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Steroid-resistant nephrotic syndrome (SRNS) is the second cause of chronic kidney disease in children. The SRNS has high risk of rapid progression to end-stage renal disease. With the advancement of high-throughput sequencing technology, more than 70 monogenic mutation having the Mendelian inheritance patterns are identified to be associated with SRNS. Most of these genes are involved in podocyte function. Accurate diagnosis of monogenic mutation in SRNS patients helps with guiding clinical treatment protocols and genetic counseling, avoiding the excessive use of steroids/immunosuppressive therapy, and opening up possibilities for targeted therapies in SRNS patients. In this article, our research team summarizes and generalizes the molecular mechanisms, genetic testing, and specific treatment for the major types of monogenic mutations associated with SRNS.
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ObjectiveThis study aimed at conducting retrospective analysis of the clinical symptoms and genetic mutations in 20 children with Gitelman syndrome treated at the Affiliated Children′s Hospital of Nanjing Medical University from August 2015 to November 2022 and also explored the molecular mechanism of the pathogenic high-frequency mutation D486N in the Chinese population.MethodsWe collected the clinical manifestations, growth and development status, laboratory examination results, and SLC12A3 gene variations of the patients. We distinguished the wild-type and mutant SLC12A3 genes overexpressed in human embryonic kidney 293T cells (HEK293T). We used protein immunoblotting to detect the expression level of NCC, and used immunofluorescence techniques to examine the subcellular localization of NCC. In addition, we investigated the impact of the high-frequency SLC12A3 gene mutation D486N on NCC protein expression and localization.ResultsIn the 20 patients with Gitelman syndrome, all of them had hypokalemia. We indemnified twenty-six SLC12A3 gene mutations, 13 of which are missense mutation, 1 of which synonymous mutation, 1 nonsense mutation, 4 frameshift mutation, and 7 splicing site mutation. Among them, four mutations (p.T235K, c.1096-1G > A, p.A464A, and c.2660+1_2660+2insT) were novel mutations.ConclusionsWe found the preliminary evidence that the high-frequency mutation D486N in the Chinese population affected the expression of total and membrane-bound NCC protein and influenced the membrane localization of NCC protein. The findings of this study provides experimental evidence for genetic counseling, diagnosis, and treatment of Gitelman syndrome.
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ObjectiveTo analyze the epidemic characteristics of measles and rubella in Pudong New Area of Shanghai from 2013 to 2022, and to provide data support for the elimination of measles and rubella. MethodsEnzyme linked immunosorbent assay was used to detect IgM antibodies in serum samples. The sequence of 630 nucleotides at the C-terminal of N gene of measles virus was amplified by reverse transcription-polymerase chain reaction and the phylogenic tree was constructed. ResultsA total of 1 529 suspected cases of measles were detected from 2013 to 2022, among which the positive rate of measles IgM antibody was 33.55% (513/1 529). The highest positive rate (20.73%) was from March to May , and the positive rate of rubella IgM antibody was 6.80% (104/1 529). The positive rate of both IgM was higher in males than that in females (P<0.05). The IgM against measles was mainly detected in 0‒ years old (63.16%, 96/152) and 20‒ years old (45.61%, 161/353). The IgM against rubella was mainly detected in 10‒20 years old (27.27%, 18/66). The IgM antibody could be detected more easily from 4 to 28 days after eruption, and the IgM antibody positive rate of measles/rubella from 2020 to 2022 was significantly lower than previous years (2013‒2019). There were 2 D8 genotype strains, and the rest were H1a gene subtypes. ConclusionThe positive rate of IgM antibodies against measles/rubella in Pudong New Area of Shanghai decreased significantly. People aged 0‒ years and 20‒ years old are more susceptible to measles, and rubella is concentrated in 10‒ years old. It is necessary to strengthen the vaccination of school-age children, in order to achieve the goal of eliminating measles. The age group with high risk of exposure should be checked for vaccination status to ensure the enhanced immunization, and the surveillance of imported measles cases should be strengthened.
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Steroid-resistant nephrotic syndrome (SRNS) is the second cause of chronic kidney disease in children. The SRNS has high risk of rapid progression to end-stage renal disease. With the advancement of high-throughput sequencing technology, more than 70 monogenic mutation having the Mendelian inheritance patterns are identified to be associated with SRNS. Most of these genes are involved in podocyte function. Accurate diagnosis of monogenic mutation in SRNS patients helps with guiding clinical treatment protocols and genetic counseling, avoiding the excessive use of steroids/immunosuppressive therapy, and opening up possibilities for targeted therapies in SRNS patients. In this article, our research team summarizes and generalizes the molecular mechanisms, genetic testing, and specific treatment for the major types of monogenic mutations associated with SRNS.
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ObjectiveThis study aimed at conducting retrospective analysis of the clinical symptoms and genetic mutations in 20 children with Gitelman syndrome treated at the Affiliated Children′s Hospital of Nanjing Medical University from August 2015 to November 2022 and also explored the molecular mechanism of the pathogenic high-frequency mutation D486N in the Chinese population.MethodsWe collected the clinical manifestations, growth and development status, laboratory examination results, and SLC12A3 gene variations of the patients. We distinguished the wild-type and mutant SLC12A3 genes overexpressed in human embryonic kidney 293T cells (HEK293T). We used protein immunoblotting to detect the expression level of NCC, and used immunofluorescence techniques to examine the subcellular localization of NCC. In addition, we investigated the impact of the high-frequency SLC12A3 gene mutation D486N on NCC protein expression and localization.ResultsIn the 20 patients with Gitelman syndrome, all of them had hypokalemia. We indemnified twenty-six SLC12A3 gene mutations, 13 of which are missense mutation, 1 of which synonymous mutation, 1 nonsense mutation, 4 frameshift mutation, and 7 splicing site mutation. Among them, four mutations (p.T235K, c.1096-1G > A, p.A464A, and c.2660+1_2660+2insT) were novel mutations.ConclusionsWe found the preliminary evidence that the high-frequency mutation D486N in the Chinese population affected the expression of total and membrane-bound NCC protein and influenced the membrane localization of NCC protein. The findings of this study provides experimental evidence for genetic counseling, diagnosis, and treatment of Gitelman syndrome.
