AIM: To detect the distribution and expression of vascular endothelial growth factor(VEGF)in radiation retinopathy(RR)through fluorescence targeted imaging.METHODS:Covalent binding of fluorescein FITC with VEGF antibody ranibizumab to prepare targeted fluorescent imaging probe ranibizumab-FITC. SD rats were randomly divided into three groups based on the principle of weight balance: a normal control group(Con group), a low-dose radiation group(10 Gy group), and a high-dose radiation group(30 Gy group). Medical linear accelerators and lead blocks were used to locally irradiate the rat eyeballs for modeling. Western blot and qRT-PCR were used to detect the expression levels of VEGF-A in each group and to screen for appropriate modeling dose. The inverted fluorescence microscope and the confocal microscope were used to observe the distribution of VEGF and imaging probes in the retinas of control and RR model group rats, and to verify the effectiveness of targeted probes.RESULTS:The expression level of VEGF-A in the retina of rats in the high-dose radiation group(30 Gy group)was higher than that in the normal control group(Con group). In early RR, VEGF expression was observed to be associated with microaneurysms and abnormal microvessels in the retina. VEGF accumulation was observed at the site of capillary wall damage. When retinal capillary endothelial damage occurred, targeted probes gathered on the outer surface of the vessel wall.CONCLUSION:The expression level of VEGF in the retina of RR model rats is elevated, and fluorescent targeted molecular imaging probes can detect the spatial distribution of VEGF at the microvascular lesions in the retina of RR rats.

Objective To construct large medical model named by "Huaxi HongYi"and explore its application effectiveness in assisting medical record generation. Methods By the way of a full-chain medical large model construction paradigm of "data annotation - model training - scenario incubation", through strategies such as multimodal data fusion, domain adaptation training, and localization of hardware adaptation, "Huaxi HongYi" with 72 billion parameters was constructed. Combined with technologies such as speech recognition, knowledge graphs, and reinforcement learning, an application system for assisting in the generation of medical records was developed. Results Taking the assisted generation of discharge records as an example, in the pilot department, after using the application system, the average completion times of writing a medical records shortened (21 min vs. 5 min) with efficiency increased by 3.2 time, the accuracy rate of the model output reached 92.4%. Conclusion It is feasible for medical institutions to build independently controllable medical large models and incubate various applications based on these models, providing a reference pathway for artificial intelligence development in similar institutions.

Objective To explore the prognostic value and immune effects of insulin-like growth factor family 2(IGFL2)in bladder cancer based on data from TCGA,TIMER2.0 and HPA.Methods Bioinformatics analysis was conducted to analyze the expression of IGFL2 in bladder cancer and its relationship with survival and prognosis of bladder cancer patients.The receiver operating characteristic(ROC)curve and prognostic nomogram were plotted to analyze the diagnostic value of IGFL2 for bladder cancer and its ability to predict the 5-year overall survival rate of patients.IGFL2-related signaling pathways were analyzed with GSEA enrichment.The correlation between IGFL2 expression and immune cells was analyzed based on data from the TIMER database.Results IGFL2 was highly expressed in bladder cancer(P＜0.001).The high expression of IGFL2 was positively correlated with patients'age,pathological stage,histological grade and T stage(all P＜0.05).Patients'with high IGFL2 expression had shorter overall survival and disease-specific survival than those with low IGFL2 expression(all P＜0.05).IGFL2 could be used as an independent prognostic factor for bladder cancer with good diagnostic and predictive value.GSEA enrichment analysis showed that IGFL2 was involved in signaling pathways related to immunity,inflammation and tumor.High IGFL2 expression was positively correlated with the infiltration of most immune cells.Conclusion IGFL2 is highly expressed in bladder cancer and can be a biomarker for the diagnosis and prognosis of this disease.It may become a potential target for immunotherapy.

