OBJECTIVES: To explore the expression profile of circular RNAs (circRNAs) and their potential roles in prognosis and progression of Wilms' tumor (WT). METHODS: Four pairs of WT and adjacent tissues were collected for high-throughput circRNA sequencing to identify the differentially expressed circular RNAs. RT-qPCR was used to verify the expression levels of the top 6 candidate circRNAs in the clinical samples. hsa_circ_0001900 was selected for analysis of its correlation with clinicopathological features and prognosis in 34 patients with WT. Sanger sequencing and RNase R digestion experiments were used to verify the cycling site and structural stability of hsa_circ_0001900 molecule. RESULTS: A total of 23 978 circular RNA molecules were identified in WT tissues by high-throughput circular RNA sequencing, and among them 614 were differentially expressed in WT. hsa_circ_0001900 showed the highest expression level among the differentially expressed circRNAs, which was consistent with the findings in clinical tumor samples and the sequencing results. Correlation analysis showed that hsa_circ_0001900 expression level was positively correlated with WT volume, and the children with high hsa_circ_0001900 expression had a lowered recurrence-free survival rate. The results of Sanger sequencing verified the circular splice site sequence of the molecule, and Rnase R digestion assay confirmed its stable covalent structure. CONCLUSIONS This study presents a comprehensive expression profile of circular RNAs in WT, and the expression level of hsa_circ_0001900 is related to the size of WT and the patients' prognosis, suggesting its possible role as a key driving gene in WT progression.
Humans ; RNA, Circular ; Wilms Tumor/pathology* ; Prognosis ; High-Throughput Nucleotide Sequencing ; Kidney Neoplasms/genetics* ; Sequence Analysis, RNA ; Male ; Female

Objective To identify the key genes differentially expressed in Wilms tumor and analyze their potential impacts on prognosis and immune responses of the patients. Methods High-throughput RNA sequencing was used to identify the differentially expressed mRNAs in clinical samples of Wilms tumor and paired normal tissues, and their biological functions were analyzed using GO, KEGG and GSEA enrichment analyses. The hub genes were identified using STRING database, based on which a prognostic model was constructed using LASSO regression. The mutations of the key hub genes were analyzed and their impacts on immunotherapy efficacy was predicted using the cBioPortal platform. RT-qPCR was used to verify the differential expressions of the key hub genes in Wilms tumor. Results Of the 1612 differentially expressed genes identified in Wilms tumor, 1030 were up-regulated and 582 were down-regulated, involving mainly cell cycle processes and immune responses. Ten hub genes were identified, among which 4 genes (TP53, MED1, CCNB1 and EGF) were closely related to the survival of children with Wilms tumor. A 3-gene prognostic signature was constructed through LASSO regression analysis, and the patients stratified into with high- and low-risk groups based on this signature had significantly different survival outcomes (HR=1.814, log-rank P=0.002). The AUCs of the 3-, 5-and 7-year survival ROC curves of this model were all greater than 0.7. The overall mutations in the key hub genes or the individual mutations in TP53/CCNB1 were strongly correlated with a lower survival rates, and a high TP53 expression was correlated with a poor immunotherapy efficacy. RT-qPCR confirmed that the key hub genes had significant differential expressions in Wilms tumor tissues and cells. Conclusion TP53 gene plays an important role in the Wilms tumor and may potentially serve as a new immunotherapeutic biomarker as well as a therapeutic target.

Objective To identify the key genes differentially expressed in Wilms tumor and analyze their potential impacts on prognosis and immune responses of the patients. Methods High-throughput RNA sequencing was used to identify the differentially expressed mRNAs in clinical samples of Wilms tumor and paired normal tissues, and their biological functions were analyzed using GO, KEGG and GSEA enrichment analyses. The hub genes were identified using STRING database, based on which a prognostic model was constructed using LASSO regression. The mutations of the key hub genes were analyzed and their impacts on immunotherapy efficacy was predicted using the cBioPortal platform. RT-qPCR was used to verify the differential expressions of the key hub genes in Wilms tumor. Results Of the 1612 differentially expressed genes identified in Wilms tumor, 1030 were up-regulated and 582 were down-regulated, involving mainly cell cycle processes and immune responses. Ten hub genes were identified, among which 4 genes (TP53, MED1, CCNB1 and EGF) were closely related to the survival of children with Wilms tumor. A 3-gene prognostic signature was constructed through LASSO regression analysis, and the patients stratified into with high- and low-risk groups based on this signature had significantly different survival outcomes (HR=1.814, log-rank P=0.002). The AUCs of the 3-, 5-and 7-year survival ROC curves of this model were all greater than 0.7. The overall mutations in the key hub genes or the individual mutations in TP53/CCNB1 were strongly correlated with a lower survival rates, and a high TP53 expression was correlated with a poor immunotherapy efficacy. RT-qPCR confirmed that the key hub genes had significant differential expressions in Wilms tumor tissues and cells. Conclusion TP53 gene plays an important role in the Wilms tumor and may potentially serve as a new immunotherapeutic biomarker as well as a therapeutic target.

