Saikosaponin-d increases radiation-induced apoptosis of hepatoma cells by promoting autophagy via inhibiting mTOR phosphorylation

Objective: The aim of this study was to analyze the effects of saikosaponin-d (SSd) on autophagy activity and radiosensitivity of hepatoma cells, and to elucidate its related molecular mechanisms.Methods: The growth of SMMC-7721 and MHCC97L hepatoma cells were detected by clonal formation and survival fraction. Flow cytometry was used to detect the changes of apoptosis of hepatoma cells. The morphological changes of autophagy of hepatoma cells were observed by transmission electron microscopy and were further quantitatively detected by laser confocal microscopy. The expressions of related proteins were detected by Western blotting.Results: SSd can significantly increase the apoptosis of hepatoma cells induced by radiation and inhibit the proliferation of hepatoma cells. The addition of the autophagy inhibitor chloroquine (CQ) or an mTOR agonist (MHY1485), which could reduce the promoting effect of SSd on radiation-induced apoptosis and inhibitory effect on the proliferation of hepatoma cells. Transmission electron microscopy and confocal microscopy results also showed that the number of autophagosomes was significantly higher in the radiation and SSd co-treatment group than in the radiotherapy or SSd alone group; however, the effect of SSd on autophagy in hepatoma cells was decreased after adding MHY1485, siRNA-P53 or AMPK inhibitor (Compound C). Western blot analysis showed that after the addition of SSd, the phosphorylation of mTOR was significantly decreased by radiation, the expression of the autophagy-related proteins LC3-II and Beclin-1 was increased, p62 was decreased, and the expression of cleaved caspase-3 and cleaved PARP was enhanced; this effect of SSd was partially reversed after the addition of MHY1485, siRNA-P53 or Compound C.Conclusions: SSd increases radiation-induced apoptosis of hepatoma cells by promoting autophagy via inhibiting mTOR phosphorylation and providing a possible potential approach for radiosensitization therapy of liver cancer.     

Timing of dental extractions in patients undergoing radiotherapy and the incidence of osteoradionecrosis: a systematic review and meta-analysis

This systematic review aimed to examine whether the incidence of osteonecrosis differed between patients who have dental extractions before or after radiotherapy (RT). The reported incidence of osteoradionecrosis (ORN) of the jaws following RT to the head and neck varies widely in the literature. Currently, for patients with head and neck cancer there are no universally accepted guidelines on the optimal timing of dental surgery relative to RT to minimise incident ORN. A literature review was conducted using the Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) criteria. A search of PubMed, EMBASE, Evidence-Based Medicine, and Web of Science databases targeted literature published up to and including 10 April 2020. Two independent reviewers assessed studies for eligibility against inclusion criteria. An assessment of bias was conducted for each of the included studies and relevant data extracted. A meta-analysis was undertaken using the statistical methods described. Twenty-four of 708 studies were included. They were heterogeneous and included a wide variation of RT methods, head and neck malignancies, and comorbidities. While some concluded that the incidence of ORN was dependent on the timing of dental extractions in relation to RT, with regard to the risk of its development, others reported additional factors such as age, comorbidities, extent of surgical resection, and dose and field of radiation, as more important predictors than timing. In many there was consistent lack of detail around the timing of dental procedures in relation to the delivery of RT. From 21 studies including 36,294 patients, of whom 14,389 had extractions before RT, the pooled incidence of ORN was 5.5% (95% CI: 2.1% to 10.1%). Significant heterogeneity was found in Cochran’s Q-test (p < 0.001) and Higgins I² = 98.0%. From 21 studies including 37,805 patients, of whom 6030 had extractions after RT, the pooled incidence of ORN was 5.3% (95% CI: 2.9% to 8.2%). Significant heterogeneity was found in Cochran’s Q-test (p < 0.001) and Higgins I² = 80.0%. There was no statistically significant difference between these two groups (random-effects model Q=0.12, p=0.73). Large, longitudinal studies with a priori-specified methods are needed to identify, recruit, and prospectively follow patients with head and neck cancer for the onset of ORN after dental surgery. This will allow clinical guidelines to be established to assist clinicians to plan treatment when extractions are indicated in patients undergoing RT to the head and neck.     

