RESUMO
Objective: To evaluate the clinical safety and efficacy of toric intraocular lens (IOL) implantation for more than 5 years. Methods: This study was a prospective cohort study in which subjects were continuously observed over a two-year period (May 2014 to May 2016) in nine hospitals. The study randomly assigned subjects to two groups using a central dynamic randomization system: the study group, which received Proming® IQ toric IOL implants, and the control group, which received AcrySof® IQ toric IOL implants. The subjects completed a one-year follow-up, during which various measures were taken and evaluated, including visual acuity, IOL rotation, postoperative complications, intraocular pressure, and subjective evaluation (preoperatively and at 1 day, 6 months, 1 year, and 5 years post-surgery). The main statistical analysis methods include the Mann-Whitney U test, independent sample t-test, Wilcoxon signed rank test, paired sample t-test, chi-square test, and Fisher's exact test. Results: A total of 45 eyes (26 in the study group and 19 in the control group) completed the five-year continuous observation period. The mean age of the subjects was (72.07±10.67) years and the mean interval from surgery to the last visit was (5.39±0.47) years. After five years, there were no significant differences in uncorrected distance visual acuity (0.20±0.26 vs. 0.16±0.13, t=0.17,P=0.752), best corrected distance visual acuity[0.00(0.00, 0.20) vs. 0.05±0.10, U=188.00, P=0.880], uncorrected near visual acuity[0.50 (0.20, 0.60) vs. 0.42±0.20, t=0.35, P=0.857], and best corrected near visual acuity (0.13±0.16 vs. 0.17±0.23, U=161.00, P=0.884) between the two groups. However, all measures improved significantly from baseline levels in both groups (all P<0.05). Five years after surgery, no matter objective refraction [(-0.67±0.85) D vs. (-0.73±1.08)D] or subjective refraction[-0.50 (-1.00, 0.00)D vs. (0.69±0.87)D], the degree of cylindrical degree is significantly lower than preoperative corneal astigmatism [(1.27±0.49) D vs. (1.34±0.82) D, all P<0.001]. In addition, there were no significant differences in intraocular pressure, subjective evaluation of visual adverse symptoms, distance vision spectacle independence, or overall satisfaction evaluation between the two groups (all P>0.05). The IOL rotation was 3.0°(1.0°, 6.0°) in the study group and 4.0°(2.0°, 6.0°)in the control group (U=185.50,P=0.574), indicating no significant difference between the groups in terms of rotational stability. Five years after surgery, there were 7 cases of posterior capsular opacification in the study group and 4 cases in the control group. There were no cases of IOL glistening in the study group, but 5 cases (26.32%) were observed in the control group. Conclusions: The long-term effects of Proming® toric IOL implantation in correcting cataracts with regular corneal astigmatism are clear after five years, with few complications and stable results.
Assuntos
Catarata , Implante de Lente Intraocular , Idoso , Idoso de 80 Anos ou mais , Humanos , Pessoa de Meia-Idade , Astigmatismo/cirurgia , Opacificação da Cápsula/cirurgia , Doenças da Córnea/cirurgia , Implante de Lente Intraocular/efeitos adversos , Implante de Lente Intraocular/métodos , Lentes Intraoculares , Estudos Prospectivos , Refração OcularRESUMO
Objective: To understand the current situation of emergency department nurses' work engagement, and analyze the relationship between emergency department nurses' organizational climate perception, work-related acceptance actions and work engagement. Methods: In May 2021, 273 emergency department nurses from 6 class â ¢ class a general hospitals in Tianjin were selected as the research objects by using the convenient sampling method, and the general information questionnaire, work input scale, nurses' organizational climate perception scale and work-related acceptance action questionnaire were used for questionnaire survey. The correlation between job involvement and nurses' organizational climate perception and job-related acceptance action was analyzed by pearson correlation, and the influencing factors of job involvement were analyzed by multiple linear regression. Results: The average score of job involvement was (3.57±0.45) , the average score of nurses' organizational climate perception was (3.29±0.69) , and the score of work-related acceptance action was 35.00 (29.00, 47.00) . The results of correlation analysis showed that there was a positive correlation between nurses' organizational climate perception, job acceptance action and job involvement in emergency department (r=0.435, 0.518, P<0.05) . Multiple stepwise regression analysis showed that job acceptance, nurses' perception of organizational climate, health status, specialist nurses, education and emergency work years were the influencing factors of emergency department nurses' job involvement (P<0.05) , accounting for 41.9% of the total variation. Conclusion: We should create a good organizational atmosphere, improve the acceptance of emergency department nurses to nursing work, and improve the level of nurses' work input.
