Up-regulation of organic anion transporter 1 protein is induced by chronic furosemide or hydrochlorothiazide infusion in rat kidney.

BACKGROUND: Thiazide and loop diuretics are secreted from the proximal tubule via the organic anion transport system to reach their principal sites of action. Recently, a multispecific organic anion transporter 1 (OAT1) was identified in rat kidney and was localized to the basolateral membrane of the S2 segment in the proximal tubule. We postulated that interactions between thiazide or loop diuretics and OAT1 may play a role in the adaptation to long-term diuretic use, and investigated whether OAT1 is regulated in vivo by chronic administration of diuretics at the protein level. METHODS: Semi-quantitative immunoblotting and immunohistochemistry were carried out in kidneys from male Sprague-Dawley rats using a polyclonal peptide-derived antibody to OAT1. Furosemide (12 mg/day/rat, n = 6), hydrochlorothiazide (3.75 mg/day/rat, n = 6) or vehicle (1.7% ethanolamine, n = 6) were infused subcutaneously for 7 days using osmotic minipumps. Experimental and vehicle-control rats were pair-fed, and two bottles of drinking water were provided, one containing tap water and the other containing a solution of 0.8% NaCl with 0.1% KCl. RESULTS: Overt diuretic responses were observed to both furosemide and hydrochlorothiazide infusions. There were no differences in body weight or creatinine clearance between the experimental and control rats. Although OAT1 protein abundance in cortical homogenates was increased by furosemide infusion (271 +/- 35 vs 100 +/- 15%, P < 0.05), Na-K-ATPase alpha1 subunit protein abundance was not affected (113 +/- 14 vs 100 +/- 8%, P = 0.42). Immunohistochemical localization in tissue sections confirmed a strong increase in OAT1 expression in the basolateral membrane of the S2 segment of proximal tubule. OAT1 protein abundance in cortical homogenates was also increased by hydrochlorothiazide infusion (181 +/- 25 vs 100 +/- 7%, P < 0.01), whereas Na-K-ATPase alpha1 subunit protein abundance was not affected (105 +/- 4 vs 100 +/- 4%, P = 0.34). CONCLUSION: Chronic furosemide or hydrochlorothiazide infusion caused increases in OAT1 protein abundance in rat kidney. These results suggest that OAT1 may be up-regulated in vivo by substrate stimulation at the protein level.     

人组织激肽释放酶6在前列腺癌中的表达和意义

目的初步探讨人组织激肽释放酶6(Kallikrein 6,KLK6)在前列腺癌中的表达和意义。方法搜集徐州医学附属医院泌尿外科2010-06—2013-10行前列腺根治手术患者组织标本共65例,其中40例前列腺癌(Prostate Cancer,PCa),25例为前列腺增生组织,应用免疫组织化学两步法,检测组织中KLK6蛋白表达情况。结果 PCa患者组织中KLK6阳性表达率为75%,前列腺增生(Hyperplasia of prostate,BPH)组织KLK6阳性表达率为24%,两者比较癌组织显著高于增生组织(P0.01);前列腺癌中低危患者KLK6阳性表达率为58.25%,中高危患者为75%,两者比较中高危患者明显高于低危患者(P0.05)。结论 KLK6在前列腺癌组织表达明显高于前列腺增生,中高危患者明显高于低危患者,KLK6基因可能在前列腺癌诊断和预后判断中存在一定价值。     

肾上腺髓质增生的诊治(附9例报告)

目的探讨肾上腺髓质增生(AMH)的临床特点和诊治方法,提高AMH的诊治水平。方法回顾性总结9例AMH患者的临床资料,结合文献进行分析。结果9例患者均行手术治疗,5例术后血压明显下降,另4例血压恢复正常。9例术后病理均证实为AMH。结论影像学检查有助于AMH与嗜铬细胞瘤的鉴别诊断,确诊尚需术中探查和术后病理诊断。AMH根本治疗是手术切除,腹腔镜技术对AMH的手术治疗有广阔的应用前景。     

