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1.
Zhonghua Yan Ke Za Zhi ; 60(8): 689-694, 2024 Aug 11.
Artigo em Chinês | MEDLINE | ID: mdl-39085159

RESUMO

Objective: To compare the accuracy of intraocular lens (IOL) power calculations using total keratometry (TK) versus standard keratometry (K) in post-corneal refractive surgery cataract patients. Methods: This retrospective case series study included 30 patients (36 eyes) with a history of laser corneal refractive surgery who underwent cataract extraction and IOL implantation at Qingdao Eye Hospital, Affiliated to Shandong First Medical University, from September 2022 to December 2023. The cohort comprised 16 males and 14 females, with an average age of (53.6±8.1) years. IOL power was calculated using the K-based Haigis-L and Barrett True-K formulas, as well as the TK-based Haigis and Barrett Universal Ⅱ formulas. Postoperative objective refraction was performed to obtain the actual refractive status of the operated eyes. The refractive prediction error (RPE) was defined as the difference between the actual spherical equivalent and the predicted refraction. The absolute value of the RPE was taken as the refractive absolute error (RAE). Differences in errors calculated by the four formulas were compared. Results: TK showed good consistency with K, with TK being on average 0.50 D lower than K. Analysis of variance revealed statistically significant differences in RPE among the four formulas (P<0.001). The RPE for the TK-based Haigis formula was (0.17±0.09) D, and for the Barrett Universal Ⅱ formula, it was (0.21±0.11) D, both significantly better than the K-based Haigis-L formula (-0.61±0.12) D and Barrett True-K formula (-0.57±0.11) D (all P<0.001). The percentage of eyes with postoperative RPE<±1.00 D was higher for the TK-based Haigis (92%, 33 eyes) and Barrett Universal Ⅱ (86%, 31 eyes) formulas compared to the TK-based Barrett True-K (75%, 27 eyes) and Haigis-L formulas (67%, 24 eyes), with statistically significant differences (P<0.05). Conclusions: Compared with K, TK improves the accuracy of IOL power calculation in post-corneal refractive surgery patients. Both the TK-based Barrett Universal Ⅱ and Haigis formulas demonstrate high accuracy.


Assuntos
Extração de Catarata , Catarata , Córnea , Implante de Lente Intraocular , Lentes Intraoculares , Refração Ocular , Humanos , Masculino , Feminino , Estudos Retrospectivos , Pessoa de Meia-Idade , Córnea/cirurgia , Implante de Lente Intraocular/métodos , Procedimentos Cirúrgicos Refrativos/métodos
2.
ESMO Open ; 9(7): 103494, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38981309

RESUMO

BACKGROUND: High tumor mutational burden (TMB) is one of the widely researched predictive biomarkers of immune checkpoint inhibitors and has been shown to be closely related with response to immunotherapy in multiple cancer types. However, for patients who have failed conventional therapy and are about to undergo immunotherapy, there is no consensus recommendation on the timing of tumor sampling for TMB analysis, and the effects of different therapies on TMB have not been clarified. This retrospective observational study aimed to investigate the heterogeneity of TMB and genomic mutation under the treatment pressure. PATIENTS AND METHODS: We retrospectively collected the available genomic and therapeutic information from 8051 samples across 15 tumor types (>50 samples/tumor) found in 30 published studies and investigated the distribution and heterogeneity of TMB under treatment across diverse cohorts. RESULTS: This integrated analysis has shown anticancer treatments increased TMB. Significant effects of treatment on TMB were more frequently observed in tumor types with lower treatment-naïve TMB, including breast, prostate, and pediatric cancers. For different cancer therapies, chemotherapy was prone to be correlated with an increased TMB in most cancer types. Meanwhile, the fraction of the TMB-high category of breast, prostate, and bladder cancers and glioma increased significantly after chemotherapy. Several actionable genes including ERS1 and NF1 in breast cancer, as well as some prognostic markers including TERT in bladder cancer and IDH1 in glioma, were significantly changed in post-chemotherapy tumors compared to treatment-naïve tumors. CONCLUSION: Our study reveals the heterogeneity of TMB under treatment across diverse cancer types and provides evidences that chemotherapy was associated with increases in TMB as well as the fraction of TMB-high category, suggesting that resampling tumor tissues for calculating post-chemotherapy TMB could be a better option for predicting the response to immunotherapy, especially for tumors with initially low TMB.


Assuntos
Biomarcadores Tumorais , Mutação , Neoplasias , Feminino , Humanos , Biomarcadores Tumorais/genética , Genômica/métodos , Imunoterapia/métodos , Neoplasias/genética , Neoplasias/tratamento farmacológico , Estudos Retrospectivos
3.
Zhonghua Yan Ke Za Zhi ; 59(12): 1012-1018, 2023 Dec 11.
Artigo em Chinês | MEDLINE | ID: mdl-38061902

RESUMO

Objective: To evaluate the accuracy of five intraocular lens (IOL) power calculation formulas for calculating IOL power in patients with previous myopia-corrected corneal refractive surgery. Methods: In this case series study, a total of 30 eyes of 30 patients who had excimer laser corneal refractive surgery for myopia and subsequent cataract surgery in Qingdao Eye Hospital from April 2020 to October 2022 were included. The Pentacam anterior segment analysis system and IOLMaster were used to measure ocular parameters, including axial length, anterior chamber depth, keratometry, lens thickness, and mean true net power (mTNP). Five formulas were used for IOL power calculation: Shammas formula, Olsen formula, SRK/T (mTNP) formula, Haigis-L formula, and Barrett True-K formula. After cataract extraction, we obtained the actual postoperative refraction by measuring the objective refraction. The prediction error was determined as the difference between the actual postoperative refraction and the predicted refraction, and the absolute value of the prediction error was the absolute error. The differences in the calculation errors of the 5 formulas were compared. Results: Regarding the prediction errors, the results of the SRK/T (mTNP) and Olsen formulas were better than those of Shammas and Haigis-L, and the differences were statistically significant (all P<0.05). The proportion of eyes with an absolute error of 0.50 D for Barrett True-K was highest (70%, 21/30), followed by the SRK/T (mTNP) formula (67%, 20/30). The proportions of eyes with an absolute error within 1.00 D for Barrett True-K, SRK/T (mTNP), and Olsen were all over 80%, with 24 eyes, 24 eyes, and 25 eyes, respectively. Conclusions: The Barrett True-K formula showed high accuracy in predicting the refraction after cataract extraction in patients with a history of corneal refractive surgery for myopia. The calculation result of the Haigis-L formula was highly unstable.


