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BACKGROUND: Endothelial dysfunction may play a key role in non-obstructive coronary artery atherosclerosis. Our study aimed to evaluate the vascular endothelial function and its influencing factors in patients with non-obstructive coronary artery atherosclerosis. METHODS: A total of 131 consecutive patients with non-obstructive coronary artery atherosclerosis were enrolled. Flow-mediated dilatation (FMD) was measured at baseline and 1-year follow-up. Endothelial progenitor cells (EPCs) were counted by staining the fasting venous blood with antibodies against CD34 and vascular endothelial growth factor receptor 2. RESULTS: Systolic blood pressure, pulse pressure and the levels of HbA1c in participants with baseline FMD < 6% (n = 65) were significantly higher than those with baseline FMD ≥ 6% (n = 66). Baseline FMD was negatively associated with EPC counts (r = - 0.199, P < 0.05) and systolic blood pressure (r = - 0.315, P < 0.01). The 1-year FMD was significantly increased compared to the baseline FMD [(9.31 ± 5.62) % vs (7.31 ± 5.26) %, P < 0.001]. Independent predictors of FMD improvement included elevated EPC counts (OR = 1.104, 95% CI: 1.047-1.165, P < 0.001) and decreased levels of serum creatinine (OR = 0.915, 95% CI: 0.843-0.993, P = 0.034). CONCLUSIONS: Family history of premature cardiovascular diseases, hypertension, elevated systolic pressure, and HbA1c > 6.5% are independent risk factors for endothelial dysfunction in non-obstructive atherosclerotic patients. Elevated peripheral blood EPC counts and decreased levels of serum creatinine are independent predictors of endothelial function improvement.
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Artéria Braquial/fisiopatologia , Doença da Artéria Coronariana/fisiopatologia , Endotélio Vascular/fisiopatologia , Vasodilatação , Idoso , Antígenos CD34/sangue , Biomarcadores/sangue , Pressão Sanguínea , Artéria Braquial/diagnóstico por imagem , Artéria Braquial/metabolismo , Doença da Artéria Coronariana/sangue , Doença da Artéria Coronariana/diagnóstico por imagem , Creatinina/sangue , Diabetes Mellitus/sangue , Células Progenitoras Endoteliais/metabolismo , Endotélio Vascular/metabolismo , Feminino , Hemoglobinas Glicadas/metabolismo , Humanos , Hipertensão/fisiopatologia , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/sangueRESUMO
CONTEXT: Curcumin is a polyphenolic compound extracted from rhizomes of the tropical plant Curcuma longa L. (Zingiberaceae) and it has antitumor, antioxidative, and anti-inflammatory effects. However, its effects on leukemia cell proliferation and invasion are not clear. OBJECTIVE: This study investigates the effects of curcumin on acute monocytic leukemia SHI-1 cells at the molecular level. MATERIALS AND METHODS: The effects of SHI-1 cells treated with 6.25-25 µM curcumin for 12-48 h were measured by MTT assay, flow cytometry, and Matrigel transwell assay; the underlying molecular mechanisms were assessed by quantitative PCR, Western blotting, and gelatin zymography. RESULTS: Treatment of SHI-1 cells with curcumin inhibited cell proliferation in a dose- and time-dependent manner, and the IC50 values at 12, 24, and 48 h were 32.40, 14.13, and 9.67 µM. Curcumin inhibited SHI-1 cell proliferation by arresting the cells in the S-phase, increasing the number of Annexin V-FITC(+)/PI(-) cells and promoting the loss of â³Ψm. The results of PCR and Western blotting showed that curcumin increased the FasL mRNA level; inhibited Bcl-2, NF-κB, and ERK expression; and activated P38 MAPK, JNK, and caspase-3. Additionally, curcumin partially suppressed SHI-1 cell invasion and attenuated the mRNA transcription and secretion of MMP-2 and MMP-9. DISCUSSION AND CONCLUSION: This study demonstrates that curcumin not only induces SHI-1 cell apoptosis, possibly via both intrinsic and extrinsic pathways triggered by JNK, P38 MAPK and ERK signaling, but also partially suppresses SHI-1 cell invasion, likely by reducing the levels of transcription and secretion of MMP-2 and MMP-9.