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Objective To evaluate the intervention effect of exercise on cancer related cognitive impairment of breast cancer patients,and to provide evidence-based evidence for the choices of rehabilitation programs for cognitive impairment.Methods The PubMed,Embase,Web of Science,Cochrane Library,Joanna Briggs Institute Library(JBI),CINAHL,CNKI,Wanfang,VIP and CBM were searched by computer for randomized controlled trials on the impact of exercise on cancer-related cognitive impairment in patients with breast cancer.The retrieval time limit was from the establishment of the database to October 2022.The quality of the included studies was evaluated according to Cochrane Handbook 5.1.0.RevMan5.4 was used for meta-analysis.Results A total of 18 publications including 1310 patients were included.The results showed that exercise could improve self-reported cancer-related cognitive impairment[SMD=0.33,95%CI(0.21,0.46),P<0.001],executive function[SMD=-0.27,95%CI(-0.42,-0.11),P<0.001]and attention[SMD=-0.37,95%CI(-0.60,-0.14),P<0.01],alleviate cancer-related fatigue[SMD=-0.56,95%CI(-0.79,-0.34),P<0.001],and reduce depression[SMD=-0.73,95%CI(-1.17,-0.30),P<0.001],but it has no significant effect on the memory of patients with breast cancer[SMD=0.05,95%CI(-0.16,0.27),P=0.64].Conclusion Exercise can improve the self-reported cancer-related cognitive impairment,executive function and attention of breast cancer patients,alleviate cancer-related fatigue,reduce depression,but it has no significant effect on improving memory,which still needs further verification.
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Objective:To evaluate the correlation between age and renal outcomes in patients with stage 2-4 chronic kidney disease (CKD) and the impact of age on CKD outcomes in kidney diseases of different etiologies.Methods:A prospective cohort study included 470 patients with stage 2-4 CKD. The Kaplan Meier method was used to analyze the differences in CKD outcomes among different age groups. The independent risk factors for CKD progression were analyzed using a multivariate Cox regression model. We adjusted for baseline differences in risk factors for CKD outcomes between two age groups using propensity score matching (PSM).Results:Among 470 patients, 39 cases of end-stage renal disease (ESRD) events (all starting dialysis) and 51 deaths were observed. The Kaplan Meier survival curve ( P=0.039) and Cox regression univariate survival analysis ( P=0.043) both showed that <60 years old is a risk factor for CKD patients to progress to ESRD. In multivariate Cox regression, age remained an independent risk factor for the progression of CKD patients (hazard ratio 0.386, 95% CI: 0.163-0.916; P=0.031). For kidney diseases with different causes, in patients with hypertensive kidney damage ( P=0.024) and primary glomerulonephritis ( P=0.047), the cumulative incidence rate of ESRD in patients <60 years old was higher than that in patients ≥60 years old. There was no statistically significant difference in all-cause mortality rates between patients aged <60 and ≥60 years old ( P=0.646). Conclusions:Elderly patients with stage 2-4 CKD have a lower ESRD risk than younger patients. This discovery helps nephrologists and decision-makers optimize the management of elderly CKD patients.
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Objective:To investigate the clinical effects of a single lateral-rectus approach (LRA) in the Ti-Robot-assisted treatment of acetabular fractures involving the posterior column.Methods:A retrospective study was conducted to analyze the clinical data of 16 patients with acetabular fracture who had been admitted to Department of Trauma Orthopedics, Taihe Hospital from August 2021 to March 2023. There were 13 males and 3 females with a mean age of 51.5 (35.8, 56.8) years. By the Judet-Letournel classification, there were 4 anterior and posterior semi-transverse fractures, 2 T-shaped fractures, and 10 double-column fractures. The time from injury to operation was 6.0 (5.3, 7.8) d. All their acetabular fractures involving the posterior column were treated by a single LRA with the assistance of a Ti-Robot. Operation time, blood loss, complications, quality of fracture reduction, modified Merle d'Aubigné & Postel scores and acetabular recovery at the last follow-up were recorded.Results:In this cohort, the operation time was (4.1±0.6) h and the blood loss 400 (300, 575) mL. By the Matta evaluation, the fracture reduction was excellent in 10 cases, good in 5 cases, and poor in 1 case. The follow-up time was (11.3±3.9) months. Bone union was achieved in all fractures after 3.0 (2.3, 4.0) months. By the modified Merle d'Aubigné & Postel scores at the last follow-up, 9 cases were excellent, 6 cases were good and 1 case was fair. Injury to the lateral femoral cutaneous nerve occurred in 1 patient, and deep venous thrombosis in 6 patients. Follow-ups revealed no iatrogenic vascular injury, loosening or rupture of internal fixation, or incision infection.Conclusion:A single LRA approach can be used in the minimally invasive treatment assisted by a Ti-Robot for acetabular fractures involving the posterior column, demonstrating advantages of minimal invasion, high reduction quality and good functional recovery of the hip joint.