Assuntos
Lentes Intraoculares , Miopia , Facoemulsificação , Procedimentos Cirúrgicos Refrativos , Humanos , Implante de Lente Intraocular , Refração Ocular , Miopia/cirurgia , Estudos Retrospectivos , Biometria , Facoemulsificação/métodos
4.
Zhonghua Yi Xue Za Zhi ; 103(48): 3924-3931, 2023 Dec 26.
Artigo em Chinês | MEDLINE | ID: mdl-38129169

RESUMO

Objective: To explore the efficacy of myocardial protection with single-dose histidine-tryptophan-ketoglutarate (HTK) cardioplegia during aortic root operation, and the correlation between short-term clinical outcomes and duration of myocardial ischemia. Methods: The data of clinical cases undergoing myocardial protection with single-dose HTK cardioplegia during aortic root operation from January 2018 to December 2022 were retrospectively reviewed. Patients were divided into conventional HTK cardioplegia group (<3 h) and prolonged HTK cardioplegia group (≥3 h) according to duration of intraoperative myocardial ischemia. A 1∶1 propensity score matching was performed and the correlations between duration of myocardial ischemia and postoperative short-term outcomes (30-day mortality, readmission, mechanical circulation support and renal insufficiency) were analyzed. Results: A total of 282 patients were included in the final analysis, with 210 cases in the conventional HTK cardioplegia group and 72 cases inthe prolonged HTK cardioplegia group before matching. After matching, there were 64 cases (53 males and 11 females) in the conventional HTK cardioplegia group, with a mean age of (49.4±14.2) years. The prolonged HTK cardioplegia group had 64 cases (55 males and 9 females), with a mean age of (50.5±12.3) years. Higher sensitivity troponin [12 h: 10.1 (4.6, 18.7) µg/Lvs 4.1(2.2, 8.6) µg/L, P=0.002; 24 h: 7.7 (4.5, 19.0) µg/L vs 4.8 (2.2, 11.9) µg/L, P=0.025] and creatine kinase isoenzyme[12 h: 46.3 (28.1, 62.4) µg/L vs 20.7(14.1, 32.9) µg/L, P<0.001; 24 h: 26.3(13.4, 49.2) µg/L vs 14.5 (10.1, 33.5)µg/L, P=0.011] after surgery was detected in prolonged HTK cardioplegia group. Comparisons of other primary and secondary endpoint events showed no significant differences between the two groups (all P>0.05). Multivariate binary logistic regression showed that duration of myocardial ischemia had no significant effect on postoperative 30-day mortality (OR=1.255, 95%CI: 0.500-3.148, P=0.629), 30-day readmission (OR=0.378, 95%CI: 0.069-2.065, P=0.261) and mechanical circulation support (OR=0.991, 95%CI: 0.331-2.970, P=0.998). Conclusion: During aortic root surgery, single-dose HTK cardioplegia may provide satisfactory myocardial protection, and there was no significant correlation between duration of myocardial ischemia and short-term clinical outcomes.


Assuntos
Doença da Artéria Coronariana , Isquemia Miocárdica , Masculino , Feminino , Humanos , Adulto , Pessoa de Meia-Idade , Histidina , Triptofano , Estudos Retrospectivos , Aorta Torácica , Soluções Cardioplégicas/uso terapêutico , Glucose , Parada Cardíaca Induzida , Manitol
6.
Zhonghua Yan Ke Za Zhi ; 59(2): 118-128, 2023 Feb 11.
Artigo em Chinês | MEDLINE | ID: mdl-36740441

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 Ocular
7.
Pediatr Blood Cancer ; 68(12): e29281, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34596937

RESUMO

BACKGROUND: High-risk pediatric acute myeloid leukemia confers a poor prognosis, and alternative strategies are needed to improve outcomes. We hypothesized that intensifying induction on the AAML1031 clinical trial would improve outcomes compared to the predecessor trial AAML0531. METHODS: Patients on AAML0531 received cytarabine (1600 mg/m2 )/daunorubicin (150 mg/m2 )/etoposide (ADE) for induction II and patients on AAML1031 received mitoxantrone (48 mg/m2 )/cytarabine (8000 mg/m2 ) (MA). Stem cell transplant (SCT) conditioning included busulfan/cyclophosphamide on AAML0531, whereas AAML1031 used busulfan/fludarabine and liberalized donor eligibility. Patients were included in this analysis if they met high-risk criteria common to the two trials by cytogenics or poor disease response after induction I ADE. RESULTS: MA provided no benefit over ADE at: induction II response (complete response [CR]: 64% vs. 62%, p = .87; measurable residual disease [MRD]+: 57% vs. 46%, p = .34); or intensification I response (CR: 79% vs. 94%, p = .27; MRD+: 27% vs. 20%, p = 1.0). When considered with altered SCT approach, MA did not improve 5-year disease-free survival (24% ± 9% vs. 18% ± 15%, p = .63) or 5-year overall survival (35% ± 10% vs. 38% ± 18%, p = .66). MA was associated with slower neutrophil recovery (median 34 vs. 27 days, p = .007) and platelet recovery (median 29 vs. 24.5 days, p = .04) and longer hospital stay (32 vs. 28 days, p = .01) during induction II. CONCLUSION: Intensification of induction II did not improve treatment response or survival, but did increase toxicity and resource utilization. Alternative strategies are urgently needed to improve outcomes for pediatric patients with high-risk acute myeloid leukemia (trials registered at clinicaltrials.gov NCT01371981, NCT00372593).


Assuntos
Quimioterapia de Indução , Leucemia Mieloide Aguda , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Bussulfano/uso terapêutico , Criança , Ensaios Clínicos como Assunto , Citarabina/uso terapêutico , Intervalo Livre de Doença , Humanos , Leucemia Mieloide Aguda/tratamento farmacológico , Leucemia Mieloide Aguda/etiologia , Neoplasia Residual/tratamento farmacológico , Resultado do Tratamento
8.
Zhonghua Yan Ke Za Zhi ; 57(7): 512-518, 2021 Jul 11.
Artigo em Chinês | MEDLINE | ID: mdl-34256471

RESUMO

Objective: To analyze the cause of late postoperative opacification of hydrophilic acrylic intraocular lenses (IOLs) and the effect of IOL replacement surgery. Methods: This retrospective case series study comprised 15 eyes of 15 patients who were diagnosed as late postoperative opacification of hydrophilic acrylic IOLs from January 2019 to June 2020 at Qingdao Eye Hospital of Shandong First Medical University. The clinical data of patients were reviewed, and two IOLs were examined by scanning electron microscopy and energy dispersive X-ray spectroscopy. The intraoperative and postoperative complications of IOL replacement surgery were evaluated, and best corrected visual acuity was compared before and after surgery. Preoperative and postoperative data were compared using the paired t test. Results: The interval between the first IOL implantation and the detection of IOL opacification in 15 patients was (27.3±5.9) months. All the 15 patients had unilateral IOL opacification, and 9 patients had hypertension. Glycosylated hemoglobin A1c was significantly higher than the normal value in 4 of the 10 patients who underwent cataract surgery at our hospital. Confocal microscopy disclosed coralliform deposits on the superficial IOL optic. Scanning electron microscopy and energy dispersive X-ray spectroscopy showed the presence of calcium and phosphorus crystals in the opacification region of IOLs. Visual acuity in all 13 eyes receiving IOL exchange was significantly improved from 1.03±0.64 (logarithm of the minimum angle of resolution) to 0.39±0.21 (P<0.05). Posterior capsule rupture (4 eyes), new IOL implanted in the ciliary sulcus (3 eyes) and zonule breaking (1 eye) occurred during IOL replacement. Conclusions: IOL opacification is related with the IOL material and calcium ion concentration on the IOL surface. IOL replacement surgery can improve visual acuity safely and effectively. (Chin J Ophthalmol, 2021, 57: 512-518).


