弓形虫GRA6可溶性蛋白在大肠埃希菌中的表达及纯化

目的在大肠埃希菌中高效表达及纯化弓形虫GRA6抗原,为制作弓形虫感染基因工程诊断试剂盒奠定基础.方法将重组pGEX-GRA6表达载体转化大肠埃希菌BL21-Codon Plus（DE3）-RP菌株,在异丙基硫代-β-D半乳糖苷（IPTG）诱导下表达.超声破壁后,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析表达产物的表达形式,并对表达产物以硫酸铵沉淀、Sephedax G50脱盐和GST亲和层析柱进行目的蛋白的纯化.通过Western blotting检测其纯化的重组抗原的免疫反应性.结果GRA6以融合蛋白（GST-GRA6）的形式在大肠埃希菌中得到了高效表达.对表达产物可溶性分析表明,表达蛋白在上清液中和包涵体中均有表达.在上清液中表达的可溶性蛋白经纯化后蛋白纯度可达90%以上.Western blotting分析表明纯化蛋白能被弓形虫感染的人血清所识别.结论GRA6在大肠埃希菌中得到了高效表达,重组抗原经纯化后能特异性地被弓形虫感染者血清所识别.     

清热理气冲剂对实验性胆汁反流性胃炎大鼠血浆内皮素和降钙素基因相关肽的影响

目的 探讨清热理气冲剂对实验性胆汁反流性胃炎（BRG）大鼠血浆内皮素（ET）和降钙素基因相关肽（CGRP）的影响。方法 60只大鼠随机分为A组（中药等效剂量组），B组（中药大剂量组），C组（西沙必利等效剂量组），D组（枸橼酸铋钾等效剂量组），E组（病理对照组）和F组（正常对照组）。除F组外，其余各组大鼠应用胆汁灌胃导致胃黏膜损害。2W后分别应用清热理气冲剂等效剂量、大剂量、西沙必利、枸橼酸铋钾处理。实验第4周观察胆汁反流性胃炎胃黏膜病理形态和应用放免法测定血浆ET和CGRP含量。结果 F组可见黏膜下层充血水肿，胃小凹增生，肠上皮化生，炎性细胞浸润等，而A组和B组胃黏膜组织形态显著改善。F组电镜观察到胃黏膜细胞问的紧密连接受损，线粒体肿胀，粗面内质网扩张，微绒毛变短或丢失，A组和B组的超微结构明显的改善。A组和B组血浆ET明显低于E组（P〈0．01）。A组和B组血浆CGRP明显高于E组（P〈0．01）。结论 清热理气冲剂可以减轻胃黏膜损伤，通过影响血浆ET和CGRP相关肽含量而达到治疗BRG目的。     

裸花紫珠颗粒联合替硝唑、头孢呋辛钠治疗慢性盆腔炎患者的疗效评价

目的分析裸花紫珠颗粒联合替硝唑氯化钠注射液、头孢呋辛钠抗菌药物治疗慢性盆腔炎(CPID)患者的临床效果。方法选取2017年1月~2018年12月某院CPID患者126例,随机分为研究组(n=63)与对照组(n=63),对照组予以替硝唑氯化钠注射液、头孢呋辛钠治疗,研究组在对照组基础上加用裸花紫珠颗粒。对比两组疗效、临床症状改善时间(发热、白带异常、下腹痛)及治疗前后血清炎性因子水平[白细胞介素-1β(IL-1β)、单核细胞趋化蛋白-1(MCP-1)]变化。结果研究组总有效率92.06%(58/63)较对照组79.37%(50/63)高(P<0.05);研究组发热、白带异常、下腹痛改善时间较对照组短(P<0.05);治疗后,研究组血清IL-1β、MCP-1水平较对照组低(P<0.05)。结论裸花紫珠颗粒联合替硝唑氯化钠注射液、头孢呋辛钠抗菌药物治疗CPID患者效果显著,可减轻炎症反应,改善患者临床症状。     

Dual Differentiation in Small Cell Carcinoma (Oat Cell Carcinoma) of the Lung

Two cases of small cell “undifferentiated” carcinoma (oat cell carcinoma) of the lung with dual neurosecretory and squamous differentiation are reported. One was treated with various regimens of chemotherapy and radiotherapy; the other was untreated at the time of biopsy. By electron microscopy, the same tumor cells showed membrane-bound, dense-core, neurosecretory-type granules and bundles of tonofibrils denoting squamous differentiation. This dual differentiation underlies the difficulty of ascribing an exclusively neural histogenesis to small cell carcinomas with neurosecretory-type granules. It is possible that entoderm-associated neuroectodermal tumors develop features of autochthonous entodermal tumors as a result of microenvironmental influences. An alternative hypothesis would be that small cell tumors are derived from basal reserve bronchoepithelial cells with pluripotentialities.     

Crystal-Deficient Alveolar Soft Part Sarcoma

Classically, ultrastructural examination of alveolar soft part sarcoma reveals large, dramatic, rhomboid to needlelike crystals with a characteristic substructure. In this study of four cases of alveolar soft part sarcoma, only two exhibited large crystals, which were rare. All four cases, however, exhibited round, electron-dense granules, and in the two cases without large crystals these granules rarely exhibited elongation with the characteristic substructure of alveolar soft part sarcoma that permits definitive diagnosis. Two of these cases had been previously studied at other institutions, where crystals were not identified ultrastructurally and electron microscopy was considered noncontributory. Large crystals, then, may be rare or absent in alveolar soft part sarcoma. Careful search may be necessary to find granules with the characteristic periodic substructure.     

