百令胶囊联合吗替麦考酚酯对过敏性紫癜性肾炎患儿肾功能、体液免疫功能及血清炎性因子的影响

摘要 目的：探讨百令胶囊联合吗替麦考酚酯对过敏性紫癜性肾炎（HSPN）患儿肾功能、体液免疫功能及血清炎性因子的影响。方法：选取2014年7月~2020年2月期间我院收治的80例HSPN患儿,按照随机数字表法分为对照组（n=40,吗替麦考酚酯治疗）和观察组（n=40,百令胶囊联合吗替麦考酚酯治疗）,对比两组疗效、肾功能[24 h尿蛋白定量（24 h Upro）、血肌酐（Scr）、尿素氮（BUN）]、体液免疫功能[免疫球蛋白(Ig)M、IgA、IgG]及血清炎性因子[γ-干扰素（IFN-γ）、白介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）],记录两组治疗期间不良反应情况。结果：与对照组相比,观察组的总有效率更高（P<0.05）。两组不良反应发生率对比未见差异（P>0.05）。治疗3个月后,两组Scr、24 h Upro、BUN、IgM、IgA、IgG、IFN-γ、IL-6、TNF-α均较治疗前下降,且观察组较对照组低（P<0.05）。结论：百令胶囊联合吗替麦考酚酯治疗HSPN患儿疗效值得肯定,可有效改善患儿肾功能、体液免疫功能,减轻患儿炎性反应,且安全性较好。     

Production of target-specific recombinant human polyclonal antibodies in mammalian cells

We describe the expression and consistent production of a first target-specific recombinant human polyclonal antibody. An anti-Rhesus D recombinant polyclonal antibody, Sym001, comprised of 25 unique human IgG1 antibodies, was produced by the novel Sympress expression technology. This strategy is based on site-specific integration of antibody genes in CHO cells, using the FRT/Flp-In recombinase system. This allows integration of the expression construct at the same genomic site in the host cells, thereby reducing genomic position effects. Different bioreactor batches of Sym001 displayed highly consistent manufacturing yield, antibody composition, binding potency, and functional activity. The results demonstrate that diverse recombinant human polyclonal antibody compositions can be reproducibly generated under conditions directly applicable to industrial manufacturing settings and present a first recombinant polyclonal antibody which could be used for treatment of hemolytic disease of the newborn and/or idiopathic thrombocytopenic purpura.     

Escherichia coli-derived von Willebrand factor-A2 domain fluorescence/Förster resonance energy transfer proteins that quantify ADAMTS13 activity

The cleavage of the A2 domain of von Willebrand factor (VWF) by the metalloprotease ADAMTS13 regulates VWF size and platelet thrombosis rates. Reduction or inhibition of this enzyme activity leads to thrombotic thrombocytopenic purpura (TTP). We generated a set of novel molecules called VWF-A2 FRET (fluorescence/Förster resonance energy transfer) proteins, where variants of yellow fluorescent protein (Venus) and cyan fluorescent protein (Cerulean) flank either the entire VWF-A2 domain (175 amino acids) or truncated fragments (141, 113, and 77 amino acids) of this domain. These proteins were expressed in Escherichia coli in soluble form, and they exhibited FRET properties. Results show that the introduction of Venus/Cerulean itself did not alter the ability of VWF-A2 to undergo ADAMTS13-mediated cleavage. The smallest FRET protein, XS-VWF, detected plasma ADAMTS13 activity down to 10% of normal levels. Tests of acquired and inherited TTP could be completed within 30 min. VWF-A2 conformation changed progressively, and not abruptly, on increasing urea concentrations. Although proteins with 77 and 113 VWF-A2 residues were cleaved in the absence of denaturant, 4 M urea was required for the efficient cleavage of larger constructs. Overall, VWF-A2 FRET proteins can be applied both for the rapid diagnosis of plasma ADAMTS13 activity and as a tool to study VWF-A2 conformation dynamics.     