Assuntos
Extração de Catarata , Lentes Intraoculares , Facoemulsificação , Humanos , Implante de Lente Intraocular , Complicações Pós-Operatórias/etiologia , Estudos Retrospectivos , Acuidade Visual
9.
Zhonghua Shao Shang Za Zhi ; 37(3): 243-249, 2021 Mar 20.
Artigo em Chinês | MEDLINE | ID: mdl-33618447

RESUMO

Objective: To investigate the influencing factors and their predictive value of skin graft survival after Meek grafting in severe burn patients. Methods: A retrospective case-control study was conducted in 115 severe burn patients (95 males, 20 females, aged 1-74 years) who met the inclusion criteria and received Meek grafting in the First Affiliated Hospital of Army Medical University (the Third Military Medical University) from January 2013 to December 2019. The patients were divided into good skin graft survival group with skin graft survival rate≥70% (68 cases) and poor skin graft survival group with skin graft survival rate<70% (47 cases). The statistics of patients in the two groups were recorded during their first Meek grafting after admission including the gender, age, body mass index, full-thickness burn area, burn index, complication of inhalation injury, time from injury to operation, preoperative cystatin C level, preoperative albumin level, preoperative neutrophil, preoperative hemoglobin level, preoperative platelet count, and platelet count on the first, third, and fifth day after operation. The above indicators were statistically analyzed between the two groups with independent sample t test, Mann-Whitney U test, and chi-square test. A 1∶1 propensity score matching (PSM) of the gender, age, body mass index, full-thickness burn area, burn index, complication of inhalation injury, time from injury to operation of patients in the two groups were performed to eliminate the differences in baseline data, and then the above indicators of the remaining patients in the two groups were recorded and analyzed again. The indicators with statistically significant differences between the two groups after 1∶1 PSM were selected for multivariate logistic regression analysis to screen the independent risk factors affecting the skin graft survival after Meek grafting in severe burn patients. The receiver operating characteristic (ROC) curve of independent risk factors for predicting poor skin graft survival after Meek grafting in severe burn patients after 1∶1 PSM was drawn, and the area under the curve, the cut-off value, and the sensitivity and specificity under the cut-off value were calculated. The patients after 1∶1 PSM were divided into independent risk factor>the cut-off value group and independent risk factor≤the cut-off value group with the incidence of poor skin graft survival after Meek grafting compared using the chi-square test, and the relative risk of poor skin graft survival after Meek grafting was calculated. Results: Before 1∶1 PSM, there were no statistically significant differences in gender, age, body mass index, complication of inhalation injury, time from injury to operation, preoperative cystatin C level, preoperative albumin level, preoperative neutrophil, preoperative hemoglobin level of patients between the two groups (P>0.05); the full-thickness burn area and burn index of patients in poor skin graft survival group were significantly higher than those in good skin graft survival group (Z=-2.672, -2.882, P<0.01); the preoperative platelet count and the platelet count on the first, third, and fifth day after operation of patients in poor skin graft survival group were significantly lower than those in good skin graft survival group (Z=-3.411, -3.050, -2.748, -2.686, P<0.01). After 1∶1 PSM, 46 cases were remained in each group. There were no statistically significant differences in gender, age, body mass index, full-thickness burn area, burn index, complication of inhalation injury, time from injury to operation, preoperative cystatin C level, preoperative albumin level, preoperative neutrophil, preoperative hemoglobin level of remaining patients between the two groups (P>0.05); the preoperative platelet count and the platelet count on the first, third, and fifth day after operation of patients in poor skin graft survival group were significantly lower than those in good skin graft survival group (Z=-3.428, -2.940, t=-2.427, -2.316, P<0.05 or P<0.01). Multivariate logistic regression analysis showed that the preoperative platelet count was the only independent risk factor affecting the skin graft survival after Meek grafting in severe burn patients (odds ratio=0.994, 95% confidence interval=0.989-0.998, P<0.01). The area under the ROC curve of preoperative platelet count predicting poor skin graft survival after Meek grafting in 92 patients was 0.707 (95% confidence interval=0.603-0.798, P<0.01), and the cut-off value of preoperative platelet count was 98×109/L, with sensitivity of 54.3% and specificity of 78.3% under the cut-off value. The incidence of poor skin survival after Meek grafting of patients in preoperative platelet count≤98×109/L group was 71.4% (25/35), which was obviously higher than 36.8% (21/57) in preoperative platelet count>98×109/L group (χ2=10.376, P<0.01). Compared with that in preoperative platelet count>98×109/L group, patients in preoperative platelet count≤98×109/L group had a relative risk of poor skin graft survival after Meek grafting of 2.211 (95% confidence interval=1.263-3.870). Conclusions: Preoperative platelet count is an independent risk factor affecting the skin graft survival after Meek grafting in severe burn patients and has a good predictive value. Meek grafting should be performed with caution when the preoperative platelet count of patients is≤98×109/L.


Assuntos
Queimaduras , Sobrevivência de Enxerto , Adolescente , Adulto , Idoso , Queimaduras/cirurgia , Estudos de Casos e Controles , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Prognóstico , Curva ROC , Estudos Retrospectivos , Transplante de Pele , Adulto Jovem
10.
Zhonghua Shao Shang Za Zhi ; 37(1): 9-16, 2021 Jan 20.
Artigo em Chinês | MEDLINE | ID: mdl-33499564

RESUMO

Objective: To explore the effects of B-cell lymphoma-2/adenovirus E1B 19 000 interacting protein 3 (BNIP3) on the migration and motility of human dermal microvascular endothelial cells (HDMECs) under hypoxia and the mechanism. Methods: The experimental research method was applied. (1) HDMECs were divided into normoxia group received routine culture and hypoxia 6, 12, 24 h groups treated under hypoxia with oxygen volume fraction of 2% for corresponding time according to the random number table (the same grouping method below). Western blotting was used to detect the protein expressions of BNIP3 and microtubule-associated protein 1 light chain 3Ⅱ (LC3Ⅱ) in HDMECs. (2) HDMECs were divided into normoxia+ unloaded group, normoxia+ BNIP3 knockdown group, hypoxia+ unloaded group, and hypoxia+ BNIP3 knockdown group which were transfected with unloaded virus or BNIP3 knockdown virus and were subjected to normoxic or hypoxic treatment. The BNIP3 protein expression was detected by Western blotting and immunofluorescence staining. The scratch area at 24 h post scratching was detected by scratch test, and the healing rate of scratch was calculated. The curve distance of cell movement was measured with the living cell workstation, and the speed of movement was calculated within 3 hours. (3) HDMECs were grouped and treated as experiment (2). Western blotting and immunofluorescence staining were performed to detect the protein expression of LC3Ⅱ. The number of sample was 3 in the above-mentioned experiments. Data were statistically analyzed with one-way analysis of variance and least significant difference test. Results: (1) Compared with those of normoxia group, the protein expressions of BNIP3 and LC3Ⅱ of cells in hypoxia 6, 12, 24 h groups were significantly increased (P<0.01). (2) After 6 hours of culture, compared with that of hypoxia+ unloaded group, the BNIP3 protein expressions of cells in normoxia+ unloaded group and hypoxia+ BNIP3 knockdown group were significantly decreased (P<0.05 or P<0.01). The red fluorescence denoting BNIP3 protein expression of cells in normoxia+ unloaded group and normoxia+ BNIP3 knockdown group was weak, the red fluorescence of cells in hypoxia+ unloaded group was strong, and the red fluorescence of cells in hypoxia+ BNIP3 knockdown group was significantly decreased compared with that in hypoxia+ unloaded group. After scratching for 24 hours, the scratch of cells in hypoxia+ unloaded group basically healed, while the remaining scratch area in the other three groups were large. The healing rates of scratch of cells in normoxia+ unloaded group, normoxia+ BNIP3 knockdown group, hypoxia+ unloaded group, and hypoxia+ BNIP3 knockdown group were (61±4)%, (58±4)%, (88±4)%, and (57±4)%, respectively. The healing rate of scratch of cells in hypoxia+ unloaded group was significantly higher than that in normoxia+ unloaded group (P<0.01) and hypoxia+ BNIP3 knockdown group (P<0.05). Within 3 hours of observation, the range of cell movement in hypoxia+ unloaded group was significantly larger than that in normoxia+ unloaded group, the range of cell movement in hypoxia+ BNIP3 knockdown group was significantly smaller than that in hypoxia+ unloaded group, and the curve movement velocity of cells in hypoxia+ unloaded group was significantly higher than that in normoxia+ unloaded group and hypoxia+ BNIP3 knockdown group (P<0.01). (3) After 6 hours of culture, compared with hypoxia+ unloaded group, the LC3Ⅱ protein expressions of cells in hypoxia+ unloaded group and hypoxia+ BNIP3 knockdown group were decreased significantly (P<0.05 or P<0.01). After 6 hours of culture, the red fluorescence denoting LC3 protein expressions of cells was weak in normoxia+ unloaded group and normoxia+ BNIP3 knockdown group, the red fluorescence of cells was significantly enhanced in hypoxia+ unloaded group, and the red fluorescence of cells was significantly inhibited in hypoxia+ BNIP3 knockdown group. Conclusions: BNIP3 can promote the migration and motility of HDMECs under hypoxia, and autophagy may be involved in the regulation migration of HDMECs by BNIP3.


