Objective: To investigate the effect of targeted silencing of DNA methyltransferase 3A(DNMT3A) on collagen deposition, proliferation and migration activity of mouse lung fibroblasts(PFs). Methods: In order to ensure the proliferation and migration activity of primary fibroblasts, the lung tissues of neonatal C57 suckling mice were taken, PFs were extracted after being sheared, and the morphology was observed and identified under the microscope. PFs cells were activated by 5 ng/ml TGF-β1for 24 h after cell attachment, and DNMT3A silencing model was constructed by small interfering RNA; The experiment was divided into control group, TGF-β1group, TGF-β1+ siRNA-NC group and TGF-β1+ siRNA-DNMT3A group. The protein expressions of DNMT3A, α-smooth muscle actin(α-SMA) and Collagen Ⅰ were detected by Western blot; Real time quantitative reverse transcription polymerase chain reaction(RT-qPCR) was used to detect the mRNA expression changes ofDNMT3A,α-SMAandCollagenⅠ. The proliferation ability of PFs was detected by CCK-8 and EdU staining; the migration ability of PFs was detected by scratch test and Transwell migration test. Results: Compared with the control group, TGF-β1induced the increase of DNMT3A in the activated PFs cell group(P<0.01), the protein and mRNA levels of fibrosis and proliferation related indicators α-SMA and Collagen Ⅰ also increased(allP<0.05), and the proliferation and migration ability of PFs increased(allP<0.000 1). Compared with the siRNA-NC group, the protein expression levels of DNMT3A(P<0.000 1) and related indicators α-SMA(P<0.01) and Collagen Ⅰ(P<0.01) significantly decreased in the DNMT3A silencing group by Western blot, and the mRNA levels ofDNMT3A,α-SMAandCollagenⅠby RT-qPCR also decreased(allP<0.001), and the proliferation(P<0.01) and migration ability(P<0.05) of PFs cells decreased compared with the control group. Conclusion Silencing DNMT3A can inhibit the deposition of collagen and the proliferation of PFs. DNMT3A can promote the proliferation and migration of PFs, and then promote the activation of PFs and the development of pulmonary fibrosis. This process may be regulated by DNA methylation modification.

The auxin/indole-3-acetic acid (Aux/IAA) gene family is an important regulator for plant growth hormone signaling, involved in plant growth, development, as well as response to environmental stresses. In the present study, we identified SmIAA7 which is potentially associated with Salvia miltiorrhiza leaf development through comparatively analyzed the transcriptome data from different pinnate leaves. SmIAA7 was successfully isolated from S. miltiorrhiza using the specific primers. Then subsequent bioinformatic analysis, prokaryotic expression and purification, subcellular localization, and induction expression analysis under auxin and abiotic stress were performed. The full-length of SmIAA7 contained an ORF of 684 bp encoding a protein of 227 amino acid with a molecular weight of 25.3 kD. Conserved domain analysis showed that SmIAA7 contains the conserved Aux_IAA domain (pfam02309). Sequence analysis and phylogenetic tree analysis results indicated SmIAA7 was phylogenetically close to IAA7 and IAA14 from other plants, suggesting SmIAA7 involved in plant growth and development as well as response to environmental stresses. The prokaryotic expression vector pET28a-SmIAA7 was constructed and SmIAA7 recombinant protein was successfully expressed in E. coli Rosetta (DE3) strain. Subcellular localization experiment demonstrated that SmIAA7 localized in the nucleus of plant cells. Real-time fluorescence quantitative PCR results showed that the expression level of SmIAA7 was upregulated in response to auxin. Drought, low temperature, and salt stress significantly increased the transcript level of SmIAA7 gene. This study lays a foundation for further elucidating the role of SmIAA7 in leaf development, signal transduction, and stress defense in S. miltiorrhiza.

ObjectiveTo investigate the perceived experience and acceptance of intrapartum ultrasound （IPUS） as a novel method for labor progress assessment among pregnant women. MethodsFrom February 2023 to December 2024， a total of 180 pregnant women admitted to the Labor Ward of Lingnan Hospital， the Third Affiliated Hospital of Sun Yat-sen University， who were planned for vaginal trial of labor ， were accessed for labor progress using IPUS and vaginal examination （VE） after the onset of labor and prior to the initiation n of labor analgesia. A self-designed questionnaire was used to investigate the women's perceived experiences with both examination methods and their acceptance of IPUS. The pain intensity associated with the examinations was evaluated using the visual analogue pain scale （VAS）. Differences in the women's experiences and pain intensity between the two labor progress assessment methods were compared. ResultsThe acceptance rate of IPUS was 96.67% （174/180）， with the remaining 6 cases undecided. Over 60% of the pregnant women reported IPUS assessment as comfortable and none of them felt discomfort， whereas 32.8% felt uncomfortable with VE （χ²=196.02， P＜0.001）. Nearly two-thirds of the pregnant women believed that VE would cause psychological distress， while none reported such effect with IPUS （χ²=261.52， P＜0.001）. Approximately 77.78% （140/180） of the pregnant women believed that IPUS could reduce their fear of vaginal delivery and enhance their confidence if it replaced VE. The VAS score for IPUS ［0 （0， 2）］ was significantly lower than that for VE ［4 （4， 6）］ （Z=-14.62， P＜0.001）. Further stratified analysis showed that over 90% （164/180） of the pregnant women found IPUS painless， with no moderate or severe pain reported， compared to 43.33% （78/180） experienced moderate or severe pain with VE （P＜0.001）. ConclusionAs a novel approach for labor progress assessment， IPUS not only alleviates the pain and discomfort associated with traditional VE and reduces the fear of childbirth but also enhances women's confidence in delivery， thereby achieving a high level of acceptance among parturient women in China.