Assessment of IGRT variability for lung SBRT

PurposeThe purpose of this project was to assess factors that may influence variability in the pre-treatment kilovoltage cone beam computed tomography (kV CBCT) image matching process for lung stereotactic body radiation therapy (SBRT).Methods and materialsPre-treatment CBCT and planning CT data sets of previously-treated lung SBRT patients were gathered and anonymized from four radiotherapy centers in Alberta. Eight radiation therapists (RTTs) and four radiation oncologists (ROs) were recruited from the same four cancer centers for image matching. Identical data sets were provided to each user, but the order of image sets was randomized independently for each user to remove any learning bias. Inter-user variabilities were then investigated as functions of various factors, including image origin (source institution/machine), user's institution (local matching protocol), profession (RTT vs. RO), years of experience and image quality (presence/absence of added noise).ResultsVery little variation in image matching between different users was observed. The mean differences from the consensus means for different image sets were less than 1 mm in all directions, and cases that exceeded 3 mm (i.e. clinically significant differences) were extremely rare. Image origin, user's institution, and profession (RTT vs. RO) didn't lead to any meaningful clinical differences, while image quality didn't introduce any statistically significant differences. In addition, no discernible trend was seen between user's experience and deviation from the user mean. Overall, no meaningful differences in inter-user variabilities for the different factors investigated were found in this study.ConclusionsThere appears to be an adequate standardization across the province of Alberta in terms of CBCT image matching process. No clinically significant differences were observed as functions of various factors investigated in this study. Consistency in matching between RTTs and ROs in this study suggests that RTTs do not need systematic RO approval of their lung CBCT match. It should be noted that RTTs at the centers in this study receive comprehensive training in CBCT-based image matching.     

Protective effects of a cystine and theanine mixture against acute radiation injury in rats

This study aims to examine the effects of cystine and theanine (CT), which increases glutathione biosynthesis, on the survival rate and acute radiation injury of the small intestine and bone marrow using a rat model. CT pre-treatment (280 mg/kg for 5 days) significantly improved weight loss and survival rate of rats as compared with the control group after 5 Gy. CT pre-treatment significantly increased the rate of mucosa and crypt length, and decreased the number of apoptotic cells, TUNEL and cleaved caspase-3 positive cells, while increasing the number of mitotic cells and Ki-67 positive cells in jejunal crypts and villi compared to control rats post-irradiation. CT also suppressed bone marrow cell loss and reduced the number of apoptotic cells in bone marrow. These results suggest a protective effect of CT pre-treatment for acute injury after irradiation through apoptosis inhibition and increased proliferative activity in jejunal crypt cells and bone marrow cells.     