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Photoplethysmography (PPG) is often affected by interference, which could lead to incorrect judgment of physiological information. Therefore, performing a quality assessment before extracting physiological information is crucial. This paper proposed a new PPG signal quality assessment by fusing multi-class features with multi-scale series information to address the problems of traditional machine learning methods with low accuracy and deep learning methods requiring a large number of samples for training. The multi-class features were extracted to reduce the dependence on the number of samples, and the multi-scale series information was extracted by a multi-scale convolutional neural network and bidirectional long short-term memory to improve the accuracy. The proposed method obtained the highest accuracy of 94.21%. It showed the best performance in all sensitivity, specificity, precision, and F1-score metrics, compared with 6 quality assessment methods on 14 700 samples from 7 experiments. This paper provides a new method for quality assessment in small samples of PPG signals and quality information mining, which is expected to be used for accurate extraction and monitoring of clinical and daily PPG physiological information.
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Fotopletismografía , Aprendizaje Automático , Redes Neurales de la ComputaciónRESUMEN
BACKGROUND: Guizhou is located in the southwest of China with high multidrug-resistant tuberculosis (MDR-TB) epidemic. To fight this disease, Guizhou provincial authorities have made efforts to establish MDR-TB service system and perform the strategies for active case finding since 2014. The expanded case finding starting from 2019 and COVID-19 pandemic may affect the cases distribution. Thus, this study aims to analyze MDR-TB epidemic status from 2014 to 2020 for the first time in Guizhou in order to guide control strategies. METHODS: Data of notified MDR-TB cases were extracted from the National TB Surveillance System correspond to population information for each county of Guizhou from 2014 to 2020. The percentage change was calculated to quantify the change of cases from 2014 to 2020. Time trend and seasonality of case series were analyzed by a seasonal autoregressive integrated moving average (SARIMA) model. Spatial-temporal distribution at county-level was explored by spatial autocorrelation analysis and spatial-temporal scan statistic. RESULTS: Guizhou has 9 prefectures and 88 counties. In this study, 1,666 notified MDR-TB cases were included from 2014-2020. The number of cases increased yearly. Between 2014 and 2019, the percentage increase ranged from 6.7 to 21.0%. From 2019 to 2020, the percentage increase was 62.1%. The seasonal trend illustrated that most cases were observed during the autumn with the trough in February. Only in 2020, a peak admission was observed in June. This may be caused by COVID-19 pandemic restrictions being lifted until May 2020. The spatial-temporal heterogeneity revealed that over the years, most MDR-TB cases stably aggregated over four prefectures in the northwest, covering Bijie, Guiyang, Liupanshui and Zunyi. Three prefectures (Anshun, Tongren and Qiandongnan) only exhibited case clusters in 2020. CONCLUSION: This study identified the upward trend with seasonality and spatial-temporal clusters of MDR-TB cases in Guizhou from 2014 to 2020. The fast rising of cases and different distribution from the past in 2020 were affected by the expanded case finding from 2019 and COVID-19. The results suggest that control efforts should target at high-risk periods and areas by prioritizing resources allocation to increase cases detection capacity and better access to treatment.
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COVID-19 , Tuberculosis Resistente a Múltiples Medicamentos , COVID-19/epidemiología , China/epidemiología , Humanos , Pandemias , Análisis Espacio-Temporal , Tuberculosis Resistente a Múltiples Medicamentos/epidemiologíaRESUMEN
With the aging and lifestyle changes of the residents in arsenicosis area, the disease spectrum has also undergone significant changes, and coexistence of chronic diseases in arsenicosis has become a primary threat to residents health. In the future, basic study from a new perspective should be deepen, to explore new pathways for integrated prevention and treatment of chronic diseases of arsenicosis, and build a whole management model of "screening, management, prevention, treatment and study" to guarantee the prevention and treatment of chronic diseases of arsenicosis.
