Objective To identify long non-coding RNA (lncRNA) associated with rheumatoid arthritis-associated interstitial lung disease (RA-ILD) and investigate their mechanisms. Methods Peripheral blood samples were collected from RA-ILD patients (n＝3), RA patients without lung involvement (n＝3), and healthy controls (n＝3). Next-generation sequencing was performed to screen differentially expressed lncRNA. A human fibrotic lung cell model was established by inducing the MRC-5 cell line with transforming growth factor-β (TGF-β). Following siRNA-mediated knockdown of target genes, changes in inflammatory and oxidative stress-related genes were analyzed via real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Western blotting and dual-luciferase reporter (DLR) assays were used to validate protein expression, ubiquitination levels, and nuclear translocation of oxidative stress regulators, and antioxidant response element (ARE) transcriptional activity. Rescue experiments were conducted to confirm the role of target lncRNA in oxidative stress and inflammation in fibrotic lung cells. Results High-throughput sequencing revealed significant downregulation of LINC00638 in RA-ILD patients. Knockdown of LINC00638 markedly reduced transcriptional levels of interleukin (IL)-4, nuclear factor erythroid-2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and superoxide dismutase 2 (SOD2), while increasing IL-6, IL-1β, interferon-γ (IFN-γ), and reactive oxygen species (ROS) levels. Furthermore, LINC00638 knockdown decreased Nrf2 protein expression, increased its ubiquitination, reduced nuclear translocation, and suppressed ARE transcriptional activity. In MRC-5 cells, LINC00638 knockdown combined with N-acetylcysteine treatment restored Nrf2 and HO-1 levels while reducing IL-6 expression. Conclusions LINC00638 suppresses inflammatory responses in RA-ILD by activating the Nrf2/ARE antioxidant signaling pathway, suggesting its potential as a therapeutic target for diagnosis and treatment.

Objective To investigate the molecular mechanism of hepatic fibrosis(HF)regulation by liver sinusoidal endothelial cells(LSECs)via endothelial mesenchymal transition(EnMT),and to predict the natural active components using bioinformatics,machine learning,and cellular experiments.Methods HF and EnMT gene matrices were obtained and the intersecting genes were extracted and enriched using Limma difference analysis and weighted gene co-expression network analysis(WGCNA).The diagnostic genes were screened using a combination of random forest method,support vector machine-recursive feature elimination and network topology analysis,and immune infiltration analysis and prediction of natural active ingredients were performed.The expression of diagnostic genes and the pharmacological effects of the predicted ingredients were finally verified by cellular experiments.Results Differential analysis yielded 3034 EnMT-associated and 4133 HF-associated differential genes.WGCNA analysis yielded 4589 EnMT-associated Hub genes and 763 HF-associated Hub genes.Thirty-eight intersecting genes were extracted,which were mainly enriched in the pathways of basement membrane and extracellular matrix receptor interaction.Four diagnostic genes,CFP,COL4A2,ITGA1,and GRPEL1,were screened by multidimensional analysis.Immune infiltration analysis showed that the diagnostic genes were closely associated with mast cell resting state,memory B cells,and memory CD4+T cells.Reverse transcription-polymerase chain reaction analysis showed significantly increased mRNA expression levels of the four diagnostic genes in the Jagged1-induced model group(P＜0.05).The predicted components,sterol,kaempferol,and quercetin,all had good binding activities with the diagnostic genes.Enzyme-linked immunosorbent assay result confirmed that all three active components significantly reduced the expression of collagen type Ⅳ α2 chain protein in Jagged1-induced LSECs,with quercetin having the most significant effect(P＜0.01).Conclusions This study elucidated the molecular mechanism of hepatic sinusoidal endothelial cells involved in the pathological process of HF through mesenchymal transition.We also propose a diagnostic marker system including CFP,COL4A2,ITGA1,and GRPEL1 as core genes.The result also suggest that natural active ingredients,such as quercetin,may exert anti-HF pharmacological effects by targeting these diagnostic genes.

