养阴利咽饮与玉液散吹喉对阴虚肺燥型慢性咽炎患者血清因子的影响

目的:探讨养阴利咽饮与玉液散吹喉对阴虚肺燥型慢性咽炎患者血清炎性因子的影响。方法:将2015年8月至2017年3月山东省淄博市第四人民医院收治的慢性咽炎患者118例作为研究对象,按治疗方法不同分为对照组和观察组,每组59例。对照组用常规西药治疗,观察组用养阴利咽饮与玉液散吹喉治疗,均连续治疗30 d。比较2组临床疗效,观察并比较治疗前后2组患者主要临床症状改善情况,唾液中分泌型免疫球蛋白A(SIgA)、血清炎性因子水平及生命质量变化。结果:治疗后观察组总有效率为91.53%,高于对照组的76.27%(P<0.05)。治疗后2组咽痛、干咳、咽黏膜充血水肿临床症状积分低于治疗前,且观察组低于对照组(P<0.01)。与治疗前比较,治疗后2组患者唾液中SIgA水平均明显升高,血清肿瘤坏死因子-α(TNF-α)及C-反应蛋白(CRP)水平均明显降低,且2组比较差异均有统计学意义(均P<0.01)。治疗后2组患者SF-36各项评分均明显高于治疗前,且观察组高于对照组(P<0.05或P<0.01)。结论:养阴利咽饮与玉液散吹喉联用可降低阴虚肺燥型慢性咽炎患者血清TNF-α、CRP水平,升高唾液中SIgA含量,改善患者临床症状,提高患者生命质量。     

银杏内酯B对肌萎缩侧索硬化细胞模型的保护作用及其机制

目的:探讨银杏内酯B(ginkgolide B,GB)对肌萎缩侧索硬化细胞模型c-Jun氨基末端激酶(JNK)信号通路及细胞凋亡的影响。方法:通过含过表达人突变超氧化物歧化酶1(SOD1)~(G93A)基因(hSOD1~(G93A))和过表达人野生SODWT基因(hSOD1WT)与空质粒的慢病毒感染NSC34细胞,经一定浓度的嘌呤霉素筛选,倒置荧光显微镜下观察慢病毒的转染效率和细胞形态的变化,蛋白免疫印迹法(Western blot)检验感染细胞是否过表达SOD1蛋白,建立hSOD1~(G93A)-NSC34细胞系后给予GB,细胞培养分组为正常组、模型组、不同浓度GB组(25,50,75,100 mg·L-1)组,48 h后噻唑蓝(MTT)比色法检测细胞存活率,筛选出最佳药物浓度,后续实验分组为以正常组,模型组,75 mg?L~(-1) GB组,SP600125组,75 mg?L-1GB+SP600125组,流式细胞术检测各组细胞的凋亡率,蛋白免疫印迹法(Western blot)检测磷酸化(p-)JNK,c-Jun,p-c-Jun,半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)蛋白的表达。结果:与正常的NSC34细胞比较,hSOD1~(G93A)NSC34组细胞胞体变圆,突触减少、变短,而hSOD1WT NSC34组细胞和空质粒组细胞形态学未发生明显变化,与正常组比较,hSOD1~(G93A)NSC34组,hSOD1WTNSC3组细胞内SOD1蛋白水平显著升高(P0. 01),肌萎缩侧索硬化(ALS)细胞模型成功建立。与正常组比较,模型组细胞存活率显著降低(P0. 01);与模型组比较,给予不同浓度GB后,细胞存活率均显著升高(P0. 01),药物质量浓度为75 mg?L-1时细胞存活率显著升高(P0. 01)。后续实验,与正常组比较,模型组凋亡率,p-JNK,p-c-Jun,cleaved Caspase-3蛋白表达量显著升高(P0. 01);与模型组比较,75 mg?L-1GB组,SP600125组,75 mg?L-1GB+SP600125组凋亡率,p-JNK,p-c-Jun,cleaved Caspase-3蛋白表达明显降低(P0. 05,P0. 01)。结论:GB对肌萎缩侧索硬化细胞模型具有抑制细胞凋亡的保护作用,这种保护作用可能是通过JNK信号通路实现的。     

