ObjectiveTo investigate the role of Ras-GTPase-activating protein SH3 domain-binding protein 2 （G3BP2） in regulating the activation， proliferation， and migration of hepatic stellate cells （HSCs）. MethodsThe mouse HSCs （JS-1 cell line） were treated with 5 μg/L transforming growth factor-beta 1（TGF-β1） for 24 hours to establish an HSC activation and proliferation model. A G3BP2 knockdown system was constructed using siRNA interference technology. The experiment was divided into four groups： Control， TGF-β1 treatment， TGF-β1+si-NC， and TGF-β1+ G3BP2-siRNA. The expression levels of key fibrosis indicators， including type I collagen （Collagen I）， α-smooth muscle actin （α-SMA）， and G3BP2， were detected by Western blot and RT-qPCR. Cell proliferation activity was assessed using the CCK-8 proliferation assay kit and EdU fluorescence labeling technology. Cell migration ability was analyzed by scratch wound healing assay and Transwell migration assay. The formation level of stress granules was quantified by immunofluorescence microscopy to investigate the effects of G3BP2 on stress granule formation in activated HSCs. ResultsStimulation with TGF-β1 upregulated the expression of G3BP2 in JS-1 cells （RT-qPCR： P0.000 1； Western blot： P0.000 1）， while a downward trend in its expression was observed in the G3BP2‑silenced group （RT-qPCR： P0.01； Western blot： P0.000 1）. Compared with the control group， the TGF-β1 group exhibited increased protein expression levels of α-SMA and Collagen I （RT-qPCR： both P0.01； Western blot： P0.01 and P0.05， respectively）， concomitant with an increased number of stress granules and enhanced cell proliferation and migration capacity （all P0.001）. The experimental results demonstrated that G3BP2 knockout effectively reversed the aforementioned phenotypes， with the G3BP2-silenced group showing reduced expression of fibrotic markers （all P0.01）， decreased stress granule formation （P0.01）， and reduced cell proliferation and migration capacity （all P0.05）， compared to the negative control group. ConclusionG3BP2 enhances the activation， proliferation， and migration of HSCs by promoting the formation of stress granules， thereby accelerating the pathological progression of liver fibrosis. This suggests that stress granules may serve as important regulators in controlling the activation， proliferation， and migration of HSCs.

Based on the view that wind pathogen runs through the entire course of the occurrence and development of nephrotic syndrome， it is proposed that the core pathogenesis of refractory nephrotic syndrome （RNS） in children involves imbalance of the lung， spleen， and kidney， invasion of wind pathogen， and stagnation of turbidity and stasis， and that directing the turbid downward with wind medicinals is the key therapeutic strategy. In clinical practice， according to the distinct pathogenic characteristics of three patterns， including defensive qi insecurity of the lung， yang deficiency with loss of regulation， and binding of turbidity and stasis， three corresponding methods are applied， which are diffusing the lung to consolidate defensive qi， elevating yang to supplement the spleen and kidney， and activating blood to unblock turbidity and stasis. For the pattern of defensive qi insecurity of the lung， wind medicinals are used to penetrate and disperse external pathogens， such as fangfeng （Radix Saposhnikoviae） and mahuang （Herba Ephedrae） for diffusing the lung， dispersing wind， and consolidating defensive qi. For yang deficiency with loss of regulation pattern， wind medicinals are employed to elevate yang and assist transformation， such as shengma （Rhizoma Cimicifugae） and gegen （Radix Puerariae Lobatae） for raising clear yang， warming and activating the spleen and kidney. For binding of turbidity and stasis pattern， wind medicinals with scurrying and collateral‑unblocking properties are used， such as jixueteng （Caulis Spatholobi）， luoshiteng （Caulis Trachelospermi）， and chuanshanlong （Rhizoma Dioscoreae Nipponicae） for dispelling wind， activating blood， resolving stasis， and directing turbidity downward. In all these methods， the core principle is to use wind medicinals to regulate qi movement， open and unblock the sweat pores， and separately eliminate turbid pathogens， thereby providing a theoretical and practical basis for the traditional Chinese medicine diagnosis and treatment of RNS in children.

