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.

ObjectiveTo investigate the therapeutic effect of Astragali Radix-mediated changes in gut microbiota on treating type 2 diabetes （T2DM）. MethodsA 12-week randomized， placebo-controlled clinical trial enrolled eighty patients with newly diagnosed type 2 diabetes and poor glycemic control in the Qi deficiency type. All patients received insulin therapy. The observation group （40 cases） was administered with Astragali Radix Granules， while the control group （40 cases） received a placebo. Both treamtents were taken orally twice daily. Changes in gut microbiota were assessed by 16s rDNA sequencing. Serum glucagon-like peptide-1 （GLP-1） levels were measured using enzyme-linked immunosorbent assay （ELISA）. Glucose metabolism indicators including fasting blood glucose （FPG）， 2-hour postprandial blood glucose （2 h PG），glycated albumin（GA）， and glycated hemoglobin （HbA1c） were evaluated. Pancreatic function was evaluated using fasting C-peptide （FCP）， 2-hour postprandial C-peptide （2 h CP）， and C-peptide area under the curve （AUC_cp）. Traditional Chinese medicine （TCM） syndrome scores， clinical efficacy， and safety indicators were also observed. ResultsIn terms of glucose metabolism indicators， compared with the baseline， both groups exhibited significantly lower FPG， 2 h PG， GA and HbA1C （P<0.01），while FCP， 2 h CP and AUC_cp were significantly higher （P<0.01）. Compared with the control group after the treatment， the observation group showed significantly lower FPG， 2 h PG， GA and HbA1C（P<0.05， P<0.01），and significantly higher FCP， 2 h CP and AUC_cp （P<0.05， P<0.01）， indicating that Astragali Radix can improve glucose metabolism. In terms of the diversity of gut microbiota， no significant differences were detected in the Chao1， Shannon and Simpson indexes of the two groups compared with their respective baselines. However， compared with the post-treatment control group， the observation group demonstrated significant increases in the Chao1， Shannon and Simpson indexes （P<0.05， P<0.01）. The β-diversity analysis showed significant separation in gut microbiota composition before and after treatment in both groups， indicating that Astragali Radix can significantly alter the structure and improve the diversity of gut microbiota. At the phylum level， compared with the baseline， both groups showed a significant increase in the relative abundance of Bacteroidota（P<0.01）. The relative abundance of the potentially harmful phylum Proteobacteria was significantly lower in the observation Group after treatment （P<0.01）. Compared with the post-treatment control group， the observation group had a significantly higher relative abundance of Bacteroidota（P<0.01）. No significant difference was found in Firmicutes/Bacteroidota （F/B） ratio between the two groups after treatment， and other phyla showed no significant differences. At the genus level， compared with the baseline， the observation group exhibited a significant increase in Bacteroides （P<0.01） and a significant decrease in Escherichia-Shigella （P<0.01）， whereas no significant difference was seen in the control group . Compared with the control group after treatment， the observation group after treatment had a significantly higher relative abundance of Bacteroides （P<0.01）. No significant differences were seen in other genera. Linear discriminant analysis （LDA） identified potential characteristics taxa： in the observation group， Bacteroidota at the phylum level and Bacteroides and Dubosiella at the genus level， in the control group， Proteobacteria at the phylum level as well as Barnesiella and Staphylococcus at the genus level. Correlation analysis based on a heatmap revealed that GLP-1 levels were positively correlated with Firmicutes， F/B ratio and Fusobacterium， and negatively correlated with Bacteroidota， Proteobacteria， Bacteroides and Escherichia-Shigella. In terms of clinical efficacy， compared with the control group， the total effective rate of the observation group was significantly higher （P<0.05）. Compared with the baseline， the scores for shortness of breath， fatigue， weakness， spontaneous sweating and reluctance to speak significantly decreased in both groups （P<0.01）. Compared with the control group after treatment， the score for weakness was significantly lower in the observation group （P<0.01），indicating that Astragali Radix could improve clinical symptoms and alleviate weakness symptoms. In terms of safety， compared with the baseline， alanine aminotransferase （ALT） levels significantly decreased in both groups （P<0.05，P<0.01），indicating that Astragali Radix did not induce any significant abnormalities in liver and kidney functions. ConclusionAstragali Radix demonstrates the potential to significantly improve the gut microbiota environment in patients of newly diagnosed type 2 diabetes with Qi deficiency. The therapeutic effect may contribute to glycemic control， possibly mediated by an elevation in GLP-1 level. These findings may support its further clinical investigations and potential applications.

