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Chinese Journal of Blood Purification

    12 December 2023, Volume 22 Issue 12 Previous Issue    Next Issue
    Perioperative Management of New-built Autogenous Arteriovenous Fistula: Chinese Expert Consensus
    Expert Consensus Work Group of Nephrology and Blood Purification
    2023, 22 (12):  881-890.  doi: 10.3969/j.issn.1671-4091.2023.12.001
    Abstract ( 316 )   PDF (583KB) ( 48 )  
    Autogenous arteriovenous fistula (AVF) is the first choice for vascular access in hemodialysis patients. Detailed evaluation and planning for a new AVF before the surgery, meticulous manipulation during surgery, and close monitoring and exercise guidance after the surgery must be carried out. These processes need mutual interaction and collaboration between doctors and patients. Healthcare education should begin before the use of AVF for hemodialysis. The Expert Consensus Work Group of Nephrology and Blood Purification, Beijing Perioperative Medicine Study Society based on the guidelines, consensus and evidence-based medical data from China and foreign countries and combined with the clinical experiences of the members in this group proposes this consensus to provide suggestions and comments about the perioperative management of AVF.
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    Analysis of the factors for vascular endothelial dysfunction after low sodium dialysis in maintenance hemodialysis
    SHEN Jian, XIE Yi-nv, YE Ling, HUANG Li, WANG Wei-ying, PANG Jun
    2023, 22 (12):  891-895.  doi: 10.3969/j.issn.1671-4091.2023.12.002
    Abstract ( 70 )   PDF (645KB) ( 16 )  
    Objective  To investigate the factors affecting vascular endothelial dysfunction (VED) after hemodialysis with low sodium dialysate in chronic kidney disease (CKD) patients.  Methods The CKD patients undergoing maintenance hemodialysis (MHD) in Yuyao People's Hospital from January 2019 to February 2023 were included in this study. The sodium concentration in dialysate was set at 137mmol/L. The percentage of increased reactive diastolic diameter in brachial artery was compared at baseline and after 6 months of dialysis with low sodium dialysate. Logistic regression was used to analyze the risk factors for VED. A nomograph model was built and validated with self-sampling method. The discrimination and accuracy of the model were evaluated by receiver operating characteristic (ROC) curves and calibration curves. Results A total of 80 CKD patients were enrolled, including 42 with mild VED and 18 with severe VED. Multifactorial logistic regression showed that dialysis age (OR=1.429, 95% CI: 1.241~1.657, P<0.001), brachial artery blood flow (OR=1.611, 95% CI: 1.472~1.713, P<0.001), hypersensitive C-reactive protein (OR=1.376, 95% CI: 1.194~1.582, P<0.001), and malonic dialdehyde (OR=1.376, 95% CI: 1.194~1.582, P<0.001) were the independent risk factors for VED; albumin (OR=0.472, 95% CI: 0.318~0.609, P<0.001) and superoxide dismutase (OR=0.658, 95% CI: 0.523~0.791, P<0.001) were the protective factors for VED. The nomograph showed a high discrimination ability in predicting the risk of VED. The area under the ROC curve was 0.889 (95% CI: 0.841~0.937); the sensitivity was 90.79% and the specificity was 80.73%. The calibration curve showed a good fit between predicted and actual probabilities.  Conclusion  Longer dialysis age, higher brachial artery blood flow, higher serum hypersensitive C-reactive protein, higher serum malonic dialdehyde, low serum albumin, and lower serum superoxide dismutase are the risk factors for VED after hemodialysis with low sodium dialysate in CKD patients. The nomograph model can be used for close monitoring and early warning of these risk factors.
