Background Long working hours, as a common risk factor for occupational stress, is closely related to the occurrence of depressive symptoms. Understanding how long working hours affect occupational stress and depressive symptoms will inform occupational health interventions. Objective To quantify the impact of long working hours exposure on occupational stress and depressive symptoms among Internet industry employees, translate black-box outputs into actionable insights, and demonstrate the value of interpretable machine learning for early-warning occupational-health surveillance. Methods A dataset was derived from a cross-sectional survey involving 2866 internet industry employees in China. This survey was part of the project Risk Assessment Of Long Working Hour Exposure And Its Adverse Health Effects, conducted by the National Institute for Occupational Health and Poisoning Control, Chinese Center for Disease Control and Prevention, from 2021 to 2023. Working hours, occupational stress and depressive symptoms were quantified with a set of structured questionnaires including the Core Occupational Stress Scale and the Patient Health Questionnaire. Pairwise associations were screened by Mantel tests and variance-inflation factors. Key predictors identified through feature selection were fed into six machine-learning risk-prediction models. Visual interpretation was provided by feature importance, Shapley additive explanations (SHAP) and local interpretable model-agnostic explanations (LIME), while directed causal effects and intervention impacts of prolonged working hours exposure on occupational stress and depressive symptoms were dissected with causal explanation of features techniques. Results The positive rates of occupational stress and depressive symptoms among internet employees were 12.9% and 77.8% respectively. Twelve core features for occupational stress and nine for depressive symptoms were retained after selection. After these features were supplied to six predictive algorithms and evaluated on five metrics, the Light Gradient Boosting Machine (LGBM) achieved the highest accuracy—0.89 for occupational stress and 0.79 for depressive symptoms on the hold-out test set. The feature-importance rankings converged on fatigue accumulation and life satisfaction as dominant drivers for both outcomes, whereas weekly working hours and daily overtime emerged as the principal exposure-related predictors. The SHAP summary plots revealed that longer weekly hours and daily overtime systematically elevated the probability of occupational stress. The causal feature explanation further quantified that ascending one category in weekly working hours increased the probability of occupational stress by 7.04%. Conclusion Exposure to long working hours is associated with both occupational stress and depressive symptoms among internet industry employees. Interpretable machine-learning frameworks translate these associations into transparent, defensible drivers, enabling precise identification of the pivotal factors and their interplay. This evidence base equips occupational-health practitioners with actionable insights for designing targeted prevention and intervention strategies.

A mounting body of research suggests that circRNAs significantly contribute to the develop-ment of myocardial fibrosis.The microarray results of human circular RNA expression profile indicated that circHERC4_041 expression increased in the myocardium of patients with heart failure,RT-qPCR a-nalysis confirmed that the myocardial expression level of circHERC4_041 in individuals with heart failure were considerably elevated compared to that in healthy organ donors.Fluorescence in situ hybridization(FISH)confirmed that circHERC4_041 was abundant in the cytoplasm of human cardiomyocyte AC16.Overexpression of circHERC4_041 in mouse myocardial fibroblasts(mCFs)mediated by adenovirus in-hibited the expression of fibrosis-related proteins in mCFs.Experiments involving cell proliferation,wound healing,and Transwell assays demonstrated that overexpression of circHERC4_041 suppressed the growth and mobility of mCFs(P＜0.001).Sequence analysis results suggested that circHERC4_041 con-tains potential ribosome entry sequence(IRES)and open reading frame(ORF).Western blot confirmed that circHERC4_041 could translate the 516 amino acid HERC4-516aa protein,which was mainly located in the cytoplasm of the cell.Cell functional experiments confirmed that circHERC4_041 inhibited the fi-brotic phenotype of mCFs by specifically translating HERC4-516aa(P＜0.05).The specific interaction between HERC4-516aa and transglutaminase 2(TGM2)was confirmed by IP-MS screening and Co-IP i-dentification.Further results found that the degradation of TGM2 was promoted through proteasome path-way.The overexpression of TGM2 in mCFs facilitated by adenoviral vectors could counteract the suppres-sive effects of HERC4-516aa on the fibrotic phenotype of mCFs.Therefore,this study confirmed that the HERC4-516aa protein translated by circHERC4_041 can specifically bind to TGM2 to inhibit the fibrotic phenotype of myocardial fibroblasts.