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Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Curcumina/farmacologia , Leucemia Monocítica Aguda/tratamento farmacológico , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Transdução de Sinais/efeitos dos fármacos , Proteínas Reguladoras de Apoptose/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Regulação Enzimológica da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Concentração Inibidora 50 , Leucemia Monocítica Aguda/enzimologia , Leucemia Monocítica Aguda/genética , Leucemia Monocítica Aguda/patologia , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 9 da Matriz/genética , Potencial da Membrana Mitocondrial/efeitos dos fármacos , NF-kappa B/metabolismo , Invasividade Neoplásica , Pontos de Checagem da Fase S do Ciclo Celular/efeitos dos fármacos , Fatores de TempoRESUMO
Leupaxin (LPXN) is a new member of the Paxillin superfamily, mainly located in focal adhesion plaques, involved in the transduction of multiple signaling pathways, and regulating the proliferation, adhesion and migration of tumor cells. In prostate cancer cells, LPXN is not only involved in the integrin signaling transduction pathway, regulating the proliferation, adhesion and migration of prostate cancer cells, but is also a new androgen receptor (AR) coactivator, regulating the transcription of nuclear AR effect genes, participating in AR signal transduction, and regulating the differentiation and invasion of prostate cancer cells. This review focuses on the molecular structure, special roles and molecular mechanisms of LPXN involved in prostatic carcinoma metastasis.
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Moléculas de Adesão Celular/genética , Fosfoproteínas/genética , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , Moléculas de Adesão Celular/metabolismo , Humanos , Masculino , Metástase Neoplásica , Fosfoproteínas/metabolismo , Neoplasias da Próstata/metabolismo , Receptores Androgênicos/metabolismo , Transdução de SinaisRESUMO
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a new type of coronavirus that has caused fatal infectious diseases and global spread. This novel coronavirus attacks target cells through the interaction of spike protein and angiotensin-converting enzyme II (ACE2), leading to different clinical symptoms. However, for a successful pregnancy, a well-established in-uterine environment includes a specific immune environment, and multi-interactions between specific cell types are prerequisites. The immune-related changes in patients infected with novel coronavirus could interfere with the immune microenvironment in the uterus, leading to fetal loss. We first reviewed the intrauterine environment in the normal development process and the possible pregnancy outcome in the infection state. Then, we summarized the immune response induced by SARS-CoV-2 in patients and analyzed the changes in ACE2 expression in the female reproductive system. Finally, the present observational evidence of infection in pregnant women was also reviewed.
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COVID-19 , Humanos , Feminino , Gravidez , SARS-CoV-2/metabolismo , Enzima de Conversão de Angiotensina 2 , Peptidil Dipeptidase A/metabolismo , Resultado da GravidezRESUMO
This study aimed to determine the values of the half-effective dose (ED50) and 95% effective dose (ED95) of remimazolam besylate used in the procedural sedation of endoscopic retrograde cholangiopancreatography (ERCP). Sixty patients who fulfilled the inclusion and exclusion criteria of this study were selected. Sufentanil was administered intravenously and remimazolam besylate was administered 2 min later. ERCP treatment was feasible when the modified alertness/sedation (MOAA/S) score was ≤2. If choking or movement occurred during duodenoscope placement, it was considered as a positive reaction. The dose was increased in the next patient; otherwise, it was considered as a negative reaction, and the dose was reduced in the next patient. The ED50 and ED95 values and 95% confidence interval (CI) of remimazolam besylate were calculated by Probit regression analysis. All 60 patients completed the trial. The ED50 and ED95 values of remimazolam besylate were 0.196 and 0.239 mg/kg, respectively, for the procedural sedation of ERCP. The time of MOAA/S score ≤ 2 was (82.58 ± 21.70) s, and the mean time of awakening was (9.03 ± 5.64) min. Transient hypotension was observed in two patients without medical intervention. The ED50 and ED95 values of remimazolam besylate used in the procedural sedation of ERCP were 0.196 and 0.239 mg/kg, and the dose of the medications has definite efficacy and good safety.
