应用蛋白指纹图谱技术筛选原发性胆汁性肝硬化患者血清特异性标志物

目的: 探讨蛋白指纹图谱技术筛选原发性胆汁性肝硬化(PBC)患者血清中可用于诊断的特异性标志物.方法: 采用弱阳离子纳米磁性微球捕获血清中的蛋白,ProteinChip PBSII-C型蛋白质芯片阅读仪检测绘制成蛋白指纹图谱.所有蛋白指纹图谱采用Biomarker Wizard 3.1分析之后用Biomarker Pattems Sofiware 5.0识别最终可能用于PBC诊断的蛋白标志物并优化组合建立诊断模型.结果: 在PBC患者和对照组之间找到69个差异蛋白峰(P<0.05).其中质荷比(m/z)为3445,4260,8133和16 290的蛋白峰建立PBC诊断模型.该诊断模型能很好地把PBC患者从其他肝脏疾病患者和正常人群中区分出来,其敏感性为93.3%,特异性为95.1%.经双盲实验验证,该模型对PBC诊断的敏感性为92.9%,特异性为82.4%. 结论: 采用纳米磁性微球与蛋白质芯片阅读仪联用的蛋白指纹图谱技术可以检测PBC患者血清中的特异性蛋白标志物,并建立敏感性和特异性均较高的PBC诊断模型.     

SELDI-TOF-MS技术在肝癌诊断和介入治疗评价中的价值

目的:建立肝癌血清学诊断模型,探讨评估SELDI-TOF-MS技术在肝癌诊断和介入治疗评价中的价值.方法:用弱阳离子交换芯片(CM10芯片)和表面增强激光解吸电离飞行时间质谱仪(surface-enhanced laser desorption ionization time-of-flight mass spectrometry,SELDI-TOF-MS)技术,测定60例肝癌患者和60例正常对照者的血清蛋白质指纹图谱,应用BiomarkerWizard统计软件比较肝癌组和正常对照组血清蛋白质表达的差异性,采用Biomarker Pattern软件分析数据建立肝癌诊断模型,比较介入治疗前后血清蛋白质指纹图谱的差异性.结果:在质荷比(M/Z)为2000-10000范围内,和正常血清比较,肝癌的差异峰有3个(M/Z为4182Da、5710Da、6992Da;P<0.01),4182Da和5710Da下调,6992Da上调.用这3个差异蛋白峰建立肝癌诊断模型,诊断肝癌的灵敏度为93.3%(28/30),特异度为90.0%(27/30),正确率为91.7%(55/60),约登指数为0.833.差异蛋白峰(M/Z4182Da)在介入术后1mo明显上调(P<0.05).结论:应用SELDI-TOF-MS技术进行肝癌血清蛋白质指纹图谱分析,建立肝癌诊断树模型,对肝癌的诊断有一定的价值;筛选出的差异蛋白峰对肝癌的介入治疗评估有一定的应用价值.     

应用SELDI-TOF-MS技术建立肝癌筛选血清蛋白质指纹图谱模型

目的:建立肝癌筛选血清蛋白质指纹图谱模型．方法:用表面加强激光解析电离飞行时间质谱技术(SELDI-TOF-MS)及WCX2蛋白芯片获得新发肝癌、肝硬化患者和正常人血清的蛋白质指纹图谱,用计算机软件进行比较分析,建立肝癌的筛选模型．结果:肝癌患者与健康对照组血清蛋白质指纹图谱之间有5个标志蛋白(4477,8943,5181, 8617,13 761 Da)在肝癌患者血清中高表达,肝癌患者与肝硬化患者血清蛋白质指纹图谱之间2个标志蛋白(4477,13 761 Da)在肝癌患者血清中高表达,1个标志蛋白(4097 Da)在肝癌患者血清中低表达．SELDI-TOF-MS技术的特异性(60／60,100％);敏感度(18／20,90％)．分析系统筛选出4477,8943,13 761,4097 Da标志蛋白建立的肝癌诊断模型．结论:建立的血清蛋白质指纹图谱模型能够区分肝癌与非肝癌患者,SELDI-TOF-MS在肝癌的诊断及肿瘤特异性蛋白质生物标志分子的筛选等方面具有一定价值．     

