利用四维锥形束CT指导肺癌内靶区的勾画

目的 研究四维锥形束CT(4D-CBCT)指导肺癌内靶区勾画的可行性。方法 简单随机法选取本院24例肺癌患者。平静呼吸下CT模拟定位获得CT图像,首次治疗前4D-CBCT扫描获得4D-CBCT中位图像,将4D图像重建算法更改为3D,获得3D-CBCT图像。图像融合算法取骨配准,分别在定位CT、4D-CBCT中位图像、3D-CBCT图像上勾画大体肿瘤靶区(GTV),定义GTV到临床靶区(CTV)的外扩边界为7 mm,获得CTV_CT、ITV_4D和ITV_3D,基于CTV_CT在三维方向上外扩5 mm得到ITV_CT。比较靶区间中心点位置、体积、相似度和相互包含关系的差异。结果 ITV_CT与ITV_3D在中下叶组的头脚方向上,靶区中心点位置差异有统计学意义(Z=-2.027,P<0.05)。在靶区体积方面,ITV_CT最大,与ITV_3D相比差异有统计学意义(Z=-2.941,P<0.05),ITV_4D最小,但与ITV_3D相比差异无统计学意义(P>0.05)。ITV_CT与ITV_3D相似度均数<75%,ITV_4D与ITV_3D相似度均数>90%(Z=-2.940、-2.975,P<0.05)。ITV_CT、ITV_4D未被ITV_3D包含的比例均数为40%和5%(Z=-2.952、-3.185,P<0.05)。结论 4D-CBCT的中位图像可以缩小内靶区的勾画范围,为肺癌的自适应放疗提供选择。     

Sellar chordoma presenting as pseudo-macroprolactinoma with unilateral third cranial nerve palsy

We described a 61‐year‐old female with a sellar chordoma, which presented as pseudo‐macroprolactinoma with unilateral third cranial nerve palsy. Physical examination revealed that her right upper lid could not be raised by itself, right eyeball movement limited to the abduction direction, right pupil dilated to 4.5 mm with negative reaction to light, and hemianopsia in bitemporal sides. CT scanning showed a hyperdense lesion at sellar region without bone destruction. Magnetic resonance imaging (MRI) revealed the tumor was 2.3 cm×1.8 cm×2.6 cm, with iso‐intensity on T1WI, hyper‐intensity on T2WI and heterogeneous enhancement on contrast imaging. Endocrine examination showed her serum prolactin level increased to 1,031.49 mIU/ml. The tumor was sub‐totally resected via pterional craniotomy under microscope and was histologically proven to be a chordoma. Postoperatively, she recovered uneventfully but ptosis and hemianopsia remained at the 6th month.     

Novel mutations in PDE6B causing human retinitis pigmentosa

AIM: To identify the genetic defects of a Chinese patient with sporadic retinitis pigmentosa (RP). METHODS: Ophthalmologic examinations were performed on the sporadic RP patient, 144 genes associated with retinal diseases were scanned with capture next generation sequencing (CNGS) approach. Two heterozygous mutations in PDE6B were confirmed in the pedigree by Sanger sequencing subsequently. The carrier frequency of PDE6B mutations of reported PDE6B mutations based on the available two public exome databases (1000 Genomes Project and ESP6500 Genomes Project) and one in-house exome database was investigated. RESULTS: We identified compound heterozygosity of two novel nonsense mutations c.1133G>A (p.W378X) and c.2395C>T (p.R799X) in PDE6B, one reported causative gene for RP. Neither of the two mutations in our study was presented in three exome databases. Two mutations (p.R74C and p.T604I) in PDE6B have relatively high frequencies in the ESP6500 and in-house databases, respectively, while no common dominant mutation in each of the database or across all databases. CONCLUSION: We demonstrates that compound heterozygosity of two novel nonsense mutations in PDE6B could lead to RP. These results collectively point to enormous potential of next-generation sequencing in determining the genetic etiology of RP and how various mutations in PDE6B contribute to the genetic heterogeneity of RP.     

抗击严重急性呼吸综合征的医生与护士心理状态的比较与分析

为分析抗击严重急性呼吸综合征(SARS)的医生与护士的心理状态和差异,在SARS暴发的3个月中共收集了来自多家SARS定点医院的1532份医生和护士的心理健康问卷,应用时勘心理量表与Zung自评抑郁量表进行分析比较。发现无论处于一线还是二线,医生的一般心理健康得分要高于护士,自评抑郁量表得分低于护士,且护士的自评抑郁量表得分要高于常模组。逐步相关回归分析显示,抑郁量表得分同压力源,如“社会对医护人员的支持不够”,“治疗条件和生活环境不好”,“缺乏家庭亲人的支持和交流”等有关,同参加体育锻炼的程度,文化程度呈负相关。医生的心理状态和抑郁状态好于护士。提示应针对不同原因给予干预,同时应对于发现的相关因素予以改善。     