Assuntos
Enfermeiras e Enfermeiros , Recursos Humanos de Enfermagem Hospitalar , Serviço Hospitalar de Emergência , Humanos , Satisfação no Emprego , Percepção , Inquéritos e Questionários , Engajamento no TrabalhoRESUMO
Objective: To evaluate the value of intraoperative cerebral oxygen saturation in predicting postoperative neurocognitive dysfunction (PND) in elderly patients with mild cognitive impairment. Methods: A total of 210 cases of lumbar decompression, bone grafting and fusion surgery under general anesthesia were collected in the Third Central Hospital of Tianjin from June, 2019 to January, 2020, either sex, aged 65-75 year, BMI 19.5-32.5 kg/m(2), ASA physical status â ¡ or â ¢, preoperative comorbidities with mild cognitive impairment. MoCA and MMSE were used to evaluate the cognitive function of patients 1 day before the operation, 7 days and 3 monthes after operation. PND group (n=38) and non-PND group (n=172) were selected according to postoperative MMSE and MoCA scale scores and the diagnostic criteria of PND. Heart rate (HR) , mean arterial pressure (MAP), pulse oxygen saturation (SpO(2)), bispectral index (BIS), cerebral tissue oxygen saturation (SctO(2), average left and right brain SctO(2) were recorded) were recorded pre-anesthetic (T(0)), ten minutes of anesthesia(T(1)), twenty minutes of anesthesia (T(2)), thirty minutes into the operation (T(3)), one hour into the operation (T(4)), end of the surgery (T(5)), and leave the PACU (T(6)). SctO(2) at time point T(0) was the base value of SctO(2), and the maximum percentage drop in SctO(2) from the base value was calculated (SctO(2max)%). Results: The incidence of PND was 18% (38/210) in 210 elderly patients undergoing surgery. The age of PND group and non-PND group was (71.0±2.1) and (67.8±2.0) years old, and the PACU time was (57±5) and (46±8) min, respectively. Compared with the non-PND group, the age of the PND group was higher (t=2.600, P<0.05) and the PACU time was longer (t=3.039, P<0.05). At the time points T(3), T(4), T(5) and T(6), SctO(2) in the PND group was (62±10) %, (60±11) %, (64±12) % and (66±10)%, respectively, lower than that in the non-PND group (67±60) %, (68±6) %, (69±5) % and (70±7)%, respectively, and the difference was statistically significant (t=3.369, 4.906, 3.787, 2.516, all P<0.05).The MoCA and MMSE scores of the PND group were (22.9±1.2) and (24.1±1.2) points, respectively, 1 day before surgery; and the MoCA and MMSE scores of the PND group were reduced to (20.8±1.2) and (21.3±0.7) points, respectively, 7 days after surgery, with statistically significant differences (t=3.523, 5.675, all P<0.05). MoCA and MMSE scores 7 days after surgery in the non-PND group were (22.4±1.3) and (23.1±1.6) points, respectively. Compared with the non-PND group, MoCA and MMSE scores 7 days after surgery in the PND group were reduced (t=2.630, 3.108, all P<0.05). The critical value of intraoperative SctO(2max)% was 13.74%, the area under the curve of PND was predicted to be 0.907 (95%CI: 0.819-0.995), sensitivity and specificity were 88.9% and 88.5%, respectively. Conclusion: SctO(2max)%>13.74% can be used as an indicator to predict PND occurrence in elderly patients with mild cognitive impairment during lumbar surgery.
