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.

Anthoceros angustus Steph. is rich in phenolic acids such as rosmarinic acid (RA). Phenylalanine ammonia-lyase (PAL) is an entry enzyme in the phenylpropanoid pathway of plants, playing an important role in the biosynthesis of RA. To investigate the important role of PAL in rosmarinic acid synthesis, two PAL genes (designated as AanPAL1 and AanPAL2) were cloned from A. angustus, encoding 755 and 753 amino acid residues, respectively. The AanPAL deduced amino acid sequences contain the conserved domains of PAL and the core active amino acid residues Ala-Ser-Gly. The phylogenetic analysis indicated that AanPAL1 and AanPAL2 were clustered with PALs from bryophytes and ferns and had the shortest evolutionary distance with the PALs from Physcomitrella patens. Quantitative real-time PCR results showed that the expression of AanPAL1 and AanPAL2 was induced by exogenous methyl jasmonate (MeJA). HPLC results showed that the MeJA treatment significantly increased the accumulation of RA. AanPAL1 and AanPAL2 were expressed in Escherichia coli and purified by histidine-tag affinity chromatography. The recombinant proteins catalyzed the conversion of L-phenylalanine to generate trans-cinnamic acid with high efficiency, with the best performance at 50 ℃ and pH 8.0. The K_m and k_cat of AanPAL1 were 0.062 mmol/L and 4.35 s^-1, and those of AanPAL2 were 0.198 mmol/L and 14.48 s^-1, respectively. The specific activities of AanPAL1 and AanPAL2 were 2.61 U/mg and 8.76 U/mg, respectively. The two enzymes had relatively poor thermostability but good pH stability. The high activity of AanPAL2 was further confirmed via whole-cell catalysis with recombinant E. coli, which could convert 1 g/L L-phenylalanine into trans-cinnamic acid with a yield of 100% within 10 h. These results give insights into the regulatory role of AanPAL in the biosynthesis of RA in A. angustus and provide candidate enzymes for the biosynthesis of cinnamic acid.
Phenylalanine Ammonia-Lyase/metabolism* ; Cloning, Molecular ; Cinnamates/metabolism* ; Recombinant Proteins/metabolism* ; Rosmarinic Acid ; Depsides/metabolism* ; Escherichia coli/metabolism* ; Amino Acid Sequence ; Plant Proteins/metabolism* ; Phylogeny ; Acetates/pharmacology* ; Cyclopentanes ; Oxylipins

Objective To explore the effects of different nerve blocks under ultrasound guidance on the expression of inflammatory factors after total hip replacement in the elderly,and its correlation with postoperative cognitive dysfunction(POCD).Methods A total of 100 elderly patients who underwent total hip replacement in our hospital from February to November 2023 were selected as the research objects,and they were divided into the femoral nerve block(FNB)group and the pericapsule nerve group block(PNGB)group according to different nerve block methods during the operation.The expression of inflammatory factors such as serum interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-10(IL-10),tumor necrosis factor-α(TNF-α)and tumor necrosis factor-β(TNF-β)1 day before surgery,6 hours after anesthesia,1 day after surgery,and 3 days after surgery of patients in the two groups were compared.The cognitive function of patients was evaluated by the mini-mental state examination(MMSE)score,and the pain of patients was evaluated by visual analogue scale(VAS)score.GEE model was used to evaluate the effects of two kinds of nerve block on the expression of inflammatory factors.Logistic regression was used to analyzed the correlation between postoperative inflammatory factors and POCD in patients.Results The levels of IL-1β,IL-6,IL-10,TNF-α and TNF-β 1 day after operation in the PNGB group were higher than those in the FNB group,and the differences were statistically significant(P<0.05).The MMSE score 1 day after surgery in the PNGB group was significantly higher than that in the FNB group(P<0.05),and the VAS score 3 days after surgery was significantly lower than that in the FNB group(P<0.05).GEE model showed that PNGB has a greater impact on IL-1β,IL-6,IL-10,TNF-α,and TNF-β.IL-1β,IL-6,IL-10,TNF-α,and TNF-β were all independently related to POCD induced by two different nerve blocks(P<0.05).Conclusion PNGB can effectively inhibit pro-inflammatory factors,reduce inflammatory stress responses,and maintain the balance of inflammatory factors.Inflammatory factors are independently related to POCD induced by different nerve blocks.With the occurrence of inflammatory stress response,the risk of POCD in patients increases.