应用SELDI-TOF-MS技术建立SLE患者血清蛋白质指纹图谱筛选模型

目的筛选系统性红斑狼疮（SLE）血清蛋白质指纹图谱模型,并探讨其意义。方法用表面加强激光解析电离飞行时间质谱（SEIDI-TOF-MS）技术获得SLE、类风湿关节炎患者（RA组）、健康人群（对照组）血清中的蛋白质谱,用计算机软件比较分析,建立SLE血清蛋白质指纹图谱筛选模型,并验证该模型诊断SLE的灵敏度和特异性。结果SLE组与非SLE组共有11种蛋白质表达有显著性差异;以其中4个蛋白质生物标志物（5 739 Da、10 832 Da、4 650 Da、15 797 Da）组建的筛选模型,经双盲法验证其灵敏度为84.4%,特异性为90.3%。结论应用SEIDI-TOF-MS技术建立的血清蛋白质指纹图谱模型在SLE的早期诊断中具有一定价值。     

早期肝纤维化外周血清标志物筛选的初步研究

目的探讨早期病毒性乙型肝炎（乙肝）肝纤维化的潜在血清标志物并建立相应的诊断模型。方法采用表面增强激光解吸/离子化时间飞行质谱（SELDI-TOF-MS）技术检测20个健康人和31例早期乙肝肝纤维化患者的血清蛋白质谱,用biomarker Wizard软件筛选出早期肝纤维化的差异表达蛋白质谱峰,用Biomarker Pattern软件对差异蛋白峰值进行线性分析并建立诊断模型。结果在分子量为2000～20000Da的区间内检测出97个峰值,15个为差异蛋白峰,其中早期肝纤维化患者有3个上调,12个下调。用质荷比为M4799和M6125的差异蛋白联合建立的诊断模型最优,其特异度为80%（16/20）,灵敏度为87%（27/31）。结论 SELDI-TOF-MS在筛选早期肝纤维化血清学标志物方面具有较高的灵敏度和特异性,作为肝纤维化早期筛选的方法尚需大宗病例证实及临床后续验证。     

应用血清蛋白质指纹图谱筛选类风湿关节炎的血清标志物

目的 用表面增强激光解吸电离飞行时间质谱（SELDI—TOF—MS）和蛋白质芯片技术检测类风湿关节炎（RA）患者血清蛋白质指纹图谱，探讨基于人工神经网络的蛋白质指纹图谱模型对RA血清诊断标志物的筛选。方法 用H4蛋白芯片结合SELDI—TOF—MS测定了141例血清标本的蛋白质指纹图谱，其中RA90例，系统性红斑狼疮（SEE）20例，健康志愿者31名。将筛选出的血清蛋白质指纹图谱作为输入，建立人工神经网络预测模型，用随机抽取的93例标本（RA60例，SEE 13例，健康志愿者20名）作为训练组，进行训练与交叉验证，并用另外测试组48例（RA30例，SEE 7例，健康志愿者11名）的血清标本盲法验证该模型，同时与抗环瓜氨酸肽（抗CCP）抗体检测结果进行比较。结果 利用从训练组得出的基于人工神经网络的血清蛋白质指纹图谱模型，对测试组的48例未知血清进行预测．结果显示．对RA检测的敏感性为90％（27／30），特异性为90．9％（9／11），阳性率为90．2％（37／41），明显高于抗CCP抗体检测结果。结论血清蛋白质指纹图谱可有效筛选RA血清中特异性蛋白标志物，基于人工神经网络的血清蛋白质质谱模型较以往传统方法具有更高的敏感性和特异性。     

应用蛋白质指纹技术筛选强直性脊柱炎血清特异性标志物的临床研究

目的应用表面增强激光解析离子化-飞行时间质谱技术(surface-enhanced laser desorption ioni zation/time of flight mass spectrometry,SELDI-TOF-MS)和蛋白质芯片筛选强直性脊柱炎(ankylosing spondylitis,AS)患者血清特异性标志物,用于疾病诊断、评估及预测病情进展。方法采用SELDI-TOF-MS技术和弱阳离子交换(weak cation exchange)芯片检测2008年4月至2009年1月山西医科大学第二医院风湿免疫科收治的69例AS患者及12名健康对照者、10例类风湿关节炎(rheumatoid arthritis,RA)患者血清蛋白质表达,进一步将AS患者分为活动期与非活动期,中轴关节受累及中轴、外周关节均受累,HLA-B27阳性与阴性组,比较不同分组之间患者血清蛋白质指纹图谱,采用SELDI质谱仪自带的Biomarker Wizard和Biomarker Pattern软件筛选,初步建立疾病诊断预测模型。结果由8085、2640和2932建立的诊断预测模型Ⅰ诊断AS的敏感度为94.23%,特异度为100%。由3677、3880、2539、3159和3242建立的诊断预测模型Ⅱ判断病情活动的敏感度为98.11%,特异度为100%。由4700、8687和18538建立的诊断预测模型Ⅲ预测AS同时有中轴及外周关节受累的敏感度为80.00%,特异度为82.35%。由10259、7972、2048、2154和2954建立的诊断模型Ⅳ区分AS和RA的敏感度为100%(69/69),特异度为100%(10/10)。结论通过SELDI-TOF-MS技术建立的血清蛋白质指纹图谱可以筛选AS患者血清中的特异性蛋白质标志物,有望成为诊断疾病及评估病情的一种初筛平台。     