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    Analysis of the influencing factors for skin itching in uremic hemodialysis patients and construction of a column chart model for prediction analysis
    U Yao-ling, ZHONG Li-li, ZHOU Fang, GU Hong-ye
    2023, 22 (12):  896-899.  doi: 10.3969/j.issn.1671-4091.2023.12.003
    Abstract ( 113 )   PDF (650KB) ( 45 )  
    Objective  To explore the influencing factors for skin itching in uremic hemodialysis patients and to construct a column chart model for prediction analysis.  Methods  A total of 162 uremic hemodialysis patients admitted to Huai'an Second People's Hospital (Huai'an Hospital affiliated to Xuzhou Medical University) from July 2020 to December 2022 were recruited as the study subjects. They were grouped into a skin itching group (96 cases) and a non-skin itching group (66 cases) based on the symptom of skin itching. Clinical data were collected from the patients. Logistic regression was applied to analyze the risk factors for skin itching in uremic hemodialysis patients. R software was applied to construct a column chart model, and calibration curve was applied to verify the consistency of the mode. The area under the ROC curve was applied to evaluate the predictive value and discrimination ability of the model.  Results  Ninety-six cases (59.26%) of the 162 uremic hemodialysis patients had pruritus. Univariate analysis showed that diabetes history (χ2=5.569, P=0.018), parathyroid hormone (t=13.510, P<0.001), β2 microglobulin (t=11.836, P<0.001), serum phosphorus (t=10.478, P=0.001) and C-reactive protein (t=13.221, P<0.001) were significantly different between skin itching group and non-skin itching group. Multivariate logistic regression showed that β2 microglobulin (OR=1.451, 95% CI: 1.217~1.731, P<0.001), serum phosphorus (OR=7.455, 95% CI: 1.194~46.533, P=0.032), and C-reactive protein (OR=10.929, 95% CI: 4.086~29.234, P<0.001) were the independent risk factors for itching in uremic hemodialysis patients. The nomogram prediction model had better differentiation (area under ROC curve=0.977, 95% CI:0.956~0.998) and consistency (Hosmer-Lemeshow goodness of fit test, χ2=10.065, P=0.261) abilities.  Conclusion   This study constructs a column chart model for predicting the risk of skin itching in uremic hemodialysis patients based on three risk factors: β2 microglobulin, serum phosphorus, and C-reactive protein. This model has better discrimination and consistency abilities, and is expected to become an important tool for predicting the risk of skin itching in uremic hemodialysis patients.
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    The clinical characteristics and outcome of peritoneal dialysis associated peritonitis by multiple drug resistant organisms
    YANG Bin-hong, WANG Xiu-fen, ZHANG Tao, LI Ying
    2023, 22 (12):  900-904.  doi: 10.3969/j.issn.1671-4091.2023.12.004
    Abstract ( 52 )   PDF (559KB) ( 11 )  
    Objective  To investigate the clinical characteristics, bacterial spectrum, drug resistance rate and outcome of peritoneal dialysis-associated peritonitis (PDAP) by multiple drug resistant organisms (MDRO), so as to provide clinical evidences for standardizing therapy of PDAP by MDRO.  Methods  Clinical data of PDAP patients admitted to The Third Hospital of Hebei Medical University during January, 2016 and December, 2022 were retrospectively reviewed. They were divided into two groups based on the results of bacterial culture in effluent dialysate, PDAP by MDRO group and PDAP not by MDRO group. Clinical data and treatment results were compared between the two groups.  Results  ①In the 96 PDAP patients, a total of 123 PDAP episodes occurred, of which 56 episodes were PDAP by MDRO. The MDRO were dominated by Gram positive bacteria (49/56, 87.5%), in which the most strains were coagulase-negative staphylococci (41/49, 83.7%); all Gram negative bacteria were Escherichia coli (E. coli). ②The average time for significant improvement of PDAP was longer in PDAP by MDRO group than in PDAP not by MDRO group (Z=-1.994 and -2.341; P=0.046 and 0.019); the average time for significant improvement of PDAP caused by Gram positive bacteria was 2.491 days longer in PDAP by MDRO group than in PDAP not by MDRO group (t=-2.254, P=0.028). ③The sensitivity of Gram positive bacteria to rifampicin increased in 2020~2022 compared to that in 2016-2019 (χ2=4.416, P=0.042).  ④In the PDAP caused by Gram positive bacteria, the recurrence rate of PDAP was higher in PDAP by MDRO group than in PDAP not by MDRO group (P=0.020).  Conclusion  The main pathogens in PDAP by MDRO group were coagulase-negative staphylococci. In PDAP patients by MDRO, the recurrence rate of PDAP was higher, the presence of PDAP symptoms was longer, and effective antibiotic therapy should be used earlier and longer.
