Objective:To investigate the effects of different doses of whole abdominal ultra‐high dose rate (FLASH) irradiation on the intestinal microbiota of mice.Methods:A total of 25 healthy male C57BL/6J mice were randomly divided into the control ( n=5) and FLASH irradiation groups ( n=20) by simple randomization method, and the FLASH irradiation group was further divided into different radiation dose subgroups of 10, 15, 20, 25 Gy, 5 in each group. The mice were irradiated with a single whole abdomen at a dose rate of 100 Gy/s, then sacrificed 3.5 d after irradiation. Fresh fecal specimens and intestinal tissues of mice were collected for 16S rRNA sequencing, microbiota analysis, hematoxylin eosin (HE) staining and injury severity score analysis. Two-group comparison was performed by independent sample t-test. Multi-group comparison was conducted by one-way ANOVA. Results:HE staining revealed that the whole abdomen FLASH irradiation caused varying degree of intestinal injury in mice, and the intestinal injury reaction was aggravated with the increase of irradiation dose. β‐diversity analyses showed that there were differences in the composition of intestinal microbiota between FLASH irradiation group and control group ( P=0.001), but the differences in the relative abundance of the species between the irradiation groups at different doses were relatively small, and there were their own dominant genera of bacteria. Comparison of different doses of FLASH irradiation groups with control group screened out 16 species of bacteria with shared differences at the genus level, in which Lactobacillus, Ligilactobacillus and unclassified Lactobacillus were more abundant in the control group, while Escherichia, Allobaculum, and Muribaculum were more abundant in the FLASH irradiation groups. Conclusions:The whole‐abdominal FLASH irradiation induces intestinal damage in mice, and the intestinal damage response is worsened with the increase of irradiation dose. Different doses of whole abdominal FLASH irradiation alter the intestinal microbiota composition of mice. Sixteen species of common intestinal differential microbiota at the genus level are screened out in the different doses of FLASH irradiation groups compared with the control group, which may serve as a marker for measuring intestinal injury in mice irradiated with whole‐abdominal FLASH.

Objective:To analyze the color Doppler flow imaging (CDFI) features of familial exudative vitreoretinopathy (FEVR) at different stages.Methods:A retrospective study. A total of 104 patients with 201 eyes from Department of Ophthalmology of Beijing Tongren Hospital who were hospitalized for fundus examination and diagnosed with FEVR from 2018 to 2022 were included. There were 69 male cases with 133 eyes and 35 female cases with 68 eyes. The age was ranged from 2 months to 11 years, with a mean age of 2.9 years. Fundus and CDFI examination were performed in both eyes. Fluorescein fundus angiography was performed in 72 cases (144 eyes). FEVR staging was conducted according to literature standards. The presence of avascular areas in the peripheral retina or abnormal retina neovascularization was stage 1; the presence of retinal neovascularization at the vitreoretinal interface in the avascular area was stage 2; partial retinal detachment without macula involvement was stage 3; partial retinal detachment involving the macula was stage 4; complete retinal detachment was stage 5. The CDFI ultrasound features of FEVR at different stages were analyzed. The CDFI image features of FEVR patients in different stages were observed.Results:Among the 104 patients, 97 (93.3%, 97/104) cases were binocular and 7 (6.7%, 7/104) cases were monocular. In 201 eyes, stages 1 to 5 of FEVR were 49 (24.4%, 49/201), 23 (11.4%, 23/201), 39 (19.4%, 39/201), 71 (35.3%, 71/201), and 19 (9.5%, 19/201) eyes, respectively. CDFI examination showed no abnormality or mild vitreous opacity in 49 eyes vitreous body at stage 1. Vitreous opacities were observed in all 23 eyes in stage 2, and the echo of the temporal ballwall was not smooth. In 39 eyes at stage 3, the anterior globular cluster echo in temporal peripheral eyes was observed in 17 eyes and partial retinal detachment was observed in 13 eyes. In 71 eyes at stage 4, 51 eyes had temporal or infratemporal retinal folds, and 20 eyes had temporal retinal detachment. All the 19 eyes in stage 5 had total retinal detachment, of which 15 eyes had closed "funnel-shaped" retinal detachment. Among the patients with retinal folds, 13 had bilateral folds, and the fellow eyes of the other 25 patients with unilateral folds all had vitreous opacity or clump echo in front of the temporal spherical wall. Blood flow signals could be detected on the retinal folds with Doppler imaging.Conclusions:The CDFI manifestations of FEVR patients at different stages have different characteristics. The possibility of FEVR should be considered when the temporal or infratemporal retinal folds of both eyes are present, as well as the retinal folds of one eye, the contralateral vitreous body opacity, or the anterior temporal peribulbar cluster echoes are present.

