BACKGROUND:Traditional methods of urinary tract reconstruction are limited by donor scarcity,high complication rates,and suboptimal functional recovery.Tissue engineering strategies offer new directions in this field.Since the urinary tract is mainly composed of muscle tissue,the key is to find suitable seed cells and efficiently induce them to differentiate into smooth muscle cells.Comparative studies on the efficacy of different smooth muscle cell induction regimens are still lacking. OBJECTIVE:To isolate,culture,and identify human urine-derived stem cells,and to compare the effects of two different induction protocols. METHODS:Human urine-derived stem cells were isolated from urine samples of 11 healthy adult volunteers by multiple centrifugations.Surface markers were identified by flow cytometry.The multi-directional differentiation potential of human urine-derived stem cells was verified through osteogenic and adipogenic differentiation.Differentiation was induced by transforming growth factor-β1 or transforming growth factor-β1 combined with platelet derived growth factor for 14 days.Immunofluorescence staining and western blot assay were employed to compare the expression differences of smooth muscle-specific proteins(α-SMA and SM22). RESULTS AND CONCLUSION:(1)Urine-derived stem cells were successfully isolated from the eight urine samples of healthy people.These cells exhibit a"rice grain"-like morphology and possess a robust proliferative capacity.(2)Urine-derived stem cells exhibited high expression of mesenchymal stem cell surface markers(CD73,CD90,and CD44)and extremely low expression of hematopoietic stem cell surface markers(CD34 and CD45).These cells did not express CD19,CD105,and HLA-DR.(3)After osteogenic and adipogenic differentiation,the formation of calcium nodules and lipid droplets was observed,with positive staining results from Alizarin Red S and Oil Red O staining.(4)After 14 days of smooth muscle induction culture,immunofluorescence staining revealed that the smooth muscle differentiation rate of urine-derived stem cells treated with a combination of transforming growth factor-β1 and platelet derived growth factor was significantly higher compared to those treated with transforming growth factor-β1 alone(P<0.005).(5)After 14 days of smooth muscle induction culture,western blot assay further demonstrated that the expression levels of α-SMA and SM22 in the transforming growth factor-β1/platelet derived growth factor group were significantly elevated compared to those in the transforming growth factor-β1 only group(P<0.005).These findings confirm that urine-derived stem cells can be non-invasively isolated using multiple rounds of centrifugation.Compared with transforming growth factor-β1 alone,the combination of transforming growth factor-β1 and platelet derived growth factor can improve the efficiency of inducing urine-derived stem cells to differentiate into smooth muscle cells.

Objective To study the transcriptional expression of Chlorophyti Laxi R.Br.using high-throughput sequencing technology.Methods Total RNA of Chlorophtum laxum R.Br.was extracted,followed by library construction,sequencing,and de novo assembly to obtain unigene sequences.These sequences were then annotated and compared to acquire genetic information of the Chlorophtum laxum R.Br.transcriptome.Results A total of 29 325 unigene were identified,with an average length of 1 026 bp and an N50 of 1 697 bp.Among them,20 338 unigene were functionally annotated in at least one database,with Chlorophtum laxum R.Br.showing the highest sequence similarity to Asparagus officinalis.In the Kyoto Encyclopedia of Genes and Genomes(KEGG)database,13 392 unigene were annotated,including genes involved in flavonoid biosynthesis and ubiquinone and other terpenoid-quinone biosynthesis pathways.Thirty-five upstream genes related to Chlorophtum laxum R.Br.saponin biosynthesis were identified.In the Gene Ontology(GO)database,16 728 unigene were annotated,covering cellular anatomical entities,binding,and biological processes.Using the Microsatellite Identification Tool(MISA),11 486 assembled unigene longer than 1 000 bp were analyzed for simple sequence repeats(SSRs),resulting in the identification of 5 178 SSR loci,for which primers were designed using Primer 3.0.By comparing Chlorophtum laxum R.Br.unigene with the transcription factor database,657 transcription factors,including bHLH,MYB,and WRKY,were identified.Conclusion The transcriptome data provide a foundation for studying the biosynthetic pathways and functional genes of medicinal active components in Chlorophtum laxum R.Br.,and also contribute to the conservation and development of its resources.

