Objective To investigate the role and clinical significance of Calpain activity and 145 kDa cleavage fragments of αⅡ-spec-trin breakdown products(SBDP145)in systemic lupus erythematosus(SLE).Methods 124 patients with confirmed SLE and de-tailed clinical data were collected as the SLE group,and 75 healthy individuals who underwent physical examination during the same period were selected as the control group.Their serum samples were collected,and serum SBDP145 levels and Calpain activity were de-tected by the enzyme-linked immunosorbent assay(ELISA)and substrate-based method,respectively.The Mann-Whitney U test,Spearman rank correlation,univariate and multivariate Logistic regression,and receiver operating characteristic(ROC)curve were used to evaluate the clinical value of Calpain activity and SBDP145 in the diagnosis and assessment of SLE.Results Compared with healthy controls,serum Calpain activity([17.000[8.5000,33.500]RFU vs 7.500[4.000,12.500]RFU)and SBDP145 levels(5.283[3.532,9.463]ng/mL vs 1.472[0.994,2.212]ng/mL)in SLE patients were significantly increased(Z=-9.229,P<0.001;Z=-6.881,P<0.001).Furthermore,the Calpain activity was significantly correlated with SLE disease activity index(SLEDAI-2K)score(r=0.349,P<0.001).Logistic regression analysis showed that the increased Calpain activity and SBDP145 levels were significantly associated with the occurrence of SLE(OR=1.164,95％CI:1.016-1.334,P=0.029;OR=3.822,95％CI:1.928-7.574,P<0.001).The ROC curve analysis showed that the area under the ROC curve(AUCROC)of serum Calpain activity in distin-guishing SLE and healthy controls was 0.762(95％CI:0.697-0.827).When the cut-off value was 13.75 RFU,its sensitivity and speci-ficity were 63.1％and 81.1％,respectively.The AUCROC of serum SBDP145 levels in distinguishing SLE and healthy controls was 0.891(95％CI:0.845-0.936).When the cut-off value was 2.377 ng/mL,its sensitivity and specificity were 88.7％and 80.0％,re-spectively.The AUCROC,sensitivity,and specificity of combining the two with traditional SLE clinical indicators,including complement C3,erythrocyte sedimentation rate,and anti-dsDNA antibodies,in the diagnosis of SLE reached 0.986(95％CI:0.972-1.000),93.3％,and 96.0％,respectively.The Spearman correlation analysis results showed that the organ damage index(SDI)score of SLE patients was positively correlated with serum Calpain activity and SBDP145 levels(r=0.342,P<0.001;r=0.250,P=0.005).Con-clusion The elevated Calpain activity and SBDP145 levels are closely related to the occurrence and development of SLE,suggesting that they may be served as potential biomarkers for the diagnosis and assessment of SLE patients.

Objective:To investigate the role and mechanism of leucine-rich α-2-glycoprotein 1 (LRG1) in the fibrosis of human pterygium fibroblasts (HPFs).Methods:A total of 30 nasal primary pterygium tissues from patients who underwent pterygium excision surgery and 30 nasal normal conjunctival tissues from patients who underwent strabismus correction surgery were collected from the First Affiliated Hospital of Harbin Medical University between January 2022 and March 2023, serving as the pterygium group and normal control group, respectively.LRG1 protein expression in both groups was detected by immunofluorescence staining.The mRNA and protein levels of LRG1 and transforming growth factor-β1 (TGF-β1) were evaluated by quantitative real-time PCR (qRT-PCR) and Western blot.Primary HPFs were cultured from excised pterygium tissues using tissue block adhesion method, and cell morphology was observed.Vmentin and cytokeratin were identified by immunofluorescence staining.HPFs were divided into recombinant human LRG1 (rhLRG1) group and blank control group treated with or without 10 μg/ml rhLRG1 for 24 hours, respectively, and cell migration was evaluated via scratch assay.Additionally, HPFs were divided into blank control group, LRG1 overexpression group and LRG1 knockdown group.HPFs in LRG1 overexpression group and LRG1 knockdown group were transfected with LRG1 overexpression plasmids and small interfering RNA for 24 hours, respectively.TGF-β1 mRNA level was evaluated by qRT-PCR and expression of TGF-β1, fibronectin (FN), type Ⅲ collagen (COL3), and α-smooth muscle actin (α-SMA) proteins were evaluated by Western blot.