非小细胞肺癌患者血清蛋白质标记物的检测

目的 检测非小细胞肺癌患者(NSCLC)血清蛋白质,筛选特异的蛋白质标记物,构建用于NSCLC早期诊断的血清蛋白质指纹图谱模型.方法 应用表面增强激光解析电离飞行时间质谱(SELDI-TOF-MS)技术检测235例血清标本的蛋白质质谱,并结合生物信息学方法(支持向量机)分析数据.结果 筛选出3个质荷比(m/z)位于6628,9191和11412的蛋白质标记物,构建NSCLC早期诊断模型.联合3种潜在蛋白质标记物,经留一法交叉验证,区分NSCLC和正常健康对照的敏感性为98%,特异性为96%.盲法验证显示,该模型诊断NSCLC的敏感性为96.56%,特异性为94.79%.结论 SELDI-TOF-MS结合支持向量机建立NSCLC血清蛋白质指纹图谱模型是早期诊断NSCLC的一种敏感性高、特异性强的新方法,值得进一步研究与应用.     

应用蛋白质指纹图谱技术快速鉴别诊断菌阴肺结核

目的 探索应用蛋白质指纹图谱技术于菌阴肺结核与肺炎的鉴别诊断。方法 从本院临床病例中,选择菌阴肺结核和肺炎患者及健康者各60例,应用表面加强激光解吸电离飞行时间质谱技术(SELDI/ToF-Ms)和蛋白芯片技术检测血清蛋白,并应用Ciphergen蛋白芯片3.1.1软件进行比较,分析其相关蛋白峰值并进行统计学处理。结果 对180例菌阴肺结核、肺炎患者、健康者的血清蛋白指纹图谱数据进行比较,发现有5个蛋白峰(1 028.49、4 796.56、7 564.77、8 048.02、11 526.75 m/z)存在显著的差异,有统计学意义(P<0.01)。由这5个蛋白峰组成的诊断模型鉴别诊断菌阴肺结核与肺炎的总有效率84.2%(101/120),敏感性与特异性分别为82.5%(52/63),85.9%(49/57);阳性预测值86.7%(52/60),阴性预测值为81.7%(49/60)。诊断模型在判别肺炎、菌阴肺结核患者与健康者之间,总有效率达89.4%(161/180),特异性为100%(60/60),灵敏度为84.2%(101/120),阳性预测值100%(101/101),阴性预测值75.9%(60/79)。结论 蛋白质指纹图谱技术具有方法简便、检测快速,标本用量少的优点,是筛选结核病特异性标志物的有效手段,通过蛋白质指纹图谱技术检测,发现了具有良好鉴别诊断的“诊断模型”。     

胃癌与慢性胃炎唾液蛋白质组鉴别诊断模型

目的:探讨用蛋白质组学质谱技术筛选慢性胃炎患者与胃癌患者唾液蛋白质表达谱,寻找可用于胃癌与慢性胃炎鉴别诊断的特异性生物标志物.方法:采用蛋白质组学基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术,并运用弱阳离子磁珠(WCX)检测慢性胃炎患者和胃癌患者的唾液,得到相应的肽质量指纹图谱,建立鉴别诊断模型.结果:慢性胃炎患者与胃癌患者两组共得到蛋白质峰为77个,其中1个有统计差异显著的蛋白峰(P<0.05),质荷比为:6021.72 Da,通过分析差异蛋白峰表达谱,建立了分类预测模型,识别率为83.54%,预测能力60.23%.以此模型进行临床回代检验结果14例慢性胃炎,其中10例被准确检出,23例胃癌,22例被准确检出,灵敏度71.43%(10/14),特异度95.65%(22/23).结论:初步得到了慢性胃炎与胃癌唾液差异蛋白质表达谱,并初步建立了以6021.72 Da蛋白质区分慢性胃炎与胃癌的唾液蛋白鉴别诊断模型.     

