利用胆固醇氧化酶转化胆固醇制备胆甾-4-烯-3-酮

用酶法催化氧化胆固醇制备胆甾-4-烯-3-酮相对于化学法合成具有反应简单、成本较低等优点.作者探讨了在正辛烷作为有机相的两相反应体系中,以胆固醇氧化酶催化氧化胆固醇制备胆甾-4-烯-3-酮的方法.在胆固醇质量浓度为40g/L,反应时间40min、反应温度40℃、磷酸缓冲液-正辛烷体积比32、通氧40L/h及搅拌转速300r/min下,胆固醇转化率达到92%,经薄板层析及紫外扫描,发现转化产物为单一的胆甾-4-烯-3-酮.     

胆囊胆固醇结晶形成与载脂蛋白E相关性的研究

目的　探讨胆囊胆固醇结晶形成与载脂蛋白E(ApoE)的关系。方法　应用聚合酶链反应 (PCR)技术检测 80例胆囊胆固醇结石患者的ApoE基因多态性 ;采用偏振光显微镜观察 80例胆囊胆固醇结石患者胆汁中胆固醇单水结晶的形成及数目。结果　 80例胆囊胆固醇结石患者中有E4型 2 3例 ,其中 15例术后立即观察到胆固醇单水结晶 ,而E3 型 49例中有 2 0例术后立即观察到结晶 ,E2 型 8例中有 3例立即观察到结晶 ,E4 型明显高于E3 及E2 型 (P <0 .0 1) ,且ApoE4 型胆固醇单水结晶数目 (130 0 /mm3)明显高于E3(5 2 0 /mm3)及E2 (5 0 0 /mm3)。E4 型成核时间 (2 .5d)比E3(5 .5d)和E2 (6 .0d)明显缩短 (P <0 .0 1)。结论　胆囊胆固醇结晶形成与ApoE相关 ,ApoE4 为胆囊胆固醇结晶形成的一种遗传易感性因子。     

酶法回收胆固醇-β-环糊精包合物中的胆固醇

霉菌淀粉酶能有效地水解制备低胆固醇蛋制品时所生成的副产物胆固醇－β－环糊精包合物中的α－１，４糖苷键．霉菌淀粉酶水解胆固醇－β－环糊精包合物的最佳条件为底物质量浓度１５ ｇ／ｄＬ，水解时间３ ｈ，水解温度５６ ℃，ｐＨ ５．２．采用酶法技术从２ ｋｇ 干基胆固醇－β－环糊精包合物中回收制得胆固醇质量分数为８９．５％ 的制品，胆固醇回收率达７２％ ．     

术后早期低总胆固醇血症在预测肝切除术后严重并发症中的价值

目的：探讨术后早期低总胆固醇血症在预测肝切除术后严重并发症中的价值。方法回顾性分析2009年至2011年间接受肝切除术的350例患者术后早期血总胆固醇水平与术后严重并发症之间的关系。结果受试者工作特征曲线提示术前总胆固醇/术后5d内总胆固醇谷值比（以下简称术前/术后总胆固醇比）对术后严重并发症有预测价值,其曲线下面积为0．84（95％可信限0．78～0．90,P＝0．029）。依据该曲线,术前/术后总胆固醇比的诊断值为1．69;其敏感性为73．4％,特异性为85．2％。多因素分析显示术前/术后总胆固醇比＞1．69为术后严重并发症的独立危险因素。结论术后早期低总胆固醇血症对肝切除术后严重并发症有较好预测价值。     