Objective To analyze the efficacy of vascularized lymph node transplantation(VLNT)combined with lymphatic venous anastomosis(LVA)in the treatment of secondary lower extremity lymphedema assisted by super microsurgical techniques.Methods A retrospective analysis was performed for 15 patients with secondary lower limb lymphedema who underwent VLNT+LVA surgery in the Department of Reconstructive and Reconstructive Microsurgery of Ruikang Hospital Affiliated to Guangxi University of Traditional Chinese Medicine from July 2021 to July 2023,and compared the circumference and volume of each segment of the lower limb between the preoperative and postoperative 90 days.LVA according to the results of ICG examination,3-5 parts of the lower limb were selected for"Z"shaped surgery,and the lymphatic vessels and venules were anastomosed under the microscope in the subcutaneous fat layer.VLNT confirmed and labeled the saphenous branch of the descending knee artery and its accompanying veins in the popliteal fossa,and the latissimus dorsi lymph node flap was incised,and the flap vessels were anastomosed with the saphenous branch of the descending knee artery and its accompanying veins.Postoperative observation of flap vascularization.The circumference and volume of the affected limb were measured before surgery and 90 days after surgery.Results A total of 15 patients with lower extremity lymphedema were included without serious complications.Statistical analysis showed that the circumference of all levels of the affected limb and the volume of the affected limb were improved 90 days after operation compared with those before surgery.Among them,the limb circumference and volume at each level from the highest point of the dorsum of the foot to 52 cm above the ankle improved 90 days after the operation compared with those before the operation.Conclusion LNT+LVA treatment for secondary lower extremity lymphedema can effectively control edema and improve the function of the affected limb.

AIM To investigate the ameliorative effects of Compound Fufangteng Mixture(CFM)on cyclophosphamide(CTX)-induced immunosuppression in mice.METHODS Forty-eight male C57BL/6J mice were randomly divided into the blank control group,the model group,the levamisole hydrochloride group(40 mg/kg)and the low-dose,medium-dose and high-dose CFM groups(3.75,7.5,10 g/kg),with 8 mice in each group,and given respective intervention orally once daily for 14 days.On the 5th to 7th day of administration,with the blank control group given normal saline intraperitoneally,the other groups underwent intraperitoneal CTX injections(80 mg/kg).24 hours after the last administration,organ indices of thymus and spleen were calculated;splenic histopathological alterations were assessed by HE staining;serum levels of IL-2,IL-6 and IgG were quantified using ELISA;splenic CD4+,CD8+T lymphocytes,alongside CD86+and CD206+macrophages populations were analyzed by flow cytometry;and splenic expression of CD4,CD8 and F4/80 was evaluated by immunohistochemical staining.RESULTS In CTX-treated mice,CFM administration mitigated body weight loss;enhanced thymus weight and thymic index;ameliorated splenic immune cell populations,elevated serum levels of cytokines IL-2,IL-6 and IgG in serum;and upregulated splenic levels of CD45+CD3+T lymphocytes and F4/80+CD11b+macrophages,alongside increasing the expression of CD4,CD8 and F4/80 surface markers.CONCLUSION CFM alleviates CTX-induced immunosuppression state in mice by modulating immune cells,restoring immune function and enhancing anti-inflammatory and tissue repair capabilities.

