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。     

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...     

CT 与X线诊断肠梗阻的临床价值探讨

目的：探讨腹部X 线与CT 扫描鉴别急性肠梗阻的准确性,为临床诊断提供参考。方法：选取2011年8 月-2013 年8 月我院 收治的66 例急性肠梗阻患者的临床资料进行回顾分析。所有患者均经手术活检或病理诊断证实为急性肠梗阻。术后患者均行腹 部X 线及CT扫描检查,对两种方法判断肠梗阻的发生、梗阻部位、类型及病因与手术病理结果进行比较,评价并分析两种检查 方法的准确率。结果：66 例肠梗阻患者中,X线检出率为89.39%,CT 检出率为95.45%。X 线诊断小肠梗阻准确率为72.10%,CT 为86.05%;X线诊断结肠梗阻准确率为69.57%,CT 为86.96%。X线诊断肿瘤准确率为69.57%,CT 为86.96%;X线诊断肠粘连 准确率为67.86%,CT 为82.14%;X 线诊断肠套叠准确率为60.00%,CT 为80.00%;X线诊断单纯性机械性肠梗阻准确率为78.72%,CT为82.98%;X线诊断绞窄性肠梗准确率为73.68%,CT 为78.95 %。CT 对肠梗阻部位、病因及类型的诊断准确率高于腹部X 线片,差异具有统计学意义(P<0.05)。结论：腹部X 线与CT 用于诊断急性肠梗阻具有较高的准确率,但CT 对于肠梗阻部位、梗阻类型及梗阻病因的诊断优于X线片。     

Effect of Beta-Adrenergic Stimulation on the Circulatory Lymphocyte Count: Studies in Normals and in Patients with Chronic Airflow Obstruction

In eight non-allergic patients with chronic airflow obstruction (CAO) and eight age and sex matched, healthy control subjects the circadian variation in circulatory lymphocyte count was studied in relation to serum Cortisol and urinary epinephrine levels. In addition, we investigated the effect of the beta-adrenergic agent terbutaline on the lymphocyte count in two ways: as a long-term effect after 8 days of oral slow-release terbutaline with constant diurnal and nocturnal serum levels in patients, and as a short-term effect by a constant rate infusion of 0.2μg/min over 4hr in normals. Both patients and controls showed similar circadian patterns of urinary epinephrine excretion and lymphocyte counts. Patients with CAO, however, had significantly lower epinephrine levels and significantly higher lymphocyte counts at all hours of observation (every 4 hr from 0800 to 0800 hr the next day), as compared with normal controls. After 8 days of slow-release terbutaline the lymphocyte count in the patient group decreased to levels not significantly different from that of normals. The circadian rhythm of the lymphocytes, however, persisted under terbutaline therapy. No correlation existed between the lymphocyte count modulating factor, serum Cortisol and the lymphocyte count over 24 hr. On placebo infusion in the control persons an increase of lymphocytes over 4hr occurred, as a consequence of circadian rhythmicity. On terbutaline infusion a significant increase of lymphocytes after 1hr was follwed by a decrease to levels significantly below those on the placebo day. The same pattern was found in the leucocyte count. From this study it is concluded that beta-adrenergic stimulation corrects the relative lymphocytosis to counts comparable with normals. Other coinciding factors must regulate, however, the circadian rhythmicity.     

慢传输型便秘患者肠道粪便菌群的变化特征

目的通过16S rDNA高通量测序技术,研究慢传输型便秘(slow transit constipation,STC)患者粪便菌群的变化特征。方法选取12例慢传输型便秘患者(STC组)及12例健康志愿者(对照组),留取粪便标本后,应用16S rDNA高通量测序技术对标本进行测序,并分析测序结果。结果 STC组与对照组人群肠道在菌群物种种类(Chao1指数和Sobs指数)上差异存在统计学意义(Z=-2.771,P=0.006;Z=-2.425,P=0.015),对照组人群肠道菌群物种种类明显高于STC组;两组在物种均一性(Shannon指数、Simpson指数)上差异无统计学意义(Z=-0.520,P=0.603;Z=-0.348,P=0.728)。在门水平,两组人群粪便菌群多样性差异无统计学意义。在属水平,Phascolarctobacterium(Z=-2.194,P=0.028)、Streptococcus(Z=-4.157,P0.001)、Eggerthella(Z=-2.427,P=0.015)和Actinomyces(Z=-2.310,P=0.021)在STC组人群肠道中相对丰度增加,而Faecalibacterium(Z=-2.887,P=0.004)、Roseburia(Z=-3.118,P=0.002)、Sphingomonas(Z=-4.157,P0.001)、Variovorax(Z=-2.732,P=0.006)、Holdemania(Z=-1.970,P=0.049)、Phyllobacterium(Z=-2.732,P=0.006)、Pseudomonas(Z=-2.439,P=0.015)、Acinetobacter(Z=-2.732,P=0.006)、Aquabacterium(Z=-2.134,P=0.033)和Novosphingobium(Z=-2.134,P=0.033)在STC组中相对丰度减少。结论 STC组与对照组人群肠道在菌群物种种类丰富度上差异存在统计学意义,而在菌群分布均一性上差异无统计学意义;在菌属水平,两组人群的粪便菌群构成也存在较大差异。     

