胆囊结石患者结石形态学特征及血浆脂多糖水平与急性胆源性胰腺炎的关系研究

摘要 目的：探讨胆囊结石患者结石形态学特征及血浆脂多糖（LPS）水平与急性胆源性胰腺炎（ABP）的关系。方法：选取2015年10月~2018年9月期间武汉大学人民医院收治的胆囊结石患者164例为研究对象,分析结石形态学特征与并发ABP的关系,同时采用Logistic回归分析ABP发生的危险因素。将所有患者根据LPS水平分为低LPS组（n=65,<10 pg/mL）以及高LPS组（n=99,≥10 pg/mL）,分析血浆LPS水平对不同结石大小、不同总胆固醇（TG）水平患者并发ABP的影响。结果：多发胆囊结石、球状结石、<3 mm结石、软碎型结石患者并发ABP的概率明显高于单发胆囊结石、不规则状结石或泥沙状结石、3~10 mm结石或>10 mm结石、硬型结石或胶冻型结石（P<0.05）。Logistic回归分析结果显示,多发胆囊结石、球状结石、<3 mm结石以及软碎型结石均是ABP发生的高危因素（P<0.05）。当患者处于高TG水平时,高LPS组并发ABP的概率高于低LPS组（P<0.05）,在细小结石患者中,高LPS组并发ABP的概率高于低LPS组（P<0.05）。结论：依据结石形态学特征可对胆囊结石患者并发ABP的可能性作出早期的判断,同时血浆LPS水平升高是高TG以及细小胆囊结石患者易并发ABP的重要因素之一。     

Functional Characterization of Na<Superscript>+</Superscript>/H<Superscript>+</Superscript> Exchangers in Primary Cultures of Prairie Dog Gallbladder

Gallbladder Na⁺ absorption is linked to gallstone formation in prairie dogs. We previously reported Na⁺/H⁺ exchanger (NHE1-3) expression in native gallbladder tissues. Here we report the functional characterization of NHE1, NHE2 and NHE3 in primary cultures of prairie dog gallbladder epithelial cells (GBECs). Immunohistochemical studies showed that GBECs grown to confluency are homogeneous epithelial cells of gastrointestinal origin. Electron microscopic analysis of GBECs demonstrated that the cells form polarized monolayers characterized by tight junctions and apical microvilli. GBECs grown on Snapwells exhibited polarity and developed transepithelial short-circuit current, I_sc, (11.6 ± 0.5 µA · cm^–2), potential differences, V_t (2.1 ± 0.2 mV), and resistance, R_t (169 ± 12 · cm²). NHE activity in GBECs assessed by measuring dimethylamiloride-inhibitable ²²Na⁺ uptake under a H⁺ gradient was the same whether grown on permeable Snapwells or plastic wells. The basal rate of ²²Na⁺ uptake was 21.4 ± 1.3 nmol · mg prot^–1 · min^–1, of which 9.5 ± 0.7 (~45%) was mediated through apically-restricted NHE. Selective inhibition with HOE-694 revealed that NHE1, NHE2 and NHE3 accounted for ~6%, ~66% and ~28% of GBECs total NHE activity, respectively. GBECs exhibited saturable NHE kinetics (V_max 9.2 ± 0.3 nmol · mg prot^–1 · min^–1; K_m 11.4 ± 1.4 mM Na⁺). Expression of NHE1, NHE2 and NHE3 mRNAs was confirmed by RT-PCR analysis. These results demonstrate that the primary cultures of GBECs exhibit Na⁺ transport characteristics similar to native gallbladder tissues, suggesting that these cells can be used as a tool for studying the mechanisms of gallbladder ion transport both under physiologic conditions and during gallstone formation.     