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Objective:To investigate the role of DNA damage and repair inhibition in the effect of ginkgo biloba on liver injury in patients with coal-burning-borne arsenism.Methods:In March 2017, the investigation was conducted in Jiaole village arsenic poisoning area in Yuzhang Town, Xingren County, Guizhou Province. According to the "Diagnosis of Endemic Arsenicosis" (WS/T 211-2015) and the "Diagnostic Criteria of Occupational Toxic Hepatopathy" (GBZ 59-2010), 52 patients with arsenism were selected as the ginkgo biloba intervention group, and 49 cases of arsenism patients as intervention control group. Ginkgo biloba tablets were given orally for 3 months (1 tablet/time, 3 times/d) according to the commonly used clinical methods, and no other drugs were given to all subjects during the intervention period. The intervention control group was given placebo in the same way as that of ginkgo biloba intervention group. A total of 41 residents who did not burn high arsenic coal 12 km away with no abnormal liver function were selected as normal control group. Physical examinations were performed before the intervention and at the end of the intervention at 3 months. After receiving signed informed consent, morning urine and peripheral venous blood samples were collected to detect urinary arsenic content by inductively coupled plasma mass spectrometry (ICP-MS); liver function biochemical indexes [albumin (ALB), albumin/globulin (A/G), cholinesterase (CHE), total bile acid (TBA)] were determined by automatic biochemical analyzer, DNA damage by single-cell gel electrophoresis assay, and the expression of miR-145 (repair inhibition index) by qRT-PCR.Results:There were 116 subjects, 41 in normal control group, 39 in ginkgo biloba intervention group and 36 in intervention control group. In ginkgo biloba and intervention and intervention control groups, there was no significant difference in age, gender, smoking habits and drinking compared with normal control group ( P > 0.05). Urinary arsenic content, TBA level, DNA damage degree [comet tail DNA percentage (TailDNA%) and olive tail moment (OTM)] and plasma miR-145 expression level [(38.75 ± 19.09) μg/g Cr, (11.13 ± 1.55) μmol/L, 8.50 ± 0.88, 7.43 ± 0.68, 5.78 ± 0.75, respectively] in ginkgo biloba intervention group patients before intervention were higher than those in normal control group [(11.62 ± 5.33) μg/g Cr, (5.36 ± 0.87) μmol/L, 5.24 ± 0.33, 4.71 ± 0.29, 2.05 ± 0.27, respectively], the differences were statistically significant ( P < 0.05); the levels of ALB, A/G and CHE were significantly lower than those in normal control group ( P < 0.05). After the intervention of ginkgo biloba, urinary arsenic content, TBA level, DNA damage degree (TailDNA% and OTM) and plasma miR-145 expression level in patients were significantly lower than those before the intervention ( P < 0.05); the levels of ALB, A/G and CHE were significantly higher than those before the intervention ( P < 0.05). There was no significant difference in the above indexes before and after intervention in the intervention control group ( P > 0.05). The results of correlation analysis between DNA damage degree, miR-145 and liver function indexes after the intervention of ginkgo biloba showed that, DNA damage degree (TailDNA% and OTM) was negatively correlated with the levels of ALB, A/G and CHE ( r = - 0.34, - 0.33, - 0.48, - 0.31, - 0.31, - 0.42, P < 0.05), and positively correlated with the level of TBA ( r = 0.49, 0.48, P < 0.05); miR-145 was negatively correlated with the levels of ALB, A/G and CHE ( r = - 0.26, - 0.23, - 0.38, P < 0.05), which was positively correlated with the level of TBA ( r = 0.32, P < 0.05); and DNA damage degree was positively correlated with the expression of miR-145 ( r = 0.65, 0.52, P < 0.05). Conclusion:Ginkgo biloba tablets can alleviate the liver damage caused by arsenic through coal burning, and the mechanism of this process is related to its inhibition of miR-145 expression and reduction of DNA damage.
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Objective:To explore the incidence, influencing factors and prognostic value of cardiac valve calcification (CVC) in chronic kidney disease (CKD) non-dialysis patients.Methods:The non-dialysis patients with CKD stage 1-5 who were hospitalized and underwent echocardiography in the Department of Nephrology, Xuanwu Hospital, Capital Medical University from January 1, 2018 to December 31, 2019 were retrospectively admitted. The patients were divided into CVC group and non-CVC group, and the clinical data were compared between the two groups. The deadline for follow-up was November 1, 2021, and the follow-up end point event was all-cause mortality. Logistic regression model was used to analyze the risk factors of CVC in patients with CKD, and Cox proportional hazards regression model was used to analyze the risk factors of all-cause mortality in patients with CKD.Results:A total of 563 patients with CKD were enrolled in the study, with age of (59.49±13.97) years old, and 352 males (62.52%). There were 325 patients (57.73%) with CKD stage 1-3 and 238 patients (42.27%) with CKD stage 4-5. The incidence of CVC in CKD stage 1-5 patients was 32.32%(182/563). Aortic valve calcification occurred in 30.73%(173/563), mitral valve calcification occurred in 9.77% (55/563), double valve (mitral and aortic valve) calcification occurred in 8.35% (47/563), and tricuspid valve calcification occurred in 0.18%(1/563). Age (t=12.223, P<0.001) and the proportions of CKD stage 4-5 ( χ 2=10.854, P=0.001), hypertension ( χ 2=7.811, P=0.005), diabetes ( χ 2=8.424, P=0.004), hyperlipidemia ( χ 2=9.331, P=0.002), and taking statins ( χ 2=4.868, P=0.027) in CVC group were significantly higher than those in non-CVC group. Total cholesterol (t=2.243, P=0.025), low density lipoprotein cholesterol (t=2.025, P=0.043), platelet count (t=2.230, P=0.026) and estimated glomerular filtration rate (t=8.630, P<0.001) in CVC group were lower than those in the non-CVC group. Logistic regression analysis results showed that age≥60 years old (≥60 years old/<60 years old, OR=7.412, 95% CI 4.514-12.170, P<0.001), CKD stage 4-5 (stage 4-5/stage 1-3, OR=2.791, 95% CI 1.730-4.505, P<0.001) and hyperlipidemia ( OR=5.241, 95% CI 3.283-8.367, P<0.001) were the independent influencing factors of CVC in patients with CKD. Five hundred and sixty-three patients were followed up for an average of 26 months, including 68 cases (12.08%) of death, 436 cases (77.44%) of survival and 59 cases (10.48%) of loss to follow-up. Multivariate Cox regression analysis results showed that age≥60 years old (≥60 years old/<60 years old, HR=2.157, 95% CI 1.127-4.127, P=0.020), serum albumin<30 g/L (<30 g/L/≥30 g/L, HR=1.923, 95% CI 1.037-3.568, P=0.038) and double valve calcification (double valve calcification/no valve calcification, HR=2.516, 95% CI 1.279-4.950, P=0.008) were the independent influencing factors of all-cause death in patients with CKD. Conclusions:CVC accounts for 32.32% in non-dialysis patients with CKD stage 1-5. Older age, worse renal function and hyperlipidemia are the independent risk factors of CVC in CKD patients. Older age, hypoproteinemia and double valve calcification are the independent risk factors of all-cause death in patients with CKD.
