OBJECTIVE To investigate the effects of total flavonoids from Carthamus tinctorius L. （TFCTL） on hepatic stellate cell （HSC） activation based on the microRNA （miRNA）-204/NUAK family SNF1-like kinase 1 （NUAK1）/Hippo signaling axis， thereby elucidating the potential mechanism underlying their antifibrotic effects. METHODS The HSC-T6 cells were divided into control group， model group， TFCTL low-concentration group （20 μg/mL）， TFCTL medium-concentration group （40 μg/mL）， and TFCTL high-concentration group （60 μg/mL）. Except for control group， the remaining groups were treated with 5 ng/mL of transforming growth factor-β to induce the activation of hepatic stellate cells， followed by the addition of corresponding drug solutions/culture medium and incubation for 24 hours. Cell apoptosis was assessed， the expression levels of α-smooth muscle actin （α-SMA）， type Ⅰ collagen （Collagen Ⅰ） and proteins associated with the Hippo/Yes-associated protein （YAP） pathway [YAP， large tumor suppressor kinase 1 （LATS1）， and mammalian STE20-like kinase 1 （MST1）] were detected. Additionally， cell transfection was used to investigate the activity of the miRNA-204/NUAK1/Hippo signaling axis at both the genetic and protein levels. RESULTS After intervention with TFCTL， the apoptosis rate of HSC-T6 cells and the protein expressions of MST1 （except for the TFCTL high-concentration group） and LATS1 were significantly increased （P＜0.05）， while the protein expressions of α-SMA， CollagenⅠ， and YAP （except for the TFCTL medium-concentration group） were significantly decreased （P＜0.05）. Further results from cell transfection experiments revealed that after transfection with miRNA-204 mimics， the mRNA it’s protein expressions of α-SMA， CollagenⅠ， NUAK1， and YAP in HSC-T6 cells were significantly decreased （P＜0.05）， while the mRNA and protein expressions of LATS1 and the mRNA expression of MST1 were significantly increased （P＜0.05）. Conversely， the results were opposite following transfection with miRNA-204 inhibitors. CONCLUSIONS TFCTL can exert anti-hepatic fibrosis effects by up-regulating the expression of miRNA-204， thereby down- regulating the expressions of NUAK1， inactivating the Hippo/YAP pathway， which in turn suppresses the activation of HSC and promotes their apoptosis.

Objective: To analyze the changes in classroom lighting environment of schools in Suzhou and their impact on refractive progression among primary and secondary school students, so as to provide the basis for accurate provention and control of myopia. Methods: A baseline investigation was conducted in October 2022 by using a stratified cluster random sampling method to recruit primary and secondary school students from Suzhou. A follow up visit was performed in October 2023. A total of 12 302 students and 360 classrooms that participated in both surveys were included analysis. The visual acuity progression over one year and classroom lighting conditions were assessed. Group comparisons were performed by using the Wilcoxon or Kruskal-Wallis rank-sum, and Chi-square tests. Multivariate Logistic regression was employed to identify the major factors influencing refractive changes. Results: The compliance rate of average illuminance on classroom blackboard surface increased from 72.22% to 75.28%, while the compliance rate of average illuminance on desks decreased from 89.44% to 87.22%, the overall myopia rate among students rose from 59.63% to 66.99% from 2022 to 2023. The average annual progression of equivalent spherical power(SE) in the right eye of students was -0.25(-0.75,0.06)D. Significant statistical differences were observed in the annual mean changes across different school levels, regions, baseline refractive statuses, and classroom lighting environment change groups ( Z/H =316.59, -8.27, 38.80 , 51.01, all P <0.05). Multivariate Logistic regression analysis showed that pre myopia, low myopia, junior high school, senior high school, vocational high school, and improved classroom lighting environment were protective factors of reducing the risk of rapid progression in refractive error ( OR =0.58, 0.69, 0.81, 0.50, 0.28, 0.82, all P <0.05). Conversely, female students and rural students had higher risks of rapid myopia progression ( OR =1.09, 1.42, both P <0.05). Conclusions Over one year follow up, the complance rate of classroom lighting indicators in Suzhou remaines stable, while students refractive status shows a trend toward myopia. Improving classroom lighting environment can reduce the risk of rapid myopia progression.