Sennoside A (SA), a typical prodrug, exerts its laxative effect only after its transformation into rheinanthrone catalyzed by gut microbial hydrolases and reductases. Hydrolases have been identified, but reductases remain unknown. By linking a photoreactive group to the SA scaffold, we synthesized a photoaffinity probe to covalently label SA reductases and identified SA reductases using activity-based protein profiling (ABPP). From lysates of an active strain, Bifidobacterium pseudocatenulatum (B. pseudocatenulatum), 397 proteins were enriched and subsequently identified using mass spectrometry (MS). Among these proteins, chromate reductase/nicotinamide adenine dinucleotide (NADH) phosphate (NADPH)-dependent flavin mononucleotide (FMN) reductase/oxygen-insensitive NADPH nitroreductase (nfrA) was identified as a potent SA reductase through further bioinformatic analysis and The Universal Protein Resource (UniProt) database screening. We also determined that recombinant nfrA could reduce SA. Our study contributes to further illuminating mechanisms of SA transformation to rheinanthrone and simultaneously offers an effective method to identify gut bacterial reductases.

Objective:To investigate the effects of 177Lu-prostate specific membrane antigen (PSMA)-I&T combined with poly (adenosine diphosphate-ribose) polymerase (PARP) inhibitor (PARPi) fluzoparib on the proliferation and migration of prostate cancer cells and the tumor inhibitory effects. Methods:177Lu-PSMA-I&T was synthesized. Cytotoxicity assay, colony formation assay, 5-ethynyl-2′-deoxyuridine (EdU) cell proliferation assay, Transwell cell migration assay, and terminal-deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay, flow cytometry were performed to detect apoptosis and cell cycles. 22RV1 tumor-bearing mice models ( n=16) were established, and were randomly divided into 4 groups: control group (no treatment; n=4), fluzoparib monotherapy group (6mg/kg; n=4), 177Lu-PSMA-I&T monotherapy group (14.8MBq; n=4) and combination group (14.8MBq 177Lu-PSMA-I&T+ 6mg/kg fluzoparib; n=4). All mice were treated for 14 d. Tumor volume and body mass changes of tumor-bearing mice were observed and recorded. After the treatment, 18F-FDG PET/CT was performed to evaluate the tumor′s uptake of 18F-FDG. Effects of 177Lu-PSMA-I&T combined with fluzoparib on cell and tumor-bearing mice were observed. One-way analysis of variance and the least significant difference t test were used to analyze the data. Results:At half maximal inhibitory concentrations (IC 50) of 177Lu-PSMA-I&T (13.06MBq/ml) and fluzoparib (72.13μmol/L), compared to the fluzoparib monotherapy group and the 177Lu-PSMA-I&T monotherapy group, the combination treatment significantly enhanced the anti-tumor effect on 22RV1 cells, inhibited the DNA synthesis rate and colony-forming ability of 22RV1 cells, reduced cell migration rate, increased the percentage of DNA damage, resulted in a higher proportion of cells arrested in the G2/M phase and increased the apoptosis rate ( F values: 9.77-162.20, t values: 2.98-21.60, all P<0.05). Compared to the fluzoparib monotherapy group and the 177Lu-PSMA-I&T monotherapy group, the combination treatment resulted in a significant reduction in relative tumor volume (RTV%) 14 d post-administration and markedly decreased 18F-FDG uptake ( F values: 25.28 and 67.42, t values: 4.64-8.61, P values: 0.001-0.009). Conclusion:The combination of 177Lu-PSMA-I&T and fluzoparib can inhibit prostate cancer cell proliferation and migration, suppress tumor growth and metabolism, and demonstrates synergistic effects more effectively.

