The intergenerational legacy of the 1959-1961 Great Chinese Famine on children's cognitive development.

We investigate the effect of early exposure to malnutrition on the cognitive abilities of the offspring of survivors in the context of a natural experiment; i.e., the Great Chinese Famine (GCF) of 1959-61. We employ a novel dataset - the China Family Panel Studies (CFPS) - to do so. The paper finds that the cognitive abilities of children whose fathers were born in rural areas during the famine years (1959-1961) were impaired by exposure to the GCF and the negative effect was greater for girls than boys, whereas children whose mothers were born in rural areas during the famine years were not affected. The uncovered gender-specific effect is almost entirely attributable to son preference exhibited in families with male famine survivors.

Advantages and robustness of partial VMAT with prone position for neoadjuvant rectal cancer evaluated by CBCT-based offline adaptive radiotherapy.

BACKGROUND AND PURPOSE: This study aims to explore the advantages and robustness of the partial arc combined with prone position planning technique for radiotherapy in rectal cancer patients. Adaptive radiotherapy is recalculated and accumulated on the synthesis CT (sCT) obtained by deformable image registration between planning CT and cone beam CT (CBCT). Full and partial volume modulation arc therapy (VMAT) with the prone position on gastrointestinal and urogenital toxicity, based on the probability of normal tissue complications (NTCP) model in rectal cancer patients were evaluated. MATERIALS AND METHODS: Thirty-one patients were studied retrospectively. The contours of different structures were outlined in 155 CBCT images. First, full VMAT (F-VMAT) and partial VMAT (P-VMAT) planning techniques were designed and calculated using the same optimization constraints for each individual patient. The Acuros XB (AXB) algorithm was used in order to generate more realistic dose distributions and DVH, considering the air cavities. Second, the Velocity 4.0 software was used to fuse the planning CT and CBCT to obtain the sCT. Then, the AXB algorithm was used in the Eclipse 15.6 software to conduct re-calculation based on the sCT to obtain the corresponding dose. Furthermore, the NTCP model was used to analyze its radiobiological side effects on the bladder and the bowel bag. RESULTS: With a CTV coverage of 98%, when compared with F-VMAT, P-VMAT with the prone position technique can effectively reduce the mean dose of the bladder and the bowel bag. The NTCP model showed that the P-VMAT combined with the prone planning technique resulted in a significantly lower complication probability of the bladder (1.88 ± 2.08 vs 1.62 ± 1.41, P = 0.041) and the bowel bag (1.28 ± 1.70 vs 0.95 ± 1.52, P < 0.001) than the F-VMAT. In terms of robustness, P-VMAT was more robust than F-VMAT, considering that less dose and NTCP variation was observed in the CTV, bladder and bowel bag. CONCLUSION: This study analyzed the advantages and robustness of the P-VMAT in the prone position from three aspects, based on the sCT fused by CBCT. Whether it is in regards to dosimetry, radiobiological effects or robustness, P-VMAT in the prone position has shown comparative advantages.

RUNX2 promotes gastric cancer progression through the transcriptional activation of MGAT5 and MMP13.

Introduction: RUNX2 is overexpressed in gastric cancer but the mechanism(s) through which it promotes tumor progression remain undefined. Here, we investigated the role of RUNX2 on gastric cancer pathogenesis at the molecular level. Methods: The qRT-PCR and western bolt were utilized to examine the mRNA and protein levels. CCK-8, Transwell and wound healing assays were used to measure cell proliferation, invasion and migration. CHIP-PCR gel electrophoresis was used to verify RUNX2 as a transcription factor for MMP13 and MGAT5. The in vivo assay was utilized to assess tumor growth. In vivo assay was used to evaluate tumor growth, aberrant expression of RUNX2 and lung metastasis of gastric cancer. Results: RUNX2 is overexpressed in MKN-45 and AGS cells. Genetic RUNX2 silencing reduced the proliferation, invasion and migration of MKN-45 and AGS cells. Analysis of the gastric cancer samples from the database revealed a significant positive correlation between MGAT5, MMP13, and RUNX2 expression. JASPAR analysis revealed that there was a potential binding site of RUNX2 in the promoter regions of MGAT5 and MMP13, and the experimental results confirmed that RUNX2 could regulate the expression of MGAT5 and MMP13 respectively. In vivo assays confirmed the aberrant expression of RUNX2 in mouse models of gastric cancer and reduced growth and lung metastasis in RUNX2 silenced xenograft tumors assessed. Conclusion: Collectively, these data reveal that RUNX2 enhances MGAT5 and MMP13 expression in gastric cancer cells and represents a biomarker and potential therapeutic target for gastric cancer therapy.

