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The aim of this study is to investigate certain genetic features of intrahepatic cholangiocarcinoma (ICCA). A total of 12 eligible ICCA patients were enrolled, and tumor tissues from the patients were subjected to next-generation sequencing of a multi-genes panel. Tumor mutation burden (TMB), mutated genes, copy number variants (CNVs), and pathway enrichment analysis were performed. The median TMB was 2.76 Mutation/Mb (range, 0-36.62 Mutation/Mb) in ICCA patients. The top two most commonly mutated genes in ICCA were KRAS (33%) and TP53 (25%). The co-mutations of KRAS and TP53 were 16.7% (2/12) in ICCA patients. Notably, patient P6 with the highest TMB did not have KRAS and TP53 mutations. Additionally, TP53 and/or KRAS alterations were significantly associated with poor progression-free survival than those with wild type (1.4 months vs 18 months). DNA damage repair and homologs recombinant repair deficiencies were significantly associated with high TMB in ICCA cases. In conclusion, we found that certain genetic mutations of TP53 and KRAS could predict poor prognosis in ICCA patients.
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Lithium (Li) metal batteries are among the most promising devices for high energy storage applications but suffer from severe and irregular Li dendrite growth. Here, it is demonstrated that the issue can be well tackled by precisely designing the leaf-like membrane with hierarchical structure and exquisite fluidic channels. As a proof of concept, plant leaf-inspired membrane (PLIM) separators are prepared using natural attapulgite nanorods. The PLIM separators feature super-electrolyte-philicity, high thermal stability and high ion-selectivity. Thus, the separators can guide uniform and directed Li growth on the Li anode. The Li//PLIM//Li cell with limited Li anode shows high Coulombic efficiency and cycling stability over 1500 h with small overpotential and interface impedance. The Li//PLIM//S battery exhibits high initial capacity (1352 mAh g-1 ), cycling stability (0.019% capacity decay per cycle at 1 C over 500 cycles), rate performance (673 mAh g-1 at 4 C), and high operating temperature (65 °C). The separators can also effectively improve reversibility and cycling stability of the Li/Li cell and Li//LFP battery with carbonate-based electrolyte. As such, this work provides fresh insights into the design of bioinspired separators for dendrite-free metal batteries.
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Non-enzymatic electrochemical sensors with significant advantages of high sensitivity, long-term stability, and excellent reproducibility, are one promising technology to solve many challenges, such as the detection of toxic substances and viruses. Among various materials, perovskite oxides have become a promising candidate for use in non-enzymatic electrochemical sensors because of their low cost, flexible structure, and high intrinsic catalytic activity. A comprehensive overview of the recent advances in perovskite oxides for non-enzymatic electrochemical sensors is provided, which includes the synthesis methods of nanostructured perovskites and the electrocatalytic mechanisms of perovskite catalysts. The better sensing performance of perovskite oxides is mainly due to the lattice O vacancies and superoxide oxygen ions (O22-/O-), which are generated by the transfer of lattice oxygen to adsorbed -OH and have performed excellent properties suitable for electrooxidation of analytes. However, the limited electron transfer kinetics, stability, and selectivity of perovskite oxides alone make perovskite oxides far from ready for scientific development. Therefore, composites of perovskite oxides with other materials like graphitic carbon, metals, metal compounds, conducting organics, and biomolecules are summarized. Furthermore, a brief section describing the future challenges and the corresponding recommendation is presented in this review.
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Without insight into the correlation between the structure and properties, anion exchange membranes (AEMs) for fuel cells are developed usually using the empirical trial and error method or simulation methods. Here, a virtual module compound enumeration screening (V-MCES) approach, which does not require the establishment of expensive training databases and can search the chemical space containing more than 4.2×105 candidates was proposed. The accuracy of the V-MCES model was considerably improved when the model was combined with supervised learning for the feature selection of molecular descriptors. Techniques from V-MCES, correlating the molecular structures of the AEMs with the predicted chemical stability, generated a ranking list of potential high stability AEMs. Under the guidance of V-MCES, highly stable AEMs were synthesized. With understanding of AEM structure and performance by machine learning, AEM science may enter a new era of unprecedented levels of architectural design.