小鼠胆囊胆固醇结石模型的建立

目的建立简单、可靠、高效的小鼠胆囊胆固醇结石模型,为研究胆石成因及防治提供重要手段。方法C57BL／6小鼠随机分为对照组和模型组,对照组喂饲基础饲料,模型组喂饲致石饲料（基础饲料加10％猪油、1％胆固醇及0．5％胆酸）。两组小鼠分别于喂养4周和8周后,计算小鼠存活率,同时各取一半数量小鼠在乙醚吸入麻醉下手术取胆囊及血液标本,分别检测成石率、血脂浓度及胆汁胆固醇饱和度。结果对照组小鼠8周存活率100％,模型组小鼠死亡1只,存活率95％。对照组8周成石率为零,模型组4周成石率80％,8周成石率100％。血脂分析表明,与对照组比较,模型组4周和8周总胆固醇及低密度脂蛋白显著升高（P〈0．01）,甘油三酯浓度轻度升高（P〈0．05）,高密度脂蛋白显著降低（P〈0．01）。4周和8周时胆汁胆固醇饱和度测定,对照组分别为0．48±0．29和0．58±0．21,模型组分别为1．36±0．36和1．52±0．37,模型组胆固醇浓度处于过饱和状态。结论本模型方法简单、成石率高、动物死亡率低,可作为研究胆石成因及防治的备选模型。     

胆固醇氧化酶转化蛋黄胆固醇工艺的优化

研究了用响应面分析法(RSM)优化酶法转化蛋黄胆固醇的工艺,发现胆固醇氧化酶(COD)添加量及蛋黄粉的稀释率是影响蛋黄胆固醇转化率最重要的因素.在模拟优化的条件下反应时间为14.15h,蛋黄粉稀释率为3.54,COD为5.39U/g时,胆固醇转化率达85.61%.对氧化产物进行分析发现仅有单一产物胆甾烯酮,该产物具有降血脂、减肥等功效.     

胆囊胆固醇息肉与胆固醇结石的胆汁成分比较

目的分析胆囊胆固醇结石与胆固醇息肉病人的胆囊胆汁成分,探讨胆固醇结石和胆固醇息肉形成中各种成分的差异。方法测定20例胆囊胆固醇息肉（息肉组）、20例胆囊胆固醇结石（结石组）和10例非肝胆疾病（对照组）的胆囊胆汁成分。采用HITACHI-7060全自动生化分析仪测定总胆汁酸（TBA）、磷脂（PL）和总胆固醇（TC）;氨基己糖比色法测定糖蛋白;ORION-720A型数字离子酸度计测定游离Ca^2＋浓度及pH值;Agilent-1100高效液相色谱分析仪测定8种结合胆汁酸的含量。结果息肉组和结石组TC[（14.0±0.5）mmol/L,（18.6±1.2）mmol/L]、CSI（1.217±0.039,1.565±0.087）、三羟/二羟胆汁酸比值（0.702±0.084,0.763±0.060）均显著高于对照组TC（9.1±0.8）mmol/L、CSI（0.812±0.075）、三羟/二羟胆汁酸比值（0.585±0.067）（P〈0.05）,而且上述指标结石组显著高于息肉组（P〈0.05）。2组TBA含量分别为（110.7±14.8）mmol/L、（105.8±16.5）mmol/L,显著低于对照组（137.6±33.1）mmol/L（P〈0.05）。结石组糖蛋白（1.8±0.2）mmol/L、游离Ca^2＋（2.2±0.3）mmol/L及pH值（7.9±0.3）显著高于息肉组和对照组糖蛋白（0.6±0.1）mmol/L,（0.7±0.1）mmol/L、游离Ca^2＋（1.2±0.2）mmol/L,（1.1±0.1）mmol/L、pH值（7.0±0.1）,（6.9±0.2）（P〈0.05）。结石组甘氨胆酸（G）/牛磺胆酸（T）比值（2.777±0.217）显著低于息肉组（3.624±0.465）和对照组（3.960±0.377）（P〈0.05）;结石组和息肉组PL/TBA（0.311±0.044,0.292±0.036）显著高于对照组（0.241±0.082）（P〈0.05）。结论胆囊胆汁中总胆汁酸含量的降低和总胆固醇浓度的升高,是形成胆固醇结石和胆固醇息肉的共同因素。结合胆汁酸构成比例、pH值、糖蛋白及游离钙离子浓度在胆囊胆固醇息肉及结石的胆汁中存在差别。     