E2F1 as a potential prognostic and therapeutic biomarker by affecting tumor development and immune microenvironment in hepatocellular carcinoma.

Background: E2F1 plays a crucial role in cell cycle regulation. However, the exact role of E2F1 in liver hepatocellular carcinoma (LIHC) remains controversial. This study aimed to integrate disparate data by bioinformatics for a deeper insight into the possible roles of E2F1 in LIHC. Methods: Differentially overexpressed genes in LIHC were screened by GEO2R. Gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were analyzed by WebGestalt. Then, hub genes were selected via STRING and Cytoscape, followed by validation with Oncomine, GEPIA2, and Human Protein Atlas (HPA). Next, E2F1 expression was investigated using Oncomine, GEPIA2, TIMER2.0, UALCAN, and HPA. Then, Kaplan-Meier plotter was adopted to investigate survival. After that, E2F1 promotor methylation, mutations and copy number alterations were analyzed with UALCAN and cBioPortal. Moreover, competing endogenous RNAs (ceRNAs) network were established using ENCORI, miRCancer, and Kaplan-Meier plotter. Additionally, the association between E2F1 and immune microenvironment was investigated through TISCH and TIMER2.0. Results: Six hub genes including E2F1 were identified. E2F1 was overexpressed in most solid cancers including LIHC. E2F1 overexpression was correlated with poor prognosis in LIHC. Copy number alterations could positively affect E2F1 expression. Moreover, ceRNAs network was established with 3 long non-coding RNAs (lncRNAs) named AC025048.4, AC090114.2, and AC092171.5, as well as 4 microRNAs (miRNAs) including miR-150-5p, miR-302c-3p, miR-520d-3p, and miR-330-5p. Single cell sequencing data showed that E2F1 was mainly expressed in malignant cells and proliferating T cells, and that E2F targets almost exclusively enriched in proliferating T cells. Besides, there existed a positive correlation between E2F1 and certain immune cells including CD8(+) T cells, CD4(+) T cells, B cells, macrophages, and dendritic cells. Conclusions: This study elucidated that E2F1 could affect tumor development and immune microenvironment in LIHC. Thus, E2F1 might be a potential prognostic biomarker and therapeutic target for LIHC.

Evaluation of Hybrid VMAT Advantages and Robustness Considering Setup Errors Using Surface Guided Dose Accumulation for Internal Lymph Mammary Nodes Irradiation of Postmastectomy Radiotherapy.

Objectives: Setup error is a key factor affecting postmastectomy radiotherapy (PMRT) and irradiation of the internal mammary lymph nodes is the most investigated aspect for PMRT patients. In this study, we evaluated the robustness, radiobiological, and dosimetric benefits of the hybrid volumetric modulated arc therapy (H-VMAT) planning technique based on the setup error in dose accumulation using a surface-guided system for radiation therapy. Methods: We retrospectively selected 32 patients treated by a radiation oncologist and evaluated the clinical target volume (CTV), including internal lymph node irradiation (IMNIs), and considered the planning target volume (PTV) margin to be 5 mm. Three different planning techniques were evaluated: tangential-VMAT (T-VMAT), intensity-modulated radiation therapy (IMRT), and H-VMAT. The interfraction and intrafraction setup errors were analyzed in each field and the accumulated dose was evaluated as the patients underwent daily surface-guided monitoring. These parameters were included while evaluating CTV coverage, the dose required for the left anterior descending artery (LAD) and the left ventricle (LV), the normal tissue complication probability (NTCP) for the heart and lungs, and the second cancer complication probability (SCCP) for contralateral breast (CB). Results: When the setup error was accounted for dose accumulation, T-VMAT (95.51%) and H-VMAT (95.48%) had a higher CTV coverage than IMRT (91.25%). In the NTCP for the heart, H-VMAT (0.04%) was higher than T-VMAT (0.01%) and lower than IMRT (0.2%). However, the SCCP (1.05%) of CB using H-VMAT was lower than that using T-VMAT (2%) as well as delivery efficiency. And T-VMAT (3.72) and IMRT (10.5).had higher plan complexity than H-VMAT (3.71). Conclusions: In this study, based on the dose accumulation of setup error for patients with left-sided PMRT with IMNI, we found that the H-VMAT technique was superior for achieving an optimum balance between target coverage, OAR dose, complication probability, plan robustness, and complexity.