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Objective:To analyze the incidence and risk factors of acute kidney injury (AKI) in patients undergoing neurosurgery.Methods:This study was a single center and retrospective research. The patients hospitalized in the general neurosurgery ward of Xuanwu Hospital, Capital Medical University, due to intracranial tumors and intracranial vascular diseases from January 1, 2017 to December 31, 2020 were enrolled. Demographic, clinical data and laboratory examination results of the selected patients were collected. The patients were divided into AKI group and non-AKI group according to AKI diagnosis criteria, and the differences of clinical parameters and medication between the two groups were compared. Logistic regression analysis method was used to analyze the risk factors of AKI in neurosurgical patients.Results:Among 4 509 patients undergoing neurosurgery with age of (51.93±16.03) years old, 2 361 males and 2 148 females, 152 patients (3.37%) had AKI. The incidence of AKI in patients undergoing intracranial tumor surgery was 3.69% (84/2 278), and the incidence of AKI in patients undergoing intracranial cerebrovascular surgery was 3.05%(68/2 231). The length of hospital stay ( t=4.897, P<0.001) and operation time ( t=5.496, P<0.001) in AKI group were significantly longer than those in non-AKI group. The proportions of diabetes mellitus, preoperative serum creatinine, blood urea nitrogen, glycosylated hemoglobin, lactic acid, fibrinogen, and systolic pressure levels in AKI group were significantly higher than those in non-AKI group (all P<0.05); the hemoglobin level in AKI group was significantly lower than that in non-AKI group ( P<0.05). The proportions of patients using angiotensin converting enzyme inhibitors/angiotensin Ⅱ receptor antagonists, cephalosporins, proton pump inhibitors, mannitol, and nonsteroidal anti-inflammatory drugs in AKI group were also significantly higher than those in non-AKI group (all P<0.05). Multivariate logistic regression analysis results showed that hemoglobin<110 g/L ( OR=4.252, 95% CI 1.569-11.527, P=0.004), elevated blood urea nitrogen ( OR=1.304, 95% CI 1.139-1.492, P<0.001) and application of nonsteroidal anti-inflammatory drugs ( OR=2.342, 95% CI 1.044-5.253, P=0.039) were independent risk factors of AKI in neurosurgical patients. Conclusions:The incidence of AKI in patients in neurosurgery general ward is 3.37%. Anemia, elevated blood urea nitrogen and application of nonsteroidal anti-inflammatory drugs are independent risk factors of AKI in patients undergoing neurosurgery.
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Background Arsenic is a well-known environmental toxicant. Hepatic fibrosis could occur dueto excessive or long-term exposure to arsenic, while associated molecular mechanisms remain undefined. Mitogen-inducible gene 6 (Mig-6) exhibits a protective effect on numerous diseases or cancers. However, the specific role of Mig-6 in the mechanisms of arsenite-induced hepatic fibrosis remains indistinct. Objective To investigate the specific role of Mig-6 in the activation of hepatic stellate cells (HSC) and the deposition of extracellular matrix (ECM) induced by sodium arsenite (NaAsO2). Methods Human hepatic stellate cells (Lx-2) were treated with 0, 1.875, 3.75, 7.5, and 15 μmol·L−1 of NaAsO2 for 24 h, or with 7.5 μmol·L−1 NaAsO2 for 0, 12, 24, 48, and 72 h. Additionally, Lx-2 cells were transfected by pcDNA3.1(+)/Mig-6, then treated with 7.5 μmol·L−1 NaAsO2 for 24 h; a blank control group, a pcDNA3.1(+)-control group, a pcDNA3.1(+)/Mig-6 group, and an arsenic (7.5 μmol·L−1 NaAsO2) group were also set up. After transfection, the cells and culture supernatants were collected, and the protein levels of Mig-6, α-smooth muscle actin (α-SMA), and transforming growth factor-β1 (TGF-β1) in Lx-2 cells were identified by Western blotting analysis; moreover, the secretion levels of main ECM components in supernatants such as hyaluronic acid (HA), laminin (LN), collagens IV (COL-IV), and procollagen-III (PIIINP) were tested by ELISA. Results The Mig-6 expression decreased in the 3.75, 7.5, and 15 μmol·L−1 NaAsO2 groups (0.561±0.095, 0.695±0.048, and 0.401±0.030) compared to the control group (1.000±0.000) in Lx-2 cells (P<0.05). After administration with 7.5 μmol·L−1 of NaAsO2 for 24, 48, and 72 h, the Mig-6 expression (0.856±0.036, 0.515±0.077, 0.491±0.060) decreased compared with the 0 h group (1.000±0.000) (P<0.05). After over-expression of Mig-6, the results of Lx-2 activation related protein levels showed that compared to the control group, the α-SMA and TGF-β1 expression were up-regulated in the arsenic group (P<0.05); meanwhile, the α-SMA and TGF-β1 in the Mig-6 over-expression combined arsenic exposure group reduced compared to the arsenic (7.5 μmol·L−1) group (P<0.05). The results of ELISA showed that compared with the control group, the HA, LN, PIIINP, COL-IV in the arsenic group were up-regulated (P<0.05); while compared to the arsenic group, the HA, LN, PIIINP, and COL-IV in the Mig-6 over-expression combined with arsenic exposure group were decreased (P<0.05). Conclusion Arsenic down-regulates Mig-6 expression in HSC, and over-expression of Mig-6 can reverse the activation of HSC and ECM deposition induced by arsenic exposure. It suggests that Mig-6 plays a protective role in arsenic-induced HSC activation and ECM deposition.