Assuntos
Disfunção Cognitiva , Idoso , Anestesia Geral , Cognição , Disfunção Cognitiva/etiologia , Humanos , Oxigênio , Troca Gasosa PulmonarRESUMO
Objective: To study the clinical features, continuous care and prognosis of the patients with severe and refractory anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis after intensive care unit (ICU). Methods: Clinical data of patients with severe and refractory anti-NMDAR encephalitis, who were transferred from ICU to general ward of neurology between December 2015 and October 2019, were retrospectively reviewed and analyzed in the study. Results: Twenty patients (11 females and 9 males) were enrolled in the study. The median course of disease when patients were transferred to general ward was 4.4 (2.0, 6.0) months. Six cases were alert, 6 cases were in a coma, 5 were in the early recovery phase and 3 were in the late recovery phase. Severe malnutrition, pneumonia, urinary tract infections, bedsores and leukocytopenia were common complications. Seven out of 18 patients were tested positive for cerebrospinal fluid anti-NMDAR antibodies with high titers (≥1â¶100). During this continuous therapy stage,10 patients were treated with intravenous immunoglobulin (IVIg), 1 with methylprednisolone, 2 with rituximab, 1 with intrathecal methotrexate and 1 received intravenous cyclophosphamide. All Patients were prescribed a long-term immunotherapy (mycophenolate mofetil 1.5-3.0 g/d). Sixteen patients (80%) had good prognosis (modified Rankin Scale (mRS)≤2), and the mortality was 10%, with follow-up time of 17.0 (8.0, 27.0) months. Conclusions: Patients with anti-NMDAR encephalitis, who are transferred from ICU, have severely impaired neurologic function. These patients need long-term individualized immunotherapy and continuous neurological care. Good outcomes can be achieved in most patients.
Assuntos
Encefalite Antirreceptor de N-Metil-D-Aspartato , Cuidados Críticos , Feminino , Humanos , Masculino , Prognóstico , Receptores de N-Metil-D-Aspartato , Estudos RetrospectivosRESUMO
OBJECTIVE: To investigate the clinical characteristics of rheumatoid arthritis (RA) patients with malignant tumor. METHODS: Retrospective summary was made of 1 562 in patients of RA from January 2011 to June 2017. In the study, 74 RA patients with malignant tumor were reviewed and analyzed, and the general conditions, tumor types, RA and tumor onset sequence, and the medication situation were analyzed. RESULTS: The incidence of malignant tumor in the patients with rheumatoid arthritis in our center was 4.16%. The 74 patients were complicated with malignant tumor, of whom 53 were female, and 21 male. The age of RA at presentation was (52.6±17.8) years. The average disease duration of malignant tumor was (63.4 ± 12.7) years. The onset time of rheumatoid arthritis was earlier than that of malignant tumors in 51 cases (51/74), with an average of (17.2±14.2) years between 2 and 60 years. The incidence of malignant tumor was earlier than that of rheumatoid arthritis in 16 cases (16/74), with an average of (6.2±5.9) years between 1 and 21 years, of which 10 cases were sex hormone related tumors. Seven cases (7/74) were diagnosed with RA at the same time, and the time interval between the two diseases was within 1 year. All the patients were over 60 years old with digestive tract tumors. All the 7 patients showed polyarthritis, significantly increased erythrocyte sedimentation rate and C-reactive protein, including 4 rheumatoid factor positive cases and 2 anti-CCP antibody positive cases. The effect of non-steroidal anti-inflammatory drugs and traditional drugs to improve the condition of the disease was poor in the 7 patients, and the condition was relieved after using low-dose glucocorticoids. Gastrointestinal tumors, breast and reproductive system tumors were the most common, followed by respiratory, urological and blood system tumors. CONCLUSION: The risk in patients of rheumatoid arthritis complicated with malignant tumor is higher than that of the general population. A variety of factors play an important role in cancer risk of RA, including disease activity, some estrogen metabolites, the use of drugs and so on. Therefore, all RA patients should be screened for malignant tumor during diagnosis, and malignant tumor surveillance is mandatory for all rheumatoid arthritis patients after diagnosis.