Low-Temperature Plasma-Activated Medium Inhibited Proliferation and Progression of Lung Cancer by Targeting the PI3K/Akt and MAPK Pathways.

Low-temperature plasma, an engineered technology to generate various reactive species, is actively studied in cancer treatment in recent years, yet mainly by using a traditional 2D cell culture system. In this study, we explored the effect of the plasma-activated medium (PAM) on lung cancer cells in vitro and in vivo by using a 3D cell culture model. The results showed that PAM markedly inhibited 3D spheroid formation and downregulated stemness-related gene expression. We found that reactive oxygen species (ROS) penetrated throughout the whole spheroids and induced cell death surrounding and in the core of the tumor spheroid. Besides, PAM treatment suppressed migration and invasion of lung cancer cells and downregulated epithelial-mesenchymal transition- (EMT-) related gene expression. In the mouse xenograft model, the tumor volume was significantly smaller in the PAM-treated group compared with the control group. By using transcriptome sequencing, we found that PI3K/Akt and MAPK pathways were involved in the inhibition effects of PAM on lung cancer cells. Therefore, our results indicated that PAM exhibits potential anticancer effects on lung cancer and provides insight into further exploration of PAM-induced cell death and translational preclinical use.

MicroRNA-15a promotes prostate cancer cell ferroptosis by inhibiting GPX4 expression.

Ferroptosis is a novel form of regulated cell death characterized by accumulated lipid reactive oxygen species (ROS) and inactivation of glutathione peroxidase 4 (GPX4). The present study aimed to investigate the role of microRNA (miRNA/miR)-15a in ferroptosis of prostate cancer cells. Bioinformatics analysis was performed to predict the potential interaction between miR-15a and the 3'-untranslated region (UTR) of GPX4 mRNA. The prostate cancer cell line, LNCAP was transfected with miR-15a mimics or small interfering (si)-GPX4. Reverse transcription-quantitative PCR and western blot analyses were performed to detect the mRNA and protein expression levels of GPX4, respectively. Biotin-RNA pull-down and dual-luciferase reporter assays were performed to verify the interaction between miR-15a and GPX4 mRNA. The Cell Counting Kit-8 assay was performed to assess cell proliferation, while lactate dehydrogenase (LDH) and intracellular ferrous iron levels were detected via ELISA. Lipid ROS and mitochondrial membrane potential (MMP) were assessed via flow cytometry and staining with C11-BIODIPY probes or JC-1. Furthermore, lipid peroxidation was identified by measuring malondialdehyde (MDA) levels. The results demonstrated that transfection with miR-15a mimics decreased GPX4 protein expression. Bioinformatics analysis revealed potential binding sites between miR-15a and the 3'-UTR region of GPX4, and RNA pull-down and the dual-luciferase reporter assays further confirmed the interaction between miR-15a and GPX4 mRNA. Both transfection with miR-15a mimics and si-GPX4 suppressed cell proliferation, elevated LDH release, accumulated intracellular ferrous iron and ROS, disrupted MMP and increased MDA levels. Taken together, the results of the present study suggest miR-15a induces ferroptosis by regulating GPX4 in prostate cancer cells, which provides evidence for investigating the therapeutic strategies of prostate cancer.

CHEK1: a hub gene related to poor prognosis for lung adenocarcinoma.

