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A survey was conducted on the application status of "Internet plus" technology in maternal and child health management in 31 provincial maternal and child health care institutions through the national comprehensive information platform for maternal and child health. 19 provincial institutions had realized one or more maternal and child health management and service functions under their jurisdiction through the regional health information platform, accounting for 61%. Among thirteen functions of management and service achieved in the regional health information platform, the top four were maternal system management (39%), high-risk maternal management (29%), high-risk newborn management (26%) and children system management (26%). Most functions were better in the eastern regions than those in the central and western regions. 15 provinces had established maternal and child health information platforms. 52% of provincial maternal and child health institutions provided telemedicine services, which were mainly for remote consultation. The main cooperative institutions of telemedicine services were subordinate health care institutions (39%).
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Serviços de Saúde da Criança , Atenção à Saúde , Internet , Serviços de Saúde Materna , Criança , China , Feminino , Humanos , Recém-Nascido , TelemedicinaRESUMO
OBJECTIVE: To explore the effect of breastfeeding promotion strategies on neonatal clinical outcomes of preterm infants during hospitalization in the neonatal intensive care unit (NICU). METHODS: We developed breastfeeding promotion strategies, including the establishment of a multidisciplinary breastfeeding steering team, breastfeeding support of families and society, family-integrated care, kangaroo mother care, donor human milk bank, and so on. Preterm infants meeting the inclusion standard, less than 32 weeks gestational age, who were admitted to NICU from November 2015 to February 2017 were enrolled, and the eligible infants were divided into two groups (control group and intervention group) before and after policy implementation. The data of preterm infants including breastfeeding related outcomes (time to initiation of enteral feeding, time to initiation of breastfeeding, time to achieve full breastfeeding, time to achieve full enteral feeding and rate of breastfeeding), growth (extrauterine growth restriction) and complications were compared between the two groups. RESULTS: One hundred and twenty-three preterm infants were enrolled, including 61 in the control group and 62 in the intervention group. There were no significant differences in gender, gestational age, birth weight, intrauterine growth retardation (IUGR) and admission disease status between the two groups (P>0.05). Compared with the control group, there were significantly earlier time to initiation of enteral feeding [15.37 (10.00, 22.13) h vs. 20.25 (12.88, 26.33) h, P<0.01], time to achieve full breastfeeding [91.00 (69.75, 103.00) h vs. 94.00 (80.37, 118.75) h, P=0.04], and time to achieve full enteral feeding [12 (11, 15) d vs. 14 (12, 18) d, P<0.01] in the intervention group. Otherwise, there were no significant differences in time to initiation of breastfeeding, hospital stay, extrauterine growth restriction (EUGR) occurance rate of weight, the rate of breastfeeding, motality, and the incidence of complications including feeding intolerance, neonatal necrotizing enterocolitis (NEC), bronchopulmonary dysplasia (BPD) and retinopathy of prematurity (ROP) (P>0.05). CONCLUSION: The breastfeeding promotion strategie was a quality improvement of ordinary breastfeeding protocol. It had significantly reduced time to initiation of enteral feeding, time to achieve full breastfeeding and time to achieve full enteral feeding for preterm infants in NICU. Further research is needed to confirm whether the strategies can improve the breastfeeding rate and reduce the occurrence of the complications, such as NEC, BPD, and ROP.
