMP alleviated pulmonary arterial pressure elevation, leukocyte infiltration, and structural remodeling of pulmonary arterials induced by MCT successfully. TMP treatment significantly increased the PKG-1 expression and suppressed the iNOS expression. The activity of superoxide dismutase (SOD), glutathione peroxidase (GSH), and catalase (CAT) was significantly higher than control group, while malondialdehyde (MDA) levels were lower compared with MCT group. Conclusion: TMP can suppress established MCT-induced PH through the ROS/iNOS/PKG axis. The underlying mechanisms may be associated with its anti-inflammatory, antioxidant, and antiproliferative properties in pulmonary arterial.
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Hipertensión Pulmonar , Monocrotalina , Animales , Ratas , Hipertensión Pulmonar/inducido químicamente , Hipertensión Pulmonar/tratamiento farmacológico , Hipertensión Pulmonar/metabolismo , Monocrotalina/efectos adversos , Óxido Nítrico Sintasa de Tipo II/metabolismo , Proteínas Quinasas/metabolismo , Pirazinas , Ratas Sprague-Dawley , Especies Reactivas de Oxígeno/metabolismoRESUMEN
OBJECTIVE: To investigate the protective effects of high-dose ulinastatin on the vital organs in patients undergoing total arch replacement for type A aortic dissection. METHODS: Between September 2014 and March 2016, 66 patients with type A aortic dissection underwent total arch replacement at our center. Thirty-six of the patients received ulinastatin treatment at 300 000 U/8 h from admission to 3 days postoperatively and at 300 000 U/2 h during cardiopulmonary bypass surgery (UTI group), and the other 30 patients did not receive perioperative ulinastatin treatment (control group). The surgical data and blood biochemistry profiles on days 1, 3, and 5 postoperatively were compared between the two groups, and the postoperative ICU stay, re-operation for bleeding, ventilation for over 7 days, ultrafiltration for postoperative renal failure, tracheotomy, incidences of pulmonary and neurological complications and hospital death were also compared. RESULTS: s The operating time, cardiopulmonary bypass time, ACP time, cardiac arrest time, the lowest rectal temperature and frequency of bilateral and unilateral antegrade selective cerebral perfusion were similar between the two groups (P>0.05). Compared with those in the control group, patients in UTI group had lower lactate, S-100 and neuron specific enolase levels on the first postoperative day and higher OI on the 1st, 3rd, and 5th postoperative days (P<0.05), but serum creatinine, blood urea nitrogen, total bilirubin, and alanine aminotransferase levels were comparable between the two groups (P>0.05). No significant differences were found in the frequency of re-operation for bleeding, ultrafiltration for renal failure, tracheotomy, neurological complications or hospital death after the operation between the two groups, but the patients in UTI group had a shorter ICU time, a less frequent long-term ventilation and a lower incidence of pulmonary infection (P<0.05). CONCLUSION: High-dose ulinastatin offers protection on pulmonary function and lowers the specific brain injury markers in patients with type A aortic dissection after total arch replacement, but its protective effects on brain is uncertain.