Urinary kallikrein excretion is related to renal function change and inflammatory status in chronic kidney disease patients receiving angiotensin II receptor blocker treatment

Aim: This study was to evaluate the correlation of urinary kallikrein to renal function, proteinuria and urinary cytokines in chronic kidney disease patients in a longitudinal follow up. Method: We measured urinary kallikrein and cytokines in 50 patients who were followed up for 12 months. Results: Using regression model we found that the kallikrein excretion (estimated by log kallikrein/creatinine) was positively correlated to log estimated glomerular filtration rate in the beginning and the end of follow up (P = 0.049 and 0.006, respectively). No correlation existed between kallikrein excretion and proteinuria. The kallikrein excretion decreased after 12 months of follow up, which was also associated with the decrease of log estimated glomerular filtration rate. There was a significant positive correlation between the log urinary kallikrein and monocyte chemoattractant protein‐1 (MCP‐1) concentration (correlation coefficient = 0.277; P = 0.049). Urinary kallikrein excretion was also positively correlated with serum MCP‐1 level (correlation coefficient = 0.431; P = 0.002). No correlation existed between urinary kallikrein and transforming growth factor β‐1 or tumour necrosis factor‐α concentration. Conclusion: Urinary kallikrein excretion is positively correlated to renal function, serum and urinary inflammatory mediator MCP‐1 in chronic kidney disease patients. These findings indicate that urinary kallikrein excretion may reflect the change of renal function and kidney inflammatory status.     

组织激肽释放酶对大鼠阴茎海绵体平滑肌cAMP 和cGMP影响的研究

目的探讨组织激肽释放酶对大鼠阴茎海绵体平滑肌cAMP和cGMP的影响，以初步阐明其舒张阴茎海绵体平滑肌的机制。方法采用放射免疫法，测定组织激肽释放酶对培养的大鼠阴茎海绵体平滑肌(去内皮组)和阴茎海绵体平滑肌组织(未去内皮组)cAMP和cGMP的影响，并以硝普钠为阳性对照，以赋形剂为阴性对照。结果100mu组织激肽释放酶使阴茎海绵体平滑肌组织内cAMP和cGMP浓度分别增高2．13倍和2．54倍，而对培养的阴茎海绵体平滑肌cAMP和cGMP没有明显作用。结论组织激肽释放酶能内皮依赖性地提高阴茎海绵体平滑肌细胞内cAMP和cGMP的水平，发挥舒张阴茎海绵体平滑肌的效应。     

The comparison of the diuretic and natriuretic efficacy of continuous and bolus intravenous furosemide in patients with chronic kidney disease

Aim: To compare natriuretic, kaliuretic, diuretic and free water clearance efficacy of continuous versus bolus intravenous furosemide administration in patients with chronic renal insufficiency. Material and methods: In a prospective randomized cross‐over trial, 42 patients of chronic renal insufficiency were randomized to receive the same dose of intravenous furosemide as bolus and continuous infusion. The effects of bolus and intravenous administration of furosemide on the volume of urine, sodium and potassium excretion were assessed. Results: Mean age was 53.6 ± 14 years and 23 (55%) were male. The mean modification of diet in renal disease glomerular filtration rate was 20.5 ± 17 mL/min per 1.73 m². The urinary excretion of sodium in intravenous bolus and infusion was 98.1 ± 78 and 114.4 ± 100 mmol, respectively (P = 0.001). Total urinary volume following bolus and infusion of furosemide was 1064 ± 627 and 1170 ± 764 mL, respectively (0.001). The excretion of potassium was similar in bolus (15.8 ± 16.6) and infusion (14.3 ± 9) administration (P = 0.11). The fractional excretion of sodium was higher following infusion (16.63 ± 16.1) than bolus administration (12.87 ± 9) of furosemide (P = 0.016). Conclusion: Continuous intravenous infusion of furosemide has significantly better natriuretic and diuretic effect than bolus administration of the same dose of the drug in patients with advanced chronic renal insufficiency.     