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OBJECTIVE: To investigate the effects of Tongmai Yangxin Pill (TMYXP) combined with metoprolol tartrate or metoprolol alone for the treatment of premature ventricular complex (PVC) in patients with symptomatic frequent PVC. METHODS: A total of 584 patients with symptomatic frequent PVC were randomly assigned (in a 1:1 ratio) into two groups: study group [n = 292, TMYXP (40 pills twice/day, orally) combined with metoprolol tartrate (25 mg twice/day, orally)] and control group [n = 292, metoprolol tartrate (25 mg twice/day, orally) plus placebo pill (40 pills twice/day, orally)]. The total treatment period was eight weeks. RESULTS: After eight weeks of treatment, the total effective rate of reduction of PVC in the study group and the control group were 76.4% and 51.4%, respectively (P < 0.001). TMYXP combined with metoprolol tartrate demonstrated a signiï¬cantly greater reduction of the frequency of PVCs compared with the metoprolol tartrate alone (-4537 times/24 h vs. -3013 times/24 h, P < 0.001). The study group also showed a better result compared with the control group with respect to PVC related symptoms. In terms of New York Heart Association classification improvement, the total effective rates were 21.9% in the study group and 12.4% in the control group ( P < 0.05). Both the study group and the control group exhibited improvements in echocardiographic indexes. Left ventricular ejection fraction was significantly improved in the study group compared with the control group ( P < 0.05). There was no significant difference in the incidence of adverse events between the two groups. CONCLUSIONS: Compared with metoprolol tartrate alone, TMYXP combined with metoprolol tartrate could more effectively reduce the frequency of PVC and alleviated PVC related symptoms, and improve cardiac function in patients with symptomatic PVC.
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OBJECTIVE: To evaluate the effect of songlingxuemaikang (SLXMK) on mild essential hypertension in patients in terms of endothelial function. METHODS: We enrolled 90 patients with mild essential hypertension in Xuanwu Hospital from January 2016 to December 2016. To evaluate the effects of SLXMK, the 90 patients were randomly assigned at a 2â¶1 ratio into 2 groups: the SLXMK group (500 mg per capsule, 4500 mg/d, n = 60) and the losartan potassium group (50 mg per table, 50 mg/d, n = 30). The total study period was 12 weeks, and the changes of blood pressure, laboratory test and endothelium function were compared between two groups. RESULTS: After 12 weeks of treatment with SLXMK, blood pressure (BP) and plasma lipid levels significantly improved (P < 0.05). Meanwhile, the reactive hyperemia index (RHI) increased in the SLXMK group (P < 0.05). The results of multivariate logistic regression analyses examining the association of selected variables with showed that high level of oxidized low density lipoprotein (ox-LDL) was positively associated with endothelial dysfunction. CONCLUSION: SLXMK not only effectively decreased BP and plasma lipid levels, but also reduced ox-LDL and RHI in patients with mild essential hypertension. And SLXMK might improve endothelial function through decreasing the circulating ox-LDL.
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Hipertensão , Pressão Sanguínea , Hipertensão Essencial/tratamento farmacológico , Humanos , Hipertensão/tratamento farmacológicoRESUMO
BACKGROUND: Endothelial dysfunction is the initial stage in atherosclerotic formation and progression and is associated with high serum uric acid (SUA) level. We hypothesized that reactive hyperemia index (RHI), which reflects endothelial function, is associated with SUA levels in elderly individuals with untreated mild hypertension. METHODS: We recruited 123 patients ≥ 60 years with untreated mild hypertension. The association between SUA level and RHI was analyzed using univariate correlation analysis and multiple regression analysis. The receiver operating characteristic (ROC) curve was performed to validate the cutoff value of SUA that can be used to predict endothelial dysfunction. RESULTS: The serum uric acid level significantly increased in the RHI < 1.67 group, and this result was still observed in the subgroup of men. RHI was inversely associated with SUA level (P = 0.006) and the association was still observed after adjusting for factors, such as age, sex, smoking status, and creatinine level (P = 0.014). In the subgroup analysis, a positive association was observed only in men. In the ROC curve analysis, the optimal cutoff values of SUA for predicting endothelial dysfunction was 293.5 µmol/L in elderly mild hypertension patients and 287.0 µmol/L in men. CONCLUSION: A high SUA level was considered an independent predictor of endothelial dysfunction among elderly individuals, particularly men with untreated mild hypertension.