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Aneurisma de la Aorta Torácica/cirugía , Disección Aórtica/cirugía , Glicoproteínas/uso terapéutico , Sustancias Protectoras/uso terapéutico , Aorta Torácica/cirugía , Temperatura Corporal , Encéfalo/efectos de los fármacos , Puente Cardiopulmonar , Circulación Cerebrovascular , Humanos , Incidencia , Ácido Láctico/sangre , Pulmón/efectos de los fármacos , Perfusión , Fosfopiruvato Hidratasa/sangre , Periodo Posoperatorio , Proteínas S100/sangre , Factores de TiempoRESUMEN
Micro-RNA (miR)-155 is a novel gene regulator with important roles in inflammation. Herein, our study aimed to explore the role of miR-155 in LPS-induced acute lung injury(ALI). ALI in mice was induced by intratracheally delivered LPS. Loss-of-function experiments performed on miR-155 knockout mice showed that miR-155 gene inactivation protected mice from LPS-induced ALI, as manifested by preserved lung permeability and reduced lung inflammation compared with wild-type controls. Bone marrow transplantation experiments identified leukocytes, but not lung parenchymal-derived miR-155-promoted acute lung inflammation. Real-time PCR analysis showed that the expression of miR-155 in lung tissue was greatly elevated in wild-type mice after LPS stimulation. In situ hybridization showed that miR-155 was mainly expressed in alveolar macrophages. In vitro experiments performed in isolated alveolar macrophages and polarized bone marrow-derived macrophages confirmed that miR-155 expression in macrophages was increased in response to LPS stimulation. Conversely, miR-155 gain-of-function in alveolar macrophages remarkably exaggerated LPS-induced acute lung injury. Molecular studies identified the inflammation repressor suppressor of cytokine signaling (SOCS-1) as the downstream target of miR-155. By binding to the 3'-UTR of the SOCS-1 mRNA, miR-155 downregulated SOCS-1 expression, thus, permitting the inflammatory response during lung injury. Finally, we generated a novel miR-155 knockout rat strain and showed that the proinflammatory role of miR-155 was conserved in rats. Our study identified miR-155 as a proinflammatory factor after LPS stimulation, and alveolar macrophages-derived miR-155 has an important role in LPS-induced ALI.
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Lesión Pulmonar Aguda/inmunología , Macrófagos/metabolismo , MicroARNs/fisiología , Lesión Pulmonar Aguda/metabolismo , Animales , Secuencia de Bases , Sitios de Unión , Células Cultivadas , Células HEK293 , Humanos , Lipopolisacáridos/farmacología , Masculino , Ratones Endogámicos C57BL , Ratones Noqueados , Interferencia de ARN , Ratas Sprague-Dawley , Análisis de Secuencia de ADNRESUMEN
Previously, we found that electromagnetic pulses (EMP) induced an increase in blood brain barrier permeability and the leakage of albumin from blood into brain tissue. Albumin is known to activate microglia cells. Thus, we hypothesised that microglia activation could occur in the brain after EMP exposure. To test this hypothesis, the morphology and secretory function of microglia cells, including the expression of OX-42 (a marker of microglia activation), and levels of TNF-α, IL-10, IL-1ß, and NO were determined in the rat cerebral cortex after EMP exposure. In addition, to examine the signalling pathway of EMP-induced microglia activation, protein and phosphorylated protein levels of p38, JNK and ERK were determined. It was found that the expression of OX-42increased significantly at 1, 6 and 12h (p<0.05) and recovered to the sham group level at 24h after EMP exposure. Levels of NO, TNF-α and IL-10 also changed significantly in vivo and in vitro after EMP exposure. The protein level of p38 and phosphorylated p38 increased significantly after EMP exposure (p<0.05) and recovered to sham levels at 12 and 24h, respectively. Protein and phosphorylated protein levels of ERK and JNK did not change. SB203580 (p38 inhibitor) partly prevented the change in NO, IL-10, IL-1ß, TNF-α levels induced by EMP exposure. Taken together, these results suggested that EMP exposure (200kV/m, 200 pulses) could activate microglia in rat brain and affect its secretory function both in vivo and in vitro, and the p38 pathway is involved in this process.
