Objective:Pyroptosis is a new type of programmed cell death associated with many inflammatory diseases. Polysaccharides have anti-inflammatory effects. In this study, we investigated whether corn silk polysaccharides (DCSP) before and after selenization (Se-DCSP) can reduce the renal tubule pyroptosis induced by calcium oxalate crystals. Methods:HK-2 cells were exposed to calcium oxalate monohydrate with a size of 3 µm (COM-3μm) to establish a pyroptosis model. The degree of cell damage was determined by detecting cell viability, reactive oxygen species (ROS), and lactate dehydrogenase (LDH) content. The proportion of pyroptosis cells was quantitatively detected by Caspase-1/PI double staining. The expression levels of NLRP3, GSDMD, IL-18, and IL-1β were detected by confocal microscopy and Western blot analyses. Results:DCSP and Se-DCSP can reduce the secretion of inflammatory factors IL-1β/18 related to pyroptosis by reducing cell damage and oxidative stress, as well as down-regulate the expression of Caspase-1, NLRP3, GSDMD, and TNF-α, repair damaged cells, and inhibit pyroptosis in HK-2 cells. The inhibitory effect of selenized polysaccharide was significantly enhanced compared with that before selenification. Conclusion:Se-DCSP can inhibit pyroptosis through the NLRP3/Caspase-1/GSDMD/IL-1β/IL-18 signaling pathway to reduce the risk of kidney-stone formation.

End-stage renal disease（ESRD） patients undergoing outpatient hemodialysis（HD） and home peritoneal dialysis（PD） are high risk population of severe and critical types caused by SARS-CoV-2 infection. In order to improve the quality of diagnosis and treatment in dialysis patients with SARS-CoV-2 infection, we wrote this recommendation for primary care clinicians. During the epidemic period of SARS-CoV-2 infection, all patients should be instructed to strengthen self-management. Once the SARS-CoV-2 infection was found in dialysis patients, early stratified management should be carried out within 72 hours after the first positive nucleic acid or antigen test results, which includes early antiviral therapy, early recognition, and transferring severe patients from community or primary hospital to a referral hospital promptly. Guidance for dietary and sports rehabilitation after SARS-CoV-2 infection should also be started as soon as possible.

目的:研究补体C3a受体(C3aR)在尿路感染模型小鼠膀胱和肾脏的表达与分布.方法:将雌性C57BL/6小鼠随机分为正常对照组,以及3、6、24和48 h感染组,每组8～10只.采用逆行推注人型大肠杆菌J96建立小鼠尿路感染模型.平板涂布计数法检测小鼠膀胱和肾脏细菌负荷量;HE染色评估膀胱和肾脏病理损伤程度;ELISA检测肾组织肾脏损伤分子1(KIM-1)和尿液中C3a水平;蛋白印迹和免疫荧光检测C3aR在膀胱和肾脏的表达与分布.结果:感染24 h时小鼠膀胱细菌负荷量达到峰值,48 h时细菌量下降;感染6 h时小鼠肾脏中细菌负荷量最高,随着感染时间延长逐渐减少.感染后,膀胱病理表现为移行上皮细胞脱落,黏膜和固有层炎症细胞浸润;肾脏病理表现为肾小球萎缩,肾小管上皮细胞损伤,肾小管间质炎症细胞浸润及肾盂处组织脱落.感染后肾脏KIM-1显著增加,尿液中C3a水平升高.蛋白印迹结果显示,感染后膀胱和肾脏C3aR蛋白水平升高.免疫荧光分析显示,C3aR在正常膀胱上皮细胞有少量表达,感染后在上皮细胞大量表达,在深层肌肉组织有少量表达;C3aR在正常肾皮质的肾小球和髓质的实质细胞有少量表达,感染后在肾小球、肾小管间质以及髓质的炎症浸润细胞中大量表达.结论:小鼠尿路感染过程中C3a-C3aR信号激活,C3a在尿液中表达升高,C3aR在膀胱和肾脏中表达上调,炎症细胞表达的C3aR参与小鼠尿路感染进程并发挥作用.