牛奶相关症状评分在婴儿牛奶蛋白过敏早期识别中的评价：多中心调查分析

目的评价牛奶相关症状评分(CoMiSS)在早期识别婴儿牛奶蛋白过敏(CMPA)中的作用。方法 2016年6月至2017年5月对6家儿科门诊疑似CMPA的210例婴儿进行CoMiSS评分,通过开放性食物激发试验(OFC)明确或排除CMPA诊断。同期选取96例健康体检婴儿为对照组。通过ROC曲线评估CoMiSS识别CMPA的灵敏度和特异度,评价其在早期识别婴儿CMPA中的作用。结果 210例疑似CMPA婴儿中,男114例、女96例,中位年龄4.5个月(1～12个月);其中82例确诊CMPA,中位CoMiSS为7.0分(6.0～8.3分);128例非CMPA婴儿的中位CoMiSS为4.0分(3.0～5.0分)。96例对照组中,男42例、女54例,中位年龄4.1个月(1～12个月);中位CoMiSS为 3.0分(2.0～4.0分)。三组间CoMiSS差异有统计学意义(H=152.58,P0.01)。CoMiSS的ROC曲线下面积(AUC)为0.90,最佳诊断界值为5.5,灵敏度为87.8%,特异度为78.1%。结论 CoMiSS评分临床可操作性强,适合儿科推广。建议以CoMiSS≥6分作为早期识别CMPA标准。     

Paraneoplastic opsoclonus myoclonus syndrome associated with inflammatory myofibroblastic tumor in a pediatric patient

Opsoclonus myoclonus syndrome (OMS) is a rare neurological syndrome caused by a paraneoplastic autoimmune process that affects children with neuroblastic tumors. Treatment includes corticosteroids, intravenous gamma globulin (IVIG), rituximab, and other immunosuppressive therapies. Here, we describe a patient diagnosed with OMS associated with a localized inflammatory myofibroblastic tumor. The patient has no evidence of tumor recurrence following surgical resection with 8‐month follow‐up. The neurologic symptoms resolved with corticosteroids and IVIG. This case demonstrates that in children, neoplasms other than neuroblastoma may be associated with this paraneoplastic syndrome, and highlights the importance of evaluating patients with OMS for underlying malignancies.     

Anticonvulsant effect of melatonin through ATP-sensitive channels in mice

Melatonin is a neurohormone secreted principally by the pineal gland. This molecule has various pharmacological properties including improving immune system, prevent cancer, anti-aging, and anti-oxidant effects. The anticonvulsant effects of melatonin have been proved by previous studies. Adenosine triphosphate (ATP)-sensitive potassium (K_ATP) channels are considered as an important target in the seizure modulation. The aim of the present study was to investigate the anticonvulsant effect of melatonin in pentylenetetrazole (PTZ)-induced seizures in mice, focusing on its ability to regulate K_ATP channels. Acute intraperitoneal administration of melatonin (40 and 80 mg/kg) increased clonic seizure threshold induced by intravenous administration of PTZ. Melatonin (40 and 80 mg/kg) increased the latency of clonic seizure and reduced its frequency in mice receiving an intraperitoneal injection of PTZ. Administration of glibenclamide, a K_ATP channels blocker, before intravenous injection of PTZ reduced melatonin anticonvulsant effect. Diazoxide and cromakalim, as K_ATP channels openers, increased antiseizure effect of melatonin in PTZ model of seizures. These findings suggest that the antiseizure effect of melatonin probably is gained through increasing the opening of K_ATP channels.     

Comparing the efficiency of different Escherichia coli strains in producing recombinant capsid protein of porcine circovirus type 2

The present study was aimed at comparing different E. coli strains in expressing the capsid protein of Porcine Circovirus 2 (PCV2). Full length capsid protein could be expressed only in Rosetta-gami 2 (DE3) pLysS strain using pET32b (+) vector. This confirmed that only those strains which possess tRNAs for rare codons can express the full length capsid protein. Purification of full length capsid protein could not be achieved even after several attempts using native and denaturing conditions. Subsequently, an attempt was made for expression of N-terminal truncated capsid protein using the same expression system. Truncated capsid protein was successfully expressed, purified and characterized by western blotting. The truncated capsid protein was also shown to be efficacious in testing serum samples using an optimized indirect ELISA, wherein a diagnostic sensitivity of 88.89% and specificity of 90.82% was obtained as compared to commercially available GreenSpring® porcine circovirus (PCV2) ELISA test kit. Thus, the expressed truncated capsid protein appears to be a promising diagnostic agent for PCV2. The comparative analysis suggests that cluster of arginine residues at N-terminal of capsid protein not only affects its expression in some E. coli strains but also its purification by Ni-NTA chromatography, when expressed as a histidine tagged fusion protein.     

