Synthesis and Characterization of a Full-Length Infectious cDNA Clone of Tomato Mottle Mosaic Virus

Tomato mottle mosaic virus (ToMMV) is a noteworthy virus which belongs to the Virgaviridae family and causes serious economic losses in tomato. Here, we isolated and cloned the full-length genome of a ToMMV Chinese isolate (ToMMV-LN) from a naturally infected tomato (Solanum lycopersicum L.). Sequence analysis showed that ToMMV-LN contains 6399 nucleotides (nts) and is most closely related to a ToMMV Mexican isolate with a sequence identity of 99.48%. Next, an infectious cDNA clone of ToMMV was constructed by a homologous recombination approach. Both the model host N. benthamiana and the natural hosts tomato and pepper developed severe symptoms upon agroinfiltration with pToMMV, which had a strong infectivity. Electron micrographs indicated that a large number of rigid rod-shaped ToMMV virions were observed from the agroinfiltrated N. benthamiana leaves. Finally, our results also confirmed that tomato plants inoculated with pToMMV led to a high infection rate of 100% in 4–5 weeks post-infiltration (wpi), while pepper plants inoculated with pToMMV led to an infection rate of 40–47% in 4–5 wpi. This is the first report of the development of a full-length infectious cDNA clone of ToMMV. We believe that this infectious clone will enable further studies of ToMMV genes function, pathogenicity and virus–host interaction.     

NbTMP14 Is Involved in Tomato Spotted Wilt Virus Infection and Symptom Development by Interaction with the Viral NSm Protein

Tomato spotted wilt virus (TSWV) is one of the most destructive plant viruses, causing severe losses in many important crops worldwide. The non-structural protein NSm of TSWV is a viral movement protein that induces viral symptoms. However, the molecular mechanisms by which NSm contributes to symptom development are unclear. Here, we present evidence that NSm directly interacts with Nicotiana benthamiana chloroplast thylakoid membrane protein TMP14 (NbTMP14) by yeast two-hybrid and bimolecular fluorescence complementation (BiFC) assays. The interaction between NSm and NbTMP14 led to the translocation of the NbTMP14 protein from the chloroplast to the cytoplasm in TSWV-infected plants, and overexpressing NSm decreased NbTMP14 mRNA accumulation. In addition, abnormal chloroplasts and starch accumulation were observed in TSWV-infected plants. Silencing of NbTMP14 by TRV VIGS also showed similar results to those of TSWV-infected plants. Overexpressing NbTMP14 in transgenic N. benthamiana plants impeded TSWV infection, and silencing NbTMP14 in N. benthamiana plants increased disease symptom severity and virus accumulation. To our knowledge, this is the first report showing that the plant chloroplast TMP14 protein is involved in viral infection. Knowledge of the interaction between NSm and NbTMP14 advances our understanding of the molecular mechanisms underlying TSWV symptom development and infection.     

卡介苗HSP70基因转染对小鼠淋巴细胞白血病细胞免疫原性的影响

目的探讨卡介苗热休克蛋白70(BCG HSP70)基因转染对小鼠淋巴细胞白血病细胞(L1210)致瘤性及免疫原性的影响。方法利用脂质体2000将BCG HSP70基因转入小鼠淋巴细胞白血病细胞系L1210表面,获得高表达HSP70分子的L1210-HSP70细胞,作为肿瘤疫苗分别对裸鼠及同系小鼠致瘤性、瘤苗对荷瘤小鼠的免疫治疗作用以及瘤苗对L1210细胞攻击的免疫保护作用进行研究。结果 BCG HSP70成功表达在转染后的L1210细胞表面,并具有与野生型细胞同样的致瘤性。同系小鼠DBA/2皮下接种L1210-HSP70细胞后肿瘤生长缓慢或不成瘤;相对于L1210组和L1210-neo组,生存期明显延长,小鼠脾淋巴细胞中针对L1210细胞的特异性T细胞数量明显增加,差异有统计学意义(P0.05);瘤体病理切片示彻底凝固性坏死,周边可见淋巴样细胞,免疫组化检测示大量CD8+T淋巴细胞浸润。L1210-HSP70细胞瘤苗具有对荷瘤小鼠的免疫治疗作用,以及抗肿瘤攻击的免疫保护作用,均表现为肿瘤出瘤时间明显延长,生长受到显著抑制,瘤体平均直径明显减小;荷瘤小鼠生存时间明显长于L1210组、L1210-neo组和PBS组,差异有统计学意义(P0.05)。结论BCG HSP70基因转染能有效增强肿瘤细胞的免疫原性,激活特异性T细胞,降低肿瘤细胞的致瘤性;并具有提高宿主抗瘤免疫作用。     

An interaction between carotene-15,15'-monooxygenase expression and consumption of a tomato or lycopene-containing diet impacts serum and testicular testosterone

Lycopene, the red pigment of tomatoes, is hypothesized to reduce prostate cancer risk, a disease strongly dependent upon testosterone. In this study, mice lacking the expression of carotene-15,15'-monooxygenase (CMO-I(-/-) ) or wild-type mice were fed either a 10% tomato powder (TP), lycopene-containing (248 nmol/g diet) or their respective control diets for 4 days, after which serum testosterone was measured. A significant diet × genotype interaction (p = 0.02) suggests that the TP reduces serum testosterone concentrations in CMO-I(-/-) mice but not in wild-type mice. Similarly, testicular testosterone was lowered in TP-fed CMO-I(-/-) mice (p = 0.01), suggesting that testosterone synthesis may be inhibited in this group. A similar pattern was also observed for lycopene fed mice. Interestingly, the CMO-I(-/-) mice showed a greater expression of the gene encoding the CMO-II enzyme responsible for eccentric oxidative carotenoid cleavage in the testes. Therefore, we hypothesize that serum testosterone is reduced by lycopene metabolic products of oxidative cleavage by CMO-II in the testes. Overall, these findings suggest that genetic polymorphisms impacting CMO-I expression and its interaction with CMO-II, coupled with variations in dietary lycopene, may modulate testosterone synthesis and serum concentrations. Furthermore, carefully controlled studies with tomato products and lycopene in genetically defined murine models may elucidate important diet × genetic interactions that may impact prostate cancer risk.     