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of the First Affiliated Hospital of Harbin Medical University (No.2022IIT026).Written informed consent was obtained from each subject.Results:HPFs were successfully isolated, exhibiting spindle-shaped morphology with whorled arrangement, positive identification for vimentin, and negative immunofluorescence staining for cytokeratin.The migration rate of the rhLRG1 group was (83.01±2.56)%, significantly higher than (50.32±4.97)% of the blank control group ( t=9.59, P<0.001).Immunofluorescence staining results showed that compared with normal conjunctival tissue, LRG1 protein was significantly higher expressed in pterygium tissue and was widely distributed in fibrous connective tissue and epithelial layer.Both mRNA and protein levels of LRG1 and TGF-β1 were significantly higher in the pterygium group than in the normal control group (mRNA: t=10.18, 6.15, both P<0.05.protein: t=6.83, 8.79, both P<0.05).In the LRG1 overexpression group, mRNA level of TGF-β1, and protein levels of FN, COL3 and α-SMA were significantly increased compared with the blank control and LRG1 knockdown groups (all P<0.05). Conclusions:LRG1 promotes fibrosis and enhances the migration ability in HPFs, and its mechanism may be associated with the upregulation of the TGF-β1 signaling pathway.

Objective:To analyze the prevalence and influencing factors of arteriosclerosis in different examination items in health check-up population.Methods:It was a cross-sectional study. A total of 4 479 subjects who underwent fundus, carotid artery color ultrasound and arteriosclerosis detector examinations in the Health Management Center at Tianjin Medical University General Hospital from January to December 2019 were selected as the study objects. The data including age, gender, family history and biochemical indicators were collected. The detection outcomes of arteriosclerosis and the distribution in different age and gender subgroups were analyzed. Kappa consistency test was used to analyze the consistency of different examination items of arteriosclerosis examination. The multiple logistic regression model was used to analyze the influencing factors of different examination items of arteriosclerosis detection. Results:Among the 4 479 subjects included in the analysis, 2 450 were male and 2 029 were female, with a mean age of (44.48±8.12) years. Arteriosclerosis was detected in one examination item in 1 167 cases (26.05%), 1 042 cases (23.26%) in two examination items, and 617 cases (13.78%) in three examination items, respectively. Among patients aged ≤40 years, 149 cases (18.91%) were detected with arteriosclerosis. In the age group of 40<-60 years, arteriosclerosis was detected in 2 660 patients (72.40%). All patients >60 years old exhibited arteriosclerosis, with 16 cases (94.12%) showing ≥2 items of arteriosclerosis. The results of fundus and carotid artery color Doppler ultrasound, fundus and arteriosclerosis detector, carotid artery color Doppler ultrasound and arteriosclerosis detector all demonstrated moderate consistency ( Kappa=0.32, 0.34, 0.24; all P<0.01). Logistic regression analysis revealed that the age ( OR=2.07, 95% CI: 1.97-2.17), male ( OR=1.60, 95% CI: 1.14-2.23), body mass index ( OR=1.10, 95% CI: 1.05-1.15), and systolic blood pressure ( OR=1.02, 95% CI: 1.01-1.03) were positively correlated with fundus arteriosclerosis (all P<0.05). Similarly, age ( OR=1.13, 95% CI: 1.12-1.14), male ( OR=2.53, 95% CI: 2.07-3.09), systolic blood pressure ( OR=1.02, 95% CI: 1.01-1.03), and low-density lipoprotein cholesterol ( OR=1.61, 95% CI: 1.16-2.23) were positively correlated with cervical arteriosclerosis, while high-density lipoprotein cholesterol ( OR=0.48, 95% CI: 0.32-0.72) was negatively correlated with cervical arteriosclerosis (all P<0.05). Furthermore, age ( OR=1.09, 95% CI: 1.08-1.11), male ( OR=1.55, 95% CI: 1.25-1.92), systolic blood pressure ( OR=1.08, 95% CI: 1.07-1.09), diastolic blood pressure ( OR=1.02, 95% CI: 1.00-1.03), postprandial blood glucose ( OR=1.09, 95% CI: 1.03-1.15), homocysteine ( OR=1.02, 95% CI: 1.00-1.03), and fibrinogen ( OR=1.26, 95% CI: 1.04-1.52) were positively correlated with systemic arterial stiffness (all P<0.05). Conclusions:The detection rates of arteriosclerosis in different examination items among the health check-up population vary. Age, male gender and systolic blood pressure are positive correlation factors for the three detection items of arteriosclerosis. For the examinees with aggregated risk factors, the multi-site and combined screening mode has certain significance for the early diagnosis and screening of arteriosclerotic diseases.