Objective:To apply mass spectrometry as well as bioinformatics techniques to analyze HBV peptides derived from Pol and X proteins presented by human immortalized B lymphocytes(BLCLs),and to screen for effective T-cell epitopes,which lays the foundation for the development of therapeutic vaccines.Methods:The group has constructed a genome-wide expression plasmid containing 1.2-fold HBVC2 isoforms and transfected it into BLCLs by electro-transfection,isolated and identified HBV peptides by LC-MS/MS,bioinformatically predicted peptide sequences in terms of sensitization,antigenicity,toxicity,HLA molecular affinity,and the ability of peptide HLA-class Ⅰ complex to bind to specific T cells,and retrieved reported sequences.Results:HBV peptides from lysates of five immortalized B cells(BLCLs-1 to BLCLs-5)carrying HBVC2 subtype-expressing recombinants were analyzed,and 141 peptides with sequence lengths of no less than 8 amino acid residues(≥8 aa)were successfully isolated and identified,of which 133 were derived from Pol,and 8 from X proteins.The 141 HBV peptides were analyzed for sensitization,antigenicity and toxicity,and 50 antigenic,non-toxic and sensitizing HBV peptides(47 from Pol and 3 from X protein)were screened for affinity analysis with HLA-class Ⅰ and Ⅱ molecules and for prediction of the binding ability of the peptide HLA-class Ⅰ complexes to specific T cells.Among these peptides,37 had affinity to the corresponding genotypes of HLA-class Ⅰ and Ⅱ molecules.Finally,we obtained 37 peptides with antigenic,nontoxic,and sensitizing properties with IC50<500 nmol/L to HLA-class Ⅰ and Ⅱ molecules,which have affinitied to the corresponding genotypes but have not been reported,and can be continued to be explored as potential epitopes.Finally,15 HBV peptide hotspot core regions were formed by intra-and inter-strain common,contained,overlapping or neighboring sequence analysis of 141 peptides,with 7 core region sequences with antigenic,nontoxic,and sensitizing properties restricted to the corresponding geno-types.Peptides with affinity IC50<500 nmol/L had an affinity core sequence overlap of no less than 8 amino acids,and can be priori-tized for candidate T cell epitopes for subsequent studies.Conclusion:Peptides derived from Pol and X proteins have been successfully isolated and identified,and potential T cell epitopes have been screened.

Objective:To apply mass spectrometry as well as bioinformatics techniques to analyze HBV peptides derived from Pol and X proteins presented by human immortalized B lymphocytes(BLCLs),and to screen for effective T-cell epitopes,which lays the foundation for the development of therapeutic vaccines.Methods:The group has constructed a genome-wide expression plasmid containing 1.2-fold HBVC2 isoforms and transfected it into BLCLs by electro-transfection,isolated and identified HBV peptides by LC-MS/MS,bioinformatically predicted peptide sequences in terms of sensitization,antigenicity,toxicity,HLA molecular affinity,and the ability of peptide HLA-class Ⅰ complex to bind to specific T cells,and retrieved reported sequences.Results:HBV peptides from lysates of five immortalized B cells(BLCLs-1 to BLCLs-5)carrying HBVC2 subtype-expressing recombinants were analyzed,and 141 peptides with sequence lengths of no less than 8 amino acid residues(≥8 aa)were successfully isolated and identified,of which 133 were derived from Pol,and 8 from X proteins.The 141 HBV peptides were analyzed for sensitization,antigenicity and toxicity,and 50 antigenic,non-toxic and sensitizing HBV peptides(47 from Pol and 3 from X protein)were screened for affinity analysis with HLA-class Ⅰ and Ⅱ molecules and for prediction of the binding ability of the peptide HLA-class Ⅰ complexes to specific T cells.Among these peptides,37 had affinity to the corresponding genotypes of HLA-class Ⅰ and Ⅱ molecules.Finally,we obtained 37 peptides with antigenic,nontoxic,and sensitizing properties with IC50<500 nmol/L to HLA-class Ⅰ and Ⅱ molecules,which have affinitied to the corresponding genotypes but have not been reported,and can be continued to be explored as potential epitopes.Finally,15 HBV peptide hotspot core regions were formed by intra-and inter-strain common,contained,overlapping or neighboring sequence analysis of 141 peptides,with 7 core region sequences with antigenic,nontoxic,and sensitizing properties restricted to the corresponding geno-types.Peptides with affinity IC50<500 nmol/L had an affinity core sequence overlap of no less than 8 amino acids,and can be priori-tized for candidate T cell epitopes for subsequent studies.Conclusion:Peptides derived from Pol and X proteins have been successfully isolated and identified,and potential T cell epitopes have been screened.

