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The Eimeria tenella Yulin strain (EtYL), which is sensitive to most anti-coccidial drugs, was isolated in the Yulin area of Guangxi, China. Then, Eimeria tenella Yulin precocious line (pEtYL), a precocious line with a prepatent period of 108 h, was obtained through early selection. The biological characteristics of pEtYL, including its morphology, purity, oocyst excretion curve, reproductive capacity, pathogenicity, immunogenicity, and preservation time, were comprehensively analyzed. The results showed that the isolated precocious line of E. tenella exhibited high purity, relatively weak pathogenicity, and good immunogenicity and can be used as a live vaccine line for chicken coccidiosis.
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Coccidiose , Eimeria tenella , Doenças das Aves Domésticas , Animais , China , Coccidiose/prevenção & controle , Oocistos , Virulência , GalinhasRESUMO
In order to find a new preservation solution for avian coccidial oocysts that can replace potassium dichromate (K2Cr2O7) solution, Eimeria tenella oocysts were preserved in 0.1 to 10% potassium sorbate (C6H7KO2) solution in this study. The results showed that there was no significant difference between the sporulation rate of E. tenella oocysts preserved in 0.1 to 10% C6H7KO2 solution and in 2.5% K2Cr2O7 solution (p > 0.05). The 0.5 to 10% C6H7KO2 solution could also effectively inhibit the growth of bacterial microorganisms. E. tenella oocysts preserved in 1% C6H7KO2 solution at 4 °C for 3, 6, 9, and 12 months, with the oocyst production of E. tenella oocysts being 1.3-, 1.2-, 1.6-, and 1.3-fold higher than that of oocysts stored in 2.5% K2Cr2O7 solution (p < 0.05). In conclusion, C6H7KO2 could replace K2Cr2O7 as the preservation solution of avian coccidial oocysts.
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Eimeria tenella/crescimento & desenvolvimento , Oocistos/crescimento & desenvolvimento , Preservação Biológica , Ácido Sórbico , Animais , Galinhas , Coccidiose , Eimeria , Doenças das Aves Domésticas/parasitologia , Esporos de ProtozoáriosRESUMO
Vibrio parahaemolyticus (V. parahaemolyticus) is a well-known food-borne human pathogen that can cause a variety of clinical manifestations after the consumption of raw or undercooked seafoods. The crucial roles of Vibrio OmpU in bacterial pathogenesis have been found in recent studies. In the present study, we screened for single domain antibody fragment (sdAb) candidates that bind to V. parahaemolyticus OmpU by using a sdAb phage display library and isolated several positive phage clones. The UAb28, which was one of the positive clones, was shown high enrichment and affinity. The CDRs of UAb28 are speculated to perform the OmpU binding function by molecular docking. The capable of recognizing OmpU was verified by binding and inhibition assays. The UAb28 might be useful in future studies to develop the potential sdAb-based immunotherapeutics against V. parahaemolyticus infection.
