Objective:To investigate the relationship between different obesity metabolic phenotypes and the incidence of new carotid artery plaque.Methods:The present study is a retrospective cohort study, collecting individuals from the Health Management Center of the Second Affiliated Hospital of Dalian Medical University who had two or more cervical vascular color ultrasound examinations and met the inclusion criteria from 2014 to 2022, and collected their baseline clinical data. According to whether the subjects were obese and had metabolic syndrome, they were divided into metabolically healthy non-obese group, metabolically unhealthy non-obese group, metabolically healthy obese group, and metabolically unhealthy obese group. The first physical examination time of the subjects was taken as the starting point of follow-up, and cervical vascular color ultrasound was performed during the follow-up physical examination, with the outcome event being carotid artery plaque. Kaplan-Meier survival curve analysis was used to analyze the cumulative incidence of carotid artery plaques in the four groups and log-rank test was performed, and a multifactorial Cox proportional hazards model was used to analyze the relationship between different obesity metabolic phenotypes and the risk of carotid artery plaque incidence.Results:A total of 4 890 subjects were enrolled, aged (45.4±9.6) years, and 2 754 (56.3%) males. The follow-up time was 1.14(0.93, 2.20) years. Compared with the other 3 obesity metabolic phenotypes, the incidence of carotid plaques in the metabolically unhealthy obesity group was the highest (15.4% (286/1 861)). Kaplan-Meier survival curve analysis showed that the cumulative incidence of carotid plaques in metabolically unhealthy obese subjects was about 2.962 times that of metabolically healthy non-obese subjects (log-rank P<0.001). Multivariate Cox regression results showed that the risk of carotid plaque in metabolically unhealthy obese subjects was 1.650 times that of metabolically healthy non-obese subjects (95% CI: 1.203-2.264, P=0.002). Conclusion:Metabolically unhealthy obesity phenotype is an independent risk factor for carotid plaque.

Objective:To construct a novel dual-specific antibody targeting human CD123 (CD123 DuAb) and study its effects in acute myeloid leukemia (AML) .Methods:Based on the variable region of the CD123 monoclonal antibody independently developed at our institution, the CD123 DuAb expression plasmid was constructed by molecular cloning and transfected into ExpiCHO-S cells to prepare the antibody protein. Through a series of in vitro experiments, its activation and proliferation effect on T cells, as well as the effect of promoting T-cell killing of AML cells, were verified.Results:① A novel CD123 DuAb plasmid targeting CD123 was successfully constructed and expressed in the Expi-CHO eukaryotic system. ②The CD123 DuAb could bind both CD3 on T cells and CD123 on CD123 + tumor cells. ③When T cells were co-cultured with MV4-11 cells with addition of the CD123 DuAb at a concentration of 1 nmol/L, the positive expression rates of CD69 and CD25 on T cells were 68.0% and 44.3%, respectively, which were significantly higher than those of the control group ( P<0.05). ④Co-culture with CD123 DuAb at 1 nmol/L promoted T-cell proliferation, and the absolute T-cell count increased from 5×10 5/ml to 3.2×10 6/ml on day 9, and CFSE fluorescence intensity decreased significantly. ⑤ With the increase in CD123 DuAb concentration in the culture system, T-cell exhaustion and apoptosis increased. When the CD123 DuAb was added at a concentration of 1 nmol/L to the culture system, the proportion of CD8 + PD-1 + LAG-3 + T cells was 10.90%, and the proportion of propidium iodide (PI) - Annexin Ⅴ + T cells and PI + Annexin Ⅴ + T cells was 18.27% and 11.43%, respectively, which were significantly higher than those in the control group ( P<0.05). ⑥ The CD123 DuAb significantly activated T cells, and the activation intensity was positively correlated with its concentration. The expression rate of CD107a on T cells reached 16.05% with 1 nmol/L CD123 DuAb, which was significantly higher than that of the control group ( P<0.05). ⑦The CD123 DuAb promoted cytokine secretion by T cells at a concentration of 1 nmol/L, and the concentration of IFN-γ and TNF-α in the supernatant of the co-culture system reached 193.8 pg/ml and 169.8 pg/ml, respectively, which was significantly higher than that of the control group ( P<0.05). ⑧When CD123 DuAb was added at a concentration of 1 nmol/L to the co-culture system of T cells and CD123 + tumor cells, the killing intensity of T cells significantly increased, and the residual rates of CD123 + MV4-11 cells, CD123 + Molm13 cells, and CD123 + THP-1 cells were 7.4%, 6.7%, and 14.6% on day 3, respectively, which were significantly lower than those in the control group ( P<0.05) . Conclusion:In this study, a novel CD123 DuAb was constructed and expressed. In vitro experiments verified that the DuAb binds to CD123 + tumor cells and T cells simultaneously, promotes T-cell activation and proliferation, and facilitates their anti-leukemia effect, which provides a basis for further clinical research.