Assuntos
Adenoviridae , Células Endoteliais , Movimento Celular , Humanos , Hipóxia , Proteínas Proto-Oncogênicas c-bcl-2
11.
Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi ; 55(11): 1043-1049, 2020 Nov 07.
Artigo em Chinês | MEDLINE | ID: mdl-33210884

RESUMO

Objective: To investigate the consistency of Velum, Oropharygneal, Tongue base, Epiglottis (VOTE) scores between two surgeons with similar clinical experience in obstructive sleep apnea hypopnea syndrome (OSAHS) patients with different degree of disease, and to analyze the influencing factors leading to the difference in score. Methods: This was a cross-sectional study. 64 preoperative drug-induced sleep endoscopy (DISE) videos of OSAHS patients during December 2014 to July 2018, from Nanfang Hospital, Southern Medical University were analyzed. The VOTE score was assessed single-blind by two similar experienced surgeons, and the Kappa value between the two scorers was calculated by the third researcher. According to the characteristics of the case, Fisher's exact test or chi-square test method was used to further explore the factors that influenced the consistency. Results: Sixty-four patients were divided into four groups according to the severity of the disease, including mild (7 cases), moderate (30 cases), severe(18 cases), and extremely severe (9 cases). The scores evaluated between two researchers were analysed for consistency. For mild patients, the two scorers were completely consistent in the configuration and degree of obstruction in the velum and epiglottis (Kappa=1). There was no agreement on whether obstruction or not, obstructed configuration, obstructed degree of the oropharynx and tongue base, and presence of velum and epiglottis obstruction. For moderate patients, the two scorers had a good consistency in the configuration and degree of the velum (0.61≤Kappa≤0.80), and there was no consistency in the evaluation of the degree of tongue base and epiglottis (P>0.05). The consistency of the remaining obstructed conditions in the four planes was generally or moderate (0.21≤Kappa≤0.60). For patients with severe OSAHS, the two raters were completely consistent in the evaluation of palatopharyngeal and epiglottic planes for the presence of obstruction, but there was no consistency in the degree of obstruction. Although the degree of obstruction in the oropharyngeal plane can be assessed with good consistency, the consistency of whether the plane was blocked or not was generally not high. In the assessment of other obstructive conditions in the four planes of severe patients, the agreement between the two scorers was moderate or generally. For extremely severe patients, the two scorers were completely consistent in the evaluation of the velum obstruction, but there was no consistency in the degree of obstruction of the oropharynx and tongue base, and the obstruction configuration and degree of the epiglottis. The evaluation of other obstructed conditions in the four planes is good or moderate. Among the patients with severe OSAHS, the difference in the assessment of obstruction of the oropharynx was associated with tonsil size (P<0.05). Conclusion: When physicians with similar clinical experience scored VOTE, the consistency of whether the velum and oropharyngeal planes are obstructed is related to the severity of the disease. Better consistency is observed among more severe OSAHS patients. The reason for the poor consistency of the oropharyngeal plane in severe OSAHS patients OSAHS is due to the difference of the tonsils size. For severe OSAHS patients with small tonsils, the assessment of whether the oropharynx is obstructed should be more cautious.


Assuntos
Epiglote , Preparações Farmacêuticas , Estudos Transversais , Endoscopia , Humanos , Método Simples-Cego , Sono , Língua
13.
Zhonghua Shao Shang Za Zhi ; 36(8): 651-657, 2020 Aug 20.
Artigo em Chinês | MEDLINE | ID: mdl-32829603

RESUMO

Objective: To investigate the effects and mechanism of mitochondrial transcription factor A (TFAM) and cytochrome c oxidase (COX) pathway in the energy production of hypoxic cardiomyocytes of rats regulated by tumor necrosis factor receptor associated protein 1 (TRAP1). Methods: The cardiomyocytes were isolated from 135 neonatal Sprague-Dawley rats (aged 1-3 d) and cultured for the following experiments. (1) Cells were collected and divided into normoxia blank control (NBC) group, hypoxia blank control (HBC) group, hypoxia+ TRAP1 over-expression control (HTOC) group, and hypoxia+ TRAP1 over-expression (HTO) group according to the random number table (the same grouping method below), with 1 bottle in each group. Cells in NBC group were cultured routinely, cells in HBC group were cultured in hypoxic condition for 6 hours after routine culture, cells in HTOC and HTO groups were respectively added with TRAP1 over-expression empty virus vector and TRAP1 over-expression adenovirus vector virus suspension for transfection for 48 hours after routine culture and then cultured in hypoxic condition for 6 hours. The protein expression of TFAM of cells in each group was detected by Western blotting. (2) Cells were collected and divided into NBC, HBC, HTOC, HTO, HTO+ TFAM interference control (HTOTIC), and HTO+ TFAM interference (HTOTI) groups, with 1 well in each group. Cells in the former 4 groups were dealt with the same methods as the corresponding groups in experiment (1). Cells in HTOTIC and HTOTI groups were respectively added with TFAM interference empty virus vector and TFAM interference adenovirus vector virus suspension for transfection for 48 hours, and the other processing methods were the same as those in HTO group. The content of ATP of cells in each group was determined by ATP determination kit and microplate reader, and the COX activity of cells in each group was determined by COX activity assay kit and microplate reader. (3) Cells were collected and divided into NBC group, normoxia+ sodium azide (NSA) group, HBC group, and hypoxia+ sodium azide (HSA) group, with 1 well in each group. Cells in NBC and HBC groups were respectively dealt with the same methods as the corresponding groups in experiment (1). Cells in NSA and HSA groups were respectively added with 32 nmol sodium azide at 30 min before experiment or hypoxia, and then cells in HSA group were cultured in hypoxic condition for 6 hours. The content of ATP was determined by the same method as above. The above three experiments were repeated for three times. Data were statistically analyzed with one-way analysis of variance and least significant difference test. Results: (1) Compared with that in NBC group, the protein expression of TFAM of cells in HBC group was significantly decreased (P<0.01). Compared with that in HBC group or HTOC group, the protein expression of TFAM of cells in HTO group was significantly increased (P<0.01). (2) Compared with 0.552±0.041 and 1.99±0.15 in NBC group, the COX activity (0.270±0.044) and ATP content (1.09±0.11) of cells in HBC group were significantly decreased (P<0.01). Compared with 0.269±0.042 and 1.17±0.12 in HBC group and those in HTOC group, the COX activity (0.412±0.032 and 0.404±0.016) and ATP content (1.75±0.06 and 1.69±0.07) of cells in HTO and HTOTIC groups were significantly increased (P<0.01). Compared with those in HTO and HTOTIC groups, the COX activity (0.261±0.036) and ATP content (1.23±0.07) of cells in HTOTI group were significantly decreased (P<0.01). (3) Compared with that in NBC group, the ATP content of cells in NSA and NBC groups was significantly decreased (P<0.01). Compared with that in HBC group, the ATP content of cells in HSA group was significantly decreased (P<0.01). Conclusions: TRAP1 can increase the COX activity of cardiomyocytes by raising the expression of TFAM, and finally alleviate the impairment in energy production of cardiomyocytes caused by hypoxia.