The immunoneuroendocrine role of melatonin

Abstract: A tight, physiological link between the pineal gland and the immune system is emerging from a series of experimental studies. This link might reflect the evolutionary connection between self-recognition and reproduction. Pinealectomy or other experimental methods which inhibit melatonin synthesis and secretion induce a state of immunodepression which is counteracted by melatonin. In general, melatonin seems to have an immunoenhancing effect that is particularly apparent in immunodepressive states. The negative effect of acute stress or immunosuppressive pharmacological treatments on various immune parameters are counteracted by melatonin. It seems important to note that one of the main targets of melatonin is the thymus, i.e., the central organ of the immune system. The clinical use of melatonin as an immunotherapeutic agent seems promising in primary and secondary immunodeficiencies as well as in cancer immunotherapy. The immunoenhancing action of melatonin seems to be mediated by T-helper cell-derived opioid peptides as well as by lymphokines and, perhaps, by pituitary hormones. Melatonin-induced-immuno-opioids (MHO) and lymphokines imply the presence of specific binding sites or melatonin receptors on cells of the immune system. On the other hand, lymphokines such as -γ-interferon and interleukin-2 as well as thymic hormones can modulate the synthesis of melatonin in the pineal gland. The pineal gland might thus be viewed as the crux of a sophisticated immunoneuroendocrine network which functions as an unconscious, diffuse sensory organ.     

Response of rat pineal melatonin to calcium, magnesium, and lithium is circadian stage dependent

Abstract: Herein we documented the response of pineal melatonin production to electrolytes known to be effective on pineal function in view of a possible circadian stage dependence. We studied the release of melatonin by perifused rat pineal glands at 2 different circadian stages corresponding to the middle of the light and dark periods, i.e., respectively, 7 and 19 HALO (Hours After Light Onset, L:D = 12:12). The initial efflux rates were, as expected, much higher in the perifusates of glands removed from rats sacrificed during the dark phase than of those removed during the light phase. After 3 hr of perifusion, melatonin release reached similar levels which were found constant up to the 8th hr of perifusion, whatever the circadian stage. Perifusion of the glands with physiological concentrations for the rat of calcium (5.2 mmol/1) and magnesium (1.34 mmol/1) resulted in a stimulatory effect on the pineal glands removed from rats sacrificed in the middle of the dark period (19 HALO), whereas no effects were observed on the pineal glands removed from rats sacrificed during the light (7 HALO). Lithium (0.28 and 0.55 mmol/1) was ineffective on melatonin release in pineal glands removed 7 and 19 HALO. Our results show differences in the initial efflux rates of melatonin and in the response of perifused pineal glands to calcium and magnesium according to the circadian stage.     

Changes in connexin43, the gap junction protein of astrocytes, during development of the rat pineal gland

Abstract: The abundance of gap junctions between rat pineal astrocytes formed by connexin43 (Cx43) was studied during development. Levels and distribution of Cx43 were measured by immunoblotting and indirect immunofluorescence, respectively. The amount of Cx43 in cells located within the gland was low until about the 7th postnatal day and increased to adult values between the 14th and 21st days postpartum. Although astrocytes, recognized by their vimentin immunoreactivity, were scarce before birth, they were abundant by the 7th postnatal day suggesting that the low levels of Cx43 found at this age corresponded to a low expression of this protein. Localization of the immunoreactivity to Cx43 and vimentin showed a close correlation, indicating that mature or immature pineal astrocytes form gap junctions made of Cx43. Since Cx43 levels attained their adult values at about the time the innervation and the functional state of the gland reached maturity (2–3 weeks after birth), it is proposed that astrocyte gap junctions are involved in the function of the adult rat pineal gland.     

Conservative management of perforated duodenal diverticulum： A case report and review of the literature

Duodenal diverticula are a relatively common condition. They are asymptomatic, unless they become complicated, with perforation being the rarest but most severe complication. Surgical treatment is the most frequently performed approach. We report the case of a patient with a perforated duodenal diverticulum, which was diagnosed early and treated conservatively with antibiotics and percutaneous drainage of secondary retroperitoneal abscesses. We suggest this method could be an acceptable option for the management of similar cases, provided that the patient is in good general condition and without septic signs.     

