Structural characterization of the C4a anaphylatoxin from rat

The C4a anaphylatoxin was purified from rat sera activated by heat-aggregated IgG. The anaphylatoxin was isolated by a three-step purification procedure and was judged to be homogeneous based on visualization of a single stained band after electrophoresis on both cellulose acetate membrane strips and on 9% SDS-polyacrylamide gels. Results from Ouchterlony and radioimmunoassay analysis indicated that neither rat C5A nor C3a contaminated the C4a preparation. Rat C4a is a glycoprotein estimated to be 11,000-12,000 mol. wt and contains 76 amino acid residues representing a mol. wt of 8577 and one oligosaccharide unit of 2000-3000 mol. wt. Rat C4a is weakly active in contracting guinea pig ileum at 0.1-1 microM, which is comparable with the activity of human C4a. Both human and bovine C4a are polypeptides free of carbohydrate while rat and presumably mouse C4a are glycoproteins. The complete primary structure of rat C4a anaphylatoxin has been elucidated as follows: (formula; see text)     

人肝癌组织中p53与HSP70相互作用的初步研究

目的 :探讨人肝癌中热休克蛋白 70 (HSP70 )与 p5 3的相互作用。方法 :用免疫组织化学染色法 ,从 12例肝癌组织中筛选HSP70与 p5 3均呈阳性表达的标本 ,并以免疫共沉淀法提取之。然后用SDS PAGE及Westernblot分析双阳性标本中两种蛋白的存在形式。结果 :用免疫组织化学法检测到 12例肝癌组织中有 3例为双阳性 ,用抗HSP70mAb免疫共沉淀的样品 ,可检测到 p5 3蛋白。用抗p5 3mAb免疫共沉淀的样品也可检测到HSP70蛋白。结论 :人肝癌中p5 3与HSP70以复合物的形式而存在 ,此可为肝癌的发病机制及免疫治疗的研究提供新的思路     

CD54和LFA-1的相互作用在6A8α-甘露糖苷酶表达抑制的JurkatT细胞的黏附性增强中的作用

目的研究CD54和LFA-1的相互作用在6A8 α-甘露糖苷酶表达抑制的Jurkat细胞(AS)的黏附性增强中的作用。方法用细胞凝集试验确证AS细胞间黏附的增强，用细胞与细胞间黏附分子-1-人IgG的Fc片段(ICAM-1-Fc)的黏附试验和阻断性抗CD11a抗体的阻断试验研究CD54-LFA-1的作用，用单克隆抗体MEM-148检测As细胞LFA-1亲和力的变化，用单克隆抗体NKI-L16检测AS细胞LFA-1亲合力的变化，用鬼笔环肽染色细胞骨架，用Jurkat-Raji细胞间的作用作模型研究6A8α-甘露糖苷酶表达抑制对T和B细胞间黏附的影响，用ConA结合试验检测细胞中蛋白质N-糖基化的变化。结果(1)AS细胞间的黏附性增强主要与CD54及CD11a表达的增强相关，也与LFA-1亲和力的增高相关；(2)AS细胞的细胞骨架发生重排；(3)As细胞与Raji细胞间的黏附也增强；(4)ConA与AS细胞的结合增强。结论CD54和LFA-1的相互作用在AS Jurkat T细胞的黏附性增强中起重要作用。细胞骨架重排也可能起作用。As细胞的蛋白质发生了N-糖基化修饰。     

Deletion of stem-loop 3 is compensated by second-site mutations within the Gag protein of human immunodeficiency virus type 1

Encapsidation of human immunodeficiency virus type 1 (HIV-1) RNA involves specific interactions between viral Gag proteins and viral RNA elements located at the 5' untranslated region (UTR). These RNA elements are termed packaging (psi) or encapsidation (E) signals and mainly comprise the stem-loop 1 (SL1) and SL3 RNA structures. We have previously shown that deletion of the SL1 sequences is compensated by second-site mutations within Gag. Similar studies are now extended to SL3 and the results demonstrate that deletion of this RNA structure is rescued by two point mutations, i.e., A11V in p2 and I12V in nucleocapsid (NC). These two compensatory mutations are different from those associated with the rescue of SL1 deletion, suggesting that SL1 and SL3 may bind to different residues of Gag during viral RNA packaging. Analysis of virion-derived RNA in native agarose gels shows that deletion of SL3 leads to decreases in both viral RNA packaging and dimerization. These defects are corrected by the compensatory mutations A11V and I12V. Yet, defects in viral RNA dimerization at an early stage that were caused by the SL3 deletion in the context of a viral protease-negative mutation cannot be overcome by these two suppressor mutations. Therefore, the positive effects of A11V and I12V on dimerization of the SL3-deleted RNA must have taken place at the maturation stage.     

原发性高血压患者红细胞[Ca2+]i浓度变化及影响因素

目的：观察原发性高血压患者红细胞[Ca^2 ]i,多巴胺β羟化酶及ATP含量变化并分析其结果。方法：测定35例高血压患者的红细[Ca^2 ]i、ATP、血清多巴胺β羟化酶活性,血糖及血浆胰岛素含量,并以30例健康成年人为对照。结果：高血压患者的红细胞[Ca^2 ]i、ATP、多巴胺β羟化酶均明显高于对照组（P＜0．05,P＜0．01）,但血糖与胰岛素未见明显变化。结论：高血压患者血清多巴胺β羟化酶活性增强伴随ATP与[Ca^2 ]i升高。     

Analysis of pharmacological mechanisms of Yinyanghuo as treatment of erectile dysfunction with network pharmacology-based strategy