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Anticorpos Antibacterianos/imunologia , Antígenos de Bactérias/imunologia , Proteínas da Membrana Bacteriana Externa/imunologia , Vibrio parahaemolyticus/imunologia , Proteínas da Membrana Bacteriana Externa/genética , Ensaio de Imunoadsorção Enzimática , Simulação de Acoplamento Molecular , Biblioteca de Peptídeos , Vibrio parahaemolyticus/genéticaRESUMO
OBJECT: The purpose of this article was to synthesize the evidence regarding the reduction of physical restraint, and to seek some practical recommendations based on the current situation in China. METHOD: Nine databases were retrieved; these were PubMed, CINAHL, MEDLINE, Trip Database, PsysINFO, Cochrane Library, CNKI (Chinese database), Wanfang (Chinese database) and CBM (Chinese database) respectively. The selected articles were screened manually, and the identified researches were appraised through Review manager 5.3. RESULT: Eight studies (four randomized controlled trials and four quasi-experimental studies) published between June 2013 and May 2017 were selected. Risk ratios (RRs) with 95% confidence intervals (CIs) were used as the effect index for dichotomous variables. The standardized mean differences (SMDs) with 95% CIs were calculated as the pooled continuous effect. The outcome of meta-analysis suggested staff training reduced the duration (IV=-0.88; 95% CIs=-1.65 to -0.10; Z=2.22; p=0.03) and adverse effect (RR, 0.16; 95% CIs=0.09 to 0.30; Z=5.96; p<0.00001) of physical restraint, but there were no statistical change in the frequency of physical restraint (RR, 0.74; 95% CIs=0.43 to 1.28; Z=1.07; p=0.28). Noticeably, the result of pooled estimates from 3 RCTs suggested staff training had no effects on the incidence of physical restraint. (RR, 1.01; 95% CIs=0.45 to 2.24; Z=0.02; p=0.99) CONCLUSION: Staff training was an effective measure to minimize the duration and adverse effects of physical restraint. More studies are needed to examine the effectiveness of staff training in relation to reduce the prevalence of physical restraint. Furthermore, considering the nurse's education background in China, it is recommended to conduct a compulsory training program to reduce the unnecessary restraint.
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Prática Clínica Baseada em Evidências , Capacitação em Serviço/métodos , Serviços de Saúde Mental , Restrição Física , China , HumanosRESUMO
BACKGROUND: Pyogenic liver abscess (PLA) is a purulent disease caused by microbial contamination of liver parenchyma and includes amoebic liver abscess, fungal liver abscess, and the most common bacterial liver abscess. OBJECTIVE: Explore the efficacy of contrast-enhanced ultrasound (CEUS) via vessels and surgical drain guidance percutaneous catheter drainage (PCD) in the treatment of pyogenic liver abscesses (PLA). MATERIALS AND METHODS: A total of 86 PLA patients who underwent PCD treatment in our hospital from May 2018 to February 2023 were retrospectively selected. Of them, 41 patients were treated under intravenous CEUS guidance (Control group), and 45 patients were treated under CEUS via vessels and surgical drain guidance (study group). Perioperative characteristics, treatment effectiveness, and incidence of complications were analyzed and compared between groups. RESULTS AND DISCUSSION: The duration of surgery, drainage, white blood cell recovery, body temperature recovery, and hospitalization in the study group were longer than those in the control group (P<0.05). The total effective rate of the study group (95.56%) was higher than that of the control group (80.49%) (P<0.05). The incidence of complications in the study group (4.44%) was lower than that in the control group (19.51%) (P<0.05). CONCLUSION: Compared with intravenous CEUS alone, treatment under CEUS via vessels and surgical drains-guided PCD was associated with shorter surgical time, faster recovery, better treatment effect, lower risk of complications, and ensured treatment safety in PLA patients.
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Background: IgG4-related disease (IgG4-RD) is a systemic inflammatory disease which involves various organs such as the pancreas, lacrimal gland, salivary gland, retroperitoneum, and so on. These organs can be affected concomitantly. 18-Fluorodeoxyglucose positron emission tomography computed tomography (FDG-PETCT) is a systemic examination which can identify active inflammation and detect multiple organ involvement simultaneously. Pericardial involvement is rare in IgG4-RD, early detection and treatment can greatly improve the prognosis of patients. Case summary: We reported a 82-year-old female patient referred to our department complaining of chest tightness and abdominal fullness for 8 months and massive pericardial effusion for 2 months. A large amount of pericardial effusion was found during the hospitalization of Gastroenterology. Then she was transferred to cardiology. Although infectious, tuberculous, and neoplastic pericardial effusions were excluded, there was still no diagnosis. The patients were examined by FDG-PETCT which considered IgG4-RD. After coming to our department, the results of the patient's laboratory tests showed that immunoglobulin subgroup IgG4 was 14.0 g/L. Then we performed a biopsy of the right submandibular gland. Pathological morphology and immunohistochemistry suggested IgG4-RD. Combined with level of IgG4, clinical, pathological and immunohistochemical results, we determined the final diagnosis of IgG4 related diseases. Then we gave glucocorticoid and immunosuppressant treatment. At the end, pericardial effusion was completely absorbed. As prednisone acetate was gradually reduced, no recurrence of the disease has been observed. Conclusion: Pericardial effusion can be the initial presentation in IgG4-RD. For patients with massive pericardial effusion of unknown cause, early detection of IgG4 is recommended, and PETCT may be helpful for obtaining the diagnosis.