[摘 要] 免疫检查点抑制剂（ICI）已广泛应用于多种实体肿瘤的治疗，循环T淋巴细胞亚群精细分型因其具有作为疗效预测标志物的前景而备受关注。目前常用的ICI疗效预测标志物多依赖于肿瘤组织样本，存在取材困难且难以实现动态监测的局限性。相比之下，循环T淋巴细胞亚群精细分型不仅能够反映T细胞的功能状态以预测ICI治疗反应，还因其取样便捷、微创安全的优势，成为一种可行的动态检测手段。基于功能状态，循环T淋巴细胞亚群精细分型主要分为活化、增殖、衰老及耗竭等表型。活化表型和增殖表型通常反映T细胞的活化、增殖和功能激活状态；而衰老表型与耗竭表型则反映T细胞储备下降，增殖及生存能力减弱、存活期缩短，效应功能降低或失能的状态，这些功能状态与ICI的疗效密切相关。本文系统综述基于外周血T细胞功能状态的精细分型在ICI治疗疗效预测中的最新研究进展，探讨不同功能状态的预测价值，分析其作为潜在预测工具的临床应用前景与现实困境，为后续技术优化与临床转化提供参考。

Cataract is the world's leading cause of blindness, and surgery is the most effective treatment for cataract. With the development of femtosecond laser technology and ophthalmic surgical equipment, the application of femtosecond laser systems in cataract surgery is becoming increasingly widespread. It can be used in cataract surgery for corneal incisions, anterior capsulotomy, lens fragmentation, arcuate incisions and other key operations. Compared to traditional surgery, femtosecond laser assisted cataract surgery(FLACS)offers significant advantages in precision, safety and postoperative visual outcomes. Its clinical benefits have garnered growing recognition among ophthalmologists. However, the key technologies and high-precision equipment for FLACS remain predominantly controlled by Western countries. In China, the research in this field began later. This article reviews the technological advancements in FLACS, with a focus on femtosecond laser technology, optical coherence tomography(OCT), artificial intelligence, and clinical application progress. The objective is to provide theoretical foundations and practical insights for the development of ophthalmic medical technology in China.

Objective To explore the mechanism of excessive mechanical stress regulated ferroptosis induced by Piezo1 channel in mouse chondrocytes.Methods The experiment was performed on mouse ATDC5 chondrocytes.siRNA-Piezo1 interference plasmid and Piezo1 overexpression plasmid were used to transfect chondrocytes,and mechanical stress stimulation was given.The control group,the mechanical stress stimulation group(MS group),the MS+siRNA-Piezo1 group(MS+sh group)and the MS+Piezo1 overexpression group(MS+OV group)were constructed,respectively.The cell viability,Fe2+,ROS levels,the expression of ferroptosis-related proteins SLC7A11 and GPX4,and the expression of Collagen Ⅱ,MMP-13,Aggrecan and p53 proteins were detected in each group.Results Compared with the control group,the cell viability was decreased in the MS group(P＜0.05).Levels of Fe2+,reactive oxygen species(ROS)and malondialdehyde(MDA)were increased(P＜0.05).Levels of reduced glutathione(GSH)and superoxide dismutase(SOD)were decreased(P＜0.05),and the mitochondrial ridge was decreased detected by transmission electron microscopy.Protein levels of SLC7A11,GPX4,Collagen Ⅱ and Aggrecan were decreased(P＜0.05),while protein levels of p53 and MMP-13 were increased(P＜0.05).Compared with the MS group,Fe2+,ROS and MDA levels were decreased in the MS+sh group(P＜0.05),GSH and SOD levels were increased(P＜0.05),and protein levels of SLC7A11,Collagen Ⅱ,GPX4 and Aggrecan were increased(P＜0.05).The protein levels of MMP-13 and p53 were decreased(P＜0.05).Compared with the MS group,cell viability was decreased(P＜0.05),Fe2+,ROS and MDA levels were increased(P＜0.05),GSH and SOD levels were decreased(P＜0.05),and protein levels of SLC7A11,Collagen Ⅱ,GPX4 and Aggrecan were decreased in the MS+OV group(P＜0.05).Levels of MMP-13 and p53 protein were increased(P＜0.05).Conclusion Excessive mechanical stress can induce chondrocyte ferroptosis and promote extracellular matrix degradation via Piezo1 channel protein.