Aim: The study aims to pinpoint hub genes and investigate their functions in order to gain insightful understandings of lung adenocarcinoma (LUAD). Methods: Bioinformatic approaches were adopted to investigate genes in databases including Gene Expression Omnibus, WebGestalt, STRING and Cytoscape, GEPIA2, Oncomine, Human Protein Atlas, TIMER2.0, UALCAN, cBioPortal, TargetScanHuman, OncomiR, ENCORI, Kaplan-Meier plotter, UCSC Xena, European Molecular Biology Laboratory - European Bioinformatics Institute Single Cell Expression Atlas and CancerSEA. Results: Five hub genes were ascertained. CHEK1 was overexpressed in a range of cancers, including LUAD. Promoter methylation, amplification and miRNA regulation might trigger CHEK1 upregulation, signaling poor prognosis. CHEK1 with its coexpressed genes were enriched in the cell cycle pathway. Intratumor heterogeneity of CHEK1 expression could be observed. Cell clusters with CHEK1 expression were more prone to metastasis and epithelial-to-mesenchymal transition. Conclusion:CHEK1 might potentially act as a prognostic biomarker for LUAD.

Lay abstract Lung adenocarcinoma is the most common type of lung cancer. Many genes are thought to be linked to the development of cancers. We looked at key genes in lung adenocarcinoma by analyzing public databases. Five key genes, including CHEK1, were found. The amount of CHEK1 was higher in lung adenocarcinoma than in normal lung cells. The presence of a large amount of CHEK1 was linked to poor survival in people with lung adenocarcinoma. Also, CHEK1 was mainly expressed in some specific tumor cell clusters rather than in all tumor cell clusters. The clusters with CHEK1 expression were more cancerous compared with other clusters. Overall, CHEK1 might be a potential marker to predict survival in people with lung adenocarcinoma.

Upregulated long non­coding RNA LincIN promotes tumor progression via the regulation of nuclear factor 90/microRNA­7/HOXB13 in esophageal squamous cell carcinoma.

Long non­coding RNA LincIN has been reported to be overexpressed and to be involved in the metastasis of breast cancer. However, the expression and role of LincIN in esophageal squamous cell carcinoma (ESCC) remain unsolved. In the present study, LincIN expression was examined in ESCC by RT­qPCR, and the roles of LincIN in ESCC were determined using cell growth, migration and invasion assays. In addition, the effects of LincIN on nuclear factor 90 (NF90) and microRNA/miR (miR)­7 were examined by RNA immunoprecipitation assay, RT­qPCR, dual­luciferase reporter assay and western blot analysis. The results revealed that LincIN expression was significantly increased in ESCC tissues and cell lines. The increased expression of LincIN was positively associated with invasion depth, lymph node metastasis, TNM stage and a poor prognosis. Functional assays revealed that the overexpression of LincIN promoted ESCC cell growth, migration and invasion. Mechanistic analysis revealed that LincIN physically bound to NF90, enhanced the binding between NF90 and primary miR­7 (pri­miR­7), and further enhanced the inhibitory effects of NF90 on miR­7 biogenesis. Therefore, LincIN downregulated miR­7 expression in ESCC. The expression of miR­7 inversely correlated with that of LincIN in ESCC tissues. By downregulating miR­7, LincIN increased the expression of HOXB13, a target of miR­7. The overexpression of miR­7 or the depletion of HOXB13 both attenuated the tumor­promoting roles of LincIN in ESCC cell growth, migration and invasion. On the whole, the findings of the present study suggest that LincIN is overexpressed and plays an oncogenic role in ESCC via the regulation of the NF90/miR­7/HOXB13 axis. Thus, LincIN may prove to be a promising prognostic biomarker and therapeutic target for ESCC.

E2F7, regulated by miR­30c, inhibits apoptosis and promotes cell cycle of prostate cancer cells.