Objective To systematically compare the perioperative outcomes of robot-assisted transvaginal natural orifice transluminal endoscopic surgery total hysterectomy(R-vNOTES-TH),transvaginal natural orifice transluminal endoscopic surgery total hysterectomy(vNOTES-TH),and robot-assisted laparoendoscopic single-site total hysterectomy(R-LESS-TH),and to evaluate the clinical advantages of R-vNOTES-TH.Methods Clinical data of 259 patients undergoing total hysterectomy for benign diseases at Zhongnan Hospital of Wuhan University from Jan.2020 to Dec.2024 were retrospectively analyzed.Baseline indicators and perioperative indicators were collected.Patients were assigned to 3 groups according to the surgical approach:R-vNOTES-TH group(n=22),vNOTES-TH group(n=39),or R-LESS-TH group(n=198).Perioperative indicators were compared between the R-vNOTES-TH group and the other 2 groups to evaluate the advantages of R-vNOTES-TH.Results Compared with the vNOTES-TH group,the R-vNOTES-TH group had significantly less intraoperative blood loss(50[50,100]mL vs 100[50,100]mL,P=0.027),lower intraoperative fluid infusion volume(1 000[500,1 000]mL vs 1 000[1 000,1 500]mL,P＜0.001),and shorter urinary catheter indwelling time(3[1,4]d vs 5[2,5]d,P=0.043),but longer vaginal drain indwelling time(2[2,3]d vs 2[0,2]d,P=0.004).Compared with the R-LESS-TH group,the R-vNOTES-TH group had longer urinary catheter indwelling time(3[1,4]d vs 1[1,1]d,P＜0.001).Conclusion Compared with vNOTES-TH,R-vNOTES-TH enhances intraoperative operational precision,reduces bleed loss,and accelerates urinary catheter removal,confirming that the robotic system effectively overcomes the technical limitations of conventional vNOTES.Although R-vNOTES-TH eliminates abdominal wall trauma-thereby prolonging urinary catheter indwelling time relative to R-LESS-TH-it offers patients a truly scar-free alternative.

AIM:To investigate the protective mechanism of calycosin(CA)on renal structure and antioxi-dant capacity in acute kidney injury(AKI)mice.METHODS:Thirty male C57BL/6J mice were allocated into control group,model group,and treatment group.The treatment group was further subdivided into low-dose CA,middle-dose CA,and high-dose CA groups,receiving 5 mg/kg,10 mg/kg,and 20 mg/kg CA solution by gavage for 6 days.After the pre-treatment phase,both the model and treatment groups were administered an intraperitoneal injection of 20 mg/kg of cis-platin(CDDP)to induce AKI modeling.Specimens from the mice were collected 72 hours post-modeling for further analy-sis.The body weight and the mean mass of both kidneys were assessed.Serum creatinine(SCr)and blood urea nitrogen(BUN)levels were assessed to evaluate kidney function.Kidney pathology was assessed with periodic acid-Schiff(PAS)staining,while protein levels of superoxide dismutase 1(SOD1),SOD2,glutathione peroxidase 1(GPX1),and catalase were evaluated using Western blot and immunohistochemistry assay.RESULTS:Compared with the control group,the model group exhibited a substantial reduction in body mass,a significant increase in the mean mass of bilateral kidneys,and elevated levels of SCr and BUN,indicating impaired renal function(P<0.01).PAS staining revealed pronounced structural damage to the glomeruli and renal tubules,with widespread appearance of tubular casts in model group.The re-sults of Western blot and immunohistochemical staining demonstrated significantly reduced expression of antioxidant en-zymes SOD1,SOD2,GPX1,and catalase(P<0.05).The body weight of mice in the low-dose and middle-dose CA groups exhibited a significant increase than those in the model group,and the kidney weights of mice in Low-dose,middle-dose,and high-dose CA groups were significantly lower than those of mice in the model group(P<0.05).Additionally,SCr and BUN levels were significantly reduced(P<0.01).PAS staining indicated obvious recovery of glomeruli and renal tubules after CA treatment.SOD1 and GPX1 levels were elevated in the middle-dose and high-dose CA groups,and SOD2 and catalase levels were significantly higher in the middle-dose CA group(P<0.05),compared with model group.CON-CLUSION:CA can alleviate CDDP-induced kidney injury in AKI mice by enhancing the expression of antioxidant enzymes SOD1,SOD2,GPX1,and catalase.

