RESUMO
Objective: To evaluate the effect of D-dimer on the prognosis of patients with aneurysmal subarachnoid hemorrhage (aSAH). Methods: A total of 1 658 patients who were first diagnosed with aSAH in West China Hospital of Sichuan University from December 2013 to June 2019 were retrospectively analyzed. All patients were divided into four groups according to the median and quartiles of D-dimer level, including 415 cases, 414 cases, 414 cases, and 415 cases in groups Q1, Q2, Q3, and Q4, respectively. Groups Q2, Q3, Q4, and group Q1 were matched by propensity score matching (PSM), and the correlation between D-dimer and each outcome was analyzed by logistic regression. Since there is no general clinical classification standard for D-dimer, this study attempted to reclassify patients into groups q1 (<0.55 mg/L, 94 cases), q2 (0.55-1.65 mg/L, 435 cases), q3 (1.65-5.50 mg/L, 650 cases) and q4 (>5.50 mg/L, 303 cases) based on 1, 3, 5, 10 times of the upper limit of the current clinical reference value. Results: The age of 1 658 aSAH patients were (57±12) years, including 1 068 males and 590 females. After PSM based on the median and quartiles of D-dimer level, there were 318 cases, 318 cases, 251 cases, and 229 cases in groups Q1, Q2, Q3, and Q4, respectively. Compared with group Q1 (<1.23 mg/L), the risk of in-hospital infection (OR=2.14, 95%CI: 1.47-3.11, P<0.001), pneumonia (OR=2.22, 95%CI: 1.51-3.28, P<0.001), urinary tract infection (OR=1.75, 95%CI: 1.12-2.75, P=0.014) and intracranial rebleeding (OR=3.59, 95%CI: 1.30-9.91, P=0.013) group Q4 (>4.95 mg/L) was higher. Likewise, the risk of adverse outcomes in group Q4 was also higher than that in group Q1, including unfavorable outcome at discharge (OR=2.12, 95%CI: 1.43-3.14, P<0.001), mortality during hospitalization (OR=3.03, 95%CI: 1.26-7.33, P=0.014), mortality within 90 days (OR=2.33, 95%CI:1.29-4.22, P=0.005), mortality within 180 days (OR=1.92, 95%CI: 1.12-3.29, P=0.018), mortality within 1 year (OR=2.07, 95%CI:1.23-3.47, P=0.006) and mortality during the longest follow-up period (OR=1.97, 95%CI:1.26-3.09, P=0.003). After secondary grouping and PSM based on current clinical reference values, there were 90 cases, 87 cases, 90 cases, and 43 cases, respectively in groups q1, q2, q3 and q4. The risk of nosocomial infection (OR=2.26, 95%CI: 1.14-4.45, P=0.019), blood-borne infection (OR=8.86, 95%CI:1.08-72.78, P=0.042), poor prognosis at discharge (OR=4.92, 95%CI: 2.18-11.07, P<0.001), death within 180 days (OR=3.39, 95%CI: 1.04-11.08, P=0.043), death within 1 year (OR=3.23, 95%CI: 1.10-9.49, P=0.033), and death within the longest follow-up period (OR=3.28, 95%CI: 1.34-8.01, P=0.009) was still higher in group q4 than that in group q1. Conclusion: aSAH patients with high D-dimer level have a higher risk of complications and mortality during hospitalization and worse clinical prognosis.