腹盆腔放疗诱发肠道微生态失调与肠源性感染的实验研究

目的 探索腹盆腔放疗照射对肠道微生态的影响及其与肠源性感染的关系。方法 模拟腹盆腔放疗照射BALB/c小鼠,2.0 Gy/d,连续照射5 d/周,分别于照射3周、5周和6周后停照1周的时间点收集回肠组织及其内容物样本。用实时定量RT-PCR检测抗菌肽和促炎性因子的表达;用PCR检测细菌在小鼠体内的移位情况;用变性梯度凝胶电泳技术检测分析肠道微生态的特征。结果 腹盆腔照射诱发了肠道潘氏细胞隐窝素-1和-4表达紊乱,照射3周或照射6周后停照1周,小鼠回肠隐窝素-1和-4均呈现显著性降低(t=-7.43、-3.54、-4.72、-4.27,P<0.05);而照射5周小鼠回肠隐窝素-1和-4表达明显升高(t=6.15、5.75,P<0.05)。放疗模拟照射3和5周时小鼠肠道微生物区系多样性指数和丰富度显著降低(t=-3.49、-4.19、-3.44、-4.97,P<0.05),呈现以乳酸杆菌等益生菌减少,大肠杆菌和弗氏志贺氏菌等条件致病菌增多为特征的微生态失调。受照小鼠肠系膜淋巴结和血液中的细菌DNA阳性率明显增高。照射3和5周后回肠组织IL-1β、IL-6和TNF-α显著性高表达(t=4.85、6.16、7.71、4.60、4.86、5.97,P<0.05);照射6周后停照1周时,肠道促炎性因子的表达量有所回落,但IL-1β和TNF-α的表达量仍显著性高表达(t=3.67、5.88,P<0.05)。结论 腹盆腔放疗可诱发肠道抗菌肽表达紊乱,引起肠道微生态失调,进而导致肠源性细菌移位及感染性炎症的发生。微生态可能成为减轻放疗患者消化道不良反应的有效干预靶点。     

γ刀照射三叉神经的剂量-效应关系研究

目的 探讨伽玛刀不同剂量照射大鼠三叉神经(感觉根)后对照射范围内三叉神经组织的损伤以及对周围临近脑干组织和脑桥的影响,为优化伽玛刀照射剂量提供理论依据。方法 选用成年雄性SD大鼠65只,分别以不同照射剂量(20、40、80、160 Gy)照射三叉神经,观察照射后1周、2周、4周大鼠行为学变化,取全脑组织进行HE、Nissl、MAPLC3B、53BP1、XRCC1染色观察细胞形态学和自噬以及DNA损伤和修复等变化。结果 所有剂量组照射大鼠均存活。20和40 Gy组均无行为学和病理学改变;80和160 Gy组均出现了照射部位毛发脱落;HE、Nissle、LC3B、53BP1及XRCC1染色发现,受照部位组织在80和160 Gy照射后,可见神经元细胞固缩、膨胀,神经元细胞发生自噬以及DNA损伤明显,160 Gy组DNA损伤明显加重。40 Gy照射后,神经元发生DNA损伤修复的程度较其他剂量增高。结论 伽玛刀照射三叉神经根对神经元有明显的剂量-效应关系。     

低管电压联合迭代模型重建技术在肝脏CT增强扫描中的可行性研究

目的 探讨低管电压联合迭代模型重建(IMR)技术在肝脏CT增强扫描中的可行性。方法 60例患者按随机数字表法分为A组和B组,每组30例。扫描方案A组动脉期100 kV,门静脉期120 kV,B组动脉期120 kV,门静脉期100 kV。各组管电流均固定为250 mAs。A组动脉期和B组门静脉期采用IMR,A组门静脉期和B组动脉期采用滤波反投影(FBP)重建,得到4组图像,包括A1组(动脉期,100 kV,IMR),B1组(动脉期,120 kV,FBP),A2组(门静脉期,120 kV,FBP)以及B2组(门静脉期,100 kV,IMR)。分别比较A1组和B1组,A2组和B2组的图像质量客观评价指标 [图像噪声、图像信噪比(SNR)、对比噪声比(CNR)] 和主观评价指标(低对比分辨力、病灶边缘锐利度、图像失真及诊断信心度),并计算有效剂量。结果 有效剂量A1组较B1组、B2组较A2组明显下降(t=11.05、11.64, P<0.01)。低对比分辨力、病灶边缘锐利度A1优于B1组、 B2优于A2组(Z=6.391、3.200、6.559、3.409, P<0.01),图像失真和诊断信心差异无统计学意义(P>0.05)。图像噪声A1组低于B1组,B2组低于A2组(t=12.889、15.163, P<0.01),SNR和CNR A1组高于B1组,B2组高于A2组(t=15.458、1.325、15.308、3.136, P<0.01)。结论 与常规管电压FBP重建相比,低管电压联合IMR重建可显著降低肝脏增强CT的辐射剂量,并提高其图像质量。     

A multidisciplinary approach unravels early and persistent effects of X-ray exposure at the onset of prenatal neurogenesis

Background

In humans, in utero exposure to ionising radiation results in an increased prevalence of neurological aberrations, such as small head size, mental retardation and decreased IQ levels. Yet, the association between early damaging events and long-term neuronal anomalies remains largely elusive.