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The harmful effects on the human body from p-phenylenediamine (PPD) in hair dyes can cause allergies and even cancer. Therefore, it is particularly important to accurately control and detect the content of PPD in our daily products and environment. Here, a small amount of non-metallic elemental P doped in perovskite oxide of SrCoO3-δ (SC) forms a good catalytic material, SrCo0.95P0.05O3-δ (SCP), for PPD detection. The improved performance compared with that of the parent SC can be attributed to three contributing factors, including a larger amount of highly oxidative oxygen species O22-/O-, better electrical conductivity, and more active sites on the P5+-oxygen bonds of SCP. Moreover, the lattice oxygen mechanism (LOM) with highly active species of lattice O vacancies and adsorbed -OO for electrocatalytic oxidation of PPD by the SCP/GCE (glass carbon electrode) sensor is proposed in our work. More importantly, the SCP/GCE sensor exhibits good stability, a low limit of detection, and high reliability (error < 5.78%) towards PPD determination in real samples of hair dyes, suggesting the substantial research potential for practical applications.
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BACKGROUND: The source and availability of cells for tissue engineering in large scale research or clinical trials requires special attention. We propose the idea of applying rabbit umbilical cord mesenchymal stem cells for this purpose. METHODS: Here, the structure of the rabbit umbilical cord was analyzed and compared to that of human umbilical cord, both macroscopically and histologically. Next, we isolated, cultured and identified the proliferative activity and immunological characteristics of rabbit umbilical cord mesenchymal stem cells in vitro using mixed lymphocyte reaction, flow cytometry and an enzyme-linked immunosorbent assay. Furthermore, we evaluated the effects of biphasic calcium phosphate ceramic scaffolds seeded with rabbit umbilical cord mesenchymal stem cells in rat cranial defect models using multiple techniques, including radiological, histological and immunohistochemistry. RESULTS: In vitro studies demonstated a high level of proliferation and multi-lineage differentiation potential in rabbit umbilical cord mesenchymal stem cells. Rabbit umbilical cord mesenchymal stem cells exibited low immunogenicity properties and immune suppression capability with respect to both the allogeneic and xenogeneic immune response. The results of the in vivo study showed that rabbit umbilical cord mesenchymal stem cells could promote osteogenesis in heterogeneous hosts. CONCLUSIONS: The rabbit umbilical cord mesenchymal stem cells may be a new source for tissue engineering.
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Diferenciação Celular , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Engenharia Tecidual , Cordão Umbilical/citologia , Animais , Biomarcadores , Linhagem da Célula/genética , Proliferação de Células , Células Cultivadas , Citocinas/metabolismo , Humanos , Imunomodulação , Imunofenotipagem , Transplante de Células-Tronco Mesenquimais , Osteogênese/genética , Coelhos , Engenharia Tecidual/métodosRESUMO
Adipogenesis participates in many physiological and pathological processes, such as obesity and diabetes, and is regulated by a series of precise molecular events. However, the molecules involved in this regulation have not been fully characterized. In this study, we identified a long noncoding (lnc)RNA, lncRNA-Adi, which is highly expressed in adipose tissue-derived stromal cells (ADSCs) that are differentiating into adipocytes. Knockdown of lncRNA-Adi impaired the adipogenic differentiation ability of ADSCs. Moreover, lncRNA-Adi was found to interact with microRNA (miR)-449a to enhance the expression of cyclin-dependent kinase (CDK)6 during adipogenesis. The mechanism by which lncRNA-Adi regulates adipogenesis was determined to involve an lncRNA-Adi-miR-449a interaction that competes with the CDK6 3' untranslated region to increase CDK6 translation and activate the pRb-E2F1 pathway to promote adipogenesis. These findings provide valuable information and a new study angle to search for therapeutic targets against metabolic disorders such as obesity and diabetes.