Assuntos
Artrite Reumatoide , Neoplasias , Adulto , Idoso , Artrite Reumatoide/sangue , Artrite Reumatoide/complicações , Autoanticorpos , Proteína C-Reativa/análise , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias/complicações , Neoplasias/imunologia , Peptídeos Cíclicos , Estudos Retrospectivos , Fator Reumatoide/sangueRESUMO
Objective: To evaluate the effectiveness and safety of Proming(®)Toric intraocular lens (IOL) in adults with cataract combined with corneal regular astigmatism. Methods: Multicentre, randomized, open and positive parallel controlled clinical study. A total of 121 patients (121 eyes) who had cataract combined with corneal regular astigmatism and met the inclusion criteria were enrolled in 9 hospitals from May 2014 to May 2016. There were 45 males and 76 females and the median age was 71 (42-88) years old. A total of 121 patients (121 eyes) were randomly assigned to the study group and the control group through the Central Randomization System. Sixty patients (60 eyes) of the study group were implanted with Proming(®)Toric IOL (Model: AT1BH-AT6BH) from Eyebright Medical Technology (Beijing) Co., Ltd., and 61 patients (61 eyes) of the control group were implanted with AcrySof (®)IQ Toric IOL (Model: SN6AT2-SN6AT7) from Alcon Laboratories, Inc. The visual acuity, IOL axial position, slit lamp examination, residual astigmatism and contrast sensitivity were recorded at 1 day, 1 week, 1 month, 3 months, 6 months and 1 year postoperatively. Statistical analysis was performed using χ(2) test, independent sample t test, Mann-Whitney U test, Friedman repeated measured ANOVA on ranks and non-parametric 2-factor variance analysis. Results: A total of 118 patients completed 6 months of follow-up, including 59 from the study group and 59 from the control group. The difference between the two groups in terms of the percentage of best corrected distance visual acuity (BCDVA) reaching 20/40 was 1.69% [100% (59/59) vs. 98.31% (58/59) ], and the lower limit of the 95% CI (-1.60%) was greater than -10.00%. A total of 90 patients were followed up for 1 year, including 43 patients from study group and 47 patients from control group. At 1 year after operation, the percentages of the BCDVA up to 20/40 were 97.67%(42/43) in the study group and 97.87% (46/47) in the control group, and there was no significant difference between the two groups (χ(2)=0.00, P=0.95);the percentages of the uncorrected distance visual acuity (UCDVA) up to 20/40 were 81.40%(35/43) in the study group and 82.98%(39/47) in the control group, and there was no significant difference between the two groups (χ(2)=0.04, P=0.84). At 1 year follow-up, the difference of contrast sensitivity at 18.0 c/d under the bright light, dark light, bright glare and dark glare between the two groups was not statistically significant (U=468.50, P=0.17;U=528.00, P=0.28;U=465.50, P=0.19;U=629.00, P=0.39);the difference of residual astigmatism between the two groups was not statistically significant (U=798.50, P=0.08);the difference of IOL rotation degree between the two groups was not statistically significant (U=869.00, P=0.25). There were no severe inflammatory responses nor other complications associated with IOL in both groups at each follow-up point. Conclusion: The visual quality, astigmatism correction effect, rotation stability and safety of Proming(®)Toric IOL for the treatment of cataract combined with corneal regular astigmatism is equivalent to AcrySof(®) IQ Toric IOL. (Chin J Ophthalmol, 2018, 54: 349-356).
Assuntos
Astigmatismo , Catarata , Implante de Lente Intraocular , Lentes Intraoculares , Facoemulsificação , Adulto , Idoso , Idoso de 80 Anos ou mais , Astigmatismo/terapia , Pequim , Catarata/terapia , Feminino , Humanos , Masculino , Estudos Prospectivos , Refração Ocular , Resultado do TratamentoRESUMO
microRNA (miR)-142-3p is implicated in malignancy and has been identified as a biomarker for aggressive and recurrent lung adenocarcinomas. This study aimed to evaluate the inhibitory effect of miR-142-3p on apoptosis and inflammation induced by bleomycin in MLE-12 cells. MLE-12 cells were first transfected either with miR-142-3p mimic or miR-142-3p inhibitor and then the cells were exposed to 50 µg/mL of bleomycin. Thereafter, cell viability, apoptosis and the expression of pro-inflammatory cytokines were assessed using CCK-8, flow cytometry, RT-PCR and western blot analyses. Cox-2, PI3K, AKT and mTOR expressions were detected by western blotting after bleomycin was administered together with NS-398 (an inhibitor of Cox-2). As a result, cell viability was significantly decreased, as well as apoptosis and the expression of IL-1 and TNF-α were remarkably increased after 50 and 100 µg/mL of bleomycin administration. miR-142-3p overexpression alleviated bleomycin-induced apoptosis and overproduction of these two pro-inflammatory cytokines, while miR-142-3p suppression exhibited completely opposite results. Up-regulation of Cox-2 and inactivation of PI3K/AKT/mTOR were found in bleomycin-pretreated cells, while these abnormal regulations were partially abolished by miR-142-3p overexpression and NS-398. In conclusion, this study demonstrated that miR-142-3p overexpression protected bleomycin-induced injury in lung epithelial MLE-12 cells, possibly via regulating Cox-2 expression and PI3K/AKT/mTOR signaling pathway. These findings provide evidence that miR-142-3p may be a therapeutic strategy for idiopathic pulmonary fibrosis (IPF) treatment.
