Insulin-like growth factor I (IGF-I) in a growth-enhanced transgenic (GH-overexpressing) bony fish,the tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>): indication for a higher impact of autocrine/paracrine than of endocrine IGF-I

Several lines of growth hormone (GH)-overexpressing fish have been produced and analysed for growth and fertility parameters. However, only few data are available on the growth-promoting hormone insulin-like growth factor I (IGF-I) that mediates most effects of GH, and these are contradictory. Using quantitative real-time RT-PCR, radioimmunoassay, in situ hybridization, immunohistochemistry, and radiochromatography we investigated IGF-I and IGF binding proteins (IGFBPs) in an adult (17 months old) transgenic (GH-overexpressing) tilapia (Oreochromis niloticus). The transgenics showed an around 1.5-fold increase in length and an approximately 2.3-fold higher weight than the non-transgenics. Using radioimmunoassay, the serum IGF-I levels were lower (6.22 ± 0.75 ng/ml) in transgenic than in wild-type (15.01 ± 1.49 ng/ml) individuals (P = 0.0012). Radioimmunoassayable IGF-I in transgenic liver was 4.2-times higher than in wild-type (16.0 ± 2.21 vs. 3.83 ± 0.71 ng/g, P = 0.0017). No hepatocytes in wild-type but numerous hepatocytes in transgenic liver contained IGF-I-immunoreactivity. RT-PCR revealed a 1.4-times higher IGF-I mRNA expression in the liver of the transgenics (10.51 ± 0.82 vs. 7.3 ± 0.49 pg/μg total RNA, P = 0.0032). In correspondence, in situ hybridization showed more IGF-I mRNA containing hepatocytes in the transgenics. A twofold elevated IGF-I mRNA expression was determined in the skeletal muscle of transgenics (0.33 ± 0.02 vs. 0.16 ± 0.01 pg/μg total RNA, P < 0.0001). Both liver and serum of transgenics showed increased IGF-I binding. The increased IGFBP content in the liver may lead to retention of IGF-I, and/or the release of IGF-I into the circulation may be slower resulting in accumulation of IGF-I in the hepatocytes. Our results indicate that the enhanced growth of the transgenics likely is due to enhanced autocrine/paracrine action of IGF-I in extrahepatic sites, as shown here for skeletal muscle.     

Type 1 Diabetic Serum Interferes with Pancreatic β-cell Ca2+ -Handling

The aim of this study was to clarify the frequency of patients with type 1 diabetes that have serum that increases pancreatic β-cell cytoplasmic free Ca²⁺ concentration, [Ca²⁺]_i, and if such an effect is also present in serum from first-degree relatives. We also studied a possible link between the serum effect and ethnic background as well as presence of autoantibodies. Sera obtained from three different countries were investigated as follows: 82 Swedish Caucasians with newly diagnosed type 1 diabetes, 56 Americans with different duration of type 1 diabetes, 117 American first-degree relatives of type 1 diabetic patients with a mixed ethnic background and 31 Caucasian Finnish children with newly diagnosed type 1 diabetes. Changes in [Ca²⁺]_i , upon depolarization, were measured in β-cells incubated overnight with sera from type 1 diabetic patients, first-degree relatives or healthy controls. Our data show that there is a group constituting approximately 30% of type 1 diabetic patients of different gender, age, ethnic background and duration of the disease, as well as first-degree relatives of type 1 diabetic patients, that have sera that interfere with pancreatic β-cell Ca²⁺-handling. This effect on β-cell [Ca²⁺]_i could not be correlated to the presence of autoantibodies. In a defined subgroup of patients with type 1 diabetes and first-degree relatives a defect Ca²⁺-handling may aggravate development of β-cell destruction.     

Rapid and direct determination of selenium, copper, and zinc in blood plasma by flow injection-inductively coupled plasma-mass spectrometry

A flow injection-inductively coupled plasma-mass spectrometry (FI-ICP-MS) with a simple sample preparation procedure was developed for the determination of selenium, copper, and zinc in blood serum/plasma. A serum/plasma sample was filtered through a 0.45-μm membrane filter and diluted with a mixture of trace elements in a standard solution (9∶1, v/v). Measurement of the reference serum sample confirmed the accuracy of our method for selenium, copper, and zinc concentration. In the case of blood plasma samples obtained from six healthy adult males, the selenium, copper, and zinc concentrations were similar to those of a typical healthy male in Japan. These results suggest that the sample prepartive procedure coupled with FI-ICP-MS can be used for the routine determination of selenium, copper, and zinc in human blood serum/plasma.     