Assuntos
Hemorragia Subaracnóidea , Idoso , Feminino , Produtos de Degradação da Fibrina e do Fibrinogênio , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Pontuação de Propensão , Estudos RetrospectivosRESUMO
Objective: To construct zebrafish models for the screening of intracranial hemorrhage (ICH) associated genes. Methods: ICH zebrafish models were constructed through morpholino oligonucleotides (MOs) technique and microinjection technique, and multiple verification was performed from macro and micro perspectives. First, the normal wild-type AB strain zebrafish injected with control MO was used as the control group, and AB zebrafish embryos microinjected with MOs of genes related to development of neural crest-derived cells (NCDCs) were used as the study group, such as col8a1 MO, tfap2α MO, msx1a MO, msx2 MO, and dkk1a MO. Preliminary verification of the model was conducted under a white-light optical microscope. Then, the model was verified by Tg (flk1: gfp; gata1: dsRed) double transgenic zebrafish, with vascular endothelial cells labeled by green fluorescent protein (GFP) and red blood cell labeled by fluorescent protein (dsRed), and thus the location of cerebral hemorrhage can be observed more clearly. Specifically, zebrafish embryos were microinjected with Control MO as the control group and those microinjected with col8a1 MO as the study group. Then the embryos were cultured until 48 hours post-fertilization to observe the leakage of red blood cells under the confocal laser scanning microscope. Finally, Tg (flk1: gfp) transgenic zebrafish was used to verify the model based on the blood-brain barrier (BBB). Through the leakage of dextran-rhodamine and DAPI dyes, the destruction of BBB and the occurrence of cerebral hemorrhage in zebrafish were further clarified, and quantitative statistics were carried out to verify the relationship between NCDCs development related genes and cerebral hemorrhage phenotype, which proved that the modeling was effective. Results: The zebrafish with col8a1, tfap2α, and msx1 mutations in the study group had apparent ICH compared with wildtype zebrafish, and the prevalence of ICH was 18.18% (52/286), 23.04% (62/251), and 35.94% (23/64), respectively. While, the zebrafish with msx2 and dkk1a mutations rarely had ICH, with the ICH prevalence of 1.03% (1/97) and 1.15% (1/87), respectively. The prevalence of red blood cells leakage in Tg (flk1:gfp; gata1:dsred) double transgenic zebrafish injected with Control Mo and col8a1 Mo was 0.37% (1/273) and 18.18% (52/286) (P<0.001). The number of DAPI positive nuclei of Tg (flk1: gfp) transgenic zebrafish injected with Control Mo and col8a1 Mo was 10.05±5.27 and 60.35±3.96 (P<0.001), and the fluorescent intensity of midbrain parenchymal induced by dextran-rhodamin leakage was 2.54±4.70 and 5.13±3.52 (P<0.001). Conclusion: This study successfully constructs the ICH zebrafish models, and ICH-related genes are screened out, such as col8a1, tfap2α, msx1, and so on.
Assuntos
Células Endoteliais , Peixe-Zebra , Animais , Animais Geneticamente Modificados , Hemorragia Cerebral , Dextranos , Proteínas de Fluorescência Verde , Peixe-Zebra/genéticaRESUMO
Hepatocellular carcinoma (HCC) is a major disease with a high degree of malignancy, and poor prognosis, which seriously endangers human health. Chronic viral hepatitis (HBV, HCV)-cirrhosis-liver cancer patterns of pathogenesis has been widely accepted. However, the relationship between non-infectious liver disease and HCC is not completely clear; thereby how various non-infectious liver diseases develop through precancerous lesions has attracted widespread attention to HCC. A full understanding of these precancerous lesions is likely to provide new ideas and strategies for the prevention and treatment of non-infectious liver disease-related HCC.
Assuntos
Carcinoma Hepatocelular , Hepatite Crônica , Cirrose Hepática , Neoplasias Hepáticas , Lesões Pré-Cancerosas/diagnóstico , HumanosRESUMO
OBJECTIVE: To determine whether chloroquine (CQ), an often used inhibitor of late autophagy and autophagosome/lyosome fusion, can inhibit proliferation of renal carcinoma cells and investigate its effect on sunitinib (ST)-induced apoptosis. METHODS: Renal carcinoma cell line 786 O and ACHN had been used as cellular model and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay was carried out to detect the cell viability in response to CQ or ST treatment. Both transmission electron microscope and immunoblotting had been employed to observe apoptotic and autophagic process. To examine the involvement of autophagy in ST-dependent apoptosis, autophagy had been inhibited either chemically or genetically via utilizing autophagy inhibitor or specific small interference RNA (siRNA) targeted to either Ulk1 (unc-51-like kinase 1) or LC3 (microtubule associated protein 1 light chain 3 fusion protein), two essential autophagic proteins. RESULTS: Both ST and CQ induced cell viability loss, indicating that either of them could inhibit renal cancer cell proliferation. Clone formation experiments confirmed the aforementioned results. Furthermore, the combined ST with CQ synergistically promoted the loss of cell viability. By transmission electron microscopy and immunoblotting, we found that the ST induced both autophagy and caspase-dependent apoptosis. While 3-MA, an early autophagy inhibitor, reduced the ST-induced cleavage of poly (ADP-ribose) polymerase-1 (PARP-1), a substrate of caspase 3/7 and often used marker of caspase-dependent apoptosis, CQ promoted the ST-dependent PARP-1 cleavage, indicating that the early and late autophagy functioned differentially on the ST-activated apoptotic process. Moreover, the knock down of either Ulk1 or LC3 decreased the ST-caused apoptosis.Interestingly, we observed that rapamycin, a specific inhibitor of mTOR (mammalian target of rapamycin) and an inducer of autophagy, also showed to inhibit cell viability and increased the cleavage of PARP-1 in the ST-treated cells, suggesting that autophagy was likely to play a dual role in the regulation of the ST-induced apoptosis. CONCLUSION: ST activates both apoptotic and autophagic process in renal carcinoma cells. Although autophagy precedes the ST-induced apoptosis, however, early and late autophagy functions differentially on the apoptotic process induced by this compound. Additionally, ST can coordinate with the inducer of autophagy to inhibit the cell proliferation. Further research in this direction will let us illuminate to utilize CQ as a potential drug in the treatment of renal carcinoma.