腹型过敏性紫癜患儿急性期与恢复期肠道菌群变化的分析

目的探究腹型过敏性紫癜患儿急性期与恢复期肠道菌群的变化,为制定更针对性的过敏性紫癜治疗措施提供依据。方法选取28例腹型过敏性紫癜患儿粪便标本(急性期19例和恢复期9例),9例非腹型过敏性紫癜患儿粪便标本,一共37例粪便标本。采用Illumina MiSeq平台提取样本DNA进行高通量测序,对测序结果进行生物信息学分析。结果腹型过敏性紫癜患儿急性期肠道菌群α多样性与恢复期、非腹型过敏性紫癜存在显著差异,而β多样性没有显著差异,腹型过敏性紫癜患儿急性期粪便标本中韦荣球菌属的相对丰度显著高于无腹痛腹型过敏性紫癜和非腹型过敏性紫癜患儿,而瘤胃球菌属的相对丰度在腹型过敏性紫癜患儿恢复期粪便标本中最高。结论肠道菌群的变化可能参与腹型过敏性紫癜的发病,其中主要表现在急性期韦荣球菌属丰富度的增加和恢复期瘤胃球菌属丰富度的升高,为微生物制剂的应用提供依据。     

儿童过敏性紫癜的临床特点及其肾损害的相关因素分析

摘要目的：总结儿童过敏性紫癜的临床特点,并分析其肾损害的相关因素。方法：选择2006年2月~2012年8月我院小儿科诊治的过敏性紫癜患儿86例,根据是否伴有肾脏损害分为观察组（34例）和对照组（52例）,分析两组患者的一般临床资料,总结儿童过敏性紫癜的临床特点,并探讨导致儿童过敏性紫癜并发肾损害的相关因素。结果：（1）儿童过敏性紫癜的诱因包括感染、过敏、疫苗接种、寄生虫病史及其它原因,其中感染比例最高,占66．28％;首发症状包括紫癜、腹痛及关节痛的一种或多种,其中紫癜比例最高,占59．30％。（2）观察组患儿中年龄≥8Y、皮疹反复≥4w及血FIB升高的比例分别为73．53％（25／34）、52．94％（18／34）和26．47％（9／34）,均显著高于对照组患儿（P〈0．05）,非条件Logistic多元回归分析结果示皮疹反复≥4W及血FⅢ升高为过敏性紫癜患儿发生肾脏损伤的危险因素。结论：儿童过敏性紫癜的主要诱因为感染,典型临床表现为紫癜,皮疹反复芝4w及血FIB升高为过敏性紫癜患儿发生肾脏损伤的危险因素。     

Relation between 3435C>T multidrug resistance 1 gene polymorphism with high dose methylprednisolone treatment of childhood acute idiopathic thrombocytopenic purpura

The current study was conducted to assess 3435C>T multidrug resistance 1 gene polymorphism and the efficacy of high dose methylprednisolone (HDMP) in childhood acute idiopathic thrombocytopenic purpura patients.

Methods

A total of 31 childhood acute Idiopathic thrombocytopenic purpura patients (17 females, 14 males) between the ages of 2 and 16 years of age were included in the study. High-dose methylprednisolone was given at a dose of 30 mg/kg/day for 3 days and 20 mg/kg/day for 4 days, consecutively and intravenously. Polymerase chain reaction-restriction fragment length polymorphism was used for the detection of C3435T single nucleotide polymorphism. Fragments obtained were 238 bp to T/T genotype, 172 bp and 60 bp fragments to the C/C genotype, and 238 bp, 172 bp and 60 bp to the C/T genotype.

Results

The distribution of CC, CT, and TT genotypes were 19.0%, 61.3%, and 19.4%, respectively. Both allele frequencies of C and T were the same — 50%. There was no significant difference in genotype and allele distribution between the patients with ITP and the control group (χ² = 0.84 p = 0.65, χ² = 0.2 p = 0.63, respectively). There were no significant differences in age, gender, and pre- and post-treatment platelet counts between CC, CT, and TT genotypes of the MDR gene. Response to treatment shows no significant difference between genotype and allele groups.

Conclusion

In our study, there was no difference in the HDMP treatment response between MDR1 gene genotypes. However, it should be noted that this study includes a small group of patients. Our data should therefore be considered preliminary, awaiting further confirmatory studies on an expanded patient base.     