骨桥蛋白与钙离子的相互作用及其在不同胆汁体系中对胆固醇成核效应的影响

目的:探究骨桥蛋白与钙离子的相互作用及其在不同胆汁体系中对胆固醇成核效应的影响。方法:选择2016年7月—2018年9月在我院治疗的100例胆固醇结石患者,将其分为单纯胆囊结石组和原发性胆管结石组各50例,再选取50例正常人作为对照组。采用酶联免疫法检测胆固醇结石患者和正常人胆汁中骨桥蛋白与钙离子的浓度,并采用Holan法测定3组研究对象在不同浓度骨桥蛋白与钙离子中的胆固醇成核时间。结果:单纯胆囊结石组和原发性胆管结石组患者的骨桥蛋白浓度均低于对照组,而钙离子浓度高于对照组,差异有统计学意义(P<0.05)。单纯胆囊结石组和原发性胆管结石组不同浓度的骨桥蛋白及钙离子的胆固醇成核时间均少于对照组,差异有统计学意义(P<0.05)。与同组50μg/mL浓度的骨桥蛋白相比,骨桥蛋白100μg/mL浓度的胆固醇成核时间更长,而骨桥蛋白50μg/mL+钙离子10 mmol/L浓度的胆固醇成核时间更短,差异有统计学意义(P<0.05)。与同组100μg/mL浓度的骨桥蛋白相比,骨桥蛋白100μg/mL+钙离子10 mmol/L浓度的胆固醇成核时间更短,差异有统计学意义(P<0.05)。结论:骨桥蛋白在不同胆汁体系中对胆固醇成核效应发挥抑制作用,而钙离子发挥促进作用。骨桥蛋白延长钙离子的促成核时间,钙离子缩短骨桥蛋白的抑成核时间,二者互相影响。     

胆囊黏膜ABCG5和ABCG8与胆囊胆固醇息肉关系的研究

目的:研究胆囊黏膜ABCG5和ABCG8的基因表达与胆囊胆固醇息肉发病的关系.方法:收集63例因胆囊疾患而行腹腔镜胆囊切除术患者的胆石、胆汁及部分胆囊黏膜组织,其中胆囊胆固醇息肉32例,胆囊胆固醇结石18例,对照13例(胆囊腺瘤6例,非胆固醇胆囊结石7例),分别测定胆石胆固醇含量,胆汁胆固醇、胆汁酸、磷脂的浓度,实时定量PCR检测胆囊黏膜ABCG5和ABCG8的mRNA表达.结果:胆固醇息肉组胆汁胆固醇饱和指数较对照组明显增高(P<0.01),结石组胆囊黏膜ABCG5和ABCG8的表达量较息肉组和对照组显著增高,息肉组和对照组ABCG5和ABCG8的表达量差异无统计学意义(P>0.05).结论:胆囊黏膜ABCG5/ABCG8 mRNA的相对低水平表达,可能是导致胆囊胆固醇息肉发病的重要因素之一.     