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    Relationship between phase lag index and cognitive dysfunction in elderly hemodialysis patients
    WANG Min, LI Zuo-lin
    2023, 22 (12):  905-908, 924.  doi: 10.3969/j.issn.1671-4091.2023.12.005
    Abstract ( 41 )   PDF (564KB) ( 11 )  
    Objective  To investigate the relationship between phase lag index (PLI) and cognitive impairment (CI) in elderly maintenance hemodialysis (MHD) patients.  Methods  A total of 100 elderly patients undergoing regular hemodialysis in our hospital from August 2019 to June 2023 were selected and divided into CI group (n=61) and non-CI group (n=39) according to the Montreal Cognitive Assessment (MoCA) score. All patients underwent electroencephalogram (EEG) examination and PLI was calculated. The clinical data and PLI of various frequency bands were compared between the two groups. Pearson correlation was used to analyze the relationship between PLI and MoCA score. Multivariate logistic regression was used to analyze the risk factors for CI in elderly MHD patients. The receiver operating characteristic (ROC) curve was used to analyze the predictive value of PLI for CI in elderly MHD patients.  Results  The α band PLI was significantly lower in CI group than in non-CI group (t=7.585, P<0.001). There were statistically significant differences in α band PLI from C3-P3, C4-P8, F3-C5, F4-P3, F8-P8, P4-CP3, CP3-O1 and CP5-O2 electrode pairs between the two groups (t=10.692, 6.485, 5.507, 7.505, 7.342, 14.720, 8.413 and 6.947 respectively; P<0.001). Correlation analyses showed that there was a positive correlation between α-band PLI and MoCA score in CI group (R=0.486, P=0.013). Multivariate logistic regression showed that MoCA score (OR=11.512, 95% CI: 2.159~61.389, P=0.008) and α-band PLI (OR=5.669, 95% CI: 1.595~20.149, P=0.023) were the risk factors for CI in elderly MHD patients. ROC curve analyses showed that the area under the curve (AUC) of α-band PLI in predicting CI in elderly MHD patients was 0.818 (95% CI: 0.747~0.888), with the sensitivity and specificity of 78.46% and 64.62% respectively.  Conclusion  The decrease of α-band PLI is closely related to the decline of cognitive function in elderly MHD patients. α-band PLI has a better efficacy in predicting CI in elderly MHD patients and has the significance of guidance for clinical evaluation of CI.
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    Construction and external validation of a risk prediction model for hospitalization and mortality in hemodialysis patients with heart failure
    TANG Wen-wu, YUAN Xin-zhu, YANG Xiao-hua, CHEN Xiao-xia, WANG Zhi-xin, ZHANG Ying, XIE Xi-sheng
    2023, 22 (12):  909-915.  doi: 10.3969/j.issn.1671-4091.2023.12.006
    Abstract ( 51 )   PDF (1126KB) ( 10 )  
    Objective  To construct a nomogram to predict the risk of hospitalization for heart failure (HF) in maintenance hemodialysis (MHD) patients.  Methods  MHD patients from four centers in northeast Sichuan during 2017 to 2023 were included in this study. Their basic information, clinical data, laboratory and imaging results were collected. Patients in the two centers were used as the training set (n=386), and those in the other two centers were used as the external validation set (n=116). Least absolute shrinkage and selection operator (LASSO) and Cox regression analysis were used to analyze the related risk factors. A nomogram model for the risk of HF hospitalization was established. The prediction efficiency of the model was evaluated by the area under the receiver operating characteristic (ROC) curve, and the accuracy and practicability of the model were analyzed and evaluated by the calibration curve and the decision curve.  Results  The median follow-up periods of the training set and external validation set were 15 months (9, 24) and 14 months (10, 21), respectively. HF hospitalization occurred in 140 cases (36.27%) and 28 cases (24.14%) in training set and external validation set, respectively. Cox regression showed that the N-terminal pro-brain natriuretic peptide (HR=1.532, 95% CI:1.244~1.886, P<0.001), percentage of lymphocytes (HR=0.975, 95% CI:0.952~0.999, P=0.038), right atrium diameter (HR=1.060, 95% CI:1.017~1.105, P=0.005)/right ventricle diameter (HR=1.033, 95% CI:0.998~1.062, P=0.064), weekly dialysis duration (HR=0.667, 95% CI:0.459~0.968, P=0.033), HF score (HR=1.778, 95% CI:1.130~2.798, P=0.013), use of angiotensin-converting enzyme inhibitor/angiotensin receptor blocker medications (HR=0.569, 95% CI: 0.353~0.917, P=0.020), and history of coronary heart disease/diabetes (HR=1.582, 95% CI:1.002~2.500, P=0.049) were independently associated with HF hospitalization. The C-statistic for internal and external validation were 0.836 (95% CI:0.802~0.870) and 0.819 (95% CI:0.786~0.853), respectively. The calibration curve showed that there was a good consistency between actual probability and predicted probability of HF hospitalization (the median of the 2-year calibration slope was 1.018). The decision curve showed that the clinical net income was higher.  Conclusion  The prediction model of this study can accurately and individually evaluate the risk of HF hospitalization in MHD patients, and is of clinical practice value.