Assessment of protein synthesis and secretion by rat seminiferous and epididymal tubules in vivo

In vivo microperifbsion and micropuncture were used to study tubule protein synthesis and proluminal secretion by the male reproductive tract in vivo . Somniferous and caput and cauda epididymal tubules were perihsed for 3 h. with [³⁵S]-methionine. Perifused interstitial fluid (IF), lumen fluid (LF), and tubule extract (TE) were collected. Proteins were separated by SDS-PAGE, and autoradiograms were developed.
Trichloroacetic acid precipitable proteins in each fluid were determined and a protein synthesis index (PSI) was calculated. PSI values demonstrated that the cauda epididymis synthesized less protein in vivo than did either seminiferous or caput tubules.
Seminiferous tubules synthesized and secreted into the tubule lumen a relatively constant panel of proteins. Epididymal tubules synthesized and secreted proteins in a region-specific manner. In the caput epididymis the most prominent secreted bands were consistent with the heavy and light chains of epididymal clusterin. In the cauda epididymis, the most prominent synthesized and secreted protein was a 25 kDa protein consistent with the protein D. The above approach to studying protein synthesis and secretion will allow direct study of the physiological and path physiological effects on this important epithelial hnction in vim     

A randomized trial of furosemide vs hydrochlorothiazide in patients with chronic renal failure and hypertension.

BACKGROUND: Loop diuretics are the drugs of choice for the treatment of hypertension in chronic renal failure patients. However, the adaptive changes in the distal nephron and the short half-life of these drugs may decrease their long-term efficacy. Thiazides are not believed to be efficient in advanced renal failure, but this is debated. METHODS: We compared the efficacy of long-acting furosemide (60 mg/day) and hydrochlorothiazide (25 mg/day) in a double-blind, randomized crossover trial in seven patients with severe renal failure and hypertension (seven men, 54+/-10 years old). The primary end-points were sodium and chloride fractional excretions after 1 month of each diuretic and then after their combination. During the trial, other treatments and the diet were controlled. RESULTS: A trend towards an increase in the fractional excretion of sodium and of chloride was observed with furosemide, but the difference did not reach the level of statistical significance (P = NS). Hydrochlorothiazide significantly increased fractional excretion of sodium and chloride from 3.7+/-0.9 to 5.5+/-0.3 and from 3.9+/-0.19 to 6.5+/-0.3, respectively (P<0.05). The combination of the two diuretics had no additional effect on the increase in sodium and chloride fractional excretion. Furosemide, hydrochlorothiazide and the combination of the two diuretics decreased mean arterial blood pressure by the same extent from 112 to 97, 99 and 97 mmHg, respectively (P<0.05). CONCLUSIONS: Hydrochlorothiazide increased the fractional excretion of sodium and chloride more than furosemide did in hypertensive severe renal failure patients. Mean arterial blood pressure decreased by the same amount with both diuretics. Combining furosemide and hydrochlorothiazide did not increase the efficacy of hydrochlorothiazide.     

尿道电切镜外鞘联合输尿管镜治疗膀胱结石(附68例报告)

目的 探讨尿道电切镜外鞘联合输尿管镜钬激光治疗膀胱结石的优点与疗效.方法 68例膀胱结石患者,先置入Wolf F24尿道电切镜观察膀胱内部情况,退出内鞘及镜体,留置外鞘,F8输尿管镜由外鞘进入膀胱,钬激光碎石.结果 68例均一次手术成功,术后无结石残留,碎石率100％,手术时间20 ～ 60 min,平均(36.7±...     

