METTL3/N6‐methyladenosine/ miR‐21‐5p promotes obstructive renal fibrosis by regulating inflammation through SPRY1/ERK/NF‐κB pathway activation

Renal fibrosis induced by urinary tract obstruction is a common clinical occurrence; however, effective treatment is lacking, and a deeper understanding of the mechanism of renal fibrosis is needed. Previous studies have revealed that miR‐21 impacts liver and lung fibrosis progression by activating the SPRY1/ERK/NF‐kB signalling pathway. However, whether miR‐21 mediates obstructive renal fibrosis through the same signalling pathway has not been determined. Additionally, studies have shown that N6‐methyladenosine (m⁶A) modification‐dependent primary microRNA (pri‐microRNA) processing is essential for maturation of microRNAs, but its role in the maturation of miR‐21 in obstructive renal fibrosis has not yet been investigated in detail. To address these issues, we employed a mouse model of unilateral ureteral obstruction (UUO) in which the left ureters were ligated for 3, 7 and 14 days to simulate the fibrotic process. In vitro, human renal proximal tubular epithelial (HK‐2) cells were transfected with plasmids containing the corresponding sequence of METTL3, miR‐21‐5p mimic or miR‐21‐5p inhibitor. We found that the levels of miR‐21‐5p and m⁶A modification in the UUO model groups increased significantly, and as predicted, the SPRY1/ERK/NF‐kB pathway was activated by miR‐21‐5p, confirming that miR‐21‐5p plays an important role in obstructive renal fibrosis by enhancing inflammation. METTL3 was found to play a major catalytic role in m⁶A modification in UUO mice and drove obstructive renal fibrosis development by promoting miR‐21‐5p maturation. Our research is the first to demonstrate the role of the METTL3‐m⁶A‐miR‐21‐5p‐SPRY1/ERK/NF‐kB axis in obstructive renal fibrosis and provides a deeper understanding of renal fibrosis.     

兔慢性肾功能衰竭模型的建立与评价

目的建立兔慢性肾功能衰竭模型,为干细胞移植治疗和相关研究奠定基础。方法普通级大耳白兔随机分为正常对照组和单侧输尿管结扎（unilateral ureteral obstruction,UUO）组。UUO组于输尿管结扎后2、4、6、8周进行血生化肾功能指标检测,并取肾组织观察肾脏病理学改变,通过SPECT动态观察肾小球滤过率的变化,采用免疫组织化学方法观察肾组织转化生长因子-β1（TGF-β1）的表达情况。结果①UUO组术后第2周,出现明显的血肌酐升高,尿素氮术后第8周开始升高（P〈0.01）。②UUO组术后第4周,肾脏组织出现了早期间质纤维化的病理改变,术后第8周肾小球开始出现硬化,间质纤维化明显,皮质明显变薄。术后第12周,肾小球硬化比例增加,肾小管玻璃样变性,间质纤维化进一步加重（P〈0.05）。③SPECT动态观察肾小球滤过率,UUO组第4周GFR值比正常对照组降低,到第8周时,GFR值进一步下降,结扎侧肾脏功能降低甚至丧失。④免疫组织化学染色显示,TGF-β1在术后第4、8、12周均明显增强,并且各时间点表达均有显著差异（P〈0.05）。结论单侧输尿管结扎法成功制作比较稳定的慢性肾功能不全模型,UUO后第8周符合肾脏间质纤维化模型标准。     

Mutations in VEGFA are associated with congenital left ventricular outflow tract obstruction