Assuntos
Apoptose/efeitos dos fármacos , Bleomicina/farmacologia , Ciclo-Oxigenase 2/metabolismo , Regulação para Baixo/efeitos dos fármacos , Pulmão/citologia , MicroRNAs/metabolismo , Linhagem Celular , Humanos , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , TransfecçãoRESUMO
Human tissue kallikrein (hK1) plays an important role in regulation of blood pressure, electrolyte and glucose transport, and renal function. To evaluate the feasibility of expression of recombinant human tissue kallikrein (rhK1) in the egg whites of laying hens, human tissue kallikrein gene (hKLK1) cDNA was subcloned into the chicken oviduct-specific expression vector (pOV3), and the resultant recombinant vector pOV3K was injected into laying hens via wing vein after mixing with polyethyleneimine. Following injection twice with the recombinant vector, the enzymatic activity at a maximal level of 59 U/mL was detected in the egg whites, which lasted for more than 7 d. The expression level of rhK1 in the egg whites in the 3-mg group was relatively higher than that in the 2-mg group, but the significant differences were identified on d 7 and 8 (P < 0.05). Ten days after the primary injection, the hens were reinjected with the same dose of the vector, and even higher enzymatic activity was detected in their egg whites. Two different breeds of hen were tested with no difference in expression level found (P > 0.05). Western blot analysis of the egg whites from vector-injected hens showed the rhK1 was recognized by a polyclonal antibody specific for hK1 with molecular weights of 37 and 43 kDa, which probably corresponded to the mature and preenzyme, respectively. Biochemical studies showed that the recombinant enzyme had a similar thermostability, optimal pH, hypotensive effect, and sensitivity to different ions to the natural enzymes in human and porcine tissues. These data indicate that the chicken oviduct-specific transient expression system can produce relatively high level and authentic recombinant enzyme with a potential for further development for therapeutic use.
Assuntos
Galinhas/genética , Galinhas/fisiologia , Clara de Ovo/análise , Oviposição , Calicreínas Teciduais/genética , Calicreínas Teciduais/metabolismo , Animais , Feminino , Regulação da Expressão Gênica , Humanos , Oviductos/metabolismo , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Calicreínas Teciduais/químicaRESUMO
Human postmortem cardiac muscle was studied by immunofluorescent microscopy. Necrotic cells in acute myocardial infarctions were first identified with the hematoxylin-eosin stain as showing hypereosinophilia and autofluorescence. The results of the immunofluorescence staining showed a marked decrease if not absence of labeling for the Ca+ and Mg+ adenosine triphosphatase (ATPase) and tropomyosin in all necrotic muscle cells within a myocardial infarction. Myocytolytic cells located at the border of the infarct showed a labeling intensity similar to that of normal muscle cells. The use of immunofluorescence localization of muscle-specific proteins can be used as a reliable method to detect myocardial cell necrosis.
Assuntos
Infarto do Miocárdio/patologia , Miocárdio/patologia , Adenosina Trifosfatases/análise , ATPase de Ca(2+) e Mg(2+) , ATPases Transportadoras de Cálcio/análise , Humanos , Microscopia de Fluorescência , Infarto do Miocárdio/metabolismo , Miocárdio/metabolismo , Necrose , Tropomiosina/análiseRESUMO
Function of the largest plasmid of Yersinia pestis was studied. Assay of four strains of highly virulent Y. pestis isolated from different natural sources demonstrated that the characters of endotoxin production, sugar and alcohol fermentation, nitrate reduction as well as requirement in nutrients are mediated by no Y. pestis plasmids. On the contrary, FI and FII antigens are determined by the largest plasmid in some strains and are chromosome-mediated in others. It may well be that the genes for these antigens are comprised within the transposon-like DNA fragment.
