RESUMEN
The nonaqueous catalysis of lipases is significant for synthesis of high pure esters, but they usually behave low catalytic activity due to denaturation and aggregation of enzyme protein in organic phases. To improve the nonaqueous catalysis, the inexpensive copper phthalocyanine was taken as a new carrier on which Pseudomonas cepacia lipase was immobilized by physical absorption, and used for synthesis of hexyl acetate, an important flavor, via transesterification of hexanol and vinyl acetate. Results showed that the desired loading was 10-mg lipase immobilized on 10-mg copper phthalocyanine powder. When the immobilized lipase was employed in the reaction system consisted of 1.5-mL hexanol and 1.5-mL vinyl acetate at 37°C and 160 rpm, the conversion was fivefolds of that catalyzed by native lipase after 1 h, and reached 99.0% after 8 h. In six times of 8-h reuses, the immobilized lipase behaved an activity attenuation rate 1.22% h-1, lower than 1.77% h-1 of native lipase, which meant that the immobilized lipase was more stable. Even at the room temperature and the static state without shaking or stirring, the immobilized lipase still brought conversion 42.8% after 10 h and the native lipase gave 20.1%. Obviously, the immobilized lipase is an available biocatalyst in organic phase and has great potential in food industry.
RESUMEN
The nonaqueous catalysis of lipases is significant for synthesis of high pure esters, but they usually behave low catalytic activity due to denaturation and aggregation of enzyme protein in organic phases. To improve the nonaqueous catalysis, the inexpensive copper phthalocyanine was taken as a new carrier on which Pseudomonas cepacia lipase was immobilized by physical absorption, and used for synthesis of hexyl acetate, an important flavor, via transesterification of hexanol and vinyl acetate. Results showed that the desired loading was 10 mg lipase immobilized on 10 mg copper phthalocyanine powder. When the immobilized lipase was employed in the reaction system consisted of 1.5 mL hexanol and 1.5 mL vinyl acetate at 37â and 160 rpm, the conversion was five fold of that catalyzed by native lipase after 1 h, and reached 99.0% after 8 h. Undergoing six times of 8-h reuses, the immobilized lipase had an activity attenuation rate 1.22% h- 1, lower than 1.77% h- 1 of native lipase, which meant that the immobilized lipase was more stable. Even at the room temperature and the static state without shaking or stirring, the immobilized lipase could bring conversion 42.8% after 10 h and the native lipase gave 20.1%. Obviously, the immobilized lipase is an available biocatalyst in organic phase and has great potential in food industry.
Asunto(s)
Burkholderia cepacia , Lipasa/metabolismo , Enzimas Inmovilizadas/metabolismo , Ésteres , Esterificación , Catálisis , HexanolesRESUMEN
Urothelial cancer is the most frequently diagnosed type of malignant tumor in the bladder, of which primary adenocarcinoma accounts for a small percentage. Secondary malignancies, in particular metastatic adenocarcinoma from the lung, are exceedingly rare, with only six cases previously reported in the literature. The present study describes the case of a 71-year-old Chinese male patient with known lung cancer for >2 years, who was diagnosed with metastatic adenocarcinoma to the bladder. The histopathological characteristics and immunohistochemical features of the patient are reported. It was proposed that pathologists should consider the possibility of metastatic adenocarcinoma from the lung, rather than assume a diagnosis of primary adenocarcinoma of the bladder or direct invasion of adenocarcinoma from the surrounding organs. Furthermore, it is essential to determine the medical history of each patient and observe the immunohistochemical features of all tumors prior to diagnosis.
RESUMEN
Cytoskeletal reorganization is essential to keratinocyte function. Rac1 regulates cytoskeletal reorganization through signaling pathways such as the cofilin cascade. Cofilin severs actin filaments after activation by dephosphorylation. Rac1 was knocked out in mouse keratinocytes and it was found that actin filaments disassembled. In the epidermis of mice in which Rac1 was knocked out only in keratinocytes, cofilin phosphorylation was aberrantly elevated, corresponding to repression of the phosphatase slingshot1 (SSH1). These effects were independent of the signaling pathways for p21-activated kinase/LIM kinase (Pak/LIMK), protein kinase C, or protein kinase D or generation of reactive oxygen species. Similarly, when actin polymerization was specifically inhibited or Rac1 was knocked down, cofilin phosphorylation was enhanced and SSH1 was repressed. Repression of SSH1 partially blocked actin depolymerization induced by Rac1 depletion. Therefore, aberrant cofilin phosphorylation that induces actin polymerization might be a consequence of actin disassembly induced by the absence of Rac1.