Assuntos
Antineoplásicos , Antirreumáticos , Apoptose , Cloroquina , Neoplasias Renais , Sunitinibe , Animais , Antineoplásicos/farmacologia , Antirreumáticos/farmacologia , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Caspases , Linhagem Celular Tumoral , Cloroquina/farmacologia , Neoplasias Renais/tratamento farmacológico , Sunitinibe/farmacologiaRESUMO
Objective: To investigate the efficiency of FibroScan(FS)and FibroTouch(FT)in liver stiffness measurement(LSM)and fat quantification through a comparative analysis. Methods: The outpatients or hospitalized patients who underwent LSM and fat quantification using FS and FT were enrolled. The differences in success rate and detecting parameters between FS and FT were analyzed, as well as the correlation between FS and FT values. The t-test was used for comparison of normally distributed continuous data between groups, and a one-way analysis of variance or the Kruskal-Wallis test was used for comparison between multiple groups. The Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups. Results: A total of 1621 patients were enrolled. The success rates of FT and FS were 100% and 94.96%, respectively, and the success rate of FS was influenced by sex, age, body mass index, and biochemical markers of liver function. FT has a significantly shorter duration of single detection and a significantly lower number of times of single detection than FS(duration of single detection: 190.21±38.78 s vs 220.89±68.36 s, P < 0.01; number of single detection times: 10.31±1.32 vs 11.81±3.76, P < 0.01), as well as a significantly lower ratio of interquartile range to median of fat quantification in the same patient(5.39%±4.81% vs 17.18%±14.07%, P < 0.01). The LSM and fat quantification of FS were significantly correlated with those of FT(r = 0.645 and 0.620, both Based on the duration and number of times of single detection, success rate, and stability of fat quantification, FT seems to have a better detection efficiency than FS. The detection values of FT and FS can be calculated with regression equations < 0.01). The equations of linear regression were LSM(FT)= 4.435+0.477×LSM(FS); CAP(FT)= 134.71+0.456×CAP(FS). Conclusion: Based on the duration and number of times of single detection, success rate, and stability of fat quantification, FT seems to have a better detection efficiency than FS. The detection values of FT and FS can be calculated with regression equations.
Assuntos
Técnicas de Imagem por Elasticidade/métodos , Fígado Gorduroso/sangue , Fígado Gorduroso/diagnóstico por imagem , Fígado/diagnóstico por imagem , Adulto , Biomarcadores/sangue , Fígado Gorduroso/patologia , Feminino , Humanos , Fígado/patologia , Cirrose Hepática/diagnóstico , Masculino , Pessoa de Meia-Idade , Hepatopatia Gordurosa não Alcoólica , Valor Preditivo dos Testes , Sensibilidade e Especificidade , Fatores de TempoRESUMO
The incidence of papillary thyroid carcinoma is growing rapidly. Especially in young patients, the patients have high demand for better cosmetic effect and less psychological trauma. The application of laparoscope technique in papillary thyroid carcinoma surgery should meet the needs of patients. The development of needle assisted technique make it easy to access the neck potential fascial spaces, and provide the basic instrument for laparoscope modified neck dissection. The application of needle assisted techniques mainly include suture suspension and V type needle retractor, MiniLap, 3 mm laparoscopic apparatus. Indications of this procedure is very important, and it is necessary to make full use of the "critical points" , "mark lines" and "fascia layers" to precise positioning in the operation. Furthermore, the levels of dissection must be determined based on the pattern of cervical lymph node metastasis in differentiated thyroid carcinoma, that will make the surgical procedures more safety.
