Cloning and characterization of a novel hepatitis B virus core binding protein C12

AIM: To elucidate the biological function of HBV core antigen (HBcAg) on pathogenesis of hepatitis B, a novel gene C12 coding for protein with unknown function interacting with HBcAg in hepatocytes was identified and characterized. METHODS: HBcAg bait plasmid pGBKT7-HBcAg was constructed and transformed into yeast AH109, then the transformed yeast was mated with yeast Y187 containing liver complementary DNA (cDNA) library plasmid in 2×YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing X-α-gal for screening twice. After extracting and sequencing of plasmid from blue colonies, we isolated a cDNA clone encoding a novel protein designated as C12 that directly interacted with HBcAg. The interaction between HBcAg and C12 was verified again by re-mating. PEGFP-N1-C12 fluorescent protein fusion gene was transfected in 293 and L02 cell, and observed by fluorescent microscope. MTT reduction assay was used to study the action of C12 protein effect on metabolism of mammal cell. Yeast two-hybrid and cDNA microarray were performed to search binding protein and differential expression genes regulated by C12 protein. RESULTS: C12 gene was screened and identified by yeast two-hybrid system 3. The interaction between HBcAg and the novel protein coded by the new gene C12 was further confirmed by re-mating. After 48 h, fluorescence of fusion protein could be observed steadily in the 293 and L02 cell plasma. Under MTT assay, we found that the expression of C12 did not influence the growth of liver cells. Seventeen differential expression genes in HepG2 cells transfected with C12 protein expression plasmid by cDNA microarray, of which 16 genes were upregulated and 1 gene was downregulated by C12 protein. Twenty-one colonies containing 16 different genes coding for C12 protein binding proteins were isolated by yeast two-hybrid, there were 2 new genes with unknown function. CONCLUSION: The novel protein C12 is located in cell plasma, and its overexpression has no significant effect on the metabolism of liver cell. It interacts with many proteins in hepatocytes and may be involved in many processes of gene expression.     

芍药苷对心脏钾通道电生理功能的影响

目的研究芍药苷对内向整流钾电流(IK1)、瞬时外向钾电流(Ito)以及延迟整流钾电流(IKs和IKr)的作用。方法用全细胞膜片钳技术记录大鼠心室肌细胞的Ito和IK1电流。而IKs和IKr电流在转染相应质粒的HEK293细胞上记录。对比芍药苷使用前后的电流图,观察芍药苷对各种离子通道电流的影响。结果在-100mV测试电压下,100μmol/L的芍药苷能使IK1峰值密度从(-25.26±8.21)pA/pF降至(-17.65±6.52)pA/pF,平均抑制率为30.13%(n=6,P<0.05),但对其反转电位以及内向整流特性无影响。此外,100μmol/L芍药苷对Ito、IKs和IKr电流无明显作用。结论芍药苷对IK1电流具有明显的抑制作用,而对Ito、IKs及IKr无明显作用。     

Wy14643对缺氧复氧损伤原代大鼠肝细胞的保护作用及机制

目的探讨过氧化物酶体增殖物激活受体α（PPARα）选择性激动剂Wy14643对原代大鼠肝细胞缺氧复氧损伤的保护作用及机制。方法采用两步灌流法分离大鼠肝细胞并进行原代培养,将培养细胞随机分成6组：C组（正常组）、H/R组（缺氧复氧组）、D组（对照组）、Ⅰ组、Ⅱ组、Ⅲ组。Ⅰ组、Ⅱ组、Ⅲ组均在缺氧前2 h加入Wy14643（终浓度分别是10、30、100μmol/L）再行复氧处理。将肝细胞缺氧4 h、复氧10 h诱导细胞损伤,然后分别收集细胞培养上清液和肝细胞,测定上清液中谷丙转氨酶（ALT）、天门冬氨酸转氨酶（AST）活性和细胞内谷胱甘肽（GSH）、丙二醛（MDA）含量及超氧化物歧化酶（SOD）活性,MTT法测定肝细胞的存活率,电子显微镜观察肝细胞损伤情况。结果与C组比较,H/R组ALT、AST、MDA增高（P〈0.01）,SOD、GSH下降（P〈0.01）,细胞存活率降低（P〈0.01）。Wy14643处理后,肝细胞上清液ALT、AST活性降低,并且呈剂量依赖性（P〈0.01）;肝细胞内SOD、GSH活性增强（P〈0.05）,MDA含量降低（P〈0.05）;细胞存活率升高（P〈0.05）。电子显微镜显示H/R组肝细胞比C组损伤明显,Wy14643处理后损伤逐渐减轻。结论 Wy14643对肝细胞缺氧复氧损伤有保护作用,可能与提高肝细胞抗氧化能力有关。     

肾毒性物质对甲酚抑制内皮祖细胞增殖和eNOS磷酸化

目的:观察对甲酚对人外周血晚期内皮祖细胞(EPCs)的体外增殖和内皮一氧化氮合酶(eNOS)活性的影响.方法:用密度梯度离心法分离健康成人外周血中的单个核细胞,在含有血管内皮生长因子等的培养基中培养.通过形态学、免疫荧光、流式细胞分析鉴定细胞,在贴壁细胞中加入不同浓度对甲酚,用细胞计数和集落生成实验法评价对甲酚对EPC...     