Nano-hydroxyapatite (nano-HAP) is often used as a crystal nest to induce calcium oxalate (CaOx) kidney stone formation, but the mechanism of interaction between HAP crystals of different properties and renal tubular epithelial cells remains unclear. In this study, the adhesion and endocytosis of HAP crystals with sizes of 40 nm, 70 nm, 1 μm, and 2 μm (HAP-40 nm, HAP-70 nm, HAP-1 μm, and HAP-2 μm, respectively) to human renal proximal tubular epithelial cells (HK-2) were comparatively studied. The results showed that HAP crystals of all sizes promoted the expression of osteopontin and hyaluronic acid on the cell surface, destroyed the integrity of the lysosomes, and induced the apoptosis and necrosis of cells. Nano-HAP crystals had a higher specific surface area, a smaller contact angle, a higher surface energy, and a lower Zeta potential than those of micro-HAP. Therefore, the abilities of HK-2 cells to adhere to and endocytose nano-HAP crystals were greater than their abilities to do the same for micro-HAP crystals. The order of the endocytosed crystals was as follows: HAP-40 nm > HAP-70 nm > HAP-1 μm > HAP-2 μm. The endocytosed HAP crystals entered the lysosomes. The more crystal endocytosis and adhesion there is, the more toxic it is to HK-2 cells. The results of this study showed that nanosized HAP crystals greatly promoted the formation of kidney stones than micrometer-sized HAP crystals.

The urinary tract is constantly exposed to microorganisms. Host defense mechanisms in protection from microbial colonization and development of urinary tract infections require better understanding to control kidney infection. Here we report that the lectin collectin 11 (CL-11), particularly kidney produced, has a pivotal role in host defense against uropathogen infection. CL-11 was found in mouse urine under normal and pathological conditions. Mice with global gene ablation of Colec11 had increased susceptibility to and severity of kidney and to an extent, bladder infection. Mice with kidney-specific Colec11 ablation exhibited a similar disease phenotype to that observed in global Colec11 deficient mice, indicating the importance of kidney produced CL-11 for protection against kidney and bladder infection. Conversely, intravesical or systemic administration of recombinant CL-11 reduced susceptibility to and severity of kidney and bladder infection. Mechanism analysis revealed that CL-11 can mediate several key innate defense mechanisms (agglutination, anti- adhesion, opsonophagocytosis), and limit local inflammatory responses to pathogens. Furthermore, CL-11-mediated innate defense mechanisms can act on clinically relevant microorganisms including multiple antibiotic resistant strains. CL-11 was detectable in eight of 24 urine samples from patients with urinary tract infections but not detectable in urine samples from ten healthy individuals. Thus, our findings demonstrate that CL-11 is a key factor of host defense mechanisms in kidney and bladder infection with therapeutic potential for human application.

Introduction: The relationship between trace elements and Oxidative Stress (OS) in Chronic Kidney Disease (CKD) patients is still not completely elucidated. The aim of this work is to determine the serum levels of OS and the trace elements in CKD patients.

Background Peritoneal fibrosis (PF) can reduce the efficiency of peritoneal dialysis and eventually lead to ultrafiltration failure. Epithelial-mesenchymal transition (EMT) of peritoneal mesothelial cells (PMCs) is the start of PF. Macrophages are involved in the process. This study was to investigate the effect of macrophage polarization on EMT of PMCs. Methods Monocyte-macrophage cells (THP-1) were treated to induce macrophage subsets (M1, M2a, M2c). The inducing was assessed by detecting protein and mRNA expression of cytokines using ELISA and RT-PCR. Subsequently, PMCs were co-cultured with M1, M2a and M2c, respectively, in Transwell chambers for 48 h and then expressions of E-cadherin and alpha-SMA were determined in PMCs. The PMCs that were not co-cultured with macrophages served as control PMCs. One-way ANOVA and SNK-q test were used to conduct statistics and P Results Detection of the cytokines, including IL-6, IL-10, IL-12, TGF-beta 1, CCL17 and CXCL13, verified that the inducting of macrophage subtypes was successful. Compared to control, E-cadherin protein expression was significantly decreased and alpha-SMA protein expression increased in M1-treated PMCs (P < .05); M2a-treated PMCs had an increased gene expression of alpha-SMA (P < .05); E-cadherin protein and gene expression were decreased and alpha-SMA protein and gene expression increased significantly in M2c-treated PMCs (P P < .01). Conclusions EMT of PMCs is enhanced by M2c macrophage polarization; meanwhile, M1 and M2a polarization may have the effect to some extent, but not as definite as M2c.