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The aim of the present study was to investigate the effects of decreasing leupaxin (LPXN) expression on the proliferation and invasion of human acute monocytic leukemia SHI-1 cells. The transfection efficiency of fluorescein amidite (FAM)-small interfering RNA (siRNA) was determined using flow cytometry, and the protein expression levels of LPXN, phosphorylated (p)-c-Jun N-terminal kinase (JNK), p-p38 mitogen-activated protein kinase (p38 MAPK) and p-extracellular-signal-regulated kinase (ERK) were detected by western blot analysis. Proliferation was determined using the cell counting kit-8 reagent and cellular transmembrane invasion ability was determined using a Transwell chamber system. The gelatinase levels of matrix metalloproteinase (MMP)-2 and MMP-9 in the cell culture supernatant were also analyzed by gelatin zymography. In SHI-1 cells, the optimal transfection conditions of siRNA were a cell density of 4×105 cells/ml and a ratio of siRNA/Lipofectamine® 2000 of 200 pmol/1 µl. The highest transfection efficiency of FAM-siRNA was 74.5%. In the present study, L2-siRNA was selected to effectively decrease the expression of LPXN. Following downregulation of LPXN expression by L2-siRNA, proliferation inhibition rates increased to 27.043±2.051 and cell transmembrane invasion rates decreased to 25.270±2.145 (P<0.05). The results of the western blot analysis and the gelatin zymography indicated that downregulation of LPXN expression increased the expression of p-p38 MAPK and p-JNK, and attenuated the secretion levels of MMP-2 and MMP-9. However, downregulation of LPXN expression had no effect on p-ERK expression in SHI-1 cells. The results of the present study indicated that downregulation of LPXN expression decreased the malignant proliferation and transmembrane invasion of SHI-1 cells by activating JNK and p38 MAPK, and inhibiting MMP-2 and MMP-9 secretion.
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BACKGROUND: Elevated serum phosphorus levels may be associated with adverse outcomes in cardiovascular disease. This study aimed to investigate the relation between serum phosphorus levels and risk of all-cause mortality in Chinese patients with ST-segment elevation myocardial infarction (STEMI) who had preserved renal function at baseline. METHODS: We enrolled patients with STEMI who had preserved renal function at baseline in Xuanwu Hospital from January 2011 to December 2016. Those patients were divided into four groups based on serum phosphorus levels. All-cause mortality rates were compared between groups. Mean duration of follow up was 54.6 months. We used Cox proportional-hazards models to examine the relation between serum phosphorus levels and all-cause mortality after adjustment for potential confounders. RESULTS: 1989 patients were involved and 211 patients (10.6%) died during follow-up. Based on serum phosphorus levels, patients were categorized into the following groups: < 2.50 mg/dL (n = 89), 2.51-3.50 mg/dL (n = 1066), 3.51-4.50 mg/dL (n = 672) and > 4.50 mg/dL (n = 162), respectively. The lowest mortality occurred in patients with serum phosphorus levels between 2.51-3.50 mg/dL, with a multivariable-adjusted hazard ratio of 1.19 (95% CI: 0.64-1.54), 1.37 (95% CI: 1.22-1.74), and 1.46 (95% CI: 1.35-1.83) in patients with serum phosphorus levels of < 2.50 mg/dL, 3.51-4.50 mg/dL and > 4.50 mg/dL, respectively. CONCLUSIONS: Elevated serum phosphorus levels were associated with all-cause mortality in Chinese patients with STEMI who had preserved renal function at baseline.
