Íleo biliar como causa de dolor abdominal en el paciente anciano

Gallstone ileus is a rare and potentially serious complication of cholelithiasis. It is defined as a mechanical intestinal obstruction secondary to the presence of a gallstone in the intestinal luz. The most frequent cause is impaction of the calculus in the ileum after passing through a bilioenteric fístula. It has a high morbidity and mortality rate, mainly due to the difficulty and delay in its diagnosis. A retrospective study is presented of 4 cases of gallstone ileus treated between 2013 and 2017 in the Hospital Nuestra Señora del Prado. An analysis was performed on the clinical characteristics, diagnostic tests, and surgical treatment.     

胆红素自由基对大鼠肝细胞损伤作用的研究

用不同自由基源处理的胆红素与鼠肝细胞相互作用,结果表明：胆红素自由基可引起鼠肝细胞脂质过氧化,使总谷胱甘肽及ＧＳＳＧ水平明显下降,细胞损伤后,乳酸脱氢酶外漏,上述结果与自由基浓度正相关,而与其种类无关,牛血清白蛋白有明显的抑制作用。差示光谱表明：胆红素可能与细胞色素Ｐ－４５０快速形成络合物。根据以上结果,重点讨论了胆红素自由基对肝细胞损伤的化学本质及其与胆结石形成的关系。     

Hepatic Canalicular Membrane Transport of Bile Salt in C57L/J and AKR/J Mice: Implications for Cholesterol Gallstone Formation

C57L/J (gallstone-susceptible) and AKR/J (gallstone-resistant) mice have been utilized for quantitative trait loci (QTL) analysis to identify the Lith 1 locus for cholelithiasis. Abcb11 encodes for the liver canalicular membrane bile salt export pump (BSEP), which maps to this QTL and is a candidate gene for Lith 1. We investigated the transmembrane transport of taurocholate in canalicular liver membrane vesicles isolated from these murine strains. Canalicular liver plasma membranes (cLPM) and RNA were isolated from C57L/J and AKR/J mice livers, and were utilized for Northern and Western blot analysis and functional ³H-taurocholate uptake studies. ATP-dependent ³H-taurocholate uptake was significantly higher in AKR/J, compared to C57L/J mice. V _max was 127 vs. 42 pmol TC/mg/s in the murine strains, respectively, while K _m was unchanged. In contrast, gene and protein expression of hepatic Abcb11 was increased three-fold in C57L/J, compared to AKR/J mice. Thus, Abcb11 bile salt transport activity per unit protein was reduced nine-fold in the C57L/J, compared to AKR/J mice. In contrast, canalicular membrane cholesterol:phospholipid content was also significantly higher in the C57L/J mice. We conclude that gallstone-susceptible C57L/J mice demonstrate increased gene and canalicular membrane expression of Abcb11, however, taurocholate transport is functionally diminished. The latter may be due to the increased cholesterol membrane content of the cLPM in C57L/J mice. These findings may be important for the pathogenesis of gallstone formation.     

胆汁β-葡萄糖醛酸苷酶的性质及其计算荧光法

 胆汁经Sephadex G-25分子筛层析和DEAE-纤维素吸附处理,除去其中直接胆红素及胆汁酸盐,然后对其中的β-葡萄糖醛酸苷酶(GUSB)的性质进行了详细研究:该酶的最适温度为56℃;最适pH为4.5;以4mu Gr为底物测得Km为0.68mmol/L;直接胆红素是其竞争性抑制剂,其k_i=2.04×10~(-3)mmol/L;PI_1=6.0—6.2,PI_2=7.2—7.4,MW=280kD。根据竞争性抑制的米氏方程式设计建立了一个在直接胆红素并存的条件下,定量测定胆汁GUSB的计算荧光法,准确性为97.97±1.79％,重复性CV=1.2％,测定时间大大缩短。利用本法在pH4.5和7.0条件下测定了20例胆红素胆结石和17例胆固醇胆结石患者的胆汁GUSB的真实活性。结果指出:不论在pH4.5或7.0条件下,前组活性均显著高于后组(P<0.01)。证明胆汁中高GUSB活性与胆红素胆结石的形成有密切关系。     