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Corteza Cerebral/citología , Campos Electromagnéticos/efectos adversos , Sistema de Señalización de MAP Quinasas , Microglía/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo , Animales , Imidazoles/farmacología , Interleucina-10/metabolismo , Interleucina-1beta/metabolismo , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Masculino , Microglía/efectos de los fármacos , Óxido Nítrico/metabolismo , Cultivo Primario de Células , Piridinas/farmacología , Ratas , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
AIM: To detect and analyze the changes on ocular surface and tear function in type II diabetic patients with proliferative diabetic retinopathy (PDR), an advanced stage of diabetic retinopathy (DR), using conventional ophthalmic tests and the high-resolution laser scanning confocal microscopy. METHODS: Fifty-eight patients with type II diabetes were selected. Based on the diagnostic criteria and stage classification of DR, the patients were divided into the non-DR (NDR) group and the PDR group. Thirty-six patients with cataract but no other ocular and systemic disease were included as non-diabetic controls. All the patients were subjected to the conventional clinical tests of corneal sensitivity, Schirmer I Test, and corneal fluorescein staining. The non-invasive tear film break-up time (NIBUT) and tear interferometry were conducted by a Tearscope Plus. The morphology of corneal epithelia and nerve fibers was examined using the high-resolution confocal microscopy. RESULTS: The NDR group exhibited significantly declined corneal sensitivity and Schirmer I test value, as compared to the non-diabetic controls (P< 0.001). The PDR group showed significantly reduced corneal sensitivity, Schirmer I test value, and NIBUT in comparison to the non-diabetic controls (P < 0.001). Corneal fluorescein staining revealed the progressively injured corneal epithelia in the PDR patients. Moreover, significant decrease in the corneal epithelial density and morphological abnormalities in the corneal epithelia and nerve fibers were also observed in the PDR patients. CONCLUSION: Ocular surface changes, including blunted corneal sensitivity, reduced tear secretion, tear film dysfunction, progressive loss of corneal epithelia and degeneration of nerve fibers, are common in type II diabetic patients, particularly in the diabetic patients with PDR. The corneal sensitivity, fluorescein staining scores, and the density of corneal epithelial cells and nerve fibers in the diabetic patients correlate with the duration of diabetes. Therefore, ocular surface of the patients with PDR should be examined regularly by conventional approaches and confocal microscopy to facilitate early diagnosis and treatment of keratopathy.
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OBJECTIVE: To evaluate the midterm follow-up results of extended release of posterior clearance in total knee arthroplasty. METHODS: A total of 120 patients with knee osteoarthritis were equally randomly assigned to the experimental group and control group, and underwent unilateral TKA from March 2010 to March 2012. In experimental group, there were 21 males and 39 females with an average age of (62.2±10.9) years old. In the control group, there were 25 males and 35 females with an average age of (64.9±11.4) years old. All the patients were performed using the anterior knee approach. During operation, after osteotomy of the tibia and the femoral condyle, extended release of the posterior knee clearance were taken in experimental group, while only the clearance of osteophyte in the posterior condyle were performed in the control group. The KSS scores including knee functional score and knee clinical score,as well as the range of motion (ROM) of patients, were compared between the two groups at midterm follow-up. RESULTS: Totally 49 patients in the experimental group and 54 patients in the control group were followed up, and the median follow-up time was 46 months. The knee functional score of patients in the experimental group was 91.3±3.4, which was better than 86.4±3.9 of patients in the control group; initiative ROM of flexion of patients in the experimental group was (133.2±5.9)°, which was better than (126.9±7.4)° of patients in the control group. There were no significant difference of knee clinical score between 86.9±4.6 of patients in the experimental group and 85.7±5.1 of patients in the control group, and the initiative ROM of extension between (0.5±1.1)° and (0.3±1.2)°. CONCLUSION: Extended release of the posterior knee clearance contributes to the knee function and initiative flexion ROM during a midterm follow-up and patients benefit.
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Artroplastia de Reemplazo de Rodilla/métodos , Osteoartritis de la Rodilla/cirugía , Anciano , Estudios de Casos y Controles , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Osteoartritis de la Rodilla/fisiopatología , Rango del Movimiento ArticularRESUMEN
Osthole, a bioactive simple coumarin derivative extracted from a number of medicinal plants, such as Cnidium monnieri and Angelica pubescens, has been shown to exert a variety of pharmacological activities and is considered to have potential therapeutic applications. In this study, we investigated the protective effects of osthole against myocardial ischemia/reperfusion (I/R) injury in rats. Male Sprague-Dawley rats were randomly assigned to 1 of 5 groups: the sham-oeprated control group (control), the vehicle group (vehicle), and 3 treatment groups, which were treated with osthole at the concentration of 1, 10 or 50 mg/kg (intraperitoneally), respectively, upon the initiation of myocardial ischemia. Treatment with osthole suppressed the formation of lipid peroxidation products, enhanced the capacities of antioxidant enzymes and inhibited the expression of inflammatory cytokines following myocardial I/R injury. Moreover, treatment with osthole reduced high-mobility group box protein 1 (HMGB1) and phosphorylated nuclear factor (NF)-κB expression in ischemic myocardial tissue. These results demonstrate the protective effects of osthole against myocardial I/R injury in rats and suggest that these effects may be associated with its antioxidant and anti-inflammatory activities.