Objective:To explore the association between cumulative blood pressure (BP) and the risk of cerebrovascular disease (CVD) in older adults.Methods:This retrospective cohort study consecutively selected 4 480 older adults who participated in the Chinese Longitudinal Healthy Longevity Survey from 2008 to 2009 as the study subjects. The cumulative BP was calculated using the area under the curve from measurements taken at baseline (2008-2009), the first follow-up (2011), and the second follow-up (2014). The subjects were grouped with the quartiles (Q1-Q4) of cumulative systolic blood pressure (SBP), diastolic blood pressure (DBP), and pulse pressure (PP). The Cox proportional hazards regression models were used to analyze the hazard ratios (HR) and 95% CI for the association between cumulative BP and the risk of CVD among older adults. The restricted cubic spline function was employed to examine the potential dose-response pattern between cumulative BP and the risk of CVD. Results:During a total of 40 230 person-years of follow-up, with a mean follow-up duration of (8.98±2.16) years, 768 new cases of CVD were identified. The older adults in the highest quartile (Q4) of cumulative SBP, DBP, and PP had a significantly higher risk of CVD compared to those in the lowest quartile (Q1), with a HR of 1.68 (95% CI: 1.36-2.06), 1.67 (95% CI: 1.36-2.06), and 1.52 (95% CI: 1.24-1.84), respectively (all P<0.001). For every 10 mmHg (1 mmHg=0.133 kPa)×year increase in cumulative SBP, DBP, and PP, the risk of CVD increased by 2% ( HR=1.02, 95% CI: 1.01-1.03), 3% ( HR=1.03, 95% CI: 1.01-1.05), and 2% ( HR=1.02, 95% CI: 1.01-1.03), respectively (all P<0.05). The restricted cubic spline model revealed a linear positive correlation between cumulative SBP, DBP, and PP with the risk of CVD among older adults (all P for nonlinearity>0.05). When cumulative SBP, DBP, and PP exceeded 791.9 mmHg×years, 462.9 mmHg×years, and 323.6 mmHg×years, corresponding to an average BP level of 132.0 mmHg, 77.2 mmHg, and 53.9 mmHg, respectively, the risk of CVD began to increase. Conclusions:Elevated long-term cumulative BP may increase the risk of CVD in older adults. Caution is warranted when SBP, DBP, and PP exceed 132.0 mmHg, 77.2 mmHg, and 53.9 mmHg, respectively.

Objective To investigate the role and clinical significance of Calpain activity and 145 kDa cleavage fragments of αⅡ-spec-trin breakdown products(SBDP145)in systemic lupus erythematosus(SLE).Methods 124 patients with confirmed SLE and de-tailed clinical data were collected as the SLE group,and 75 healthy individuals who underwent physical examination during the same period were selected as the control group.Their serum samples were collected,and serum SBDP145 levels and Calpain activity were de-tected by the enzyme-linked immunosorbent assay(ELISA)and substrate-based method,respectively.The Mann-Whitney U test,Spearman rank correlation,univariate and multivariate Logistic regression,and receiver operating characteristic(ROC)curve were used to evaluate the clinical value of Calpain activity and SBDP145 in the diagnosis and assessment of SLE.Results Compared with healthy controls,serum Calpain activity([17.000[8.5000,33.500]RFU vs 7.500[4.000,12.500]RFU)and SBDP145 levels(5.283[3.532,9.463]ng/mL vs 1.472[0.994,2.212]ng/mL)in SLE patients were significantly increased(Z=-9.229,P<0.001;Z=-6.881,P<0.001).Furthermore,the Calpain activity was significantly correlated with SLE disease activity index(SLEDAI-2K)score(r=0.349,P<0.001).Logistic regression analysis showed that the increased Calpain activity and SBDP145 levels were significantly associated with the occurrence of SLE(OR=1.164,95％CI:1.016-1.334,P=0.029;OR=3.822,95％CI:1.928-7.574,P<0.001).The ROC curve analysis showed that the area under the ROC curve(AUCROC)of serum Calpain activity in distin-guishing SLE and healthy controls was 0.762(95％CI:0.697-0.827).When the cut-off value was 13.75 RFU,its