Assuntos
Carcinoma/cirurgia , Esvaziamento Cervical/métodos , Neoplasias da Glândula Tireoide/cirurgia , Tireoidectomia , Mama , Carcinoma Papilar , Humanos , Laparoscópios , Linfonodos , Metástase Linfática , Pescoço , Agulhas , Câncer Papilífero da TireoideRESUMO
Although many studies have been carried out on monoclonal gammopathy of unknown significances (MGUS), smoldering multiple myeloma (SMM), and multiple myeloma (MM), their classification and underlying pathogenesis are far from elucidated. To discover the relationships among MGUS, SMM, and MM at the transcriptome level, differentially expressed genes in MGUS, SMM, and MM were identified by the rank product method, and then co-expression networks were constructed by integrating the data. Finally, a pathway-network was constructed based on Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis, and the relationships between the pathways were identified. The results indicated that there were 55, 78, and 138 pathways involved in the myeloma tumor developmental stages of MGUS, SMM, and MM, respectively. The biological processes identified therein were found to have a close relationship with the immune system. Processes and pathways related to the abnormal activity of DNA and RNA were also present in SMM and MM. Six common pathways were found in the whole process of myeloma tumor development. Nine pathways were shown to participate in the progression of MGUS to SMM, and prostate cancer was the sole pathway that was involved only in MGUS and MM. Pathway-network analysis might provide a new indicator for the developmental stage diagnosis of myeloma tumors.
Assuntos
Redes Reguladoras de Genes , Redes e Vias Metabólicas , Mieloma Múltiplo/genética , Mieloma Múltiplo/metabolismo , Transdução de Sinais , Biologia Computacional , Conjuntos de Dados como Assunto , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Anotação de Sequência Molecular , Gamopatia Monoclonal de Significância Indeterminada/genética , Gamopatia Monoclonal de Significância Indeterminada/metabolismo , Gamopatia Monoclonal de Significância Indeterminada/patologia , Mieloma Múltiplo/patologia , Paraproteinemias/genética , Paraproteinemias/metabolismo , Paraproteinemias/patologiaRESUMO
Chinese herbal medicine Jinlianqingre Effervescent Tablets (JET) are the recommended control measure for uncomplicated hand, foot, and mouth disease (HFMD) by the Ministry of Health of China. However, high-quality evidence to support this recommendation is limited. A total of 288 patients ranging in age from 1 to 13 years were randomly assigned to JET in combination with conventional therapy (mainly including the reduction of temperature by applying physical cooling paste or warm bathing), or conventional therapy with placebo group for 7 days. The objective was to test the hypothesis that JET combination therapy is more effective than conventional therapy for uncomplicated HFMD. A randomized, double-blind, placebo-controlled trial was designed. Our study showed that, compared with conventional therapy, the median time to fever resolution was significantly shorter in the JET combination therapy (8 vs. 80 h; p < 0.0001); the risk of fever resolution increased in the JET combination therapy [hazard ratio, 19.8; 95% confidence interval (CI), 12.8 to 30.7]; the median healing time of rash or oral ulcer was significantly shorter in the JET combination therapy (14 vs. 74 h; p < 0.0001); and the median symptom score for skin or oral mucosa lesions improved more rapidly in the JET combination therapy during the follow-up period. The median duration of hospital stay was 6 days in the JET combination therapy and 7 days in the conventional therapy (p < 0.0001). No significant adverse events and complications were found in both groups. The addition of JET to conventional therapy reduced fever clearance time, healing time of skin or oral mucosa lesions, and duration of hospital stay in children with uncomplicated HFMD.
