洲滩草本植物对日本血吸虫毛蚴杀灭作用的观察

分别用10%的青蒿(茎)、猫眼草(茎)、毛茛(叶)、藜蒿(根)和益母草浸出液对日本血吸虫毛蚴进行杀灭试验,作用1min后的毛蚴死亡率分别为88.28%、84.54%、78.42%、62.62%和68.40%。表明以上植物对毛蚴有一定杀灭作用。     

细粒棘球绦虫转基因植物载体重组pBI-Eg95质粒构建及鉴定

目的 构建并鉴定细粒棘球绦虫(Eg)转基因植物载体重组pBI-Eg95质粒.方法 从细粒棘球蚴包囊中分离原头节.经超声粉碎后抽提总RNA,反转录成eDNA,设计合成引物,以eDNA为模板.通过PCR从cDNA中扩增出目的 基因Eg95,经电泳及测序鉴定后,将该基因定向克隆到植物表达载体pBI121中构建pBI-Eg95重组质粒,电穿孔转化根癌农杆菌(At)LBA4404株;从转化的At阳性株中抽提质粒进行双酶切和以抽提的质粒为模板进行PCR鉴定.结果 RT-PCR扩增出1条约471 bp的特异性条带,DNA序列分析与GenBank知的序列同源性为100%.从转化的At中抽提的质粒,双酶切及PCR测定的结果与预期相符.结论 成功构建了细粒棘球绦虫转基因植物载体重组pBI-Eg95质粒,为进一步构建细粒棘球绦虫转基因植物疫苗奠定了基础.     

Demographic,clinical and allergological characteristics of Eosinophilic Esophagitis in a Spanish central region

BackgroundEosinophilic esophagitis (EoE) is a chronic inflammatory emerging disease of the oesophagus with immunoallergic aetiology. The allergens involved have not been clearly defined and may depend on the exposure of the population to aeroallergens or food antigens.Materials and methodsPatients diagnosed with EoE between 2006 and 2011 were referred to our Allergy Section. Patch and skin prick tests (SPT) with aeroallergens and foods were performed, and total and specific IgE levels, eosinophil cationic protein levels and eosinophil count were determined.Results43 patients were included. 36 (83.7%) were atopic. 29 patients presented choking, 19 dysphagia, 9 food impaction with urgent endoscopy, 4 chest pain, 1 isolated vomiting and 1 epigastric pain. 22 had two or more symptoms. The mean duration of symptoms was 3.73 years. Concomitant allergic diseases included rhinoconjunctivitis and/or asthma (31 patients), IgE food allergy (21 patients) and atopic dermatitis (3 patients).32 (74%) were sensitized to aeroallergens, of which 90% were sensitized to pollens; 23 (54%) showed positive tests to foods and 12 of them (52%) to lipid transfer proteins (LTP).Of the 29 pollen-allergic patients, 15 (52%) were sensitized to plant foods and 10 (34.4%) to LTP.ConclusionsOur findings support those reported in the literature: the disease is more common in men aged 30–40 years with at least a three-year history of symptoms of esophageal dysfunction, sensitized to pollens, the predominant aeroallergen in our area, but also to plant foods or panallergens. These results increase the evidence for an immunoallergic aetiology and can help us in the early diagnosis of EoE.     

Bactericidal and anti-adhesive properties of culinary and medicinal plants against Helicobacter pylori

AIM: To investigate the bactericidal and anti-adhesive properties of 25 plants against Helicobacter pylori (H pylori). METHODS: Twenty-five plants were boiled in water to produce aqueous extracts that simulate the effect of cooking. The bactericidal activity of the extracts was assessed by a standard kill-curve with seven strains of H pylori. The anti-adhesive property was assessed by the inhibition of binding of four strains of FITC-labeled H pylori to stomach sections. RESULTS: Of all the plants tested, eight plants, including Bengal quince, nightshade, garlic, dill, black pepper, coriander, fenugreek and black tea, were found to have no bactericidal effect on any of the isolates. Columbo weed, long pepper, parsley, tarragon, nutmeg, yellow-berried nightshade, threadstem carpetweed, sage and cinnamon had bactericidal activities against H pylori, but total inhibition of growth was not achieved in this study. Among the plants that killed H pylori, turmeric was the most efficient, followed by cumin, ginger, chilli, borage, black caraway, oregano and liquorice. Moreover, extracts of turmeric, borage and parsley were able to inhibit the adhesion of H pylori strains to the stomach sections. CONCLUSION: Several plants that were tested in our study had bactericidal and/or anti-adhesive effects on H pylori. Ingestion of the plants with anti-adhesive properties could therefore provide a potent alternative therapy for H pylori infection, which overcomes the problem of resistance associated with current antibiotic treatment.     