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Doença Relacionada a Imunoglobulina G4 , Derrame Pericárdico , Feminino , Humanos , Idoso de 80 Anos ou mais , Doença Relacionada a Imunoglobulina G4/diagnóstico por imagem , Fluordesoxiglucose F18 , Derrame Pericárdico/diagnóstico por imagem , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada , Imunoglobulina GRESUMO
Process economy and dynamic controllability are critical for DMC/MeOH separation via the PSD process. In this paper, rigorous steady-state and dynamic simulations of atmospheric-pressurized process for DMC/MeOH separation with no, partial, and full heat integration have been carried out with Aspen Plus and Aspen Dynamics. Further investigations have been conducted into the economic design and dynamic controllability of the three neat systems. Simulation results indicated that: the separation process via full and partial heat integration provided TAC savings of 39.2 and 36.2%, respectively, compared to that of no heat integration; the non-heat-integrated system displays good dynamic performance, critical dynamic penalties were demonstrated for both partial and full heat integration processes, while the partial one exhibited a more robust control except for precisely maintaining XB2(DMC); a PCTC scheme with a CC/TC cascade control was proposed to precisely maintain the product concentration for the fully heat-integrated PSD process. A comparison of the economy between atmospheric-pressurized and pressurized-atmospheric sequences indicated that the former is more energy efficient. Further, a comparison of the economy between atmospheric-pressurized and pressurized-atmospheric sequences indicated that the former is more energy efficient. This study will provide new insights into the energy efficiency and has some implications for design and control of DMC/MeOH separation in the industrialization process.
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Matrix nanotopography plays an essential role in regulating cell behaviors including cell proliferation, differentiation, and migration. While studies on isolated single cell migration along the nanostructural orientation have been reported for various cell types, there remains a lack of understanding of how nanotopography regulates the behavior of collectively migrating cells during processes such as epithelial wound healing. We demonstrated that collective migration of epithelial cells was promoted on nanogratings perpendicular to, but not on those parallel to, the wound-healing axis. We further discovered that nanograting-modulated epithelial migration was dominated by the adhesion turnover process, which was Rho-associated protein kinase activity-dependent, and the lamellipodia protrusion at the cell leading edge, which was Rac1-GTPase activity-dependent. This work provides explanations to the distinct migration behavior of epithelial cells on nanogratings, and indicates that the effect of nanotopographic modulations on cell migration is cell-type dependent and involves complex mechanisms.
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Células Epiteliais , Cicatrização , Diferenciação Celular , Movimento Celular , Proliferação de CélulasRESUMO
Tunable In(OH)3-coupled Cu2O-derived hybrid catalysts are facilely synthesized to boost the selectivity and efficiency of the electrochemical CO2 reduction reaction (CO2RR). The maximum faradaic efficiency (FE) of 90.37% for CO production is achieved at -0.8 V versus reversible hydrogen electrode. The mechanistic discussion suggests that the composition-dependent synergistic effect results in the enhanced selectivity for CO on the hybrid catalyst. By increasing the concentration of the electrolyte, a dramatically enhanced current density of 40.17 mA cm-2 was achieved at -1.0 V in 0.7 M KHCO3. Furthermore, a KHCO3 electrolyte with high concentration promotes the selectivity of CO2RR over the low overpotential range. At a low overpotential of 290 mV, the increased FE for CO of 74.05% is obtained in 0.7 M KHCO3 as compared to that of 57.04% in 0.1 M KHCO3. Combining with the synergistic effect of the catalyst and the concentration effect of the electrolyte, the hybrid catalyst achieves high efficiency, high selectivity, and high stability for CO2RR.