Asunto(s)
Factores Despolimerizantes de la Actina/metabolismo , Actinas/metabolismo , Proteína de Unión al GTP rac1/metabolismo , Factores Despolimerizantes de la Actina/genética , Animales , Células Cultivadas , Queratinocitos/metabolismo , Ratones , Fosforilación , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/genética , Transducción de Señal/fisiología , Proteína de Unión al GTP rac1/genéticaRESUMEN
Liposarcoma has previously been described in Western studies, however, such cases are rarely reported in the mediastinum. In addition, the presence of a liposarcoma with smooth muscle and neural differentiation has not been previously reported. Thus, the present study describes the rare case of a 28-year-old Chinese male admitted to our hospital with the symptoms of chest tightness and shortness of breath due to a recurrent fibrolipoma in the mediastinum. The resected tumor measured 23 cm at its largest diameter, with histopathological and immunohistochemical features indicating a well-differentiated liposarcoma accompanied by smooth muscle and neural differentiation. Following the resection, the patient underwent radiation treatment and remains alive with no evidence of disease recurrence at two months post-surgery. To the best of our knowledge, the present study is the first to report a case of liposarcoma with smooth muscle and neural differentiation, which indicates that liposarcomas could potentially originate from stem cells. The present study highlights the fact that pathologists must carefully investigate the histopathological characteristics of liposarcomas in order to obtain an accurate diagnosis.
RESUMEN
Paraesophageal bronchogenic cyst was one of common mediastinal congenital cystic lesions of foregut origin. Because of an intimate embryologic relationship with the esophagus, they were usually found intramural (intramural esophageal bronchogenic cysts) with the local esophageal mucosa being intact and the paraesophageal bronchogenic cysts were rarely communicated with esophageal lumen. We report a case of para-esophageal bronchogenic cyst communicating to the esophageal lumen thorough a pedicle of canal, which looked liked a diverticulum on X-ray. During the operation, a communication of paraesophageal bronchogenic cyst with esophageal was found and the pathology diagnosis were made then. The symptoms of chest pain and dysphagia were relieved immediately after operation. The follow-up was well 2 years after the surgery.
Asunto(s)
Quiste Broncogénico/patología , Quiste Esofágico/patología , Quiste Broncogénico/diagnóstico , Quiste Broncogénico/cirugía , Diagnóstico Diferencial , Divertículo Esofágico/diagnóstico , Divertículo Esofágico/patología , Quiste Esofágico/diagnóstico , Quiste Esofágico/cirugía , Esófago/diagnóstico por imagen , Esófago/patología , Esófago/cirugía , Femenino , Humanos , Persona de Mediana Edad , RadiografíaRESUMEN
OBJECTIVE: To evaluate dendritic cells induced immune response against hepatocellular carcinoma by pulsed CD34+ hematopoietic stem cells originated dendritic cells with p53 gene. METHODS: CD34+ hematopoietic stem cells were harvested after mobilization by chemotherapy and G-CSF. CD34+ hematopoietic stem cell apheresis was induced to differentiate into dendritic cells by cytokine cocktail IL-4,GM-CSF and TNF-alpha. On day 7, dendritic cells were transfected with plasmid pEGFP-C3/p53 DNA. The CTL response triggered by p53 pulsed dendritic cells was assayed by MTT method. RESULTS: Dendritic cells originated from CD34+ cell apheresis had typical dendritic stick and expressed high level CD1a, CD11c, CD80, CD86, and HLA-DR molecules. After being pulsed with p53 gene, dendritic cells expressed green fluorescence protein and immunofluorescence assay (Cy3 labeled anti-P53 antibody) showed that transfected dendritic cells emitted red fluorescence. Dendritic cells inducing CTL response against HMCC97 cells (P53 positive) and HepG2 cells (P53 negative) were assessed by MTT method. P53 pulsed dendritic cells could induce P53 specific immune response against HMCC97 cells and the cytotoxin rate was (49.3+/-4.6)% compared with pEGFP-C3 transfection group [(25.4+/-4.1)%] and control group [(24.8+/-3.8)%] (P < 0.05). However, P53 pulsed dendritic cells could not induce specific CTL against P53 expression negative HepG2 cells, which the cytotoxin rates were (30.8+/-4.6)%, (27.3+/-4.3)%, and (28.5+/-5.1)% respectively in pEGFP-C3/P53 transfection group, pEGFP-C3 transfection group and control group (P > 0.05). CONCLUSION: P53 gene transfecting hematopoietic stem cell apheresis originated dendritic cells could induce specific CTL response against P53-expressing hepatocellular carcinoma cells. P53 may be a potential candidate for dendritic cell based immunotherapy of cancer.