Objective:To analyze the color Doppler flow imaging (CDFI) features of familial exudative vitreoretinopathy (FEVR) at different stages.Methods:A retrospective study. A total of 104 patients with 201 eyes from Department of Ophthalmology of Beijing Tongren Hospital who were hospitalized for fundus examination and diagnosed with FEVR from 2018 to 2022 were included. There were 69 male cases with 133 eyes and 35 female cases with 68 eyes. The age was ranged from 2 months to 11 years, with a mean age of 2.9 years. Fundus and CDFI examination were performed in both eyes. Fluorescein fundus angiography was performed in 72 cases (144 eyes). FEVR staging was conducted according to literature standards. The presence of avascular areas in the peripheral retina or abnormal retina neovascularization was stage 1; the presence of retinal neovascularization at the vitreoretinal interface in the avascular area was stage 2; partial retinal detachment without macula involvement was stage 3; partial retinal detachment involving the macula was stage 4; complete retinal detachment was stage 5. The CDFI ultrasound features of FEVR at different stages were analyzed. The CDFI image features of FEVR patients in different stages were observed.Results:Among the 104 patients, 97 (93.3%, 97/104) cases were binocular and 7 (6.7%, 7/104) cases were monocular. In 201 eyes, stages 1 to 5 of FEVR were 49 (24.4%, 49/201), 23 (11.4%, 23/201), 39 (19.4%, 39/201), 71 (35.3%, 71/201), and 19 (9.5%, 19/201) eyes, respectively. CDFI examination showed no abnormality or mild vitreous opacity in 49 eyes vitreous body at stage 1. Vitreous opacities were observed in all 23 eyes in stage 2, and the echo of the temporal ballwall was not smooth. In 39 eyes at stage 3, the anterior globular cluster echo in temporal peripheral eyes was observed in 17 eyes and partial retinal detachment was observed in 13 eyes. In 71 eyes at stage 4, 51 eyes had temporal or infratemporal retinal folds, and 20 eyes had temporal retinal detachment. All the 19 eyes in stage 5 had total retinal detachment, of which 15 eyes had closed "funnel-shaped" retinal detachment. Among the patients with retinal folds, 13 had bilateral folds, and the fellow eyes of the other 25 patients with unilateral folds all had vitreous opacity or clump echo in front of the temporal spherical wall. Blood flow signals could be detected on the retinal folds with Doppler imaging.Conclusions:The CDFI manifestations of FEVR patients at different stages have different characteristics. The possibility of FEVR should be considered when the temporal or infratemporal retinal folds of both eyes are present, as well as the retinal folds of one eye, the contralateral vitreous body opacity, or the anterior temporal peribulbar cluster echoes are present.

Objective:To investigate the effects of different doses of whole abdominal ultra‐high dose rate (FLASH) irradiation on the intestinal microbiota of mice.Methods:A total of 25 healthy male C57BL/6J mice were randomly divided into the control ( n=5) and FLASH irradiation groups ( n=20) by simple randomization method, and the FLASH irradiation group was further divided into different radiation dose subgroups of 10, 15, 20, 25 Gy, 5 in each group. The mice were irradiated with a single whole abdomen at a dose rate of 100 Gy/s, then sacrificed 3.5 d after irradiation. Fresh fecal specimens and intestinal tissues of mice were collected for 16S rRNA sequencing, microbiota analysis, hematoxylin eosin (HE) staining and injury severity score analysis. Two-group comparison was performed by independent sample t-test. Multi-group comparison was conducted by one-way ANOVA. Results:HE staining revealed that the whole abdomen FLASH irradiation caused varying degree of intestinal injury in mice, and the intestinal injury reaction was aggravated with the increase of irradiation dose. β‐diversity analyses showed that there were differences in the composition of intestinal microbiota between FLASH irradiation group and control group ( P=0.001), but the differences in the relative abundance of the species between the irradiation groups at different doses were relatively small, and there were their own dominant genera of bacteria. Comparison of different doses of FLASH irradiation groups with control group screened out 16 species of bacteria with shared differences at the genus level, in which Lactobacillus, Ligilactobacillus and unclassified Lactobacillus were more abundant in the control group, while Escherichia, Allobaculum, and Muribaculum were more abundant in the FLASH irradiation groups. Conclusions:The whole‐abdominal FLASH irradiation induces intestinal damage in mice, and the intestinal damage response is worsened with the increase of irradiation dose. Different doses of whole abdominal FLASH irradiation alter the intestinal microbiota composition of mice. Sixteen species of common intestinal differential microbiota at the genus level are screened out in the different doses of FLASH irradiation groups compared with the control group, which may serve as a marker for measuring intestinal injury in mice irradiated with whole‐abdominal FLASH.