Objective To explore the pathogenesis and prognostic factors of liposarcoma in the head and neck region, and simultaneously analyze the efficacy of different treatment regimens. Methods A retrospective analysis was performed on all patients with primary untreated head and neck liposarcoma who were diagnosed and underwent surgical treatment at our hospital from January 2008 to January 2024. All patients were monitored during follow-up, and their prognoses were analyzed using SPSS software. Results A total of 30 patients were included in the study. Liposarcoma accounted for up to 60% of the cases in the orbit, while the remaining liposarcomas were primarily located in various interspaces of the neck. Dedifferentiated liposarcoma was the most common type, comprising 33%, while myxoid pleomorphic liposarcoma was the rarest at 4%. The tumor pathological type (P<0.001) and Ki67 (P=0.014) significantly affected the tumor control rate. However, an analysis of disease-specific survival rates revealed no significant differences across various factors (all P>0.05). Conclusion The prognosis of head and neck liposarcoma is better compared to that of liposarcomas in other parts of the body. However, myxoid pleomorphic liposarcoma, pleomorphic fat sarcoma, and high Ki67 levels are indicators of poor prognosis. Additionally, postoperative adjuvant radiotherapy does not significantly enhance disease-specific survival rates.

Objective:To explore the potential risk factors for cancer in patients with adenomatous polyps based on tumor markers and polyp lesion characteristics.Methods:A retrospective analysis was conducted to collect clinical data of 115 patients with adenomatous intestinal polyps who visited Nanjing Jiangbei Hospital from November 2022 to November 2024. They were divided into a cancerous group (17 cases) and a non cancerous group (98 cases) based on whether they were cancerous or not. Clinical data such as tissue type and polyp site and tumor marker levels such as carcinoembryonic antigen (CEA) and cancer antigen 72-4 (CA72-4) were collected at the first visit of all patients. The potential risk factors of adenomatous intestinal polyp canceration were investigated by Logistic regression analysis.Results:Univariate analysis revealed that the proportion of villous tubular adenomas, central depression of polyps, and lobulated polyps in the cancerous group were higher than those in the non cancerous group. Serum levels of CEA and CA72-4 were also higher in the cancerous group than in the non cancerous group : 13/17 vs.47.96% (47/98), 7/17 vs. 15.31% (15/98), 6/17 vs. 8.16% (8/98), (6.41 ± 1.81) μg/L vs. (4.23 ± 1.48) μg/L, (6.98 ± 1.83) kU/L vs. (5.66 ± 1.78) kU/L, respectively. The difference was statistically significant ( P<0.05). The results of Logistic regression analysis showed that the histological subtype of villous tubular adenoma, central depression of polyps, lobulated polyps, and high levels of CEA and CA72-4 were independent risk factors for cancer in patients with adenomatous intestinal polyps ( P<0.05). A nomogram risk model was constructed based on the influencing factors of canceration in patients with adenomatous intestinal polyps. The calibration curve was drawn, and the calibration curve was similar to the Y-X straight line, suggesting that the evaluation results of the nomogram risk model were highly consistent with the actual observation results. The receiver operating characteristic (ROC) curve was drawn. The results showed that the area under the curve (AUC) of the nomogram risk model for evaluating the canceration of patients with adenomatous intestinal polyps was 0.956, and the evaluation value was high. The decision curve was drawn, with the threshold of high risk as the horizontal coordinate and the net rate of return as the vertical coordinate. The results showed that when the threshold was in the range of 0 - 0.85, 0.96 - 0.99, the net benefit rate of predicting the cancer risk of patients with adenomatous intestinal polyps was greater than 0 and the maximum net benefit rate was 0.148. Conclusions:The histological classification of villous tubular adenoma, central depression of polyps, lobulated polyps, and high levels of CEA and CA72-4 are independent risk factors for cancer in patients with adenomatous intestinal polyps; The evaluation efficiency of the column chart risk model constructed based on the above factors is good.