A mounting body of research suggests that circRNAs significantly contribute to the develop-ment of myocardial fibrosis.The microarray results of human circular RNA expression profile indicated that circHERC4_041 expression increased in the myocardium of patients with heart failure,RT-qPCR a-nalysis confirmed that the myocardial expression level of circHERC4_041 in individuals with heart failure were considerably elevated compared to that in healthy organ donors.Fluorescence in situ hybridization(FISH)confirmed that circHERC4_041 was abundant in the cytoplasm of human cardiomyocyte AC16.Overexpression of circHERC4_041 in mouse myocardial fibroblasts(mCFs)mediated by adenovirus in-hibited the expression of fibrosis-related proteins in mCFs.Experiments involving cell proliferation,wound healing,and Transwell assays demonstrated that overexpression of circHERC4_041 suppressed the growth and mobility of mCFs(P＜0.001).Sequence analysis results suggested that circHERC4_041 con-tains potential ribosome entry sequence(IRES)and open reading frame(ORF).Western blot confirmed that circHERC4_041 could translate the 516 amino acid HERC4-516aa protein,which was mainly located in the cytoplasm of the cell.Cell functional experiments confirmed that circHERC4_041 inhibited the fi-brotic phenotype of mCFs by specifically translating HERC4-516aa(P＜0.05).The specific interaction between HERC4-516aa and transglutaminase 2(TGM2)was confirmed by IP-MS screening and Co-IP i-dentification.Further results found that the degradation of TGM2 was promoted through proteasome path-way.The overexpression of TGM2 in mCFs facilitated by adenoviral vectors could counteract the suppres-sive effects of HERC4-516aa on the fibrotic phenotype of mCFs.Therefore,this study confirmed that the HERC4-516aa protein translated by circHERC4_041 can specifically bind to TGM2 to inhibit the fibrotic phenotype of myocardial fibroblasts.

Aim To construct and analyze the genotype of G protein-coupled receptor kinase 2(GRK2)conditional knockout mice in synoviocytes,and to provide an animal model for stud-ying the function of GRK2 in synoviocytes.Methods Grk2flox/+mice were bred to generate Grk2flox/flox mice,Grk2flox/flox mice were bred to Col1a1-iCre+mice,Grk2flox/+Col1a1-iCre+mice were bred to Grk2flox/flox mice.Grk2flox/flox Col1a1-iCre+mice were ob-tained as target mice.DNA was extracted and amplified by PCR to identify the genotype.Western blot was used to verify the effect of Grk2 knockout in synovium,liver and kidney tissues.HE staining was used to detect the effects of Grk2 conditional knockout in synovial cells on ankle synovium,liver and kidney tissues.Multiple immunofluorescence was used to detect GRK2 expression in synovial cells.Results The results of gene iden-tification showed that Grk2flox/flox Col1a1-iCre+mice had both Flox and Col1a1-iCre genotypes.Western blot results showed that GRK2 expression decreased in synovial tissues of Grk2flox/flox Col1a1-iCre+mice,but there was no significant change in the expression of GRK2 in liver and kidney tissues.HE staining showed that Grk2flox/flox Col1a1-iCre+mice had no significant pathological changes in the ankle synovium,liver and kidney.The results of multiple immunofluorescence showed that GRK2 expression in synovial cells of Grk2flox/flox Col1a1-iCre+mice de-creased.Conclusion Grk2 conditional knockout mice in syno-viocytes are successfully constructed and identified,which pro-vides an animal model for further study of the role of GRK2 in synovial-related diseases.