Assuntos
Plasmídeos , Yersinia pestis/genética , Álcoois/metabolismo , Antígenos de Bactérias , Metabolismo dos Carboidratos , Elementos de DNA Transponíveis , Endotoxinas/biossíntese , Genes Bacterianos , Nitratos/metabolismo , Oxirredução , Especificidade da Espécie , Yersinia pestis/imunologiaRESUMO
microRNA (miR)-142-3p is implicated in malignancy and has been identified as a biomarker for aggressive and recurrent lung adenocarcinomas. This study aimed to evaluate the inhibitory effect of miR-142-3p on apoptosis and inflammation induced by bleomycin in MLE-12 cells. MLE-12 cells were first transfected either with miR-142-3p mimic or miR-142-3p inhibitor and then the cells were exposed to 50 μg/mL of bleomycin. Thereafter, cell viability, apoptosis and the expression of pro-inflammatory cytokines were assessed using CCK-8, flow cytometry, RT-PCR and western blot analyses. Cox-2, PI3K, AKT and mTOR expressions were detected by western blotting after bleomycin was administered together with NS-398 (an inhibitor of Cox-2). As a result, cell viability was significantly decreased, as well as apoptosis and the expression of IL-1 and TNF-α were remarkably increased after 50 and 100 μg/mL of bleomycin administration. miR-142-3p overexpression alleviated bleomycin-induced apoptosis and overproduction of these two pro-inflammatory cytokines, while miR-142-3p suppression exhibited completely opposite results. Up-regulation of Cox-2 and inactivation of PI3K/AKT/mTOR were found in bleomycin-pretreated cells, while these abnormal regulations were partially abolished by miR-142-3p overexpression and NS-398. In conclusion, this study demonstrated that miR-142-3p overexpression protected bleomycin-induced injury in lung epithelial MLE-12 cells, possibly via regulating Cox-2 expression and PI3K/AKT/mTOR signaling pathway. These findings provide evidence that miR-142-3p may be a therapeutic strategy for idiopathic pulmonary fibrosis (IPF) treatment.
Assuntos
Humanos , Bleomicina/farmacologia , Regulação para Baixo/efeitos dos fármacos , Apoptose/efeitos dos fármacos , MicroRNAs/metabolismo , Ciclo-Oxigenase 2/metabolismo , Pulmão/citologia , Transfecção , Linhagem Celular , Pulmão/efeitos dos fármacos , Pulmão/metabolismoAssuntos
Adenocarcinoma/patologia , Anticorpos Antineoplásicos/imunologia , Neoplasias Pulmonares/patologia , Peptídeos/imunologia , Linhagem Celular , Transformação Celular Neoplásica , Células Cultivadas , Receptores ErbB , Humanos , Receptores de Superfície Celular/imunologia , Fatores de Crescimento TransformadoresAssuntos
Terapia por Acupuntura , Anestesia , Cininas/sangue , Dor/induzido quimicamente , Serotonina/sangue , Animais , Cães , HumanosRESUMO
Antibodies were produced to phosphoprotein fraction, and a phosphoprotein B2 obtained by carboxyl methyl cellulose column chromatography and sodium dodecyl sulfate (SDS) - polyacrylamide gel electrophoresis as described previously (Biochem. J. 183, 147 (1979)). Production of the specific antibody was confirmed by double immunodiffusion. The phosphoprotein B2 (relative mass 68 000), which was isolated from the phosphoprotein fraction by SDS-polyacrylamide gel electrophoresis, specifically reacted with the antisera, as identified by "rocket" immunoelectrophoresis. Further characterization of the antibody to the phosphoprotein was carried out by isoelectrofocusing gel electrophoresis. The phosphoprotein, previously identified in the isoelectric point (pI) region 6.2--8.5, was subsequently reacted with antisera and 125I-labelled protein A. A prominent radioactive peak was identified in the region in which the phosphoprotein was focused. The radioimmunoactivity was proportional to the amount of phosphoproteins present in the isoelectric focusing gel. The presence of phosphoprotein antibody. The relationship between the phosphoprotein and chromatin structure, and possible role in gene regulation is discussed.