Objective:To evaluate the effects of whole brain irradiation (WBI) and fecal microbiota transplantation (FMT) on hippocampal neurogenesis and the composition of gut microbiota in mice.Methods:Forty specific pathogen free ICR male mice (8-week-old, weighed 30 g) were divided into four groups by simple random sample method: control group (group C), radiation group (group R), group C+FMT and group R+FMT, 10 in each group. Animal models were established by WBI at a dose of 10 Gy by 4 MeV electron beam. In group C+FMT and group R+FMT, mice were gavaged with normal fecal bacteria suspension on day 2 post-irradiation, while those in group C and group R were gavaged with phosphate buffered saline as alternative. Hippocampal tissues and feces in four groups were collected on day 15 post-irradiation. 16S rRNA sequencing was used to detect the species and abundance of fecal flora. BrdU +/NeuN + immunofluorescence staining was performed to observe the neurogenesis in hippocampus of mice. Results:WBI and FMT had no effect on survival rate and body weight of mice. WBI induced the inhibition of hippocampal neurogenesis and flora disorder. The quantity of Bacteroideae and Rumen bacteria was increased by 28.6% and 102.9%, whereas that of Lactobacillus was significantly decreased by 70.6% ( P<0.05). FMT regulated the abundance of bacteria. The abundance of Enterobacteriaceae was significantly declined by 65.1% ( P=0.028), while that of Lactobacillus was increased by 58.2% ( P=0.015). FMT also promoted hippocampal neurogenesis to some extent after WBI. Conclusions:This preliminary study demonstrates that FMT alleviates the inhibition of hippocampal neurogenesis and flora disorder induced by WBI in mice. Ionizing radiation directly acting on the whole brain of mice indirectly disturbs the composition of gut microbiota, which in turn affects the degree of hippocampal neurogenesis in the brain of mice. There is a bidirectional interaction between gut microbiota and brain.

Objective:To explore the effect of cognitive behavioral intervention combined with early functional exercise on kinesiophobia in patients with lumbar disc herniation after surgery.Methods:From September 2020 to September 2021, the convenient sampling was used to select 90 patients with kinesiophobia after lumbar disc herniation surgery in Department of Orthopedics in Nanjing First Hospital as the research objects. They were divided into the intervention group and the control group by the random number table, with 45 cases in each group. The control group received routine nursing, while the intervention group received cognitive behavioral intervention combined with early functional exercise on the basis of routine nursing. The scores of Tampa Scale for Kinesiophobia (TSK) , Fear Avoidance Beliefs Questionnaire (FABQ) , Exercise Adherence Rating Scale (EARS) and Japanese Orthopaedic Association (JOA) were compared between the two groups at the time of admission, at discharge and 1 and 3 months after the operation.Results:After the intervention, the TSK and FABQ scores of the patients in the intervention group were lower than those in the control group at the time of discharge, 1 month after surgery and 3 months after surgery; the EARS and JOA scores of patients in the intervention group were higher than those in the control group at the time of discharge, 1 month and 3 months after operation, and the differences were all statistically significant ( P<0.05) . Conclusions:Cognitive-behavioral intervention combined with early functional exercise can effectively reduce the level of kinesiophobia and fear-avoidance beliefs in patients with postoperative lumbar disc herniation, improve their exercise compliance and effectively improve their physical function.