Objective:To analyze the trend of colorectal cancer incidence and age changes in cancer registration areas of Jiangsu Province from 2009 to 2019.Methods:Based on the continuous and complete data of 16 cancer registries with qualified quality control in Jiangsu Province from 2009 to 2019, the crude incidence rate, age-standardized incidence rate by Segi World Standard Population (ASIRW), age-specific incidence rate, mean age at onset, standardized mean age at onset, standardized age-specific incidence proportion, and incidence proportion of the population over 60 years old of colorectal cancer were calculated. Joinpoint software was used to calculate the average annual percentage change (AAPC) of crude incidence rate, ASIRW, age-specific incidence rate, and incidence proportion of the population over 60 years, respectively. Birth cohort models were constructed to analyze the incidence of colorectal cancer and its trends in the population born from 1929 to 2019. Linear regression models were used to analyze the correlation between mean age at onset, standardized mean age at onset and the year of onset.Results:From 2009 to 2019, a total of 48 036 new cases of colorectal cancer were collected from 16 cancer registries in Jiangsu Province, including 27 508 males and 20 528 females. The crude incidence rate and ASIRW of colorectal cancer in Jiangsu Province increased from 19.00/100 000 and 12.32/100 000 in 2009 to 33.49/100 000 and 16.75/100 000 in 2019, respectively, showing a significant upward trend (CR: AAPC=5.99%, ASIRW: AAPC=3.54%, P<0.001). The increase of ASIRW was greater in males than that observed in females (males: AAPC=4.31%, females: AAPC=2.34%, P<0.001), and greater in rural areas than in urban areas (rural areas: AAPC=4.03%, urban areas: AAPC=3.13%, P<0.001). The incidence of people over 50 years old increased significantly by year, with the 60~69 age group exhibiting a more rapid increase ( AAPC=4.97%, P<0.05). In the birth cohort, the incidence increased rapidly in the population over 50 years with the passage of birth year, with AAPCs ranging from 1.76% to 7.05% ( P<0.05). From 2009 to 2019, the standardized mean age at onset of colorectal cancer increased by 0.10 years annually. The proportion of standardized age-specific incidence exhibited a trend of increase in older age groups, and the incidence proportion of the population over 60 years old showed a significant yearly increase ( AAPC=0.86%, P<0.001). Conclusion:From 2009 to 2019, the incidence, mean age at onset and the incidence proportion of the population over 60 years old of colorectal cancer in Jiangsu Province could exhibit a rapid upward trend. The increase is particularly pronounced in males and rural areas. Additionally, the age-specific incidence distribution reveals a trend of increase in older age groups. Therefore, targeted adjustments and comprehensive prevention measures should be strengthened.

BACKGROUND:Alpinetin has anti-inflammatory,antitumor and antibacterial effects and has been proven to alleviate osteoporosis,but the effect and mechanism of alpinetin on osteoporotic fractures are still unclear.OBJECTIVE:To investigate the improvement effect of alpinetin in rates with osteoporotic fracture by regulating the cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA)/cAMP-responsive element binding protein (CREB) signaling pathway.METHODS:A rat model of osteoporotic fracture was constructed using bilateral ovariectomy. The successfully modeled rats were randomly separated into model group,low-,medium-,and high-dose alpinetin group,and inhibitor group,with 12 rats per group. Another 12 rats were regarded as the sham operation group. On the day of fracture,rats in the low-,medium-,and high-dose alpinetin groups were gavaged with 7.5,15,and 30 mg/kg alpinetin+intraperitoneally injected with an equal amount of normal saline,respectively;the inhibitor group was gavaged with 30 mg/kg alpinetin+intraperitoneally injected with 5 mg/kg H-89 (a pathway inhibitor);and the model and sham operation groups were given (gavaged+intraperitoneally injected) an equal amount of normal saline,once a day for 8 consecutive weeks. Radiological examination was applied to evaluate fracture healing and assess healing scores;bone mineral density scanners were used to measure bone mineral density at the fracture site;three-point bending experiment and compression experiment were conducted to evaluate the biomechanical status of the femur;pathological damage at the fracture site in rats was observed by hematoxylin-eosin staining;enzyme linked immunosorbent assay (ELISA) was used to detect changes in serum alkaline phosphatase,osteocalcin,type Ⅰ collagen cross-linked C-terminal peptide,and cAMP levels;western blot was applied to detect the expression of bone morphogenetic protein 2 and cAMP/PKA/CREB pathway proteins in femoral tissue.RESULTS AND CONCLUSION:Compared with the sham operation group,the fracture healing score,bone mineral density,maximum load,maximum stress,alkaline phosphatase,osteocalcin,cAMP levels,bone morphogenetic protein 2,p-PKA/PKA,p-CREB/CREB expression in the model group were decreased,the level of type Ⅰ collagen cross-linked C-terminal peptide was increased (P<0.05). The above indicators showed opposite changes in all the alpinetin groups compared with the model group (P<0.05). Compared with the high-dose alpinetin group,the changes in the above indicators were reversed in the inhibitor group (P<0.05). To conclude,alpinetin may accelerate fracture healing in osteoporotic fracture rats,by activating the cAMP/PKA/CREB signaling pathway.