This study aims to reveal the effect and mechanism of Gentianella turkestanorum total extract(GTI) in ameliorating non-alcoholic steatohepatitis(NASH). UPLC-Q-TOF-MS was employed to identify the chemical components in GTI. SwissTarget-Prediction, GeneCards, OMIM, and TTD were utilized to screen the targets of GTI components and NASH. The common targets shared by GTI components and NASH were filtered through the STRING database and Cytoscape 3.9.0 to identify core targets, followed by GO and KEGG enrichment analysis. AutoDock was used for molecular docking of key components with core targets. A mouse model of NASH was established with a methionine-choline-deficient high-fat diet. A 4-week drug intervention was conducted, during which mouse weight was monitored, and the liver-to-brain ratio was measured at the end. Hematoxylin-eosin staining, Sirius red staining, and oil red O staining were employed to observe the pathological changes in the liver tissue. The levels of various biomarkers, including aspartate aminotransferase(AST), alanine aminotransferase(ALT), hydroxyproline(HYP), total cholesterol(TC), triglycerides(TG), low-density lipoprotein cholesterol(LDL-C), high-density lipoprotein cholesterol(HDL-C), malondialdehyde(MDA), superoxide dismutase(SOD), and glutathione(GSH), in the serum and liver tissue were determined. RT-qPCR was conducted to measure the mRNA levels of interleukin 1β(IL-1β), interleukin 6(IL-6), tumor necrosis factor α(TNF-α), collagen type I α1 chain(COL1A1), and α-smooth muscle actin(α-SMA). Western blotting was conducted to determine the protein levels of IL-1β, IL-6, TNF-α, and potential drug targets identified through network pharmacology. UPLC-Q-TOF/MS identified 581 chemical components of GTI, and 534 targets of GTI and 1 157 targets of NASH were screened out. The topological analysis of the common targets shared by GTI and NASH identified core targets such as IL-1β, IL-6, protein kinase B(AKT), TNF, and peroxisome proliferator activated receptor gamma(PPARG). GO and KEGG analyses indicated that the ameliorating effect of GTI on NASH was related to inflammatory responses and the phosphoinositide 3-kinase(PI3K)/AKT pathway. The staining results demonstrated that GTI ameliorated hepatocyte vacuolation, swelling, ballooning, and lipid accumulation in NASH mice. Compared with the model group, high doses of GTI reduced the AST, ALT, HYP, TC, and TG levels(P<0.01) while increasing the HDL-C, SOD, and GSH levels(P<0.01). RT-qPCR results showed that GTI down-regulated the mRNA levels of IL-1β, IL-6, TNF-α, COL1A1, and α-SMA(P<0.01). Western blot results indicated that GTI down-regulated the protein levels of IL-1β, IL-6, TNF-α, phosphorylated PI3K(p-PI3K), phosphorylated AKT(p-AKT), phosphorylated inhibitor of nuclear factor kappa B alpha(p-IκBα), and nuclear factor kappa B(NF-κB)(P<0.01). In summary, GTI ameliorates inflammation, dyslipidemia, and oxidative stress associated with NASH by regulating the PI3K/AKT/NF-κB signaling pathway.