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Bloqueadores de los Canales de Calcio/farmacología , Cumarinas/farmacología , Daño por Reperfusión Miocárdica/prevención & control , Animales , Bloqueadores de los Canales de Calcio/administración & dosificación , Catalasa/metabolismo , Cumarinas/administración & dosificación , Citocinas/metabolismo , Modelos Animales de Enfermedad , Activación Enzimática/efectos de los fármacos , Proteína HMGB1/metabolismo , Hemodinámica , Mediadores de Inflamación/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Masculino , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/patología , Daño por Reperfusión Miocárdica/fisiopatología , Miocardio/metabolismo , Miocardio/patología , FN-kappa B/metabolismo , Estrés Oxidativo/efectos de los fármacos , Ratas , Superóxido Dismutasa/metabolismoRESUMEN
Sequencing of the chloroplast (cp) genome using traditional sequencing methods has been difficult because of its size (>120 kb) and the complicated procedures required to prepare templates. To explore the feasibility of sequencing the cp genome using DNA extracted from whole cells and Solexa sequencing technology, we sequenced whole cellular DNA isolated from leaves of three Brassicarapa accessions with one lane per accession. In total, 246, 362, and 361 Mb sequence data were generated for the three accessions Chiifu-401-42, Z16, and FT, respectively. Micro-reads were assembled by reference-guided assembly using the cpDNA sequences of B. rapa, Arabidopsis thaliana, and Nicotiana tabacum. We achieved coverage of more than 99.96% of the cp genome in the three tested accessions using the B. rapa sequence as the reference. When A. thaliana or N. tabacum sequences were used as references, 99.7-99.8 or 95.5-99.7% of the B. rapa cp genome was covered, respectively. These results demonstrated that sequencing of whole cellular DNA isolated from young leaves using the Illumina Genome Analyzer is an efficient method for high-throughput sequencing of cp genome.
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DeBakey type II aortic dissection combined with true aneurysm in the distal aortic arch is a very rare condition, and due to the complicated pathology, its surgical intervention can be difficult and risky. We report a case with this pathology in a 23-year-old man, who received a previous open heart surgery for ventricular septal defect at the age of 6 years. Aortic valve, ascending aorta and total aortic arch replacements were performed. Cardiopulmonary bypass (CPB) including deep hypothermic circulatory arrest with selective antegrade cerebral perfusion was used. No postoperative complications occurred, and the follow-up examination at 6 months after the operation showed good therapeutic results.
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Aorta Torácica , Aneurisma de la Aorta Torácica/cirugía , Disección Aórtica/cirugía , Aorta Torácica/cirugía , Implantación de Prótesis Vascular/métodos , Puente Cardiopulmonar , Circulación Cerebrovascular , Paro Circulatorio Inducido por Hipotermia Profunda , Humanos , Masculino , Perfusión/métodos , Adulto JovenRESUMEN
OBJECTIVE: To evaluate the causes and therapies for the complications induced by endovenous laser treatment (EVLT) in patients with varicose vein of lower limbs. METHODS: From December 2003 to October 2009, 283 cases (totally 361 lower limbs) treated by the endovenous laser treatment or combined with other treatment were analyzed retrospectively. There were 108 male and 175 female patients, with a mean age of 52 years (ranging from 17 to 83 years). The varicose vein occurred in left limb for 112 cases, right limbs for 93 cases and both limbs for 78 cases. RESULTS: The patients were followed up for average 18 months, during which 142 limbs showed skin bruises and/or hematoma, 47 limbs showed skin burns, 7 limbs showed phlebitis; all of these cases were cured. In addition 91 limbs appeared abnormal skin sensation, 87 of them were recovered and 4 improved. Two limbs were found with deep vein thrombosis in this series, after treatment one case was obviously improved and other was alleviated. CONCLUSIONS: EVLT is a effective therapy for varicose vein of lower limbs. But it may lead to some complications without right manipulations.
