湖北贝母的HPLC指纹图谱分析

目的建立湖北贝母的指纹图谱分析方法。方法采用高效液相色谱-蒸发光散射检测法以Hypersil ODS(250 mm×4.6 mm ID，5 μm)为色谱柱；流动相：甲醇(含0.05%三乙胺)-水梯度洗脱；流速：1.0 mL·min^-1；记录时间：60 min；以蒸发光散射检测器(ELSD)检测，检测条件：漂移管温度75 ℃，氮气流速1.9 L·min^-1。结果用梯度洗脱得到的色谱图各色谱峰分离较好，达到指纹图谱要求。结论为更好的控制湖北贝母的内在质量提供了可靠的分析方法。     

Allergenic epitopes of bovine αS1-casein recognized by human IgE and IgG

B-cell epitopes of bovine α_S1-casein, one of the major allergens of cow's milk, were identified by a screening method based on synthetic peptides. According to the known amino acid sequence of α_S1-casein, a set of 188 overlapping sequential decapeptides shifted by one amino acid was manually synthesized on polyethylene pins by the 9-fluorenyl-methoxycarbonyl (Fmoc) method. Peptides were screened by an enzyme-linked immunosorbent assay (ELISA) specific for human IgE and IgG. Bound antibodies were detected by successive incubation with up to three polyclonal antibodies, the last one conjugated to horseradish peroxidase. Tested sera were from 15 patients with acute clinical reactions to cow's milk and IgE-specific reactions to bovine α-casein in the ELISA and immunoblot. Sera from 10 healthy subjects without remarkable reactions to cow's milk proteins were used as controls. All sera from allergic subjects showed reactions with three regions of α_S1-casein, corresponding to amino acids 19–30, 93–98, and 141–150. Furthermore, individual sera showed reactions with other parts of the protein. No essential differences in the epitope specificity of IgE and IgG were found. Inhibition of IgE binding to α_S1-casein with soluble synthetic peptides confirmed the results and revealed peptide CN-2 as the most inhibiting one.     

The attentional selection of spatial and non-spatial attributes in touch: ERP evidence for parallel and independent processes

To investigate the functional relationship between spatial and non-spatial attentional selectivity in somatosensory processing, event-related potentials (ERPs) were recorded to mechanical tactile stimuli, which were delivered to the right or left hand, and were low or high in frequency (Experiment 1), or soft or strong in intensity (Experiment 2). Participants' task was to attend to a specific combination of one stimulus location and one non-spatial attribute. Spatial attention was reflected in enhanced N140 components followed by a sustained attentional negativity. ERP effects of non-spatial attention (enhanced negativities to the attended frequency or intensity) were observed in the same latency range, suggesting that the attentional selection of relevant spatial and non-spatial attributes occurs in parallel. Most importantly, ERP correlates of attention directed to stimulus frequency and intensity were unaffected by the current focus of spatial attention. In contrast to vision, where the selective processing of non-spatial attributes is hierarchically dependent on selection by location, but similar to auditory attention, spatial and non-spatial attentional selectivity appear to operate independently in touch.     

Role of angiotensin Ⅱ and JAK2 signal pathway in transdifferentation of renal tubular cells in mice after acute ischemic followed by reperfusion

Objective To investigate the effect of angiotensin (Ang) Ⅱ and its Janns-activated kinase-2 (JAK2) signal pathway in transdifferentiation of renal tubular cells under the challenge of acute ischemic reperfusion injury.Methods Models of acute ischemic reperfusion injury were established and the level of local Ang Ⅱ ,a key element of renin-angiotensin system (RAS),in kidney was measured using radioimmunity technique.The expression of α-smooth muscle actin (α-SMA),a phenotype of mesenchymal cells,was detected by RT-PCR and inununohistochemistry methods.Renal tubule cells ( NRK-52E) were cultured with various concentration of Ang Ⅱ ,followed by blocking of PD123319,Ang U receptor 2 antagonist,and AG490,an inhibitor of JAK2 signal pathway.Results Ang 0 of kidney tissue increased immediately after acute ischemic-reperfusion injury,in time dependent fashion.Expression of α-SMA in renal tubule cells was found at 48 hours after ischemic-reperfusion injury and in NRK-52E cells treated by high concentration of Ang Ⅱ and was dose and time dependent.The peak of α-SMA expression was seen after 30 minute treatment at the dose of 10-9'mol/L,which was interrupted by both of PD123319 and AG490.Conclusions Transdifferentiarion of renal tubular epithelial cells occurs under acute ischemic- reperfusion injury.Local renin-angiotensin system may play a role in the transdifferentiation of TEC through AT2 receptor and its JAK2 signal pathway.     

姜黄素抑制肝纤维化大鼠脂质过氧化物的生成及肝脏TGF-β1和PDGF的表达

目的： 观察姜黄素对肝纤维化过程中活性氧（ROS）、丙二醛（MDA）的生成及转化生长因子-β₁（TGF-β₁）、血小板源性生长因子（PDGF）表达的影响，探讨姜黄素预防肝纤维化的作用机制。方法: 采用四氯化碳腹腔注射方法，每周3次，共8周制作大鼠肝纤维化模型，同时每只大鼠按每100 g体重分别给予20 mg、10 mg、5 mg姜黄素灌胃处理，每周3次，共8周；设立正常组、肝纤维化组和阳性对照组。8周后处死大鼠，留取大鼠肝脏和血清。比色法检测血清ROS水平；硫代巴比妥酸（TBA）法检测肝组织匀浆中MDA含量；免疫组化方法检测各组大鼠肝组织中TGF-β₁、PDGF的表达水平。结果: 模型组大鼠血清ROS水平明显升高，姜黄素可显著降低ROS水平（P＜0.05）；姜黄素组大鼠肝组织中MDA含量明显低于模型组（P＜0.01）；模型组大鼠肝组织中TGF-β₁、PDGF大量表达，姜黄素可明显抑制上述因子的表达（P＜0.01）。结论: 姜黄素可抑制肝脏脂质过氧化物的形成以及TGF-β₁和PDGF的表达，从而发挥预防肝纤维化的作用。     