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This paper describes the novel preparation of three kinds of nanofibers [poly(styrene-co-methacrylic acid), poly(styrene-co-p-styrene sulfonate), polystyrene] investigated as solid-phase extraction (SPE) sorbents to extract six compounds (nitrobenzene, 2-naphthol, benzene, n-butyl p-hydroxybenzoate, naphthalene, p-dichlorobenzene) in environmental water by high-performance liquid chromatography. Parameters affecting extraction efficiency were investigated in detail to explore the extraction mechanism of the nanofibers. Under optimized conditions, six compounds followed an excellent linear relationship in the range 10-5,000 ng mL(-1) with coefficients of determination (r (2)) greater than 0.99. The repeatability (expressed as relative standard deviations) was from 3.0 to 7.0%, corresponding to 2.0 mL of water samples at 25 and 500 ng mL(-1) spiked levels for the six compounds. The limits of detection varied from 0.01 to 0.15 ng mL(-1) (signal-to-noise ratio of 3). A comparison of the SPE using nanofibers as sorbents and the most commonly used octadecylsilica SPE cartridges was carried out in terms of absolute recovery, sensitivity, and reproducibility for the compounds investigated. Finally, the method was applied to four real water samples. The results highlighted the importance of functional groups, and the polarity of nanofibers in controlling sorption of target compounds, and clearly showed that the new method could be a viable and environmentally friendly technique for analyzing pollutants in environmental samples.
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Poluentes Ambientais/química , Nanopartículas/química , Nanotecnologia/métodos , Extração em Fase Sólida , Cromatografia Líquida de Alta Pressão/métodos , Monitoramento Ambiental/métodos , Desenho de Equipamento , Hidrocarbonetos/química , Concentração de Íons de Hidrogênio , Polímeros/química , Dióxido de Silício/química , Solventes/química , Espectrofotometria Infravermelho/métodos , Água/química , Poluentes Químicos da Água/análiseRESUMO
Given that physical restraints cause adverse effects to patients and nurses, their wide and frequent use has resulted in various clinical and ethical controversies in mental health services. The rate of physical restraints is significantly higher in China than in other countries. Some western and domestic media blamed relevant institutions for compromising the basic rights of psychiatric patients. Therefore, this study aims to collect and synthesize the relevant ethical evidence and to provide corresponding guidance for the nursing practice based on the current situation of the mainland of China. This study synthesized the ethical issue according to the ethical principle of autonomy, justice, beneficence, and non-maleficence. Given the current situation where the nursing workforce is limited and the mental health service is under-developed in China, applying physical restraints in the psychiatric inpatients to guarantee the safety of patients and nurses is crucial. In regard to respect the basic rights of psychiatric patients, it is recommended to protect the their autonomy, and eliminate the adverse effects of physical restraint. This goal would be achieved by ensuring the informed consent, providing humane care, and regulating the implementation of physical restraints.
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Endostar, a modified recombinant human endostatin, inhibits the growth of a variety of tumors by suppressing neovascularization. Vascular endothelial growth factor (VEGF) has an important role in malignant ascites formation. In order to determine whether Endostar can suppress the formation of ascites and prolong survival times, mouse models of malignant ascites were established using S180 and H22 tumor cells. The experimental mice were randomly divided into four groups: The three treatment groups received different doses of Endostar (4, 8 and 16 mg/kg), and the control group received 0.9% w/v NaCl. The volume of ascites, and the tumor cell, red blood cell (RBC), VEGF protein and mRNA content of the ascites was measured alongside the peritoneal permeability and the mouse survival time. In vitro analysis of cultured Endostar-treated S180 and H22 cells was also performed in order to examine cellular proliferation and the level of VEGF secreted protein and mRNA. The results revealed that Endostar suppressed the ascites volume, decreased the level of tumor cells, RBCs and VEGF in the ascites fluid, and lowered the permeability of the peritoneum. The tumor cells collected from the ascites in the Endostar-treated mice demonstrated a decrease in the expression of VEGF mRNA. The survival rates of the 8 and 16 mg/kg Endostar-treated mice were longer than those of the controls. The in vitro experiments revealed a significant inhibition of VEGF protein secretion and VEGF mRNA by Endostar, but no effect on cellular proliferation. In conclusion, Endostar lowers ascites production by downregulating VEGF expression, and may therefore be effective for the treatment of malignant ascites.