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Adipogenia/genética , RNA Longo não Codificante/metabolismo , Tecido Adiposo/citologia , Animais , Sequência de Bases , Quinase 6 Dependente de Ciclina/metabolismo , Fator de Transcrição E2F1/metabolismo , Feminino , Regulação da Expressão Gênica , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Longo não Codificante/genética , Ratos Wistar , Proteína do Retinoblastoma/metabolismo , Células Estromais/citologia , Células Estromais/metabolismo , Transcriptoma/genéticaRESUMO
TMC-incorporated carbon nitride (CN) with hexagonal and quadrangle honeycomb-like structure and having periodic lattice defects linked by -CONH- bond was synthesized through combining the high calcination with the chemical condensation of melamine and 1,3,5-benzenetricarbonyl trichloride. The obtained CN has a tri-s-triazine ring and benzene ring skeleton, which makes it have excellent mechanical and thermal stability. The BET specific surface area was enhanced to about 125.6 m2/g, and the mean pore size is about 3.43 nm. This CN exhibited an excellent adsorption-enhanced photocatalytic performance.
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RATIONALE: Ectopic variceal bleeding due to hepaticojejunostomy (HJ) is unusual and difficult to manage. Reports on the use of side-to-side splenorenal shunt for severe bleeding from varices at HJ anastomosis are lacking. PATIENT CONCERNS: A 43-year-old man was admitted to our hospital with repeated episodes of hematemesis. He has a history of right hemihepatectomy with HJ reconstruction to the left hepatic duct for hilar cholangiocarcinoma. Two years after surgery, he presented with repeated episodes of hematemesis and underwent blood transfusion. DIAGNOSES: Imaging tests and endoscopic investigation failed to identify the bleeding source. When conservative management failed to control his bleeding, he underwent emergency laparotomy, which revealed hemorrhage from ectopic varices at the HJ anastomosis. INTERVENTIONS: To arrest the bleeding, a side-to-side venovenal anastomosis was created between the splenic and left renal veins to form a shunt for decompression of the varices at the HJ anastomosis. OUTCOMES: After the surgery, the patient's symptoms ceased, and a no bleeding in the digestive tract was noted at 2-year follow-up. LESSONS: The present patient is the first reported case of unusual bleeding from HJ controlled by a side-to-side splenorenal shunt. We believe this approach is a useful and effective surgical treatment for severe bleeding from varices at the HJ anastomosis.
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Jejuno/cirurgia , Fígado/cirurgia , Hemorragia Pós-Operatória/cirurgia , Derivação Esplenorrenal Cirúrgica/métodos , Adulto , Humanos , MasculinoRESUMO
New poly(N-isopropylacrylamide) brushes grafted with ZrO2 (PNIPAAm-g-ZrO2) composite membranes, which had been prepared in our laboratory, were used for humic acid (HA) removal. We found that the fluxes associated with such membranes, when compared to those obtained from unmodified ZrO2 membranes, declined slightly at both 25 °C and 35 °C. The PNIPAAm-g-ZrO2 membrane achieved a high rejection, of 98.0%, at a suitable steady flux of 111.9 L m(-2) h(-1) at 25 °C. This membrane exhibited good anti-fouling properties as well as improved membrane performance during filtration of HA. The important role of pH and Ca(2+) concentration in HA removal was also investigated. Lower adsorption fouling and a higher rejection were obtained at higher pH levels. The Ca(2+) ions reduced the electrostatic exclusion and played a cross-linking role between HA and the PNIPAAm-g-ZrO2 membrane surface. Fouling was severe in the presence of Ca(2+). These tests led to the development of an environment-friendly membrane cleaning method, by means of temperature-change water elution around LCST of PNIPAAm-brushes. After the alternate temperature-change (25 °C/35 °C) cleaning, a flux recovery of 98.2% was obtained for the PNIPAAm-g-ZrO2 membrane. Moreover, after four repeated experiments, the anti-fouling and easy-cleaning properties were still maintained. It is implied that PNIPAAm-brushes were firmly "stuck" to the membrane surface, and could not easily be removed by water cleaning or HA filtration. The PNIPAAm-g-ZrO2 membranes exhibited good stability and great potential for HA removal.