LncRNA UCA1对肺癌细胞放射敏感性影响及机制研究

目的 研究长链非编码RNA (LncRNA) UCA1对肺癌细胞增殖、凋亡及放射敏感性影响及其机制。方法 运用qRT-PCR法检测肺癌细胞A549、H1299和人正常肺细胞HBE中UCA1、miR-513a-5p表达。将si-con组(转染si-con)、si-UCA1组(转染si-UCA1)、miR-513a-5p组(转染miR-513a-5p mimics)、miR-NC组(转染miR-NC)、IR+si-con组(转染si-con+照射)、IR+si-UCA1组(转染miR-NC+照射)、IR+miR-513a-5p组(转染miR-513a-5p mimics+照射)、IR+miR-NC组(转染miR-NC+照射)、IR+si-UCA1+anti-miR-513a-5p组(共转染si-UCA1和anti-miR-513a-5p+照射)均用脂质体法转染至A549、H1299细胞,然后部分组进行4Gy照射。MTT法检测各组细胞增殖,克隆形成实验检测细胞增敏比,流式细胞术检测各组细胞凋亡,双荧光素没报告基因检测实验检测各组细胞的荧光活性。结果 与HBE细胞相比,A549、H1299细胞中UCA1表达显著升高(P<0.05),miR-513a-5p表达显著降低(P<0.05)。抑制UCA1、过表达miR-513a-5p均可明显抑制A549、H1299细胞增殖、促进凋亡、提高放射敏感性(放射增敏比为1.897、2.146和1.615、1.872)。miR-513a-5p可抑制野生型UCA1细胞的荧光活性,且UCA1可负向调控miR-513a-5p的表达。抑制miR-513a-5p可逆转抑制UCA1对细胞的放射敏感性的增强作用。结论 抑制LncRNA UCA1可增强放射对肺癌细胞敏感性,其机制可能与靶向抑制miR-513a-5p有关。     

HPLC-ELSD法测定贝母中异甾类生物碱及糖苷类成分的含量

目的建立同时测定贝母中5种异甾类生物碱——peimissine, imperialine, sinpeinine A, imperialine-3β-glucoside和yibeinoside A含量的HPLC分析方法。方法C₁₈柱;流动相:乙腈-水(含0.1%二乙胺);梯度洗脱,流速1.0 mL·min^-1;检测器:Alltech 500蒸发光散射检测器(ELSD)。结果线性范围为peimissine 13.1～288.2 mg·L^-1(r²=0.997 5), imperialine-3β-glucoside 7.7～169.4 mg·L^-1 (r²=0.999 3), yibeinoside A 7.3～160.6 mg·L^-1 (r²=0.999 7), imperialine 16.5～363.0 mg·L^-1 (r²=0.999 2), sinpeinine A 8.7～191.4 mg·L^-1 (r²=0.994 2)。 5个化合物的精密度和重现性RSD均<5%。结论本方法简便、有效、可行,可用于贝母中5种异甾类生物碱的含量测定。     

Circ_0008039 supports breast cancer cell proliferation,migration, invasion,and glycolysis by regulating the miR‐140‐3p/SKA2 axis

Circular RNAs (circRNAs) have been shown to modulate gene expression and participate in the development of multiple malignancies. The purpose of this study was to investigate the role of circ_0008039 in breast cancer (BC). The expression of circ_0008039, miR‐140‐3p, and spindle and kinetochore‐associated protein 2 (SKA2) was detected by qRT‐PCR. Cell viability, colony formation, migration, and invasion were evaluated using methylthiazolyldiphenyl‐tetrazolium bromide (MTT) assay, colony formation assay, and transwell assay, respectively. Glucose consumption and lactate production were measured using commercial kits. Protein levels of hexokinase II (HK2) and SKA2 were determined by western blot. The interaction between miR‐140‐3p and circ_0008039 or SKA2 was verified by dual‐luciferase reporter assay. Finally, a mouse xenograft model was established to investigate the roles of circ_0008039 in BC in vivo. We found that circ_0008039 and SKA2 were upregulated in BC tissues and cells, while miR‐140‐3p was downregulated. Knockdown of circ_0008039 suppressed BC cell proliferation, migration, invasion, and glycolysis. Moreover, miR‐140‐3p could bind to circ_0008039 and its inhibition reversed the inhibitory effect of circ_0008039 interference on proliferation, migration, invasion, and glycolysis in BC cells. SKA2 was verified as a direct target of miR‐140‐3p and its overexpression partially inhibited the suppressive effect of miR‐140‐3p restoration in BC cells. Additionally, circ_0008039 positively regulated SKA2 expression by sponging miR‐140‐3p. Consistently, silencing circ_0008039 restrained tumor growth via increasing miR‐140‐3p and decreasing SKA2. In conclusion, circ_0008039 downregulation suppressed BC cell proliferation, migration, invasion, and glycolysis partially through regulating the miR‐140‐3p/SKA2 axis, providing an important theoretical basis for treatment of BC.