Objective:To investigate the effects of 177Lu-prostate specific membrane antigen (PSMA)-I&T combined with poly (adenosine diphosphate-ribose) polymerase (PARP) inhibitor (PARPi) fluzoparib on the proliferation and migration of prostate cancer cells and the tumor inhibitory effects. Methods:177Lu-PSMA-I&T was synthesized. Cytotoxicity assay, colony formation assay, 5-ethynyl-2′-deoxyuridine (EdU) cell proliferation assay, Transwell cell migration assay, and terminal-deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay, flow cytometry were performed to detect apoptosis and cell cycles. 22RV1 tumor-bearing mice models ( n=16) were established, and were randomly divided into 4 groups: control group (no treatment; n=4), fluzoparib monotherapy group (6mg/kg; n=4), 177Lu-PSMA-I&T monotherapy group (14.8MBq; n=4) and combination group (14.8MBq 177Lu-PSMA-I&T+ 6mg/kg fluzoparib; n=4). All mice were treated for 14 d. Tumor volume and body mass changes of tumor-bearing mice were observed and recorded. After the treatment, 18F-FDG PET/CT was performed to evaluate the tumor′s uptake of 18F-FDG. Effects of 177Lu-PSMA-I&T combined with fluzoparib on cell and tumor-bearing mice were observed. One-way analysis of variance and the least significant difference t test were used to analyze the data. Results:At half maximal inhibitory concentrations (IC 50) of 177Lu-PSMA-I&T (13.06MBq/ml) and fluzoparib (72.13μmol/L), compared to the fluzoparib monotherapy group and the 177Lu-PSMA-I&T monotherapy group, the combination treatment significantly enhanced the anti-tumor effect on 22RV1 cells, inhibited the DNA synthesis rate and colony-forming ability of 22RV1 cells, reduced cell migration rate, increased the percentage of DNA damage, resulted in a higher proportion of cells arrested in the G2/M phase and increased the apoptosis rate ( F values: 9.77-162.20, t values: 2.98-21.60, all P<0.05). Compared to the fluzoparib monotherapy group and the 177Lu-PSMA-I&T monotherapy group, the combination treatment resulted in a significant reduction in relative tumor volume (RTV%) 14 d post-administration and markedly decreased 18F-FDG uptake ( F values: 25.28 and 67.42, t values: 4.64-8.61, P values: 0.001-0.009). Conclusion:The combination of 177Lu-PSMA-I&T and fluzoparib can inhibit prostate cancer cell proliferation and migration, suppress tumor growth and metabolism, and demonstrates synergistic effects more effectively.

Sennoside A(SA),a typical prodrug,exerts its laxative effect only after its transformation into rhei-nanthrone catalyzed by gut microbial hydrolases and reductases.Hydrolases have been identified,but reductases remain unknown.By linking a photoreactive group to the SA scaffold,we synthesized a photoaffinity probe to covalently label SA reductases and identified SA reductases using activity-based protein profiling(ABPP).From lysates of an active strain,Bifidobacterium pseudocatenulatum(B.pseu-docatenulatum),397 proteins were enriched and subsequently identified using mass spectrometry(MS).Among these proteins,chromate reductase/nicotinamide adenine dinucleotide(NADH)phosphate(NADPH)-dependent flavin mononucleotide(FMN)reductase/oxygen-insensitive NADPH nitroreductase(nfrA)was identified as a potent SA reductase through further bioinformatic analysis and The Universal Protein Resource(UniProt)database screening.We also determined that recombinant nfrA could reduce SA.Our study contributes to further illuminating mechanisms of SA transformation to rheinanthrone and simultaneously offers an effective method to identify gut bacterial reductases.

Objective To explore the prognostic value and immune effects of insulin-like growth factor family 2(IGFL2)in bladder cancer based on data from TCGA,TIMER2.0 and HPA.Methods Bioinformatics analysis was conducted to analyze the expression of IGFL2 in bladder cancer and its relationship with survival and prognosis of bladder cancer patients.The receiver operating characteristic(ROC)curve and prognostic nomogram were plotted to analyze the diagnostic value of IGFL2 for bladder cancer and its ability to predict the 5-year overall survival rate of patients.IGFL2-related signaling pathways were analyzed with GSEA enrichment.The correlation between IGFL2 expression and immune cells was analyzed based on data from the TIMER database.Results IGFL2 was highly expressed in bladder cancer(P＜0.001).The high expression of IGFL2 was positively correlated with patients'age,pathological stage,histological grade and T stage(all P＜0.05).Patients'with high IGFL2 expression had shorter overall survival and disease-specific survival than those with low IGFL2 expression(all P＜0.05).IGFL2 could be used as an independent prognostic factor for bladder cancer with good diagnostic and predictive value.GSEA enrichment analysis showed that IGFL2 was involved in signaling pathways related to immunity,inflammation and tumor.High IGFL2 expression was positively correlated with the infiltration of most immune cells.Conclusion IGFL2 is highly expressed in bladder cancer and can be a biomarker for the diagnosis and prognosis of this disease.It may become a potential target for immunotherapy.