sensitivity and speci-ficity were 63.1％and 81.1％,respectively.The AUCROC of serum SBDP145 levels in distinguishing SLE and healthy controls was 0.891(95％CI:0.845-0.936).When the cut-off value was 2.377 ng/mL,its sensitivity and specificity were 88.7％and 80.0％,re-spectively.The AUCROC,sensitivity,and specificity of combining the two with traditional SLE clinical indicators,including complement C3,erythrocyte sedimentation rate,and anti-dsDNA antibodies,in the diagnosis of SLE reached 0.986(95％CI:0.972-1.000),93.3％,and 96.0％,respectively.The Spearman correlation analysis results showed that the organ damage index(SDI)score of SLE patients was positively correlated with serum Calpain activity and SBDP145 levels(r=0.342,P<0.001;r=0.250,P=0.005).Con-clusion The elevated Calpain activity and SBDP145 levels are closely related to the occurrence and development of SLE,suggesting that they may be served as potential biomarkers for the diagnosis and assessment of SLE patients.

Objective:To investigate the role and mechanism of leucine-rich α-2-glycoprotein 1 (LRG1) in the fibrosis of human pterygium fibroblasts (HPFs).Methods:A total of 30 nasal primary pterygium tissues from patients who underwent pterygium excision surgery and 30 nasal normal conjunctival tissues from patients who underwent strabismus correction surgery were collected from the First Affiliated Hospital of Harbin Medical University between January 2022 and March 2023, serving as the pterygium group and normal control group, respectively.LRG1 protein expression in both groups was detected by immunofluorescence staining.The mRNA and protein levels of LRG1 and transforming growth factor-β1 (TGF-β1) were evaluated by quantitative real-time PCR (qRT-PCR) and Western blot.Primary HPFs were cultured from excised pterygium tissues using tissue block adhesion method, and cell morphology was observed.Vmentin and cytokeratin were identified by immunofluorescence staining.HPFs were divided into recombinant human LRG1 (rhLRG1) group and blank control group treated with or without 10 μg/ml rhLRG1 for 24 hours, respectively, and cell migration was evaluated via scratch assay.Additionally, HPFs were divided into blank control group, LRG1 overexpression group and LRG1 knockdown group.HPFs in LRG1 overexpression group and LRG1 knockdown group were transfected with LRG1 overexpression plasmids and small interfering RNA for 24 hours, respectively.TGF-β1 mRNA level was evaluated by qRT-PCR and expression of TGF-β1, fibronectin (FN), type Ⅲ collagen (COL3), and α-smooth muscle actin (α-SMA) proteins were evaluated by Western blot.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of the First Affiliated Hospital of Harbin Medical University (No.2022IIT026).Written informed consent was obtained from each subject.Results:HPFs were successfully isolated, exhibiting spindle-shaped morphology with whorled arrangement, positive identification for vimentin, and negative immunofluorescence staining for cytokeratin.The migration rate of the rhLRG1 group was (83.01±2.56)%, significantly higher than (50.32±4.97)% of the blank control group ( t=9.59, P<0.001).Immunofluorescence staining results showed that compared with normal conjunctival tissue, LRG1 protein was significantly higher expressed in pterygium tissue and was widely distributed in fibrous connective tissue and epithelial layer.Both mRNA and protein levels of LRG1 and TGF-β1 were significantly higher in the pterygium group than in the normal control group (mRNA: t=10.18, 6.15, both P<0.05.protein: t=6.83, 8.79, both P<0.05).In the LRG1 overexpression group, mRNA level of TGF-β1, and protein levels of FN, COL3 and α-SMA were significantly increased compared with the blank control and LRG1 knockdown groups (all P<0.05). Conclusions:LRG1 promotes fibrosis and enhances the migration ability in HPFs, and its mechanism may be associated with the upregulation of the TGF-β1 signaling pathway.