Based on the view that wind pathogen runs through the entire course of the occurrence and development of nephrotic syndrome， it is proposed that the core pathogenesis of refractory nephrotic syndrome （RNS） in children involves imbalance of the lung， spleen， and kidney， invasion of wind pathogen， and stagnation of turbidity and stasis， and that directing the turbid downward with wind medicinals is the key therapeutic strategy. In clinical practice， according to the distinct pathogenic characteristics of three patterns， including defensive qi insecurity of the lung， yang deficiency with loss of regulation， and binding of turbidity and stasis， three corresponding methods are applied， which are diffusing the lung to consolidate defensive qi， elevating yang to supplement the spleen and kidney， and activating blood to unblock turbidity and stasis. For the pattern of defensive qi insecurity of the lung， wind medicinals are used to penetrate and disperse external pathogens， such as fangfeng （Radix Saposhnikoviae） and mahuang （Herba Ephedrae） for diffusing the lung， dispersing wind， and consolidating defensive qi. For yang deficiency with loss of regulation pattern， wind medicinals are employed to elevate yang and assist transformation， such as shengma （Rhizoma Cimicifugae） and gegen （Radix Puerariae Lobatae） for raising clear yang， warming and activating the spleen and kidney. For binding of turbidity and stasis pattern， wind medicinals with scurrying and collateral‑unblocking properties are used， such as jixueteng （Caulis Spatholobi）， luoshiteng （Caulis Trachelospermi）， and chuanshanlong （Rhizoma Dioscoreae Nipponicae） for dispelling wind， activating blood， resolving stasis， and directing turbidity downward. In all these methods， the core principle is to use wind medicinals to regulate qi movement， open and unblock the sweat pores， and separately eliminate turbid pathogens， thereby providing a theoretical and practical basis for the traditional Chinese medicine diagnosis and treatment of RNS in children.

ObjectiveTo observe the clinical efficacy and safety of Tianmeng Lingjiao Formula（天礞羚角方， TLF） as an adjunctive therapy for paroxysmal atrial fibrillation （PAF） with phlegm-heat harassing the interior syndrome. MethodsNinety patients with PAF and phlegm-heat harassing the interior syndrome were enrolled and randomly assigned in a 2∶1 ratio to test group （60 cases） and control group （30 cases）. The control group received conventional western medicine， while the test group received TLF in addition to the conventional treatment， with one dose administered daily. The treatment course for both groups was 4 weeks. The efficacy of traditional Chinese medicine （TCM） syndrome and the rate of atrial fibrillation （AF） conversion were compared between groups. Assessments included AF burden， frequency and duration of AF episodes， TCM syndrome scores， and European Heart Rhythm Association （EHRA） AF symptom classification before and after treatment. Safety indicators such as blood routine， urine routine， stool routine， liver function， kidney function， and electrolytes were also monitored. Adverse events were recorded. ResultsA total of 82 patients completed the study （54 in the test group and 28 in the control group）. After treatment， AF burden， the frequency and duration of AF episodes， and TCM syndrome scores were significantly reduced in both groups， and the test group showed lower indicators than the control group （P＜0.01 or P＜0.05）. The AF conversion rate was 25.93% （14/54） in the test group， significantly higher than 3.57% （1/28） in the control group （P＜0.05）. The total effective rate for TCM syndrome efficacy was 83.33% （45/54） in the test group， which was significantly higher than 67.86% （19/28） in the control group （P＜0.05）. After treatment， the EHRA atrial fibrillation symptom severity in the test group showed significant improvement， and the improvement in atrial fibrillation symptoms was superior to that in the control group （P＜0.01or P＜0.05）. No significant abnormalities were observed in safety indicators， and no adverse events occurred during the trial. ConclusionTLF as an adjunctive therapy can reduce AF burden， enhance the conversion rate of AF， decrease the frequency and duration of episodes， and improve clinical symptoms in patients with paroxysmal atrial fibrillation presenting with phlegm-heat harassing the interior syndrome， with a favorable safety profile.