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Aleitamento Materno , Enterocolite Necrosante , Humanos , Lactente , Recém-Nascido , Recém-Nascido Prematuro , Unidades de Terapia Intensiva Neonatal , Método CanguruRESUMO
Objective: To investigate the clinical application of combined multiple artery-first approach to pancreatoduodenectomy. Methods: The clinical data of 53 patients who were diagnosed with peripancreatic head tumor at Department of Biliary-Pancreatic Surgery of Second Affiliated Hospital of Harbin Medical University between June 2013 and June 2015 was retrospectively analyzed.Pancreatic enhanced CT scan, magnetic resonance cholangiopancreatography, ultrasonography and tumor marker detection were applied for all the patients preoperatively.The 53 patients were operated by combined multiple artery-first approach(superior+ posterior approach, superior+ inferior approach, posterior+ inferior approach, superior+ posterior+ inferior approach) according to individualized therapeutic concept.And 42 patients underwent pancreatoduodenectomy, 9 patients underwent palliative operation and 2 patients just received exploratory operation. Results: Forty-two peripancreatic head tumor patients underwent pancreatoduodenectomy by applying combined multiple artery-first approach.The median operation time and intraoperative blood loss were (5.4±3.1)hours and (366±297)ml and the harvested lymph node and duration of hospital stay were 19±5 and (14.0±5.6)days.Nine patients underwent "total mesopancreas excision" and the rate of postoperative pancreatic fistula and R0 resection were 38.1% and 88.1%. Anomalous origin hepatic right artery was detected in one patients during the operation and no death occurred within 30 days postoperatively. Conclusion: According to the tumor location and patient's condition, individualistically applying combined multiple artery-first approach can reduce intraoperative blood loss, terminate unnecessary surgery, detect anomalous origin artery, make the tumor resection more radical and pancreatoduodenectomy more safety.
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Neoplasias Pancreáticas/cirurgia , Pancreaticoduodenectomia/métodos , Perda Sanguínea Cirúrgica , Humanos , Artéria Mesentérica Superior , Duração da Cirurgia , Pâncreas , Complicações Pós-Operatórias , Estudos Retrospectivos , Tomografia Computadorizada por Raios XRESUMO
Temozolomide (TMZ) resistance is an important cause of clinical treatment failure and poor prognosis in gliomas. Increasing evidence indicates that cancer-derived exosomes contribute to chemoresistance; however, the specific contribution of glioma-derived exosomes remains unclear. The aim of this study was to explore the role and underlying mechanisms of exosomal macrophage migration inhibitory factor (MIF) on TMZ resistance in gliomas. We first demonstrated that MIF was upregulated in the exosomes of TMZ-resistant cells, engendering the transfer of TMZ resistance to sensitive cells. Our results indicated that exosomal MIF conferred TMZ resistance to sensitive cells through the enhancement of cell proliferation and the repression of cell apoptosis upon TMZ exposure. MIF knockdown enhanced TMZ sensitivity in resistant glioma cells by upregulating Metalloproteinase Inhibitor 3 (TIMP3) and subsequently suppressing the PI3K/AKT signaling pathway. Additionally, exosomal MIF promoted tumor growth and TMZ resistance of glioma cells in vivo, while IOS-1 (MIF inhibitor) promotes glioma TMZ sensitive in vivo. Taken together, our study demonstrated that exosome-mediated transfer of MIF enhanced TMZ resistance in glioma through downregulating TIMP3 and further activating the PI3K/AKT signaling pathway, highlighting a prognostic biomarker and promising therapeutic target for TMZ treatment in gliomas.
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Prophylactic anti-D is a very safe and effective therapy for the suppression of anti-D immunization and thus prevention of haemolytic disease of the foetus and newborn. However, migration from countries with low health standards and substantial cuts in public health expenses have increased the incidence of anti-D immunization in many "developed" countries. Therefore, this forum focuses on prenatal monitoring standards and treatment strategies in pregnancies with anti-D alloimmunization. The following questions were addressed, and a response was obtained from 12 centres, mainly from Europe.
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Antígenos de Grupos Sanguíneos/imunologia , Isoanticorpos/administração & dosagem , Complicações Hematológicas na Gravidez/terapia , Isoimunização Rh/terapia , Sistema do Grupo Sanguíneo Rh-Hr/imunologia , Feminino , Sangue Fetal/imunologia , Hemoglobina Fetal/análise , Humanos , Isoanticorpos/sangue , Isoanticorpos/imunologia , Gravidez , Complicações Hematológicas na Gravidez/sangue , Complicações Hematológicas na Gravidez/imunologia , Complicações Hematológicas na Gravidez/prevenção & controle , Isoimunização Rh/imunologia , Isoimunização Rh/prevenção & controle , Imunoglobulina rho(D)RESUMO
Induction of liver allograft immunological tolerance was performed in rats by intramuscular injection of recombinant adeno-associated virus-human cytotoxic T-lymphocyte-associated antigen-4 immunoglobulin (rAAV-hCTLA4Ig). Dark Agouti and Lewis rats were liver allograft donors and recipients, respectively, in four groups: (A) syngeneic control, (B) blank control, (C) rAAV-enhanced green fluorescent protein negative control, (D) rAAV-hCTLA4Ig. Gene transfers occurred 6 weeks before transplantation. Group D had a significantly longer liver graft survival time (> 100 days) than groups B (11.9 +/- 1.3 days) and C (11.6 +/- 1.1 days). Groups B and C showed severe rejection responses and large amounts of CD4(+) and CD8(+) T-lymphocyte infiltration, while only a mild response and few T-lymphocytes were observed in group D. There were no significant differences in interleukin-2 and interferon-gamma levels in liver grafts between groups D and C, but there were significant decreases in granzyme B and lymphotoxin beta levels in group D compared with group C. It is concluded that immunological tolerance to liver allograft could be achieved by gene transfer of rAAV-hCTLA4Ig through intramuscular injection.