Objective:To analyze the prevalence and influencing factors of arteriosclerosis in different examination items in health check-up population.Methods:It was a cross-sectional study. A total of 4 479 subjects who underwent fundus, carotid artery color ultrasound and arteriosclerosis detector examinations in the Health Management Center at Tianjin Medical University General Hospital from January to December 2019 were selected as the study objects. The data including age, gender, family history and biochemical indicators were collected. The detection outcomes of arteriosclerosis and the distribution in different age and gender subgroups were analyzed. Kappa consistency test was used to analyze the consistency of different examination items of arteriosclerosis examination. The multiple logistic regression model was used to analyze the influencing factors of different examination items of arteriosclerosis detection. Results:Among the 4 479 subjects included in the analysis, 2 450 were male and 2 029 were female, with a mean age of (44.48±8.12) years. Arteriosclerosis was detected in one examination item in 1 167 cases (26.05%), 1 042 cases (23.26%) in two examination items, and 617 cases (13.78%) in three examination items, respectively. Among patients aged ≤40 years, 149 cases (18.91%) were detected with arteriosclerosis. In the age group of 40<-60 years, arteriosclerosis was detected in 2 660 patients (72.40%). All patients >60 years old exhibited arteriosclerosis, with 16 cases (94.12%) showing ≥2 items of arteriosclerosis. The results of fundus and carotid artery color Doppler ultrasound, fundus and arteriosclerosis detector, carotid artery color Doppler ultrasound and arteriosclerosis detector all demonstrated moderate consistency ( Kappa=0.32, 0.34, 0.24; all P<0.01). Logistic regression analysis revealed that the age ( OR=2.07, 95% CI: 1.97-2.17), male ( OR=1.60, 95% CI: 1.14-2.23), body mass index ( OR=1.10, 95% CI: 1.05-1.15), and systolic blood pressure ( OR=1.02, 95% CI: 1.01-1.03) were positively correlated with fundus arteriosclerosis (all P<0.05). Similarly, age ( OR=1.13, 95% CI: 1.12-1.14), male ( OR=2.53, 95% CI: 2.07-3.09), systolic blood pressure ( OR=1.02, 95% CI: 1.01-1.03), and low-density lipoprotein cholesterol ( OR=1.61, 95% CI: 1.16-2.23) were positively correlated with cervical arteriosclerosis, while high-density lipoprotein cholesterol ( OR=0.48, 95% CI: 0.32-0.72) was negatively correlated with cervical arteriosclerosis (all P<0.05). Furthermore, age ( OR=1.09, 95% CI: 1.08-1.11), male ( OR=1.55, 95% CI: 1.25-1.92), systolic blood pressure ( OR=1.08, 95% CI: 1.07-1.09), diastolic blood pressure ( OR=1.02, 95% CI: 1.00-1.03), postprandial blood glucose ( OR=1.09, 95% CI: 1.03-1.15), homocysteine ( OR=1.02, 95% CI: 1.00-1.03), and fibrinogen ( OR=1.26, 95% CI: 1.04-1.52) were positively correlated with systemic arterial stiffness (all P<0.05). Conclusions:The detection rates of arteriosclerosis in different examination items among the health check-up population vary. Age, male gender and systolic blood pressure are positive correlation factors for the three detection items of arteriosclerosis. For the examinees with aggregated risk factors, the multi-site and combined screening mode has certain significance for the early diagnosis and screening of arteriosclerotic diseases.