骨蚀宁对激素性股骨头坏死家兔局部rBF、TNF及显微结构的影响

目的：探讨骨蚀宁治疗激素性股骨头坏死的作用机制.方法：选择马血清与甲基强的松龙造成兔激素性股骨头坏死模型,设立对照组、模型组、仙灵骨葆组、骨蚀宁组,测定其血中局部组织血流量（rBF）、肿瘤坏死因子（TNF）,同时取骨在光镜下进行常规病理检查观察.结果：与模型组比较,仙灵骨葆组与骨蚀宁组rBF明显升高（P＜0.01）,但两组之间无明显差异（P＞0.05）.与模型组比较,仙灵骨葆组与模型组TNF含量差别不大（P＞0.05）,而骨蚀宁组升高明显,差异有显著性（P＜0.05）.与仙灵骨葆组比较,骨蚀宁组TNF明显降低,差异有显著性（P＜0.05）.与模型组比较,仙灵骨葆组和骨蚀宁组空骨陷窝百分比均有减少,差异有显著性（P＜0.01）.但均未达正常,组间比较无显著性差异（P＞0.05）.骨蚀宁组股骨头骨小梁和骨细胞的损害明显减轻.结论：骨蚀宁可通过改善股骨头局部组织血流量、抗炎性反应以及保护和促进骨细胞再生达到治疗激素性股骨头坏死的作用.     

结核病的营养治疗

结核病是严重威胁全球健康的疾病之一,其发病与否与营养水平密切相关。结核病多发于贫困地区,人口总体营养水平低下;结核病患者普遍伴有营养不良,不利于结核病的治疗。通过营养治疗,提高患者的营养水平,增强免疫力,优化肠道微生态,提高结核病的治疗效果。本文探讨了营养治疗对结核病治疗的支持作用。     

复发性口腔溃疡患者口腔幽门螺杆菌的检测

目的 探讨复发性口腔溃疡(RAU)的发病与口腔幽门螺杆菌(Hp)感染及消化道疾病之间的关联.方法 以RAU患者唾液为模板,设计引物,用PCR,检测82例RAU患者口腔中Hp阳性率情况.结果 82例RAU患者和74例健康人群,Hp阳性率分别为43.9%和16.2%(P＜0.001);82例RAU患者中,伴有胃肠道疾病的22例,占26.82%;74例健康人群中,伴有胃肠道疾病的7例,占10.45%.比较RAU与非RAU患者中,胃肠道疾病的发病率有显著差异(P＜0.01).结论 Hp感染与RAU的发病之间有某种潜在的关联.RAU患者较健康人群胃肠道疾病患病率高.     

A multicenter retrospective study of transurethral prostate split for benign prostate hyperplasia

BackgroundTransurethral split of the prostate (TUSP) is effective in treating benign prostatic hyperplasia (BPH). However, there is still a lack of research focusing on the optimal target population for TUSP. This study aimed to compare the efficacy of TUSP in patients with different prostate volumes or ages.MethodsThe study was a multicenter retrospective study. The outcomes of TUSP in BPH patients with different prostate volumes or different ages were compared. A total of 439 patients were included in the study. Patients were divided into two groups according to prostate volume, with a cut-off value of 50 mL. Similarly, the cut-off value for the age groups was 70 years. Baseline patient characteristics and perioperative outcomes were recorded. Follow-up was performed at 1, 6, and 12 months after surgery.ResultsThe mean age of the patients was 73.4 years, and the mean prostate volume was 51.2 mL. At 12-month follow-up after TUSP treatment, the patients’ International Prostate Symptom Scores (IPSS), quality of life (QoL) scores, and postvoid residual (PVR) volumes decreased significantly, while peak urinary flow rate (Qmax) increased significantly. Intraoperative hemoglobin (Hb) reduction was significantly lower in the small volume group than in the large volume group. The incidence of postoperative urinary urgency and transient incontinence was lower in the small volume group. IPSS score, PVR, and Qmax in the small volume group showed more remarkable changes at several time points compared to the preoperative period. Postoperative pain scores were higher in the small volume group than in the large volume group. There were no differences between the two groups in terms of long-term complications. The younger group showed greater variation in PVR and Qmax at some time points but less variation in QoL than the older group.ConclusionsTUSP is overall safe and effective in treating BPH. This study showed differences in the outcomes of TUSP in treating different prostate volumes or ages of BPH patients. The optimal surgical approach for BPH patients might be selected clinically based on a combination of prostate volume or patient age.