Objective To investigate the molecular mechanism of hepatic fibrosis(HF)regulation by liver sinusoidal endothelial cells(LSECs)via endothelial mesenchymal transition(EnMT),and to predict the natural active components using bioinformatics,machine learning,and cellular experiments.Methods HF and EnMT gene matrices were obtained and the intersecting genes were extracted and enriched using Limma difference analysis and weighted gene co-expression network analysis(WGCNA).The diagnostic genes were screened using a combination of random forest method,support vector machine-recursive feature elimination and network topology analysis,and immune infiltration analysis and prediction of natural active ingredients were performed.The expression of diagnostic genes and the pharmacological effects of the predicted ingredients were finally verified by cellular experiments.Results Differential analysis yielded 3034 EnMT-associated and 4133 HF-associated differential genes.WGCNA analysis yielded 4589 EnMT-associated Hub genes and 763 HF-associated Hub genes.Thirty-eight intersecting genes were extracted,which were mainly enriched in the pathways of basement membrane and extracellular matrix receptor interaction.Four diagnostic genes,CFP,COL4A2,ITGA1,and GRPEL1,were screened by multidimensional analysis.Immune infiltration analysis showed that the diagnostic genes were closely associated with mast cell resting state,memory B cells,and memory CD4+T cells.Reverse transcription-polymerase chain reaction analysis showed significantly increased mRNA expression levels of the four diagnostic genes in the Jagged1-induced model group(P＜0.05).The predicted components,sterol,kaempferol,and quercetin,all had good binding activities with the diagnostic genes.Enzyme-linked immunosorbent assay result confirmed that all three active components significantly reduced the expression of collagen type Ⅳ α2 chain protein in Jagged1-induced LSECs,with quercetin having the most significant effect(P＜0.01).Conclusions This study elucidated the molecular mechanism of hepatic sinusoidal endothelial cells involved in the pathological process of HF through mesenchymal transition.We also propose a diagnostic marker system including CFP,COL4A2,ITGA1,and GRPEL1 as core genes.The result also suggest that natural active ingredients,such as quercetin,may exert anti-HF pharmacological effects by targeting these diagnostic genes.

AIM:To investigate the protective mechanism of calycosin(CA)on renal structure and antioxi-dant capacity in acute kidney injury(AKI)mice.METHODS:Thirty male C57BL/6J mice were allocated into control group,model group,and treatment group.The treatment group was further subdivided into low-dose CA,middle-dose CA,and high-dose CA groups,receiving 5 mg/kg,10 mg/kg,and 20 mg/kg CA solution by gavage for 6 days.After the pre-treatment phase,both the model and treatment groups were administered an intraperitoneal injection of 20 mg/kg of cis-platin(CDDP)to induce AKI modeling.Specimens from the mice were collected 72 hours post-modeling for further analy-sis.The body weight and the mean mass of both kidneys were assessed.Serum creatinine(SCr)and blood urea nitrogen(BUN)levels were assessed to evaluate kidney function.Kidney pathology was assessed with periodic acid-Schiff(PAS)staining,while protein levels of superoxide dismutase 1(SOD1),SOD2,glutathione peroxidase 1(GPX1),and catalase were evaluated using Western blot and immunohistochemistry assay.RESULTS:Compared with the control group,the model group exhibited a substantial reduction in body mass,a significant increase in the mean mass of bilateral kidneys,and elevated levels of SCr and BUN,indicating impaired renal function(P<0.01).PAS staining revealed pronounced structural damage to the glomeruli and renal tubules,with widespread appearance of tubular casts in model group.The re-sults of Western blot and immunohistochemical staining demonstrated significantly reduced expression of antioxidant en-zymes SOD1,SOD2,GPX1,and catalase(P<0.05).The body weight of mice in the low-dose and middle-dose CA groups exhibited a significant increase than those in the model group,and the kidney weights of mice in Low-dose,middle-dose,and high-dose CA groups were significantly lower than those of mice in the model group(P<0.05).Additionally,SCr and BUN levels were significantly reduced(P<0.01).PAS staining indicated obvious recovery of glomeruli and renal tubules after CA treatment.SOD1 and GPX1 levels were elevated in the middle-dose and high-dose CA groups,and SOD2 and catalase levels were significantly higher in the middle-dose CA group(P<0.05),compared with model group.CON-CLUSION:CA can alleviate CDDP-induced kidney injury in AKI mice by enhancing the expression of antioxidant enzymes SOD1,SOD2,GPX1,and catalase.

