升清胶囊对肝细胞氧化损伤模型中胆固醇代谢相关酶表达的影响

目的:探讨升清胶囊对人肝细胞氧化损伤模型中胆固醇7α-羟化酶(CYP7A1)和固醇携带蛋白2(SCP2)m RNA及蛋白表达的影响。方法:选用人L-02肝细胞,分为正常组、模型组、空白血清组及升清胶囊组。采用H2O2诱导建立氧化损伤模型,空白血清组和升清胶囊组每孔分别加入2 m L含10%血清或含药血清的RPMI-1640培养液,培养24 h后采用real-time PCR法和western-blot法分别检测各组肝细胞CYP7A1和SCP2 m RNA及蛋白表达。结果:与正常组比较,模型组CYP7A1 m RNA及蛋白表达降低,SCP2 m RNA及蛋白表达升高(P<0.01);与模型组比较,空白血清和升清胶囊组CYP7A1m RNA及蛋白表达升高,SCP2 m RNA及蛋白表达降低(P<0.01);与空白血清组比较,升清胶囊组CYP7A1 m RNA及蛋白表达增强,SCP2 m RNA及蛋白表达降低(P<0.05)。结论:升清胶囊可以通过下调SCP2 m RNA及蛋白表达来减少肝细胞分泌胆固醇入胆汁,同时上调CYP7A1 m RNA及蛋白表达来加速胆汁酸生成,从而防止胆固醇结晶析出,达到降低胆结石形成的作用。     

胆汁促成核糖蛋白的分离、提纯及理化性质研究

胆汁中促、抗成核因子的发现是近年来胆固醇性结石成因研究中的一大进展。我们应用伴刀豆素Ａ凝集素固相化亲和层析结合ＳｅｐｈａｄｅｘＧ－２００分于筛色谱技术，于１９９１年底从人体胆汁中分离和提纯得到一种糖蛋白，分子量为７０ＫＤ，具有明显的促成核活性，其中性已糖与蛋白质含量之比为１５．８８％，一分子量的糖蛋白约含６４０个氨基酸，其Ｎ－末端氨基酸顺序为Ｎ－Ｓｅｒ－Ｈｉｓ－Ｇｌｙ－Ｍｅｔ－Ａｒｇ－Ｇｌｎ－Ｔｙｒ－Ｔｙｒ－Ｍｅｔ－Ｌｙｓ。本文同时讨论了这一糖蛋白对胆石成因的病理学和生物学意义。     

胆汁成核效应蛋白的分离纯化与成核活性比较

目的 纯化并比较各种成核效应蛋白的活性,筛选最具病理学意义的促成核因子。方法 通过ConA-Sepharose 4B亲和层析和Sephadex G-200、Sephadex G-100分子筛凝胶过滤以及超速离心法从胆固醇性结石病人胆汁中分离提纯70 kDa、200 kDa促成核蛋白以及33.5 kDa泡蛋白,在模拟胆汁体系中观察成核因子促进胆固醇结晶生长、析出过程,比较其成核活性。结果 四种候选的成核效应蛋白中,33.5 kDa泡蛋白是活性最强的促成核因子,成核活性强弱依次为33.5 kDa泡蛋白>200 kDa糖蛋白≈α1-酸性糖蛋白>70 kDa糖蛋白,当浓度为100／μg／ml时,33.5 kDa泡蛋白成核活性指数Ig、Ic、It分别为1.52、1.63、0.57,并随着蛋白浓度的增加,促成核活性亦随之增强(F=678.18,P<0.05),且呈剂量依赖效应。结论 胆汁中存在多种促成核作用的糖蛋白,其中33.5 kDa泡蛋白可能与胆石形成关系最为密切。     