腹腔镜联合十二指肠镜治疗胆结石合并胆总管结石的临床疗效

目的:探讨腹腔镜联合十二指肠镜治疗胆结石合并胆总管结石的临床疗效。方法:收集我院肝胆科2012年6月到2014年3月住院的胆结石合并胆总管结石60例,按照随机数字表法分为试验组和对照组,各30例,试验组给予腹腔镜联合十二指肠镜治疗,对照组给予传统的开腹胆囊切除以及胆总管探查取石,观察对比两组的临床疗效。结果:试验组手术时间、住院时间、术后恢复时间及术中出血量明显的短于或少于对照组,差异均具有统计学意义(均P0.05),试验组治愈率为96.7%明显的高于对照组的66.7%,差异有统计学意义(P0.05)。两组主要不良反应有:结石残留、感染、十二指肠穿孔、恶性呕吐及伤口疼痛等,试验组不良反应发生率为16.67%明显的低于对照组总的73.33%,差异有统计学意义(P0.05)。结论:腹腔镜联合十二指肠镜治疗胆结石合并胆总管结石较传统的开腹手术治疗的效果好、恢复时间短、手术创伤小,且具有一定的安全性,不良反应发生少,值得临床推广使用。     

The genetic background of gallstone formation: An update

Gallstone disease is one of the most prevalent gastrointestinal diseases with a substantial burden to health care systems that is expected to increase in ageing populations at risk. This review summarizes recent data on the genetic background of cholesterol gallstones and the role of biliary lipid composition. Three previously unknown non-synonymous mutations in the ABCB4 gene encoding the hepatobiliary phospholipid-flippase MDR3 are presented.     

Calcium binding to biliary mucins is dependent on sodium ion concentration: relevance to cystic fibrosis

Because hypersecretion of gallbladder (GB) mucus occurs in gallstone formation and because binding of Ca(2+) to biliary lipids only accounts for 50% of the total Ca(2+) in GB bile, we investigated the binding of Ca(2+) to human biliary mucin. Biliary mucin was purified from GB bile and binding to Ca(2+) studied. Scatchard plot analysis suggested two binding sites. Removal of sialic acid by neuraminidase resulted in 10% reduction of Ca(2+) binding, whereas, sulfatase treatment reduced Ca(2+) binding by 30%. Using a hypotonic NaCl solution, Ca(2+) binding to mucin increased curvilinearly with mucin concentration. However, binding decreased with increasing ionic strength of the NaCl solution. We conclude that binding of Ca(2+) to mucin is effected mainly through sulfate. Binding to Ca(2+) can be displaced by Na(+). Ca(2+) binding to mucins is enhanced in the setting of low Na(+) concentrations. This phenomenon has pathophysiologic implications for the formation of thick mucus in cystic fibrosis epithelia.     

Characterization of Na+/H+ Exchanger Isoform (NHE1, NHE2 and NHE3) Expression in Prairie Dog Gallbladder

Gallbladder Na⁺ absorption is linked to gallstone formation in prairie dogs. Na⁺/H⁺ exchange (NHE) is one of the major Na⁺ absorptive pathways in gallbladder. In this study, we measured gallbladder Na⁺/H⁺ exchange and characterized the NHE isoforms expressed in prairie dogs. Na⁺/H⁺ exchange activity was assessed by measuring amiloride-inhibitable transepithelial Na⁺ flux and apical ²²Na⁺ uptake using dimethylamiloride (DMA). HOE-694 was used to determine NHE2 and NHE3 contributions. Basal J ^Na _ms was higher than J ^Na _sm with J ^Na _net absorption. Mucosal DMA inhibited transepithelial Na⁺ flux in a dose-dependent fashion, causing J ^Na _ms equal to J ^Na _sm and blocking J ^Na _net absorption at 100 μm. Basal ²²Na⁺ uptake rate was 10.9 ± 1.0 μmol · cm⁻²· hr⁻¹ which was inhibited by ∼43% by mucosal DMA and ∼30% by mucosal HOE-694 at 100 μm. RT-PCR and Northern blot analysis demonstrated expression of mRNAs encoding NHE1, NHE2 and NHE3 in the gallbladder. Expression of NHE1, NHE2 and NHE3 polypeptides was confirmed using isoform-specific anti-NHE antibodies. These data suggest that Na⁺/H⁺ exchange accounts for a substantial fraction of gallbladder apical Na⁺ entry and most of net Na⁺ absorption in prairie dogs. The NHE2 and NHE3 isoforms, but not NHE1, are involved in gallbladder apical Na⁺ uptake and transepithelial Na⁺ absorption. Received: 9 February 2001/Revised: 11 April 2001     