Objective:To investigate the high-risk factors for early postoperative recurrence in patients with colorectal cancer liver metastasis (CRLM).Methods:The clinical data of 156 CRLM patients who underwent hepatic metastasectomy at the Li Huili Hospital, Ningbo University from Jan 2015 to Dec 2021 was retrospectively analyzed.Results:Among 156 patients, the postoperative recurrence rate was 55.8% (87/156).There were significant differences ( P<0.05) in the primary tumor site, T stage, TBS score, preoperative concentrations of CEA, CA199 and CA125, the maximum diameter of metastatic tumors and the number of metastatic tumors between the group with recurrence in one year and the group without recurrence in one year. Multivariate analysis showed that preoperative CA125 concentration ( OR=1.021, P=0.020), CEA concentration ( OR=1.044, P=0.018), and tumor burden score ( OR=3.067, P=0.011) were high risk factors influencing early recurrence ( P<0.05). In the low TBS score group(≤4), among the 49 patients who underwent simultaneous resection, 13 suffered from recurrence within 1 year, with a recurrence rate of 26.5%. Meanwhile, among the 29 patients who underwent staged resection, 17 suffered from recurrence within 1 year, with a recurrence rate of 58.6% ( P<0.05). In the high TBS score group(>4), among the 51 patients who underwent simultaneous resection, 37 suffered from recurrence within 1 year, with a recurrence rate of 72.5%, while in the 27 patients who underwent staged resection, 20 suffered from recurrence within 1 year, with a recurrence rate of 74.1% ( P>0.05). Conclusions:Preoperative TBS score, preoperative CA125 concentration, and CEA concentration are high-risk factors for postoperative recurrence in patients with colorectal liver metastases, simultaneous resection in patients with a low TBS score can reduce the risk of recurrence within 1 year after surgery.

OBJECTIVE To investigate the effect and potential mechanism of Ziyin Mingmu Formula in treating age-related mac-ular degeneration(AMD)by combining network pharmacology with animal model validation.METHODS Active ingredients of Ziyin Mingmu Formula were obtained from the TCMSP and BATMAN databases,and their targets were searched using the PharmMapper da-tabase.AMD disease targets were identified using the GeneCards,DrugBank,OMIM,and TTD databases.Venny analysis was per-formed to identify the intersection of active ingredient and disease targets.A protein-protein interaction(PPI)network was constructed using the String database,and core targets were screened using Cytoscape 3.9.0 software.KEGG pathway enrichment analysis was per-formed using the DAVID database.Molecular docking of key active ingredients with core targets was performed using Autodock software.A laser-induced mouse choroidal neovascularization(CNV)model was used.Optical coherence tomography angiography(OCTA)was used to assess CNV area in vivo,immunofluorescence staining was used to assess CNV area on choroidal flat mounts,and Western blot analysis was used to examine the expression of proteins involved in the vascular endothelial growth factor(VEGF)pathway.RESULTS Network pharmacology analysis identified 221 active ingredients in the Ziyin Mingmu Formula.PPI analysis i-dentified 29 core targets,including SRC,protein kinase B(AKT1),mitogen-activated protein kinase 1(MAPK1),and heat shock protein 90α family class A member 1(HSP90AA1).KEGG analysis revealed that the VEGF signaling pathway was the most highly en-riched.Molecular docking revealed that the core targets SRC,AKT1,MAPK1,and HSP90AA1 had good binding affinity with the main active ingredients,diosmetin,catechin,vestitol,and licochalcone A.Animal experiments showed that the Ziyin Mingmu Formula significantly reduced CNV area in model mice,downregulated VEGF protein expression,decreased VEGFR2,p38,and ERK1/2 pro-tein phosphorylation levels,and inhibited the VEGF signaling pathway.CONCLUSION Ziyin Mingmu Formula may inhibit CNV for-mation by regulating the VEGF signaling pathway.