Objective To analyze the epidemiological characteristics and spatiotemporal clustering of varicella in Lu'an City from 2005 to 2023, and to provide a scientific basis for optimizing varicella prevention and control strategies. Methods Data on varicella cases were collected through the Chinese Center for Disease Control and Prevention Information System. Descriptive epidemiology, temporal trend analysis, seasonal analysis, spatiotemporal clustering analysis, and spatial autocorrelation analysis were conducted using QGIS, JoinPoint, SaTScan and GeoDa software. Results The average annual reported incidence rate of varicella in Lu'an City from 2005 to 2023 was 34.55/100,000, showing a trend of initial increase followed by a decrease. The peak incidence occurred from October to January of the following year (RR=1.97, LLR=1743.95, P=0.001). Students aged 0 to 19 was the primary affected group. Spatiotemporal scan analysis revealed four types of spatiotemporal clusters, with the cluster in Jin'an District from October 2017 to December 2023 being particularly prominent (RR=2.87，LLR=1734.15，P<0.001). Spatial autocorrelation analysis indicated significant clustering of varicella cases in the main urban area (Moran's I=0.216，Z=4.786，P=0.003). Conclusion The incidence of varicella in Lu'an City exhibits distinct seasonal and spatial clustering, and schools and kindergartens in the main urban area are the key to varicella prevention and control. It is necessary to enhance the monitoring of disease outbreaks during peak periods and in key areas, and to increase the two-dose vaccination rate for varicella in areas with case aggregation and among key populations.

Aim To investigate the effect and role of neuronal restriction silencing factor(REST/NRSF)in epilepsy disorder.Methods Immunohistochemistry,immunofluorescence,Western blot and qPCR tech-niques were used to detect REST/NRSF expression levels in hippocampal tissues of mice induced by kainic acid and human brain tissue.Viral injections,EEG re-cordings and behavioral methods were used to test the effects on epileptic mice after knockdown and overex-pression of REST/NRSF in the hippocampal CA1 re-gion,respectively.Results The positive rate of REST/NRSF in the lesions of epileptic patients was significantly higher compared with that in the control group.The levels of REST/NRSF protein and mRNA in the CA1 region of the hippocampus of mice in the KA model group were significantly higher.Kv7.2 and Kv7.3 potassium channel mRNA expression levels were significantly down-regulated.Significant up-regu-lation of REST/NRSF expression levels was observed in mouse hippocampus after NMDA injection.Knock-down of REST/NRSF in the CA1 region of hippocam-pus significantly elevated the expression levels of Kv7.2 and Kv7.3 potassium channel mRNAs.The fre-quency of EEG spiking and sharp-wave issuance and epileptic seizure grade were significantly lower.Over-expression of REST/NRSF in the CA1 region of hippo-campus significantly reduced the mRNA expression lev-els of Kv7.2 and Kv7.3 potassium channels.The fre-quency of EEG spiking and sharp-wave issuance was significantly higher and epileptic symptoms were exac-erbated.Conclusion REST/NRSF in mouse hipp-ocampal brain regions is involved in epileptic disease development through transcriptional regulation of Kv7.2 and Kv7.3 potassium channels.

Objective:To deeply explore the experience of lactation mastitis women, so as to provide a basis for formulating reasonable support measures.Methods:This study was a qualitative study. From November to December 2022, purposive sampling was used to select 13 patients with lactation mastitis who visited the Breast Specialist Nursing Clinic of Dongfang Hospital, Beijing University of Chinese Medicine as the research subject. This study used semi-structured interviews to collect data, and applied the Colaizzi 7-step analysis method to analyze, summarize, and extract themes.Results:The experience of lactation mastitis patients were mainly summarized as self-emotional experience and interpersonal relationship experience. Self-emotional experiences included feelings of anxiety, stress, helplessness, and self-blame. Interpersonal relationship experiences consisted of feelings of helplessness, useless, decision-making difficulties, desire for help and understanding.Conclusions:Lactation mastitis patients face various negative psychological experiences and burdens. Medical and nursing staff, family caregivers, and society should jointly pay attention to the patients' feelings and provide targeted support measures.