Asunto(s)
Células Dendríticas/inmunología , Células Madre Hematopoyéticas/citología , Neoplasias Hepáticas/patología , Proteína p53 Supresora de Tumor/genética , Carcinoma Hepatocelular/inmunología , Carcinoma Hepatocelular/patología , Diferenciación Celular , Línea Celular Tumoral , Células Dendríticas/citología , Humanos , Inmunoterapia , Neoplasias Hepáticas/inmunología , Linfocitos T Citotóxicos/inmunología , TransfecciónRESUMEN
Dendritic cell (DC)-based immunotherapy is rapidly emerging as a promising treatment in cancer therapy. We had previously shown that DC pulsed with either defined mRNA of tumor antigen (Ag) such as alpha-fetoprotein (AFP), or total RNA of hepatocellular carcinoma (HCC) could elicit Ag-specific cytotoxic T lymphocyte (CTL) response. Therefore, we suggested a novel DC-based therapeutic method, in which DCs derived from CD34(+) cells enriched peripheral blood mononuclear cells were pulsed with liposome-coated AFP and mutant P53 (mtP53) fused gene pEGFP-C3/AFP-mtP53 to induce bi-targeted specific CTL responses against HCC. Three different genotype HCC cell lines, HepG2 (human histocompatibility leukocyte antigens (HLA) A2 positive, AFP expressing positive, P53 expressing negative), SMMC7721 (HLA A2 positive, neither AFP nor P53 expressing positive), and HMCC97 (HLA A2 positive, both AFP and P53 expressing positive) were selected as targets for CTL responses. An important finding was that DCs pulsed with the liposome-coated fused gene could evoke more intensive bi-targeted Ag-specific CTL responses against HMCC97 than DCs pulsed with either AFP or P53 single gene (P < 0.05). This experimental therapeutic model provides a new promising cytotherapeutic approach, in that DCs pulsed with the fused gene of different Ags might induce more extensive multitargeted antitumor immunity.
Asunto(s)
Fusión Artificial Génica , Células Dendríticas/inmunología , Genes p53 , Neoplasias Hepáticas/terapia , Mutación , Linfocitos T Citotóxicos/inmunología , alfa-Fetoproteínas/genética , Línea Celular Tumoral , Movilización de Célula Madre Hematopoyética , Humanos , Inmunoterapia , Liposomas , Neoplasias Hepáticas/inmunología , alfa-Fetoproteínas/análisisRESUMEN
BACKGROUND: DC vaccination with the use of tumor cells provides the potential to generate a polyclonal immune response to multiple known and unknown tumor Ag. Our study comparatively analyzed DC fused with tumor cells or transfected with tumor total RNA as potential cancer vaccines against hepatocellular carcinoma (HCC). METHODS: Immature DC generated from PBMC of patients with HCC were fused with HepG2-GFP (HepG2 cell line transfected stably with plasmid pEGFP-C3) cells or transfected with their total RNA. Matured DC were used to stimulate autologous T cells, and the resultant Ag-specific effector T cells were analyzed by IFN-gamma ELISPOT assay. RESULTS: DC were capable of further differentiation into mature DC after fusion with HepG2-GFP cells or transfection with HepG2-GFP cell total RNA, and were able to elicit specific T-cell responses in vitro. Both methods of Ag loading could result in stimulating CD4+ and CD8+ T cells, but with the indication that fusion loading was more efficient than RNA loading in priming the Th1 response, while RNA loading was more effective in CTL priming. DISCUSSION: Our results indicate that DC fused with tumor cells or transfected with tumor total RNA represent promising strategies for the development of cancer vaccines for treatment of HCC. They may have potential as an adjuvant immunotherapy for patients with HCC.