BACKGROUND:Early diagnosis of pulmonary fibrosis is the foundation for timely antifibrotic drug therapy.Therefore,exploring and discovering ideal biomarkers that can be effectively used for the early diagnosis of pulmonary fibrosis is crucial for the treatment of the disease.OBJECTIVE:To conduct an in-depth analysis of key autophagy-related genes involved in the process of pulmonary fibrosis by means of bioinformatics and machine learning techniques,in order to investigate whether autophagy-related core genes of pulmonary fibrosis can be used as reliable biomarkers in the assessment of the progression of pulmonary fibrosis.METHODS:Two datasets of pulmonary fibrosis,GSE24206 and GSE110147,were downloaded from the Gene Expression Omnibus(GEO)database(a public database developed and maintained by the U.S.National Center for Biotechnology Information to store and share bioinformatics data),and the gene expression matrices of these two datasets were normalized by using the"limma"package in R software.The autophagy-related genes were extracted from GeneCards database(a database created by the U.S.National Center for Biotechnology Information,which automatically integrates gene-centric data from about 200 Web sources,including genomic,transcriptomic,proteomic,genetic,clinical,and functional information).Differential gene analysis was performed on the pulmonary fibrosis dataset,and the common genes were extracted by cross-comparing the differential genes with the autophagy genes,so as to identify autophagy genes that may play a role in the process of pulmonary fibrosis.The intersecting genes were analyzed for functional enrichment and cellular immune infiltration by gene ontology and Kyoto Encyclopedia of Genes and Genomes.Core genes of pulmonary fibrosis associated with autophagy were screened by protein-protein interactions and machine learning,and core genes were subjected to the enrichment analysis.Diagnostic models were constructed from the identified core genes.Calibration curves were used to assess the predictive ability of the line graph model.An external dataset,GSE21369,was used to perform a receiver operating characteristic curve analysis to validate the expression profiles of pulmonary fibrosis genes associated with autophagy,as well as to predict Chinese herbs associated with the genes IL6 and COL1A2 via the Coremine database.Finally,human embryonic lung fibroblasts were cultured and modelled by transforming growth factor-β1 treatment,and the relative expression of genes in the model cells was verified using qRT-PCR.RESULTS AND CONCLUSION:(1)A total of 51 pulmonary fibrosis differential genes and 25 genes intersecting with autophagy genes were obtained.Gene ontology analysis showed that the 25 intersecting genes were related to extracellular matrix tissue,collagen metabolism,collagen pro-fibroblasts,and growth factor binding,etc.The results of Kyoto Encyclopedia of Genes and Genomes enrichment analysis indicated that they were mainly related to the Phosphatidylinositol 3-kinase/protein kinase B signaling pathway and the signaling pathway of the extracellular matrix-receptor interactions.(2)Immunoinfiltration analysis revealed that the expression of activated memory CD4+T cells,M0 macrophages,and resting dendritic cells was significantly elevated in the pulmonary fibrosis group(P＜0.05),showing a strong correlation.(3)Two autophagy signature genes involved in the progression of pulmonary fibrosis were identified:COL1A2 and IL6.The column-line diagram model showed that the two core genes predicted the onset of pulmonary fibrosis more accurately,and the receiver operating characteristic curve analysis showed that the two characteristic genes had diagnostic significance.COL1A2 and IL6 were related to the cell-cycle pathway,mitogen-activated protein kinase signaling pathway,Janus kinase-signal transduction and activator of transcription signaling pathway and cytokine-cytokine receptor interactions.A total of 20 Chinese herbs were predicted to be related to COL1A2 and IL6 genes,and their efficacies were mainly to clear away heat and detoxify toxins and to invigorate blood and move qi.COL1A2 and IL6 were verified to be highly expressed in pulmonary fibrosis.To conclude,COL1A2 and IL6 may be potential diagnostic biomarkers for pulmonary fibrosis,but its specificity to pulmonary fibrosis needs to be further investigated.