BACKGROUND:Alpinetin has anti-inflammatory,antitumor and antibacterial effects and has been proven to alleviate osteoporosis,but the effect and mechanism of alpinetin on osteoporotic fractures are still unclear.OBJECTIVE:To investigate the improvement effect of alpinetin in rates with osteoporotic fracture by regulating the cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA)/cAMP-responsive element binding protein (CREB) signaling pathway.METHODS:A rat model of osteoporotic fracture was constructed using bilateral ovariectomy. The successfully modeled rats were randomly separated into model group,low-,medium-,and high-dose alpinetin group,and inhibitor group,with 12 rats per group. Another 12 rats were regarded as the sham operation group. On the day of fracture,rats in the low-,medium-,and high-dose alpinetin groups were gavaged with 7.5,15,and 30 mg/kg alpinetin+intraperitoneally injected with an equal amount of normal saline,respectively;the inhibitor group was gavaged with 30 mg/kg alpinetin+intraperitoneally injected with 5 mg/kg H-89 (a pathway inhibitor);and the model and sham operation groups were given (gavaged+intraperitoneally injected) an equal amount of normal saline,once a day for 8 consecutive weeks. Radiological examination was applied to evaluate fracture healing and assess healing scores;bone mineral density scanners were used to measure bone mineral density at the fracture site;three-point bending experiment and compression experiment were conducted to evaluate the biomechanical status of the femur;pathological damage at the fracture site in rats was observed by hematoxylin-eosin staining;enzyme linked immunosorbent assay (ELISA) was used to detect changes in serum alkaline phosphatase,osteocalcin,type Ⅰ collagen cross-linked C-terminal peptide,and cAMP levels;western blot was applied to detect the expression of bone morphogenetic protein 2 and cAMP/PKA/CREB pathway proteins in femoral tissue.RESULTS AND CONCLUSION:Compared with the sham operation group,the fracture healing score,bone mineral density,maximum load,maximum stress,alkaline phosphatase,osteocalcin,cAMP levels,bone morphogenetic protein 2,p-PKA/PKA,p-CREB/CREB expression in the model group were decreased,the level of type Ⅰ collagen cross-linked C-terminal peptide was increased (P<0.05). The above indicators showed opposite changes in all the alpinetin groups compared with the model group (P<0.05). Compared with the high-dose alpinetin group,the changes in the above indicators were reversed in the inhibitor group (P<0.05). To conclude,alpinetin may accelerate fracture healing in osteoporotic fracture rats,by activating the cAMP/PKA/CREB signaling pathway.

Objective:To investigate the electron microscopic evaluation results of pre-transplant biopsies of deceased donor kidneys and the corresponding recovery of graft function in recipients after kidney transplantation.Methods:A retrospective analysis was conducted on the clinical data and donor kidney electron microscopy (EM) reports of 196 kidney transplant recipients who underwent pre-implantation kidney biopsies at Renmin Hospital of Wuhan University between October 2020 and June 2023. The ultrastructural pathological features assessed in the pre-transplant kidney biopsy included: the number of glomeruli, the presence of leukocytes and endothelial cells within the glomeruli, the arrangement of capillary loops, abnormalities in podocytes, mesangial cells, mesangial matrix and glomerular basement membrane (GBM), and multilayering of the peritubular capillary basement membrane. Referring to the Mayo Clinic/Renal Pathology Society Consensus Report on the pathological classification, diagnosis, and reporting of glomerulonephritis, the expert consensus on renal biopsy pathology reporting models, and the content of the 2019 Banff Transplant Pathology Meeting, a scoring system was established for ultrastructural pathology of donor kidneys. According to the scores, the recipients were divided into three groups: Group A (total score ≥ 8 points, 94 cases), Group B (total score between 6 and 8 points, 85 cases), and Group C (total score < 6 points, 17 cases). The serum creatinine, estimated glomerular filtration rate (eGFR), and proteinuria at postoperative day 1, day 7, month 1, month 3, month 6, and year 1 were compared among the three groups.Results:Among the 196 donor kidneys, EM examination before transplantation showed the following findings: 19 cases (9.7%) had prominent leukocyte infiltration within the glomeruli, 8 cases (4.1%) had mild leukocyte infiltration, and the remaining 169 cases (86.2%) showed no obvious increase in glomerular leukocytes. Glomerular capillary loop collapse or narrowing was seen in 19 cases (9.7%). Endothelial cell proliferation was observed in 32 cases (16.3%). Homogeneous thickening of the GBM (400-900 nm) was present in 82 cases (41.8%). Foot process effacement ranged from partial to diffuse. Podocyte vacuolar degeneration was seen in 62 cases (32.1%), and podocyte swelling in 10 cases (5.1%). Electron-dense deposits in the mesangial area were observed in 9 cases (4.6%). Mild tubular epithelial swelling was present in 6 cases (3.1%). Only 22 cases (11.2%) showed no multilayering of the peritubular capillary basement membrane, while 174 cases (88.8%) showed 2 to 5 layers of multilayering. All donor glomeruli showed irregular capillary loop arrangement, and the renal interstitium showed scattered inflammatory infiltration and varying degrees of collagen fiber deposition. Group C had significantly higher qualitative proteinuria levels at day 7 and month 1 post-transplantation compared with Groups A and B ( P=0.036, 0.004). There were no statistically significant differences in other renal function indicators among the groups (all P>0.05). Conclusions:Pre-transplant electron microscopy of donor kidneys helps in accurately assessing donor kidney quality and identifying subtle pre-existing pathological changes. It can serve as a reference tool for predicting post-transplant functional recovery. Mild ultrastructural abnormalities in donor kidneys have a relatively controllable impact on long-term graft outcomes.