Prostate cancer (PCa) remains a leading cause of mortality among men in the United States and Western Europe. The molecular mechanism of PCa pathogenesis has not been fully elucidated. In the present study, the expression profile of E2F transcription factor 7 (E2F7) in PCa was examined using immunohistochemistry and reverse transcription­quantitative PCR, whilst cell cycle progression and apoptosis were determined using fluorescent cell activated sorting techniques. Cell viability was measured using Cell Counting Kit­8 in loss­ and gain­of­function studies. Dual­luciferase reporter assay was used to verify if E2F7 was one of the potential targets of miR­30c. The staining score of E2F7 of PCa tissues was found to be notably higher compared with that of adjacent normal tissues. Suppression of E2F7 expression in PCa cell lines led to significantly reduced proliferation rates, increased proportion of cells in the G1 phase of the cell cycle and higher apoptotic rates compared with those in negative control groups. Dual­luciferase reporter assay revealed E2F7 to be one of the binding targets of microRNA (miR)­30c. In addition, transfection of miR­30c mimics into PCa cells resulted in reduced cell viability, increased proportion of cells in the G1 phase and higher apoptotic rates. By contrast, transfection with the miR­30c inhibitor led to lower apoptosis rates of PCa cells compared with negative control groups, whilst E2F7 siRNA co­transfection reversed stimulatory effects of miR­30c inhibitors on cell viability. In addition, the expression of cyclin­dependent kinase inhibitor p21 were found to be upregulated by transfection with either E2F7 siRNA or miR­30c mimics into PCa cells. In conclusion, the present study suggested that E2F7 may be positively associated with PCa cell proliferation by inhibiting p21, whereas E2F7 is in turn under regulation by miR­30c. These observations suggest the miR­30c/E2F7/p21 axis to be a viable therapeutic target for PCa.

Expression of glucose transporter 1 and prognosis in non-small cell lung cancer: a pooled analysis of 1665 patients.

Glucose transporter 1 (GLUT1) plays an important role in the transport and metabolism of glucose in cancer cells. An increasing number of studies have explored the connection between GLUT1 expression and prognosis in non-small cell lung cancer (NSCLC), but the results have been controversial. Therefore, we conducted a meta-analysis to obtain a comprehensive evaluation of the prognostic value of GLUT1 in NSCLC. Relevant studies from PubMed, Embase, and Web of Science were searched. Hazard ratios (HRs) and odds ratios (ORs) with their 95% confidence intervals (CIs) were used as the effective measures. A total of 10 studies involving 1,665 patients were included in this meta-analysis. The results showed that GLUT1 overexpression was associated with poor overall survival (HR = 2.21; 95% CI, 1.42-3.42; p < 0.001) and disease-free survival (HR = 1.73; 95% CI, 1.35-2.23; p < 0.001). Furthermore, elevated GLUT1 expression correlated with sex (OR = 2.29; 95% CI, 1.17-4.49; p = 0.015), advanced tumor stage (OR = 2.46; 95% CI, 1.79-3.38; p < 0.001), histology (OR = 6.99; 95% CI, 4.71-10.38; p < 0.001), and large tumor size (OR = 2.77; 95% CI, 1.73-4.44; p < 0.001). This meta-analysis revealed overexpression of GLUT1 to be a biomarker of worse prognosis in NSCLC.

Preliminary comparison of the registration effect of 4D-CBCT and 3D-CBCT in image-guided radiotherapy of Stage IA non-small-cell lung cancer.

In this study, we compared the registration effectiveness of 4D cone-beam computed tomography (CBCT) and 3D-CBCT for image-guided radiotherapy in 20 Stage IA non-small-cell lung cancer (NSCLC) patients. Patients underwent 4D-CBCT and 3D-CBCT immediately before radiotherapy, and the X-ray Volume Imaging software system was used for image registration. We performed automatic bone registration and soft tissue registration between 4D-CBCT or 3D-CBCT and 4D-CT images; the regions of interest (ROIs) were the vertebral body on the layer corresponding to the tumor and the internal target volume region. The relative displacement of the gross tumor volume between the 4D-CBCT end-expiratory phase sequence and 4D-CT was used to evaluate the registration error. Among the 20 patients (12 males, 8 females; 35-67 years old; median age, 52 years), 3 had central NSCLC and 17 had peripheral NSCLC, 8 in the upper or middle lobe and 12 in the lower lobe (maximum tumor diameter range, 18-27 mm). The internal motion range in three-dimensional space was 12.52 ± 2.65 mm, accounting for 47.8 ± 15.3% of the maximum diameter of each tumor. The errors of image-guided registration using 4D-CBCT and 3D-CBCT on the x (left-right), y (superior-inferior), z (anterior-posterior) axes, and 3D space were 0.80 ± 0.21 mm and 1.08 ± 0.25 mm, 2.02 ± 0.46 mm and 3.30 ± 0.53 mm, 0.52 ± 0.16 mm and 0.85 ± 0.24 mm, and 2.25 ± 0.44 mm and 3.59 ± 0.48 mm (all P < 0.001), respectively. Thus, 4D-CBCT is preferable to 3D-CBCT for image guidance in small pulmonary tumors because 4D-CBCT can reduce the uncertainty in the tumor location resulting from internal motion caused by respiratory movements, thereby increasing the image-guidance accuracy.