Objective:To investigate the effects of ultra-high dose rate radiation (FLASH-RT) and conventional radiation (CONV-RT) on plasma metabolites in glioma mice.Methods:Tocally 21 male C57BL/6J mice bearing intracranial glioma xenograft were randomly divided into healthy control group ( n=3), CONV-RT group ( n=9) and FLASH-RT group ( n=9). The CONV-RT group was administered a single dose of 24 Gy radiation on the head of mice at a dose rate of 0.4 Gy/s, and the FLASH-RT group was administered a single dose of 24 Gy radiation on the head of mice at a dose rate of 60 Gy/s, and the healthy control group was given 0 Gy pseudoradiation under the same condition. Mice blood was collected through the inner canthus vein for plasma separation at 1, 3 and 7 d after radiation in the two radiation groups, and the blood plasma of healthy control group was collected at 7 days after sham radiation. The changes in plasma metabolites were detected by the non-targeted metabolomics based on liquid chromatography mass spectrometry tandem platform. Results:After irradiation, the metabolites in plasma of two irradiation groups had significant difference. Compared with the healthy control group, 12 and 5 differential metabolites were screened out in the FLASH-RT group and CONV-RT group at three time points, respectively. The difference of plasma metabolites had the largest value at 1 day and decreased at 3 and 7 d after radiation. The arachidonic acid and isovaleric acid, involving arachidonic acid metabolism, biosynthesis of unsaturated fatty acids, and tyrosine metabolism pathways were screened in both FLASH-RT group and CONV-RT group, and the 10 differential metabolites, mainly involving energy metabolism and redox reactions, only existed in the FLASH-RT group.Conclusions:Arachidonic acid and isovaleric acid may be the common sensitive biomarkers to different radiation patterns, which provides ideas for further exploring the molecular mechanism of FLASH-RT in the treatment of glioma.

: AbstractObjective: To explore the effect and mechanism of curcumin on human T-cell acute lymphoblastic leukemia (T-ALL) cell apoptosis induced by Mcl-1 small molecule inhibitors UMI-77. METHODS: T-ALL cell line Molt-4 was cultured, and the cells were treated with different concentrations of curcumin and Mcl-1 small molecule inhibitor UMI-77 for 24 h. The MTT method was used to detect the cell survival rate after different treatment; According to the results of curcumin and UMI-77, the experimental settings were divided into control group, curcumin group (20 μmol/L curcumin treated cells), UMI-77 group (15 μmol/L Mcl-1 small molecule inhibitor UMI-77 treated cells) and curcumin+ UMI-77 group (20 μmol/L curcumin and 15 μmol/L Mcl-1 small molecule inhibitor UMI-77 treated cells), MTT method was used to detect cell proliferation inhibition rate, Annexin V-FITC/PI double staining method and TUNEL staining were used to detect cell apoptosis, DCFH-DA probe was used to detect cell reactive oxygen species, JC-1 fluorescent probe was used to detect mitochondrial membrane potential, Western blot was used to detect the expression levels of apoptosis-related proteins and Notch1 signaling pathway-related proteins. RESULTS: After the treatment of Molt-4 cells with different concentrations of curcumin and Mcl-1 small molecule inhibitor UMI-77, the cell survival rate was decreased (P<0.05); Compared with the control group, the cell proliferation inhibition rate of the curcumin group and the UMI-77 group were increased, the apoptosis rate of cell was increased, the level of ROS was increased, the protein expression of Bax, Caspase-3 and Caspase-9 in the cells were all increased, and the protein expression of Bcl-2 was reduced (P<0.05); Compared with the curcumin group or UMI-77 group, the cell proliferation inhibition rate and apoptosis rate of the curcumin+UMI-77 group were further increased, and the level of ROS was increased. At the same time, the protein expression of Bax, Caspase-3 and Caspase-9 in the cells were all increased, the protein expression of Bcl-2 was reduced (P<0.05); In addition, the mitochondrial membrane potential of the cells after curcumin treatment was decreased, and the proteins expression of Notch1 and HES1 were reduced (P<0.05). CONCLUSION Curcumin can enhance the apoptosis of T-ALL cells induced by Mcl-1 small molecule inhibitor UMI-77 by reducing the mitochondrial membrane potential, the mechanism may be related to the inhibition of Notch1 signaling pathway.
Apoptosis ; Apoptosis Regulatory Proteins ; Caspase 3/metabolism* ; Caspase 9/pharmacology* ; Cell Line, Tumor ; Curcumin/pharmacology* ; Humans ; Myeloid Cell Leukemia Sequence 1 Protein/metabolism* ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; Proto-Oncogene Proteins c-bcl-2/metabolism* ; Reactive Oxygen Species/pharmacology* ; Sulfonamides ; Thioglycolates ; bcl-2-Associated X Protein/pharmacology*