Asunto(s)
Vacunas contra el Cáncer/uso terapéutico , Carcinoma Hepatocelular/terapia , Células Dendríticas/química , Células Dendríticas/inmunología , Neoplasias Hepáticas/terapia , Neoplasias/metabolismo , ARN Neoplásico/genética , Carcinoma Hepatocelular/inmunología , Diferenciación Celular , Fusión Celular , Línea Celular Tumoral , Antígeno HLA-A2/inmunología , Humanos , Interleucina-1/metabolismo , Interleucina-12/metabolismo , Fenotipo , Linfocitos T Citotóxicos/inmunología , TransfecciónRESUMEN
alpha-Fetoprotein (AFP) may be a possible target for a hepatocellular carcinoma (HCC)-specific vaccination. But some studies have demonstrated that dendritic cells (DCs) treated with AFP become dysfunctional. So in this study, we try to transfect AFP mRNA into DCs and observe the ability of DCs to induce AFP-specific CD4(+) and CD8(+) T cells. We hope that AFP can be processed and presented by DCs directly, rather than released to the cultures. So there will be no AFP negative effect on the function of DCs. In the study, immature DCs generated from peripheral blood mononuclear cells (PBMCs) of HLA-A2(+) HCC patients were transfected with AFP mRNA. Then the transfected, matured DCs were used to stimulate autologous T cells. The results showed that the expressions of membrane molecules of DCs after transfection were increased dramatically, and interleukin-12 (IL-12) p70 release in the supernatant was elevated significantly. There was only a minority of AFP release in the supernatants of transfected DCs. CTLs induced by the transfected DCs recognized HLA-matched AFP positive HepG2 cell line specifically and the AFP-specific proliferative T-cell responses could also be induced. These findings indicate that this AFP mRNA transfection strategy could generate fully functional DCs, which could induce specific T cells to recognize AFP(+) HCC cells.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Células Dendríticas/inmunología , Epítopos de Linfocito T/inmunología , Activación de Linfocitos/inmunología , ARN , Transfección , alfa-Fetoproteínas/inmunología , Presentación de Antígeno/genética , Presentación de Antígeno/inmunología , Carcinoma Hepatocelular/inmunología , Carcinoma Hepatocelular/terapia , Línea Celular Tumoral , Pruebas Inmunológicas de Citotoxicidad , Células Dendríticas/metabolismo , Humanos , Neoplasias Hepáticas/inmunología , Neoplasias Hepáticas/terapia , ARN/genética , alfa-Fetoproteínas/genéticaRESUMEN
Dendritic cell (DC) vaccination with the use of total tumor RNA provides the potential to generate a polyclonal immune response to multiple known and unknown tumor antigens without HLA restriction. Our study evaluated this approach as potential immunotherapy for patients with hepatocellular carcinoma (HCC). Immature DCs generated from peripheral blood mononuclear cells of patients with HCC were transfected with HepG2-GFP (HepG2 cells transfected stably with plasmid pEGFP-C3) cells total RNA. Transfected, matured DCs were used to stimulate autologous T cells. Results revealed that DCs transfected with HepG2-GFP cells total RNA expressed EGFP when observed by flow cytometry. Compared with those before transfection, the expressions of membrane molecules were increased dramatically, and interleukin-12p70 release in the supernatant was elevated significantly. Specific T cells generated by DCs transfected with HepG2-GFP total RNA recognized HLA-matched HepG2 cell lines specifically. These findings indicate that these RNA-transfected DCs successfully generate specific T cells that specifically recognize HCC cells. Total tumor RNA-pulsed DCs may have potential as an adjuvant immunotherapy for patients with HCC.
Asunto(s)
Carcinoma Hepatocelular/inmunología , Células Dendríticas/inmunología , Inmunoterapia Adoptiva/métodos , Neoplasias Hepáticas/inmunología , Linfocitos T Citotóxicos/inmunología , Antígenos CD/inmunología , Carcinoma Hepatocelular/terapia , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Antígeno HLA-A2/inmunología , Humanos , Inmunofenotipificación , Interferón gamma/inmunología , Interleucina-12/inmunología , Células K562 , Neoplasias Hepáticas/terapia , ARN Neoplásico/genética , Subgrupos de Linfocitos T/inmunología , TransfecciónRESUMEN
AIM: To construct a recombined human AFP eukaryotic expression vector for the purpose of gene therapy and target therapy of hepatocellular carcinoma (HCC). METHODS: The full length AFP-cDNA of prokaryotic vector was digested, and subcloned to the multi-clony sites of the eukaryotic vector. The constructed vector was confirmed by enzymes digestion and electrophoresis, and the product expressed was detected by electrochemiluminescence and immunofluorescence methods. RESULTS: The full length AFP-cDNA successfully cloned to the eukaryotic vector through electrophoresis, 0.9723 IU/ml AFP antigen was detected in the supernatant of AFP-CHO by electrochemiluminescence method. Compared with the control groups, the differences were significant (P<0.05). AFP antigen molecule was observed in the plasma of AFP-CHO by immunofluorescence staining. CONCLUSION: The recombined human AFP eukaryotic expression vector can express in CHO cell line. It provides experimental data for gene therapy and target therapy of hepatocellular carcinoma.