不同剂量丙种球蛋白联合地塞米松治疗儿童特发性血小板减少性紫癜的疗效观察

目的:探讨不同剂量丙种球蛋白联合地塞米松治疗儿童特发性血小板减少性紫癜(ITP)的临床疗效。方法:选择2010年7月至2015年7月间我院收治的特发性血小板减少性紫癜患儿78例,按照随机数字表法将其分为观察组38例及对照组40例。观察组采用200 mg/(kg·d)丙种球蛋白联合地塞米松进行治疗,对照组采用400 mg/(kg·d)丙种球蛋白联合地塞米松进行治疗。对两组患者的临床疗效、治疗前后血小板计数、血小板上升时间、血小板恢复正常时间、出血停止时间、住院时间、不良反应及医疗费用进行观察。结果:两组患儿血小板计数治疗前无显著差异(P0.05),治疗后两组血小板计数均呈逐渐升高趋势(P0.05),但组间比较无统计学差异(P0.05)。两组患儿血小板上升时间、血小板恢复正常时间、出血停止时间及住院时间均接近,无统计学差异(P0.05)。观察组治疗总有效率为94.74%,对照组为95.00%,两组无显著差异(P0.05)。观察组不良反应发生率为8.17%,对照组为12.50%,两者相比无统计学差异(P0.05)。观察组患儿平均医疗费用为(1.25±0.34)万元,对照组患儿平均医疗费用为(2.31±0.40)万元,观察组明显低于对照组,两者差异有统计学意义(P0.05)。结论:不同剂量丙种球蛋白联合地塞米松治疗ITP具有类似的临床疗效及安全性,但小剂量丙种球蛋白组治疗费用明显降低,因此值得临床推广应用。     

Manufacture of recombinant polyclonal antibodies

Polyclonal antibody therapy in the form of hyper-immune serum has for more than a century been used for treatment of many infectious diseases. However, with the emergence of first antibiotics and later recombinant monoclonal antibody therapy, the use of hyper-immune serum has declined. The main reason for this is that methods for consistent manufacturing of safe hyper immune immunoglobulin products have been lacking. In contrast, manufacturing processes of recombinant monoclonal antibodies follow a well established schedule and it appears obvious to use similar methods to produce recombinant polyclonal products. However, the methods for monoclonal antibody manufacturing are, for several reasons, not directly applicable to generation and manufacture of polyclonal recombinant antibodies. A new production strategy based on recombinant mammalian producer cells has recently been developed to support consistent generation of recombinant polyclonal antibodies for therapeutic use. This review describes aspects of this novel technology with emphasis on the generation, production and characterization procedures employed, and provides comparison with alternative polyclonal and monoclonal antibody manufacturing strategies.     

Immune response to CagA protein is associated with improved platelet count after Helicobacter pylori eradication in patients with idiopathic thrombocytopenic purpura

BACKGROUND: Improvement in platelet counts has been reported after eradication of Helicobacter pylori in patients with idiopathic thrombocytopenic purpura (ITP). We examined the levels of serum markers of gastritis and anti-CagA (cytotoxin-associated gene A) IgG antibody in patients with ITP to investigate whether these factors are associated with the platelet response after H. pylori eradication therapy. MATERIALS AND METHODS: One hundred and sixteen consecutive patients with ITP were assessed for H. pylori infection by (13)C-urea breath test and serum H. pylori antibody test. Patients with H. pylori infection received eradication therapy. Before and after eradication therapy, we evaluated serum levels of gastrin, pepsinogen (PG)-I, and PG-II and the anti-CagA IgG antibody titer. RESULTS: H. pylori infection was found in 67 (58%) of the 116 patients with ITP. Fifty-two infected patients received eradication therapy, which was successful in 44 patients (85%). Twenty-seven patients (62%) showed an increased platelet count and were identified as responders. The duration of ITP was shorter in responders than in nonresponders (p = .017). There was no difference of the levels of gastrin and PGs between responders and nonresponders. Before eradication therapy, the serum anti-CagA antibody titer did not differ significantly between responders and nonresponders. However, reduction in the anti-CagA antibody titer after eradication therapy was significantly greater in responders than in nonresponders (p = .013). CONCLUSIONS: H. pylori eradication therapy improves the platelet count in H. pylori-positive patients with ITP of short duration. Immune response of hosts to CagA protein of H. pylori may play a role in the pathogenesis of ITP.