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Terapia por Láser/efectos adversos , Complicaciones Posoperatorias , Várices/cirugía , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Terapia por Láser/métodos , Extremidad Inferior/irrigación sanguínea , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Resultado del Tratamiento , Adulto JovenRESUMEN
OBJECTIVE: To evaluate the therapeutic effect and safety of thoracic aortic replacement with concomitant endoluminal stent grafting in the treatment of DeBakey type I aortic dissection. METHODS: From September 2007 to January 2010, 6 patients with DeBakey type I aortic dissection (including one with aortic dissection relapse) received ascending aortic (or Bentall) and total aortic arch replacement and simultaneous stent graft implantation into the descending aorta. Multi-slice spiral CT scans (MSCT) were performed in each patient regularly after the surgery. Cardio-pulmonary bypass including deep hypothermic circulatory arrest with selective antegrade cerebral perfusion were used during the surgery. RESULTS: All the patients recovered smoothly after the surgical procedure without serious complications. The time of cardiopulmonary bypass ranged from 208 to 291 min (mean 242 min), arrest time of the ascending aortic was 112-194 min (mean 145 min), and selective cerebral perfusion time was 63-102 min (mean 76 min). The patients were followed up for 4-32 months (mean 15.5 months), and MSCT revealed smooth blood flow in the prosthesis, complete thrombus formation in the false lumen in the perigraft space and shrinkage of the distal false lumen without internal fistula or stent dislocation. CONCLUSION: Thoracic aortic replacement with concomitant endoluminal stent grafting is a safe and effective treatment of DeBakey type I dissection.
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Aneurisma de la Aorta/cirugía , Disección Aórtica/cirugía , Implantación de Prótesis Vascular/métodos , Adulto , Disección Aórtica/clasificación , Aneurisma de la Aorta/clasificación , Prótesis Vascular , Femenino , Humanos , Masculino , Persona de Mediana Edad , Stents , Resultado del TratamientoRESUMEN
The blood-testis barrier (BTB) plays an important role in male reproductive system. Lots of environmental stimulations can increase the permeability of BTB and then result in antisperm antibody (AsAb) generation, which is a key step in male immune infertility. Here we reported the results of male mice exposed to electromagnetic pulse (EMP) by measuring the expression of tight-junction-associated proteins (ZO-1 and Occludin), vimentin microfilaments, and transforming growth factor-beta (TGF-beta3) as well as AsAb level in serum. Male BALB/c mice were sham exposed or exposed to EMP at two different intensities (200kV/m and 400kV/m) for 200 pulses. The testes were collected at different time points after EMP exposure. Immunofluorescence histocytochemistry, western blotting, laser confocal microscopy and RT-PCR were used in this study. Compared with sham group, the expression of ZO-1 and TGF-beta3 significantly decreased accompanied with unevenly stained vimentin microfilaments and increased serum AsAb levels in EMP-exposed mice. These results suggest a potential BTB injury and immune infertility in male mice exposed to a certain intensity of EMP.
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Barrera Hematotesticular/metabolismo , Campos Electromagnéticos , Proteínas de la Membrana/metabolismo , Fosfoproteínas/metabolismo , Citoesqueleto de Actina/metabolismo , Animales , Anticuerpos/sangre , Anticuerpos/inmunología , Regulación de la Expresión Génica , Masculino , Ratones , Ratones Endogámicos BALB C , Ocludina , Permeabilidad , Espermatozoides/inmunología , Uniones Estrechas , Factores de Tiempo , Factor de Crecimiento Transformador beta3/metabolismo , Vimentina/metabolismo , Proteína de la Zonula Occludens-1RESUMEN
The aim of this study was to determine the role of protein kinase C signaling in electromagnetic pulse (EMP)-induced blood-brain barrier (BBB) permeability change in rats. The protein level of total PKC and two PKC isoforms (PKC-alpha, and PKC-beta II) were determined in brain cerebral cortex microvessels by Western blot after exposing rats to EMP at 200kV/m for 200 pulses with 1Hz repetition rate. It was found that the protein level of PKC and PKC-betaII (but not PKC-alpha) in cerebral cortex microvessels increased significantly at 0.5h and 1h after EMP exposure compared with sham-exposed animals and then recovered at 3h. A specific PKC antagonist (H7) almost blocked EMP-induced BBB permeability change. EMP-induced BBB tight junction protein ZO-1 translocation was also inhibited. Our data indicated that PKC signaling was involved in EMP-induced BBB permeability change and ZO-1 translocation in rat.