Objective:To evaluate the efficacy and safety of hypomethylating agents (HMA) in patients with myelodysplastic syndromes (MDS) .Methods:A total of 409 MDS patients from 45 hospitals in Zhejiang province who received at least four consecutive cycles of HMA monotherapy as initial therapy were enrolled to evaluate the efficacy and safety of HMA. Mann-Whitney U or Chi-square tests were used to compare the differences in the clinical data. Logistic regression and Cox regression were used to analyze the factors affecting efficacy and survival. Kaplan-Meier was used for survival analysis. Results:Patients received HMA treatment for a median of 6 cycles (range, 4-25 cycles) . The complete remission (CR) rate was 33.98% and the overall response rate (ORR) was 77.02%. Multivariate analysis revealed that complex karyotype ( P=0.02, OR=0.39, 95% CI 0.18-0.84) was an independent favorable factor for CR rate. TP53 mutation ( P=0.02, OR=0.22, 95% CI 0.06-0.77) was a predictive factor for a higher ORR. The median OS for the HMA-treated patients was 25.67 (95% CI 21.14-30.19) months. HMA response ( P=0.036, HR=0.47, 95% CI 0.23-0.95) was an independent favorable prognostic factor, whereas complex karyotype ( P=0.024, HR=2.14, 95% CI 1.10-4.15) , leukemia transformation ( P<0.001, HR=2.839, 95% CI 1.64-4.92) , and TP53 mutation ( P=0.012, HR=2.19, 95% CI 1.19-4.07) were independent adverse prognostic factors. There was no significant difference in efficacy and survival between the reduced and standard doses of HMA. The CR rate and ORR of MDS patients treated with decitabine and azacitidine were not significantly different. The median OS of patients treated with decitabine was longer compared with that of patients treated with azacitidine (29.53 months vs 20.17 months, P=0.007) . The incidence of bone marrow suppression and pneumonia in the decitabine group was higher compared with that in the azacitidine group. Conclusion:Continuous and regular use of appropriate doses of hypomethylating agents may benefit MDS patients to the greatest extent if it is tolerated.

Objective To investigate the role and mechanism of sine oculis homeobox 1(Six1),a nephrogenic transcription factor,in regulating injury repair after acute kidney injury(AKI).Methods C57BL/6J mice were inflicted with renal ischemia reperfusion(IR)injury to establish AKI model,and then the serum samples and kidney tissues were collected at days 1,2,and 3 after modeling.Serum creatinine(Cr)and blood urea nitrogen(BUN)levels were measured and renal morphology was observed with HE staining.The expression and distribution of nephrogenic molecules(Six1 and Pax2,etc.)and cell proliferation/migration genes(Cyclin A1,MMP9,etc.)were detected with RT-PCR,Western blotting,and immunofluorescence assay and immunohistochemical assay.The expression of apoptosis pathway(Bc12,Caspase3,etc.)and cell proliferation related molecules were evaluated in Six1 overexpression/knockout human epithelial cells(HK2)with CoCl2-induced injury.Additionally,after the adeno-associated viruses carrying Six1 overexpression vector were used to overexpress the molecule in the mice,the expression of Six1 and other related molecules were detected after IR injury modeling.Results After renal IR injury,Six1 was significantly activated in epithelial cells,the expression of nephrogenic and cell proliferation/migration molecules(Pax2,Cyclin A1,MMP9,etc.)was obviously up-regulated(P＜0.05),which was positively correlated with Six1 expression,while the proliferation/migration molecules(Ki67,MMP9)were also localized within the tubular epithelial cells.In cellular models of Six1 overexpression,the expression levels of nephrogenic and cell proliferation/migration molecules(Pax2,Cyclin A1,MMP9,etc.)were also notably up-regulated(P＜0.05).In the mice with renal overexpression of Six1,the nephrogenic molecules as well as anti-apoptotic ones(Six2,Pax2,Bc12,etc.)were up-regulated,while the expression of kidney injury-related molecule(Kim1)in kidneys was reduced in the renal tissues.While,in 2 d after IR injury,the anti-apoptotic gene(Bc12,Stat3)was significantly up-regulated(P＜0.05),and the apoptotic and injury molecules(Kim1,Caspase3)showed remarkable down-regulation(P＜0.05)in the mice with renal Six1 overexpression.Furthermore,CoCl2-inducion significantly decreased the cell proliferation rate in the Six1 knockoutgroup(TCMK1-Six1-/-)(P＜0.05)but increased the rate in the Six1 overexpression group(TCMK1-Six1)when compared to the control cells(P＜0.05).And,the expression of nephrogenic,anti-apoptotic pathways and cell proliferation/migration molecules(Pax2,Bc12,Cyclin A1,MMP9,etc.)were reduced in TCMK1-Six1-/-group,and apoptosis and kidney injury molecules(Caspase3,Kim1)were significantly down-regulated in TCMK1-Six1 group(P＜0.05).Conclusion Activation of Six1 after AKI can promote the proliferation/migration of renal tubular epithelial cells by up-regulating nephrogenic molecules and inducing anti-apoptotic pathway molecules,and then,participate in IR-induced renal injury repair.