Assuntos
Doença de Mão, Pé e Boca/tratamento farmacológico , Adolescente , Criança , Pré-Escolar , Terapia Combinada , Método Duplo-Cego , Feminino , Humanos , Lactente , Tempo de Internação , Masculino , Resultado do TratamentoRESUMO
OBJECTIVE: We studied the efficacy of 25-microg misoprostol pessaries as either single or double dose compared with a 3-mg dinoprostone pessary for cervical priming. DESIGN AND SETTING: A randomised controlled trial in Singapore. POPULATION: One hundred and seventy-one women with term pregnancies and modified Bishop scores (mBS) < or =6 from 2003 to 2004. METHOD: Patients were randomised to single misoprostol dose, double misoprostol dose or the current dinoprostone regimen. MAIN OUTCOME MEASURES: Primary outcome was number of women who achieved favourable mBS >6 or active labour by day 2. Secondary outcomes were time interval from insertion to delivery, cardiotocographic abnormalities, delivery and neonatal outcome. RESULTS: More women in the misoprostol double-dose group (96.6%) and dinoprostone group (93%) achieved the primary outcome compared with the single-dose group (77.8%) (P = 0.003 and P = 0.03, respectively). There was no difference in secondary outcomes. More multiparous women achieve primary outcome compared with nulliparous women (odds ratio 0.21, 95% confidence interval 0.06-0.77). CONCLUSION: Double-dose misoprostol 25 microg is as effective as dinoprostone 3 mg inserts for cervical priming; both are more efficacious than a single-dose misoprostol pessary. Parity prognosticates the success of induction.
Assuntos
Maturidade Cervical/efeitos dos fármacos , Dinoprostona/administração & dosagem , Misoprostol/administração & dosagem , Ocitócicos/administração & dosagem , Administração Intravaginal , Adulto , Índice de Apgar , Dinoprostona/efeitos adversos , Quimioterapia Combinada , Feminino , Humanos , Recém-Nascido , Trabalho de Parto Induzido/métodos , Misoprostol/efeitos adversos , Análise Multivariada , Ocitócicos/efeitos adversos , Paridade/fisiologia , Pessários , Gravidez , Resultado da Gravidez , Análise de RegressãoRESUMO
Objective: To investigate the incidence and pathogen distribution of ventilator-associated pneumonia (VAP) among preterm infants admitted to level â ¢ neonatal intensive care units (NICU) in China. Method: A prospective study was conducted in 25 level â ¢ NICU, enrolling all preterm infants <34 weeks gestational age admitted to the participating NICU within the first 7 days of life from May 2015 to April 2016. Chi-square test, t test and Mann-Whitney U test were used for statistical analysis. Result: A total of 7 918 patients were enrolled, within whom 4 623(58.4%) were males. The birth weight was (1 639±415) g and the gestational age was (31.4±2.0) weeks; 4 654(58.8%) infants required non-invasive mechanical ventilation and 2 154(27.2%) required intubation. Of all the mechanically ventilated patients, VAP occurred in 95 patients. The overall VAP rate was 7.0 episodes per 1 000 ventilator days, varying from 0 to 34.4 episodes per 1 000 ventilator days in different centers. The incidence of VAP was 9.6 and 6.0 per 1 000 ventilator days in children's hospitals and maternity-infant hospitals respectively, without significant differences (t=1.002, P=0.327). Gram-negative bacilli (76 strains, 91.6%) were the primary VAP microorganisms, mainly Acinetobacter baumannii (24 strains, 28.9%), Klebsiella pneumonia (23 strains, 27.7%), and Pseudomonas aeruginosa (10 strains, 12.0%). Conclusion: The incidence of VAP in China is similar to that in developed counties, with substantial variability in different NICU settings. More efforts are needed to monitor and evaluate the preventable factors associated with VAP and conduct interventions that could effectively reduce the occurrence of VAP.
Assuntos
Idade Gestacional , Recém-Nascido Prematuro , Pneumonia Associada à Ventilação Mecânica , Peso ao Nascer , China , Feminino , Bactérias Gram-Negativas , Hospitais Pediátricos , Humanos , Incidência , Recém-Nascido , Unidades de Terapia Intensiva Neonatal , Klebsiella pneumoniae , Masculino , Estudos Prospectivos , Ventiladores MecânicosRESUMO
A genetically engineered, nonneurotropic herpes simplex virus (R7020) with a proven safety profile in both animals and humans was found effective in the treatment of large xenotransplanted tumors arising from a radiation- and chemotherapy-resistant human epidermoid carcinoma and a hormone-refractory prostate adenocarcinoma. R7020 replicated to high titer and caused rapid regression of the human tumor xenografts. Tumor destruction was accelerated in animals given both R7020 and fractionated ionizing radiation. Tumors arising from cells surviving one treatment with R7020 were fully susceptible to a second dose of virus. We conclude R7020 is an effective antitumor agent for non-central nervous system tumor xenografts with an excellent safety profile.