Animals ; Non-alcoholic Fatty Liver Disease/genetics* ; Mice ; Network Pharmacology ; Male ; Drugs, Chinese Herbal/administration & dosage* ; Chromatography, High Pressure Liquid ; Liver/metabolism* ; Mice, Inbred C57BL ; Humans ; Mass Spectrometry ; Tumor Necrosis Factor-alpha/metabolism* ; Disease Models, Animal ; Molecular Docking Simulation

ObjectiveTransfer RNA-derived fragments (tRFs) are a recently characterized and rapidly expanding class of small non-coding RNAs, typically ranging from 13 to 50 nucleotides in length. They are derived from mature or precursor tRNA molecules through specific cleavage events and have been implicated in a wide range of cellular processes. Increasing evidence indicates that tRFs play important regulatory roles in gene expression, primarily by interacting with target messenger RNAs (mRNAs) to induce transcript degradation, in a manner partially analogous to microRNAs (miRNAs). However, despite their emerging biological relevance and potential roles in disease mechanisms, there remains a significant lack of computational tools capable of systematically predicting the interaction landscape between tRFs and their target mRNAs. Existing databases often rely on limited interaction features and lack the flexibility to accommodate novel or user-defined tRF sequences. The primary goal of this study was to develop a machine learning based prediction algorithm that enables high-throughput, accurate identification of tRF:mRNA binding events, thereby facilitating the functional analysis of tRF regulatory networks. MethodsWe began by assembling a manually curated dataset of 38 687 experimentally verified tRF:mRNA interaction pairs and extracting seven biologically informed features for each pair: (1) AU content of the binding site, (2) site pairing status, (3) binding region location, (4) number of binding sites per mRNA, (5) length of the longest consecutive complementary stretch, (6) total binding region length, and (7) seed sequence complementarity. Using this dataset and feature set, we trained 4 distinct machine learning classifiers—logistic regression, random forest, decision tree, and a multilayer perceptron (MLP)—to compare their ability to discriminate true interactions from non-interactions. Each model’s performance was evaluated using overall accuracy, receiver operating characteristic (ROC) curves, and the corresponding area under the ROC curve (AUC). The MLP consistently achieved the highest AUC among the four, and was therefore selected as the backbone of our prediction framework, which we named tRF Prospect. For biological validation, we retrieved 3 high-throughput RNA-seq datasets from the gene expression omnibus (GEO) in which individual tRFs were overexpressed: AS-tDR-007333 (GSE184690), tRF-3004b (GSE197091), and tRF-20-S998LO9D (GSE208381). Differential expression analysis of each dataset identified genes downregulated upon tRF overexpression, which we designated as putative targets. We then compared the predictions generated by tRF Prospect against those from three established tools—tRFTar, tRForest, and tRFTarget—by quantifying the number of predicted targets for each tRF and assessing concordance with the experimentally derived gene sets. ResultsThe proposed algorithm achieved high predictive accuracy, with an AUC of 0.934. Functional validation was conducted using transcriptome-wide RNA-seq datasets from cells overexpressing specific tRFs, confirming the model’s ability to accurately predict biologically relevant downregulation of mRNA targets. When benchmarked against established tools such as tRFTar, tRForest, and tRFTarget, tRF Prospect consistently demonstrated superior performance, both in terms of predictive precision and sensitivity, as well as in identifying a higher number of true-positive interactions. Moreover, unlike static databases that are limited to precomputed results, tRF Prospect supports real-time prediction for any user-defined tRF sequence, enhancing its applicability in exploratory and hypothesis-driven research. ConclusionThis study introduces tRF Prospect as a powerful and flexible computational tool for investigating tRF:mRNA interactions. By leveraging the predictive strength of deep learning and incorporating a broad spectrum of interaction-relevant features, it addresses key limitations of existing platforms. Specifically, tRF Prospect: (1) expands the range of detectable tRF and target types; (2) improves prediction accuracy through multilayer perceptron model; and (3) allows for dynamic, user-driven analysis beyond database constraints. Although the current version emphasizes miRNA-like repression mechanisms and faces challenges in accurately capturing 5'UTR-associated binding events, it nonetheless provides a critical foundation for future studies aiming to unravel the complex roles of tRFs in gene regulation, cellular function, and disease pathogenesis.

Objective To explore the clinical efficacy of dual plasma molecular adsorption(DPMAS)sequential plasma exchange(PE)artificial liver mode in the treatment of liver failure(LF).Methods Eighty-five patients with LF receiving the artificial liver treatment in the Affiliated Hospital of Zunyi Medical Univer-sity from January 2020 to December 2023 were selected as the study subjects and divided into the study group(n=52)and the control group(n=33)according to the different treatment modes.The study group conduc-ted DPMAS sequential PE treatment and the control group underwent the PE treatment.The liver function[total bilirubin(TBIL),alanine aminotransferase(ALT),aspartate aminotransferase(AST),serum albumin(ALB),globulin(GLO),prealbumin(PAB)],Hb,coagulation function[platelet(PLT),plasminogen activity(PTA),international normalized ratio(INR),fibrinogen(FIB)]before treatment and at 24 h after treatment were compared between the two groups.Results Compared with before treatment,the levels of TBIL,ALT,AST,GLO and Hb after the first and second treatment in the two groups were decreased,ALB level in the control group and PAB level after the second time treatment was increased(P＜0.05).Compared with after the first treatment,the levels of TBIL,ALT and GLO after the second treatment in the two groups and the levels of AST and Hb in the study group were decreased,ALB level in the study group and PAB level in the two groups were increased(P＜0.05).Compared with before treatment,the levels of PLT and FIB after the first treatment in the two groups and INR level in the control group were decreased,PTA level in the control group was increased(P＜0.05).Compared with before treatment,the levels of PLT,INR and FIB after the second treatment in the two groups were decreased,PTA level was increased(P＜0.05).Compared with be-fore treatment,the levels of PLT,INR and FIB after the second treatment in the two groups were decreased,and PTA level was increased(P＜0.05).Compared with after the first treatment,PTA level after the second treatment in the study group was increased and INR level was decreased.Conclusion PE and DPMAS sequen-tial PE all could improve the liver function in the patients with LF,moreover the two times treatment has more significant effect.