Assuntos
Miócitos Cardíacos , Animais , Proteínas de Ligação a DNA , Complexo IV da Cadeia de Transporte de Elétrons , Proteínas de Choque Térmico HSP90 , Hipóxia , Proteínas Mitocondriais , Ratos , Ratos Sprague-Dawley , Receptores do Fator de Necrose Tumoral , Fatores de Transcrição
14.
Neoplasma ; 67(4): 922-932, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32386484

RESUMO

With the increasing number of elderly patients, the risk of diseases such as colorectal cancer (CRC) has increased. The objective of this prospective study was to explore the effects of sarcopenia, hypoalbuminemia, and laparoscopic surgery on postoperative complications among elderly patients who recently underwent colorectal surgery. Patients aged over 65 years who underwent surgery for CRC at the First Affiliated Hospital of Wenzhou Medical University were considered for this study. The demographical and clinical characteristics of these patients, as well as postoperative complications, were prospectively analyzed. The patients were divided into two groups depending on the diagnosis of sarcopenia, and the clinical variables corresponding to the two groups were compared. Further, the risk factors associated with postoperative complications were evaluated using univariate analysis and multivariate logistic regression analysis. A total of 360 patients fulfilled the inclusion criteria. Incidences of postoperative complications in the sarcopenia and non-sarcopenia groups were at 38.3% and 27.3%, respectively. In addition, sarcopenia (p=0.029) and hypoalbuminemia (p=0.010) were identified as independent risk factors, while laparoscopic surgery (p=0.023) was identified as a protective factor for postoperative complications. However, laparoscopic surgery was a protective factor for postoperative complications in the colon group only (p=0.001). Sarcopenia and hypoalbuminemia are independent risk factors that influence the probability of developing complications following CRC surgery. Laparoscopic surgery is a protective factor for postoperative complications of CRC patients, particularly colon cancer patients.


Assuntos
Neoplasias do Colo , Neoplasias Colorretais , Hipoalbuminemia , Laparoscopia , Sarcopenia , Idoso , Neoplasias Colorretais/complicações , Neoplasias Colorretais/cirurgia , Humanos , Hipoalbuminemia/complicações , Complicações Pós-Operatórias/epidemiologia , Estudos Prospectivos , Estudos Retrospectivos , Fatores de Risco , Sarcopenia/complicações , Resultado do Tratamento
15.
Zhonghua Yan Ke Za Zhi ; 56(5): 343-348, 2020 May 11.
Artigo em Chinês | MEDLINE | ID: mdl-32450666

RESUMO

Objective: To observe the preliminary clinical effect of intraocular lens optic capture through posterior continuous curvilinear capsulorhexis in the treatment of pediatric cataract. Methods: It was a retrospective case series study. Forty-three eyes of 28 children underwent posterior continuous curvilinear capsulorhexis with posterior chamber intraocular lens optic capture to treat cataract from June 2017 to October 2018 in Qingdao Eye Hospital. Postoperative best corrected visual acuity, diopters, intraocular pressure, the position of intraocular lens, and postoperative complications were assessed. The distribution of preoperative and postoperative best corrected visual acuity was analyzed by Fisher's exact probability test. Results: Twenty-eight patients were 14 females and 14 males aged from 2 years old to 12 years old [mean age, (7±4) years]. All intraocular lenses were successfully captured in the posterior capsule. Patients were followed-up for 6.0 to 12.0 months (mean, 8.4 months). Except 2 eyes from one uncooperative child, the distribution of preoperative and postoperative best corrected visual acuity (<0.1, 0.1-<0.3, 0.3-<0.5, ≥0.5) had a significant difference (17, 17, 4, 3 eyes vs. 4, 4, 5, 28 eyes, P<0.01). At the last postoperative follow-up visit, the mean spherical equivalent was (0.21±0.74) D. Transient intraocular hypertension occurred in 3 eyes at 1 week after surgery and was controlled with stopping the use of corticosteroid eyedrops. No visual axis opacification or intraocular lens decentration or tilt was observed during the follow-up period. No other complications such as iris synechia, secondary glaucoma, retinal detachment, and cystoid macular edema were observed. Conclusions: Posterior continuous curvilinear capsulorhexis with intraocular lens optic capture is a safe and effective technique to treat pediatric cataract. It has a significant effect on the prevention of visual axis opacification after cataract surgery in children. (Chin J Ophthalmol, 2020, 56: 343-348).


Assuntos
Capsulorrexe , Extração de Catarata , Catarata , Lentes Intraoculares , Criança , Pré-Escolar , Feminino , Seguimentos , Humanos , Implante de Lente Intraocular , Masculino , Complicações Pós-Operatórias , Estudos Retrospectivos
16.
Zhonghua Yi Xue Za Zhi ; 100(12): 904-909, 2020 Mar 31.
Artigo em Chinês | MEDLINE | ID: mdl-32234164