Objective:To investigate the effect of deacylase Sirtuin 5 in the recovery of hematopoietic stem cells(HSCs)after treated by 5-FU in mouse.Methods:Flow cytometry was used to analyze the effect of SIRT5 deletion on the proportion of hematopoietic stem/progenitor cells(HSPCs)in bone marrow(BM),the proportion of T cells,B cells and myeloid cells(TBM)in peripheral blood(PB)and spleen,and the development of T cells in thymus.Mouse were treated with 5-FU to study the effect of SIRT5 deletion on the cell cycle,apoptosis and the proportion of HSPCs in BM.The effect of SIRT5 deletion on the proliferation of HSCs was analyzed by flow sorting in vitro.Results:SIRT5 deletion did not affect the development of T cells in thymus and the proportion of TBM cells in PB and spleen compared with wild type mice.SIRT5 deletion increased proportion of HSPCs in BM.After 5-FU treatment,the proportion of HSCs in SIRT5 deletion mice was significant decreased(P＜0.05),the HSPC in SIRT5 deletion mice was activated from G0 to G1 phase(P＜0.05),and the proportion of early apoptosis increased(P＜0.05).By monoclonal culture in vitro,the ability of HSCs to form clones in SIRT5 deletion mice was decreased significantly(P＜0.05).Conclusion:SIRT5 deletion lead to a decreased the ability of HSCs to clone in vitro.SIRT5 deletion is not conducive to the recovery of HSPCs injury in mice under hematopoietic stress.

Objective:To investigate the relative expression level and clinical significance of LINC00475 in serum of patients with multiple myeloma(MM).Methods:The expression of LINC00475 in serum of 108 MM patients and five MM cell lines including RPMI 8226,NCI-H929,U266,OPM2 and CAG were detected by real-time fluorescence quantitative PCR.The diagnostic value of LINC00475 in MM was evaluated by receiver operating characteristic(ROC)curve analysis.The correlation of LINC00475 with patients'characteristics was analyzed.Results:Compared with control groups,the expression of LINC00475 was up-regulated in serum of MM patients and MM cell lines(all P＜0.05).ROC curve analysis showed that the optimal cut-off value of LINC00475 was 262.4,the area under curve(AUC)was 0.924(95％CI:0.884-0.964),and sensitivity and specificity was 83.3％and 91.7％,respectively,which indicated that LINC00475 had good evaluation value in MM patients.Compared with low-LINC00475 expression group,patients in high-LINC00475 expression group had higher levels of β2-microglobulin(β2-MG)and Cystatin C(Cys-C)but lower albumin(ALB)(all P＜0.05).Compared with MM patients with International Staging System(ISS)stage I,the expression level of LINC00475 was significantly higher in patients with stage Ⅱ and Ⅲ(both P＜0.05).Conclusion:LINC00475 is helpful to distinguish MM patients from healthy adults,which is correlated with the prognostic indicators such as β2-MG,ALB,and ISS stage.

Objective To retrieve,evaluate and summarize the best evidence on the management of ocular complications in intensive care patients ventilated in prone position,and to provide references for clinical practice.Methods Evidence on management of ocular complications in intensive care patients ventilated in prone position was systemically retrieved in the guideline websites,professional association websites and databases,such as the BMJ Best Practice,UpToDate,Cochrane Library,Joanna Briggs Institute,PubMed,Web of Science,Science Direct,Embase,CNKI,Wanfang and other databases,including guidelines,clinical decisions,evidence summaries,expert consensuses,group standards,systematic reviews and scoping reviews,published from January 2013 to June 2023.The guidelines were individually evaluated by 4 researchers,and the remaining literature was individually evaluated by 2 researchers.The literature that met the criteria was extracted and graded.Results A total of 15 articles were involved,including 6 guidelines,2 clinical decisions,2 evidence summaries,3 systematic reviews,1 scoping review and 1 group standard.Finally,6 evidence topics and 16 pieces of best evidence were formed,including organization and training,risk identification,eye assessment,eye protection,position management and regular observation.Conclusion This study summarized the best evidence on the management of ocular complications in intensive care patients ventilated in prone position.In the application,the best evidence should be selected according to the clinical situation,so as to reduce the incidence of ocular complications and improve the quality of life of patients.

Objective:To preliminarily investigate the applicability of a poliovirus pseudovirus-based neutralization assay in evaluating the immunogenicity of recombinant poliovirus vaccines and their in vivo potency in rats. Methods:Serum samples from rats immunized with recombinant poliovirus vaccines were tested using both the pseudovirus neutralization assay and the live-virus neutralization assay with Sabin strain. The consistency and correlation of the two methods were analyzed using the Kappa test and Spearman′s rank correlation.Results:For the neutralizing antibodies against typeⅠ, Ⅱ, and Ⅲ polioviruses, the Kappa values for consistency analysis of the two methods were 0.914, 1.000, and 0.751, respectively ( P<0.001), and the correlation coefficients ( R values) were 0.833, 0.927, and 0.859, respectively ( P<0.001). Conclusions:The test results of the two methods are consistent and show a good correlation, indicating that the pseudovirus neutralization assay can be applied to evaluating the immunogenicity of poliovirus vaccines and also can be used in rat potency tests.