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OBJECTIVE: To investigate the effect of endostar in controlling ascites tumor formation in mice. METHODS: Mouse models bearing ascites tumors were established via intraperitoneal injection of H22 and S180 cell lines. Eighty-eight ICR mice were randomly assigned into 4 groups, namely the control group (0.9% normal saline) and 3 endostar groups with 8 mg/kg endostar administration daily, every other day or every two days. The peritoneal membrane permeability of the mice was assessed using Evan blue staining. The body weight curve of mice was drawn, and the cumulative ascites volume and number of red cells and tumor cells in the malignant ascites were determined. The survival of the mice was evaluated to assess the therapeutic effect of endostar. RESULTS: Compared with the control group, the mice receiving daily endostar injection showed obviously lower ascites accumulation and peritoneal capillary permeability (P<0.05) with reduced count of ascites tumor cells and red cells and tumor burden of the abdominal cavity. The mice with daily endostar injection also showed longer survival than the control group. CONCLUSIONS: Continuous intraperitoneal injection can be the optimal means for endostar administration for treatment of malignant ascites.
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Carcinoma de Ehrlich/irrigação sanguínea , Endostatinas/administração & dosagem , Endostatinas/farmacologia , Animais , Linhagem Celular Tumoral , Endostatinas/uso terapêutico , Feminino , Humanos , Camundongos , Camundongos Endogâmicos ICR , Neovascularização Patológica , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/farmacologia , Proteínas Recombinantes/uso terapêuticoRESUMO
Photonic crystal sensing materials have been validated that they are very sensitive to refractive index changes. Herein, three-dimensionally ordered macroporous (3DOM) (>50 nm) TiO2 inverse opal film has been fabricated by the self-assembly technique. Based on the TiO2 inverse opal film, the optical spectrometer was established for label-free immunosensor. The sensing performance of the 3DOM TiO2 was investigated using human IgG/goat anti-human IgG couple, which showed that the sensitivity of 3DOM TiO2 inverse opal film could reach to 1 microg mL(-1) (equivalent to 1.5 pg mm(-2)) of protein concentration detection limit. The 3DOM TiO2 inverse opal has a large internal surface area, low fluorescence background and unique optical properties. These characteristics indicated the feasibility of 3DOM TiO2 inverse opal in label-free immunoassay.
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Técnicas Biossensoriais/métodos , Imunoensaio/métodos , Titânio/química , Animais , Anticorpos Anti-Idiotípicos/análise , Anticorpos Anti-Idiotípicos/imunologia , Anticorpos Anti-Idiotípicos/metabolismo , Cabras/imunologia , Humanos , Imunoglobulina G/análise , Imunoglobulina G/imunologia , Imunoglobulina G/metabolismo , Óptica e Fotônica , Porosidade , Ligação Proteica , Coloração e Rotulagem , Especificidade por Substrato , Propriedades de Superfície , Titânio/metabolismoRESUMO
OBJECTIVE: To Compare the therapeutic effects of cryocareTM cryoablation, radiofrequency ablation(RFA), and and microwave coagulation (MCT) in rabbits with VX(2) liver cancer. METHODS: Forty-five rabbits with VX(2) liver cancer were randomly and equally allocated into 5 groups to receive treatment with cryocare cryoablation (group A), radiofrequency ablation (group B), microwave coagulation (group C), surgical resection (group D) and control group (group E), respectively. The residual tumor tissues and metastasis (intrahepatic, lung, abdominal lymphoid node, and abdominal implantation) were observed after the treatments, with also detection of soluble interleukin-2 receptor ( sIL-2R) and recording of the survival time of the rabbits. RESULTS: Significant differences were found in the occurrence of tumor residue (chi(2)=20.700, P=0.0000), intrahepatic metastasis (chi(2)=15.652, P=0.0004), and abdominal implantation tumor (chi(2)=13.894, P=0.0008) between the 5 groups, but not in lung and abdominal lymph node metastasis. sIL-2R levels differed significantly only after but not before the treatments (F=31.58, P=0.000) between groups A to D and group E (t=10.119, P=0.000). The treatments in groups A to D all resulted in prolonged survival of the rabbits as compared with the control (F=73.084, P=0.000), and cryocareTM cryoablation and surgical resection showed similarly better effect than RFA and MCT. CONCLUSION: Cryocare cryoablation can be more effective than RFA and MCT in reducing tumor residue and metastasis and prolonging the survival time of rabbits with VX(2) liver cancer, and RFA and MCT are comparable for their therapeutic effects.