RESUMO

Objective: To explore the mechanism of CD38-mediated cardiac damage under hypoxic-ischemic (H/I) conditions. Methods: Twenty CD38(-/-) male mice (8-week-old) and 20 wild-type (WT) male C57BL/6J mice (8-week-old) were randomly selected to construct the model of approximately 25% of the total body surface area (TBSA) burn injury. The cardiomyocytes (CMs) were separated from neonatal mice (1day) to construct the H/I injury model. Ad-CD38 adenovirus was transfected into CD38(-)/- primary CMs to callback CD38 expression. Animal experiments were grouped into WT-control group, CD38(-/-)-control group, WT-burn group, and CD38(-/-)-burn group (10 mice in each group). Primary CMs were divided into 6 groups: WT-normoxia group, CD38(-/-)-normoxia group, CD38(-/-)+Ad-CD38-normoxia group, WT-H/I group, CD38(-/-)-H/I group, CD38(-/-)+Ad-CD38-H/I group. The release of lactic dehydrogenase (LDH) from CMs and the cell viability were measured to estimate the level of myocardial injury. Ultrastructure of cardiomyocytes was examined by electron microscope. CD38 protein level and mitochondrial apoptosis-related proteins were detected by Western blot. Flow cytometry was used to detect mitochondrial reactive oxygen species (MitoSOX) of CMs under H/I condition. Cardiac function of mice was detected by ultrasonic apparatus. Results: (1) Animal experiments: The expression level of cardiac CD38 in WT-burn group was significantly higher than that in sham group (P<0.001). The heart function of CD38(-/-)-burn group was obviously better than WT-burn group [ejection fraction (EF)%: (84.70±2.31)% vs (76.10±2.96)%, shortening fraction (FS)%: (48.90±5.00)% vs (38.10±2.80)%] (both P<0.001). (2) Cell experiments: The expression level of cardiac CD38 in WT CMs under H/I condition was significantly higher than that in WT CMs under normoxia condition (P<0.05). The level of LDH, apoptotic cell and MitoSOX in CD38(-/-)-H/I group were fewer than WT-H/I group and CD38(-/-)+Ad-CD38(-)H/I group [(11.2±3.0)% vs (18.2±3.4)% and (17.6±4.0)%, (13.0±2.8)% vs (23.1±4.9)% and (23.3±6.0)%, (162±11)% vs (228±18)% and (220±18)%] (all P<0.001). The levels of cleaved-caspase3, Cytochrome-C in CD38(-/-)-H/I group were significantly lower than those in WT-H/I group and CD38(-/-)+Ad-CD38-H/I group (P<0.001). The cell viability in CD38(-/-)-H/I group was higher than that in WT-H/I group and CD38(-/-)+Ad-CD38-H/I group (0.355±0.043 vs 0.280±0.051 and 0.291±0.024) (all P<0.05). Electron microscopy results showed that structure of mitochondria in CD38(-/-)-H/I group was better than in WT-H/I group and CD38(-/-)+Ad-CD38-H/I group. Conclusion: Overexpression of CD38 contributes to cardiac damage by stimulating mitochondrial apoptotic pathway.


Assuntos
Hipóxia , Animais , Apoptose , Queimaduras , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mitocôndrias , Miócitos Cardíacos
17.
Artigo em Chinês | MEDLINE | ID: mdl-31623039

RESUMO

Objective:To analyze the related factors of VOTE score for drug-induced endoscopy(DISE) in patients with obstructive sleep apnea (OSA). Method:Fifty-four OSA patients, diagnosed by polysomnograph, underwent surgical treatment from Nov 2014 to Dec 2016 in our hospital. All patients underwent drug induced sleep endoscope, and then the collapse of pharyngeal space was evaluated. We analyzed the related factors with VOTE score. Result:The occlusion rates were significant statistical different in different spaces of 54 OSA patients undergoing DISE(P=0.000, velum 98.15%, oropharynx 81.48%, tongue base 40.47%, and epiglottis 11.11% respectively). The rateand of tongue base collapsing was related with Mallampatis(P<0.05) and Friedman stage(P<0.05). The VOTE score was weakly related with Friedman stage(r=0.297, P<0.05), medium related with BMI(r=0.376, P<0.05), AHI(r=0.312, P<0.05) and lowest SpO2(r=0.376, P<0.01). Conclusion:In the VOTE scoring system for DISE, the rate of collapse in tongue base was related with Mallampatis and Friedman stage. The VOTE score was medium related with BMI, AHI and Lowest SpO2, mild related with Friedman stage.


Assuntos
Apneia Obstrutiva do Sono , Endoscopia , Humanos , Faringe , Polissonografia , Sono
18.
Zhonghua Shao Shang Za Zhi ; 35(3): 169-178, 2019 Mar 20.
Artigo em Chinês | MEDLINE | ID: mdl-30897862