A novel radioimmunoassay of 7-oxo-DHEA and its physiological levels

A novel radioimmunoassay (RIA) of unconjugated 7-oxo-dehydroepiandrosterone (7-oxo-DHEA) in human serum was developed for the first time. This steroid is an intermediate in the biosynthesis of immunomodulatory 7-hydroxylated DHEA metabolites, and has been shown to possess thermogenic properties. The method employs polyclonal rabbit antiserum to (19E)-3beta-hydroxy-7,17,19-trione-19-O-(carboxymethyloxime):BSA conjugate and a homologous radioiodinated derivative with tyrosine methyl ester. The cross reactivity of the antiserum with structurally closest 7-hydroxyepimers of DHEA was lower than 1.7%, with DHEA 0.4%, with all other related steroid less than 0.4%. The method includes ether extraction of serum (0.5 ml), followed by RIA. Its detection limit was 0.06 pmol (18 pg)/tube, the average intra- and inter-assay coefficients of variation were 4.1% and 8.3%, respectively. Mean recovery of serum spiked with 7-oxo-DHEA varied between 78.8% and 112%. Its levels in three serum pools were compared with a low-resolution gas chromatography-mass spectrometry method with satisfactory results. The method has been used for determination of 7-oxo-DHEA in serum samples of 215 subjects (91 males and 124 females) without overt endocrine disorders, aged 5-71 years. The over-all mean+/-S.D. was 0.280+/-0.227, the median 0.239 nmol/l. No significant sex differences were recorded. The only group which differed significantly from all other ones were males below 10 years, significantly lower values than in other age groups were found also in the first two age groups of females.     

肠道菌群失调与脂肪肝患者血脂和丙氨酸氨基转移酶的变化研究

目的探讨脂肪肝患者体内甘油三酯(TG)、血清总胆固醇(CHOL)、高密度脂蛋白(HDL)、低密度脂蛋白(LDL)、丙氨酸氨基转移酶(ALT)与肠道菌群失调的关系。方法单一他汀类降脂药物治疗高脂血症,测定肠道菌群失调与肠道菌群正常的脂肪肝患者血清TG、CHOL、HDL、LDL、ALT。结果无菌群失调组,ALT升高者占11%,治疗缓解率为87%,而菌群失调组ALT升高占37%,治疗缓解率为67%,两者差异有显著性。结论正常肠道的菌群对血脂代谢以及高脂血症的治疗起着重要作用。     

Differential radioactive quantification of protein abundance ratios between benign and malignant prostate tissues: cancer association of annexin A3

A differential quantitative protein expression study, comparing matched prostate cancerous and benign tissues from 31 patients, revealed proteins newly associated with prostate cancer. Average effects for 17 proteins whose abundance was significantly different (p<0.01) across patients ranged from 1.5- to 6.1-fold, and included a number of known cancer markers. The most differentially abundant proteins between cancer and benign samples were isopeptidase T, serum amyloid P (SAP), annexin A3 (ANXA3) and mitochondrial enoyl coenzyme-A hydratase. SAP is restricted to stroma in healthy tissue, and the lower abundance in tumours may be explained by the reduced stromal content. ANXA3 is present in healthy epithelial cells, exhibits strong staining in precancerous prostatic intraepithelial neoplasia, and is relatively less abundant in individual tumour cells of increasing Gleason pattern (GP), despite exhibiting higher overall tissue abundance in tumours. ANXA3 staining was predominantly cytoplasmic, yet nuclear localization was also observed. Strongly staining single cells, possibly phagocytes, were interspersed in highly dedifferentiated GP5 tumour areas among tumour cells without measurable ANXA3. Local recurrent androgen ablation therapy-resistant tumours exhibit heterogenous low levels of ANXA3 staining. Results are discussed focussing on the potential implications for tumour tissues.     

A neuronal cell-based botulinum neurotoxin assay for highly sensitive and specific detection of neutralizing serum antibodies

Clostridium botulinum neurotoxin (BoNT) serotypes A and B are widely used as pharmaceuticals to treat various neurological disorders and in cosmetic applications. The major adverse effect of these treatments has been resistance to treatment after multiple injections. Currently, patients receiving BoNT therapies and patients enrolled in clinical trials for new applications and/or new formulations of BoNTs are not routinely monitored for the formation of neutralizing antibodies, since no assay other than the mouse protection procedure is commercially available that reliably tests for the presence of such antibodies. This report presents a highly sensitive and specific neuronal cell-based assay that provides sensitive and specific detection of neutralizing antibodies to BoNT/A.     