33.5KDa胆汁泡蛋白促成核活性及其对泡形态学影响的研究

目的探讨３３．５ｋＤａ泡蛋白在胆固醇成核过程中的作用。方法利用超速离心技术结合ＰＡＧＥ电泳及区带定位法制备３３．５ｋＤａ泡蛋白；并将其加入Ｓｍａｌ模拟胆汁及综合模拟胆汁作为试验组，相应地以人体白蛋白为对照组，用偏光显微镜观察胆固醇成核时间，在透射电镜下观察模拟胆汁泡的大小、形态及数量。结果Ｓｍａｌ试验组成核时间为３．８３±０．３１天，对照组为１２．３３±０．５６天（Ｐ＜０．００１）；综合试验组平均成核时间为３．１７±０．１７天，而对照组长达１０．３３±０．２１天（Ｐ＜０．００１）。随着时间的推移，Ｓｍａｌ试验组及综合试验组泡逐渐增大、聚集、融合，最终形成巨大复层泡。两组均较相应的对照组变化显著（Ｐ＜０．０５）。结论３３．５ｋＤａ泡蛋白具有很强的促进泡形成、聚集、融合及胆固醇单水结晶析出的作用（成核活性约为０．３１０）。     

胆固醇结石病人胆汁免疫球蛋白含量增高及其对胆固醇-磷脂泡的影响

目的 :研究胆固醇结石病人胆囊胆汁、胆总管胆汁、血清3种免疫球蛋白(immunoglobulin,Ig)含量及其对胆固醇-磷脂泡变化的影响。方法:通过Con A亲和层析与Sephadex G-200分子筛凝胶两次过滤,从胆囊胆固醇结石病人及胆囊胆色素结石病人、无结石对照病人、非结石急性胆囊炎病人以及白胆汁病人胆囊胆汁、胆总管胆汁和血清中分离提纯Ig A、Ig G和Ig M;应用单向免疫扩散法检测3种Ig含量;应用电子显微镜观察Ig在模拟胆汁体系对胆固醇-磷脂泡的影响。结果:胆固醇结石组胆囊胆汁含有较高浓度的Ig G和Ig M,胆总管胆汁的3种Ig含量增高。胆色素结石组胆总管胆汁Ig A、Ig M含量与对照组无差异。胆固醇结石组和胆色素结石组除血清Ig A含量低于对照组外,其余Ig含量与对照组无差异。胆固醇结石组胆囊胆汁Ig G和Ig M分别作用于模拟胆汁7 d,明显促使单层泡直径增大,且增加多层泡数量;而Ig A仅轻微增加单层泡直径,不影响多层泡数量。结论:本实验结果显示,胆固醇结石组胆汁中Ig增高,可能通过影响胆固醇-磷脂泡变化,促进胆固醇结石形成。     

胆汁中蛋白成分对胆固醇结石形成的影响

胆囊胆固醇结石的形成常与胆汁成分改变有关，但其确切机制及其始动因素尚未完全明了，该文就近年胆汁中蛋白质成分改变对胆石形成的影响及其可能机制方面研究成果作一综述。     

熊去氧胆酸对胆固醇结石患者胆囊胆汁脂类浓度成核时间及胆囊排空能力的影响

作者测定了38例胆囊胆固醇结石患者服用小量熊去氧胆酸(400mg/d)后的胆囊胆汁脂类浓度、成核时间及胆囊排空能力的变化。结果发现：小剂量熊去氧胆酸可降低胆囊胆汁胆固醇饱和指数，延长成核时间。但对胆囊排空能力无影响。认为熊去氧胆酸可通过降低胆固醇饱和指数．延长胆固醇晶体成核过程而对胆囊结石起预防作用。     