OBJECTIVES: To study the feasibility of ArcCHECK-3DVH system in dosimetric verification for stereotactic body radiaotherapy (SBRT) with flattening filter free (FFF) model. METHODS: SBRT treatment plans for 57 patients were introduced into ArcCHECK phantom and recalculated. The calculated dose distribution of treatment planning system and the measured dose distribution of ArcCHECK phantom were compared by γ analysis. Then the 3 dimensional dose distribution of target and organs at risk was reconstructed by 3DVH software. The reconstructed dose and calculated dose with treatment planning system (TPS) were compared, and the dose volume γ pass rate and deviation of dose volume parameters to the target and organs at risk were quantitatively valuated. RESULTS: Based on the threshold criteria (3%, 3 mm, 10%), namely the deviation of measuring points between the planned value and the measured value was less than 3%, and the proportion of points with similar values in the plane or sphere with the center of the point and the radius of 3 mm was 10%, the relative and absolute dose pass rates of SBRT treatment plans in ArcCHECK system via γ analysis were greater than 95%. Based on the stricter threshold criteria (2%, 2 mm, 10%), the relative and absolute dose pass rates of SBRT treatment plan in ArcCHECK system via γ analysis were about 93%. In 3DVH dose verification, the γ pass rate of target and organs at risk was exceed 97%, and the deviations in 3DVH of the target and organs at risk were less than ±5%. CONCLUSIONS The ArcCHECK-3DVH system in dose verification can provide more comprehensive dose distribution information to reasonably evaluate the SBRT plan, with more significance for guiding clinical treatment.
Humans ; Phantoms, Imaging ; Quality Assurance, Health Care ; Radiometry ; Radiosurgery ; Radiotherapy Dosage ; Radiotherapy Planning, Computer-Assisted ; Radiotherapy, Intensity-Modulated

Recently,the relationship between intestinal flora and its metabolites and tumorigenesis,inflammatory bowel diseasesas well as radiation-induced intestinal injury has captivated widespread attention from researchers.Accumulated evidence derived from nuclear accident investigation,animal model experiment and clinical research has proven the role of intestinal flora and its metabolites as the biomarkers to evaluate the radiation dose and severity of radiation-induced intestinal injury.This article reviews the relationship between intestinal flora and its metabolites and radiation-induced intestinal injury,aiming to provide theoretical reference for assessing the risk of radiation-induced intestinal injury.

Objective Toexplorethecorrelationanalysisofmammographyandclinicalcharacteristicsassociatedwiththerecurrence ofearlystageoftripleGnegativebreastcancer(TNBC).Methods Thedataofmammographyandclinicalcharacteristicsof231TNBC patientsfromJanuary2009to October2017 wereretrospectivelyanalyzed,andthenthedifferencesofmammographyandclinical characteristicswerecomparedbetweenthepatientswithrecurrence(n＝64)andthosewithoutrecurrence(n＝167).Results Compared withthepatientswithoutrecurrence,thepresenceoffamilyhistoryinpatientswithrecurrencewashigherthanthosewithoutrecurrence (12.5%vs3.6%,P＜0.05).Thehistologicalgradesofthepatientswithrecurrenceweremainlyshowedasgrade2(6.3%)and3(93.7%), andthehistologicalgradeinpatientswithrecurrencewashigherthanthosewithoutrecurrence(P＜0.05).Themammographicfindingsin TNBCshowedthatthepatientswithrecurrenceweremorelikelytohavedensebreasttissue(84.4%),axillarylymphnodesinvasion (53.1%)andlowercalcification(12.5%)(P＜0.05).Multivariatelogisticregressionanalysisshowedthatthefamilyhistoryofbreast cancer(P＝0.01,OR＝2.54),histologicalgradeof2(P＝0.01,OR＝2.23)and3(P＜0.001,OR＝3.79),mammographicdensity breasttissue(P＝0.02,OR＝2.32),calcification(P＝0.02,OR＝0.45),andaxillarylymphnodesinvasion(P＝0.03,OR＝1.75)were statisticallysignificancewithTNBCrecurrence.Conclusion Thefamilyhistory,histologicalgrade,thepresenceofdensebreasttissue andaxillarylymphnodesinvasionsatmammographywereassociatedwiththeincreasedriskofrecurrenceinTNBC,whilethepresenceof calcificationwasrelatedtothelowerriskofrecurrenceinTNBC.