Specificities of Ophthalmic Tumors: Usefulness of A National Network<br/>Spécificités des Tumeurs de la Sphère Ophtalmique: Utilité d’un Réseau National

We describe the most frequent malignant intraocular tumors, conjunctival tumors and some lids and orbital tumors. Primary intraocular malignant tumors are retinoblastoma in children and uveal melanoma in adults. For uveal melanoma, the liver is the most frequent site of metastasis and this is why it is justified to prescribe liver ultrasonography every 6 months to these patients. Metastatic tumors can occur in the uvea and more frequently in the posterior part called the choroid. They are more frequent after breast cancer and lung cancer. Conjunctival tumors can be epithelial (benign papillomas and epidermoid carcinomas) or melanocytic (benign naevi and melanomas) and sometimes from sebaceous gland origin (sebaceous carcinomas). Conjunctival malignancies can metastasize to the regional lymph nodes and other organs. All these tumors are rare tumors and need to be managed in expert centers. We describe the psychological consequences and the French national network organization for treatment of uveal malignant melanoma.

Résumé:
Nous décrivons les principales tumeurs malignes intraoculaires, les tumeurs conjonctivales et citons également quelques tumeurs orbitopalpébrales. Les tumeurs malignes primitives intraoculaires sont le rétinoblastome chez l’enfant et le mélanome uvéal chez l’adulte. Le mélanome uvéal métastase essentiellement au niveau du foie ce qui justifie une surveillance hépatique régulière. Les métastases peuvent survenir dans l’uvée également le plus souvent l’uvée postérieure ou choroïde et sont plus fréquentes après cancer du sein ou du poumon. Les tumeurs conjonctivales peuvent être épithéliales (papillome bénins et carcinome épidermoïdes malins in situ ou invasifs) ou mélanocytaires (nævi bénins et mélanome malins) voire parfois sébacées. Les tumeurs malignes invasives peuvent envahir les aires ganglionnaires et métastaser. Toutes ces tumeurs sont des tumeurs rares qui doivent être pris e en charge dans des centres experts. Nous décrivons leur retentissement psychologique et le réseau national de prise en charge du mélanome uvéal.     

外周血流式细胞仪分析在淋巴瘤性红皮病诊断中的作用研究

目的:探讨外周血流式细胞仪检测在红皮病诊断中的应用价值。方法:收集2017年9月至2020年12月在中国医学科学院皮肤病医院就诊的29例红皮病患者,包括6例红皮病型蕈样肉芽肿（EMF）、5例Sézary综合征（SS）及18例不同病因的炎症性红皮病（IE）,4例健康志愿者为健康对照。采用流式细胞仪检测外周血淋巴细胞亚群、...     

骨盆三联截骨联合股骨近端截骨术治疗Legg-Calvé-Perthes 病的疗效

目的：评价骨盆三联截骨联合股骨近端截骨术治疗Legg‐Calvé‐Perthes病的短期疗效。方法回顾性分析2010年5月至2012年12月采用包容治疗的40例（40髋）Legg‐Calvé‐Perthes病患者资料,其中男29例,女11例;年龄6岁～10岁4个月,平均8．3岁。根据包容治疗方法将患者分为2组：A组,20例,采用骨盆三联截骨联合股骨近端截骨术;B组：20例,采用其他包容治疗（股骨近端截骨术、Salter骨盆截骨联合股骨近端截骨术）。比较两组手术前后影像学数据变化和 Harris髋关节评分。结果所有患者均获得术后18～48个月（平均32个月）的随访。术后 A、B组C E角分别增加18．45°±4．25°和13．18°±3．07°,股骨头外移比值分别降低0．53±0．25和0．35±0．19,Sharp角分别降低14．64°±3．25°和10．87°±6．42°,股骨头覆盖率分别增加22．42％±2．19％和18．57％±6．42％,差异均有统计学意义（P＜0．05）。A组术后Harris髋关节评分4分及以上19例,B组18例,差异无统计学意义（P＞0．05）。结论骨盆三联截骨联合股骨近端截骨术治疗Legg‐Calvé‐Perthes病髋关节局部解剖结构改变较小,能达到很好的包容效果,可有效防止单一术式局部矫正过大引发的并发症,适用于严重Legg‐Calvé‐Perthes病。     

利用RAPD分子标记检测空间飞行诱导的蕃茄DNA突变

为快速、有效地分析经空间飞行的蕃茄基因组ＤＮＡ，采用了随机引物扩增多态ＤＮＡ分析的分子标记方法，检测了基因序列的多态性。蕃茄种子搭载返地卫星，空间飞行３５５ｈ，回收后地面种植，选择三个同工酶有差异的空间处理植株进行ＲＡＰＤ检测。结果表明：在５０个供试引物中，有１８个扩增出了ＤＮＡ带，共有１５６条，其大小在２００￣２０００ｂｐ之间。