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Técnicas de Ablação/métodos , Criocirurgia , Neoplasias Hepáticas/cirurgia , Micro-Ondas , Ondas de Rádio , Animais , Feminino , Neoplasias Hepáticas/metabolismo , Masculino , Coelhos , Receptores de Interleucina-2/metabolismo , Taxa de SobrevidaRESUMO
The study was aimed to investigate the expression of HLA class I molecules and MHC class I chain-related molecules A/B (MICA/MICB) in K562 and adriamycin (ADM)-resistant K562 cell lines (K562/AO2) and their effect on cytotoxicity of NK cells. Expression of HLA class I molecules and MICA/MICB on the surface of K562 and K562/AO2 cell lines were analyzed by flow cytometry. Cytotoxicity of NK cells (isolated from 3 healthy persons) against K562 and K562/AO2 cells were detected by LDH releasing assay at different effect-to-target cell ratios (E:T). In blocking experiments, anti-MHC class I monoclonal antibody (McAb) (W6/32, a pan anti-HLA class I antibody) and anti-MHC class I chain-related molecules McAb (BAMO-1, specifically against MICA and MICB) were added to the target cells at E:T of 10:1. The results showed that the expression of MHC class I chain-related molecules on K562 was higher than that on K562/AO2 (P=0.000), and HLA class I molecules were not detectable on both cells. Cytotoxicities of NK cells against K562 and K562/AO2 cells were (29.32 +/- 0.12)%, (45.33 +/- 0.78)%, (58.37 +/- 0.87)%, (72.37 +/- 0.96)% and (12.47 +/- 0.91)%, (24.36 +/- 1.11)%, (33.29 +/- 1.03)%, (53.87 +/- 1.27)% at E:T ratios of 5:1, 10:1, 20:1 and 30:1 respectively (P=0.000), the cytotoxicity of NK cells on K562 cells was significantly higher than that on K562/A02 cells at different E:T ratios. Blocking experiments confirmed that at E:T of 10:1 killing of NK cells against K562 and K562/AO2 cells was efficiently inhibited by BAMO-1, whereas W6/32 had no effect on K562 and K562/AO2 cells. It is concluded that the expression of MHC class I chain-related molecules on K562 and K562/AO2 cells is correlated with NK cell-mediated lysis. NK cells display higher cytotoxicity against parental K562 cells than multi-drug resistant K562/AO2 cells. Down-regulation of MICA/B in multi-drug resistant tumor cell lines leads to reduction of susceptibility to NK lysis.