Presence of immunoreactive growth hormone and prolactin in the ovine pineal gland

Abstract: The use of antisera raised against bovine growth hormone (GH) and ovine prolactin (PRL) enabled the detection of related immunoreactive (ir) sequences of proteins in ovine pineal tissue. The isolation of PRL-like ir-material was accomplished using a 0.25 M ammonium sulphate (pH 5.5) extraction followed by ethanol precipitation, whereas the resulting 2.0 M ammonium sulphate (pH 7.0) precipitate contained a GH-like immunoreactivity. Gel chromatography of the GH-like immunoreactivity (Sephadex G-100) indicated the presence of several GH-like fragments ranging in the M_r range of 7,000 to 55,000. Analyses of the PRL-like ir-material found in pineal tissue on HPLC using a TSK 545-DEAE column led to the resolution into a single peak of immunoreactivity. A single peak of activity was also observed following chromatofocusing and hydrophobic interaction chromatography of the ir-peak from the TSK 545-DEAE column. The PRL-like ir-material inhibited the binding of [¹²⁵I]ovine PRL-S14 to anti-ovine PRL antibodies without showing an affinity for binding to anti-rat PRL or anti-bovine GH antibodies. Scatchard analysis of the binding of pineal PRL-like ir-material and pituitary ovine PRL-S14 to liver membranes from day-20 pregnant rats revealed similar affinity constants (K_a of 4.7 ± 0.2 × 10⁹ M^-1). In addition, the replication of Nb 2 Node rat lymphoma cells was stimulated by pineal PRL-like ir-material, an effect known to be specific for lactogenic hormones. The pineal PRL-like immunoreactivity appeared on sodium dodecyl sulfate polyacrylamide gels as a single major band of M_r 24,000. The functional status of PRL-and GH-like ir-material in the ovine pineal remains to be determined, but evidence is presented that the overall protein synthesis rate of the rat pineal responded to circulating concentrations of PRL.     

Microbiology of human immunodeficiency virus anorectal disease

PURPOSE: Individuals who are seropositive for the human immunodeficiency virus are at high risk for opportunistic infection and anorectal disorders. Little prospective information is available regarding anorectal pathogens in these patients. METHODS: One hundred sixty-three HIV-seropositive patients presented to the colorectal clinic between 1989 and 1992. Forty-seven (29 percent) patients were thought to have an infectious process and were prospectively studied using a standardized multiculture protocol. RESULTS: Mean age was 33 (range, 19–59) years. All were male; high-risk behavior accounted for 87 percent of HIV transmissions. Presenting complaints included anorectal pain (79 percent), pus per anum (28 percent), and blood per anum (26 percent). Examination revealed perianal tenderness (60 percent), condyloma (38 percent), perianal ulcers (38 percent), and anal fissures (34 percent). Sixty-six sets of cultures were performed; 28 patients had one set, 15 had two sets, and 4 had three sets. Thirty-two of these 47 patients (68 percent) had positive cultures including herpes (50 percent), cytomegalovirus (25 percent),Neisseria gonorrhoeae (16 percent), chlamydia (16 percent), acidfast bacilli (2 percent), and others (9 percent). Six of 32 patients with positive cultures had more than one organism cultured. Sixteen (50 percent) patients with positive cultures were treated medically, 8 (25 percent) were treated surgically and 8 (25 percent) were treated with both modalities. Sixty-one procedures were performed on 17 patients for condylomata. Eighteen patients had 20 procedures for abscesses, 50 percent of whom had positive cultures for other than common bowel flora; all improved. Fourteen patients underwent 33 procedures for perianal fistulas.Mycobacterium fortuitum was cultured from one patient who required 13 procedures for abscesses and fistulas. Forty-five (96 percent) patients were followed for an average of 12.5 months ±2.9 SEM (range, 1–94 months). Symptoms were improved or resolved in 22 of 32 (69 percent) patients with positive cultures and in 11 of 13 (84 percent) with negative cultures. CONCLUSIONS: Specific pathogens may often be identified in human immunodeficiency virus-seropositive patients with anorectal disorders if aggressively sought. Although patients without specific pathogens identified may be expected to improve with planned empiric treatment, positive identification allows more directed therapy.