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Antígenos CD/genética , Antígenos CD/imunologia , Dependovirus/genética , Imunoglobulinas/imunologia , Transplante de Fígado/imunologia , Tolerância ao Transplante/imunologia , Animais , Antígenos CD/administração & dosagem , Antígenos CD/metabolismo , Antígeno CTLA-4 , Regulação da Expressão Gênica , Humanos , Injeções Intramusculares , Masculino , Músculos/metabolismo , RNA Mensageiro/genética , Ratos , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/imunologia , Taxa de SobrevidaRESUMO
The clinical features of the maternal syndrome of pre-eclampsia can be explained by generalised maternal endothelial cell dysfunction, which is a part of a more global maternal systemic inflammatory response. There is growing evidence that these effects are associated with the shedding of cellular debris, including syncytiotrophoblast microparticles (STBM), cell-free DNA and mRNA, from the surface of the placenta (syncytiotrophoblast) into the maternal circulation. The increased shedding of this debris seen in pre-eclampsia is believed to be caused by placental ischaemia, reperfusion and oxidative stress. This study was carried out to determine whether uterine contractions during labour and subsequent placental separation lead to an acute increase in the release of placental debris into the maternal circulation. To assess the effects of labour, samples were taken from 10 normal pregnant (NP) and 10 pre-eclamptic (PE) women at varied time points. Similarly to assess the effects of placental delivery, plasma samples were taken from 10 NP and 10 PE women undergoing elective caesarean section. There was a significant increase in the shedding of STBM in pre-eclampsia which was not seen in normal pregnancy and there was a small rise in STBM levels at placental separation in both normal pregnant and pre-eclamptic women undergoing caesarean section, but the differences were not significant. However, levels of placental cell-free corticotrophin releasing hormone mRNA were significantly increased in labour in both normal pregnancy and pre-eclampsia and were still high 24 h after delivery in the pre-eclamptic women. There was no significant increase in fetal or total DNA in labour, but the overall levels of total DNA (maternal and fetal) was increased in labour in pre-eclampsia compared to normal labour. The enhanced shedding of STBM and CRH mRNA in pre-eclampsia labour may have a role in cases of postpartum worsening of pre-eclampsia.
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Trabalho de Parto/fisiologia , Pré-Eclâmpsia/sangue , Pré-Eclâmpsia/patologia , Trofoblastos/patologia , Trofoblastos/fisiologia , Adulto , Cesárea , DNA/sangue , Feminino , Humanos , Tamanho da Partícula , Pré-Eclâmpsia/fisiopatologia , Gravidez , RNA Mensageiro/sangueRESUMO
AIMS: Cripto-1 may be capable of up-regulating signalling molecules associated with epithelial-to-mesenchymal transition (EMT), an important event characterized by loss of E-cadherin during malignant tumour progression and metastasis. The aim was to investigate the expression of Cripto-1 and E-cadherin in relation to clinicopathological features and patient prognosis of gastric cancer. METHODS AND RESULTS: The expression of Cripto-1 and E-cadherin was studied by immunohistochemistry in 118 gastric cancer cases. Up-regulated Cripto-1 (CR+) was found in 54% (64/118) of cases, whereas down-regulated E-cadherin (E-cad-) was found in 70% (83/118) of cases. Either CR+ or E-cad- was associated with lymph node metastasis, liver metastasis and late TNM stage (P < 0.05). Patients with either CR- or E-cad+ showed higher 5-year survival rates than those with CR+ or E-cad- (P = 0.0012 and P = 0.0017, respectively). When combined, evaluation of these two proteins, simultaneous CR+ and E-cad- (CR+/E-cad-) in cancer was strongly associated with the above three aggressive clinicopathological features (P < 0.001) and indicated the worst patient survival (P = 0.0001). Multivariate analysis revealed that CR+/E-cad- was an independent prognostic factor in gastric cancer. CONCLUSIONS: Combined analysis of Cripto-1 and E-cadherin has significant value in evaluating the metastatic potential of gastric cancer and predicting patient prognosis.