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    Research progresses in inflammatory factors in hemodialysis patients complicated with depression
    ZHANG Shun-juan, XIA Yun-feng
    2023, 22 (12):  916-919.  doi: 10.3969/j.issn.1671-4091.2023.12.007
    Abstract ( 66 )   PDF (437KB) ( 13 )  
    Hemodialysis is the most important replacement treatment for chronic kidney disease progressed to end-stage renal disease. The incidence of depression has been reported to be 20~55% in hemodialysis patients. Recent studies have shown that inflammatory factors play important roles in hemodialysis patients, and have impacts on the presence and development of depression. This article reviews the research progresses in inflammatory factors in the pathogenesis of depression in hemodialysis patients and its treatment.
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    Strategies for blood purification in sepsis
    ZHANG Qing-qing, HAN Xu-dong
    2023, 22 (12):  920-924.  doi: 10.3969/j.issn.1671-4091.2023.12.008
    Abstract ( 105 )   PDF (517KB) ( 12 )  
    Sepsis is complicated with dysregulation of immune system to infection, which may lead to life-threatening dysfunction of organs. In the presence of infections, pathogen-associated molecular patterns (PAMPs) initiate inflammatory responses to clear the pathogens, but hyper-activated responses of the immune system may injure the host cells and organs. Damage-associated molecules released by the injured cells amplify the inflammatory responses and may progress to develop a cytokine storm through the damage-associated molecular patterns (DAMPs) pathway. In sepsis, neutrophils shift from potent antimicrobial protectors to destructive mediators to tissues and organs. Blood purification removes the inflammatory mediators as well as the immune cells with adverse functions, and becomes an important approach for the treatment of sepsis. Here we summarize the roles of lipopolysaccharides, cytokines, DAMPs and neutrophils in the pathogenesis of sepsis. We also introduce a series of blood purification methods and their clinical trials targeting the clearance of endotoxins, cytokines, and immune cells at the upstream of inflammatory cascade in sepsis.
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    Research progresses of different blood purification methods in the perioperative period of kidney transplantation
    WANG Xin-ran, WANG Jian-wen
    2023, 22 (12):  925-928.  doi: 10.3969/j.issn.1671-4091.2023.12.009
    Abstract ( 54 )   PDF (426KB) ( 11 )  
    Renal transplantation is the best renal replacement therapy for uremic patients. A successful kidney transplantation often relies on blood purification, and different blood purification methods also affect the prognosis of kidney transplant patients. Therefore, it is particularly important to explore the impact of various blood purification techniques in the perioperative period of kidney transplantation. This article reviews the research progresses of various blood purification methods in the perioperative period of kidney transplantation.
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    Application of high cut-off dialysis for severe acute kidney injury secondary to cast nephropathy in patients with multiple myeloma
    LI Jing-jing, ZHANG Wen-jun, TANG Ya, ZHAO Peng-ming
    2023, 22 (12):  929-933.  doi: 10.3969/j.issn.1671-4091.2023.12.010
    Abstract ( 55 )   PDF (533KB) ( 11 )  
    Multiple myeloma (MM) is the second commonest hematological malignant tumor. One of the most important and commonest organs damaged in MM is kidney, usually manifested as myeloma cast nephropathy (MCN) caused by a large number of serum free light chains (sFLCs) from malignant plasma cells. MCN is difficult to treat with poor prognosis, and is one of the main causes of early death in MM. Rapid reduction of sFLCs may improve the kidney function. High cut-off hemodialysis (HCO-HD) with the filters of large pore size can effectively clear the excessive sFLCs. HCO-HD combined with chemotherapy such as bortezomib lead to a tendency of better renal prognosis, but the economic benefit is yet controversial. The results of HCO-HD for MCN is inconsistent, if comparison is made with the results using high flux hemodialysis (HFHD). Currently, there is no standard HCO-HD protocol, and large-scale samples, long-term follow-up, and randomized controlled trials are required to evaluate the clinical outcomes of MCN patients treated by HCO-HD. This article reviews the pathogenesis of MCN, and the clinical application of HCO-HD in MCN.