Objective To investigate the epidemiological characteristics and spatial distribution characteristics of Clonorchis sinensis human infections in Guangdong Province from 2016 to 2022, so as to provide insights into formulation of the clonorchiasis control measures in the province. Methods Xinhui District of Jiangmen City, Longmen County of Huizhou City and Wengyuan County of Shaoguan City in Guangdong Province were selected as fixed surveillance sites for human clonorchiasis from 2016 to 2022, and additional 10% to 15% counties (districts) endemic for clonorchiasis were sampled from Guangdong Province as mobile surveillance sites each year from 2016 to 2022. A village (community) was randomly selected from each surveillance site according to the geographical orientations of east, west, south, north and middle, and subjects were randomly sampled from each village (community). C. sinensis eggs were detected in subjects’ stool samples using the Kato-Katz technique, and the prevalence and intensity of C. sinensis infections were calculated. In addition, subjects’ gender, age, ethnicity, educational level and occupation were collected. The Guangdong Provincial 1:1 million electronic map in vector format was downloaded from the National Geomatics Center of China, and kernel density analysis and spatial autocorrelation analysis of C. sinensis human infections in Guangdong Province from 2016 to 2022 were performed using the software ArcGIS 10.7. Results A total of 153 188 residents were tested for C. sinensis infections in Guangdong Province from 2016 to 2022, including 75 596 men (49.35%) and 77 592 women (50.65%), and there were 5 369 residents infected with C. sinensis, with 3.50% overall prevalence of infections. The prevalence rates of severe, moderate and mild C. sinensis infections were 0.76%, 7.26% and 91.97% among C. sinensis-infected residents in Guangdong Province from 2016 to 2022, and there were age-, gender-, ethnicity-, occupation- and educational level-specific prevalence of C. sinensis human infections (χ² = 2 578.31, 637.33, 52.22, 2 893.28 and 1 139.33, all P values < 0.05). Global spatial autocorrelation analysis showed a cluster in the prevalence of C. sinensis human infections in Guangdong Province (Moran’s I = 0.63, Z = 27.31, P < 0.05). Kernel density analysis showed that the prevalence of C. sinensis human infections with a high kernel density in Guangdong Province was mainly distributed along the Zhujiang River basin in Pearl River Delta areas, followed by in eastern and northern Guangdong Province. In addition, local spatial autocorrelation analysis identified 73 high-high clusters of the prevalence of C. sinensis human infections in Guangdong Province. Conclusions The prevalence of C. sinensis human infections was high in Guangdong Province from 2016 to 2022, and mild infection was predominant among all clonorchiasis cases, with spatial clusters identified in the prevalence of C. sinensis human infections. Targeted clonorchiasis control measures are required among high-risk populations and areas.

The theory of "liver opens at the eyes" was first seen in Yellow Emperor's Internal Canon of Medicine， which is the ancient people's summary of the connection between the liver and the eyes. The theory of "liver opens at the eyes" suggests the characteristic of "co-damage and co-recover of liver and eyes". It has been found in clinical practice that liver diseases and eye diseases often occur together， and "liver and eyes co-recover" is an ideal choice. The key to achieving "liver and eyes co-recover" is to analyze its pharmacological material basis and mechanism. With the development of modern medicine， more and more evidence indicates that the liver and eyes have complex and close relationships in physiological and pathological aspects. In a pathological state， there is a phenomenon of "liver and eyes co-damage"， and after the intervention of traditional Chinese medicine， "liver and eyes co-recover" occurs. "Liver and eyes co-damage and co-recover" can be explained through the "co-material basis and co-action mechanism". On this basis， the research group tentatively proposed that the liver and eyes had "four common characteristics" （4CCs）， namely "co-damage， co-recover， co-material basis， and co-action mechanism" from the theoretical connotation of traditional Chinese medicine， clinical practice， and molecular biology. Additionally， the group also took the intervention of Prunella vulgaris， traditional Chinese medicine， for removing liver fire and improving eyesight on immune liver injury （ILI） and allergic conjunctivitis （AC） as examples to analyze 4CCs. This project aims to deeply analyze the scientific connotation of the theory of "liver opens at the eyes"， reveal the common characteristics and biological essence of liver and eyes， explore a new research paradigm of "liver and eyes co-recover"， and provide a reference for the study of common problems of multi-organ associated diseases.