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Adenocarcinoma/metabolismo , Caderinas/metabolismo , Fator de Crescimento Epidérmico/metabolismo , Glicoproteínas de Membrana/metabolismo , Proteínas de Neoplasias/metabolismo , Neoplasias Gástricas/metabolismo , Adenocarcinoma/mortalidade , Adulto , Idoso , Idoso de 80 Anos ou mais , Progressão da Doença , Regulação para Baixo , Feminino , Proteínas Ligadas por GPI , Gastrectomia , Humanos , Técnicas Imunoenzimáticas , Peptídeos e Proteínas de Sinalização Intercelular , Linfonodos/patologia , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Prognóstico , Neoplasias Gástricas/mortalidade , Taxa de Sobrevida , Regulação para CimaRESUMO
OBJECTIVES: In order to assess the potential biomolecules for breast cancer, we analyzed in parallel the levels of cell-free glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and cell-free nucleosomes in serum samples from patients with benign and malignant breast tumors. The levels of cell-free DNA obtained by quantitative PCR were compared with those obtained by enzyme-linked immunosorbent assay (ELISA). METHODS: Twenty-three patients with benign breast tumors, 27 patients with breast cancer, and 32 age-matched healthy women were recruited. The amounts of serum nucleosomes were analyzed by ELISA and the levels of cell-free GAPDH were measured by real-time quantitative PCR. The correlation between nucleosome and cell-free GAPDH levels was examined using the Spearman rank test. RESULTS: The levels of cell-free GAPDH were significantly higher in the serum samples of patients with benign and malignant breast tumors than in those of the control group (median 37,966 GE/mL, range 3,802-130,104 versus 11,770 GE/mL, range 2,198-73,522, p=0.035 and median 40,698 GE/mL, range 3,644-192,482 versus 11,770 GE/mL range 2,198-73,522, p=0.001). The concentration of cell-free GAPDH correlated significantly with the quantities of nucleosomes in serum samples (r=0.451, p=0.000). There was, however, no significant difference between healthy individuals and women with benign breast tumors or breast cancer in terms of nucleosomes determined by ELISA. CONCLUSION: Our data suggest that the cell-free serum GAPDH DNA assayed by quantitative PCR is a better biomarker than nucleosomes assayed by ELISA in patients with breast tumors.
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Biomarcadores Tumorais/sangue , Neoplasias da Mama/diagnóstico , DNA/sangue , Ensaio de Imunoadsorção Enzimática/métodos , Nucleossomos , Reação em Cadeia da Polimerase/métodos , Neoplasias da Mama/sangue , Feminino , Gliceraldeído-3-Fosfato Desidrogenases/genética , HumanosRESUMO
PURPOSES OF THE STUDY: We analyzed circulating cell-free DNA in the serum of patients with benign and malignant breast disease and in healthy individuals to determine its diagnostic value. BASIC PROCEDURES: Serum samples were obtained from 50 healthy individuals, 33 patients with malignant breast disease and 32 patients with benign breast disease. Circulatory DNA was extracted from serum samples. Cell-free DNA was quantified by real-time quantitative PCR for the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene. Tissue samples from patients with malignant and benign breast lesions were histopathologically examined. MAIN FINDINGS: The mean levels of circulating cell-free DNA in serum samples were 41,149 genome equivalents (GE)/mL in patients with malignant disease, 30,826 GE/mL in patients with benign disease, and 13,267 GE/mL in healthy individuals. Healthy individuals had significantly lower levels of cell-free DNA than patients with malignant or benign breast disease (p=0.001, p=0.031). No significant difference was observed between malignant and benign disease. There was a correlation between cell-free DNA levels and tumor size but not with other tumor characteristics. PRINCIPAL CONCLUSION: Our results suggest that levels of circulating cell-free DNA in serum could have diagnostic value to discriminate between healthy individuals and patients with breast lesions but not between patients with malignant and benign breast lesions.