胆囊细菌L型

用非高渗透压培养基培养法从常规细菌学检查阴性的慢性胆囊炎胆囊组织、胆汁及胆石中分离细菌Ｌ型,获得了８６．０２～１００％的检出率。胆囊检出的细菌Ｌ型绝大多数是稳定Ｌ型,在常规细菌学培养基或Ｌ型高渗透压琼脂培养基上不能形成可见的生长现象。沸水浴１０分钟可杀死胆囊或胆结石中的细菌Ｌ型,多种抗生素能抑制胆囊细菌Ｌ型的生长。细菌稳定Ｌ型在胆囊中如此广泛存在,可能是慢性胆囊炎与胆囊结石的重要病因。     

MALDI-TOF mass spectrometry screening of cholelithiasis risk markers in the gene of HNF1alpha

In recent years MALDI-TOF MS gained importance for high-throughput DNA analysis. In the present study this technique was used for the pathogenetic analysis of gallstone disease. The intestinal apical sodium-dependent bile acid transporter (ASBT) shows a genetic association with gallstone disease. ASBT has 3 binding sites in its 5'UTR for hepatocyte nuclear factor 1alpha (HNF1alpha). We hypothesized that genetic alterations in the HNF1alpha gene could influence ASBT expression. The gene HNF1alpha was sequenced in 46 Stuttgart random samples, composed of 16 controls and 30 gallstone patients. Subsequently, two independent cohorts (Stuttgart: 67 gallstones carriers, 109 controls, Leutkirch: 112 gallstone carriers, 99 controls) were screened by MALDI-TOF MS. The subjects were further divided by gender and weight. 24 known polymorphisms and two novel SNPs in the 3'UTR of HNF1alpha were detected (c.*220G>A and c.*1151G>A). After gender-specific sub-division of the pooled cohorts, 4 SNPs resulted in significant differences between male gallstone carriers and male controls (Stuttgart/Leutkirch: rs2255531 OR=2.78; p=0.006, rs1169288 OR=2.13; p=0.032, rs7310409 OR=2.34; p=0.025 and rs1169294 OR=2.13; p=0.031). Two novel variants in the 3'UTR of HNF1alpha were detected and four SNPs of HNF1alpha show a significant association to cholelithiasis in male gallstone patients. This article is part of a Special Section entitled: Understanding genome regulation and genetic diversity by mass spectrometry.     

Critical role of cholic acid for development of hypercholesterolemia and gallstones in diabetic mice

We studied bile acid and cholesterol metabolism in insulin-dependent diabetes utilizing genetically modified mice unable to synthesize cholic acid (Cyp8b1-/-). Diabetes was induced in Cyp8b1-/- and wild type animals (Cyp8b1+/+) by alloxan, and the mice were fed normal or cholesterol-enriched diet for 10 weeks. The serum levels of cholesterol were strongly increased in diabetic Cyp8b1+/+ mice fed cholesterol, while diabetic Cyp8b1-/- mice did not show any aberrations regardless of the diet. Diabetic cholesterol-fed Cyp8b1+/+ mice had much higher biliary cholesterol and cholesterol saturation index than all other groups, their bile contained a large number of cholesterol crystals, and their canalicular cholesterol transporter Abcg5/g8 mRNA levels were much higher. Cyp7a1 mRNA levels were similar in all diabetic mice but higher compared to non-diabetic animals. The results indicate a critical role for cholic acid for the development of hypercholesterolemia and gallstones in our animal model.     