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Citotoxicidade Imunológica/imunologia , Resistencia a Medicamentos Antineoplásicos/imunologia , Genes MHC Classe I/genética , Antígenos de Histocompatibilidade Classe I/imunologia , Células Matadoras Naturais/imunologia , Doxorrubicina/farmacologia , Humanos , Células K562RESUMO
OBJECTIVE: To observe the effect of thermotherapy on the intracellular adriamycin concentration and apoptosis of Raji cells in vitro. METHODS: The working concentration of adriamycin against Raji cells was determined with MTT assay. Raji cells were subjected to thermotherapy (at 40 degrees Celsius;, 41 degrees Celsius; or 42 degrees Celsius;) and chemotherapy with adriamycin alone or in conjunction, and the cell survival rates were determined 48 h after the treatment. The cell growth inhibition effect of the treatment was evaluated with MTT assay, and the apoptotic rates of Raji cells and alteration of intracellular adriamycin concentration were determined by flow cytometry. RESULTS: The IC(50) of adriamycin was defined as the working concentration in the experiment. Thermotherapy at 40, 41 and 42 degrees Celsius; for 60 min in conjuction with chemotherapy significantly inhibited the growth of Raji cells (P<0.01). The results of flow cytometry showed that thermotherapy and adriamycin chemotherapy, used either alone or in combination, significantly increased the apoptotic rates of Raji cells (P<0.01), and thermotherapy remarkably increased the intracellular concentration of adriamycin. CONCLUSION: Adriamycin chemotherapy combined thermotherapy for 60 min can increase the intracellular concentration of adriamycin and the apoptosis rates of Raji cells.
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Apoptose/efeitos dos fármacos , Doxorrubicina/farmacologia , Temperatura Alta , Antibióticos Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Terapia Combinada , Doxorrubicina/metabolismo , Citometria de Fluxo , Humanos , Hipertermia Induzida , Concentração Inibidora 50 , Espaço Intracelular/metabolismo , Linfoma/tratamento farmacológico , Linfoma/metabolismo , Linfoma/patologiaRESUMO
OBJECTIVE: To analyze the expression of NKG2D ligands on human nasopharyngeal carcinoma cell line CNE2 and the multidrug-resistant lin CNE2/DDP and investigate its impact on cytotoxicity of natural killer (NK) cells. METHODS: Expression of NKG2D ligands on the surface of CNE2 and CNE2/DDP cells was analyzed by flow cytometry, and their HLA genotypes, along with inhibitory killer cell immunoglobulin-like receptors (KIRs) expressed on NK cells from 5 healthy donors, were determined by PCR with sequence specific primers. Cytotoxicity of NK cells against CNE2 and CNE2/DDP cells was evaluated by LDH-releasing assay at different effector-to-target ratios (E:T). In blocking experiments, different monoclonal antibodies (mAb) were added to the target cells at the E:T of 20:1 ratio. RESULTS: The HLA genotypes of CNE2 and CNE2/DDP cells were A2, 24, B18, 35, Cw4, 7, and the inhibitory KIR genotypes of the 5 healthy donors were KIR2DL1, KIR2DL3, KIR3DL1, and KIR3DL2, mismatched with the HLA -class I molecules expressed by the CNE2 and CNE2/DDP cells. The expression of MHC class I chain-related proteins A and B (MICA and MICB) on CNE2 cells was higher than that on CNE2/DDP cells (P<0.01), and ULBP1 and ULBP3 were not detectable. NK cells displayed highly in vitro cytotoxicity against CNE2 and CNE2/DDP cells with a mean cell lysis rate of (10.50-/+2.17)%, (4.98-/+0.95)%; (27.68-/+1.47) %, (15.48-/+2.10) %; (36.99-/+3.13) %, (28.46-/+4.30) %; (55.00-/+2.20) %, (40.95-/+2.21) %, respectively, corresponding to the E:T ratios of 5:1, 10:1, 20:1, and 30:1 (P<0.01). Blocking experiments confirmed that killing of CNE2 and CNE2/DDP cells by NK cells was efficiently inhibited by anti-MICA, anti-MICB, and anti-ULBP2 mAbs, whereas anti-ULBP1 and anti-ULBP3 mAbs had no effects on the cytotoxicity of the NK cells. CONCLUSION: Expression of NKG2D ligands on CNE2 and CNE2/DDP cells is correlated with NK cell-mediated lysis, and NK cells display higher cytotoxicity against parental CNE2 cells than the multidrug-resistant CNE2/DDP cells.