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Doenças Mamárias/sangue , Neoplasias da Mama/sangue , DNA de Neoplasias/sangue , DNA/sangue , Adulto , Idoso , Neoplasias da Mama/classificação , DNA/genética , DNA/isolamento & purificação , DNA de Neoplasias/genética , DNA de Neoplasias/isolamento & purificação , Genoma Humano , Gliceraldeído-3-Fosfato Desidrogenases/genética , Humanos , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Valores de ReferênciaRESUMO
Reduced intracellular accumulation of drugs mediated by efflux pump is one of the most critical mechanisms governing fluconazole (FLC) resistance in Candida albicans (C. albicans). Besides, mitochondrial aerobic respiration plays a major role in C. albicans metabolism. However, it is unclear whether mitochondrial aerobic respiration is involved with efflux-mediated resistance of C. albicans to azole. We measured key parameters of energy conversion, including the activity of respiratory chain complexes I, III and V (CI, CIII and CV), and reactive oxygen species (ROS) in two C. albicans strains (FLC-susceptible strain CA-1S and FLC-resistant strain CA-16R) obtained from a single parental source. Additionally, we quantified intracellular ATP levels and mitochondrial membrane potential (ΔΨm), which has critical effect on energy transport. Our analyses revealed a higher ATP level and ΔΨm in CA-16R compared with CA-1S (P<0.05), and a higher ATP level and ΔΨm in Sc5314S (FLC-susceptible strain) compared with Sc5314R (FLC-resistant strain). CI and CV activity increased in CA-16R, activity of CI, CIII and CV increased in Sc5314R. Additionally, ROS decreased in CA-16R and Sc5314R compared with their respective susceptible counterparts. Our data suggest that mitochondrial aerobic respiratory metabolism might be directly associated with the efflux-mediated resistance of C. albicans to azole. C. albicans strains might enhance the activity of efflux pumps and therefore decrease sensitivity to FLC through alteration of mitochondrial aerobic respiratory metabolism, by increased ATP production and decreased ROS generation.
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Candida albicans/fisiologia , Respiração Celular/efeitos dos fármacos , Farmacorresistência Fúngica/fisiologia , Fluconazol/farmacologia , Transportadores de Cassetes de Ligação de ATP/metabolismo , Transportadores de Cassetes de Ligação de ATP/fisiologia , Aerobiose/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Candida albicans/patogenicidade , Respiração Celular/fisiologia , Metabolismo Energético/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Mitocôndrias/fisiologia , Espécies Reativas de Oxigênio/metabolismoRESUMO
Electron energy-loss magnetic chiral dichroism (EMCD) spectroscopy, which is similar to the well-established X-ray magnetic circular dichroism spectroscopy (XMCD), can determine the quantitative magnetic parameters of materials with high spatial resolution. One of the major obstacles in quantitative analysis using the EMCD technique is the relatively poor signal-to-noise ratio (SNR), compared to XMCD. Here, in the example of a double perovskite Sr2FeMoO6, we predicted the optimal dynamical diffraction conditions such as sample thickness, crystallographic orientation and detection aperture position by theoretical simulations. By using the optimized conditions, we showed that the SNR of experimental EMCD spectra can be significantly improved and the error of quantitative magnetic parameter determined by EMCD technique can be remarkably lowered. Our results demonstrate that, with enhanced SNR, the EMCD technique can be a unique tool to understand the structure-property relationship of magnetic materials particularly in the high-density magnetic recording and spintronic devices by quantitatively determining magnetic structure and properties at the nanometer scale.