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Acrilamidas/química , Substâncias Húmicas/análise , Membranas Artificiais , Polímeros/química , Temperatura , Zircônio/química , Resinas Acrílicas , Incrustação Biológica/prevenção & controle , Cálcio/química , Filtração , Concentração de Íons de Hidrogênio , Microscopia Eletrônica de Varredura , Padrões de Referência , Soluções , Eletricidade Estática , Água/químicaRESUMO
Polyacrylamide/attapulgite (PAM/ATP) was prepared by the solution polymerization of acrylamide (AM) onto gamma-methacryloxypropyl trimethoxy silane (KH-570)-modified attapulgite (ATP). PAM/ATP was characterized using Fourier transform infrared (FTIR) and X-ray photoelectron spectroscopy (XPS). The effects of contact time, adsorbent dosage, and pH of the initial solution on the adsorption capacities for Hg(2+) were investigated. The adsorption process was rapid; 88% of adsorption occurred within 5 min and equilibrium was achieved at around 40 min. The equilibrium data fitted the Langmuir sorption isotherms well, and the maximum adsorption capacity of Hg(2+) onto PAM/ATP was found to be 192.5 mg g(-1). The adsorption kinetics of PAM/ATP fitted a pseudo-second-order kinetic model. Our results suggest that chemisorption processes could be the rate-limiting steps in the process of Hg(2+) adsorption. Hg(2+) adsorbed onto PAM/ATP could be effectively desorbed in hot acetic acid solution, and the adsorption capacity of the regenerated adsorbents could still be maintained at 95% by the sixth cycle.
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Resinas Acrílicas/química , Compostos de Magnésio/química , Mercúrio/análise , Mercúrio/isolamento & purificação , Metacrilatos/química , Silanos/química , Compostos de Silício/química , Poluentes Químicos da Água/análise , Purificação da Água/métodos , Adsorção , Concentração de Íons de Hidrogênio , Cinética , Espectrometria por Raios X , Espectroscopia de Infravermelho com Transformada de Fourier , Fatores de Tempo , Eliminação de Resíduos Líquidos/métodos , Poluentes Químicos da Água/isolamento & purificaçãoRESUMO
The study was aimed to investigate the influence of interferon alpha (IFN-alpha) on the expressions of Fas and Fas ligand (FasL) in dendritic cells (DCs) from patients with chronic myeloid leukemia (CML). In addition to adding stem cell factor (SCF), granulocyte-macrophage colony-stimulating factor (GM-CSF), tumor necrosis factor alpha (TNF-alpha) and interleukin 4 (IL-4), the IFN-alpha was added to the serum-free medium for DCs. After culturing for 10 - 14 days, cell phenotype and percentage of Ph(1) chromosome were detected by different methods. The expression of Fas or FasL on CML-DCs and cell cycle of DCs labeled with propidium iodine (PI) were measured by flow cytometry. The concentration of sFas in supernatants was analyzed by enzyme-linked immunosorbent assay (ELISA). The results indicated that the expression of co-stimulatory molecules were improved significantly while the percentages of Ph(1) positive cells decreased. The level of Fas on cells was up-regulated and the concentration of sFas decreased. However, the expression of FasL was negative. The ratio of apoptosis rose gradually while the concentration of IFN-alpha increased. It is concluded that IFN-alpha can accelerate the apoptosis of Ph(1) positive cells through Fas/FasL pathway, so the number of Ph(1) negative cells increases relatively.