升清胶囊对阻塞性黄疸患者微创术后细胞免疫功能的改善作用

目的 研究升清胶囊对阻塞性黄疸患者微创术后细胞免疫功能的改善作用.方法 将符合纳入标准的51例患者,采用简单随机方法分为研究组(升清胶囊组)、对照组(胆维他组)和空白组(常规治疗组),每组17例.术后空白组常规处理,不服用利胆药物;对照组除常规处理外,于术后第1天起口服胆维他;研究组除常规处理外,于术后第1天开始口服中药升清胶囊.术后第1、7天分别检测患者免疫指标CD3、CD4、sIL-2R、IL-10、TNF及TB.采用单因素方差分析的统计学方法对实验数据做统计处理.结果 研究组、对照组与空白组术后第1天CD3、CD4、sIL-2R、TNF、IL-10值比较均无统计学差异(P>0.05);术后第7天比较均有统计学差异(P<0.05).术后第7天CD.CD4、sIL-2R值与术后第1天比较,研究组比对照组均增高(P<0.05);时照组比空白组均增高(P<0.05);研究组比空白组均增高(P<0.05).术后第7天TNF值与术后第1天比较,研究组较对照组无显著性降低(P>0.05);对照组比空白组降低(P<0.05);研究组比空白组降低(P<0.05).术后第7天IL-10值与术后第1天比较,研究组比对照组降低(P<0.05);对照组比空白组无显著性降低(P>0.05);研究组比空白组降低(P<0.05).术后第1、7天各组间TB值比较均无统计学差异(P>0.05).结论 升清胶囊能更有效地加快阻塞性黄疸患者微创术后免疫功能的恢复,较不服用利胆药物及服用胆维他者有明显优势.     

胃大部切除术对骨密度和生化指标的影响

鉴于有人提出胃切除术后会伴有骨代谢的改变，我们研究的目的就是评定经胃大部切除术（毕Ⅱ式）治疗的溃疡病患者的骨密度和骨代谢生化指标的变化。我们选择了３０名３０～６５岁的患者并与年龄、性别相配的以常规药物治疗的消化道溃疡组患者作对照。我们发现骨密度在患者组显著低于对照组，降钙素水平患者组亦显著低于对照组。但血清钙、磷、镁、骨钙素；尿钙、磷、镁在两组间没有显著差异。胃切除术引起骨质减少的机制仍需全面阐明。我们研究的结果表明胃大部切除术后椎骨密度有显著降低。生化指标没有变化。但降钙素明显降低。骨钙素水平有下降趋势，提示成骨细胞活动性减弱。降钙素水平的下降提示骨吸收的增加，进一步提示高骨更新率可能是胃大部切除术后人群骨质减少的一个原因     

大豆异黄酮对大鼠骨密度及骨代谢生化指标的影响

目的 研究植物雌激素-大豆异黄酮对大鼠骨密度及骨代谢生化指标的影响。方法 将断乳Wister大鼠按体重随机分为4组，分别给予基础饲料和不同剂量的大豆异黄酮。每周称体重，调整给食量。12周后处死大鼠，取脏器称重，计算脏体比值；剥离股骨，测骨矿物质含量(BMC)、骨密度(BMD)和骨钙、骨磷的含量；对血清中骨形成生化指标碱性磷酸酶、骨钙素和骨吸收生化指标抗酒石酸酸性磷酸酶进行检测，同时测定雌激素-雌二醇(E2)的含量。结果 具有弱雌激素样作用的大豆异黄酮对实验大鼠的子宫、卵巢无刺激作用。与对照组相比，给予大豆异黄酮能提高BMC、BMD及骨钙含量，并随剂量的增加而增大。大豆异黄酮可影响骨代谢，高剂量的大豆异黄酮(41．6mg／kg)同时抑制骨形成和骨吸收，使骨转化率降低，但对骨吸收的作用大于骨形成。给予大豆异黄酮组血清雌激素水平大于对照组。结论 大豆异黄酮通过调整骨代谢生化指标的活性，提高大鼠的骨钙含量和骨密度，可预防骨质疏松的发生。     

Lower-zone emphysema in young patients without α1-antitrypsin deficiency

Martelli, N. A., Goldman, E., and Roncoroni, A. J. (1974).Thorax, 29, 237-244. Lowerzone emphysema in young patients without α₁-antitrypsin deficiency. Three young patients with radiographic pulmonary emphysema predominantly in the lower zones are reported. The clinical and physiological features were those observed in severe pulmonary emphysema. Predominance of the main lesions in the lower zones was confirmed in two cases by selective pulmonary angiography. One of the patients died and extensive panlobular emphysema was found at necropsy. Although the similarities between our patients and those with emphysema and α₁-antitrypsin deficiency were remarkable, the latter condition was ruled out.     