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The independent control of two magnetic electrodes and spin-coherent transport in magnetic tunnel junctions are strictly required for tunneling magnetoresistance, while junctions with only one ferromagnetic electrode exhibit tunneling anisotropic magnetoresistance dependent on the anisotropic density of states with no room temperature performance so far. Here, we report an alternative approach to obtaining tunneling anisotropic magnetoresistance in α'-FeRh-based junctions driven by the magnetic phase transition of α'-FeRh and resultantly large variation of the density of states in the vicinity of MgO tunneling barrier, referred to as phase transition tunneling anisotropic magnetoresistance. The junctions with only one α'-FeRh magnetic electrode show a magnetoresistance ratio up to 20% at room temperature. Both the polarity and magnitude of the phase transition tunneling anisotropic magnetoresistance can be modulated by interfacial engineering at the α'-FeRh/MgO interface. Besides the fundamental significance, our finding might add a different dimension to magnetic random access memory and antiferromagnet spintronics.Tunneling anisotropic magnetoresistance is promising for next generation memory devices but limited by the low efficiency and functioning temperature. Here the authors achieved 20% tunneling anisotropic magnetoresistance at room temperature in magnetic tunnel junctions with one α'-FeRh magnetic electrode.
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This study evaluated the specificity and sensitivity of the reverse transcriptase/polymerase chain reaction (RT-PCR) for carcinoembryonic antigen (CEA) for identification of breast tumour cells in bone marrow and in peripheral blood. Using one primer set from the A2/B3 domains of CEA with five to seven mismatches to other CEA-family members allowed reproducible detection of 1 colon tumour cell and 10 breast tumour cells in 10(7) mononuclear cells. Bone marrow samples from 181 patients with breast cancer were analysed by CEA-RT-PCR; 50 of these samples were analysed in parallel by routine immunocytochemistry. CEA-mRNA-positive bone marrow cells were found in 27.6% of the patients (50/181) with breast cancer. Five immunocytochemistry-positive samples were negative when analysed by CEA-RT-PCR. Limiting factors in the detection of micrometastatic breast tumour cells by CEA-RT-PCR are the heterogeneity of the tumour cells and the deficient expression of CEA in some of these cells. However, CEA-RT-PCR using the specific primer could detect 1 colon tumour cell in 1 x 10(7) normal peripheral blood mononuclear cells.
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Medula Óssea/química , Neoplasias da Mama/metabolismo , Antígeno Carcinoembrionário/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Medula Óssea/metabolismo , Neoplasias da Mama/sangue , Neoplasias da Mama/genética , Antígeno Carcinoembrionário/genética , Estudos de Avaliação como Assunto , Feminino , Humanos , Imuno-Histoquímica , RNA Mensageiro/genética , Sensibilidade e Especificidade , Células Tumorais CultivadasRESUMO
PURPOSE: The work aimed to evaluate the sensitivity and specificity of the cytokeratin (CK) 19 reverse transcriptase/polymerase chain reaction (RT-PCR) for the detection of occult breast cancer in bone marrow and leukapheresis products. MATERIALS AND METHODS: Peripheral blood and bone marrow samples, obtained from 96 and 8 healthy donors respectively, served as negative controls. A total of 115 bone marrow samples and 29 leukapheresis samples from routine patients with breast cancer were analysed by CK19 RT-PCR. The PCR results were compared with those from routine immunocytology for CK8, 18, 19. RESULTS: The CK19 RT-PCR technique with primer pairs from Datta et al. (J Clin Oncol 12: 475-482, 1994), using an annealing temperature of 72 degrees C, allowed the detection of one tumour cell in 10(7) mononuclear cells. None of the control samples (96 peripheral blood and 8 bone marrow) that were positive for beta2-microglobulin by RT-PCR showed a signal for CK19. However, expression of CK19 mRNA was observed in 40.87% (70/115) of bone marrow and in 24.13% (7/29) of leukapheresis samples of patients with breast cancer. Standard immunocytology and PCR were combined for the detection of tumour cells. Five of the 65 bone marrow samples were found to be positive by CK19 RT-PCR, but were negative with the immunocytology method. CONCLUSION: RT-PCR using CK19-specific primers and optimal experimental conditions is a reliable and specific method for the detection of micrometastatic breast cancer cells.