Objective To investigate the effect of field angle on the results of in vivo dose validation of electronic portal imaging device(EPID)in patients.Methods Design the mold test and analyze the influence of different mold thicknesses and different frame angles on the 2D γ pass rate.Twenty-three patients who un-derwent radiotherapy in the Department of Radiotherapy of Jinhua Municipal Central Hospital from January to June 2023 were selected as the research object.In vivo dose verification was carried out during treatment to obtain 2D γ pass rate using same-day sector beam CT(FBCT)of planned CT and executive image guided(IG-RT)as reference images,and the influence of field angle on pass rate was analyzed.Results When the frame angle was unchanged,the area of the shooting field was larger than 17 cm×17 cm,and the 2D γ passing rate decreased with the increase of the thickness.The frame angle had no effect on the 2D γ pass rate when the mold thickness was constant.In clinical treatment data,the passage rate of 2D γ near 0°/180° was higher than that near 90°/270°(P＜0.05),and the passage rate near 90°/270° in the FBCT group was higher than that in the IGRT group(P＜0.05).The median passing rate of 3 mm 2D γ was 97.97％in 3％of the 23 patients.The non-IGRT group was 96.81％,the IGRT group was 97.89％,the FBCT group was 98.94％.There was a statistically significant difference in 2D γ passing rate between the non-IGRT group and the IGRT group(Z=-5.083,P＜0.05),and there was a statistically significant difference in 2D γ passing rate between the IGRT group and the FBCT group(Z=-10.657,P＜0.05).Conclusion Clinically,the difference of pass rate in vi-vo dose verification at different rack angles is mainly due to the difference of images within and between ses-sions.Using same-day FBCT as the reference image for in-vivo dose verification can improve the accuracy of pass rate and eliminate the influence of image difference between sessions.

Objective:To observe the impact of ultra early neurorehabilitation combined with cognitive intervention training on the psychological state,neurological function,and quality of life of stroke patients.Methods:89 stroke patients who were admitted to our hospital from April 2023 to May 2024 were divided into control group(conventional rehabilitation intervention combined with cognitive intervention training,44 cases)and study group(control group combined with ultra early neurological rehabilitation,45 cases)by using random number table method.The neurological function,daily self-care ability,psychological state,quality of life,and satisfaction between two groups were compared.Results:14 d after intervention,the study group had lower scores on the national institutes of health stroke scale(NIHSS),self rating anxiety scale(SAS),and self rating depression Scale(SDS)compared to the control group,had higher scores of the comprehensive quality of life evaluation questionnaire-74(CQOLI-74)and the modified barthel index(MBI)(P＜0.05).The overall satisfaction rate of the study group was higher than that of the control group(P＜0.05).Conclusion:Ultra early neurorehabilitation combined with cognitive intervention training can effectively improve the neurological function and daily self-care ability of stroke patients,reduce anxiety and depression,improve their quality of life,and achieve good patient satisfaction.