基于p38 MAPK信号通路探讨苍膝通痹胶囊对KOA大鼠关节软骨的保护作用

目的:观察苍膝通痹胶囊治疗膝骨关节炎(KOA)模型大鼠的疗效及相关作用机制。方法:将60只4周龄SPF级健康雄性SD大鼠,随机分为空白组,模型组,二甲基亚砜(DMSO)组,苍膝通痹胶囊组,SB203580组以及苍膝通痹胶囊联合SB203580组,每组10只。除正常组外,其余各组采用改良Hulth法建立KOA模型,造模成功后,苍膝通痹胶囊组每日给予0.25 g·kg~(-1)苍膝通痹胶囊溶液灌胃,SB203580组每日给予0.015 g·kg~(-1)的SB203580溶液灌胃,苍膝通痹胶囊联合SB203580组组每日给予含0.015 g·kg~(-1)SB203580及0.25 g·kg~(-1)苍膝通痹胶囊的混合溶液灌胃,DMSO组给予1%DMSO溶液灌胃,模型组和空白组给予生理盐水灌胃,用药干预4周后处死、取材。苏木素-伊红(HE)染色观察软骨组织形态学改变,酶联免疫吸附测定(ELISA)检测外周血上清液中白细胞介素-1β(IL-1β),肿瘤坏死因子-α(TNF-α)表达水平,实时荧光定量PCR(Real-tine PCR)及蛋白免疫印迹法(Western blot)检测软骨组织中p38 MAPK信号通路中相关因子p38,p-p38,基质金属蛋白酶-13(MMP-13),Ⅱ型胶原蛋白(CollagenⅡ)mRNA及蛋白的表达水平,免疫组化法检测p-p38的定位表达。结果:与正常组比较,模型组关节软骨中p38,p-p38,MMP-13表达水平显著上调(P0.01),CollagenⅡ表达水平显著下调(P0.01),血清IL-1β,TNF-α表达水平显著上调(P0.01);与模型组比较,苍膝通痹胶囊组,SB203580组以及苍膝通痹胶囊联合SB203580组关节软骨中p38,p-p38,MMP-13表达水平显著下调(P0.01),CollagenⅡ表达水平显著上调(P0.01),血清IL-1β,TNF-α表达水平显著下调(P0.01)。结论:苍膝通痹胶囊可有效保护KOA大鼠软骨组织,其机制可能与靶向阻断p38 MAPK信号通路有关。     

Clinicopathological analysis of immunohistochemical expression of CD47 and SIRPα in adult T-cell leukemia/lymphoma

The interaction of CD47 and signal-regulatory protein alpha (SIRPα) induces “don't eat me signal”, leading suppression of phagocytosis. This signal can affect the clinical course of malignant disease. Although CD47 and SIRPα expression are associated with clinicopathological features in several neoplasms, the investigation for adult T-cell leukemia/lymphoma (ATLL) has not been well-documented. This study aimed to declare the association between CD47 and SIRPα expression and clinicopathological features in ATLL. We performed immunostaining on 73 biopsy samples and found that CD47 is primarily expressed in tumor cells, while SIRPα is expressed in non-neoplastic stromal cells. CD47 positive cases showed significantly higher FoxP3 (P = .0232) and lower CCR4 (P = .0214). SIRPα positive cases presented significantly better overall survival than SIRPα negative cases (P = .0132). SIRPα positive cases showed significantly HLA class I (P = .0062), HLA class II (P = .0133), microenvironment PD-L1 (miPD-L1) (P = .0032), and FoxP3 (P = .0229) positivity. In univariate analysis, SIRPα expression was significantly related to prognosis (Hazard ratio [HR] 0.470; 95% confidence interval [CI] 0.253-0.870; P = .0167], although multivariate analysis did not show SIPRα as an independent prognostic factor. The expression of SIRPα on stromal cells reflects activated immune surveillance mechanism in tumor microenvironment and induce good prognosis in ATLL. More detailed studies for gene expression or genomic abnormalities will disclose clinical and biological significance of the CD47 and SIRPα in ATLL.