Clinical significance of bradykinin liberation during cardiopulmonary bypass and its prevention by a kallikrein inhibitor

Activation of the kinin system and effects of Trasylol (Bayer, A.G. Lebukusen, West Germany), a kallikrein inhibitor, were investigated on 52 patients during hemodilutional cardiopulmonary bypass (CPB). Immediately after the start of CPB, neither elevation of bradykinin nor reduction of plasma kininogen (KGN: a precursor of bradykinin) were observed. During CPB, bradykinin level in the blood was markedly elevated, correlating with the significant decrease of kininogen (p<0.001). The longer the CPB time, the more marked the reduction of KGN. In the cases requiring over 60 minutes of CPB, the amounts of bradykinin released (4.6–18.0ng/ml) were sufficient to increase capillary permeability as well as peripheral vasodilatation. As shown by the significant increase of hematocrit (p<0.005) and the extreme reduction of vascular resistance found at the end of CPB in the prolonged cases. Infusion of Trasylol into the extracorporeal circuit actually prevented the reduction of kininogen and the increase of hematocrit as well as the extreme decrease of vascular resistance in the cases of over 60 minutes CPB. These results clearly point out that Trasylol is beneficial for the prevention of bradykinin liberation and capillary permeability increase and for the maintenance of optimum peripheral vascular tone during CPB. Furthermore, the significance of these findings with regards to complications during and after prolonged CPB was discussed.     

Dramatic suppression of plasma and urinary prostate specific antigen and human glandular kallikrein by antiandrogens in male-to-female transsexuals

PURPOSE: Prostate specific antigen (PSA) and human glandular kallikrein (hK2) are mainly produced by the prostate and their genes are regulated by androgens through the androgen receptor. We determine whether PSA and hK2 change significantly in plasma and urine after antiandrogen treatment in male-to-female transsexuals. MATERIALS AND METHODS: Plasma and urine PSA and hK2 were measured with highly sensitive immunofluorometric procedures capable of detecting within 1 or 6 ng./l. PSA or hK2, respectively. Study groups consisted of 10 men treated with cyproterone acetate only (group 1), 15 transdermal estradiol plus cyproterone acetate (group 2) and 31 ethinyl estradiol plus cyproterone acetate (group 3). Plasma and urine samples were collected before initiation of treatment as well as after 4 months of hormonal therapy. For a subset of group 3 patients blood and urine samples were also obtained after 12 months of treatment. RESULTS: Cyproterone acetate, a steroidal antiandrogen, alone or with estradiol was able to suppress greater than 90% of plasma and urinary PSA and hK2 concentration after 4 or 12 months of therapy. CONCLUSIONS: Cyproterone acetate therapy causes dramatic suppression of plasma and urinary PSA and hK2 in men without prostate cancer. Since cyproterone acetate is used for prostate cancer treatment, suppression of PSA after hormonal therapy may not accurately reflect therapy success in reducing tumor burden.     

利血平诱导的肾上腺髓质增生大鼠肾上腺髓质素的表达及其意义

目的观察肾上腺髓质素(ADM)在利血平诱导的大鼠肾上腺髓质增生动物模型的血浆和肾上腺髓质组织中的表达,探讨其意义。方法将雄性Wistar大鼠30只随机分为2组:A组(正常对照组)10只;B组(肾上腺髓质增生组)20只,免疫组织化学方法检测两组大鼠肾上腺组织中ADM蛋白的表达,同时检测血浆中ADM、儿茶酚胺(CA包括NE和E)、24 h尿3-甲基4-羟基苦仁酸(VMA)的浓度。结果6周末B组大鼠血浆CA以及24h尿VMA浓度均较A组大鼠高,并且在组织学上B组大鼠呈现增生改变;但两组之间的血压差异无统计学意义。B组肾上腺组织中ADM蛋白的表达明显高于A组(P<0.01);B组血浆中ADM的浓度与A组差异无统计学意义(P>0.05),且B组血浆中ADM与血浆中的NE、E和24 h尿VMA的浓度无明显相关(P>0.05)。结论肾上腺髓质可能不是血浆中ADM的主要来源;ADM在肾上腺髓质增生大鼠的血压调节中可能不起主要作用;组织中的ADM可能只是一个自分泌和旁分泌物质,在局部调节肾上腺细胞的功能。     