Objective:To explore the association between cumulative blood pressure (BP) and the risk of cerebrovascular disease (CVD) in older adults.Methods:This retrospective cohort study consecutively selected 4 480 older adults who participated in the Chinese Longitudinal Healthy Longevity Survey from 2008 to 2009 as the study subjects. The cumulative BP was calculated using the area under the curve from measurements taken at baseline (2008-2009), the first follow-up (2011), and the second follow-up (2014). The subjects were grouped with the quartiles (Q1-Q4) of cumulative systolic blood pressure (SBP), diastolic blood pressure (DBP), and pulse pressure (PP). The Cox proportional hazards regression models were used to analyze the hazard ratios (HR) and 95% CI for the association between cumulative BP and the risk of CVD among older adults. The restricted cubic spline function was employed to examine the potential dose-response pattern between cumulative BP and the risk of CVD. Results:During a total of 40 230 person-years of follow-up, with a mean follow-up duration of (8.98±2.16) years, 768 new cases of CVD were identified. The older adults in the highest quartile (Q4) of cumulative SBP, DBP, and PP had a significantly higher risk of CVD compared to those in the lowest quartile (Q1), with a HR of 1.68 (95% CI: 1.36-2.06), 1.67 (95% CI: 1.36-2.06), and 1.52 (95% CI: 1.24-1.84), respectively (all P<0.001). For every 10 mmHg (1 mmHg=0.133 kPa)×year increase in cumulative SBP, DBP, and PP, the risk of CVD increased by 2% ( HR=1.02, 95% CI: 1.01-1.03), 3% ( HR=1.03, 95% CI: 1.01-1.05), and 2% ( HR=1.02, 95% CI: 1.01-1.03), respectively (all P<0.05). The restricted cubic spline model revealed a linear positive correlation between cumulative SBP, DBP, and PP with the risk of CVD among older adults (all P for nonlinearity>0.05). When cumulative SBP, DBP, and PP exceeded 791.9 mmHg×years, 462.9 mmHg×years, and 323.6 mmHg×years, corresponding to an average BP level of 132.0 mmHg, 77.2 mmHg, and 53.9 mmHg, respectively, the risk of CVD began to increase. Conclusions:Elevated long-term cumulative BP may increase the risk of CVD in older adults. Caution is warranted when SBP, DBP, and PP exceed 132.0 mmHg, 77.2 mmHg, and 53.9 mmHg, respectively.

Tumor necrosis factor-α(TNF-α)is involved in the regulation of multiple biological processes such as the proliferation,invasion,migration,and chemotherapy resistance of hepatocellular carcinoma(HCC)cells through TNF receptor-mediated signaling pathways.At the same time,TNF-α also plays a role in inducing the apoptosis of HCC cells.Some TNF-α inhibitors have been shown to inhibit the progression of HCC and prolong survival time.At present,the potential mechanism of action of TNF-α in HCC remains unclear,and exploration of the interaction between TNF-α and HCC can help to determine the potential therapeutic targets for HCC.This article summarizes the latest research advances in the mechanism of action of TNF-α in HCC and introduces the possibility of targeting TNF-α as a treatment method for HCC,in order to provide a theoretical basis for the prevention and treatment of liver cancer and drug research and development.

Objective:To investigate the effect and mechanism of isorhynchophylline (IRN) on angiotensin Ⅱ(Ang Ⅱ)-induced cardiac hypertrophy.Methods:H9c2 cells were co-cultured with Ang Ⅱ and different concentrations of IRN (0, 5, 10, 25, 50 μmol/L). The cell surface area and mRNA levels of cardiac hypertrophy markers atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and β-myosin heavy chain (β-MHC) were detected to elucidate the effect of IRN on myocardial hypertrophy and the most effective concentration. H9c2 cells were co-cultured with Ang Ⅱ and IRN (25 μmol/L) at different times (0, 6, 12, 24 h) to elucidate the most effective time of inhibition. The phosphorylation levels of the signaling pathway were detected, and the effects of IRN and Akt inhibitor MK2206 on the phosphorylation levels of the signaling pathway were further explored to elucidate the underlying mechanisms.Results:Compared with the control group, the surface area of H9c2 cells, and the mRNA expression of myocardial hypertrophy markers ANP, BNP and β-MHC were significantly increased (all P<0.05). Pretreated with different concentrations of IRN (5, 10, 25, 50 μmol/L) could inhibit the increase in cell surface area induced by AngⅡ (all P<0.05), especially at the concentration of 25 μmol/ L ( P<0.01). IRN could time-dependently inhibit AngⅡ-induced activation of ANP, BNP, β-MHC mRNA (all P<0.05). AngⅡ caused increased phosphorylation levels of Akt, GSK3β, mTOR and FOXO3a. IRN could block AngⅡ-induced phosphorylation of the Akt signaling pathway. Conclusion:IRN attenuates AngⅡ-induced cardiomyocyte hypertrophy by inhibiting the Akt signaling pathway.