Understanding the effect of immunosuppressive agents on intestinal microbiota is important to reduce the mortality and morbidity from orthotopic liver transplantation (OLT). We investigated the relationship between the commonly used immunosuppressive agent cyclosporine A (CSA) and the intestinal microbial variation in an OLT model. The rat samples were divided as follows: (1) N group (normal control); (2) I group (isograft LT, Brown Norway [BN] rat to BN); (3) R group (allograft LT, Lewis to BN rat); and (4) CSA group (R group treated with CSA). The intestinal microbiota was assayed by denaturing gradient gel electrophoresis profiles and by using real-time polymerase chain reaction. The liver histopathology and the alanine/aspartate aminotransferase ratio after LT were both ameliorated by CSA. In the CSA group, the numbers of rDNA gene copies of Clostridium cluster I, Clostridium cluster XIV, and Enterobacteriaceae decreased, whereas those of Faecalibacterium prausnitzii increased compared with the R group. Cluster analysis indicated that the samples from the N, I, and CSA groups were clustered, whereas the other clusters contained the samples from the R group. Hence, CSA ameliorates hepatic graft injury and partially restores gut microbiota following LT, and these may benefit hepatic graft rejection.

Objective To investigate the function of JAK2-STAT3 signaling pathway in pancreatic injury and systematic inflammatory response in rats with acute necrotizing pancreatitis ( ANP) . Methods SD rats were randomly divided into the ANP group (n=48), ANP+JAK2 inhibitor Ruxolitinib group (ANP+R group, n=48), ANP+STAT3 inhibitot Stattic group (ANP+S group, n=48), ANP+Ruxolitinib+Stattic group (ANP+R+S group, n=48), and sham operation group (SO group, n=48). 5％ sodium taurocholate injection via retrograde pancreatobiliary duct was used to establish ANP model. Blood samples from abdominal aorta and pancreatic tissue were collected after 3 h, 6 h, 12 h and 18 h after modeling. Serum amylase (AMY) and tumor necrosis factor-α(TNF-α) and interleukin-4 (IL-4) were tested. JAK2 and STAT3 mRNA expression and protein expression of p-JAK2 and p-STAT3 in pancreas were examined by RT qPCR and western blot, respectively. Results AMY, TNF-α and IL-4 in plasma, and JAK2 mRNA, STAT3 mRNA, p-JAK2 protein and p-STAT3 protein at different time points in ANP group were all obviously higher than those in SO group(P<0. 05). Serum AMY, TNF-αand IL-4 in ANP+R group, ANP+S group and ANP+R+S group at different time points were lower than those in ANP group [12 h (5391 ± 1009),(6130 ± 1227),(4818 ± 992)U/L vs (8524 ± 1360) U/L;(147.25 ± 27.85),(156.25 ± 23.17),(127.87 ± 21.39) ng/L vs (187.58 ±20.09)ng/L;(45.89 ±16.95),(50.19 ±15.87),(38.87 ±14.03)ng/L vs (58.85 ±9.34)ng/L] . JAK2 mRNA and p-JAK2 protein,STAT3 mRNA and p-STAT3 protein in ANP+R group and ANP+R+S group at different time points were obviously lower than those in ANP group (12 h 0. 357 ± 0. 091 vs 0. 597 ± 0. 121,1. 115 ± 0. 203 vs 1. 217 ± 0. 213,0. 361 ± 0. 089 vs 0. 489 ± 0. 097,0. 965 ± 0. 189 vs 1. 128 ± 0. 217, 0. 362 ± 0. 092 vs 0. 597 ± 0. 121,1. 107 ± 0. 212 vs 1. 217 ± 0. 213,0. 297 ± 0. 087 vs 0. 489 ± 0. 097,0. 713 ± 0. 184 vs 1. 128 ± 0. 217). STAT3 mRNA and p-STAT3 protein in ANP+S group were obviously lower than those in ANP group(0. 319 ± 0. 107 vs 0. 489 ± 0. 097,0. 849 ± 0. 177 vs 1. 128 ± 0. 217), and the difference was statistically different (P<0.05). Conclusions The activation of JAK2-STAT3 signaling pathway in pancreas may play a key role in the pathogenesis of systematic inflammatory response in ANP.