Objective To explore the status and trajectory of urinary incontinence in patients with bladder cancer after orthotopic neobladder surgery,and analyze the characteristics of different trajectory categories,to provide references for the intervention and management of urinary incontinence in patients after orthotopic neobladder surgery.Methods A prospective study design was adopted,and the convenience sampling was used to select 142 eligible patients who underwent in orthotopic neobladder surgery in a tertiary hospital from January 2019 to August 2022.Demographic and clinical data of the patients were collected.The continence status of patients was evaluated by the International Consultation on Incontinence Questionnaire-Short Form(ICI-Q-SF)at 1 month after surgery,3 months after surgery,6 months after surgery and 1 year after surgery.Additionally,postoperative physical condition scores,pelvic floor muscle training,and timed urination information were collected.The Group-Based Trajectory Model was used to identify the development trajectory of urinary incontinence,and the logistic regression was conducted to identify influencing factors of urinary incontinence trajectories.Results 142 cases were included.There are 3 distinct daytime and nocturnal incontinence trajectory groups that were respectively identified:the significantly improved group(21.83％,29.58％),the slowly alleviated group(52.82％,48.59％),and the risk fluctuation group(25.35％,21.83％).Multivariate logistic regression analysis indicated that age,preservation of neurovascular bundles,and adherence to timed voiding were factors significantly influencing urinary incontinence trajectory categories(P＜0.05).Conclusion The trajectory of urinary incontinence in patients undergoing orthotopic neobladder surgery showed an overall improvement trend within a year,with group differences.Clinical attention should be paid to the dynamic evaluation of postoperative urinary incontinence in older,unreserved nerves,and those who did not adhere to scheduled urination,and a phased matching postoperative rehabilitation plan should be developed to improve the urinary control level of patients.

Objective To study the effect and molecular mechanism of vitamin D(VitD)on Tfh cells of MRL/lpr lupus mice.Methods C57/B6 mice and MRL/lpr lupus mice were transfected with siRNA to construct VDR knockout mouse models.Splenic Tfh cells of C57/B6 mice and MRL/lpr lupus mice were divided into control group,lupus group and VDRsiRNA lupus group(treated with vitamin D 0,1 and 10 nmol·L-1,respectively)by siRNA transfection.The percentage of Tfh cells was detected by flow cytometry.MRL/lpr lupus mice Peyer node Tfh cells were randomly divided into 7 groups,blank control group,vitamin D dose groups of 1 and 10 nmol·L-1,paricalcitol group(VitD 10 nmol·L-1+PA),VDRsiRNA control group,VDRsiRNA group(VitD 10 nmol·L-1),CaN inhibitor group(VitD 10 nmol·L-1+CsA),and incubated for 72 h.The concentration of calcium ions in Tfh of each group was detected.The expressions of AT1R,NFAT,CaN and P-CaN in Tfh cells were determined by Western blotting.Results The percentage of Tfh cells decreased significantly with the increase of vitamin D dose.Vitamin D CaN reduce the intracellular calcium concentration of Tfh,up-regulate the expression of AT1 protein in Tfh cells,and down-regulate the expression of CAN,P-CaN and NFAT protein in a dose-dependent manner,and the effect is more obvious when combined with PA.Conclusion Vitamin D may regulate the activation of follicular T helper cells in MRL/lpr mice via the Ca-CaN-NFAT pathway.

[Objective]To explore the general laws of the occurrence and development of the main prescription classification methods.[Methods]Based on the classification concepts of various formulas and the era background of the corresponding periods,explore the research for the emergence and development of disease classification,function classification,species classification and stroke classification.[Results]Disease classification was the main classification approach employed in medical books of past dynasties,which originated from the need of physicians to popularize their medication experience.Function classification and species classification were mainly refined during the Ming and Qing Dynasties.The common material basis for their emergence and development was the substantial number of proven prescriptions accumulated during the course of the development of traditional Chinese medicine.Among them,the function classification was used for organizing and researching current prescriptions,while the species classification was more frequently applied in the study of classical prescriptions from Treatise on Febrile Diseases.[Conclusion]Disease classification constitutes the main thread in the development of formula classification,profoundly mirroring the evolution of the overall theory of traditional Chinese medicine,which serves as the principal vehicle for the inheritance of the medicinal application experience in traditional Chinese medicine and directly caters to clinical practice.Function classification and species classification are further derivations of the classification of diseases and syndromes in the two trajectories of current prescriptions and classical prescriptions,both leaning towards theory to a greater extent.