AIM:Bone morphogenetic protein 7(BMP7)reduces the expression of Yes-related protein 1(YAP1)by down-regulating Ajuba level and decreasing extracellular matrix(ECM)deposition.This study aimed to inves-tigate the influence of these factors on modifying the degree of renal fibrosis in rats with diabetic nephropathy.METH-ODS:Eighteen Sprague-Dawley(SD)rats were randomly divided into three groups:the normal control(NC)group,the diabetes mellitus(DM)group,and the DM group treated with BMP7 overexpressing adeno-associated virus(DM+rAAV-BMP7).Each group consisted of six rats.Diabetic kidney disease(DKD)was established in the DM and DM+rAAV-BMP7 groups by injecting 55 mg/kg streptozotocin(STZ)via the tail vein.NRK-52E cells were divided into three groups:the normal glucose(NG)group,the high glucose(HG)group,and the high glucose group treated with recombinant hu-man BMP7(HG+rhBMP7)group.Pathological changes in renal tissues were observed using hematoxylin and eosin(HE)and Sirius red staining.Immunohistochemical staining was performed to examine the expression sites of Ajuba and YAP1 in the renal cortex.Western blot analysis was conducted to determine the expression levels of BMP7,Ajuba,YAP1,colla-gen type Ⅲ(Col-Ⅲ),and fibronectin(FN)in the rat renal cortex and NRK-52E cells.RT-qPCR was used to measure the mRNA levels of Ajuba and YAP1 in the rat renal cortex.RESULTS:Biochemical indices revealed significantly ele-vated levels of blood glucose,serum creatinine,triglycerides,total cholesterol,and 24-hour urinary protein in the DM group compared to the NC group(P<0.05).In the DM+rAAV-BMP7 group,the levels of serum creatinine,24-hour uri-nary protein,triglycerides,and total cholesterol were lower than those in the DM group(P<0.05).Pathological staining demonstrated that the renal interstitium of the DM group exhibited inflammatory cell infiltration,fibrous tissue,collagen fi-ber deposition,disordered renal tubule arrangement,atrophy,and vacuolar degeneration,which were ameliorated in the DM+rAAV-BMP7 group.Immunohistochemistry revealed that Ajuba and YAP1 were mainly expressed in the cytoplasm and nucleus,with high expression in the cytoplasm of the DM group,which was significantly decreased in the DM+rAAV-BMP7 group.Western blot results indicated that the protein levels of FN,Col-Ⅲ,Ajuba,and YAP1 were up-regulated in the DM and the HG groups(P<0.05),but significantly down-regulated in the DM+rAAV-BMP7 group(P<0.05).RT-qP-CR results demonstrated that the mRNA levels of Ajuba and YAP1 were higher in the DM group and significantly lower in the DM+rAAV-BMP7 group(P<0.05).CONCLUSION:The overexpression of BMP7 can ameliorate renal fibrosis in rats with DKD.This effect is likely mediated by the down-regulation of Ajuba,reduction of YAP1 expression,and subse-quent inhibition of ECM deposition.

The comprehensive detection and identification of active ingredients in complex matrices is a crucial challenge.Liquid chromatography coupled with high-resolution mass spectrometry(LC-HRMS)is the most prominent analytical platform for the exploration of novel active compounds from complex matrices.However,the LC-HRMS-based analysis workflow suffers from several bottleneck issues,such as trace content of target compounds,limited acquisition for fragment information,and uncertainty in interpreting relevant MS2 spectra.Lycibarbarspermidines are vital antioxidant active ingredients in Lycii Fructus,while the reported structures are merely focused on dicaffeoylspermidines due to their low content.To comprehensively detect the new structures of lycibarbarspermidine derivatives,a"depict"strategy was developed in this study.First,potential new lycibarbarspermidine derivatives were designed according to the biosynthetic pathway,and a comprehensive database was established,which enlarged the coverage of lycibarbarspermidine derivatives.Second,the polarity-oriented sample prep-aration of potential new compounds increased the concentration of the target compounds.Third,the construction of the molecular network based on the fragmentation pathway of lycibarbarspermidine derivatives broadened the comprehensiveness of identification.Finally,the weak response signals were captured by data-dependent scanning(DDA)followed by parallel reaction monitoring(PRM),and the efficiency of acquiring MS2 fragment ions of target compounds was significantly improved.Based on the integrated strategy above,210 lycibarbarspermidine derivatives were detected and identified from Lycii Fructus,and in particular,170 potential new compounds were structurally characterized.The integrated strategy improved the sensitivity of detection and the coverage of low-response components,and it is expected to be a promising pipeline for discovering new compounds.