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Antígenos de Histocompatibilidade Classe I/metabolismo , Células Matadoras Naturais/imunologia , Subfamília K de Receptores Semelhantes a Lectina de Células NK/metabolismo , Anticorpos/farmacologia , Linhagem Celular Tumoral , Citotoxicidade Imunológica/efeitos dos fármacos , Citotoxicidade Imunológica/imunologia , Resistência a Múltiplos Medicamentos , Citometria de Fluxo , Genótipo , Antígenos HLA/genética , Antígenos de Histocompatibilidade Classe I/imunologia , Humanos , Subfamília K de Receptores Semelhantes a Lectina de Células NK/imunologia , Neoplasias Nasofaríngeas/imunologia , Neoplasias Nasofaríngeas/metabolismo , Neoplasias Nasofaríngeas/patologia , Reação em Cadeia da Polimerase , Receptores KIR/genéticaRESUMO
OBJECTIVE: To study the effect of adriamycin combined with hyperthermia on tumor formation and growth of human B lymphoma cell line (Raji) in nude mice. METHODS: Twenty-four BALB/C nude mice were divided into control group (37 degrees celsius;), chemotherapy group (37 degrees celsius;+ADM), hyperthermia group (42 degrees celsius;) and thermochemotherapy group (42 degrees celsius;+ADM), and in each mouse, 5 x 10(6) Raji cells were injected subcutaneously. The time and rate of tumor formation were observed, the tumor diameter was measured every 3 days and the tumor volume calculated to obtain the growth curves of the tumors. Three weeks after tumor formation, all the mice were executed to observe the histopathological changes of the tumor with HE staining. RESULTS: The time of tumor formation in the control, chemotherapy, hyperthermia and thermochemotherapy groups were 11.2-/+1.7, 20.2-/+2.3, 15.3-/+1.6 and 23.8-/+1.7 days, respectively. Three weeks after tumor formation, the average weight of the tumors were 3.33-/+0.57, 2.26-/+0.28, 2.76-/+0. 26 and 1.73-/+0. 33 g, respectively, and the tumor growth inhibition rate was 48.0% in the thermochemotherapy group. CONCLUSION: Thermochemotherapy can be effective in inhibiting tumor growth and provides an alternative of adjuvant therapy for malignant B cell lymphoma.
Assuntos
Transformação Celular Neoplásica , Doxorrubicina/farmacologia , Hipertermia Induzida , Linfoma de Células B/patologia , Animais , Antígenos CD20/metabolismo , Linhagem Celular Tumoral , Terapia Combinada , Doxorrubicina/uso terapêutico , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Antígenos Comuns de Leucócito/metabolismo , Linfoma de Células B/tratamento farmacológico , Linfoma de Células B/genética , Linfoma de Células B/terapia , Masculino , Camundongos , Camundongos Nus , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
This study was purposed to investigate the inhibitory effect, apoptosis, Bcl-2 and P-gp expression of K562/AO2 cells by hyperthermia combined with adriamycin. The working concentration of adriamycin against K562/AO2 was determined by MTT assay. The hyperthermia and chemotherapy were used alone or in combination, then the cell survival rate was detected at 48 hours. The inhibitory effect was evaluated by MTT assay. The apoptosis rate, Bcl-2 and P-gp expression of K562/AO2 were determined by flow cytometry. The concentration of adriamycin in the experiment was defined as its IC(50) at 48 hours action. The results indicated that the hyperthermia at 40, 41 and 42 degrees C for 60 minutes showed obvious inhibitory effect on K562/AO2 cells (p < 0.01). Adriamycin chemotherapy combined with hyperthermia showed more obvious inhibitory effect on K562/AO2. According to flow cytometric analysis, the hyperthermia and adriamycin used alone or in combination could obviously increase the apoptosis rate and down-regulate Bcl-2 and P-gp expression of K562/AO2 cells (p < 0.01). It is concluded that the adriamycin chemotherapy combined with hyperthermia for 60 minutes shows obvious inhibitory effect on K562/AO2 cells, which increases the apoptosis rate and down-regulates expression of Bcl-2 and P-gp.