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Background The key enzymes of serine synthesis pathway (SSP) play an important role in tumor growth, proliferation, and invasion, but their roles in arsenic carcinogenesis are unclear. Objective To observe the effects of NaAsO2 treatment on the expressions of key enzymes [such as phosphoglycerate dehydrogenase (PHGDH), phosphoserine aminotransferase 1 (PSAT1), and phosphoserine phosphatase (PSPH)] of SSP and on the ability to proliferate and migrate in human immortalized skin keratinocytes (HaCaT) and NaAsO2-induced malignantly transformed HaCaT (T-HaCaT), and to explore the roles of SSP key enzymes in arsenic carcinogenesis. Methods (1) The T-HaCaT cells constructed earlier by our research team were divided into a passage control (0 μmol·L−1 NaAsO2) group, a T-HaCaT (0.5 μmol·L−1 NaAsO2) group, a NCT503 (PHGDH inhibitor, 25 μmol·L−1) group, and a NCT503 (25 μmol·L−1) + T-HaCaT (0.5 μmol·L−1 NaAsO2) group. Western blotting was used to detect the protein expression levels of SSP key enzymes in the passage control group and the T-HaCaT group. CCK8 assay and cell scratch test were used to detect the proliferation and migration rates of cells in each group respectively. (2) Well-grown logarithmic-phase HaCaT cells were treated with 0, 0.625, 1.25, and 2.5 μmol·L−1 NaAsO2 for 0, 24, 48, and 72 h to detect cell proliferation rate and protein expression levels of SSP key enzymes. In the subsequent experiment, HaCaT cells were pretreated with 25 μmol·L−1 NCT503 for 6 h, and then treated with 2.5 μmol·L−1 NaAsO2 for 72 h continuously. The experimental groups included a control (0 μmol·L−1 NaAsO2) group, an exposure (2.5 μmol·L−1 NaAsO2) group, a pretreatment (25 μmol·L−1 NCT503) group, and a pretreatment (25 μmol·L−1 NCT503) + exposure (2.5 μmol·L−1 NaAsO2) group, to detect the proliferation rate of cells in each group. Results The protein expression level of PHGDH in the T-HaCaT group were 1.60 times higher than that in the passage control group (P<0.05), and its proliferation rate (177.51%±14.69%) and migration rate (53.85%±0.94%) were also higher than the passage control group’s (100.00%±0.00% and 24.30%±2.26%) (both Ps<0.05), respectively. After the NCT503 intervention, the proliferation rate (144.97%±8.08%) and migration rate (35.80%±0.99%) of cells in the NCT503 + T-HaCaT group were lower than those in the T-HaCaT group (both P<0.05). The proliferation rate of HaCaT cells after NaAsO2 exposure for 72 h increased with the increase of exposure concentration (r=0.862, P<0.05), and consistently, the protein levels of SSP key enzymes in HaCaT cells in each exposure group were higher than those in the control group (all P<0.05). The proliferation rate of HaCaT cells treated with 2.5 μmol·L−1 NaAsO2 increased with the extension of exposure time (r=0.775, P<0.05), which was consistent with the changes of PHGDH levels in cells. After the NCT503 intervention, the proliferation rate of the pretreatment + exposure group was significantly lower than that of the exposure group (P<0.05). Conclusion The key enzymes of SSP may play an important role in the proliferation of T-HaCaT cells induced by NaAsO2.
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Background In addition to the typical signs of skin damage, long-term arsenic exposure is often accompanied by signs and symptoms of neurobehavioral abnormalities. Objective To investigate potential intervention effect of sciadopitysin on senescence of neurons induced by sodium arsenite in rats and possible underlying mediating effect of cell cycle-related transcription factor E2F1. Methods SH-SY5Y cells were treated with 4 μmol·L−1 sodium arsenite for 24 h and intervened with 50 μg·mL−1 Ginkgo biloba extract (EGb761) or four major biflavonoids in Ginkgo biloba leaves (isoginkgetin, bilobetin, sciadopitysin, and ginkgetin) for 24 h respectively. Then, cell viability was measured by CCK-8 assay. Thirty-two 180-200 g SPF rats were randomly divided into a control group, an arsenic treatment group (10 mg·L−1), a Ginkgo biloba extract intervention group (10 mg·kg−1), and a sciadopitysin intervention group (10 mg·kg−1), 8 rats in each group, half male and half female. The rats were treated with sodium arsenite by free drinking water for 3 consecutive months, and the intervention treatment was conducted after 2 months of poisoning with drug intake by gavage for 1 month. HE staining was used to detect structural changes in the hippocampus, while Nissl's staining was used to detect changes in hippocampal morphology and neuron numbers. Moreover, senescence-associated β galactosidase (SA-β-gal) staining and Western blotting were used to detect senescence of hippocampal neurons and the expression level of E2F1, respectively. Results Compared to the arsenic treatment group, EGb761 and the four biflavonoids in Ginkgo biloba leaves effectively antagonized the inhibitory effect of sodium arsenite on cell viability (all Ps<0.05), and sciadopitysin showed better restoration of cellular viability than Ginkgo biloba extract (P<0.05). The results of HE staining and Nissl's staining showed that the hippocampal neurons in the arsenic treatment group were reduced in cell count and the synaptic structure was abnormal, with swelling, nuclear shrinkage, and vacuole, compared with the control group. The results of SA-β-gal staining showed that the number of senescent cells in the arsenic treatment group (15.75±3.01) was significantly increased compared with the control group (2.88±0.84) (P<0.05); the numbers of senescent cells in the Ginkgo biloba extract group (9.38±1.92) and the sciadopitysin treatment group (7.75±2.38) were significantly decreased compared with the arsenic treatment group (all Ps<0.05). The results of Western blotting showed that compared with the control group, the expression of E2F1 protein in hippocampus of the arsenic treatment group was significantly decreased (1.00±0.17 vs. 0.65±0.19, P<0.05); compared with the arsenic treatment group, the protein expression level of E2F1 in hippocampus of the sciadopitysin treatment group (0.89±0.18) was significantly recovered (P<0.05); compared with Ginkgo biloba extract (0.68±0.19), sciadopitysin had a better recovery effect on E2F1 expression level (0.89±0.18) (P<0.05). The results of correlation analysis showed that the E2F1 protein expression level was negatively correlated with the positive rate of SA-β-gal staining in hippocampal neurons (r=−0.518, P<0.05). Conclusion Sciadopitysin is an effective component of Ginkgo biloba extract. It can effectively inhibit the senescence of hippocampal neurons induced by sodium arsenite, and E2F1 may play an important mediating role.