快速序贯器官衰竭评分联合血清同型半胱氨酸、三酰甘油对急性胰腺炎短期预后的预测价值研究

摘要 目的：探讨快速序贯器官衰竭评分（qSOFA）联合血清同型半胱氨酸（Hcy）、三酰甘油（TG）对急性胰腺炎（AP）短期预后的预测价值。方法：选取2019年9月至2021年9月苏州大学附属第一医院消化科收治的210例AP患者为研究对象,根据亚特兰大AP分类指南分为轻症AP（MAP组）125例,中重症AP（MSAP组）45例和重症AP（SAP组）40例。对比三组的qSOFA评分和Hcy、TG水平。根据入院后28d预后结局,将所有患者分为存活组192例、死亡组18例。单因素及多因素Logistic回归分析AP患者预后的影响因素。采用受试者工作特征（ROC）曲线分析qSOFA评分、Hcy和TG对AP患者短期预后的预测价值。结果：SAP组和MSAP组患者的qSOFA评分、Hcy、TG水平均高于MAP组患者,且SAP组高于MSAP组（P＜0.05）;单因素、多因素Logistic回归最终分析结果显示,qSOFA评分较高、TG、Hcy水平升高是AP患者死亡的危险因素（P＜0.05）;qSOFA评分联合Hcy、TG预测AP患者短期预后的曲线下面积（AUC）为0.982,明显高于三指标单独检测的0.715、0.780、0.782。结论：qSOFA评分、TG、Hcy水平均是AP患者短期预后的影响因素,并且联合检测qSOFA评分和Hcy、TG水平对AP患者的短期预后具有较高的预测价值。     

帕利哌酮治疗伴有精神病性症状的抑郁发作患者的疗效分析及对神经功能及血清BDNF的影响

摘要 目的：探讨帕利哌酮治疗伴有精神病性症状的抑郁发作患者的疗效分析及对神经功能及血清脑源性神经营养因子（BDNF）的影响。方法：选取本院2021年1月到2021年10月收治的100例伴有精神病性症状的抑郁发作患者作为研究对象,随机将其分为观察组（n=50）和对照组（n=50）。对照组采用常规药物为伴有精神病性症状的抑郁发作患者进行治疗,观察组在对照组的基础上采用帕利哌酮为伴有精神病性症状的抑郁发作患者进行治疗,对比两组患者治疗前、治疗后2周、治疗后6周的汉密尔顿焦虑量表（HAMD）评分、汉密尔顿抑郁量表（HAMA）评分、精神经功能缺损程度、血清BDNE、神经病评定量表（BPRS）评分、社会功能缺陷筛选量表（SDSS）评分、日常生活能力量表（ADL）评分以及不良反应发生率。结果：治疗前两组患者的HAMA评分和HAMD评分对比无明显差异（P>0.05）,治疗后2周、6周评分均降低,且观察组较对照组低（P＜0.05）;治疗前两组患者的NIHSS评分和血清BDNE水平对比无明显差异（P>0.05）,治疗后2周、6周两组患者的NIHSS评分均低,且相较于观察组,对照组较高（P＜0.05）,但血清BDNE水平均升高,且观察组较对照组高（P＜0.05）;治疗前两组患者的BPRS、SDSS、ADL评分对比明显差异（P＞0.05）。治疗后2周、6周两组患者的BPRS、SDSS皆降低,并且观察组低于对照组（P＜0.05）,但两组患者的ADL评分均升高并且观察组高于对照组（P＜0.05）;观察组患者的不良反应总发生率与对照组比较无差异（P>0.05）。结论：将帕利哌酮应用于伴有精神病性症状的抑郁发作患者当中,可改善患者的焦虑、抑郁以及神经功能缺损情况,提高血清BDNE水平,并降低神经病性和社会功能缺陷情况,提高患者日常生活能力,值得临床借鉴。     

磁共振成像联合血清前列腺特异抗原、上皮钙黏蛋白、早期前列腺癌抗原-2诊断前列腺癌的临床价值探讨

摘要 目的：探究磁共振成像（MRI）联合血清前列腺特异抗原（PSA）、上皮钙黏蛋白（sE-cadherin）、早期前列腺癌抗原-2（EPCA-2）诊断前列腺癌的临床价值。方法：选取潍坊市人民医院2020年1月-2021年7月期间经病理证实的50例前列腺癌患者（前列腺组）以及50例前列腺增生患者（前列腺增生组）展开回顾性研究。100例研究对象均完善了MRI检查并测定血清PSA、EPCA-2、sE-cadherin水平,分析两组患者的MRI影像学特征,比较两组患者的PSA、EPCA-2、sE-cadherin水平以及各项检查方法的诊断准确性差异。结果：前列腺癌的MRI影像学特征为病灶主要位于外周带,外周带T2W呈低信号,病变侵及包膜、膀胱及周围组织具有T1加权消失或者不对称,具有信号异常、肌肉增厚的表现,盆腔淋巴结转移具有淋巴结部分融合或增大表现;前列腺增生的MRI影像学特征为边界清晰、包膜完整并且中央带增生、不均匀信号结节;前列腺癌、前列腺增生均存在不同程度的前列腺体积增大。相比于前列腺增生组,前列腺癌组患者的PSA、sE-cadherin、EPCA-2水平明显更高（P<0.05）。MRI、PSA、sE-cadherin、EPCA-2四项联合鉴别前列腺癌、前列腺增生的诊断符合率为96.00%,明显高于四项单独诊断的88.00%、79%、81%、82%（P<0.05）。结论：MRI联合PSA、sE-cadherin、EPCA-2鉴别诊断前列腺癌的准确性较高,具有作为临床前列腺癌早期诊断指导方案的潜力。