Effects of vitamin E on human platelet and mononuclear cell responses in vitro

Recent epidemiological studies have provided evidence supporting the potential benefits of antioxidants in coronary prevention. We have investigated the effects of vitamin E on platelets, monocytes and endothelial cells in vitro. Pre-incubation of platelets with vitamin E inhibited subsequent thrombin- (P < 0.05, n = 5), collagen- (P < 0. 0001, n = 5) and ADP-(P < 0.05, n = 4) induced platelet aggregation measured using a microtitre plate method, or conventional aggregometry. The adhesion of thrombin-activated platelets to collagen was also inhibited by vitamin E (P < 0.05, n = 8), but not by vitamin C (P > 0.05, n = 8); nor was the adhesion of unstimulated platelets significantly affected (P > 0.05, n = 8). Pre-incubation of monocytes with vitamin E inhibited their subsequent adhesion to plastic (P < 0.05, n = 9), and was also associated with an 18% reduction in adhesion to EA.hy 926 endothelial cells (n = 8), although this failed to reach statistical significance. Pre-incubation of the endothelial cells with vitamin E also significantly reduced subsequent mononuclear cell adhesion by 56% (P < 0.05, n = 3).     

Feasibility study of a primary care-based model for stroke aftercare

BACKGROUND: A new primary care model for stroke aftercare has been developed to address the longer-term needs of patients who have had a stroke and their families. AIM: To test the feasibility and acceptability of the new model on patients who have had a stroke and their carers. DESIGN OF STUDY: An observational feasibility study. SETTING: Patient and carer assessments and interviews conducted in patients' homes. METHOD: Patients who have had a stroke, and their carers, were assessed using a purposely developed primary care stroke model. Qualified health- and social-care professionals acted as stroke care coordinators and carried out the assessments. Four evaluations were undertaken as part of the study: analysis of care plans, a survey of unmet needs, focus group sessions for staff, and patient/carer interviews. RESULTS: Forty-seven patients and 21 carers were recruited to the study. Analysis of care plans indicated that the assessment process was successful in identifying patient and carer problems (n = 219). Actions were instigated against 190 of these problems and, at 3 months after the assessment, 75% of the problems had been resolved. Patients/carers thought that the review process would be more valuable if conducted sooner after hospital discharge. CONCLUSION: The model was acceptable to professionals, encouraging them to work in a manner more consistent with the expressed needs of patients and carers who were adjusting to the longer-term impact of stroke. Essential aspects informing the care process were also identified. Further work is now required to investigate the effects of the intervention on patient and carer outcomes.     

Enzyme-linked immunosorbent assay of allergen-specific IgG antibodies in bee sting allergic patients hyposensitized with pure bee venom

Summary An ELISA is presented for detection of IgG antibodies to bee venom. By this method, sera of 11 bee sting allergic patients, who were treated with rapid hyposensitization with pure bee venom, were tested. The highest antibody titers were observed after 30 days of treatment, a maximum rise of 7.4±1.5 log 2-titer steps. Pure bee venom is shown to be more potent immunologically than whole body bee extract. Prediction of the clinical success, measured by tolerance to a bee sting challenge, is not yet possible using venom specific IgG determinations.This work was supported by the Deutsche Forschungsgemeinschaft, grant UR 12/2     

The recognition pattern of sequential B cell epitopes of beta-lactoglobulin does not vary with the clinical manifestations of cow's milk allergy

BACKGROUND: beta-Lactoglobulin (BLG) represents one of the major allergens causing cow's milk allergy (CMA) - a disease with a wide spectrum of clinical symptoms. The aim of this study was to evaluate sequential B cell epitopes of BLG by the Pin-ELISA method. Furthermore, we wanted to investigate a possible association of the IgE recognition patterns in sera of patients with BLG sensitization and the type of clinical reactions following contact with cow's milk. METHODS: Overlapping sequential decapeptides corresponding to the amino acid sequence of BLG were used in Pin-ELISAs specific for human IgE. Tested sera were from 14 individuals with CMA, 8 of them with a history of immediate systemic reactions and 6 with delayed skin reactions following contact with cow's milk. All of them showed specific IgE antibodies to BLG in the CAP-RAST. Control sera were from 5 healthy nonallergic individuals. RESULTS: All sera from BLG-sensitized individuals showed IgE binding with one region of BLG corresponding to amino acids 95-113. Furthermore, individual sera showed reactions with two further regions, 12-27 and 124-135. Inhibition of IgE binding to BLG with one soluble synthetic peptide confirmed the major epitope. No differences were found in the B cell epitope recognition pattern to BLG in the two groups of patients with CMA, characterized by acute systemic or delayed skin reactions. CONCLUSIONS: Using IgE Pin-ELISAs we were able to confirm previously described sequential B cell epitopes of BLG. However, the recognition pattern of one of the major cow's milk allergens is not predictive of the clinical type of reaction.