Objective:To investigate the association between different dietary patterns and subtypes of metabolic associated fatty liver disease(MAFLD).Methods:A total of 6 022 check-ups at the health management center of the Second Hospital of Dalian Medical University from January 2022 to March 2023 were selected as study subjects. MAFLD was categorised into three subtypes: overweight/obese type, metabolic disorder type, and diabetic type. Factor analysis was used to extract dietary patterns. Logistic regression was used to assess the impact of dietary patterns on MAFLD occurrence, constructing interaction models between dietary patterns intake and age, gender, and physical exercise levels. Results:Four dietary patterns were extracted based on feature sorting after factor analysis and were named as the high-quality protein pattern, the fruit-vegetable pattern, egg-aquatic pattern, and the processed meat pattern. Regression analysis of the unadjusted model showed that overweight/obese and diabetic types of MAFLD were negatively associated with the high-quality protein mode, while model-adjusted regression analysis showed that the processed meat pattern was positively associated with the risk of MAFLD, and fruit-vegetable pattern was positively associated with overweight/obese MAFLD( P<0.05). The results of subgroup analyses suggested that female( OR=1.55, 95% CI 1.14-2.15) with a high intake of pickle pattern had a higher risk of overweight/obese MAFLD than male( OR=1.18, 95% CI 1.02-1.49). Conclusion:High-quality protein pattern was negatively correlated with MAFLD, whereas fruit-vegetable pattern and processed meat pattern were positively correlated with MAFLD. Female with high consumption of processed meat pattern are more likely to develop overweight/obesity MAFLD compared with male. It is recommended that people with MAFLD reduce their intake of processed products and high-fructose food, and consume adequate amounts of high-quality protein food to maintain a balanced diet.

Objective:To investigate the relationship between obesity and incident chronic kidney disease(CKD) in a population undergoing health check-ups.Methods:This is a retrospective cohort study. A total of 31 251 participants who had at least 2 health physical examinations in the Health Management Center of the Second Affiliated Hospital of Dalian Medical University from January 2017 to December 2022 and met the inclusion criteria were selected. The participants were divided into normal body weight group, overweight group, and obese group according to baseline body mass index. Cox proportional hazard regression model was used to analyze the relationship between obesity and new-onset CKD, and the dose-response relationship between body mass index and CKD was analyzed with restricted cubic splines.Results:Multivariate Cox regression analysis showed that the risk of developing CKD increased by 13%( HR=1.13, 95% CI 1.01-1.25) and 55%( HR=1.55, 95% CI 1.36-1.76) in the overweight and obese group compared to the normal weight group. Subgroup analysis indicated that obese women had a higher risk of developing CKD compared to men. There was a " U-shaped" correlation between body mass index and CKD in male population, with the lowest risk of CKD occurring at body mass index of 19.6-24.2 kg/m 2. In women, the relationship between body mass index and CKD was approximately linear, with the risk of CKD gradually increasing when body mass index exceeded 22.5 kg/m 2. Conclusions:Obesity is an independent risk factor for new-onset CKD, and obese women have a higher risk of developing CKD than men. Regarding CKD prevention, men are advised to maintain a higher level of body weight within the normal range of body mass index, while women are encouraged to control their weight to a lower level within the normal body mass index range.

Objective:To explore the correlation between systemic immunity-inflammation index(SII) and hyperuricemia(HUA).Methods:Participants who had at least 3 health checkups in the Health Management Center of the Second Affiliated Hospital of Dalian Medical University from January 2014 to December 2022 were selected to construct a dynamic cohort. The SII, reflecting the inflammatory state of the body, was constructed using neutrophil, platelet, and lymphocyte counts. A Cox proportional hazard regression model was used to explore the association between SII and HUA in the overall population and different subgroups of the population, and sensitivity analysis was performed twice. Results:A total of 20 022 subjects were included, and the mean follow-up time was 3.67 years. After adjusting for confounding factors, each unit increase in the natural logarithm of SII(lnSII) was associated with a 24% increased risk of hyperuricemia( HR=1.24, 95% CI 1.16-1.32, P<0.001). As a categorical variable, compared with the lowest quartile array( Q1), the risk of HUA in the total population increased by 12%( HR=1.12, 95% CI 1.03-1.21, P=0.006), 14%( HR=1.14, 95% CI 1.06-1.24, P=0.001), 27%( HR=1.27, 95% CI 1.17-1.37, P<0.001) in Q2, Q3 and Q4 groups within the general population, respectively. All subgroup analysis and sensitivity analysis showed that SII was positively correlated with HUA. Conclusions:Elevated levels of SII significantly increase the risk of HUA. Assessing the body′s inflammatory status using SII can aid in risk screening and preventive management for individuals at high risk of HUA.