Objective To explore the effects of different nerve blocks under ultrasound guidance on the expression of inflammatory factors after total hip replacement in the elderly,and its correlation with postoperative cognitive dysfunction(POCD).Methods A total of 100 elderly patients who underwent total hip replacement in our hospital from February to November 2023 were selected as the research objects,and they were divided into the femoral nerve block(FNB)group and the pericapsule nerve group block(PNGB)group according to different nerve block methods during the operation.The expression of inflammatory factors such as serum interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-10(IL-10),tumor necrosis factor-α(TNF-α)and tumor necrosis factor-β(TNF-β)1 day before surgery,6 hours after anesthesia,1 day after surgery,and 3 days after surgery of patients in the two groups were compared.The cognitive function of patients was evaluated by the mini-mental state examination(MMSE)score,and the pain of patients was evaluated by visual analogue scale(VAS)score.GEE model was used to evaluate the effects of two kinds of nerve block on the expression of inflammatory factors.Logistic regression was used to analyzed the correlation between postoperative inflammatory factors and POCD in patients.Results The levels of IL-1β,IL-6,IL-10,TNF-α and TNF-β 1 day after operation in the PNGB group were higher than those in the FNB group,and the differences were statistically significant(P<0.05).The MMSE score 1 day after surgery in the PNGB group was significantly higher than that in the FNB group(P<0.05),and the VAS score 3 days after surgery was significantly lower than that in the FNB group(P<0.05).GEE model showed that PNGB has a greater impact on IL-1β,IL-6,IL-10,TNF-α,and TNF-β.IL-1β,IL-6,IL-10,TNF-α,and TNF-β were all independently related to POCD induced by two different nerve blocks(P<0.05).Conclusion PNGB can effectively inhibit pro-inflammatory factors,reduce inflammatory stress responses,and maintain the balance of inflammatory factors.Inflammatory factors are independently related to POCD induced by different nerve blocks.With the occurrence of inflammatory stress response,the risk of POCD in patients increases.

Aim To investigate the effects of m6A demethylase ALKBH5 on cardiomyocytes pyroptosis in mice with myocardial infarction(MI).Methods The MI model of left anterior descending coronary artery ligation surgery was established by knocking down ALKBH5 using adeno-associated virus,and the hypox-ia model of mouse cardiomyocytes(HL-1)was estab-lished by knocking down small interfering RNA.The effects of ALKBH5 on the pyroptosis of MI mice and hypoxic HL-1 cells were observed.Subsequently,mechanism studies were conducted at the cellular lev-el,and the binding of ALKBH5 and IGF2BP2 to NL-RP3 mRNA was detected through RNA pull down and RNA immunoprecipitation(RIP)experiments.The MeRIP-qPCR method was used to determine the effects of ALKBH5 on the mRNA m6A level of NLRP3.Acti-nomycin D for RNA stability experiments were conduc-ted to detect the effects of ALKBH5 and IGF2BP2 on the stability of NLRP3 mRNA.Results Knocking down ALKBH5 in vivo and in vitro both inhibited NL-RP3 inflammasome activation and alleviated pyroptosis in MI mice and hypoxic HL-1 cells.Mechanistically,the results showed that NLRP3 mRNA could bind to ALKBH5 protein in HL-1 cells;knocking down ALK-BH5 could increase the m6A level of NLRP3 and re-duce the stability of NLRP3 mRNA;subsequently,it was confirmed that NLRP3 mRNA and IGF2BP2 pro-tein bound to each other;knocking down IGF2BP2 in-creased the mRNA stability of NLRP3.The Rescue ex-periment showed that knocking down IGF2BP2 re-versed the decrease in NLRP3 mRNA expression caused by knocking down ALKBH5.Conclusions ALKBH5 mediated m6A modification of NLRP3 pro-motes cardiomyocytes pyroptosis in mice with myocardi-al infarction.

AIM To investigate the ameliorative effects of Compound Fufangteng Mixture(CFM)on cyclophosphamide(CTX)-induced immunosuppression in mice.METHODS Forty-eight male C57BL/6J mice were randomly divided into the blank control group,the model group,the levamisole hydrochloride group(40 mg/kg)and the low-dose,medium-dose and high-dose CFM groups(3.75,7.5,10 g/kg),with 8 mice in each group,and given respective intervention orally once daily for 14 days.On the 5th to 7th day of administration,with the blank control group given normal saline intraperitoneally,the other groups underwent intraperitoneal CTX injections(80 mg/kg).24 hours after the last administration,organ indices of thymus and spleen were calculated;splenic histopathological alterations were assessed by HE staining;serum levels of IL-2,IL-6 and IgG were quantified using ELISA;splenic CD4+,CD8+T lymphocytes,alongside CD86+and CD206+macrophages populations were analyzed by flow cytometry;and splenic expression of CD4,CD8 and F4/80 was evaluated by immunohistochemical staining.RESULTS In CTX-treated mice,CFM administration mitigated body weight loss;enhanced thymus weight and thymic index;ameliorated splenic immune cell populations,elevated serum levels of cytokines IL-2,IL-6 and IgG in serum;and upregulated splenic levels of CD45+CD3+T lymphocytes and F4/80+CD11b+macrophages,alongside increasing the expression of CD4,CD8 and F4/80 surface markers.CONCLUSION CFM alleviates CTX-induced immunosuppression state in mice by modulating immune cells,restoring immune function and enhancing anti-inflammatory and tissue repair capabilities.