Background/Aims: The imbalance of T helper 17 (Th17) and regulatory T (Treg) cells exists in the occurrence and development of various diseases. Endoplasmic reticulum stress (ERS) is an important self-protective cellular response to harmful stimuli, such as uremic environment. The objective of this study was to investigate the Th17/Treg cell balance and ERS in a uremic environment and analyze the relationship between them. Methods: (1) The rat spleen lymphocytes were extracted and treated with thapsigargin (inducer of ERS) and sodium citrate. The proportion of Th17 and Treg cells were then detected. (2) The uremic serum-cultured lymphocytes were used and divided into three groups: non-uremic serum group, uremic serum group, and uremic serum + sodium citrate group. Afterward, the proportion of Th17/Treg cells and the expression of ERS-related proteins (GRP78 and CHOP) were detected. Results: Thapsigargin had no significant effect on the proportion of Th17 cells within a limited concentration range, but it could reduce the proportion of Treg cells, sodium citrate had a negative influence on the deviation of Th17/Treg cells treated with thapsigargin. Uremic serum treatment reduced the proportion of Treg cells, resulting in an increase of the Th17/Treg ratio. However, sodium citrate had no influence on the deviation of Th17/Treg cells treated by uremic serum. Sodium citrate reduced the elevation of ERS-related proteins induced by uremic serum. Conclusions: Uremic serum can lead to the imbalance of Th17/Treg cells as well as ERS, suggesting that ERS is one of the mechanisms of the imbalance of Th17/Treg cells induced by uremic serum. Sodium citrate can inhibit ERS induced by uremic serum.

目的 探究中药复方三参通脉合剂对颈动脉的粥样硬化斑块及血管内皮细胞代谢的潜在影响.方法 筛选2016年4月-2019年10月期间来该院内科门诊确诊颈动脉粥样硬化并自愿接受治疗的患者74例,其中治疗组38例,对照组36例.相较于单独给予阿司匹林的对照组,治疗组患者应用20 mL三参通脉合剂+阿司匹林的联合治疗方案,分别于治疗前后对所有患者颈动脉斑块的基本情况(数量、大小、平均厚度和颈动脉内-中膜厚度等指标)、血管内皮功能及其代谢产物水平进行检测.结果 治疗结束后,治疗组在斑块基本情况、血管内皮细胞代谢产物方面显著改善(P＜0.05).治疗组患者不仅显著增加NO水平,降低ET-1和NF-κB等内皮细胞代谢产物的表达,还能升高内皮依赖性血管舒张功能,一定程度改善内皮依赖性血管舒张功能(P＜0.05).结论 三参通脉合剂可缓解颈动脉粥样硬化症患者血脂参数,同时通过改善血管内皮细胞代谢功能,有效抑制和逆转病情进展.