Assuntos
Adenocarcinoma/terapia , Carcinoma de Células Escamosas/terapia , Neoplasias da Próstata/terapia , Simplexvirus/fisiologia , Adenocarcinoma/genética , Adenocarcinoma/radioterapia , Animais , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/radioterapia , Fracionamento da Dose de Radiação , Resistencia a Medicamentos Antineoplásicos , Regulação Viral da Expressão Gênica/efeitos da radiação , Genes p53 , Engenharia Genética , Humanos , Injeções Intralesionais , Masculino , Camundongos , Camundongos Nus , Neoplasias da Próstata/genética , Neoplasias da Próstata/radioterapia , Tolerância a Radiação , Simplexvirus/genética , Transplante Heterólogo , Replicação ViralRESUMO
OBJECTIVE: The objective of this paper is to identify pathway-related modules which are defined as in high-grade osteosarcoma based on topological centralities analysis of networks. MATERIALS AND METHODS: Co-expression network was constructed by weighted gene co-expression network analysis (WGCNA) based on differentially expressed genes (DEGs). Pathway enrichment analysis was conducted by Kyoto Encyclopedia of Genes and Genomes (KEGG) database to detect pathway enriched genes. Pathway-related modules of pathway enriched genes were mined from the co-expression network. Then topological centralities (degree, closeness, stress and betweenness centrality) analyses for co-expression network and sub-networks were performed to explore hub genes. Validation of hub genes was carried out utilizing reverse transcription-polymerase chain reaction (RT-PCR) assays. RESULTS: There were 129 nodes and 1229 edges in co-expression network. We obtained a total of 16 hub genes and 11 pathway-related modules. Module 17 (Bladder cancer module) was the most significant module, which comprising 9 of 16 hub genes and 6 pathway enriched genes, taking intersection elements (CAV1 and CCND1). RT-PCR results showed that both of CAV1 and CCND1 in high-grade osteosarcoma were significantly differentially expressed compared with normal controls. CONCLUSIONS: This work may contribute to understanding the molecular pathogenesis and provide potential biomarkers for detections and effective therapies of high-grade osteosarcoma.
Assuntos
Perfilação da Expressão Gênica , Redes Reguladoras de Genes , Osteossarcoma/metabolismo , Biomarcadores , Neoplasias Ósseas , HumanosRESUMO
The nucleus of a blastula cell from Tilapia (Oreochromis nilotica, family Cichlidae, order Perciformes) was transplanted into an enucleated egg of Loach (Paramisgurnus dabryanus, family Cobitidae, order Cypriniformes). From among 3747 nucleo-cytoplasmic hybrid (NCH) eggs two NCH larval fish (0.05%) were obtained; one died on the 6th day and the other died on the 12th day after the operation. Morphological examinations showed that both NCH larval fish had developed normally with an opened mouth except they could not take food after complete utilization of their egg yolk on the 5th day of development. The possible mechanisms for obtaining such inter-order NCH larval fish are discussed. This is the first report indicating that inter-order NCH larval fish can be obtained in spite of their evolutionary divergence.
Assuntos
Peixes/embriologia , Larva/crescimento & desenvolvimento , Animais , Citoplasma , Células Híbridas , Técnicas de Transferência NuclearRESUMO
Teleosts from different families and orders were used as materials for nuclear transplantation experiments. (1) The nuclei of goldfish (Carassius auratus, family Cyprinidae, order Cypriniformes) were transplanted into the enucleated egg cytoplasm of loach (Paramisgurnus dabryanus, family Cobitidae, order Cypriniformes) and vice-versa. (2) The nuclei of Tilapia (oreochromis nilotica, order Perciformes) were transplanted into the enucleated egg cytoplasm of goldfish (Carassius auratus, order Cypriniformes). The chromosome number of the nucleus donor fish is different from that of the cytoplasmic recipient fish in each of the two combinations. In the first case, only a few early nucleo-cytoplasmic hybrid (NCH) larval fish were obtained in each combination. In second case, even though a high percentage of NCH blastulas were also obtained, the majority of them died at the same developmental stage, except a few which survived until early gastrula stage. The examination of the metaphase chromosome figures of the NCH blastulas or embryos obtained in all three combinations indicated that they were of nucleus-donor type. The developmental rates of all the NCH eggs were similar to those of cytoplasmic-recipient type. Scanning electronmicroscopy examination showed that the morphology of NCH blastula cells, which were obtained from the combination of Tilapia nucleus and goldfish cytoplasm, manifested obviously abnormal features and the cells were arrested at different stages of cell disintegration. Two-dimension polyacrylamide gel electrophoretograms of the homogenates of Tilapia, goldfish and their NCH blastula cells showed that the protein synthetic pattern of NCH blastula was similar to that of Tilapia nucleus type. The results of experiments which failed to obtain NCH adult fish in all three combinations can be explained as a result of developmental incompatibility between the donor nucleus and the enucleated recipient egg cytoplasm, which were from distantly related fish species. And the chromosome numbers of all the component fish of the three combinations which were examined in the experiment and shown to be quite different from each other in the tested fish, should not be overlooked as one of the essential factors causing the developmental incompatibility in NCH fish in this experiment.