BACKGROUND:In recent years,postmenopausal osteoporosis has been widely concerned by the society.With the gradual deepening of the research on animal models,animal models have become an important means of modern traditional Chinese medicine research as well as the experimental basis.Therefore,it is particularly important to understand the current status,hotspots,and development trend of the research on animal models.OBJECTIVE:To analyze the research status,hotspots,and development trend of animal models of postmenopausal osteoporosis at home and abroad,and to provide a certain theoretical foundation and reference basis for subsequent research.METHODS:Using"Subject=Postmenopausal osteoporosis AND Subject line=Mouse+Rabbit+Dog+Pig+Sheep+Monkey+Fish+Laboratory animals+Animal experimentation"in Chinese as the search formula,we searched for relevant literature published from January 1,1999 to October 1,2023 in CNKI and WanFang databases.Meanwhile,the articles published from January 1,1999 to October 1,2023 in the Web of Science Core Collection database were searched using"TS=(Postmenopausal osteoporosis)AND TS=(mouse OR mice OR rat OR rabbit OR dog OR swine OR pig OR sheep OR monkey OR fish OR flies OR"laboratory animal"OR"experiment animal"as the search formula.CiteSpace software was applied to visualize and analyze the authors,institutions,countries,keywords,and literature co-citations.RESULTS AND CONCLUSION:(1)After analyzing and screening,1 238 documents were included in the CNKI and WanFang databases,and 3 419 documents were included in the Web of Science Core Collection database.The overall number of articles issued since 1999 has been on an upward trend,with the highest research centrality in the United States,the highest number of articles issued in China.Moreover,the institution with the highest research centrality is the University of California.(2)After removing the keywords directly related to the article topics and synthesizing the co-occurrence frequency and centrality,"biomechanics,""Zuo Gui Wan,""alendronate,""breast cancer,"and"biochemical indicators"are in the core of the research field.(3)According to the analysis of co-citations,among the top 10 cited documents,5 of them are related to the treatment measures and clinical efficacy of postmenopausal osteoporosis.(4)Based on the analysis of keywords and co-cited literature,investigations on osteoblast formation and the mechanism of bone formation,the mechanism of metabolomics,the treatment of traditional Chinese medicine,signaling pathway,parathyroid hormone treatment,and sclerostin antibody treatment are the hot research topics now and the future research trends.

Objective To explore different construction models and experiences of clinical laboratory centers under exist-ing Integrated county healthservices entity,analyze and discuss the characteristics of various construction models,and provide ref-erences for the development of county-level clinical laboratory centers.Methods Based on the five factors of"man,machine,material,method,and environment"in Total Quality Management theory,an interview outline and questionnaire were designed for county-level clinical laboratory centers.Eleven county-level clinical laboratory centers across the country were investigated to analyze their construction models,investment returns,and other aspects.Literature reviews and case studies were also conducted to summarize the construction models and characteristics of county-level clinical laboratory centers.Results The construction models of county-level clinical laboratory centers are mainly divided into five types,each with distinct features.Through an analy-sis of the investment returns of these models unde rIntegrated county healthservices entity,it was found that a profit distribution ratio of approximately 5∶5 between county-level clinical laboratory centers and township health centers for referred samples is more conducive to maintaining the stability of sample sources.The construction model of county-level clinical laboratory centers is closely related to the medical service capacity of both county and township levels.Conclusion Counties with stronger county hospital capabilities tend to establish relatively independent regional laboratory centers based on the county hospital's laboratory department.Regardless of the model,the primary goal of county-level clinical laboratory centers should be to provide high-quality testing services coverage across the entire county.The key to the success of county-level clinical laboratory centers lies in motiva-ting grassroots personnel.