Objective: To explore the effect of YTH domain family protein 1(YTHDF1) on the activation, proliferation and migration of hepatic stellate cells(HSCs) by regulating mitochondrial fission mediated by mitochondrial fission protein 1(Fis1). Methods: The mouse hepatic stellate cell line JS-1 was treated with 5 ng/ml TGF-β1 for 24 h to induce its activation and proliferation, andYTHDF1-siRNA was used to construct aYTHDF1silencing model.The experiment was divided into Control group, TGF-β1 group, TGF-β1+si-NC group and TGF-β1+si-YTHDF1 group.Expression changes ofYTHDF1,Fis1and key indicators of fibrosis, type Ⅰ collagen(CollagenⅠ) and α-smooth muscle actin(α-SMA) were detected through reverse transcription quantitative polymerase chain reaction(RT-qPCR) and Western blot; CCK-8 was used to detect cell proliferation ability; Transwell migration assay and cell scratch assay were used to detect cell migration ability; immunofluorescence staining experiment was used to detect the effect ofYTHDF1onFis1-mediated mitochondrial fission; finally, JC-1 staining was used to experimentally detect the effect ofYTHDF1on mitochondrial membrane potential. Results: Compared with the Control group, RT-qPCR and Western blot experimental results showed that the expression ofYTHDF1andFis1increased in the TGF-β1 group(P<0.05,P<0.01;P<0.000 1), as well as the fibrosis markersCollagenⅠand the expression level of α-SMA increased(P<0.01;P<0.001,P<0.000 1); while adding CCK-8, the experimental results showed that the proliferation ability of HSCs in the TGF-β1 group was enhanced(P<0.000 1); Transwell experimental results showed that the migration ability of HSCs in the TGF-β1 group was enhanced(P<0.01); the cell scratch experiment results showed that the migration ability of HSCs in the TGF-β1 group was enhanced(P<0.000 1); the immunofluorescence experiment results showed that the TGF-β1 group Mito-Tracker Red staining andFis1co-localization signal increased(P<0.05); JC-1 staining experiment results showed that the mitochondrial membrane potential increased in the TGF-β1 group(P<0.01). Compared with the TGF-β1+si-NC group, RT-qPCR and Western blot experimental results showed that the expression ofYTHDF1andFis1in the TGF-β1+si-YTHDF1 group was reduced(P<0.01;P<0.001), and fibrosis markers the levels ofCollagenⅠandα-SMAwere reduced(P<0.01;P<0.001,P<0.01).CCK-8 experimental results showed that the proliferation ability of HSCs in the TGF-β1+si-YTHDF1 group was weakened(P<0.000 1); Transwell experiment results showed that the migration ability of HSCs in the TGF-β1+si-YTHDF1 group was weakened(P<0.001); cell scratch experiment results showed that the migration ability of HSCs in the TGF-β1+si-YTHDF1 group was weakened(P<0.000 1); immunofluorescence experiment results showed that the Mito-Tracker Red staining andFis1co-localization signal decreased in the TGF-β1+si-YTHDF1 group(P<0.01); JC-1 staining experiment results showed that mitochondrial membrane potential decreased in the TGF-β1+si-YTHDF1 group(P<0.05). Conclusion YTHDF1promotes the activation, proliferation and migration capabilities of HSCs by positively regulatingFis1-mediated mitochondrial fission. This suggests thatYTHDF1may be a key gene involved in regulating the activation, proliferation and migration of HSCs.