Objective:To understand the current status of knowledge, attitude, and practice (KAP) of nurses in palliative care under the model of palliative co-care in hospitals of different levels in Zhejiang Province, and to analyze the influencing factors.Methods:This study was a cross-sectional study. Convenience sampling was used to select 541 clinical registered nurses from 3 Class Ⅲ hospitals, 3 Class Ⅱ hospitals, and 2 ClassⅠ hospitals in Hangzhou, Huzhou, and Lishui, Zhejiang Province, from March to April 2023. A general information questionnaire and the KAP Survey Scale of Palliative Care Under the Palliative Co-care Model were used to investigate the subjects. Multiple linear regression analysis was conducted to analyze the influencing factors of nurses' KAP of palliative care under palliative co-care. A total of 541 questionnaires were distributed, and 535 valid questionnaires were recovered, with an effective response rate of 98.89%.Results:The scores of 535 nurses for palliative care knowledge, attitude, and practice were (16.29±1.99), (71.10±9.00), and (32.21±5.89), respectively. Multiple linear regression analysis revealed that hospital level, gender, and whether nurses have participated in on-the-job training on palliative care are influencing factors of nurses' knowledge of palliative care ( P<0.05) ; ethnicity and the duration of palliative care training courses attended within two years are influencing factors of nurses' attitude toward palliative care ( P<0.05) ; department is an influencing factor of nurses' practice of palliative care ( P<0.05) . Conclusions:Under the palliative co-care model in Zhejiang Province, nurses demonstrate a good attitude toward palliative care, while their knowledge and practice levels of palliative care still have room for improvement, with distinct influencing factors for each dimension-hospital level, gender, and on-the-job training participation affect knowledge; ethnicity and training duration within two years affect attitude; and department affects practice.

BACKGROUND:Previous studies have shown that Huosui Formula has a synergistic effect on the immune and hematopoietic regulation of patients with myelodysplastic syndrome,but the specific mechanism is not yet clear.OBJECTIVE:To explore the effect and mechanism of Huosui Formula on bone marrow hematopoiesis in rats with myelodysplastic syndrome.METHODS:A total of 70 SD rats were randomly divided into a normal control group(n=10),a model group(n=15),a western medicine group(n=15),a low-dose Huosui Formula group(n=15),and a high-dose Huosui Formula group(n=15).Except for the normal control group,the other four groups were injected with dimethyl benzanthracene via the tail vein to induce the establishment of rat myelodysplastic syndrome models.After modeling,the normal control group and the model group were given normal saline;the western medicine group was given thalidomide capsules 10 mg/kg and retinoic acid tablets 4 mg/kg,and the low-dose Huosui Formula group and the high-dose Huosui Formula group were given 1.5 and 6 g/kg Huosui Formula,respectively,by intragastric administration once a day for 28 consecutive days.Peripheral blood and femoral bone marrow tissue were collected to detect peripheral blood routine and bone marrow biopsy hematopoietic proliferation.Flow cytometry was used to detect T lymphocyte subsets and the expression of CTLA-4 and PD-1 on T lymphocytes.RESULTS AND CONCLUSION:(1)Compared with the normal control group,peripheral blood leukocyte,neutrophil,hemoglobin,platelet,and CD4+,CD4+/CD8+levels were decreased in the model group significantly(P＜0.05),while CD4+PD-1+,CD8+PD-1+,CD4+CTLA-4+,and CD8+CTLA-4+expressions were significantly upregulated(P＜0.05).(2)In all dosage groups,myelopoietic proliferation was increased compared with the model group,with no significant difference between the groups(P＞0.05).(3)Compared with the model group,leukocytes,hemoglobin,platelets,and CD4+,CD4+/CD8+were significantly elevated in the high-dose Huosui Formula group(P＜0.05),the expression of CD8+was significantly lower(P＜0.05),and the levels of CD4+PD-1+,CD8+PD-1+,CD4+CTLA-4+,and CD8+CTLA-4+were down-regulated but not statistically significant(P＞0.05).(4)The western medicine group and the high-dose Huosui Formula group showed similar efficacy.The improvement of each index in the high-dose Huosui Formula group was superior to that in the low-dose Huosui Formula group.These findings indicate that Huosui Formula can improve the bone marrow hematopoiesis in myelodysplastic syndrome model rats,increase the levels of CD4+,and CD4+/CD8+while down-regulate the expression levels of CD4+PD-1+,CD8+PD-1+,CD4+CTLA-4+,and CD8+CTLA-4+.These observations suggest a link to the negative immunoregulation mechanism.