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    Effect of perivascular adipose tissue on the patency of arteriovenous fistula in hemodialysis patients
    LI Jie, HOU Guo-cun, FENG Guo-hui, JIANG Lin-sen
    2023, 22 (12):  934-938.  doi: 10.3969/j.issn.1671-4091.2023.12.011
    Abstract ( 50 )   PDF (489KB) ( 12 )  
    Objective  To evaluate the regulatory effects of perivascular adipose tissue (PVAT) volume around autologous arteriovenous fistulas (AVF) and fat attenuation index (FAI) on vascular remodeling in patients with AVF.  Methods  A total of 90 patients undergoing AVF surgery in the three clinical trial centers were enrolled in this study. Fat volume and FAI around the AVF in the area of vessel length equal to the vessel diameter (d) were measured by CT at the second postoperative day. AVF patency after the surgery for one year was assessed by Doppler ultrasound. The patients were then divided into non-adverse event group and adverse event group according to the Chinese Expert Consensus on Vascular Access for Hemodialysis, 2nd edition. Multivariate logistic regression model was used to analyze the impacts of PVAT volume and FAI on AVF patency in one year after surgery.  Results  Patients in adverse event group (n=21) had less PVAT volume (t=6.572, P=0.018) and higher FAI (t=-4.883, P=0.025) as compared with those in non-adverse event group (n=61). Logistic regression showed that fat volume (OR=0.256, 95% CI:0.087~0.752, P=0.013) and FAI (OR=1.064, 95% CI:1.007~1.124, P=0.026) were the independent predictive factors for AVF adverse events.  Conclusion  PVAT volume was the protective factors and FAI was the risk factor for AVF patency. Patients with high risk of AVF adverse events can be identified by early postoperative screening, so as to decrease the negative factors and improve the prognosis of AVF.
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    Survival analysis of arteriovenous graft in maintenance hemodialysis patients: a retrospective cohort study
    ZHU Wei, JIANG Jun, PENG Li, WANG Peng, ZHU Cheng-xiu, LAN Lei
    2023, 22 (12):  939-943.  doi: 10.3969/j.issn.1671-4091.2023.12.012
    Abstract ( 58 )   PDF (581KB) ( 10 )  
    Objective  To assess the primary patency rate and secondary patency rate of arteriovenous graft (AVG), and to analyze the risk factors for survival of AVG in maintenance hemodialysis (MHD) patients.  Methods  Baseline data of the MHD patients undergoing AVG surgery at the Department of Nephrology, Anhui Provincial Hospital from December 2016 to June 2022 were retrospectively recruited. Patients were followed up for survival of AVG and prognosis of the patients. Kaplan-Meier method was used to calculate technical survival rate of AVG. Cox regression model was used to analyze the risk factors for primary patency and secondary patency of AVG. The risk ratio (HR) and 95% confidence interval (CI) were calculated for every risk factor.  Results  A total of 51 MHD patients undergoing AVG surgery were collected, and 43 of the 51 patients were finally enrolled in this study because of loss of follow-up in 8 patients. The primary patency and secondary patency periods were 6 (1, 12) and 8 (2, 30) months respectively in the 43 patients. After the surgery for 6, 12 and 18 months, the primary patency rates were 53.9%, 29.0% and 19.9% respectively, and the secondary patency rates were 80.8%, 71.1% and 67.1% respectively. Univariate Cox regression showed that history of central venous catheterization (HR 2.091, 95% CI: 1.013~4.319, P=0.046) was the risk factor for primary patency rate of AVG, and female was the risk factor for secondary patency rate of AVG (HR 3.085, 95% CI:1.022~9.305, P=0.046).  Conclusion  The primary patency and secondary patency periods of AVG were shorter and the long-term patency rate of AVG was lower, indicating the difficulties for a higher long-term patency rate of AVG. Central vein catheterization should be avoided in patients before AVG surgery. AVG should be carefully used for blood access in female MHD patients, Close follow-up after the surgery is required to treat complications in time to improve the secondary patency rate.
