胞膜(Ecto)-6A8α-甘露糖苷酶

68Aα-甘露糖苷酶是本实验室克隆的cDNA(GenBank的登记号为AF044414)编码的一个新的人α-甘露糖苷酶。该cDNA长3300bp,其开放阅读框架编码1062个氨基酸的蛋白质。蛋白质的1028～1048aa为穿膜区。6A8α-甘露糖苷酶属Ⅰ型穿膜蛋白。6A8基因定位于第15号染色体q11～13区。一般认为,糖基化发生于内质网和高尔基体,     

大量腹水伴多系统受累的TAFRO综合征一例并文献复习

目的 探讨大量腹水伴多系统受累TAFRO综合征患者的识别及临床表现。方法 选取2015年11月航天中心医院出现大量腹水并多系统损伤的患者1例,回顾性分析其入院资料及临床表现、实验室检查结果,并复习相关文献。结果 本例患者女性,39岁,急性起病,发热后出现大量腹水,伴贫血、PLT减少,SCr升高伴少尿。查体：皮肤巩膜黄染,腹膨隆,无压痛及反跳痛,移动性浊音阳性,肠鸣音弱,右下腹留置腹腔引流管,引流液呈血性,双下肢水肿。实验室结果显示,WBC 8.45×10⁹/L,RBC 1.73×10¹²/L,Hb 53 g/L,PLT 11×10⁹/L,BUN 51.6 mmol/L,SCr 232.9μmol/L,免疫球蛋白G 1 730 mg/dl(正常范围：751～1 560 mg/dl),乳酸脱氢酶569.0 IU/L。诊断考虑TAFRO综合征。给予肾脏替代及支持治疗,配合免疫抑制剂,患者临床症状好转。结论 TAFRO综合征与一些自身免疫性疾病背景相同,临床表现为多系统受累,起病急,发病罕见,鉴别比较困难,预后凶险。患者需紧急医...     

人干燥综合征B抗原重组体的构建及融合表达

目的克隆人干燥综合征B抗原基因(human sjogren’s syndrome antigen B,SSB)并进行原核表达,为使用重组抗原用于自身抗体的临床检测奠定基础。方法根据GenBank中检索到的人SSB cDNA序列,在5′非编码区和3′非编码区设计特异性引物,提取人源HeLa细胞总RNA作为模板,逆转录RT-PCR扩增人干燥综合征B抗原cDNA。PCR产物纯化后连接至载体PET-30a,导入大肠埃希菌DH5α,构建重组质粒PET-30a-SSB。对重组质粒进行酶切鉴定,选择阳性克隆测序。重组质粒导入大肠埃希菌BL21,阳性克隆经鉴定后在IPTG诱导下表达。结果RT-PCR扩增产物为1245bp。重组质粒PET-30a-SSB经EcoR Ⅰ和HindⅢ双酶切证实含目的基因片段。序列分析提示与GenBank中检索到的一致。SDS-PAGE和Western印迹结果显示融合蛋白相对分子量为73ku,具有天然SSB抗原活性。结论成功克隆SSB基因并表达融合蛋白。     

Aristolochic acid I-induced apoptosis in LLC-PK1 cells and amelioration of t he apoptotic damage by calcium antagonist