胆汁胆固醇对胆囊收缩素受体表达的影响

目的　探讨胆汁胆固醇对胆囊收缩素受体 (CCK R)表达的影响。　方法　采用放射免疫分析法和受体放射配基结合法检测对照组 (n =2 5 )、高胆固醇组 (n =2 5 )、自然恢复组 (n =2 5 )及治疗组 (n =2 5 )豚鼠门静脉血CCK水平、胆囊CCK R的最大结合容量 (Bmax)和亲和力 (Kd) ,同时观察空腹胆囊体积 (FV)、胆囊胆汁量 (FB)和餐后胆囊体积 (RV)、胆囊胆汁量 (RB)及胆囊收缩率 (E)、胆汁胆固醇浓度的变化。　结果　与对照组比较 ,高胆固醇组豚鼠FV[(0 89± 0 2 6 )～ (1 34± 0 6 1)cm3 ]、FB[(0 6 8± 0 2 0 )～ (1 0 1± 0 4 3)cm3 ]、RV[(0 2 8± 0 0 8)～ (0 90± 0 5 3)cm3 ]、RB[(0 2 3± 0 0 6 )～(0 83± 0 32 )cm3 ]增大 ,E[(6 5 83± 7 32 ) %～ (47 2 2± 5 2 4 ) % ]下降 ,胆汁胆固醇浓度 [(0 4 4±0 11)～ (0 6 0± 0 13)mmol/L]升高 ,门静脉血CCK水平及CCK R的Kd无改变 ,而CCK R的Bmax[(6 0± 2 7)～ (32± 13)fmol/mg蛋白 ]下降 ;与自然恢复组比较 ,治疗组上述各项指标正常。　结论　胆汁中的高胆固醇通过下调胆囊CCK R表达而导致胆囊收缩功能障碍 ,降低胆汁高胆固醇浓度可以促进胆囊动力功能的恢复。     

Cholesteroloma of the breast: A 10 year retrospective review of 79 cases with radiology correlation

A cholesteroloma or cholesterol granuloma of the breast is an uncommon lesion representing an inflammatory/reactive process with unclear etiology. In this study, we reviewed our 10‐year experience with cholesteroloma of the breast with clinical, radiologic, and histopathological correlation. Seventy‐nine cases were selected. The mean patient age was 57.7 (range 25‐90) years old. Patients had hypercholesterolemia with mean blood cholesterol level of 201 mg/dL (P < 0.001). The mean body mass index (BMI) was 26.7 kg/m² (P = 0.1976). The indications for the breast biopsies were mass lesion on radiology (85.5%, n = 65) and microcalcifications (10.5%, n = 8). Of the 65 cases of the mass lesions, 52 presented as solid masses and 13 were cystic. On the diagnostic mammogram or ultrasound, 81.9% were BI‐RADS 4% and 6.9% were BI‐RADS 5. Macrocysts were the most common pathological finding associated with cholesteroloma suggesting the etiology of cholesteroloma may be the result of repair process from obstruction and rupture of the macrocysts. Six cases (9.2%) of cholesterolomas had persistent masses during follow‐up. The recognition of this lesion and radio‐pathological correlation can help us better understand this entity and distinguish it from its mimickers.     

Cholesterol granuloma of the kidney mimicking a tumour.

Introduction Cholesterol granulomas are lesions consisting of cholesterolcrystals and foreign body giant cells [1]. Lipid disturbancesare thought to play a role in their aetiology. They are rarehistological     

Optimal low‐density lipoprotein concentration for cardiac allograft vasculopathy prevention

Cardiac allograft vasculopathy (CAV) is a major risk factor influencing graft loss and patient survival following orthotopic heart transplant. Allograft vasculopathy is a multifactorial process, which includes both immunologic and non‐immunologic mechanisms. Given the non‐immunological risk factors for vasculopathy, particularly hyperlipidemia, it is intuitive that reducing a patient's LDL would help attenuate the disease process. Multiple studies have shown benefits with the use of statin therapy. However, current heart transplant guidelines do not give a specific recommendation as to what LDL goal should be achieved in this patient population. This study is a retrospective cohort analysis designed to determine the relative risk of developing cardiac allograft vasculopathy with respect to different LDL goals. Median LDL level of <100 mg/dL was shown to significantly reduce the risk of developing cardiac allograft vasculopathy. Twelve of 37 patients with an LDL ≥100 mg/dL (32.4%) developed CAV vs 25 of 157 patients (15.9%) with an LDL <100 mg/dL (P = .021). Furthermore, a delay in to time to cardiac allograft vasculopathy was seen when a median LDL concentration of <100 mg/dL was achieved. This benefit was not extended when a goal concentration of <70 mg/dL was targeted.     