Clinical features and prognosis of life time non-smokers with severe α1-antitrypsin deficiency

BACKGROUND—The hereditary disorderα₁-antitrypsin deficiency is characterised by developmentof severe emphysema at an early age with smoking being the mostsignificant additional risk factor. The purpose of the present paperwas to analyse potential risk factors other than smoking for emphysemaand to estimate the prognosis of life time non-smokers.
METHODS—Patients were identified through the filesof the Danish α₁-antitrypsin deficiency register whichcontains information on more than 700 persons with the condition. Manyof the patients, the non-index cases, were identified from family studies.
RESULTS—There were 75 life time non-smokers withPiZ (27 index cases and 48 non-index cases) aged 20 years or more atentry. Twenty one subjects died during the follow up period. TheStandardised Mortality Ratio (SMR) was 3.0 (95% confidence intervals(CI) 1.9to 4.6). There was no significant difference in SMR betweenmales and females. The SMR was 8.8 (95% CI 5.0 to 14) for the index cases and 0.96 (95% CI 0.3 to 2.3) for the non-index cases based onfive deaths. The overall mean % predicted forced expiratory volume inone second (FEV₁) at entry was 83% with a significant difference between index cases (54%) and non-index cases (100%) (p<0.001). The difference in the ratio of FEV₁ to forcedvital capacity (FVC) was also highly significant with values of 0.57and 0.79 for index and non-index cases, respectively (p<0.001). In thenon-index group only three had an FEV₁% predicted of less than 70%.
CONCLUSIONS—Occupational exposure to airwayirritants did not have any significant influence on the development ofemphysema. Only a few life time non-smokers develop severe emphysema;most never develop pulmonary symptoms and thus remain undetected unlessfamily members of index cases are screened.

     

Bacteriological investigation of bile in patients with cholelithiasis

The microflora in bile from the gallbladder and common bile duct was investigated in 303 patients who underwent surgery for cholelithiasis. The purpose of this study was to identify current bacteria and bacterial casts in the biliary tract and also to analyze the relationship between bactericholia at the time of operation and postoperative infection. Bile cultures were positive in 38% of all patients, although a higher incidence of positive bile cultures occurred in patients over 70 years of age (77%), those with common duct stones (83%), those with pigment stones (65%), and those who underwent gastrectomy (71%). The predominant organisms were Escherichia coli (22%), Klebsiella (18%), and Enterococcus (15%). Obligate anaerobes were less frequently seen (4%), being found only in patients with pigment stones and always mixed with aerobes. Four patients developed postoperative infections (1.3%) which were all caused by biliary bacteria. The following two factors may contribute to this low incidence of postoperative infections: our policy of operating electively whenever possible, and the prophylactic use of antibiotics to which the organisms cultured from bile are sensitive.     

Evaluation of autoantibodies against vimentin and α-enolase in rheumatoid arthritis patients