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    Relationship between the degree of vascular calcification and arteriovenous fistula dysfunction in hemodialysis patients
    LIN Liu-ping, BI Hui-xin, QIN Xin-fang
    2023, 22 (12):  944-948.  doi: 10.3969/j.issn.1671-4091.2023.12.013
    Abstract ( 55 )   PDF (704KB) ( 11 )  
    Objective  To investigate the relationship between the degree of vascular calcification (VC) and arteriovenous fistula (AVF) dysfunction in patients with maintenance hemodialysis (MHD).  Methods   A total of 166 patients subjected to standard arteriovenous fistula surgery for the first time in the Department of Nephrology, The Affiliated Hospital of Guilin Medical College from January 2019 to January 2020 were selected. Their general clinical information and laboratory examinations were collected. According to the coronary artery calcification score (CACS) at the time of AVF surgery, the patients were divided into non-calcification group, mild calcification group, moderate calcification group and severe calcification group. The clinical factors relating to CACS and the risk factors for AVF dysfunction were analyzed. The primary patency rate of AVF in 2 years in the patients with and without VC was observed.  Results  A total of 166 MHD patients were enrolled including 92 cases in no calcification group, 33 cases in mild calcification group, 19 cases in moderate calcification group, and 22 cases in severe calcification group. Univariate analysis showed that age (H=4.877, P=0.017), diabetes (c2=9.083, P=0.028), diastolic blood pressure (H=17.037, P=0.001), serum calcium (H=12.260, P=0.007) and intact parathyroid hormone (H=9.698, P=0.021) were statistically different among the four groups (P<0.05). Multivariate Cox regression showed that venous diameter (HR=0.496, 95% CI:0.317~0.777, P=0.001) was an protective factor for AVF dysfunction. Spearman correlation analysis showed that AVF dysfunction was positively correlated with BMI(r=0.214,P=0.006), albumin(r=0.173,P=0.026), platelet/lymphocyte ratio (PLR r =0.214,P=0.006) and venous diameter(r=0.231,P=0.003). The patency rates were 71.69%, 60.24%, 50% and 40.96% after the surgery for 6 months, 12 months, 18 months and 24 months. Kaplan-Meier survival curve of the primary patency period showed that VC did not affect the patency rate of AVF (log rank test, χ2=2.407, P=0.121).  Conclusions   The degrees of coronary artery calcification had no effect on AVF dysfunction in two years after surgery.
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    Effect of ultrasound-guided cannulation of arteriovenous fistula in hemodialysis patients: a Meta-analysis
    Effect of ultrasound-guided cannulation of arteriovenous fistula in hemodialysis patients: a meta-analysis
    2023, 22 (12):  949-954.  doi: 10.3969/j.issn.1671-4091.2023.12.014
    Abstract ( 56 )   PDF (836KB) ( 21 )  
    Objective  To systematically evaluate the effect of ultrasound-guided cannulation of arteriovenous fistula (AVF) in hemodialysis patients.  Methods  The databases of PubMed, Embase, Cochrane Library, CNKI, VIP and Wanfang were searched for randomized controlled trials (RCTs) about the efficacy comparison between ultrasound-guided puncture and traditional blind puncture in AVF from the inception to October 2022. Meta-analysis was performed using Revman 5.0 and Stata 16.0 software.  Results  A total of 8 RCTs with 281 patients and 4992 punctures were included. Meta-analysis showed that ultrasound guidance could improve the success rate of one puncture (RR=1.12, 95% CI:1.05~1.20, P<0.001), reduce the incidence of complications after puncture (RR=0.30, 95% CI:0.13~0.71, P=0.006), and extended the pre-puncture evaluation time (MD=75.26, 95% CI:68.16~82.36, P<0.001); but the time for puncture (MD=12.91, 95% CI:-11.46~37.28, P=0.300) and pain perception (MD=-0.11, 95% CI:-0.67~0.45, P=0.710) had no statistical significance.  Conclusion  Compared with traditional blind puncture, ultrasound-guided AVF puncture can significantly improve the success rate of one puncture and reduce the incidence of complications. However, its impacts on puncture time, evaluation time and pain perception needs to be verified by further randomized controlled studies with larger samples.
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