Lipid Parameters Including Lp(a) in Hemodialysis Patients

Chronic hemodialysis (CHD) patients have a high incidence and prevalence of atherosclerotic disease which may be related to numerous atherosclerotic risk factors. Among them dyslipidemia plays a significant role. Elevated Lp(a) levels, which are strongly associated with atherosclerosis, have been reported recently in uremic patients. The aim of our study was the determination of the levels of lipid parameters including Lp(a) in 151 CHD patients (76 male) aged 57 (12-81) years, who were on hemodialysis for a mean of 44.3 (range 1 to 189) months. Eighty-four normal individuals age and sex matched were used as controls. The median serum Lp(a) concentration in hemodialysis patients was 13 mg/dL compared with 6.5 mg/dL in healthy controls, p<0.001 by distribution-free Mann-Whitney test. The prevalence of subjects with Lp(a) levels above 25 mg/dL was significantly higher in CHD patients compared to normal subjects (30% vs. 8%, p<0.001). Even if CHD patients were matched for fasting lipid levels, they showed Lp(a) levels significantly higher than controls. No significant correlation was found between Lp(a) levels and either the age of the patients or the duration of hemodialysis. The etiology of primary renal disease did not influence the Lp(a) levels.     

Effect of MCI-196 (colestilan) as a phosphate binder on hyperphosphataemia in aemodialysis patients: a double-blind, placebo-controlled, short-term trial.

BACKGROUND: MCI-196 (colestilan), an anion exchange resin, is widely used as an anti-hypercholesterolaemic drug in Japan. To evaluate the efficacy and safety of MCI-196 as a phosphate binder, a double-blind, randomized, placebo-controlled prospective trial was conducted in Japanese end-stage renal disease patients with hyperphosphataemia on intermittent haemodialysis treatment. METHODS: Phosphate binders were discontinued during a 2-week washout period. Subsequently, patients whose serum phosphorus levels were > or =6.5 mg/dl, but <10 mg/dl were eligible to enter the treatment protocol. Patients were randomized to either MCI-196 6 g/day or placebo for 2 weeks. The efficacy and safety of MCI-196 were assessed in 33 and 46 patients, respectively. RESULTS: Serum phosphorus in the placebo group increased by 0.84+/-0.95 mg/dl (mean+/-SD), while serum phosphorus in the MCI-196 group decreased by 0.55+/-1.23 mg/dl. The difference between the two groups was statistically significant (P = 0.002). A reduction of > or =1 mg/dl in serum phosphorus was observed in 43% in the MCI-196 group and 0% of patients in the placebo group (P = 0.0122). Calcium-phosphorus (Ca x P) product, intact parathyroid hormone (iPTH) and low-density lipoprotein (LDL)-cholesterol in the MCI-196 group also decreased significantly compared with the placebo group, while serum calcium was unchanged. Adverse reactions were observed in 51.7% of the MCI-196 group and 29.4% of the placebo group (P = 0.2186). The most frequent adverse reactions in the MCI-196 group were gastrointestinal symptoms and signs, including constipation. CONCLUSIONS: The present findings suggest that the short-term administration of MCI-196 is effective in decreasing serum phosphorus in haemodialysis patients. Its long-term efficacy needs to be evaluated.     

The association of lipid levels with mortality in patients on chronic peritoneal dialysis.