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BACKGROUND: Reserpine is currently used by millions of Chinese hypertensive patients, in spite of the continued concern of its depressogenic effect, even when used in low dose. This study aimed to investigate the association between low-dose reserpine use and depression in older Chinese hypertensive patient. METHODS: In this cross-sectional, case-control study, we recruited patient aged 60 years or over who had regularly taken one or two tables of "compound reserpine and triamterene tablets (CRTTs)" for more than one year (reserpine user) from 26 community health centers located in 10 provinces in China. For each patient who took CRTTs, we selected an age (within five years) and sex matched hypertensive patient who had never taken any drugs containing reserpine (non-reserpine user) as control. Depressive symptoms were evaluated using a Chinese depression scale adapted from the Zung Self-Rating Depression Scale. Demographic, clinical data and laboratory examination results within six months were collected. RESULTS: From August 2018 to December 2018, 787 reserpine user and 787 non-reserpine user were recruited. The mean age of all study subjects was 70.3 years, with about equal numbers of males and females. The mean depression score was 40.4 in reserpine users and 40.6 in non-reserpine users (P = 0.7). The majority of study subject had a depression score < 53 (87.6% in reserpine users and 88.2% in non-reserpine users, respectively). There were no significant differences in the prevalence of mild, moderate or severe depression in reserpine users and non-reserpine users. CONCLUSIONS: There is no association between low-dose reserpine use and depression in older hypertensive patient. The role of reserpine in the treatment and control of hypertension should be reconsidered; and further studies, especially randomized, controlled clinical trials to compare efficacy and safety of reserpine and other widely recommended anti-hypertensive agents are needed.
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OBJECTIVE: To explore the effect of LPXN overexpression on the proliferation, adhesion and invasion of THP-1 cells and its possible mechanism. METHODS: A THP-1 cell line with stable overexpression of LPXN was constucted by using a lentivirus method, CCK-8 was used to detect the proliferation of cells, adhesion test was used to evaluate adhesion ablity of cells to Fn. Transwell assay was used to detect the change of invasion capability. Western blot was used to detect expression of LPXN, ERK, pERK and integrin α4, α5, ß1, the Gelatin zymography was applied to detect activity of MMP2/MMP9 secreted by the THP-1 cells. RESULTS: Successful establishment of THP-1 cells with LPXN overexpression (THP-1 LPXN) was confirmed with Western blot. THP-1 LPXN cells were shown to proliferate faster than the control THP-1 vector cells. Adhesion to Fn and expression of ERK, integrin α4, α5 and ß1 in the THP-1 LPXN cells were higher than that in the control cells. Invasion across matrigel and enhanced activity of MMP2 could be detected both in the THP-1 LPXN cells as compared with the control cells. CONCLUSION: Ectopically ovexpression of LPXN may promote proliferation of THP-1 cells through up-regulation of ERK; promote adhesion of THP-1 cells through up-regulating the integrin α4/ß1 as well as integrin α5/ß1 complex; promote invasion of THP-1 cells through activating MMP2.
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Moléculas de Adesão Celular/metabolismo , Adesão Celular , Proliferação de Células , Fosfoproteínas/metabolismo , Células THP-1 , Linhagem Celular Tumoral , Humanos , Metaloproteinase 2 da Matriz , Metaloproteinase 9 da MatrizRESUMO
A novel dissolved carbon dioxide flotation after emulsification microextraction (DCF-EME) method was proposed for the determination of four triazole pesticides in water samples coupled with gas chromatography-mass spectrometry (GC-MS) in a home-made glass round flask. The DCF-EME method is based on a rapid and simple phase separation of low-density organic solvent (toluene) from the aqueous phase via introducing a saturated NaHCO3 solution into the acidified sample (0.1 mol L-1 HCl); then analytes were extracted in toluene. Various parameters affecting the extraction process were optimized. Under the optimal conditions, the recoveries for four pesticides ranged from 82.8 to 121.2%. Meanwhile the limits of detection were at the range of 0.14 - 1.04 µg L-1, and the preconcentration factors were varied between 342 and 473 for different triazoles. The method is simple, fast and environmentally friendly, being successfully applied for the determination of triazole pesticides in water samples.