细粒棘球绦虫转Eg95-EgA31融合基因苜蓿疫苗免疫小鼠后脾细胞因子的动态观察

目的 动态观察细牲棘球绦虫(Eg)转Eg95-EgA31融合基因苜蓿疫苗免疫Balb/c小鼠后脾细胞因子的变化.方法 88只Balb/c小鼠按体质量随机分为两组,用转基因苜蓿叶蛋白提取液(20 s/L)100 μl口服灌胃和10μl鼻腔黏膜接种分别免疫小鼠,每3天1次,连续2个月.在末次免疫后0(对照)、2、4、6、8、10、12、14、16、18、20周各组剖杀4只小鼠,取脾,分离脾细胞,体外经Eg粗抗原(EgAg)或刀豆蛋白A(ConA)刺激培养48 h,诱生白细胞介素(IL)-12、γ干扰素(IFN-γ)和IL-10;经EgAg或脂多糖(LPS)刺激培养72 h,诱生肿瘤坏死因子α(TNF-α).收集脾细胞培养上清液,常规ELISA法检测IL-12、IFN-γ、TNF-α和IL-10水平.结果 口服灌胃组的IL-12、IFN-γ、TNF-α和IL-10水平分别在免疫后4～6、2～8、2～6、4～12周升高,分别在免疫后4、2、2、8周达最高水平,其值分别为(25.0±5.8)、(575.0±28.9)、(50.0±11.5)、(42.5±2.9)ng/L,与0周[(11.3±2.5)、(125.0±28.9)、(11.3±2.5)、(12.5±2.9)ng/L]比较,差异有统计学意义(P均<0.01);鼻腔黏膜接种组的IL-12、IFN-γ、TNF-α和IL-10水平分别在免疫后4～6、2～10、4～10、6～16周升高,分别在免疫后6、4、6、6周达最高水平,其值分别为(25.0±5.8)、(725.0±28.9)、(27.5±2.9)、(60.0±11.5)ng/L,与0周[(11.3±2.5)、(125.0±28.9)、(11.3±2.5)、(12.5±2.9)ng/L]比较,差异有统计学意义(P均<0.01).EgAg、ConA、LPS刺激组的细胞因子水平高于相应的脾细胞悬液组(P<0.05或<0.01),ConA或LPS刺激组的细胞因子水平高于相应的EgAg刺激组(P<0.05或<0.01).结论 细粒棘球绦虫转Eg95-EgA31融合基因苜蓿疫苗在免疫早期(2～10周)可诱导小鼠产生Th1和Th2混合型免疫应答.     

细粒棘球绦虫转Eg95基因苜蓿疫苗的培育及鉴定

目的 培育并鉴定细粒棘球绦虫转Eg95基因苜蓿疫苗.方法 利用转pBI-Eg95质粒的根癌农杆菌LBA4404株介导的苜蓿子叶浸染法,将Eg95基因导人紫花苜蓿基因组,转Eg95基因苜蓿外植体在含有卡那霉素的选择培养基上经愈伤、出芽和生根阶段生长出小苗,最后移栽到装有营养土的花盆中,生长2～3个月,获得完整的转Eg95基因苜蓿疫苗.提取转Eg95基因苜蓿的DNA、RNA及叶蛋白,采用PCR、RT-PCR、十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)及Western blot法进行鉴定.结果 PCR、RT-PCR法在471 bp处均扩增出目的条带;SDS-PAGE及Western blot法可见转Eg95基因苜蓿蛋白在相对分子质量约16.5×103处出现特异条带,与预期结果相符;Bio-Rad Quantity one系统分析表达效率约占提取总苜蓿叶蛋白的0.06%.结论 成功培育出细粒棘球绦虫转Eg95基因苜蓿疫苗.     