Objective: To analyze and compare the distribution of the high-risk population of upper gastrointestinal (UGI) cancer and the factors influencing the compliance rate of endoscopic screening in urban China and rural China. Methods: From 2015 to 2017, an epidemiological survey was conducted on residents aged 40-69 in two rural areas (Luoshan county of Henan province, Sheyang county of Jiangsu province) and two urban areas (Changsha city of Hunan province, Harbin city of Heilongjiang province). As a result, high-risk individuals were recommended for endoscopic screening. Chi-square χ(2) test was used to compare the high-risk rate of UGI cancer between urban and rural residents. In addition, the multivariate logistic regression model was used to analyze the factors influencing the compliance rate of endoscopic screening. Results: A total of 48, 310 residents aged 40-69 were enrolled in this study, including 22 870 (47.34%) residents from rural areas and 25 440 (52.66%) residents from urban areas. A total of 23 532 individuals were assessed with a high risk of UGI cancer, with an overall risk rate of 48.71%. A higher proportion of participants with high risk was observed in rural China (56.17%, 12 845/22 870) than in urban China (42.01%, 10 687/25 440). A total of 10 971 high-risk individuals with UGI cancer participated in endoscopic screening, with an overall compliance rate of 46.62% (10 971/23 532), 45.15% (5 799/12 845) in rural China, and 48.40% (5 172/10 687) in urban China. In rural population, the compliance rate of endoscopic screening was higher in those of females, aged 50-69 years, primary school education or above, high income, a family history of UGI cancer, history of gastric and duodenal ulcer, history of reflux esophagitis, and history of superficial gastritis, but lower in smokers (P<0.05). Among the urban population, the compliance rate of endoscopic screening was higher in those aged 40-49 years, uneducated, low income, family history of UGI cancer, history of reflux esophagitis, history of superficial gastritis, but lower in smokers (P<0.05). Conclusions: The proportion of participants with high risk of UGI cancer in rural areas is higher than that of urban areas. The compliance rates of endoscopic screening in urban and rural areas are low, and influencing factors of endoscopic screening exhibit some differences in rural China and urban China.
China/epidemiology* ; Early Detection of Cancer ; Esophagitis, Peptic ; Female ; Gastritis ; Gastrointestinal Neoplasms/epidemiology* ; Humans ; Rural Population ; Urban Population

Objective:To investigate the value of ADC derived from DWI combined with texture analysis derived from T 2WI fat suppressed images in distinguishing benign and malignant soft tissue tumors. Methods:The MRI and DWI images of 94 patients with soft tissue tumors (44 cases with malignant and 50 cases with benign) confirmed by pathology were analyzed retrospectively in the First Affiliated Hospital of USTC West District. ADC values of solid components were measured at GE ADW4.6 workstation. The texture features were extracted by manually drawing the ROI on the maximum level of the T 2WI fat suppressed images; the ADC values and texture parameters between the two groups were statistically analyzed by SPSS17.0, and the multivariate logistic regression model were conducted to analyze and calculate the diagnostic performance. Results:ADC value of benign and malignant soft tissue tumors was (1.6±0.3)×10 -3 mm 2/s, (1.2±0.5)×10 -3 mm 2/s, respectively, and the difference was statistically significant( t=-5.382, P<0.05). Taking 1.28×10 -3 mm 2/s as the critical value, the area under curve (AUC) for the diagnosis of benign and malignant soft tissue tumors was 0.783, the sensitivity was 92.00%, and the specificity was 65.91%. Among the texture features, the AUC of frequency size, skewness, Inertia All Direction_offset7, Inverse Difference Moment angle0_offset1, Inverse Difference Moment angle0_offset7 and Haralick Correlation All Direction_offset4_SD distinguishing benign and malignant soft tissue tumors were 0.825, 0.739, 0.826, 0.816, 0.820 and 0.783, respectively. The AUC, sensitivity and specificity of the best predictive model distinguishing benign and malignant soft tissue tumors were 0.930, 88.00% and 86.36% respectively using multivariate logistic regression analysis. Conclusion:ADC combined with texture analysis is of great value in preoperative differentiation of benign and malignant soft tissue tumors.