超声引导下肝内胆管置管治疗肝内胆管结石并发梗阻的临床效果观察

目的:探究超声引导下肝内胆管置管治疗肝内胆管结石并发梗阻的临床效果和安全性。方法:选择2014年1月至2018年1月于我院接受治疗的98例肝内胆管结石并发梗阻患者为研究对象,将患者按照入院顺序统一编号后,根据随机数字表法进行分为实验组与对照组,每组各49例患者。对照组患者于常规X线引导下行肝内胆管置管治疗,实验组患者在超声引导下实施肝内胆管置管治疗,对比两组患者穿刺次数、手术时间、术后并发症的发生情况,并对两组患者随访3个月,比较其结石残余率及治疗效果。结果:(1)实验组患者穿刺次数及操作时间均显著少于对照组(P0.05);(2)实验组患者术后各类并发症发生率为4.08%,明显低于对照组(20.41%,P0.05);(3)对照组患者后3个月的结石残余率为14.29%(7/49),实验组为2.04%(1/49),显著低于对照组(P0.05);(4)术后3个月,实验组患者治疗总有效率为97.96%,明显高于对照组(81.63%,P0.05)。结论:与常规X线引导下行肝内胆管置管治疗相比,超声引导下肝内胆管置管在治疗肝内胆管结石并发梗阻中具有较更好的治疗效果和安全性。     

Increased cellular senescence in the murine and human stenotic kidney: Effect of mesenchymal stem cells

Cell stress may give rise to insuperable growth arrest, which is defined as cellular senescence. Stenotic kidney (STK) ischemia and injury induced by renal artery stenosis (RAS) may be associated with cellular senescence. Mesenchymal stem cells (MSCs) decrease some forms of STK injury, but their ability to reverse senescence in RAS remains unknown. We hypothesized that RAS evokes STK senescence, which would be ameliorated by MSCs. Mice were studied after 4 weeks of RAS, RAS treated with adipose tissue‐derived MSCs 2 weeks earlier, or sham. STK senescence‐associated β‐galactosidase (SA‐β‐Gal) activity was measured. Protein and gene expression was used to assess senescence and the senescence‐associated secretory phenotype (SASP), and staining for renal fibrosis, inflammation, and capillary density. In addition, senescence was assessed as p16+ and p21+ urinary exosomes in patients with renovascular hypertension (RVH) without or 3 months after autologous adipose tissue‐derived MSC delivery, and in healthy volunteers (HV). In RAS mice, STK SA‐β‐Gal activity increased, and senescence and SASP marker expression was markedly elevated. MSCs improved renal function, fibrosis, inflammation, and capillary density, and attenuated SA‐β‐Gal activity, but most senescence and SASP levels remained unchanged. Congruently, in human RVH, p21+ urinary exosomes were elevated compared to HV, and only slightly improved by MSC, whereas p16+ exosomes remained unchanged. Therefore, RAS triggers renal senescence in both mice and human subjects. MSCs decrease renal injury, but only partly mitigate renal senescence. These observations support exploration of targeted senolytic therapy in RAS.     

儿童功能性便秘肠道传输功能与肠道微生态的关系

目的采用不透光X线标志物测定结肠传输指数(TI)对功能性排便障碍进行分型,探讨各型功能性便秘患儿之间及其与健康儿童之间肠道菌群差异。方法选取功能性便秘儿童40例,进行结肠传输试验并进行分型,其中33例出口梗阻型(出口梗阻组),7例慢传输型(慢传输组)。留取便秘儿童新鲜粪便样本,同时收集20例正常儿童(对照组)粪便标本,存放于-80℃冰箱。采用16S rRNA测序技术鉴定菌群。结果菌群多样性分析:出口梗阻组、慢传输组与对照组粪便标本在丰富度上差异存在统计学意义(P0.01),出口梗阻组、慢传输组的肠道菌群丰度明显增高,3组在多样性上差异无统计学意义。门水平分析:出口梗阻型与慢传输型相比、出口梗阻型儿童与健康儿童相比,差异均无统计学意义;拟杆菌门丰度慢传输型儿童较健康儿童高,差异存在统计学意义(P0.05)。属水平分析:出口梗阻型与慢传输型相比,Subdoligranulum、Ruminococcus_2、Lachnospira、Collinsella数量在慢传输型儿童中均明显减少,差异存在统计学意义(均P0.05);毛螺菌属、Lachnoclostridium、Anaerostipes、Dorea、志贺菌属丰度慢传输型儿童较健康儿童低(均P0.05);Blautia、Anaerostipes、Lachnoclostridium丰度出口梗阻型儿童较健康儿童低(均P0.05);Alistipes、Parasutterella、Ruminococcus_2、Ruminococcaceae_UCG-005、Ruminococcaceae_UCG-002丰度出口梗阻型儿童较健康儿童高(均P0.05)。结论出口梗阻型与慢传输型相比,有部分菌种在慢传输型便秘儿童中明显减少,这需要进一步研究论证;便秘儿童肠道菌群与健康人群存在较大差异,改变肠道菌群可能成为治疗慢性功能性便秘的新策略。慢传输型便秘病例数较少,需增加样本量后进一步分析。     

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.