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Dióxido de Carbono/química , Cromatografia Gasosa-Espectrometria de Massas/métodos , Microextração em Fase Líquida/métodos , Resíduos de Praguicidas/análise , Triazóis/análise , Poluentes Químicos da Água/análise , Emulsões , Modelos Teóricos , SolubilidadeRESUMO
This study was purposed to explore the effects induced by puerariae radix flavones (PRF) on human acute myeloid leukemia SHI-1 cells, apoptosis induced by PRF in vitro and its molecular mechanism. SHI-1 cells were treated with PRF in various concentration, then the inhibitory effect of cell proliferation were detected by MTT method, the cell cycle was analyzed by flow cytometry, the mRNA expression levels of Caspase-3, Caspase-8, Caspase-9, Bcl-2 and MLL-AF6 were detected by real-time polymerase chain reaction (R-T PCR), the protein expression levels of MAPK, p-MAPK and NF-κB were assayed by Western blot, and the activity of MMP was analyzed by Gelatin zymography. The results indicated that the PRF could inhibit the proliferation of SHI-1 cells in a time-and dose-dependent manner, and the cell cycle was arrested in S phase. When SHI-1 cells were treated with 25, 50 and 75 µg/ml PRF respectively, mRNA levels of Caspase-3, Caspase-8 and Caspase-9 increased in a dose-dependent manner (P < 0.05), Bcl-2 mRNA decreased in a concentration-dependent manner (P > 0.05), and the mRNA level of fusion gene MLL-AF6 did not changed as compared with the control group. Different concentration of PRF was used to treat SHI-1 cells, the expression levels of intracellular JNK, p-JNK, P38 MAPK and p-P38 MAPK increased in the concentration-dependent manner (P < 0.01); the expression of p-ERK1/2 and NF-κB decreased in the concentration-dependent manner, and the activity of MMP-2 and MMP-9 in the cell supernatant did not change in each groups. It is concluded that a certain concentration of PRF can induce the apoptosis of SHI-1 cells in vitro, its molecular mechanism may be related to the activation of Caspase hydrolase, activation of MAPK, downregulation of NF-κB, Bcl-2 and other signal molecules. However, it seemed that all these effects are not relate with the MLL-AF6 fusion gene.
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Apoptose/efeitos dos fármacos , Flavonas/farmacologia , Pueraria , Linhagem Celular Tumoral , Humanos , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Proteína de Leucina Linfoide-Mieloide/metabolismo , NF-kappa B/metabolismo , Proteínas de Fusão Oncogênica/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Pueraria/química , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
One hundred and fifty one soil samples from 125 sampling points were collected near E-waste dismantling sites and analyzed for polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (PCBs). Total concentration of PCDD/Fs in topsoil samples ranged from 280 pg x g(-1) to 7 010 pg x g(-1) dry weight with the mean value of 1 380 pg x g(-1) dry weight. The mean value of the second layer and the deepest layer is 63% and 38% of that of top soils, respectively. The toxic equivalent quantity (TEQ) of PCDD/Fs in top soils ranged from 1.4 pg x g(-1) to 94.8 pg x g(-1) (I-TEQ). According to the guidelines for TEQ concentrations established in Germany, 19 of 125 topsoil samples (15% of the total topsoil samples analyzed) could not be expected to pose human health hazards. The rest of 85% of top soils needs investigations of source identification. If the contribution of PCB to TEQ was considered, 98% of top soils need investigations of source identification. Principle component analysis (PCA) is used to identify the PCDD/Fs sources. The homologue profiles of PCDD/Fs showed that dismantling activities are the main emission resources of the PCDD/Fs from thermal processes, also are the main emission resources of the PCDD/Fs in soils of this area.
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Benzofuranos/análise , Dioxinas/análise , Resíduo Eletrônico , Bifenilos Policlorados/análise , Dibenzodioxinas Policloradas/análogos & derivados , Poluentes do Solo/análise , Dibenzofuranos Policlorados , Dibenzodioxinas Policloradas/análise , Solo/químicaRESUMO
AIM: To investigate the effects of the WWOX gene on the human hepatic carcinoma cell line SMMC-7721. METHODS: Full-length WWOX cDNA was amplified from normal human liver tissues. Full-length cDNA was subcloned into pEGFP-N1, a eukaryotic expression vector. After introduction of the WWOX gene into cancer cells using liposomes, the WWOX protein level in the cells was detected through Western blotting. Cell growth rates were assessed by methyl thiazolyl tetrazolium (MTT) and colony formation assays. Cell cycle progression and cell apoptosis were measured by flow cytometry. The phosphorylated protein kinase B (AKT) and activated fragments of caspase-9 and caspase-3 were examined by Western blotting analysis. RESULTS: WWOX significantly inhibited cell proliferation, as evaluated by the MTT and colony formation assays. Cells transfected with WWOX showed significantly higher apoptosis ratios when compared with cells transfected with a mock plasmid, and overexpression of WWOX delayed cell cycle progression from G1 to S phase, as measured by flow cytometry. An increase in apoptosis was also indicated by a remarkable activation of caspase-9 and caspase-3 and a dephosphorylation of AKT (Thr308 and Ser473) measured with Western blotting analysis. CONCLUSION: Overexpression of WWOX induces apoptosis and inhibits proliferation of the human hepatic carcinoma cell line SMMC-7721.