Objective To develop nanobodies with broad-spectrum reactivity,specificity,and high sensitivity that can be used for detecting multiple subtypes of influenza A virus,and to establish a nanobody-based enzyme-linked immunosorbent assay(ELISA)method.Methods Gene sequences of twelve nanobodies against influenza A virus were retrieved from the National Center for Biotechnology Information(NCBI)and nanobody databases.The nanoantibodies were prepared using molecular biological techniques including gene synthesis and recombinant expression.The binding activity,specificity,sensitivity,and affinity of these nanobodies were determined by ELISA screening and Gator affinity analysis.A double-antibody sandwich ELISA assay was established by combining the selected nanobody with a traditional mouse monoclonal antibody.Results Twelve nanobodies were expressed and purified.Two nanobodies capable of binding to multiple subtypes of influenza virus including H1,H3,H5,H7,and H9 were obtained and designated as VHH54 and KV108.Both nanobodies showed no cross-reactivity with other respiratory virus antigens.Furthermore,the KV108 nanobody exhibited the highest binding affinity,with a dissociation constant of 5.94×10-9mol/L for the influenza virus nucleoprotein(NP),and the lowest detection concentration for the NP antigen reached 0.00064 μg/mL.The double-antibody sandwich ELISA,using a combination of KV108 and a mouse monoclonal antibody,could sensitively detect the five common subtypes of influenza A virus(H1N1,H3N2,H5N1,H7N9,and H9N2).The lowest detection limit reached 110-403 PFU/mL,which was higher than that of the commercial colloidal gold kitfor influenza virus detection.Conclusion This study has identified a nanobody KV108,which is capable of binding to multiple subtypes of influenza virus,and established a nanobody-based ELISA method that can detect multiple subtypes of influenza A virus.This study can facilitate the development of nanobody-based influenza detection technologies.

Objective:To explore the application of contrast-enhanced ultrasound in the preoperative evaluation of carotid body tumor (CBT).Methods:The clinical data of 13 CBT patients undergoing contrast enhanced ultrasound test and surgical treatment at Peking Union Medical College Hospital from Nov 2017 to Aug 2021 was retrospectively analyzed.Results:Among the 13 patients, 7 patients had bilateral lesions. 18 tumors were identified by contrast enhanced ultrasound, which showed rich blood supply, with marked enhancement in 13 tumors and moderate enhancement in 5 tumors. The origins of the arterial supply for tumors were identified by contrast enhanced ultrasound. Time-intensity curve analysis showed that the tumors had enhancement characteristics of fast wash in and slow wash out. The mean contrast wash in time was (3.33±1.40) s, the mean peak intensity was (10.41±1.74) dB, and the mean wash out time was (56.47±22.28) s. A total of 13 cases underwent successful surgical removal. Five cases of external carotid artery ligation and 2 cases of internal carotid artery reconstruction were performed during surgery. Postoperative transient neurological injury occurred in 5 cases. There were no cases of cerebral infarction or death in the perioperative period. Mean postoperative follow-up was 14.31 months. Five cases of neurological injury had satisfactory recovery and no other adverse events occurred.Conclusions:Contrast enhanced ultrasound is an effective method of preoperative imaging assessment for CBT, which helps the surgical planning and preoperative preparation.

Objective To establish human colorectal cancer(CRC)organoids and to evaluate the efficacy of chem-otherapy drugs.Methods Patient-derived CRC cells were cultured to form organoids.The CRC organoids and origi-nal tissues were stained with molecular markers of CRC immunohishtochemically.CRC organoids were used to test drug sensitivity and different concentrations of chemotherapy drugs 5-fluorouracil,oxaliplatin and irinotecan were given respectively;Organoid activity before and after drug treatment was measured by 3D cell viability assay.Results The patient-derived organoids(PDO)from 5 CRC tissues were successfully established.The expression of CK20,Ki67 and Villin proteins was similar in organoids and in original tumor.The organoids retained histologcial features similar to those of the original tumors.Different PDO showed differential sensitivity to different chemothera-py drugs.Conclusions CRC-PDO can dispaly their different sensitivities to different chemotherapy drugs,and could provide valuble reference for personalized treatment for CRC patients.

The malignancy of pancreatic cancer is high. Radical resection is the main treatment method for non-metastatic pancreatic ductal adenocarcinoma. Neoadjuvant therapy can improve the surgical resection rate of borderline resectable pancreatic ductal adenocarcinoma (BR-PDAC) and prolong the survival of patients. In recent years, a large number of studies have been conducted on neoadjuvant therapy for pancreatic cancer at home and abroad, including radiotherapy, chemotherapy, immunotherapy and combination therapy. This article mainly reviews the current research status of neoadjuvant therapy for BR-PDAC.