Paclitaxel-induced stress granules increase LINE-1 mRNA stability to promote drug resistance in breast cancer cells

Abnormal expression of long interspersed element-1 (LINE-1) has been implicated in drug resistance, while our previous study showed that chemotherapy drug paclitaxel (PTX) increased LINE-1 level with unknown mechanism. Bioinformatics analysis suggested the regulation of LINE-1 mRNA by drug-induced stress granules (SGs). This study aimed to explore whether and how SGs are involved in drug-induced LINE-1 increase and thereby promotes drug resistance of triple negative breast cancer (TNBC) cells. We demonstrated that SGs increased LINE-1 expression by recruiting and stabilizing LINE-1 mRNA under drug stress, thereby adapting TNBC cells to chemotherapy drugs. Moreover, LINE-1 inhibitor efavirenz (EFV) could inhibit drug-induced SG to destabilize LINE-1. Our study provides the first evidence of the regulation of LINE-1 by SGs that could be an important survival mechanism for cancer cells exposed to chemotherapy drugs. The findings provide a useful clue for developing new chemotherapeutic strategies against TNBCs.     

小儿肺咳颗粒联合孟鲁司特钠治疗小儿肺炎的疗效及对症状改善和睡眠的影响

目的:分析小儿肺咳颗粒联合孟鲁司特钠治疗小儿肺炎的疗效。方法:选取2018年1月至2019年6月广州中医药大学第一附属医院白云医院儿科收治的小儿肺炎患儿50例作为研究对象,随机分为观察组和对照组,每组25例。对照组行常规治疗+孟鲁司特钠,观察组在其基础上联用小儿肺咳颗粒,比较2组疗效。结果:治疗前比较,2组睡眠质量评分比较差异无统计学意义(P>0.05),观察组体温恢复时间、咳嗽咳痰改善和肺部啰音消失时间均短于对照组(P<0.05);治疗后2周、4周2组PSQI得分较治疗前明显下降且观察组PSQI得分低于对照组(P<0.05)。结论:在小儿肺炎治疗中联合小儿肺咳颗粒和孟鲁司特钠可以缩短患者症状恢复时间,提高睡眠质量,值得应用。     

补青颗粒联合石决明水提液对白内障大鼠眼组织细胞中TFAR19和JNK3蛋白表达的影响

目的探讨补青颗粒联合石决明水提液对白内障大鼠眼组织细胞中TF-1细胞凋亡相关基因19(TFAR19)和c-Jun氨基末端激酶(JNK3)蛋白表达的影响。方法选择无特定病原体(SPF)级SD大鼠48只,雌雄各半,根据随机数字表法分为4组:正常组、白内障模型组、阳性对照组和实验组,每组各12只。正常组采用生理盐水滴眼,其余3组均通过亚硒酸钠诱导白内障模型,模型建立后白内障模型组采用生理盐水滴眼,阳性对照组采用莎普爱思滴眼和实验组采用0.25%石决明水提液滴眼+2 g/mL补青颗粒水溶液灌胃,灌胃给药每日2次,滴眼每日1次,每次1滴,持续干预30 d。分别于干预3、10、20、30 d后,蛋白质印迹法检测各组大鼠晶状体组织中TFAR19、JNK3以及pJNK3的表达,检测各组大鼠血清氧化应激指标丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)、总超氧化物歧化酶(T-SOD)含量。结果干预3、10、20、30 d后,阳性治疗组和实验组晶状体组织中TFAR19相对表达量和p-JNK3/JNK3比值均随治疗时间延长而显著降低(P<0.05)。干预20~30 d后,4组大鼠晶状体组织中TFAR19相对表达量为:正常组<实验组<阳性对照组<白内障模型组,且组间两两比较,差异有统计学意义(P<0.05)。干预30 d后,4组大鼠视网膜组织中p-JNK3/JNK3水平比较:正常组<实验组<阳性对照组<白内障模型组,且组间两两比较差异有统计学意义(P<0.05)。各组大鼠血清中MDA水平比较:正常组<实验组<阳性对照组<白内障模型组,GSH-Px和T-SOD则为:白内障模型组<阳性对照组<实验组<正常组,且组间两两比较差异均有统计学意义(P<0.05)。结论补青颗粒联合石决明水提液可通过抑制大鼠晶状体TFAR19蛋白表达和视网膜JNK3磷酸化,并进一步调节氧化应激反应水平,从而对白内障大鼠发挥治疗作用,且在本研究时间范围内其作用效果随时间增加而增强。     

养胃颗粒联合四联疗法治疗Hp阳性慢性胃炎的疗效

目的探讨养胃颗粒联合四联抗菌法治疗Hp阳性慢性胃炎的疗效及其安全性。方法选取2018年12月至2019年12月Hp相关性慢性胃炎患者120例,按照随机数字法分为对照组和观察组,各60例。对照组给予四联疗法,观察组给予养胃颗粒联合四联疗法,比较两组患者治疗有效率、Hp根除率、血液指标变化及不良反应情况。结果观察组临床治疗总有效率、Hp根除率、血清GAS水平、血清PGⅠ水平和PGⅠ/PGⅡ比值均高于对照组,差异有统计学意义(P<0.05)。结论养胃颗粒联合四联抗菌法治疗Hp阳性慢性胃炎的临床效果更优越,能显著提高Hp根除率,并可以减少消化道不良反应和肝损病例。