RESUMO

Objective: To investigate the effect of human antigen R on lysosomal acidification during autophagy in mouse cardiomyocytes cultured in vitro. Methods: The hearts of 20 C57BL/6 mice aged 1-2 days no matter male or female were isolated to culture primary cardiomyocytes which were used in the following experiments. (1) The cells were divided into 5 groups according to the random number table (the same grouping method below), i. e., normal control group and sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups. The cells in normal control group were routinely cultured for 54.0 h with Dulbecco's modified Eagle medium/nutrient mixture F12 (DMEM/F12) medium (the same regular culture condition below), and the cells in sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups were firstly regularly cultured for 53.5, 53.0, 51.0, 48.0 h and then cultured with replaced sugar-free serum-free medium for 0.5, 1.0, 3.0, and 6.0 h, respectively. The protein expressions of microtubule-associated protein 1 light chain 3 Ⅱ (LC3Ⅱ), autophagy-related protein 5, and adenosine triphosphatase V1 region E1 subunit (ATP6V1E1) were detected by Western blotting. (2) The cells were divided into normal control group and sugar-free serum-free 3.0 h group. The cells in corresponding groups were treated the same as those in experiment (1), and the cell lysosomal acidification level was observed and detected under a laser scanning confocal microscope. (3) Two batches of cells were grouped and treated the same as those in experiment (1). The protein expression of human antigen R in the whole protein of cells of one batch and its protein expression in the cytoplasm and nucleus protein of cells of the other batch were detected by Western blotting. (4) The cells were divided into normal control group, simple control small interfering RNA (siRNA) group, simple human antigen R-siRNA1 (HuR-siRNA1) group, simple HuR-siRNA2 group, sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, sugar-free serum-free+ HuR-siRNA1 group, and sugar-free serum-free+ HuR-siRNA2 group. After 48 hours of regular culture, the cells in simple control siRNA group and sugar-free serum-free+ control siRNA group were transfected with negative control siRNA for 6 h, the cells in simple HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA1 group were transfected with HuR-siRNA1 for 6 h, and the cells in simple HuR-siRNA2 group and sugar-free serum-free+ HuR-siRNA2 group were transfected with HuR-siRNA2 for 6 h. Hereafter, the cells in these 8 groups were continuously cultured for 48 h with regular conditon, and then the cells in normal control group and each simple siRNA-treated group were replaced with DMEM/F12 medium, the cells in the other groups were replaced with sugar-free serum-free medium, and they were cultured for 3 h. The protein expression of human antigen R in the whole protein of cells was detected by Western blotting. (5) Two batches of cells were divided into sugar-free serum-free+ control siRNA group and sugar-free serum-free+ HuR-siRNA1 group, and the cells in corresponding groups were treated the same as those in experiment (4). The distribution and expression of human antigen R in the cells of one batch were observed and detected by immunofluorescence method, and the lysosomal acidification level in the cells of the other batch was observed and detected under a laser scanning confocal microscope. (6) Three batches of cells were divided into sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, sugar-free serum-free+ HuR-siRNA1 group, and sugar-free serum-free+ HuR-siRNA2 group, and the cells in corresponding groups were treated the same as those in experiment (4). The protein expressions of cathepsin D in the whole protein of cells of one batch, human antigen R in the cytoplasm protein of cells of one batch, and ATP6V1E1 in the whole protein of cells of the other batch were detected by Western blotting. (7) The cells were divided into normal control group, sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, and sugar-free serum-free+ HuR-siRNA1 group, and the cells in corresponding groups were treated the same as those in experiment (4). The mRNA expression of ATP6V1E1 in cells was detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction. The sample number of each experiment was 3. Data were processed with independent data t test, one-way analysis of variance, least significant difference t test, and Bonferroni correction. Results: (1) Compared with those of normal control group, the protein expressions of LC3Ⅱ and ATP6V1E1 in the whole protein of cells of sugar-free serum-free 1.0, 3.0, and 6.0 h groups were significantly increased (t=12.16, 4.05, 4.82, 11.64, 3.29, 8.37, P<0.05 or P<0.01). Compared with that of normal control group, the protein expression of autophagy-related protein 5 in the whole protein of cells of sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups was significantly increased (t=6.88, 10.56, 5.76, 9.91, P<0.05 or P<0.01). (2) Compared with 1.03±0.08 of normal control group, the lysosomal acidification level in the cells of sugar-free serum-free 3.0 group (2.92±0.30) was significantly increased (t=6.01, P<0.01). (3) There was no statistically significant difference in the overall comparison of protein expression of human antigen R in the whole protein of cells among the 5 groups (F=1.09, P>0.05). Compared with that of normal control group, the protein expression of human antigen R in the cytoplasm protein of cells was significantly increased in sugar-free serum-free 1.0, 3.0, and 6.0 h groups (t=43.05, 11.07, 5.39, P<0.05 or P<0.01), while the protein expression of human antigen R in the nucleus protein of cells was significantly decreased in sugar-free serum-free 3.0 and 6.0 h groups (t=11.18, 12.71, P<0.01). (4) Compared with that of simple control siRNA group, the protein expression of human antigen R in the whole protein of cells of simple HuR-siRNA1 group and simple HuR-siRNA2 group was significantly decreased (t=4.82, 4.44, P<0.05). Compared with that of sugar-free serum-free+ control siRNA group, the protein expression of human antigen R in the whole protein of cells of sugar-free serum-free+ HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA2 group was significantly decreased (t=4.39, 6.27, P<0.05). (5) Compared with those of sugar-free serum-free+ control siRNA group, the distribution of human antigen R in the cytoplasm of cells and its expression level were significantly decreased in sugar-free serum-free+ HuR-siRNA1 group (t=10.13, P<0.01). Compared with 1.00±0.06 of sugar-free serum-free+ control siRNA group, the lysosomal acidification level (0.73±0.06) in the cells of sugar-free serum-free+ HuR-siRNA1 group was significantly decreased (t=3.28, P<0.01). (6) Compared with those of sugar-free serum-free+ control siRNA group, the protein expressions of cathepsin D in the whole protein of cells, human antigen R in the cytoplasm protein of cells, and ATP6V1E1 in the whole protein of cells were significantly decreased in sugar-free serum-free+ HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA2 group (t=4.16, 3.99, 4.81, 5.07, 11.68, 12.97, P<0.05 or P<0.01). (7) Compared with that of normal control group, the mRNA expression of ATP6V1E1 in the cells of sugar-free serum-free 3.0 h group was significantly increased (t=5.51, P<0.05). Compared with that of sugar-free serum-free+ control siRNA group, the mRNA expression of ATP6V1E1 in the cells of sugar-free serum-free+ HuR-siRNA1 group was significantly decreased (t=5.97, P<0.05). Conclusions: After sugar-free serum-free treatment in vitro, the autophagy in mouse primary cardiomyocytes is activated, the lysosomal acidification is enhanced, and the expression of human antigen R in cytoplasm is increased. Human antigen R function is activated and involved in maintaining lysosomal acidification during autophagy in mouse cardiomyocytes.


Assuntos
Autofagia , Miócitos Cardíacos/metabolismo , Receptores de Antígenos/metabolismo , Animais , Western Blotting , Células Cultivadas , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL
19.
Zhonghua Shao Shang Za Zhi ; 33(5): 295-300, 2017 May 20.
Artigo em Chinês | MEDLINE | ID: mdl-28651421

RESUMO

Objective: To explore the effects of change of activity of vacuolar adenosine triphosphatase (V-ATPase) of myocardial lysosome on myocardial damage in rats after severe burn and its mechanism. Methods: The myocardial lysosomes were extracted from the hearts of 12 SD rats with ultra-high speed gradient density centrifugation, then Western blotting and transmission electron microscope observation were conducted for identification. One hundred and twenty rats were divided into pure burn group, ATP group, normal control group, and bafilomycin group according to the random number table, with 30 rats in each group. Rats in pure burn group and ATP group were inflicted with 40% TBSA full-thickness scald on the back. Immediately after injury, rats in pure burn group were intraperitoneally injected with lactated Ringer's solution in 4 mL·%TBSA(-1)·kg(-1,) and rats in ATP group were intraperitoneally injected with ATP in 0.4 mg/kg at 12 h before burn, immediately after burn, and 12 h after burn. Rats in normal control group did not receive any treatment, and rats in bafilomycin group were intraperitoneally injected with bafilomycin A1 in 0.3 mg/kg at the same time points as those of ATP group. At 24 h after burn, 30 rats from each group were collected for determining activity of V-ATPase of myocardial lysosome with coupled-enzyme assay and the expression of myocardium autophagy-related proteins microtubule-associated protein 1 light chain 3 (LC3) and P62 by Western blotting. Left ventricular arterial blood was collected to detect the content of 5 items of myocardial enzyme spectrum and cardiac troponin T (cTnT). Data were processed with one-way analysis of variance and t test. Results: (1) After identification, both the expression level of lysosome-related membrane protein 1 and purity of lysosome in the sample were high, and the structure of lysosome was intact. (2) At 24 h after burn, the activity values of V-ATPase of myocardial lysosome in rats of pure burn group, ATP group, normal control group, and bafilomycin group were (2.03±0.67), (3.01±0.58), (4.29±0.26), and (1.83±0.52) µmol·mg(-1)·h(-1,) respectively. The activity value of V-ATPase of myocardial lysosome in rats of pure burn group was significantly lower than the values in ATP group and normal control group (with t values respectively 3.14 and 8.87, P values below 0.01). The activity values of V-ATPase of rats in normal control group were significantly higher than those in bafilomycin group (t=11.87, P<0.01). At 24 h after burn, the expressions of myocardial LC3 and P62 in pure burn group were significantly higher than those in ATP group and normal control group (with t values from 3.73 to 5.88, P values below 0.01). The expressions of myocardial LC3 and P62 in normal control group were significantly lower than those in bafilomycin group (with t values respectively 2.64 and 3.07, P<0.05 or P<0.01). At 24 h after burn, the content of 5 items of myocardial enzyme spectrum and cTnT in pure burn group was significantly higher than that in ATP group and normal control group (with t values from 3.24 to 16.72, P values below 0.01). The content of 5 items of myocardial enzyme spectrum and cTnT in normal control group was significantly lower than that in bafilomycin group (with t values from 2.39 to 10. 70, P values below 0.01). Conclusions: The activity of V-ATPase of myocardial lysosome decreased in rats after severe burn, which can result in myocardial damage by inhibiting myocardial autophagy flux.