Assuntos
Citoplasma/fisiologia , Peixes/embriologia , Técnicas de Transferência Nuclear , Óvulo/ultraestrutura , Animais , Blastocisto/fisiologia , Blastocisto/ultraestrutura , Núcleo Celular/fisiologia , Núcleo Celular/ultraestrutura , Cipriniformes/anatomia & histologia , Cipriniformes/embriologia , Citoplasma/ultraestrutura , Eletroforese em Gel Bidimensional , Feminino , Fertilização , Peixes/anatomia & histologia , Carpa Dourada/anatomia & histologia , Carpa Dourada/embriologia , Masculino , Microscopia Eletrônica de Varredura , Perciformes/anatomia & histologia , Perciformes/embriologiaRESUMO
Platelet serotonin (5-HT) studies were conducted with 12 hyperserotonemic and 12 normoserotonemic age-, sex-, and relationship-matched relatives of autistic probands. Each group consisted of 7 mothers, 4 fathers, and 1 sister of autistic children and adolescents. The density (Bmax) of platelet 5-HT2 receptor binding sites, labelled with [3H]-lysergic acid diethylamide (LSD), was significantly lower in 11 hyperserotonemic subjects compared to 12 normoserotonemic subjects (40.9 +/- 13.5 fmol/mg protein, 59.6 +/- 13.2; p < 0.004). The affinity (Kd) for [3H]-LSD binding did not differ. Although the density (Bmax) of [3H]-paroxetine binding did not differ between groups, there was a small difference in the affinity (Kd) for [3H]-paroxetine binding (hyperserotonemic 47.6 +/- 9.0 pM, normoserotonemic 54.8 +/- 12.1; p < 0.05). There were no significant differences in platelet 5-HT uptake, or in thrombin-stimulated 5-HT release. Basal, 5-HT-stimulated, and arginine-vasopressin (AVP)-stimulated inositol phosphate production, as well as basal, prostaglandin E1 (PGE1)-, and forskolin-stimulated cAMP production did not differ. There were significant correlations between whole blood 5-HT levels and LSD Bmax (rs = -0.63, N = 23, p < 0.002) and whole blood 5-HT levels and 5-HT uptake Vmax (rs = 0.56, N = 18, p < 0.02). However, [3H]-LSD labelled 5-HT2 binding and 5-HT uptake were not correlated with each other. Hyperserotonemia of autism may be heterogeneous with one subgroup of subjects with increased 5-HT uptake and another subgroup with decreased 5-HT2 binding.
Assuntos
Transtorno Autístico/sangue , Transtorno Autístico/genética , Plaquetas/metabolismo , AMP Cíclico/sangue , Receptores de Serotonina/metabolismo , Serotonina/sangue , 1-Metil-3-Isobutilxantina/farmacologia , Adulto , Alprostadil/farmacologia , Transporte Biológico , Plaquetas/efeitos dos fármacos , Criança , Colforsina/farmacologia , Feminino , Humanos , Técnicas In Vitro , Ácido Lisérgico/sangue , Masculino , Paroxetina/sangue , Contagem de PlaquetasRESUMO
This study aimed to investigate the therapeutic mechanism of treating SMMC-7721 liver cancer cells with magnetic fluid hyperthermia (MFH) using Fe2O3 nanoparticles. Hepatocarcinoma SMMC-7721 cells cultured in vitro were treated with ferrofluid containing Fe2O3 nanoparticles and irradiated with an alternating radio frequency magnetic field. The influence of the treatment on the cells was examined by inverted microscopy, MTT and flow cytometry. To study the therapeutic mechanism of the Fe2O3 MFH, Hsp70, Bax, Bcl-2 and p53 were detected by immunocytochemistry and reverse transcription polymerase chain reaction (RT-PCR). It was shown that Fe2O3 MFH could cause cellular necrosis, induce cellular apoptosis, and significantly inhibit cellular growth, all of which appeared to be dependent on the concentration of the Fe2O3nanoparticles. Immunocytochemistry results showed that MFH could induce high expression of Hsp70 and Bax, decrease the expression of mutant p53, and had little effect on Bcl-2. RT-PCR indicated that Hsp70 expression was high in the early stage of MFH (<24 h) and became low or absent after 24 h of MFH treatment. It can be concluded that Fe2O3MFH significantly inhibited the proliferation of in vitro cultured liver cancer cells (SMMC-7721), induced cell apoptosis and arrested the cell cycle at the G2/M phase. Fe2O3 MFH can induce high Hsp70 expression at an early stage, enhance the expression of Bax, and decrease the expression of mutant p53, which promotes the apoptosis of tumor cells.