成石前后胆囊粘膜上皮多聚核蛋白体RNA含量变化—胆液糖蛋白和蛋白质增加的机制探讨

胆囊淤泥是胆囊结石的前身。测定胆囊粘膜上皮游离(FPR)和结合多聚核蛋白体(MBPR)的RNA含量及胆液粘液糖蛋白(MGP)的结果表明:胆囊淤泥患者胆囊粘膜合成和分泌MGP明显亢进,MBPR与相应GB中MGP的相关性比较提示此时尚有胆液淤积存在,且FPR的RNA含量也显著增高,提示细胞增殖加快。荧光胺法测定胆液蛋白质的含量发现,胆囊淤泥和胆固醇性结石患者胆囊液GB蛋白质含量显著高于色素性结石患者及“正常”对照,但蛋白质的GB浓度与HB浓度的比值显著低于胆固醇GB/HB比值,且GB中MGP与蛋白质含量呈显著正相关。说明蛋白质含量的增加与GB中过高的MGP阻止胆囊中蛋白质的清除过程有关。成石前GB中MGP和蛋白质含量的同时增高及其机制的确立,对认识及预防体内结石均有意义。     

胆结石应用腹腔镜与小切口手术切除胆囊治疗的临床效果分析

目的探讨腹腔镜与小切口手术切除胆囊治疗胆结石的临床疗效。方法选取2014~2015年我院收治的胆结石患者100例,随机分为对照组和观察组各50例,对照组患者采用小切口手术切除胆囊治疗,观察组患者行腹腔镜手术切除胆囊治疗,比较分析两组患者的临床效果。结果观察组术中出血量、手术时间、肠道功能恢复时间、住院时间及术后并发症发生率均有明显优势,与对照组比较差异均有统计学意义(均P0.05);观察组治疗总有效率为96%,对照组为84%,两组比较差异具有统计学意义(P0.05)结论采用腹腔镜胆囊切除术治疗胆结石患者,能明显提高临床疗效,值得临床推广应用。     

Preliminary study on the correlation between the trace Mn2+ and the calcite polymorph in gallstones containing calcium carbonate from the northeast China via electron spin resonance

Gallstones containing calcium carbonate (GCCC) from the northeast China were analyzed using X-ray diffraction (XRD), infrared spectroscopy (IR), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), and electron spin resonance (ESR). The sextet signal arising from the allowed transitions of the trace Mn²⁺ ions in GCCC was found to be ESR-detectable and strong. The XRD technique revealed the crystal habit of calcite in GCCC. Of the three polymorphs of calcium carbonate, no calcite was present as a solitary crystallization form, accompanied by aragonite or vaterite or both. The sextet ESR signal and the (104) main XRD peak at 2θ = ∼29.4° were employed as two probes to explore the relationship between trace Mn²⁺ and calcite. The Mn content can be considered as an indicator of the amount of calcite in GCCC because of the existence of a correlation between Mn²⁺ and calcite. The correlation between Mn²⁺ and calcite, the relation between the levels of Mn²⁺ and the type of gallstones, the structural preference of Mn²⁺ to the calcite polymorph, and the influence of dietary habits on calcite in calcium carbonate gallstones are discussed.     

ABCG5 and ABCG8 are expressed in gallbladder epithelial cells

Gallbladder epithelial cells (GBEC) are exposed to high biliary cholesterol concentrations on their apical (AP) surface. The mechanisms of cholesterol absorption and efflux by these cells are not known. We hypothesized that ABCG5 and ABCG8 are expressed in GBEC and mediate AP cholesterol efflux. Human gallbladder cDNA expressed message for ABCG5 and ABCG8. Cultured murine GBEC also expressed abcg5 and abcg8 mRNA and protein, as did cultured canine GBEC. Interestingly, treatment with model bile containing supersaturating concentrations of cholesterol, or treatment with LXRalpha/RXR ligands, did not lead to differences in expression of ABCG5 or ABCG8 in the murine or the canine cells. The subcellular localization of ABCG5 and ABCG8 did show alterations, with predominantly intracellular localization at baseline and predominantly AP localization following treatment with model bile or LXRalpha ligand. GBEC therefore express ABCG5 and ABCG8; these sterol transporters may play a role in mediating AP cholesterol efflux in the gallbladder epithelium.