Erectile dysfunction is considered an important health problem that impacts the quality of life of men. Yinyanghuo, also called Epimedium or Horny Goat Weed, is a frequently used Chinese traditional herbal medicine, commonly used in treating erectile dysfunction in China. A network pharmacology method was performed systematically, at a molecular level, to analyse the pharmacological mechanism of Yinyanghuo as erectile dysfunction therapy. The network pharmacology method used in this study primarily includes prescreening of the active compounds, prediction of targets, network analysis and gene enrichment analysis. This network analysis proved that 4 targets (AR, NR3C2, PDE5A and BMP2) could be the targets of Yinyanghuo therapy on erectile dysfunction. Besides, gene enrichment analysis predicted that Yinyanghuo might have a role in erectile dysfunction by regulating 10 molecular functions, 8 cellular components, 10 biological processes and 36 possible targets related to 10 signalling pathways. Our study demonstrated the molecular and pharmacological mechanisms of Yinyanghuo against erectile dysfunction with a holistic approach and demonstrated a powerful method for analysing pharmacological mechanisms and rational utilisation of Traditional Chinese Medicine clinically.     

Effect of Wuzi Yanzong Pills on Sertoli cells and blood–testis barrier in heat-stressed rats based on Akt signalling pathway

The blood–testis barrier (BTB) of Sertoli cells (SCs) is an important biological barrier that maintains spermatogenesis and provides a favourable microenvironment for spermatogenesis. However, heat stress can directly damage the BTB structural proteins of testicular SCs, leading to dyszoospermia. Wuzi Yanzong Pills (WYP) is a traditional Chinese medicine formula used to treat male reproductive diseases. However, whether WYP could ameliorate heat stress injury in primary SCs extracted from rat testes and BTB proteins remains unknown. Here, treatment with WYP (low, medium and high dose) increased the SC viability and the proliferation of cell antigen Ki67 significantly. Additionally, it promoted SC maturation, which presented in the form of increased androgen receptors (ARs) and decreased cytokeratin 18 (CK-18) in three WYP dose groups. WYP upregulated BTB proteins such as zonula occludens 1 (ZO-1) and occludin across all WYP groups and decreased phosphorylated Akt (p-Akt) in the middle and high-dose groups; however, ZO-1 and occludin recovery were reduced with the presence of Akt inhibitor in WYP groups. WYP improved SC viability and proliferation, and ameliorated dedifferentiation and BTB-proteins damaged by heat stress via Akt signalling. The findings present theoretical support for the effects of WYP in the management of dyszoospermia and male infertility.     

Development a nomogram to predict fertilisation rate of infertile males with borderline semen by using semen parameters combined with AMH and INHB

The sperm quality of some males is in a critical state, making it hard for clinicians to choose the suitable fertilisation methods. This study aimed to develop an intelligent nomogram for predicting fertilisation rate of infertile males with borderline semen. 160 males underwent in vitro fertilisation (IVF), 58 of whom received rescue ICSI (R-ICSI) due to fertilisation failure (fertilisation rate of IVF ≤30%). A least absolute shrinkage and selection operator (LASSO) regression analysis identified sperm concentration, progressively motile spermatozoa (PMS), seminal plasma anti-Müllerian hormone (spAMH), seminal plasma inhibin (spINHB), serum AMH (serAMH) and serum INHB (serINHB) as significant predictors. The nomogram was plotted by multivariable logistic regression. This nomogram-illustrated model showed good discrimination, calibration and clinical value. The area under the receiver operating characteristic curve (AUC) of the nomogram was 0.762 (p < .001). Calibration curve and Hosmer–Lemeshow test (p = .5261) showed good consistency between the predictions of the nomogram and the actual observations, and decision curve analysis showed that the nomogram was clinically useful. This nomogram may be useful in predicting fertilisation rate, mainly focused on new biomarkers, INHB and AMH. It could assist clinicians and laboratory technicians select appropriate fertilisation methods (IVF or ICSI) for male patients with borderline semen.     

Mechanism of Graft Damage Caused by NTPDase1-activated Macrophages in Acute Antibody-Mediated Rejection

ObjectivesTo investigate the effect and mechanism of macrophage activation and graft damage caused by nucleoside triphosphate diphosphohydrolase 1 (NTPDase1) in acute antibody-mediated rejection (AMR).MethodsAcute AMR was induced in different skin-grafted nude mouse models with wild-type NTPDase1 expression, transgene-enhanced NTPDase1 expression, or NTPDase1 gene knockout. Several methods (eg, real-time fluorescence quantitative polymerase chain reaction, high-performance liquid chromatography [HPLC], immunofluorescence, flow cytometry, and luciferin/luciferase assays) were used to study (at the histologic and molecular levels) the extracellular adenosine diphosphate (ADP) concentration, macrophage proliferation, major histocompatibility complex (MHC) class II antigen expression on the surface of macrophages, B-cell activating factor (BAFF) expression in the peripheral blood serum, and the total number of SmIg-positive B cells during acute AMR. The relative activity of NTPDase1 in B cells and epithelial cells, pathologic changes, and the incidence of positive C4d deposition around the capillaries of skin grafts on the different nude mice were studied.ResultsMacrophages proliferated significantly when acute AMR occurred. The higher the NTPDase1 expression level, the lower the extracellular ADP concentration, the expression of MHC class II antigens on the surface of macrophages, the expression of BAFF in the peripheral blood serum, and the total number of SmIg-positive B cells, indicating negative correlations. The relative activity of NTPDase1 in B cells and epithelial cells of the skin graft was different among the different mice. The higher the NTPDase1 expression level, the lower the degree of pathologic damage to the skin graft.ConclusionsImbalance in extracellular ADP degradation by NTPDase1 may promote macrophage activation, and activated macrophages may be an important cause of graft damage.