[目的]探讨慢性肾炎患者尿N-乙酰-β-D-葡萄糖苷酶(NAG)、β2-微球蛋白(β2-MG)水平及临床意义.[方法]比较慢性肾炎患者和同期体检的健康成年人(对照组)尿NAG、β2-MG、血炎症因子超敏C反应蛋白(hs-CRP)、肿瘤坏死因子-α(TNF-α)、肾功能指标血肌酐(SCr)、尿素氮(BUN)和营养指标白蛋白(ALB)、血红蛋白(HGB)、红细胞计数(RBC)的差异,并分析慢性肾炎患者尿NAG、β2-MG与相关指标的相关性.[结果]慢性肾炎组患者的NAG、β2-MG水平高于对照组(P＜0.01);慢性肾炎组患者的hs-CRP、TNF-α和SCr、BUN水平高于时照组(P＜0.01),RBC、HGB和ALB水平均低于对照组(P＜0.01);慢性肾炎患者NAG、β2-MG与hs-CRP、TNF-a、SCr、BUN呈正相关,与RBC、HGB和ALB呈负相关.[结论]慢性肾炎患者NAG、β2-MG水平较高,且与炎症因子、肾功能等指标相关.

目的 探讨枸橼酸二乙酯(diethyl citrate,Et2 Cit)抑制慢性肾衰竭(chronic renal failure,CRF)血管钙化氧化应激的作用及其机制.方法 SD大鼠分4组:正常对照组、模型组、Et2Cit组,Et2Cit+NPS2143(钙敏感受体抑制剂)组;茜素红染色检测大鼠主动脉钙化水平;利用试剂盒检测大鼠主动脉和血浆中超氧化物歧化酶(superoxide dismutase,SOD)和一氧化氮(nitric oxide,NO)的含量.细胞分4组:正常对照组、高磷组、Et2 Cit组和Et2 Cit+NPS2143组;超氧化物阴离子荧光探针(dihydroethidium,DHE)染色半定量检测细胞DHE水平,流式细胞仪定量检测细胞DHE和活性氧(reactive oxygen species,ROS)水平.结果 与对照组相比,模型组主动脉钙化程度明显升高,伴血浆和主动脉SOD和NO水平降低(P<0.05);Et2Cit干预能降低主动脉钙化并增加SOD和NO水平(P<0.05);当同时给予NPS2143时,Et2 Cit抑制钙化的作用减轻,伴SOD和NO水平降低(P<0.05).高磷环境下细胞中DHE和ROS含量升高(P<0.05),Et2Cit干预能降低DHE和ROS含量(P<0.05);与Et2Cit干预组相比,当同时给予NPS2143时,细胞DHE和ROS水平升高.结论 Et2 Cit抑制CRF血管钙化的氧化应激水平,该作用依赖于钙敏感受体.

目的 探讨钙敏感受体(calcium sensing receptor,CaSR)/自噬信号轴在枸橼酸二乙酯(diethyl citrate,Et2 Cit)抑制慢性肾脏病(chronic kidney disease,CKD)血管钙化中的作用.方法 将大鼠和血管平滑肌细胞(vascular smooth muscle cells,VSMCs)分别分为正常对照组、模型组及低、高剂量Et2 Cit组和高剂量Et2 Cit+NPS-2143(CaSR抑制剂)组进行干预,试剂盒法检测各组大鼠主动脉钙含量;茜素红染色法检测各组细胞钙化情况;qRT-PCR法和Western blotting检测各组大鼠主动脉中钙化相关蛋白、CaSR和自噬相关蛋白的mRNA和蛋白表达.结果 与对照组相比,模型组钙化显著增加,Et2 Cit干预呈剂量依赖性地降低钙化程度(P<0.05);高剂量Et2 Cit+NPS-2143组钙含量较高剂量Et2 Cit组显著增加(P<0.05);与对照组相比,模型组平滑肌22α(smooth muscle 22α,SM22α)、CaSR和Beclin1的mRNA和蛋白表达降低(P<0.05),核心结合因子(runt related transcription factor 2,RUNX2)和P62表达升高(P<0.05);Et2 Cit干预能逆转以上变化(P<0.05);与高剂量Et2 Cit组相比,高剂量Et2 Cit+NPS-2143组的SM22α、CaSR和Beclin1显著降低,而RUNX2和P62显著增加(P<0.05).结论 Et2 Cit抑制CKD血管钙化,这一过程依赖CaSR和CaSR/自噬信号轴.