细粒棘球绦虫转Eg95-EgA31融合基因苜蓿疫苗诱导小鼠脾细胞亚群的变化

目的 探讨细粒棘球绦虫(Eg)转Eg95-EgA31融合基因苜蓿疫苗免疫和Eg原头节攻击后小鼠脾CD4+和CD8+T淋巴细胞亚群的变化.方法 热絮凝法提取转基因苜蓿叶蛋白,用无菌双蒸水将叶蛋白提取液配成20 g/L,同时提取转空质粒(pBI121)苜蓿叶蛋白及正常苜蓿叶蛋白作对照.32只雌性Balb/c小鼠按体质量随机分为4组,每组8只.口服灌胃组:灌胃接种100 μl转基因苜蓿叶蛋白提取液;鼻腔黏膜接种组:滴鼻接种10 μl转基因苜蓿叶蛋白提取液;空质粒对照组:滴鼻接种10μl转空质粒苜蓿叶蛋白提取液;正常蛋白对照组:灌胃接种100μl正常苜蓿叶蛋白提取液.小鼠每3天免疫1次,连续免疫2个月.末次免疫后第8周,各组小鼠用Eg原头节腹腔注射攻击感染(50个/只),感染后第24周剖杀小鼠,分离脾细胞,用流式细胞仪检测脾CD4+和CD8+T淋巴细胞亚群百分比.结果 口服灌胃组小鼠脾CD4+T细胞亚群百分比(0.286±0.009)、CD8+ T细胞亚群百分比(0.102±0.004)和CD4+/CD8+比值(2.814±0.014)均显著高于正常蛋白对照组(0.166±0.018、0.083±0.006、2.019±0.369,P<0.01或<0.05);鼻腔黏膜接种组小鼠脾CD4+T细胞亚群百分比(0.269±0.016)和CD4+/CD8+比值(2.955±0.986)与正常蛋白对照组比较明显升高(P均<0.01);口服灌胃组小鼠脾CD4+T细胞亚群百分比高于鼻腔黏膜接种组(P<0.05);空质粒对照组小鼠脾CD4+、CD8+T细胞百分比和CD4+/CD8+比值(0.169±0.018、0.093±0.019、1.852±0.188)与正常蛋白对照组比较,差异无统计学意义(P均>0.05).结论 CD4+T细胞亚群可能与细粒棘球绦虫转Eg95-Eg31融合基因苜蓿疫苗诱导的小鼠抗Eg原头节攻击感染的保护力有关.疫苗灌胃接种可能是一种较好的免疫途径.     

提高对“土三七”致肝小静脉闭塞病的认识

我国肝小静脉闭塞病（HVOD）的病因多见于长期或过量服用土三七。土三七内舍有吡咯烷生物碱成分,可造成肝窦和肝小静脉的内皮细胞损伤．导致肝小静脉阻塞。HVOD所致的血流阻力部位为肝内,引起窦后性门静脉高压。肝大、右上腹痛、腹水和黄疸是其主要临床表现。诊断依据：有长期服用或过量服用土三七史;有上述临床表现;血清丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）、1-谷氨酰转肽酶（GGT）、碱性磷酸酶（ALP）、胆红素（BiL）均增高;B超可见腹水、肝肿大、肝区呈“斑片状”低回声区,肝静脉显示不清或不显示;CT检查可见肝肿大、腹水,增强CT动脉期显示全肝密度不均匀,呈现“斑片状”或“地图状”改变。肝活检早期可见肝小静脉管腔狭窄,肝小静脉周围血管阻塞．肝细胞不同程度液化坏死．晚期可见纤维化。早期诊断和及时治疗有较好的预后。临床医师应提高对HVOD的认识．且要提醒人们不要滥用土三七。     

Lack of herbal supplement characterization in published randomized controlled trials

PURPOSE: Herbal supplements in the United States and abroad have poor quality control and high content variability. We assessed the extent to which recently published randomized controlled trials of herbal supplements characterized and verified the content of the supplement under study. METHODS: We identified all MEDLINE-indexed English language randomized controlled trials evaluating single-herb preparations of echinacea, garlic, ginkgo, saw palmetto, or St. John's wort that were published between January 1, 2000, and February 9, 2004. From each article we extracted information characterizing the herbal supplement studied. RESULTS: Of 81 randomized controlled trials meeting inclusion criteria, 12 (15%) reported performing tests to quantify actual contents, and 3 (4%) provided adequate data to compare actual with expected content values of at least one chemical constituent. In those 3 studies, actual content varied between 80% and 113% of expected values. Studies of higher overall quality (Jadad score > or =3) performed testing somewhat less frequently (5/54; 9%) than those with lower Jadad scores (7/27; 26%) (P = .09). CONCLUSION: Documented characterization of herbal supplements in published randomized controlled trials is inadequate. Investigators may be unaware of the extent to which herbal quality-control issues may detract from the value of otherwise well-designed clinical trials. The scientific and clinical utility of future herbal randomized controlled trials would be enhanced if authors provided evidence that the herbal products studied were of high quality.     

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.     

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.     

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.     

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.