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Células Dendríticas/metabolismo , Proteína Ligante Fas/metabolismo , Interferon-alfa/farmacologia , Leucemia Mielogênica Crônica BCR-ABL Positiva/metabolismo , Receptor fas/metabolismo , Adolescente , Adulto , Apoptose/efeitos dos fármacos , Células Cultivadas , Criança , Meios de Cultura/farmacologia , Células Dendríticas/citologia , Proteína Ligante Fas/genética , Feminino , Humanos , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Masculino , Pessoa de Meia-Idade , Cromossomo Filadélfia , Adulto Jovem , Receptor fas/genéticaRESUMO
This study was aimed to investigate the influences of interferonalpha (IFN-alpha) on expressions of CCR7, interleukin10 (IL-10) and IL-12p70 in dendritic cells (DCs) from patients with chronic myeloid leukemia (CML). In addition to stem cell factor (SCF), granulocyte-macrophage colony stimulating factor (GM-CSF), tumor necrosis factor-alpha (TNF-alpha) and IL-4, IFN-alpha was added to the serum-free medium of DCs. After culture for 10-14 days, phenotypes and function of CML-DCs were evaluated respectively by flow cytometry and methyl thiazolyl tetrazolium (MTT) assay. Chromosome of DCs was analyzed by displaying G banding assay. The concentrations of IL-10 and IL-12P70 in supernatants were evaluated by enzyme-linked immunosorbent assay (ELISA). The results showed that the expressions of CD40, CD83, CD86 and CCR7 and the OD value in allogeneic mixed-lymphocyte reaction (MLR) in group with IFN-alpha (300 U/ml) were twice as high as those in group without IFN-alpha. The percentage of Ph1 positive cells and concentrations of IL-10 and IL-12 P70 were reduced in group with IFN-alpha. It is concluded that the defective phenotypes and functions of CML-DCs can be recruited partly by IFN-alpha. The mechanism may lie in the facts that expression of CCR7 and co-stimulatory molecules is promoted and the inhibitory effect of IL-10 on CML-DCs is relieved partly through the regulation of IFN-alpha.
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Células Dendríticas/citologia , Interferon-alfa/farmacologia , Interleucina-10/metabolismo , Leucemia Mielogênica Crônica BCR-ABL Positiva/imunologia , Receptores CCR7/metabolismo , Células Cultivadas , Humanos , Interleucina-10/genética , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Cromossomo Filadélfia/efeitos dos fármacos , Receptores CCR7/genéticaRESUMO
OBJECTIVE: To survey the etiology of sensorineural hearing loss with unknown reason and the incidence of the mutation of SLC26A4 2168A > G in Henan province. METHOD: The evaluation of hearing loss, etiologic survey, the molecular genetic analysis and temporal bone CT examination for genes common to hereditary hearing disorders were performed in 95 hearing-impaired patients in Henan province. RESULT: In the deafness group, the incidence of large vestibular aqueduct syndrome (LVAS) which correlates with SLC26A4 2168A > G is 6.32%. The incidence of the gene diagnosis conformed to the clinical one is 83.3%. CONCLUSION: There is a high incidence of SLC26A4 2168 A > G mutation in sensorineural hearing loss with unknown reason. Molecular genetic screening for these mutations and genetic counseling are effective methods to prevent the occurrence of hereditary hearing loss.
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Perda Auditiva Neurossensorial/genética , Proteínas de Membrana Transportadoras/genética , Mutação , Sequência de Bases , China/epidemiologia , Testes Genéticos , Perda Auditiva Neurossensorial/epidemiologia , Humanos , Transportadores de Sulfato , Aqueduto VestibularRESUMO
OBJECTIVE: To investigate the available operation method of special type tracheobronchial foreign bodies removal. METHOD: Sixty one patients of special type tracheobronchial foreign bodies were extracted with some special instruments and methods depend on foreign body types. The features of operation on different kinds of foreign bodies were analyzed. RESULT: All patients were operated successfully by using foreign body hook (31 cases), pencil or writing screw brush extracted (11 cases), foreign body cage (13 cases), slender inverse claw esophagus foreign body clamp (3 cases) and so on. No complications and died cases were found. Only 5 patients were undertaken trachea incision before operation. CONCLUSION: Suitable instruments and technique chose for every individual cases is the key to successful operation.