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Background Natural product sanguinarine chloride (SC) can significantly alleviate liver fibrosis and acute liver injury in mice, but whether it has a protective effect on mouse liver injury caused by sodium arsenite (SA) has not been studied. Objective To verify if SC may present preventive and therapeutic effects on SA-induced liver injury in mice. Methods A total of 140 SPF male Kunming mice were randomly divided into two sub-studies, which included a prevention sub-study and a treatment sub-study. In each sub-study, a blank group (normal saline), a model group (5 mg·kg−1 SA), and a positive control group (11.375 mg·kg−1 bicyclol and 182 mg·kg−1 glutathione), as well as SC low, medium, and high dose groups (25, 50, and 100 mg·kg−1) were arranged with 10 mice in each group. In the prevention sub-study, the blank group was given normal saline, the model group was given SA, and the other groups (the SC low, medium, and high dose groups and the positive control group) were given the corresponding treatment 30 min before gavage of SA, once a day, for 28 d. In the treatment sub-study, except for the blank group which was given normal saline, the other groups were given SA for 28 d, then the model group was given normal saline, and the other groups were given the corresponding treatment every day for 28 d. After the experiment, the mice were sacrificed to evaluate selected physiological and biochemical indicators in serum and liver tissue and to observe histopathological changes after HE staining. Results In either sub-study of preventive effect or treatment effect: compared with the blank group, body weight, liver weight, liver coefficient, as well as serum alanine transaminase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), malondialdehyde (MDA), glutathione peroxidase (GSH), and superoxide dismutase (SOD) among all SC groups were not significantly different (P>0.05); but compared with the model group, the SC groups showed increased body weight (P<0.01), decreased liver weight and liver coefficient (P<0.01), reduced ALT, AST, TBIL, and MDA (P<0.05 or P<0.01), and increased GSH and SOD with (P<0.05 or P<0.01) or without significance; compared with the positive control group, no differences were found in the above indicators (P>0.05). The result of histopathological evaluation showed that the SC groups had a clear liver lobule structure, neatly arranged hepatic cords, and less infiltration of inflammatory cells. Conclusion SC has both preventive and therapeutic effects on SA-induced liver injury in mice.
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Background Long-term exposure to arsenic can cause liver injury of varying degrees. Mitochondrial damage may be an early key event of arsenic-induced liver injury. Silent mating type information regulation 2 homolog 1 (SIRT1)/ recombinant peroxisome proliferators-activated receptor gamma coactivator 1 alpha (PGC-1α) is an important pathway regulating mitochondrial mass and function. However, whether arsenic-induced liver injury is related to mitochondrial dysfunction mediated by SIRT1/PGC-1α pathway remains unclear. Objective To investigate potential effects of sodium arsenite (NaAsO2) on mitochondrial function and expressions of SIRT1/PGC-1α pathway-related proteins in human normal liver cell. Methods Human normal liver cells (MIHA cells) were used as the research object. MIHA cells were treated with different concentrations of NaAsO2 (0, 5, 10 and 20 μmol·L−1) for 24 h, and the cells were collected for study. The ultrastructure of mitochondria was observed by transmission electron microscopy, adenosine triphosphate (ATP) concentration by fluorescence method, mitochondrial membrane potential (MMP) level by flow cytometry, and SIRT1, PGC-1α and their downstream nuclear respiratory factor 1 (NRF1) and mitochondrial transcription factor A (TFAM) protein expression levels by Western blotting. One-way analysis of variance and trend test were used for data statistical analysis. Results The viability of MIHA cells decreased gradually with the increase of NaAsO2 concentration (F=6495.47, P<0.001). The transmission electron microscope observation showed that the size of mitochondria in the 10 μmol·L−1 NaAsO2 treatment group was different, and the mitochondria were swollen or elongated in a rod-like shape. The mitochondria in the 20 μmol·L−1 NaAsO2 treatment group swelled like air spheres or vacuoles. The ATP concentration and MMP level of MIHA cells gradually decreased with the increase of NaAsO2 concentration (Ftrend of ATP=172.28, Ftrend of MMP=59.91, both Ps<0.001). Compared with the control group, the protein expression levels of SIRT1, PGC-1α, NRF1, and TFAM were not significantly changed in the 5 μmol·L−1 NaAsO2 treatment group, while the protein expression levels of SIRT1, PGC-1α, and TFAM were decreased in the 10 μmol·L−1 NaAsO2 treatment group, and the protein expression levels of SIRT1, PGC-1α, and NRF1 were decreased in the 20 μmol·L−1 NaAsO2 treatment group. The results of trend test showed that the protein expression levels of SIRT1, PGC-1α, NRF1, and TFAM decreased gradually with the increase of NaAsO2 concentration (Ftrend of SIRT1=47.07, P<0.001; Ftrend of PGC-1α=15.17, P<0.01; Ftrend of NRF1=13.54, P<0.01; F trend of TFAM=4.20, P<0.05). Conclusion The down-regulation of SIRT1/PGC-1α and its downstream NRF1 and TFAM may be involved in NaAsO2-induced mitochondrial dysfunction in liver cells.