Left ventricular outflow tract obstruction (LVOTO) comprises a spectrum of stenotic lesions. Previous studies have shown that the vascular endothelial growth factor (VEGF) signaling system plays a critical role in cardiac cushion formation, vasculogenesis, and angiogenesis. We hypothesize that VEGFA may be a potential candidate gene associated with the spectrum of LVOTO lesions. However, it remains unclear whether the VEGFA gene is responsible for the development of LVOTO malformations. In this study, we identified three exon mutations in the VEGFA gene in three of 192 nonsyndromic LVOTO patients, and the overall mutation frequency was 1.6% (3/192). The c.454C>T (p.Arg152X) nonsense mutation and c.19_22dupGACA (p.Thr8ArgfsX78) internal tandem duplication mutation each introduced a premature stop codon and are predicted to produce a truncated VEGFA protein. The c.998G>A missense mutation changes a highly conserved arginine to a glutamine at residue 333 (p.Arg333Gln). These mutations were carried by some family members, and average penetrance was 33.3%. The present study suggests, for the first time to our knowledge, that VEGFA mutations may be associated with congenital LVOTO malformations. We provide evidence that LVOTO is likely oligogenic.     

复方嗜酸乳杆菌片联合乳果糖对糖尿病便秘患者脑肠肽水平的影响

目的探讨复方嗜酸乳杆菌片联合乳果糖对糖尿病便秘患者脑肠肽水平的影响。方法选择2018年1月-2019年8月在我院内科门诊治疗的糖尿病便秘患者92例,随机分为观察组和对照组各46例。两组均予建立良好生活习惯、饮食调整、适当运动、排便锻炼及药物控制血糖等基础治疗。对照组在此基础上加乳果糖口服液15 mL/次,2次/d,口服。观察组在对照组基础上加复方嗜酸乳杆菌片1.0 g/次,3次/d,温开水服用。两组均连用12周。观察两组治疗前后血清脑肠肽[一氧化氮(NO)、P物质(SP)和血管活性肠肽(VIP)]水平及便秘症状评分的变化,并比较其临床效果。结果治疗12周后,两组血清NO和VIP水平及Wexner便秘症状评分较前下降,血清SP水平较前上升(P0.05),且治疗后观察组变化幅度较对照组明显(P0.05)。观察组临床总有效率为95.7%,明显优于对照组的82.6%(χ~2=4.040,P=0.046)。结论复方嗜酸乳杆菌片联合乳果糖治疗糖尿病便秘患者的效果确切,其作用机制可能是通过调节脑肠肽分泌,减少NO和VIP分泌,促进SP分泌,从而提高结肠传输功能。     

A method for creating reversible ureteric obstruction in the primate

A technique has been developed for the establishment of a state of reversible, ureteric obstruction in the primate. Ten adult males had baseline ^99mTc-DTPA renogram studies. A randomly selected ureter was totally occluded and obstruction confirmed on renogram. The occlusion was reversed and subsequent renograms confirmed recovery of activity in the obstructed kidneys of the eight animals who survived the reversal procedure. Seven were alive on conclusion of the study. Prevention of ureteric strictures was achieved with an intra-ureteric silastic tube. Autopsies demonstrated patency of every previously occluded ureter. This is the first study to be reported in primates, and the second overall, in which complete ureteric obstruction and its successful reversal has been confirmed on renogram using this surgical method. The technique is suitable for the study of the effect of reversible ureteric obstruction on renal function.     

Surgery for hypertrophic cardiomyopathy

Hypertrophic cardiomyopathy (HCM) is a genetically determined cardiac disease characterised by otherwise unexplained myocardial hypertrophy of the left ventricle, and may result in left ventricular outflow tract obstruction. It is the most common cause of sudden cardiac death in young adults due to arrhythmias. Septal myectomy is a surgical treatment for HCM with moderate to severe outflow tract obstruction, and is indicated for patients with severe symptoms refractory to medical therapy. The surgical approach involves obtaining access to the interventricular septum via transaortic, transapical or transmitral approaches, and excising a portion of the hypertrophied myocardium to relieve the outflow tract obstruction. Large, contemporary series from centres experienced in septal myectomy patients have demonstrated a low early mortality of <2 %, excellent long-term survival that matches the general population, and durable relief of symptoms.     