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Barrera Hematoencefálica/enzimología , Campos Electromagnéticos , Proteína Quinasa C/metabolismo , 1-(5-Isoquinolinesulfonil)-2-Metilpiperazina/farmacología , Animales , Barrera Hematoencefálica/efectos de los fármacos , Barrera Hematoencefálica/efectos de la radiación , Corteza Cerebral/metabolismo , Masculino , Proteínas de la Membrana/metabolismo , Microvasos/metabolismo , Permeabilidad/efectos de los fármacos , Permeabilidad/efectos de la radiación , Fosfoproteínas/metabolismo , Proteína Quinasa C/antagonistas & inhibidores , Proteína Quinasa C beta , Inhibidores de Proteínas Quinasas/farmacología , Transporte de Proteínas , Ratas , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacos , Transducción de Señal/efectos de la radiación , Proteína de la Zonula Occludens-1RESUMEN
The blood-brain barrier (BBB) is critical to maintain cerebral homeostasis. In this study, we examined the effects of exposure to electromagnetic pulse (EMP) on the functional integrity of BBB and, on the localization and expression of tight junction (TJ) proteins (occludin and ZO-1) in rats. Animals were sham or whole-body exposed to EMP at 200 kV/m for 400 pulses. The permeability of BBB in rat cerebral cortex was examined by using Evans Blue (EB) and lanthanum nitrate as vascular tracers. The localization and expression of TJ proteins were assessed by western blot and immunofluorescence analysis, respectively. The data indicated that EMP exposure caused: (i) increased permeability of BBB, and (ii) altered localization as well as decreased levels of TJ protein ZO-1. These results suggested that the alteration of ZO-1 may play an important role in the disruption of tight junctions, which may lead to dysfunction of BBB after EMP exposure.
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Barrera Hematoencefálica/efectos de la radiación , Campos Electromagnéticos , Uniones Estrechas/efectos de la radiación , Animales , Masculino , Proteínas de la Membrana/análisis , Ocludina , Permeabilidad , Fosfoproteínas/análisis , Ratas , Ratas Sprague-Dawley , Proteína de la Zonula Occludens-1RESUMEN
Pectin methylesterase (PME) is known to have important roles in pollen development and pollen tube growth. As pivotal regulatory factors in PME activity modulation, PME inhibitors (PMEIs) are thought to be key regulators of cell wall stability at the tip of the pollen tube. We report on the cloning and characterization of a novel B. oleracea PMEI gene, BoPMEI1. Heterologously expressed BoPMEI1 showed PMEI activity. RT-PCR studies of different tissues and promoter-GUS fusions confirmed that BoPMEI1 was specifically expressed in mature pollen grains and pollen tubes. Based on in vivo transient assays, we found that BoPMEI1 appears to be largely localized to the plasma membrane. Transgenic Arabidopsis plants expressing antisense BoPMEI1 under the control of the CaMV 35S promoter suppressed the expression of the orthologous gene At1g10770, which led to partial male sterility and decreased seed set by inhibition of pollen tube growth. Our study demonstrates the involvement of BoPMEI1 in pollen tube growth.