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Objective:To investigate the role of liver X-activated receptor (LXRα)/sterol-regulatory element binding protein (SREBP-1c) in arsenic induced lipid metabolism disorders in rats, and to provide a basis for study the mechanism of arsenic induced lipid metabolism disorders.Methods:Twenty-four healthy clean grade Wistar rats, were randomly divided into 4 groups according to body weight (80 - 100 g) by the random number table method, with 6 rats in each group, half male and half female. Rats in control group were given deionized water by gavage. The low, medium and high arsenic dose groups were given 2.5, 5.0 and 10.0 mg·kg -1·d -1 sodium arsenite solution by gavage, respectively. They were exposed to arsenic for 6 days a week for 4 months. At the end of the experiment, blood and liver samples of rats in each group were collected. The hepatic arsenic content was determined by inductively coupled plasma mass spectrometry (ICP-MS); the serum levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were measured by automatic biochemical analyzer. The mRNA expression levels of LXRα and SREBP-1c in liver tissues were determined by real-time PCR; the protein expression levels of LXRα, SREBP-1c, acetyl CoA carboxylase (ACC) and phospho-ACC (pACC) in liver tissues were determined by Western blotting. Results:The hepatic arsenic contents of rats in low, medium and high arsenic dose groups were (61.04 ± 4.98), (62.66 ± 6.71) and (87.86 ± 13.89) μg/g, respectively, which were higher than that in control group [(2.43 ± 0.63) μg/g, P < 0.05], and the hepatic arsenic content of rats in high arsenic dose group was higher than those in low and medium arsenic dose groups ( P < 0.05). The serum TG levels of rats in low, medium and high arsenic dose groups were (0.90 ± 0.17), (1.28 ± 0.24) and (1.82 ± 0.18) mmol/L, respectively, which were higher than that in control group [(0.50 ± 0.12) mmol/L, P < 0.05]; the serum LDL-C levels of rats in low, medium and high arsenic dose groups were (0.54 ± 0.04), (0.63 ± 0.07) and (0.69 ± 0.08) mmol/L, respectively, which were higher than that in control group [(0.27 ± 0.05) mmol/L, P < 0.05]; the serum TC levels of rats in medium and high arsenic dose groups were (1.88 ± 0.23) and (2.10 ± 0.10) mmol/L, respectively, which were higher than that in control group [(1.51 ± 0.14) mmol/L, P < 0.05]; the serum HDL-C levels of rats in medium and high arsenic dose groups were (0.84 ± 0.11) and (0.71 ± 0.14) mmol/L, respectively, which were lower than that in control group [(1.15 ± 0.08) mmol/L, P < 0.05]; and the serum levels of TG and LDL-C in medium and high arsenic dose groups were higher than those in low arsenic dose group ( P < 0.05), and the serum level of TG in high arsenic dose group was higher than that in medium arsenic dose group ( P < 0.05). The mRNA expression level of hepatic LXRα of rats in high arsenic dose group was higher than those in control group and low arsenic dose group ( P < 0.05); there was no significant difference in mRNA expression levels of hepatic SREBP-1c of rats between low, medium and high arsenic dose groups and control group ( P > 0.05). The protein expression levels of hepatic LXRα of rats in medium and high arsenic dose groups were higher than that in control group ( P < 0.05), and high arsenic dose group was higher than low arsenic dose group ( P < 0.05); the protein expression levels of hepatic SREBP-1c and ACC of rats in high arsenic dose group were higher than that in control group ( P < 0.05). There was a positive correlation between hepatic arsenic content in arsenic-exposed rats and the serum levels of TG, TC, LDL-C, the mRNA expression level of hepatic LXRα, the protein expression levels of hepatic LXRα, SREBP-1c and ACC ( r = 0.84, 0.62, 0.89, 0.55, 0.54, 0.64, 0.70, P < 0.05), and the serum level of HDL-C was negatively correlated with the hepatic arsenic content in arsenic-exposed rats ( r = - 0.75, P < 0.001). Conclusion:Sodium arsenite can increase the serum levels of TG, TC and LDL-C, decrease the serum level of HDL-C and increase the protein expression levels of LXRα and SREBP-1c in liver tissues, suggesting that arsenic induced lipid metabolism disorders in rats may be related to the upstream regulation mechanism of LXRα/SREBP-1c.