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Apoptose/genética , Carcinoma Hepatocelular/metabolismo , Proliferação de Células , Oxirredutases/metabolismo , Proteínas Supressoras de Tumor/metabolismo , Carcinoma Hepatocelular/genética , Caspase 1/metabolismo , Caspase 9/metabolismo , Pontos de Checagem do Ciclo Celular , Linhagem Celular Tumoral , Humanos , Oxirredutases/genética , Proteínas Supressoras de Tumor/genética , Oxidorredutase com Domínios WWRESUMO
OBJECTIVE: To compare therapeutic effects of three fixation methods with three Kirschner wires,and to find the best fixation method. METHODS: From July 2008 to May 2009, 60 patients with humeral supracondylar fractures were treated. Among patients in Group A, 11 patients were male and 9 patients were female, ranging from 2 to 13 years old,with an average of (5.4 +/- 0.5)years;all the patients in Group A were fresh closed fractures without nerve and blood vessel injuries, 13 patients were type II and 7 patients were type III according to Gartland classification; and all the patients in Group A were treated by three Kirschner wires fixation with the third Kirschner different wire fixed through radial edge of coronoid fossa. Among patients in Group B, 13 patients were male and 7 patients were female,ranging from 2 to 11 years old, with an average of (6.1 +/- 0.4) years; all the patients in Group B were fresh closed fractures without blood vessel injuries, 11 patients were type II and 9 patients were type III according to Gartland classification, 2 patients had ulnar nerve injuries before treatment; and all the patients in Group B were treated by three Kirschner wires fixation with the third Kirschner different wire fixed through ulnar edge of coronoid fossa. Among patients in Group C, 8 patients were male and 12 patients were female, ranging from 3 to 14 years old, with an average of (7.4 +/- 0.6) years; all the patients in Group C were closed fresh fractures without blood vessel injuries, 7 patients were type II and 13 patients were type III according to Gartland classification, 2 patients had radial nerve injuries before treatment; and all the patients in Group C were treated by three Kirschner wires fixation with the third Kirschner different wire fixed through middle of coronoid fossa. After 1 year of treatment,the Carrying and Baumann angles were measured on the X-ray of all patients in the three groups, and the motion range of elbow joint was observed. The Flynn evaluation criteria were used to compare therapeutic effects among the three groups. RESULTS: All the patients were followed up. The function loss of elbow joint were (14.04 +/- 3.25) degrees in Group A, (13.14 +/- 2.34) degrees in Group B, and (9.25 +/- 2.56) degrees in Group C. Changes of Carrying angle were (7.64 +/- 1.78) degrees in Group A; (7.38 +/- 1.68) degrees in Group B, and (5.27 +/- 1.13) degrees in Group C. Changes of Baumann angle were (28.55 +/- 3.23) degrees in Group A, (27.55 +/- 2.23) degrees in Group B, (21.45 +/- 1.73) degrees in Group C. According to Flynn criteria, there were 11 patients got excellent result, 4 good and 5 poor in Group A; in Group B, 12 patients got excellent result, 3 good and 5 poor; in Group C, 11 patients got excellent result, 6 good and 3 poor. The therapeutic effects of Group C was better than those of Group A and Group B. CONCLUSION: There are different effects among three kinds of operative approaches for the humerus supracondylar fractures. The fixation method with one Kirschner wire through middle part of coronoid fossa is better than the other two fixation methods.