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Brassica/crecimiento & desarrollo , Hidrolasas de Éster Carboxílico/antagonistas & inhibidores , Inhibidores Enzimáticos/metabolismo , Proteínas de Plantas/metabolismo , Tubo Polínico/crecimiento & desarrollo , Tubo Polínico/metabolismo , Secuencia de Aminoácidos , Arabidopsis/citología , Arabidopsis/genética , Secuencia de Bases , Brassica/citología , Brassica/enzimología , Brassica/genética , Hidrolasas de Éster Carboxílico/metabolismo , Membrana Celular/enzimología , Electroforesis en Gel de Poliacrilamida , Regulación de la Expresión Génica de las Plantas , Germinación , Glucuronidasa/metabolismo , Resistencia a la Kanamicina/genética , Datos de Secuencia Molecular , Especificidad de Órganos , Fenotipo , Infertilidad Vegetal , Proteínas de Plantas/química , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente , Tubo Polínico/citología , Tubo Polínico/enzimología , Estructura Secundaria de Proteína , Transporte de Proteínas , ARN sin Sentido/metabolismo , Semillas/enzimología , Semillas/crecimiento & desarrolloRESUMEN
Rapid alkalinization factors (RALFs) are recently reported active peptide hormones and are considered to play important roles in plant development. We previously identified a differentially expressed cDNA fragment between cabbage flower buds of sterility lines and its maintainer line, which showed significant homology to Arabidopsis RALFL9. The novel RALF cDNA (BoRALF1) was isolated from broccoli flower buds by EST assembly. The open reading frame (ORF) comprises 240 bp, encoding a small putative preprotein of 79 amino acids (molecular weight of 8.72 kDa and a pI of 7.8), which contains the mature polypeptide at its C terminus. BoRALF1 shares 70.3% identity with Arabidopsis RALFL9, but has only moderate similarity with functionally characterized RALFs (ranging from 16.2% to 38.0%). BoRALF1 shows typical features of RALFs, including the 28-aa signal peptide, typical arrangement of four position conserved cysteines, the YIXY motif and a similar secondary structure. RT-PCR studies of different tissues and promoter-GUS fusions confirmed that BoRALF1 is expressed strictly in mature pollen grains and in the anther cells around the loculi. Based on in vivo transient assays, we found that BoRALF1 appears to be largely localized in the plasma membrane. Although the function of BoRALF1 remains to be determined, our experiments confirm the presence of RALF peptide in broccoli, and suggest it could have a role in anther or pollen development.
Asunto(s)
Brassica/genética , Regulación de la Expresión Génica de las Plantas , Genes de Plantas/genética , Proteínas de Plantas/genética , Polen/genética , Membrana Celular/metabolismo , Clonación Molecular , Glucuronidasa/metabolismo , Especificidad de Órganos/genética , Filogenia , Proteínas de Plantas/aislamiento & purificación , Proteínas de Plantas/metabolismo , Regiones Promotoras Genéticas/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
OBJECTIVE: To observe the effect of electromagnetic pulse (EMP) exposure on cerebral micro vascular permeability in rats. METHODS: The whole-body of male Sprague-Dawley rats were exposed or sham exposed to 200 pulses or 400 pulses (1 Hz) of EMP at 200 kV/m. At 0.5, 1, 3, 6, and 12 h after EMP exposure, the permeability of cerebral micro vascular was detected by transmission electron microscopy and immunohistochemistry using lanthanum nitrate and endogenous albumin as vascular tracers, respectively. RESULTS: The lanthanum nitrate tracer was limited to the micro vascular lumen with no lanthanum nitrate or albumin tracer extravasation in control rat brain. After EMP exposure, the lanthanum nitrate ions reached the tight junction, basal lamina and pericapillary tissue. Similarly, the albumin immunopositive staining was identified in pericapillary tissue. The changes in brain micro vascular permeability were transient, the leakage of micro vascular vessels appeared at 1 h, and reached its peak at 3 h, and nearly recovered at 12 h, after EMP exposure. In addition, the leakage of micro vascular was more obvious after exposure of EMP at 400 pulses than after exposure of EMP at 200 pulses. CONCLUSION: Exposure to 200 and 400 pulses (1 Hz) of EMP at 200 kV/m can increase cerebral micro vascular permeability in rats, which is recoverable.