Assuntos
Carcinoma Hepatocelular/tratamento farmacológico , Carcinoma Hepatocelular/terapia , Compostos Férricos/uso terapêutico , Hipertermia Induzida/métodos , Neoplasias Hepáticas/terapia , Magnetoterapia/métodos , Nanopartículas/uso terapêutico , Apoptose/efeitos dos fármacos , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Citometria de Fluxo , Hematínicos/uso terapêutico , Humanos , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Neoplasias Hepáticas/patologia , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
The following study analysed apoptosis in proliferative cells and migrating neurons of the developing cerebellum. The external granular layer, Purkinje cell layer and internal granular layer in the developing mouse cerebellar cortex were analysed by active caspase-3 immunohistochemistry, Hoechst 33258 staining and Western blot analysis. Immunocytochemistry results indicated that the peak of apoptosis appeared at postnatal days P8, P5 and P9 in the external granular layer, Purkinje cell layer and internal granular layer, respectively. Subsequently, in each region, the rate of apoptosis decreased with increasing age. In contrast, Western blot results demonstrated the highest expression of activated caspase-3 in the cerebellum at P5, followed by a subsequent decline and disappearance of expression by P14. Activated caspase-8 was expressed maximally at P10, and subsequently disappeared by P30. The results of this study suggest that the key period of neuronal apoptosis in the cerebellar cortex is between P0 and P14, indicating that this developmental period could be susceptible to treatment for congenital neurodegenerative diseases.
Assuntos
Apoptose/fisiologia , Cerebelo/embriologia , Neurônios/fisiologia , Fatores Etários , Animais , Animais Recém-Nascidos , Bisbenzimidazol , Western Blotting , Caspase 3/genética , Caspase 3/metabolismo , Caspase 8/genética , Caspase 8/metabolismo , Sistema Nervoso Central/embriologia , Imunofluorescência , Camundongos , Células de Purkinje/fisiologia , Coloração e RotulagemRESUMO
This study aimed to investigate the therapeutic mechanism of treating SMMC-7721 liver cancer cells with magnetic fluid hyperthermia (MFH) using Fe2O3 nanoparticles. Hepatocarcinoma SMMC-7721 cells cultured in vitro were treated with ferrofluid containing Fe2O3 nanoparticles and irradiated with an alternating radio frequency magnetic field. The influence of the treatment on the cells was examined by inverted microscopy, MTT and flow cytometry. To study the therapeutic mechanism of the Fe2O3 MFH, Hsp70, Bax, Bcl-2 and p53 were detected by immunocytochemistry and reverse transcription polymerase chain reaction (RT-PCR). It was shown that Fe2O3 MFH could cause cellular necrosis, induce cellular apoptosis, and significantly inhibit cellular growth, all of which appeared to be dependent on the concentration of the Fe2O3 nanoparticles. Immunocytochemistry results showed that MFH could induce high expression of Hsp70 and Bax, decrease the expression of mutant p53, and had little effect on Bcl-2. RT-PCR indicated that Hsp70 expression was high in the early stage of MFH (<24 h) and became low or absent after 24 h of MFH treatment. It can be concluded that Fe2O3 MFH significantly inhibited the proliferation of in vitro cultured liver cancer cells (SMMC-7721), induced cell apoptosis and arrested the cell cycle at the G2/M phase. Fe2O3 MFH can induce high Hsp70 expression at an early stage, enhance the expression of Bax, and decrease the expression of mutant p53, which promotes the apoptosis of tumor cells.