Objective:To investigate the knowledge, attitude and practice of general practitioners (GPs) in Shenzhen towards managing patients with common mental health problems.Methods:It was a cross-sectional study, from September 19 to October 31, 2020, 500 GPs from 100 community health centers (CHC) in 10 districts of Shenzhen municipality were randomly selected as the research subjects by stratified random sampling; the survey was conducted by self-filled questionnaire, which included general conditions, knowledge tests of common psychological problems (generalized anxiety disorder, depressive disorder, panic attacks, sleep disorders), GPs′ attitudes to take care of common psychological problems, and GPs′ practice of caring for common psychological problems.Results:A total of 500 questionnaires were distributed and 329 valid ones were collected. In terms of knowledge, GPs had the highest accuracy of answering the characteristics of depressive disorder (97.3%, 320/329), and the lowest accuracy of case analysis of panic attacks (50.2%, 165/329). In terms of attitude, 71.1%(234/329)of GPs agreed that "common psychological problems should be taken care of",there was a statistically significant difference in attitude scores among GPs with different years of working in CHC ( Z=14.60, P=0.006). In terms of practice, the most encountered mental health problem was insomnia (91.2%, 300/329), 46.5% (153/329) of GPs would use psychological assessment scales, 52.9% (174/329) of GPs would choose direct referral, and there were statistically significant differences in attitude scores among GPs with different years of working ( Z=10.70, P=0.030) and years of working in CHC ( Z=22.14, P<0.001). Conclusions:GPs have a positive attitude in taking care of common psychological problems, but lack of knowledge and confidence. As working in CHC for more years, GPs are more inclined to care for patients with common psychological problems in practice.

Objective:To investigate the clinical, skeletal muscle pathological, and genetic characteristics of fatal infantile hypertonic myofibrillar myopathy (FIHMM).Methods:The clinical manifestations, laboratory assessments data and gene sequencing results of 10 patients diagnosed with FIHMM in Shenzhen Children′s Hospital from February 2017 to April 2021 were retrospectively analyzed.Magnetic resonance imaging (MRI) of both musculoskeletal system and the brain, and electromyogram (EMG) were performed in 3 cases, while muscle biopsy was performed in 2 cases.Results:Among these 10 cases, 1 case was from Northeast China and 1 case from East China, while the rest 8 cases were from South China.Eight of the 10 patients were male, and the other 2 cases were female.They were all born normal and not related to each other.The age of onset varied from 2 to 12 months.The main clinical manifestations for all the patients were progressive rigidity of the rectus abdominis (8 cases), neck muscles (7 cases), rectus abdominis (2 cases) and intercostal muscles (1 case), resulting in respiratory failure.Mildly to moderately elevated serum creatine kinase level was detected (436-5 804 IU/L) (reference range: 24-229 IU/L). Complex repetitive discharges can be seen in the EMG, without any myotonic potential.Muscle fiber degeneration, necrosis, and vacuolar degeneration were noted in the histopathological examination of the vastus lateralis and rectus abdominis.An abnormal red granular deposit was observed in a portion of the field of the modified Gomory Trichrome staining.Immunohistochemistry showed substantial deposition of desmin.Under the electron microscopy, the sarcomere structure of the muscle fibers was seriously disordered, with the destruction of Z-bands and the presence of granular deposits.The whole-exome sequencing identified the same homozygous variation c. 3G>A, p.Met1? of CRYAB gene in all the patients, but heterozygous variation in their parents. Conclusions:Axial muscles involvement, such as rectus abdominis rigidity, is the main clinical characteristic of FIHMM.c.3G>A, p.Met1? mutation in the CRYAB gene is a hotspot mutation in Chinese children.