Vascular calcification (VC) is a common pathological manifestation in patients with cardiovascular diseases, leading to high mortality in patients with chronic kidney diseases. The deposition of hydroxyapatite (HAP) crystals on vascular smooth muscle cells leads to cell damage, which promotes osteogenic transformation. In this study, four different molecular weights (MWs ) of Porphyra yezoensis polysaccharides (PYP1, PYP2, PYP3, and PYP4 with MWs of 576, 49.5, 12.6, and 4.02 kDa, respectively) were used to coat HAP, and the differences in toxicity and calcification of HAP on A7R5 cells before and after coating were studied. The results showed that PYPs could effectively reduce HAP damage to the A7R5 cells. Under the protection of PYPs, cell viability increased and lactate dehydrogenase release, active oxygen level, and cell necrosis rate decreased; also, the amount of the HAP crystals adhering to cell surfaces and entering cells decreased. PYPs with low molecular weights presented better protective effects than high-molecular-weight PYPs. PYPs also inhibited the osteogenic transformation of the A7R5 cells induced by HAP and decreased alkaline phosphatase (ALP) activity and expressions of bone/chondrocyte phenotype genes (runt-related factor 2, ALP, osteopontin, and osteocalcin). In the adenine-induced chronic renal failure (CRF) mouse VC model, PYP4 was found to obviously inhibit the aortic calcium level, and it also inhibited the serum creatinine, serum phosphorus and serum BUN levels. PYP4 (least molecular weight) showed the best inhibitory effect on calcification and may be considered as a candidate drug with therapeutic potential for inhibiting cellular damage and osteoblast differentiation induced by the HAP crystals.

目的 探讨枸橼酸盐两段式抗凝在含钙置换液连续性静脉-静脉血液滤过(CVVH)中的抗凝效果及安全性.方法 将应用枸橼酸钠抗凝CVVH的79例患者随机分为两组.对照组:枸橼酸盐一段式抗凝39例,即4％枸橼酸钠溶液在滤器前以200ml/h的速度持续泵入;试验组:枸橼酸盐两段式抗凝40例,即以180ml/h速度在滤器前持续泵入4％枸橼酸钠溶液,同时在静脉壶泵入20 ml/h(滤器前与静脉壶处泵入的枸橼酸钠量比例9∶1).两组均使用含钙置换液(钙离子为1.5mmol/L),且不用在静脉回血端常规补充钙剂.观察两组滤器及静脉壶的凝血情况,根据凝血分级计算抗凝有效率.检测治疗前和治疗 2 h,6 h和12h血常规、凝血系列、血气分析、钙离子(Ca2+)浓度.结果 4％枸橼酸钠溶液输注总量在试验组为(200.42±5.85)ml/h,对照组为(202.05±8.33)ml/h,两组间比较差异元统计学意义(P =0.325).抗凝有效率在静脉壶部位试验组明显高于对照组(P=0.026),在滤器部位两组差异无统计学意义(P=0.986).试验组在 CVVH治疗过程中 2,6,12 h外周血及滤器后游离Ca2+浓度与对照组相比差异无统计学意义.结论 枸橼酸钠两段式抗凝与传统的一段式抗凝相比较,在含钙置换液连续性肾脏替代治疗中是安全的,且更为有效.

目的 探讨海藻异枝麒麟菜多糖(ESP)对纳米羟基磷灰石(nano-HAP)晶体诱导的小鼠血管平滑肌细胞(MOVAS)损伤的影响.方法 将体外培养的血管平滑肌细胞分为空白对照组、模型组、实验组.空白对照组加入新鲜培养基,模型组给nano-HAP刺激24 h,实验组用不同浓度(50、150、450 μg/mL)的ESP处理模型组细胞.采用MTT法检测细胞的活力,乳酸脱氢酶(LDH)试剂盒检测细胞LDH释放量,采用苏木精-伊红染色观察细胞形态.结果 与空白对照组比较,模型组细胞活力明显下降(P＜0.05),LDH释放量明显升高(P＜0.05);与模型组比较,实验组细胞活力随着加入ESP浓度的增加而升高,LDH释放量随着ESP浓度的增加而逐渐下降,差异均有统计学意义(均P＜ 0.05).结论 ESP可减轻nano-HAP晶体对血管平滑肌细胞膜的损伤.