Objective To examine the effect of different concentrations of aristolochic acid I (AAI) in inducing apoptosis of cultured porcine renal cell line LLC-PK1 and to investigate the relationship between intracellular free calcium concentration (［Ca(++) ］i) and LLC-PK1 apoptosis induced by AAI and the influence of a calcium antagonist, lacidipine on apoptosis and ［Ca(++) ］i. Methods LLC-PK1 cells were treated in different groups: a.the normal group without treatment; b.the group with AAI alone (0.01 g·L(-1) , 0.02 g·L(-1) , 0.04 g·L(-1) , 0.08 g·L(-1) ); c.the group with lacidipine alone (10 ng·L(-1) , 10(2) ng·L(-1) ,) 10[3 ng·L(-1) ]; d.the group with AAI (0.04 g·L(-1) ) plus lacidipine (10 ng·L(-1) , 10(2) ng·L(-1) , 10(3) ng·L(-1) ).Light microscopy, agarose gel electrophoresis, Annexin-V-Flous apoptosis detection kit and flow cytometry using propidium iodide staining to identify or quantify the apoptosis of LLC-PK1 cells.Mean ［Ca++］i was measured by laser confocus microscopy using Fluo-3/AM staining. Results A series of morphologic changes that were characteristic of apoptosis, Annexin-V-Flous staining positive apoptotic cells and 'DNA ladder' were identified in AAI　(0.02 g·L(-1) -0.08 g·L(-1) ) treated LLC-PK1 cells.Quantitative analysis of apoptotic cells showed that the percentage of apoptotic cells in AAI (0.02 g·L(-1) , 0.04 g·L(-1) or 0.08 g·L[-1]) group was significantly higher than that in normal group (5.3%, 48.5%, 78.7% vs 2.6%, P<0.001).Mean ［Ca++］i was significantly higher in cells treated with AAI (0.04 g·L(-1) ) than that in normal cells (58.01±18.89 vs 22.66±4.78, P<0.001).In group treated with AAI plus lacidipine (102 ng·L(-1) , 103 ng·L(-1) ), mean ［Ca++］i was significantly lower than that treated with AAI alone (35.47±12.85, 28.55±10.16 vs 58.01±18.89, P<0.001).And the percentage of apoptotic cells in group treated with AAI plus lacidipine ](10(2) ng·L(-1) , 10(3) ng·L(-1) ) was also significantly lower than that treated with AAI alone (19.0%, 27.8% vs 34.7%,P<0.001). Conclusions High concentrations of AAI may induce apoptosis of LLC-PK1 cells.The mean ［Ca++］i in AAI-treated LLC-PK1 cells was increased significantly, sugguesting that the increase of ［Ca++］i may be related to apoptosis in LLC-PK1 cells.Lacidipine may decrease the raised mean ［Ca++］i levels caused by AAI and the percentage of apoptotic cells, and lacidipine may ameliorate AAI-induced apoptotic damage by inhibiting the increase of ［Ca++］i in LLC- PK1 cells.     

维持性血液透析患者透析通路相关缺血综合征发病率及相关因素

目的 调查维持性血液透析患者透析通路相关缺血综合征及窃血现象发病率,对缺血综合征发生的危险因素进行分析.方法 采用横断面研究,选取2012年1至3月在北京市海淀医院以头静脉-桡动脉端侧吻合自体动静脉内瘘为通路行维持性血液透析者71例,询问并检查患者是否存在内瘘侧肢端发凉、发绀等缺血症状;应用彩色多普勒超声观察动静脉内瘘吻合口桡动脉近心端、远心端是否存在反向血流,测量内瘘吻合口、桡动脉、肱动脉内径及血流量;记录入组患者性别、高血压、糖尿病、吸烟和动脉硬化等,对上述指标进行量化后行logistic回归分析.结果 透析通路相关缺血综合征发病率为19.7％（14/71）.彩色多普勒超声显示,桡动脉远端血流方向及频谱方向均为反向者42例（59.2％）.多元回归分析显示,内瘘口径大小、肱动脉血流量、女性、糖尿病、动脉硬化不是缺血综合征的危险因素（均P＞0.05）.结论 动静脉内瘘后,缺血综合征发生率不低,目前尚不能通过控制内瘘口径大小等方法预防其发生.     

1例缺血性卒中合并高血压房颤患者的循证治疗

目的介绍1例缺血性卒中合并高血压房颤患者的循证治疗体会。方法按PICO原则，针对患者的临床问题，计算机检索Cochrane图书馆（2006年第4期）、SUMsearch（1980．1～2006．12）、PubMed（1980．1～2006．12）中有关抗凝剂预防及治疗缺血性脑卒中合并心房纤颤，缺血性卒中降压治疗的循证临床指南、RCT、系统评价和Meta分析，并评价其质量，判断研究结论的真实性，重要性和适用性，获取最佳证据用于临床治疗。结果共检索到4篇指南，3篇系统评价和6篇RCT。分析结果表明：低剂量抗凝治疗对患者安全有效，卒中急性期血压不宜快速降低，根据检索到的证据及患者的具体病情和意愿，该患者抗凝治疗使用小剂量的华法林口服，INR调整为2．0左右，待患者进入恢复期屙再口服福辛普利加吲达帕胺控制血压。经2周低剂量抗凝及辅助康复治疗后患者症状缓解，经过一个月的治疗随访，证实所选方案适合患者。结论对于急性缺血性卒中合并房颤患者，抗凝治疗仍是一线选择，且华法林疗效优于其他药物，针对老年患者应个体化调整INR比值，必要时维持较小值亦可获得最大效益同时减少出血风险，在不能耐受抗凝治疗时可选用阿司匹林治疗。卒中急性期的血压控制不宜太过积极，恢复期可选用ACE抑制剂加小剂量利尿剂控制血压。     