IntroductionRheumatoid arthritis (RA) is categorized as an autoimmune disease with a frequency of 0.2–1% worldwide. It is reported that various autoantibodies are produced in the RA population, particularly against citrullinated peptides. Among various candidate markers for RA diagnosis, the citrullinated proteins have the highest specificity and sensitivity for both diagnosis and prognosis of RA. Anti-mutated citrullinated vimentin and α-enolase constitute a new class of autoantibodies for early detection of RA.Material and methods45 serum samples and 19 synovial fluid (SF) specimens collected from RA patients were considered for American College of Rheumatology criteria and 20 serum samples and 10 SF specimens were provided from healthy subjects as a control group. To assess the quantity of anti-citrullinated protein antibodies (ACPA), anti-mutated citrullinated vimentin (MCV) and anti-α-enolase in the serum and SF of RA patients were determined by the enzyme-linked immunosorbent assay (ELISA) method. For the evaluation of disease activity and joint destruction, we used the Disease Activity Score of 28 joints based on erythrocyte sedimentation rate (ESR) Disease Activity Score 28 (DAS28). Furthermore, to measure the molecular weight of vimentin and α-enolase, electrophoresis on 10% SDS-PAGE was performed as described before.ResultsThe anti-α-enolase level among serum samples from RA patients was significantly higher than in healthy subjects (4.49 ±0.20 ng/ml vs. 0.76 ±0.12 ng/ml) (p < 0.001). There was a direct relation between α-enolase quantity and (rheumatoid factor) RF and C-reactive protein (CRP) levels. The mean ESR value in positive and negative ACPA patients was 38.2 ±22.6 mm/h and 9.2 ±5.8 mm/h respectively (p < 0.0001). The mean DAS28-ESR was 3.3. The level of anti-MCV in the serum of RA patients (244.6 ±53.3 U/ml) was higher than in serum of the healthy group (148.73 ±71.8) (p < 0.0001). The level of anti-MCV in the SF of patients was 687.5 ±148.4 U/ml.ConclusionsIn conclusion, both autoantibodies against MCV and α-enolase are two important markers that increase in serum and SF of RA patients and are specific for diagnosis of RA disease.     

Biophysical properties of bile and urine in patients with cholelithiasis

The electric conductivity of the hepatic bile received during and 24 hours after surgery from the drainage introduced into the common hepatic duct was studied in 94 patients with cholelithiasis. It is demonstrated that conductivity increases in cholestasis, inflammatory process in the biliary tract and depends on the hepatic morphofunctional status. The specific electric conductivity of intraoperative bile that is more than 192 S/m is considered to be criteria for the diagnosis of cholangitis. The electric conductivity of urine before and in first 4 days after surgery decreases in inflammation of the biliary tract and depends on the degree of concomitant renal dysfunction and electrolyte disturbances. There was a decrease in the conductivity of bile and urine in all studied groups. Detection of biophysical parameters of biological fluids may objectively control the postoperative period and correct treatment policy.     

Conversion of α-Cells to β-Cells in the Postpartum Mouse Pancreas Involves Lgr5 Progeny

In contrast to the skin and the gut, where somatic stem cells and their niche are well characterized, a definitive pancreatic multipotent cell population in the adult pancreas has yet to be revealed. Of particular interest is whether such cells may be endogenous in patients with diabetes, and if so, can they be used for therapeutic purposes? In the current study, we used two separate reporter lines to target Cre-recombinase expression to the Lgr5- or glucagon-expressing cells in the pancreas. We provide evidence for the existence of a population of cells within and in the proximity of the ducts that transiently express the stem-cell marker Lgr5 during late gestational stages. Careful timing of tamoxifen treatment in Lgr5^{EGFP-IRES-CreERT2};R26^Tomato mice allowed us to show that these Lgr5-expressing progenitor cells can differentiate into α-cells during pregnancy. Furthermore, we report on a spontaneous lineage conversion of α- to β-cells specifically after parturition. The contribution of Lgr5 progeny to the β-cell compartment through an α-cell intermediate phase early after pregnancy appears to be part of a novel mechanism that would counterbalance against excessive β-cell mass reduction during β-cell involution.     

Deletion of the Brain-Specific α and δ Isoforms of Adapter Protein SH2B1 Protects Mice From Obesity

Mice lacking SH2B1 and humans with variants of SH2B1 display severe obesity and insulin resistance. SH2B1 is an adapter protein that is recruited to the receptors of multiple hormones and neurotrophic factors. Of the four known alternatively spliced SH2B1 isoforms, SH2B1β and SH2B1γ exhibit ubiquitous expression, whereas SH2B1α and SH2B1δ are essentially restricted to the brain. To understand the roles for SH2B1α and SH2B1δ in energy balance and glucose metabolism, we generated mice lacking these brain-specific isoforms (αδ knockout [αδKO] mice). αδKO mice exhibit decreased food intake, protection from weight gain on standard and high-fat diets, and an adiposity-dependent improvement in glucose homeostasis. SH2B1 has been suggested to impact energy balance via the modulation of leptin action. However, αδKO mice exhibit leptin sensitivity that is similar to that of wild-type mice by multiple measures. Thus, decreasing the abundance of SH2B1α and/or SH2B1δ relative to the other SH2B1 isoforms likely shifts energy balance toward a lean phenotype via a primarily leptin-independent mechanism. Our findings suggest that the different alternatively spliced isoforms of SH2B1 perform different functions in vivo.     