Identification of single nucleotide polymorphisms in the human kallikrein 10 (KLK10) gene and their association with prostate, breast, testicular, and ovarian cancers

BACKGROUND: The KLK10 gene (also known as the normal epithelial cell-specific 1 gene) is a member of the expanded human kallikrein gene family. Recently, it has been reported that KLK10 is a tumor suppressor gene and that its expression is downregulated in various forms of cancer and cancer cell lines. KLK10 is also upregulated in ovarian cancer. We thus hypothesized that the KLK10 gene may be a target for mutations in various cancers. METHODS: We sequenced the five coding exons of the KLK10 gene using genomic DNA from various tumors, normal tissues, and blood, by PCR amplification and automated sequencing. RESULTS: In none of the tumor-derived DNAs, we identified somatic mutations that could inactivate this gene. However, we identified a prevalent germline single nucleotide variation at codon 50 (exon 3) of this gene [GCC (alanine) to TCC (serine)]. The GCC genotype was less prevalent in prostatic cancer patients in comparison to control subjects (P = 0.027) but no differences were seen with testicular, ovarian, and breast cancer. We also identified four genetic variations in exon 4, at codons106 [GGC (glycine) to GGA (glycine)], codon 112 [ACG (threonine) to ACC (threonine)], codon 141 [CTA (leucine) to CTG (leucine)], and at codon 149 [CCG (proline) to CTG (leucine)]. None of these variations was significantly different between normal subjects and cancer groups. CONCLUSIONS: We found no evidence for somatic mutations of the KLK10 gene in cancers of the prostate, breast, ovary, and testis. The single nucleotide variation at codon 50 appears to be associated with prostate cancer risk.     

Cellular immunodeficiency in Paget's disease of bone: Changes induced by treatment with elcatonin

Summary We have found a cellular immune defect (low CD4 T lymphocyte count, high CD8 T lymphocyte count and a low CD4/CD8 ratio) in 39 PDB patients. This finding is not correlated with the bone metabolic activity of the disease. There was an improvement in the cellular immune defect in fifteen patients after treatment with elcatonin. These changes could be related to the improvement in the metabolic derangement or else could be the consequence of an effect on altered immunity.     

局部麻醉下外套式剥脱导管联合点状剥脱术治疗原发性大隐静脉曲张的效果分析

目的 探讨局部麻醉下外套式剥脱导管联合点状剥脱术治疗原发性大隐静脉曲张的效果.方法 选取2019年1月至2021年1月山东大学第二医院收治的482例原发性大隐静脉曲张患者,按照随机数字表法将其随机分为观察组(n=220)和对照组(n=262),观察组患者采用局部麻醉下外套式剥脱导管联合点状剥脱术,对照组患者采用神经阻滞...     

Continuous infusion vs. intermittent bolus injection of furosemide in acute decompensated heart failure: systematic review and meta‐analysis of randomised controlled trials