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Brônquios , Corpos Estranhos/cirurgia , Traqueia , Adolescente , Adulto , Idoso , Broncoscopia/métodos , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Resultado do Tratamento , Adulto JovemAssuntos
Corpos Estranhos/cirurgia , Telas Cirúrgicas , Adolescente , Adulto , Idoso , Brônquios , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Traqueia , Adulto JovemRESUMO
To study the influence of IFN-alpha on function of CML-DC cultured in vitro and expression of chemokine and its chemokine receptor, bone marrow mononuclear cells from 13 CML patients were cultured in the fetal calf serum culture system supplemented with rhSCF, rhFlt-3L for expansion system, and adding rhGM-CSF, rhTNF-alpha, rhIL-4, with or without rhIFN-alpha to induce DCs. After incubation for two weeks, the phenotypes of CML-DC were analyzed by direct immunofluorescence and flow cytometry. The concentration of MIP-3beta expressed by CML-DC in the supernatant were analyzed by ELISA. The proliferative ability of T cells from healthy volunteers stimulated by CML-DCs were measured by MTT assay. The results showed that expression of CD86, CD83, CD40, MHC-I class molecules, CCR7, the concentration of MIP-3beta expressed by CML-DC, and the proliferative ability of T cells stimulated by CML-DCs in IFN-alpha group were all significantly higher than that in control group (P < 0.01). It is concluded that the immunophenotype of CML-DCs can be partially changed by IFN-alpha to accelerate the maturation of CML-DCs, enhance the capacity of CML-DCs, and stimulate allogeneic T lymphocyte proliferation.
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Quimiocinas/biossíntese , Células Dendríticas/efeitos dos fármacos , Interferon-alfa/farmacologia , Leucemia Mielogênica Crônica BCR-ABL Positiva/sangue , Receptores de Quimiocinas/biossíntese , Adulto , Idoso , Antígenos CD/análise , Antígeno B7-2/análise , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/metabolismo , Células da Medula Óssea/patologia , Antígenos CD40/análise , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Células Dendríticas/metabolismo , Células Dendríticas/patologia , Feminino , Citometria de Fluxo , Técnica Direta de Fluorescência para Anticorpo , Humanos , Imunoglobulinas/análise , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Leucócitos Mononucleares/patologia , Masculino , Glicoproteínas de Membrana/análise , Pessoa de Meia-Idade , Antígeno CD83RESUMO
The study was aimed to investigate the extensive amplification and the cytotoxicity of dendritic cells (DC) derived from chronic myeloid leukemia cells. DC were cultured in two steps: firstly, extensive amplification in primary culture of CD34(+) or mononuclear cells isolated from CML patients' bone marrow and peripheral blood with rhFlt3-L and rhTPO for 7 days; secondly, inducing culture of DC with rhGM-CSF, rhTNF and rhIL-4 for 14 days. A system inducing DC directly were established for comparison. DC were identified by immunophenotype with flow cytometry, chromosome analysis by displaying G banding and electric microscopy analysis. The function of stimulating T cells proliferation and cytotoxicity of CML cells were confirmed through MTT assay. The results showed that after first extensive amplification in primary culture with rhFlt3-L and rhTPO for 7 days, CD34(+) cells had a total cell number with (77 +/- 5) fold expansion, and DC were (39 +/- 8)% of total cell respectively after induction culture of DC with rhGM-CSF, rhTNF and rhIL-4 for 14 days. Both the amplification of cell number and yield of DC were higher than the system without extensively culture (P < 0.01). Such DC could stimulate T cells to proliferate and kill leukemia cells finally. In conclusion, two-step culture method can obviously improve the cell number of DC required, that is better than inducing them directly. DC derived from CML cells induce the generation of anti-leukemia immunization.