聚葡萄糖治疗功能性便秘的临床疗效分析

目的:研究聚葡萄糖治疗功能性便秘的有效性和安全性.方法:采用自身对照试验方法,选择30例功能性便秘患者,给予口服聚葡萄糖冶疗1周,观察服药前后便秘症状及大便性状情况,并进行量化评分比较.结果:服用后1周后,显效者占36.6%,有效率达73.3%;停用后随访第7d,显效者占39.9%,有效率达76.7%.与治疗前比较,治...     

TNFRSF13C 在单侧输尿管梗阻大鼠肾组织中的表达及其意义

目的：检测单侧输尿管梗阻（UUO）大鼠肾组织中B 细胞激活因子受体（TNFRSF13C）的表达变化,探讨其在肾间质纤维化 病变中的作用。方法：采用UUO法建立肾间质纤维化大鼠模型,20只成年雄性大鼠,随机分为4组,分别于术后0、3、7、14 天处死 大鼠。取左侧梗阻肾脏进行Masson染色,拍照后,采用双盲法评定各组肾小管间质纤维化程度。提取肾组织中总RNA,用实时荧 光定量聚合酶链反应（RT-PCR）法检测各组肾组织中TNFRSF13C基因表达情况。Pearson 检测TNFRSF13C表达量与肾小管间质 纤维化程度的相关性。结果：随着梗阻时间的延长,肾组织中TNFRSF13C 的mRNA 表达量进行性升高,与肾间质纤维化病变程 度一致,两者呈显著正相关（r=0.915,P<0.01）。结论：TNFRSF13C可能在肾间质纤维化病程中起到了重要作用,并有望成为慢性 肾脏病的临床监测指标。     

兔肾积水模型的建立及SPECT和CT灌注成像

目的探索建立肾盂输尿管连接部梗阻所致肾积水的动物模型的可行性;探讨CT灌注成像对积水肾脏肾功能的评估价值。方法10周龄雄性新西兰兔50只随机分组,假手术组20只,分离左侧输尿管后直接关腹。模型组30只,选用腰大肌包埋输尿管造成左侧肾盂输尿管连接部梗阻。术前两组进行单光子发射计算机体层成像（SPECT）比较左肾功能,检验无差异后在术后3月分别行左肾SPECT、CT灌注,以病理检查为佐证,观察两组参数变化,进行CT灌注各项参数和GFR的统计学相关性分析。结果模型组建模成功达70%,呈慢性肾积水病理表现,左肾皮髓质CT灌注参数BF、BV、PS均下降,与相应GFR呈高度正相关。结论腰大肌包埋输尿管的模型制作方法具有可行性。CT灌注参数可作为肾功能状态的评定指标,具有一定的临床指导意义。     

波形蛋白在实验性肾间质纤维化上皮-间充质转分化中的表达及意义

目的明确肾脏纤维化中波形蛋白表达在上皮-间充质转分化观察中的意义。方法雄性SD大鼠12只,随机分成假手术组和模型组,模型组行单侧输尿管梗阻术。造模后14天处死大鼠,分别采用免疫组织化学染色和Western印迹法对梗阻侧肾组织波形蛋白和α平滑肌肌动蛋白作定性和定量检测。并通过体外实验用TGF-β1刺激诱导人近端小管上皮细胞株(HK-2)发生上皮-间充质转分化。采用间接免疫荧光法对E-钙粘蛋白和波形蛋白进行染色,采用激光共聚焦显微镜观察细胞形态改变,并用蛋白印迹法定量检测HK-2细胞波形蛋白和α平滑肌肌动蛋白表达水平。结果萎缩和扩张的肾小管上皮细胞出现波形蛋白和α平滑肌肌动蛋白表达,肾组织中波形蛋白和α平滑肌肌动蛋白表达量显著增加;正常的HK-2细胞,细胞形态为不规则圆形,TGF-β1刺激后细胞伸展成长梭形;细胞α平滑肌肌动蛋白表达水平显著增加,波形蛋白表达水平无显著变化。结论体内实验研究上皮-间充质转分化,将波形蛋白作为间充质细胞标志物具有较好的参考价值,而体外研究中,其标志作用尚存在争议。