提高住院医师的规范化培训质量是临床医学人才培养的关键,本文评价了微信学习互动群在肾病内科住院医师规范化培训中的积极作用.通过建立微信群,加入肾病内科规培的学员103人,定期在群中学习讨论.学员出科时通过考核和问卷调查评价微信群的作用,并与往年非微信群学员的成绩进行比较.发现参与微信群学习讨论的学员成绩高于非参与组差异有统计学意义,问卷调查结果规培学员及带教教师对微信学习群满意度高.我们得出结论:微信群是一种快捷、高效、实用的住院医师规范化培训和管理方式.

The acute phase reactant C-reactive protein (CRP) binds with high affinity to fibronectin (FN), but this binding occurs only at pH 6.5 or lower, and the binding is inhibited by calcium ions at physiological pH. Since CRP in the circulating blood exists in a calcium-binding form, the interaction between CRP and FN in vivo has been uncertain. CRP can undergo a conformational rearrangement in the absence of calcium or in the local microenvironment (e.g., acidic pH) of inflamed tissue to dissociate into monomeric CRP (mCRP). Therefore, we tested whether these discrepancies can be explained by the different isoforms and locations of CRP. Surface plasmon resonance and ELISA assays showed that mCRP binds with high affinity to FN, and the binding of mCRP to FN was unaffected by calcium or pH. Peptide competition assay, deletion mutant binding assay and protein docking analyse verified that the binding site of mCRP to FN is residues a.a.35-47. Furthermore, mCRP can significantly enhance the adhesion of monocytes to FN as well as upregulate the adhesion molecules expression on endothelial cell. Colocalization of mCRP with FN was observed in mice with DSS-induced colitis, whereas there was very little signal orcolocalization of CRP. These results provide in vitro and in vivo evidence that mCRP formed by local dissociation from circulating CRP is the major isoform that interacts with FN and regulates FN-mediated monocyte adhesion, which is involved in the pro-inflammatory process.

目的 探讨PBL联合CBL教学法应用于肾内科临床见习的教学效果.方法 研究对象选取在我院肾内科见习的64名学生,随机分为实验组和对照组,每组各32人,对照组采用传统的教学法即讲授法,实验组采用PBL联合CBL教学法,见习任务结束后进行随堂测试和问卷调查评估教学效果.结果 PBL联合CBL教学法组在学习成绩及教学方式满意度等方面显著高于传统教学法组,实验组学生考核成绩为(89.6±5.1)分,对照组学生(83.4±4.2)分,差异具有统计学意义(P＜0.05).结论 PBL联合CBL教学法相较传统教学法,能有效提高学生的学习成绩和主观能动性,有利于培养其临床思维能力,值得推广.

Purpose: Kidney cancer is a lethal type of malignancy with high mortality. The chemotherapeutic agents used for the treatment of kidney cancer have several adverse effects, there-fore, there is need to explore new molecules for the treatment of this disease. In the current study, the anticancer activity of plant-derived stilbenoid Mulberroside-A (MA) was evaluated against the A498 kidney cancer cell line and the underlying mechanism was explored. Methods: The A498 cell viability was determined by WST-1 assay. DAPI and AO/EB staining were employed for the detection of apoptosis. Matrigel and wound heal assay were used for cell invasion and migration study, respectively. Protein expression was checked by immunoblotting. Results: The results showed that MA could inhibit the growth of the A498 cells dose-dependently. The IC50 of 20 mu M was observed for MA against the kidney cancer A498 cells. The anticancer effects of MA against these cells were due to apoptotic cell death. Apoptosis was associated with alteration of the Bax/bcl-2 ratio. In addition, MA could also suppress the migration and invasion of the kidney cancer A498 cells by targeting EGFR signalling pathway. Conclusion: In conclusion, MA may prove beneficial in the treatment of kidney cancer.