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Neoplasias da Medula Óssea/química , Neoplasias da Medula Óssea/secundário , Neoplasias da Mama/química , Neoplasias da Mama/patologia , Queratinas/análise , Leucaférese , Primers do DNA , Feminino , Humanos , Queratinas/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e EspecificidadeRESUMO
We report a highly sensitive method to detect rare human breast cancer cells, which combines an immunomagnetic separation (IMS) using antibody BM2 against MUC-1 with cytokeratin-19 (CK19) and the reverse transcriptase/polymerase chain reaction (RT-PCR). The IMS-RT-PCR technique allows the detection of 1 tumor cell/10(7)-10(8) mononuclear cells. This is at least ten times more sensitive than CK19 RT-PCR alone, or immunocytochemistry. All 117 peripheral blood and 8 bone marrow samples obtained from healthy donors as negative controls were positive for beta2-microglobulin by RT-PCR but negative for CK19 by IMS-RT-PCR or RT-PCR alone. Out of 26 bone marrow samples from breast cancer patients, 18 had CK19 transcripts detectable by IMS-RT-PCR. In contrast, only 14 and 13 samples from the 26 were found to be positive by RT-PCR alone or by routine immunocytochemical staining. In conclusion, IMS-RT-PCR for CK19 is a highly sensitive and specific method for detecting very low numbers of micrometastatic breast cancer cells in bone marrow amidst an excess of nonmalignant cells. For the early diagnosis of disseminating disease, this assay is more efficient than RT-PCR alone and routine immunocytochemistry.
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Neoplasias da Medula Óssea/secundário , Neoplasias da Mama/patologia , Separação Imunomagnética/métodos , Queratinas/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Medula Óssea/metabolismo , Medula Óssea/patologia , Neoplasias da Medula Óssea/metabolismo , Neoplasias da Medula Óssea/patologia , Neoplasias da Mama/metabolismo , Separação Celular , Feminino , Humanos , Técnicas In Vitro , Queratinas/genética , Células Neoplásicas Circulantes , Sensibilidade e EspecificidadeRESUMO
Elevations in fetal cell traffic as well as increased release of cell-free fetal DNA into the maternal periphery have previously been shown to occur in pregnancies affected by preeclampsia. Our own investigations have shown that manifestation of preeclampsia is associated with an increased accumulation of circulatory fetal DNA as well as cell-free maternal DNA in maternal plasma. We further established that the increments in these two molecular genetic analytes corresponded to the severity of the disease and to each other. This latter phenomenon was evident in preeclamptic pregnancies, but not in normal ones. As we had recently performed a prospective study to investigate fetal cell traffic prior to onset of preeclamptic symptoms, we examined the levels of cell-free fetal and maternal DNA in these samples. This analysis indicated that circulatory fetal DNA concentrations were significantly elevated prior to onset of the disease symptoms. No similar feature was observed for cell-free maternal DNA levels.
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DNA/sangue , Troca Materno-Fetal , Pré-Eclâmpsia/sangue , Gravidez de Alto Risco , Feminino , Humanos , GravidezRESUMO
Elevations in the concentration of cell-free fetal DNA in maternal plasma have recently been determined in various pregnancy-related disorders, including preeclampsia, preterm labor, and polyhydramnios. In addition, almost 2-fold increments in cell-free fetal DNA levels have been recorded in pregnancies with certain aneuploid fetuses, in particular trisomy 21. These findings have led to the speculation that quantitative assessment of circulatory fetal DNA may be useful in the noninvasive prenatal diagnosis of certain fetal genetic constellations or pregnancy-related disorders. A premise for any quantitative analysis is that the quantity of the analyte being assayed does not vary greatly over time. As this aspect has not been examined for circulatory DNA levels, we examined these in normal healthy individuals as well as in pregnant women. Our data indicate that severalfold alterations in circulatory DNA amounts do occur over short periods of time. Of particular note is that we observed almost 2-fold variations in free fetal DNA levels over a period of 3 days, which are in a similar range to the elevations noted in aneuploid pregnancies. Our results, therefore, imply that caution should be used when using small increments in circulatory fetal DNA concentrations for potential diagnostic applications.