Assuntos
Nucleossomos/análise , Fosfoproteínas/imunologia , Animais , Anticorpos/análise , Proteínas Cromossômicas não Histona/análise , Imunodifusão , Focalização Isoelétrica , Masculino , Modelos Biológicos , Radioimunoensaio , RatosRESUMO
[32P]Pi was administered to rats (5mCi/rat) 2h before the isolation of liver nuclei. The isolated nuclei were subjected to mild micrococcal-nuclease digestion for 2.5, 5 and 10 min at 37 degrees C, and the mononucleosomal fraction was subsequently isolated by sucrose-density-gradient centrifugation. The specific radioactivity of 32P-labelled mononucleosomal fractions decreased with increased digestion times. A phosphorylated chromosomal protein, B2 (Mr 68000, pI6.5-8.2), was demonstrated immunologically in the mononucleosomal fraction by using an antibody specific to this electrophoretically purified phosphoprotein. The incorporation of 32P into this phosphoprotein, previously shown to be mainly through covalent linkage, was revealed by antibody precipitation followed by gel electrophoresis. The rate of release of acid-soluble nucleotides by micrococcal-nuclease digestion of liver nuclei from partially hepatectomized rats 16 h after operation was strikingly higher than that for sham-operated controls. After partial hepatectomy, an increase in 32P incorporation into phosphoprotein in the monomer fractions specifically precipitated by this antibody was also found. This suggests that the phosphorylated non-histone chromatin protein B2 is preferentially associated with the transcriptionally active chromatin.
Assuntos
Proteínas Cromossômicas não Histona/metabolismo , Nuclease do Micrococo/farmacologia , Nucleossomos/metabolismo , Fosfoproteínas/metabolismo , Animais , Centrifugação com Gradiente de Concentração , Precipitação Química , Proteínas Cromossômicas não Histona/imunologia , Eletroforese em Gel de Poliacrilamida , Hepatectomia , Técnicas In Vitro , Masculino , Nucleossomos/efeitos dos fármacos , Nucleotídeos , Fosfatos/metabolismo , Fosfoproteínas/imunologia , Ratos , Ratos EndogâmicosRESUMO
OBJECTIVES: To assess patients with different types of mutations of the beta myosin heavy chain (beta MHC) gene causing hypertrophic cardiomyopathy (HCM) and to determine the prognosis of patients according to the affected functional domain of beta MHC. DESIGN AND SETTING: Cohort study of subjects referred to an HCM clinic at an academic hospital. PATIENTS: 70 probands from the HCM clinic were screened for mutations of the beta MHC gene and 148 family members of the genotype positive probands were further assessed. The control group for the genetic studies consisted of 106 healthy subjects. MAIN OUTCOME MEASURES: Direct DNA sequencing was used to screen 70 probands for mutations of the beta MHC gene. Family members underwent genotypic and detailed clinical, ECG, and echocardiographic assessments. The survival of genotype positive subjects was evaluated according to the type of functional domain affected by the missense mutation and according to phenotypic characteristics. RESULTS: A mutation of the beta MHC gene was detected in 15 of 70 probands (21%). Of 148 family members studied in these 15 families, 74 were identified with a beta MHC defect. Eleven mutations were detected, including four novel mutations: Ala196Thr, Pro211Leu, Val404Leu, and Arg870Cys. Median survival was 66 years (95% confidence interval (CI) 64 to 77 years) in all affected subjects. There was a significant difference in survival between subjects according to the affected functional domain (p = 0.02). Significant independent predictors of decreased survival were the non-conservative (that is, associated with a change in the amino acid charge) missense mutations that affected the actin binding site (hazard ratio 4.4, 95% CI 1.6 to 11.8; p = 0.003) and those that affected the rod portion of beta MHC (hazard ratio 4.8, 95% CI 1.2 to 19.4; p = 0.03). No phenotypic characteristics were associated with decreased survival or cardiovascular morbidity. CONCLUSIONS: The type of beta MHC functional domain affected by the missense mutation is predictive of overall prognosis in HCM.