Objective To explore the status and trajectory of urinary incontinence in patients with bladder cancer after orthotopic neobladder surgery,and analyze the characteristics of different trajectory categories,to provide references for the intervention and management of urinary incontinence in patients after orthotopic neobladder surgery.Methods A prospective study design was adopted,and the convenience sampling was used to select 142 eligible patients who underwent in orthotopic neobladder surgery in a tertiary hospital from January 2019 to August 2022.Demographic and clinical data of the patients were collected.The continence status of patients was evaluated by the International Consultation on Incontinence Questionnaire-Short Form(ICI-Q-SF)at 1 month after surgery,3 months after surgery,6 months after surgery and 1 year after surgery.Additionally,postoperative physical condition scores,pelvic floor muscle training,and timed urination information were collected.The Group-Based Trajectory Model was used to identify the development trajectory of urinary incontinence,and the logistic regression was conducted to identify influencing factors of urinary incontinence trajectories.Results 142 cases were included.There are 3 distinct daytime and nocturnal incontinence trajectory groups that were respectively identified:the significantly improved group(21.83％,29.58％),the slowly alleviated group(52.82％,48.59％),and the risk fluctuation group(25.35％,21.83％).Multivariate logistic regression analysis indicated that age,preservation of neurovascular bundles,and adherence to timed voiding were factors significantly influencing urinary incontinence trajectory categories(P＜0.05).Conclusion The trajectory of urinary incontinence in patients undergoing orthotopic neobladder surgery showed an overall improvement trend within a year,with group differences.Clinical attention should be paid to the dynamic evaluation of postoperative urinary incontinence in older,unreserved nerves,and those who did not adhere to scheduled urination,and a phased matching postoperative rehabilitation plan should be developed to improve the urinary control level of patients.

[Objective]To explore the general laws of the occurrence and development of the main prescription classification methods.[Methods]Based on the classification concepts of various formulas and the era background of the corresponding periods,explore the research for the emergence and development of disease classification,function classification,species classification and stroke classification.[Results]Disease classification was the main classification approach employed in medical books of past dynasties,which originated from the need of physicians to popularize their medication experience.Function classification and species classification were mainly refined during the Ming and Qing Dynasties.The common material basis for their emergence and development was the substantial number of proven prescriptions accumulated during the course of the development of traditional Chinese medicine.Among them,the function classification was used for organizing and researching current prescriptions,while the species classification was more frequently applied in the study of classical prescriptions from Treatise on Febrile Diseases.[Conclusion]Disease classification constitutes the main thread in the development of formula classification,profoundly mirroring the evolution of the overall theory of traditional Chinese medicine,which serves as the principal vehicle for the inheritance of the medicinal application experience in traditional Chinese medicine and directly caters to clinical practice.Function classification and species classification are further derivations of the classification of diseases and syndromes in the two trajectories of current prescriptions and classical prescriptions,both leaning towards theory to a greater extent.

The amino-terminal pro-C-type natriuretic peptide(NT-proCNP)is a diagnostic inflam-matory marker clinically used for diagnosing bacterial infections.This study aims to establish a phage display library of single-chain variable fragment(scFv)antibodies against canine NT-proC-NP and to screen for scFvs with high binding affinity to NT-proCNP.Initially,NT-proCNP was prepared using prokaryotic expression system and was used to immunize New Zealand White rab-bits.Upon achieving the desired serum titer,total RNA was extracted from the splenocytes of rab-bits and reverse transcribed into cDNA.Using this cDNA as a template,degenerate primers were employed to amplify the genes of the rabbit antibody light chain variable region(VL)and heavy chain variable region(VH).The VL and VH regions were spliced together to form a complete scFv fragment via overlap extension PCR.The scFv was then ligated into the phagemid pComb3XSS and electroporated into competent E.coli TG1 cells to construct a rabbit-derived anti-NT-proCNP scFv immunological library.This library underwent four rounds of enrichment and screening to isolate specific single-chain antibodies.The selected antibody was subsequently ex-pressed in a soluble form within a prokaryotic system,and its immunological activity was evalua-ted.Using phage display technology,this study successfully identified a single-chain antibody scFv-1-CNP with strong antigen-binding activity and genetic sequence characteristics of scFvs,providing a research direction for further exploration of scFv applications in the detection of NT-proCNP.