BACKGROUND:Multiple studies have confirmed that mitogen-activated protein kinase(MAPK)signaling pathway is involved in cell proliferation,and microRNA(miR)is involved in the occurrence and development of hypertrophic scars.Therefore,the role of miR-27a-3p and MAPK signaling pathways in pathological scar formation has been further explored. OBJECTIVE:To explore the effect of miR-27a-3p on the proliferation of human hypertrophic scar fibroblasts through the MAPK signaling pathway. METHODS:The primary fibroblasts were isolated and collected from the skin samples.The primary fibroblasts were observed by inverted microscope and verified by immunofluorescence.The relative expression level of miR-27a-3p in tissues was detected by qRT-PCR.The target genes of hsa-miR-27a-3p were predicted using the database,and then the predicted target genes were enriched by gene ontology function analysis and biological pathway enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes.There were seven groups:blank control,negative control,miR-27a-3p mimic,miR-27a-3p inhibitor,miR-27a-3p mimic+p38 MAPK inhibitor,miR-27a-3p mimic+extracellular regulated protein kinase inhibitor,miR-27a-3p mimic+c-Jun N-terminal kinase inhibitor.Western blot was used to detect the levels of extracellular regulated protein kinase,c-Jun N-terminal kinase inhibitor.and p38 kinase and their phosphorylation levels.Cell counting kit-8 and EdU were used to detect cell proliferation. RESULTS AND CONCLUSION:Compared with normal skin fibroblasts,hypertrophic scar fibroblasts had stronger proliferative activity(P<0.05)and faster proliferation level(P<0.001).Compared with normal skin,miR-27a-3p was highly expressed in hypertrophic scars(P<0.001).Compared with the negative control group,overexpression of miR-27a-3p could promote cell proliferation activity(P<0.001)and proliferation levels(P<0.001).Compared with the negative control group,knockdown of miR-27a-3p could inhibit the proliferation activity(P<0.05)and proliferation levels(P<0.001).Compared with the negative control group,overexpression of miR-27a-3p promoted the phosphorylated levels of extracellular regulated protein kinase,c-Jun N-terminal kinase,and p38 mitogen-activated protein kinase(P<0.05).Compared with the negative control group,knockdown of miR-27a-3p inhibited the phosphorylated levels of extracellular regulated protein kinase,c-Jun N-terminal kinase,and p38 MAPK(P<0.05).Compared with the miR-27a-3p mimic group,specific inhibitors of extracellular regulated protein kinase,c-Jun N-terminal kinase,and p38 MAPK reversed the effects of miR-27a-3p on the proliferative activity(P<0.01)and proliferation level(P<0.001)of fibroblasts.To conclude,these results suggest that miR-27a-3p promotes the proliferation of human hypertrophic scar fibroblasts by activating the MAPK signaling pathway.

Objective To investigate the situation and influencing factors of preserved ratio impaired spirometry (PRISm) among dust-exposed workers in a wooden furniture manufacturing enterprise. Methods A total of 562 dust-exposed workers from a wooden furniture enterprise in Guangdong Province were selected as the study subjects using the convenience sampling method. The result of health-status questionnaire and occupational medical examinations among the participants were studied, and the influencing factors of PRISm were studied using the binary logistic regression analysis method. Results The detection rate of PRISm was 22.1% (124/562) among the study subjects. Binary logistic regression result showed that male workers had a higher risk of PRISm than female workers (P<0.01). Current smokers had a higher risk of PRISm than non-smokers (P<0.05). Workers with longer duration of exposure to occupational hazards had a higher risk of PRISm (P<0.05), those with higher grade of small-airway dysfunction had a higher risk of PRISm (P<0.01). In terms of body mass index, overweight or obese workers showed a higher detection rate of PRISm than those with normal weight (P<0.01). Greater amount of smoking pack per year had a higher risk of PRISm (P<0.01). Conclusion Dust-exposed workers in the wooden furniture manufacturing industry show a relatively high detection rate of PRISm. Male, current smoker, longer duration of exposure to occupational hazards, small airway dysfunction, overweight or obese, and smoking pack per year are influencing factors of PRISm among the dust-exposed workers in the wooden furniture manufacturing industry.