Prognostic role of B7-H4 in patients with non-small cell lung cancer: A meta-analysis.

B7 homolog 4 (B7-H4) has been recently reported to be a prognostic marker in non-small cell lung cancer (NSCLC) in some studies. However, the results remained conflicting. Thus, we aimed to comprehensively assess the association between B7-H4 expression and prognosis of NSCLC patients by performing a meta-analysis. Relevant publications were thoroughly searched of PubMed, Embase, Web of Science and China National Knowledge Infrastructure (CNKI). The pooled odds ratios (ORs) and hazard ratios (HRs) with 95% confidence intervals (CIs) were applied to evaluate the effects. A total of 9 studies comprising 1444 patients were included in this meta-analysis. B7-H4 overexpression was associated with presence of lymph node metastasis (OR=3.59, 95%CI=2.39-5.38, p<0.001; fixed effect), advanced TNM stage (OR=2.36, 95%CI=1.2-4.67, p=0.013; random effect), and poor differentiation (OR=2.11, 95%CI=1.12-3.99, p=0.021; fixed effect). However, B7-H4 had no significant correlation with gender, age or histology in NSCLC. Furthermore, in a fixed effects model, the results indicated that B7-H4 overexpression was significantly associated with poor OS (HR=2.03, 95%CI=1.41-2.92, p<0.001). This meta-analysis demonstrated that high B7-H4 expression is an unfavorable prognostic factor in NSCLC. Because few studies were included for meta-analysis and almost all included studies were performed on Chinese patients, therefore; large scale prospective studies are needed to verify our results.

GSTP1 Ile105Val polymorphism and colorectal cancer risk: an updated analysis.

BACKGROUND: Many studies have investigated the association between the Glutathione S transferase-P1 (GSTP1) Ile105Val polymorphism and colorectal cancer (CRC) susceptibility, but the results were conflicting. The aim of this study is to quantitatively summarize the relationship between this polymorphism and CRC risk. METHODS: Two investigators independently searched the Medline, Embase, China National Knowledge Infrastructure (CNKI) and Chinese Biomedicine databases for studies published before December 2012. Summary odds ratios (ORs) and 95% confidence intervals (95% CIs) for GSTP1 polymorphism and CRC were calculated in a fixed-effects model (the Mantel-Haenszel method) and a random-effects model (the DerSimonian and Laird method) when appropriate. RESULTS: This meta-analysis included 29 case-control studies, which included 8160 CRC cases and 10,450 controls. Overall, the variant genotypes (ValVal and IleVal) of the Ile105Val were not associated with CRC risk when compared with the wild-type IleIle homozygote. Similarly, no associations were found in the dominant and recessive models. When stratifying for ethnicity, source of controls, study sample size and genotyping methods, no evidence of significant association was observed in any subgroup, except among those studies taking others as genotyping methods (recessive model, OR=0.71, 95%CI=0.52-0.96). Limiting the analysis to the studies within Hardy-Weinberg equilibrium, the results were persistent and robust. No publication bias was found in the present study. CONCLUSION: This updated meta-analysis suggests that the GSTP1 Ile105Val polymorphism may not be associated with CRC risk, while the observed decrease in risk of CRC may be due to small-study bias.