BACKGROUND: The role of traditional risk factors, including plasma lipids, in the pathogenesis of cardiovascular (CV) disease in chronic dialysis patients is unclear. Previous studies have suggested that lower serum total cholesterol (TC) is associated with higher mortality in patients on chronic haemodialysis (HD). Whether this relationship is specific to the HD population or is common to the uraemic state is unclear. The present study evaluated the association of serum TC and triglycerides with clinical outcomes in chronic peritoneal dialysis (PD) patients. METHODS: Data of 1053 PD patients from the United States Renal Data System (USRDS) prospective Dialysis Morbidity and Mortality Study Wave 2 were examined. Cox regression was used to evaluate the relationship between lipid levels and mortality. RESULTS: Patients with TC levels < or =125 mg/dl (3.24 mmol/l) had a statistically significant increased risk of an all-cause mortality, including those taking or not taking lipid-modifying medications, compared with the reference of 176-225 mg/dl (4.54-5.83 mmol/l). In stratified analysis, this association was demonstrated in patients with serum albumin >3.0 g/dl (30 g/l), but not with albumin < or =3.0 g/dl. Compared with patients with triglyceride levels of 201-300 mg/dl (2.27-3.39 mmol/l), a statistically significant reduction of all-cause, but not CV, mortality was observed in patients with triglyceride levels of 101-200 mg/dl (1.14-2.26 mmol/l), as well as in the subgroup with serum albumin levels <3.0 g/dl (30 g/l) and triglycerides of < or =100 mg/dl (1.13 mmol/l) and 101-200 mg/dl (1.14-2.26 mmol/l). CONCLUSIONS: While confounding factors and causal pathways have not been clearly identified, aggressive lowering of plasma cholesterol in PD patients is not supported by this study, however, treatment of hypertriglyceridaemia may be warranted with triglyceride levels >200 mg/dl (2.26 mmol/l).     

Expression of two isozymes of acyl-coenzyme A: Cholesterol acyltransferase-1 and -2 in clear cell type renal cell carcinoma

Objectives:   The present study investigated the expression of acyl-coenzyme A: cholesterol acyltransferase-1 (ACAT-1) and -2 in clear cell type renal cell carcinoma (RCC).
Methods:   Clear cell type RCC and corresponding normal kidney tissue samples were obtained from 19 surgical cases (28–85 years of age). Tissue extracts were assayed for ACAT activity and protein expression by immunoblotting with anti-ACAT-1 and anti-ACAT-2 antibodies. Frozen sections were subjected to Oil red O staining for lipids, and were immunostained with ACAT-specific antibodies.
Results:   Acyl-coenzyme A cholesterol acyltransferase activity was 5.7-fold higher ( P  < 0.01) in clear cell carcinoma (23.52 ± 4.90 pmol/mg protein/min) than in normal kidney (4.12 ± 0.36). Consistent with this, immunoblotting and immunohistochemical staining revealed strong expression of ACAT-1 in clear cell type RCC. Densitometric analysis showed that ACAT-1 expression was 2.9-fold higher in clear cell type RCC than in normal kidney. In contrast, ACAT-2 expression was negative in clear cell type RCC and normal kidney. Oil red O staining showed massive deposits of lipid in RCC cells.
Conclusions:   We identified strong expression of ACAT-1 in clear cell type RCC. Upregulation of ACAT-1 leads to high ACAT enzymatic activity, which accelerates the accumulation of cholesterol ester in clear cell type RCC.     

A clinicopatholgoical study on intrahepatic cholesterol gallstones

In order to clarify the pathogenesis and process of the formation of intrahepatic cholesterol gallstones, we examined the clinical features, cholangiograms and pathological findings of eight patients with intrahepatic cholesterol gallstones. When examining the clinical features, one patient was found to have developed intrahepatic cholesterol gallstones 3 years after a complete lithotomy. The cholangiograms of two patients revealed small gallstones in the peripheral bile ducts of the lateral segment of the liver, and these bile ducts showed localized cystic dilatation and were tightly filled with gallstones. Conversely, their other bile ducts which contained no gallstones showed an entirely normal cholangiogram. Pathologically, these two cases showed mild chronic cholangitis, and cholesterol crystals in the peripheral bile ducts. The other six cases showed moderate or severe dilatation of the bile duct and severe chronic proliferative cholangitis. From the above results, we proposed the following theory to explain the pathogenesis and process of the formation of intrahepatic cholesterol stones: The cholesterol crystals in the peripheral intrahepatic bile ducts may be a primitive form of intrahepatic cholesterol gallstones, and the formation of intrahepatic cholesterol gallstones may precede and cause such deformities of the bile ducts as strictures or dilatations.