Objective To develop a tiered training framework for operating room nurses in tertiary hospitals,provi-ding guidance for corresponding training curriculum development and clinical competency evaluation.Methods Delphi method was used to conduct two rounds of expert consultation on the preliminary training indicators,estab-lishing a comprehensive tiered training framework for operating room nurses.Results After two rounds of expert consultation,a tiered training framework for operating room nurses was constructed,which includes 6 primary indi-cators,19 secondary indicators and 62 tertiary indicators.The response rate of the two rounds of expert consultation for valid questionnaires were all over 99％and the expert authority coefficients were all over 0.87.Conclusions The consulted experts have good enthusiasm and authority;the developed framework comprehensively covers training content,laying a foundation for designing and implementing tiered training programs for operating room nurses.

Objective To identify S-palmitoylation sites on angiotensin-converting enzyme 2(ACE2),the cellular receptor for severe acute respiratory syndrome corona virus(SARS-CoV)and SARS-CoV-2,and to investigate the functional relevance of these modifications in regulating ACE2 localization and activity.Methods An optimized multi-site mutagenesis strategy was performed by simultaneously substitution of all eight cysteine(Cys)residuesin ACE2 by serine to create a non-palmitoylatable mutant(8CS).Then individual cysteine residues were re-introduced one by one.Palmitoylation level of mutants was evaluated using a bioorthogonal click chemistry method to identify palmitoylation-competent residues.Immune-fluorescence staining and extra-cellular vesicle isolation assays were then used to evaluate the impact of specific palmitoylation sites on ACE2 sub-cellular localization.Results An efficient strategy for multi-site palmitoylation site mapping was optimized and successfully identified three critical palmitoylation sites on ACE2:Cys141,Cys344 and Cys498.Functional analyses showed that palmi-toylation of these sites significantly promotes the enrichment of ACE2 in extra-cellular vesicles.Conclusions S-palmitoylation at Cys141,Cys344 and Cys498 is essential for the trafficking of ACE2 to extra-cellular vesicles,which suggests a potential regulatory mechanism of its impact on viral receptor presentation and ACE2-associated signaling pathways.

Objective To investigate the clinical efficacy of fasting nasogastric administration of sesame oil combined with multi-frequency vibrational abdominal massage in stroke patients with consti-pation undergoing nasogastric feeding.Methods A total of 50 stroke patients with nasogastric feeding were selected as study subjects and randomly divided into control group and study group,with 25 pa-tients in each group.The control group received routine nursing care plus multi-frequency vibrational abdominal massage,while the study group received fasting nasogastric administration of 20 mL of sesa-me oil on the basis of the control group's treatment.After 15 days of intervention,the defecation condi-tions[constipation clinical symptom score(CSS),stool consistency(Bristol stool scale),and stool volume]were compared between the two groups.Results After treatment,the CSS score in the study group was lower than that in the control group[(6.52±2.52)versus(12.64±3.32),P＜0.05].The normal stool consistency(types Ⅳ and Ⅴ on the Bristol stool scale)in the study group was higher than that in the control group(76.0％versus 8.0％,P＜0.05).The stool volume in the study group was higher than that in the control group[(303.00±93.79)g versus(196.40±60.27)g,P＜0.05].The total effective rate of defecation in the study group was 96.0％,which was higher than 32.0％in the control group(P＜0.05).The Patient Assessment of Constipation-Quality of Life(PAC-QOL)score in the study group was lower than that in the control group[(40.07±5.67)versus(63.07±7.46),P＜0.05].Conclusion Fasting nasogastric administration of sesame oil com-bined with multi-frequency vibrational abdominal massage can promote the recovery of intestinal function and improve constipation symptoms in stroke patients with nasogastric feeding.