Regulation of the effects of TGF-β1 by activation of latent TGF-β1 and differential expression of TGF-β receptors (TβR-I and TβR-II) in idiopathic pulmonary fibrosis

BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is characterised by subpleural fibrosis that progresses to involve all areas of the lung. The expression of transforming growth factor-beta1 (TGF-beta 1), a potent regulator of connective tissue synthesis, is increased in lung sections of patients with IPF. TGF-beta 1 is generally released in a biologically latent form (L-TGF-beta 1). Before being biologically active, TGF-beta must be converted to its active form and interact with both TGF-beta receptors type I and II (T beta R-I and T beta R-II). TGF-beta latency binding protein 1 (LTBP-1), which facilitates the release and activation of L-TGF-beta 1, is also important in the biology of TGF-beta 1. METHODS: Open lung biopsy samples from patients with IPF and normal controls were examined to localise T beta R-I, T beta R-II, and LTBP-1. Alveolar macrophages (AM) and bronchoalveolar lavage (BAL) fluid were examined using the CCL-64 bioassay to determine if TGF-beta is present in its active form in the lungs of patients with IPF. RESULTS: Immunoreactive L-TGF-beta 1 was present in all lung cells of patients with IPF except for fibroblasts in the subepithelial regions of honeycomb cysts. LTBP-1 was detected primarily in AM and epithelial cells lining honeycomb cysts in areas of advanced IPF. In normal lungs LTBP-1 immunoreactivity was observed in a few AM. AM from the upper and lower lobes of patients with IPF secreted 1.6 (0.6) fmol and 4.1 (1.9) fmol active TGF-beta, respectively, while AM from the lower lobes of control patients secreted no active TGF-beta (p< or =0.01 for TGF-beta in the conditioned media from AM obtained from the lower lobes of IPF patients v normal controls). The difference in percentage active TGF-beta secreted by AM from the lower lobes of patients with IPF and the lower lobes of control patients was significant (p< or =0.01), but the difference between the total TGF-beta secreted from these lobes was not significant. The difference in active TGF-beta in conditioned media of AM from the upper and lower lobes of patients with IPF was also not statistically significant. BAL fluid from the upper and lower lobes of patients with IPF contained 0.7 (0.2) fmol and 2.9 (1.2) fmol active TGF-beta, respectively (p< or =0.03). The percentage of active TGF-beta in the upper and lower lobes was 17.6 (1.0)% and 78.4 (1.6)%, respectively (p< or =0.03). In contrast, BAL fluid from control patients contained small amounts of L-TGF-beta. Using immunostaining, both T beta R-I and T beta R-II were present on all cells of normal lungs but T beta R-I was markedly reduced in most cells in areas of honeycomb cysts except for interstitial myofibroblasts in lungs of patients with IPF. TGF-beta 1 inhibits epithelial cell proliferation and a lack of T beta R-I expression by epithelial cells lining honeycomb cysts would facilitate repair of the alveoli by epithelial cell proliferation. However, the presence of both T beta Rs on fibroblasts is likely to result in a response to TGF-beta 1 for synthesis of connective tissue proteins. Our findings show that biologically active TGF-beta 1 is only present in the lungs of patients with IPF. In addition, the effects of TGF-beta 1 on cells may be further regulated by the expression of T beta Rs. CONCLUSION: Activation of L-TGF-beta 1 and the differential expression of T beta Rs may be important in the pathogenesis of remodelling and fibrosis in IPF.