Loop diuretics remain a fundamental pharmacological therapy to remove excess fluid and improve symptom control in acute decompensated heart failure. Several recent randomised controlled trials have examined the clinical benefit of continuous vs. bolus furosemide in acute decompensated heart failure, but have reported conflicting findings. The aim of this review was to compare the effects of continuous and bolus furosemide with regard to mortality, length of hospital stay and its efficacy profile in acute decompensated heart failure. All parallel‐arm randomised controlled trials from MEDLINE, EMBASE, PubMed and the Cochrane Database of Systematic Reviews from inception until May 2017 were included. Cross‐over randomised controlled trials, observational studies, case reports, case series and non‐systematic reviews that involved children were excluded. Eight trials (n = 669) were eligible for inclusion. There was no difference between furosemide continuous infusion and bolus administration for all‐cause mortality (four studies; n = 491; I² = 0%; OR 1.65; 95%CI 0.93–2.91; p = 0.08) or duration of hospitalisation (six studies; n = 576; I² = 71%; mean difference 0.27; 95%CI ?1.35 to 1.89 days; p = 0.74). Continuous infusion of intravenous furosemide was associated with increased weight reduction (five studies; n = 516; I² = 0%; mean difference 0.70; 95%CI 0.12–1.28 kg; p = 0.02); increased total urine output in 24 h (four studies; n = 390; I² = 33%; mean difference 461.5; 95%CI 133.7–789.4 ml; p < 0.01); and reduced brain natriuretic peptide (two studies; n = 390; I² = 0%; mean difference 399.5; 95%CI 152.7–646.3 ng.l^?1; p < 0.01), compared with the bolus group. There was no difference in the incidence of raised creatinine and hypokalaemia between the two groups. In summary, there was no difference between continuous infusion and bolus of furosemide for all‐cause mortality, length of hospital stay and electrolyte disturbance, but continuous infusion was superior to bolus administration with regard to diuretic effect and reduction in brain natriuretic peptide.     

Acute effects of low and high intravenous doses of furosemide on myocardial function in anuric haemodialysis patients: a tissue Doppler study.

BACKGROUND: In patients with pulmonary oedema and preserved renal function, furosemide has not only a renal, but also a vascular effect, causing a rapid fall in left ventricular filling pressure accompanied by an increase in venous compliance. Previous studies have shown conflicting findings regarding the vascular effects of furosemide in patients with end-stage renal disease (ESRD). The objective of our study was to investigate whether furosemide induces changes in central cardiac haemodynamics in anuric ESRD patients, using conventional echocardiography and colour tissue Doppler velocity imaging (TVI), a new quantitative and sensitive method. METHODS: Repeated low doses (40 mg followed by an additional dose of 40 mg after 30 min) of i.v. furosemide were administered to 12 (61.6 +/- 16 years, 7 men) and a high dose (250 mg) of i.v. furosemide to 6 (64.1 +/- 3.6 years, 5 men) clinically stable anuric haemodialysis (HD) patients. Conventional two-dimensional echocardiography and colour TVI images were recorded immediately before (0 min) the furosemide infusion in both groups, and in the group receiving the repeated low-dose infusion (at 0 and 30 min), 10, 20, 30, 40, 50 and 70 min after the administration of the first infusion. In the group receiving the single high dose of furosemide the ultrasound investigation was repeated 10, 20, 30 and 40 min after the infusion. The myocardial tissue velocities (v; cm/s) for isovolumetric contraction (IVC), peak systole (PS), early (E') and late (A') myocardial diastolic filling velocities were measured in the left ventricle (LV) at six sites (infero-septal, antero-lateral, inferior, anterior, infero-lateral and antero-septal walls) at the basal region. IVC time (IVCT), IV relaxation time (IVRT), PS time (PSt), RR interval, mitral annulus motion (MAM), strain rate (SR), left ventricular filling pressure (E/E') and cardiac output were also measured. The average of the different walls was used to evaluate global function. Right ventricle (RV) dynamics was evaluated from measurements of IVC velocity (IVCv), peak systolic velocity (PSv), E' and A' from the RV free wall. RESULTS: No significant changes in cardiac output, IVCv, PSv, SR, MAM, E', A', E'/A', IVRT and LV filling pressure were observed, indicating that neither 40 mg (plus additional 40 mg after 30 min) nor 250 mg of furosemide had any measurable effects on LV filling pressure and LV and RV systolic and diastolic function. CONCLUSIONS: In anuric HD patients, low and high doses of furosemide had no significant effects on central cardiac haemodynamics. Therefore, the use of furosemide infusion in anuric ESRD patients with acute pulmonary oedema is not supported by the results of this study.