Assuntos
Adenosina Trifosfatases , Queimaduras/metabolismo , Lisossomos , Miocárdio/metabolismo , Animais , Western Blotting , Queimaduras/patologia , Coração , Miocárdio/patologia , Ratos , Ratos Sprague-Dawley , Lesões dos Tecidos Moles , Índices de Gravidade do Trauma
20.
Zhonghua Shao Shang Za Zhi ; 32(12): 744-751, 2016 Dec 20.
Artigo em Chinês | MEDLINE | ID: mdl-28043299

RESUMO

Objective: To investigate the mechanism of protective effects of tumor necrosis factor receptor associated protein 1 (TRAP1) on hypoxic cardiomyocytes of rats. Methods: Primary cultured cardiomyocytes were obtained from neonatal Sprague-Dawley rats (aged 1 to 3 days) and then used in the following experiments. (1) Cells were divided into group TRAP1 and control group according to the random number table (the same grouping method below), and then the total protein of cells was extracted. Total protein of cells in group TRAP1 was added with mouse anti-rat TRAP1 monoclonal antibody, while that in control group was added with the same type of IgG from mouse. Co-immunoprecipitation and protein mass spectrography analysis were used to determine the possible proteins interacted with TRAP1. (2) Cells were divided into normoxia blank control group (NBC), normoxia+ TRAP1 interference control group (NTIC), normoxia+ TRAP1 interference group (NTI), normoxia+ TRAP1 over-expression control group (NTOC), and normoxia+ TRAP1 over-expression group (NTO), with 1 well in each group. Cells in group NBC were routinely cultured, while cells in the latter four groups were respectively added with TRAP1 RNA interference empty virus vector, TRAP1 RNA interference adenovirus vector, TRAP1 over-expression empty virus vector, and TRAP1 over-expression adenovirus vector. Another batch of cells were divided into group NBC, hypoxic blank control group (HBC), hypoxic+ TRAP1 interference control group (HTIC), hypoxic+ TRAP1 interference group (HTI), hypoxic+ TRAP1 over-expression control group (HTOC), and hypoxic+ TRAP1 over-expression group (HTO), with 1 well in each group. Cells in hypoxic groups were under hypoxic condition for 6 hours after being treated as those in the corresponding normoxia groups, respectively. The mRNA expression of cytochrome c oxidase subunit Ⅱ (COXⅡ) of cells in each group was detected by real time fluorescent quantitive reverse transcription polymerase chain reaction. Experiments were repeated for three times. (3) Cells were divided into group NBC, group HBC, group HTOC, group HTO, hypoxic+ TRAP1 over-expression+ COXⅡinterference control group (HTOCIC), and hypoxic+ TRAP1 over-expression+ COXⅡinterference group (HTOCI), with 3 wells in each group. Cells in the previous 4 groups were treated as those in experiment (2). Cells in group HTOCIC and HTOCI were respectively transfected with COXⅡ RNA interference empty virus vector and COXⅡ RNA interference adenovirus vector, and then both added with TRAP1 over-expression adenovirus vector. The proliferation activity of cells was determined by cell counting kit 8 and microplate reader, and the ratio of death cells was measured by propidium lodide and Hoechst 33342 staining. Another batch of cells were divided into group NBC, group HBC, group HTIC, group HTI, hypoxic+ TRAP1 interference+ COXⅡover-expression control group (HTICOC), and hypoxic+ TRAP1 interference+ COXⅡ over-expression group (HTICO), with 3 wells in each group. Cells in the previous 4 groups were treated as those in experiment (2). Cells in group HTICOC and HTICO were both transfected with TRAP1 RNA interference adenovirus vector, and then respectively added with COXⅡ over-expression empty virus vector and COXⅡ over-expression adenovirus vector. The proliferation activity of cells and the ratio of death cells were detected as before. Experiments were repeated for three times. Data were processed with one-way analysis of variance and LSD test. Results: (1) The expression of TRAP1 was found in cells of group TRAP1, while that was not found in cells of control group. The possible proteins interacted with TRAP1 were keratin, COXⅡ, and an unknown protein with predicted molecular weight 13×103. (2) Compared with that in group NBC, the mRNA expression of COXⅡof cells had no significant change in group NTIC and group NTOC (with P values above 0.05), but significantly decreased in group NTI (P<0.01), and significantly increased in group NTO (P<0.01). Compared with that in group NBC, the mRNA expression of COXⅡof cells in group HBC was significantly decreased (P<0.01). Compared with that in group HBC, the mRNA expression of COXⅡof cells had no significant change in group HTIC and group HTOC (with P values above 0.05), but significantly decreased in group HTI (P<0.01), and significantly increased in group HTO (P<0.01). (3) The proliferation activity of cells in group NBC, group HBC, group HTOC, group HTO, group HTOCIC, and group HTOCI was respectively 0.498±0.022, 0.303±0.018, 0.313±0.032, 0.456±0.031, 0.448±0.034, and 0.335±0.026, and the ratios of death cells in above groups were respectively (4.7±1.5)%, (24.7±3.1)%, (26.0±2.7)%, (13.3±2.5)%, (12.7±2.1)%, and (21.0±1.7)%. Compared with those in group NBC, the proliferation activity of cells in HBC was decreased, while the ratio of death cells was increased (with P values below 0.01). Compared with those in group HBC, the proliferation activity of cells and the ratio of death cells in group HTOC had no significant change (with P values above 0.05), while the proliferation activity of cells was increased and the ratio of death cells was decreased in group HTO (with P values below 0.01). Compared with those in group HTO, the proliferation activity of cells and the ratio of death cells in group HTOCIC had no significant change (with P values above 0.05), while the proliferation activity of cells was decreased and the ratio of death cells was increased in group HTOCI (with P values below 0.01). (4) The proliferation activity of cells in group NBC, group HBC, group HTIC, group HTI, group HTICOC, and group HTICO was respectively 0.444±0.025, 0.275±0.016, 0.283±0.021, 0.150±0.009, 0.135±0.011, and 0.237±0.017, and the ratios of death cells in above groups were respectively (3.7±0.6)%, (21.0±2.7)%, (20.3±3.1)%, (31.7±2.5)%, (33.3±3.2)%, and (19.3±1.5)%. Compared with those in group HBC, the proliferation activity of cells and the ratio of death cells in group HTIC had no significant change (with P values above 0.05). Compared with those in group HBC and group HTIC, the proliferation activity of cells was decreased and the ratio of death cells was significantly increased in group HTI (with P values below 0.01). Compared with those in group HTI, the proliferation activity of cells and the ratio of death cells in group HTICOC had no significant change (with P values above 0.05), while the proliferation activity of cells was increased and the ratio of death cells was significantly decreased in group HTICO (with P values below 0.01). Conclusions: TRAP1 can up-regulate the expression of COXⅡ mRNA, and COXⅡ is one of the downstream effector molecules that TRAP1 mediates its protective effects on hypoxic cardiomyocytes.


Assuntos
Adenoviridae/genética , Proteínas de Choque Térmico HSP90/metabolismo , Miócitos Cardíacos/metabolismo , RNA Mensageiro , Animais , Vetores Genéticos , Proteínas de Choque Térmico HSP90/genética , Camundongos , Miócitos Cardíacos/citologia , Ratos , Ratos Sprague-Dawley , Transfecção , Regulação para Cima
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