Methods

Mice were exposed to different X-ray doses, ranging between 0.0 and 1.0 Gy, at embryonic days (E) 10, 11 or 12 and subjected to behavioural tests at 12 weeks of age. Underlying mechanisms of irradiation at E11 were further unravelled using magnetic resonance imaging (MRI) and spectroscopy, diffusion tensor imaging, gene expression profiling, histology and immunohistochemistry.

Results

Irradiation at the onset of neurogenesis elicited behavioural changes in young adult mice, dependent on the timing of exposure. As locomotor behaviour and hippocampal-dependent spatial learning and memory were most particularly affected after irradiation at E11 with 1.0 Gy, this condition was used for further mechanistic analyses, focusing on the cerebral cortex and hippocampus. A classical p53-mediated apoptotic response was found shortly after exposure. Strikingly, in the neocortex, the majority of apoptotic and microglial cells were residing in the outer layer at 24 h after irradiation, suggesting cell death occurrence in differentiating neurons rather than proliferating cells. Furthermore, total brain volume, cortical thickness and ventricle size were decreased in the irradiated embryos. At 40 weeks of age, MRI showed that the ventricles were enlarged whereas N-acetyl aspartate concentrations and functional anisotropy were reduced in the cortex of the irradiated animals, indicating a decrease in neuronal cell number and persistent neuroinflammation. Finally, in the hippocampus, we revealed a reduction in general neurogenic proliferation and in the amount of Sox2-positive precursors after radiation exposure, although only at a juvenile age.

Conclusions

Our findings provide evidence for a radiation-induced disruption of mouse brain development, resulting in behavioural differences. We propose that alterations in cortical morphology and juvenile hippocampal neurogenesis might both contribute to the observed aberrant behaviour. Furthermore, our results challenge the generally assumed view of a higher radiosensitivity in dividing cells. Overall, this study offers new insights into irradiation-dependent effects in the embryonic brain, of relevance for the neurodevelopmental and radiobiological field.

Electronic supplementary material

The online version of this article (doi:10.1186/1866-1955-7-3) contains supplementary material, which is available to authorized users.     

A simple and effective treatment for hemorrhagic radiation proctitis using formalin

Radiation proctitis is a common complication of radiotherapy for pelvic malignancy. In the more severe form, it leads to intractable or massive hemorrhage, which may require repeated hospital admissions and blood transfusions. Medical therapy in patients with radiation proctitis is usually ineffective, whereas surgery is associated with a high morbidity and mortality. Eight patients (seven females and one male) with hemorrhagic radiation proctitis were Treated over a six-month period with endoluminal formalin. The technique used ensured minimal contact with formalin. The median age of the patients was 68 years (range, 42–73 years). Seven patients had had cancer of the uterine cervix, and one patient had had cancer of the prostate treated with radiotherapy at a median time of 30 months (range, 9–46 months) previously. The median duration of time of symptomatic rectal hemorrhage before formalin therapy was eight months (range, 1–12 months). The median number of units of blood transfused previously per patient was four (range, 2–32). The time taken for formalin therapy was 20 minutes (range, 10–70 minutes). One patient required repeat formalin application at two weeks. Bleeding ceased immediately in seven patients after formalin treatment. No further bleeding was noted, nor was any blood transfusion needed, at follow-up at four months (range, 1–6 months). Formalin therapy is a simple, inexpensive, and effective treatment for hemorrhagic radiation proctitis.