初发系统性红斑狼疮患者调节性T细胞的变化

目的 探讨初发系统性红斑狼疮(SLE)患者CD4 T细胞中FOXP3和CD25的表达及其临床意义.方法 应用流式细胞仪检测CD4 FOXP3 、CD4 CD25 、CD4 CD25high T细胞表达,同时检测CD4 FOXP3 T细胞中CD25及CD4 CD25highT细胞中FOXP3的表达.结果 活动组SLE患者CD4 CD25 T细胞显著低于对照组和低活动组(P＜0.05).活动组患者CD4 CD25highT细胞明显低于低活动组患者(P＜0.01).初发SLE患者CD4 FOXP3 T细胞明显高于对照组(P＜0.01).活动组SLE患者CD4 FOXP3 T细胞中CD25表达显著低于对照组和低活动组(P＜0.05).同时活动组SLE患者CD4 CD25highT细胞中FOXP3表达的平均荧光强度较对照组显著降低(P＜0.05).活动组患者治疗后CD4 CD25highT细胞显著增高(P＜0.05).结论 初发活动期SLE患者CD4 T细胞中CD25high和FOXP3表达不一致可能在SLE患者免疫失衡中发挥作用.     

E3泛素连接酶RNF121对脂多糖诱导的巨噬细胞表达促炎性细胞因子的影响

目的探讨E3泛素连接酶RNF121对脂多糖(LPS)诱导的小鼠腹腔巨噬细胞表达促炎性细胞因子的调控作用。方法 LPS刺激小鼠腹腔巨噬细胞,Western blot检测RNF121的表达。用RNA干扰(si-RNF121)降低RNF121的表达,小鼠腹腔巨噬细胞经LPS刺激后,实时荧光定量PCR法检测TNF-α和IL-6的mRNA水平,ELISA检测细胞培养上清中TNF-α和IL-6的浓度,Western blot检测小鼠腹腔巨噬细胞中P65及磷酸化P65(p-P65)的表达水平,利用双荧光素酶报告基因法检测NF-κB的活性。结果 LPS诱导小鼠腹腔巨噬细胞后,RNF121的蛋白水平显著降低(P0.05),蛋白酶抑制剂MG132能够显著增加RNF121的蛋白水平。si-RNF121降低RNF121的表达后,给予LPS刺激小鼠腹腔巨噬细胞,IL-6和TNF-α的表达水平均显著下降(P0.05),转录因子P65的磷酸化(p-P65)水平及NF-κB的活性均显著降低(P0.05)。结论 E3泛素连接酶RNF121通过调控NF-κB的活性从而影响小鼠巨噬细胞促炎性细胞因子TNF-α及IL-6的表达。     

甲泼尼龙联合环磷酰胺对博来霉素诱导的肺纤维化大鼠模型炎症反应与免疫细胞活性的影响CSCD

Objective To observe the effect of methylprednisolone (MP) combined with cyclo‑ phosphamide (CTX) on inflammation and immune cell activity in bleomycin (BLM)‑induced pulmonary fibrosis rat model. Methods Forty healthy 6 to 8‑week‑old SD rats were randomly divided into blank control, BLM model, BLM+MP, and BLM+MP+CTX groups, with 10 rats in each group. The rat model of pulmonary fibrosis was prepared by intratracheal infusion of BLM (5 mg/kg, only once). From the 7th day of modeling, MP (3 mg/kg) was injected in rats in the BLM+MP group and MP (3 mg/kg)+CTX (8 mg/kg) was injected via tail vein in rats in the BLM+MP+CTX group, once daily for 21 days. The degree of lung inflammation and fibrosis in rats was detected using HE and Masson staining methods. The numbers of granulocytes and neutrophils in bronchoalveolar lavage fluid (BALF) and blood T cell subsets in rats were detected using flow cytometry. Results On the 7th day of modeling, the external morphology, HE and Masson staining results of rat lung tissue showed that BLM‑induced pulmonary fibrosis model was successfully prepared. On the 28th day of modeling, the lung tissue structure of the BLM group was disordered with obvious collagen deposition, the number of granulocytes and neutrophils in BALF increased significantly, the propor‑ tion of blood T cells, CD4+ T cells, and regulatory T cells (Tregs) decreased, the proportion of CD8+ T cells, and the CD4+/CD8+ T cells ratio decreased significantly (all P<0.05). Compared with the BLM group, the degree of pulmonary fibrosis in the BLM+MP+CTX group was improved significantly, the number of granulo‑ cytes and neutrophils in BALF decreased significantly, the proportion of blood T cells, CD4+ T cells and Tregs cells increased significantly, the proportion of CD8+ T cells decreased, and the ratio of CD4+/CD8+ T cells increased significantly (all P<0.05). The improvement effect in rats of BLM+MP+CTX group was better than that of BLM+MP group, and the difference was statistically significant (P<0.05). Conclusion MP com‑ bined with CTX can reduce the degree of inflammatory reaction in rats with pulmonary fibrosis and improve T cell immune activity. © 2023 Chinese Medical Association. All rights reserved.