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Antígenos CD34/imunologia , Células Dendríticas/imunologia , Leucemia Mielogênica Crônica BCR-ABL Positiva/imunologia , Adolescente , Adulto , Idoso , Proliferação de Células , Células Cultivadas , Criança , Citotoxicidade Imunológica , Células Dendríticas/patologia , Feminino , Citometria de Fluxo , Humanos , Imunofenotipagem , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Masculino , Pessoa de Meia-Idade , Linfócitos T Citotóxicos/imunologiaRESUMO
OBJECTIVE: To study the effect of Ar(+) laser on human vas deferens and to compare the effects of using different radiation levels with varying thickness of tissue and varying levels of injury. METHODS: After initial tests on animals, four human scrotums were opened and treated directly with Ar(+) laser radiation. Then 58 human individual scrotums were treated with radiation by the method of trans-skin puncture. The rate of sperm reduction and elimination was tested. RESULTS: In 60 cases, the sperms were found to be eliminated completely after six months of radiation treatment. In 2 cases the sperms were found not to be eliminated completely due to the insufficient radiation. CONCLUSION: Ar(+) laser is one of the best forms of radiation for coagulation of vas deferens. It can be used to coagulate vas deferens without any complications or sequelae.
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Fotocoagulação a Laser , Esterilização Reprodutiva/métodos , Adulto , Seguimentos , Humanos , Masculino , Ducto Deferente/cirurgiaRESUMO
OBJECTIVE: To establish a serum-free culture system of dendritic cells (DCs) from chronic myeloid leukemia (CML) cells so that DCs vaccine may be applied to the adoptive immunotherapy of CML in the near future. METHODS: Fetal calf serum, serum-free medium and autologous serum were used for culture of DCs. The usage of cytokines was classified into two groups: group A (stem cell factor, granulocyte/macrophage colony-stimulating-factor, tumor necrosis factor-alpha and interleukin-4) and group B (granulocyte/macrophage colony-stimulating-factor, tumor necrosis factor-alpha and interleukin-4). The phenotypes of DCs were analyzed by using indirect immunofluorescence and flow cytometry. Mixed leukocyte responses were performed by methyl thiazolyl tetrazolium (MTT) assay. Chromosome analysis of DCs can be achieved by displaying G banding. T cells from CML patients were stimulated with autologous DCs and T-cell cytotoxicity was measured by (MTT) assay. RESULTS: CD34(+) cells or mononuclear cells were obtained from peripheral blood or bone marrow samples of eight patients of chronic-phase CML. Group A of serum-free medium was better than group B in expansion of total cell numbers and the rate of DCs. These results of serum-free medium were not significantly different from those of fetal calf serum medium, but the results of autologous serum medium were inferior to two groups above. The expression of major histocompatibility complex class II antigen on the surface of DCs was notable (> 50%), but the expression of CD83 and the costimulatory molecules CD86 was not noticeable (10% - 50%). Although CD1a(+)/CD14(-) DCs were potent stimulators of allogeneic lymphocytes, expansion of T cells from normal volunteers were not significant (average 27.2 fold at DCs: T cells ratio of 1:10). At day 12, CD1a(+) cells from three patients were studied by displaying G banding and Ph(+) cells in these populations were 100%, 98% and 60%, respectively. At an effector: target ratio of 40:1, 32% to 45% cytotoxicity was noted with DC-stimulated T cells against autologous leukemia cells. CONCLUSIONS: A stable serum-free culture system of CML-DCs was established. The expression of CD83 and CD86 on the surface of CML-DCs and DCs' potent stimulation of allogeneic lymphocytes were not notable. DCs in CML patients can be derived from the malignant clone and these malignant DCs could induce anti-leukemic reactivity in autologous T lymphocytes without the necessity for additional exogenous antigens.