Measurement of bile acid in serum and bile with arylamine-glass-bound 3alpha-hydroxysteroid dehydrogenase and diaphorase

3Alpha-hydroxysteroid dehydrogenase (3-HSD) from Pseudomonas testosteroni and diaphorase (lipoyl dehydrogenase) from Clostridium spp. have been immobilized individually onto arylamine glass beads through diazotization. A cost-effective enzymic colorimetric method for determination of bile acid in serum and bile employing a mixture of these immobilized enzymes was developed. The method is based on measurement of reduced nicotinamide adenine dinucleotide generated from bile acid in serum/bile by immobilized 3alpha-HSD with a color reagent consisting of nitro blue tetrazolium chloride salt, oxidized nicotinamide adenine dinucleotide, and immobilized lipoyl dehydrogenase in 0.065 M sodium phosphate buffer, pH 7.0. Analytical recovery of added bile acid (50 and 200 micromol/L) was 95.57 and 85.46% in serum and 97.6 and 91.6% in bile, respectively. Within- and between-batch coefficients of variation (CV) for bile acid determination were <1.2 and <0.2% in serum and >0.1 and <0.1% in bile, respectively. Good correlations for bile acid in serum (r1=0.92) and in bile (r2=0.97) were obtained by use of a standard chemical method and the present method. The mixture of immobilized 3alpha-HSD dehydrogenase and lipoyl dehydrogenase lost 50% of its initial activity after 6 months of regular use. The cost of bile acid determination in 100 serum and bile samples by the present method has been compared with that of the Sigma kit method.     

Lack of adiponectin promotes formation of cholesterol gallstones in mice

Plasma adiponectin levels are reduced in obese people, and hypoadiponectinemia is recently reported to associate with cholesterol gallstone formation in human. The aim of this study was to examine the role of adiponectin in gallstone formation using adiponectin knockout mice. We analyzed male knockout and C57BL6J mice fed normal or lithogenic diet for 6 weeks. On lithogenic diet, the prevalence rate of gallstone was significantly greater in knockout mice than C57BL6J mice. The molar percentages of β and ω-muricholic acid were significantly higher and hepatic sterol 12α-hydroxylase expression (cyp8b1) was significantly lower in knockout mice than C57BL6J mice fed normal diet. The bile apolipoprotein A-I protein levels were decreased in knockout mice. Histological examination showed gallbladder wall thickening and accumulation of glycoprotein in the gallbladder of knockout mice. Gallbladder phospholipase A2-IVA expression was significantly higher in knockout mice than in C57BL6J mice fed lithogenic diet. Our results indicate that lack of adiponectin promotes gallstone formation in mice.     

腹腔镜胆囊切除术治疗胆囊结石合并急性胆囊炎的疗效及对免疫功能和生活质量的影响

摘要 目的：观察腹腔镜胆囊切除术治疗胆囊结石合并急性胆囊炎的疗效及对免疫功能和生活质量的影响。方法：本次研究为回顾性研究,分析2018年3月~2021年3月期间我院收治的98例胆囊结石合并急性胆囊炎患者的临床资料,根据手术方案的不同将患者分为A组（n=46,给予开腹手术）和B组（n=52,给予腹腔镜胆囊切除术）,记录两组患者围术期相关指标、肝功能、免疫功能、生活质量和并发症发生率。结果：B组术中出血量少于A组,切口大小短于A组,手术时间长于A组,住院时间、首次排气时间短于A组（P<0.05）。两组术后3 d总胆红素 （TBIL）、谷丙转氨酶（ALT）、谷草转氨酶（AST）均升高,但B组低于A组（P<0.05）。B组术后3 d CD3⁺、CD4⁺、NK细胞、CD4⁺/CD8⁺高于A组（P<0.05）,B组术后3 d CD8⁺低于A组（P<0.05）。B组术后3个月健康生活量表简表（SF-36）各维度评分高于A组（P<0.05）。B组术后并发症发生率虽低于A组,但组间对比差异无统计学意义（P>0.05）。结论：腹腔镜胆囊切除术治疗胆囊结石合并急性胆囊炎,虽然手术时间较开腹手术更长,但切口小、可促进患者术后恢复,对患者免疫功能、肝功能损害更轻,有利于提高患者生活质量。