Abbreviations

ANOVA

analysis of variance

BC

breast cancer

circRNAs

circular RNAs

DMSO

dimethyl sulfoxide

ECAR

extracellular acidification rate

ECL

enhanced chemiluminescence

FBS

fetal bovine serum

HK2

hexokinase II

MEGM

mammary epithelial growth medium

miR‐140‐3p

microRNA‐140‐3p

MTT

methylthiazolyldiphenyl‐tetrazolium bromide

PBS

phosphate‐buffered saline

PRKAR1B

protein kinase A regulatory subunit R1‐beta

SD

standard ± deviation

SKA2

spindle and kinetochore‐associated protein 2

     

Effect of electroacupuncture on glial fibrillary acidic protein and nerve growth factor in the hippocampus of rats with hyperlipidemia and middle cerebral artery thrombus

Electroacupuncture (EA) has been shown to reduce blood lipid level and improve cerebral ischemia in rats with hyperlipemia complicated by cerebral ischemia. However, there are few studies on the results and mechanism of the effect of EA in reducing blood lipid level or promoting neural repair after stroke in hyperlipidemic subjects. In this study, EA was applied to a rat model of hyperlipidemia and middle cerebral artery thrombosis and the condition of neurons and astrocytes after hippocampal injury was assessed. Except for the normal group, rats in other groups were fed a high-fat diet throughout the whole experiment. Hyperlipidemia models were established in rats fed a high-fat diet for 6 weeks. Middle cerebral artery thrombus models were induced by pasting 50% FeCl₃ filter paper on the left middle cerebral artery for 20 minutes on day 50 as the model group. EA1 group rats received EA at bilateral ST40 (Fenglong) for 7 days before the thrombosis. Rats in the EA1 and EA2 groups received EA at GV20 (Baihui) and bilateral ST40 for 14 days after model establishment. Neuronal health was assessed by hematoxylin-eosin staining in the brain. Hyperlipidemia was assessed by biochemical methods that measured total cholesterol, triglyceride, low-density lipoprotein and high-density lipoprotein in blood sera. Behavioral analysis was used to confirm the establishment of the model. Immunohistochemical methods were used to detect the expression of glial fibrillary acidic protein and nerve growth factor in the hippocampal CA1 region. The results demonstrated that, compared with the model group, blood lipid levels significantly decreased, glial fibrillary acidic protein immunoreactivity was significantly weakened and nerve growth factor immunoreactivity was significantly enhanced in the EA1 and EA2 groups. The repair effect was superior in the EA1 group than in the EA2 group. These findings confirm that EA can reduce blood lipid, inhibit glial fibrillary acidic protein expression and promote nerve growth factor expression in the hippocampal CA1 region after hyperlipidemia and middle cerebral artery thrombosis. All experimental procedures and protocols were approved by the Animal Use and Management Committee of Beijing University of Chinese Medicine, China (approval No. BUCM-3-2018022802-1002) on April 12, 2018.

Chinese Library Classification No. R459.9; R363; R364     

NRAGE影响人食管癌细胞放射抗性的作用及其机制研究

背景与目的：食管癌是全球威胁人类生命和健康的常见恶性肿瘤之一，中国每年食管癌发病人数占全球发病总人数的一半以上，以食管鳞癌最为常见。放疗是食管癌三大治疗手段之一，而放射抗性是导致其治疗失败的主要原因。神经营养因子受体相互作用MAGE类药物（neurotrophin receptor-interacting MAGE homolog，NRAGE）在放射抗性细胞株TE13R120中表达量明显高于亲本TE13细胞，且NRAGE亚细胞定位变化可能参与食管癌细胞放射抗性的形成。通过基因转染构建NRAGE稳定表达的食管癌细胞系，以进一步明确NRAGE基因与食管鳞癌细胞放射抗性的关系，分析该基因影响放射抗性的具体机制。方法：通过基因转染构建NRAGE稳定表达的食管癌细胞系。采用细胞克隆形成实验检测细胞放射敏感性，采用流式细胞术检测细胞周期及凋亡；细胞划痕、Transwell侵袭实验检测细胞迁移、侵袭能力，采用实时荧光定量聚合酶链反应（real-time fluorescence quantitive polymerase chain reaction，RTFQ-PCR）和蛋白质印迹法（Western blot）检测细胞中β-catenin表达情况。组间差异采用t检验或方差分析。结果：实验分为转染过表达组（Eca109/NRAGE组）和空白对照组（Eca109组）。Eca109/NRAGE细胞中NRAGE的表达量明显高于Eca109（t=29.65，P<0.05）。照射后Eca109/NRAGE细胞的放射抗性显著高于Eca109。流式细胞术检测结果显示，Eca109/NRAGE细胞中对射线抵抗性最强的S期细胞比例增加，对射线最敏感的G ₂ /M期减少。Eca109/NRAGE细胞凋亡率较Eca109细胞降低（t=3.268，P<0.05）。Eca109/NRAGE细胞迁移和侵袭能力均高于Eca109。RTFQ-PCR和Western blot检测结果显示，β-catenin的mRNA表达及蛋白水平在Eca109/NRAGE细胞中明显高于Eca109（t=15.87，P<0.05）。结论：NRAGE参与Eca109细胞放射抗性的形成，改变细胞的细胞周期分布和凋亡情况，影响细胞迁移及侵袭能力并可能影响食管癌细胞的放射敏感性，该作用可能与激活Wnt/β-catenin信号转导通路有关。