Objective To investigate the correlations of serum Apelin-13 and fatty acid binding protein 4 (FABP4) levels with metabolic and bone metabolic indicators in postmenopausal women with different bone mass. Methods A total of 145 postmenopausal women were selected as subjects and divided into three groups based on bone mineral density (BMD) test results: normal bone mass group(49 cases), osteopenia (ON) group(51 cases), and osteoporosis (OP) group(45 cases). Serum Apelin-13, FABP4 levels, bone metabolic indicators, and biochemical indicators were measured and compared among the three groups. Spearman correlation analysis was used to analyze the correlations of Apelin-13, FABP4, and other indicators with BMD. Multivariate Logistic regression analysis was performed to analyze the risk factors for OP, and the receiver operating characteristic (ROC) curve was plotted to analyze the predictive value of serum Apelin-13 for postmenopausal osteoporosis (PMOP). Results The serum Apelin-13 level in the OP group was lower than that in the ON group and the normal bone mass group (P < 0.05). No significant difference in serum FABP4 levels was found among the three groups (P>0.05). The levels of serum parathyroid hormone (PTH), alkaline phosphatase (ALP), type Ⅰ procollagen amino-terminal propeptide (PⅠNP), type Ⅰ collagen cross-linked C-terminal peptide (CTXⅠ), and bone-specific alkaline phosphatase (BALP) in the OP group were higher than those in the ON group and the normal bone mass group (P < 0.05). Lumbar post-menopause BMD was positively correlated with serum Apelin-13 levels (P < 0.05), but had no correlation with serum FABP4 levels (P>0.05). Lumbar BMD was negatively correlated with serum PTH, ALP, PⅠNP, CTXⅠ, BALP, and age (P < 0.05), but positively correlated with body weight, body mass index, T-score, fasting insulin, and insulin resistance index (P < 0.05). Multivariate Logistic regression analysis showed that serum Apelin-13, PTH, ALP, PⅠNP, CTXⅠ, and BALP levels were independent factors influencing the occurrence of OP in postmenopausal women (P < 0.05). ROC curve results showed that the optimal cut-off value of serum Apelin-13 for predicting PMOP was 18.51 pg/mL, with an area under the curve of 0.716, a sensitivity of 70.0%, and a specificity of 64.4%. Conclusion Apelin-13 is lowly expressed in the serum of PMOP patients, and its expression level is closely related to lumbar BMD, which may serve as an early screening indicator and potential therapeutic target for PMOP.

Objective:To establish a method for determintation of chlorogenic acid and linarin in Yejuhua granules by HPLC.Methods:We applied HPLC methods. The Kromasil 100-5 C18 column (250 mm×4.6 mm,5 μm) was used, the mobile phase was acetonitrile-0.4%H 3PO 4 solution (gradient elution), the flow rate was 1.0 ml/min, the dection wavelenghth was 334 nm and the column temperture was 32 ℃. Results:Chlorogenic acid and buddleoside had good linearity in the ranges of 0.30-1.50 μg ( r2=0.999 1) and 0.12-0.62 μg ( r2=0.999 8), respectively. The average recoveries were 99.70% and 96.67%, with RSD<2%, respectively. Conclusion:The method is simple, rapid, reliable, efficient, and can be used for determination of chlorogenic acid and buddleoside in Yejuhua Granules.

Exposure to particulate matter 2.5 (PM2.5) potentially triggers airway inflammation by activating nuclear factor-κB (NF-κB). Sirtuin 2 (SIRT2) is a key modulator in inflammation. However, the function and specific mechanisms of SIRT2 in PM2.5-induced airway inflammation are largely understudied. Therefore, this work investigated the mechanisms of SIRT2 in regulating the phosphorylation and acetylation of p65 influenced by PM2.5-induced airway inflammation and bronchial hyperresponsiveness. Results revealed that PM2.5 exposure lowered the expression and activity of SIRT2 in bronchial tissues. Subsequently, SIRT2 impairment promoted the phosphorylation and acetylation of p65 and activated the NF-κB signaling pathway. The activation of p65 triggered airway inflammation, increment of mucus secretion by goblet cells, and acceleration of tracheal stenosis. Meanwhile, p65 phosphorylation and acetylation, airway inflammation, and bronchial hyperresponsiveness were deteriorated in SIRT2 knockout mice exposed to PM2.5. Triptolide (a specific p65 inhibitor) reversed p65 activation and ameliorated PM2.5-induced airway inflammation and bronchial hyperresponsiveness. Our findings provide novel insights into the molecular mechanisms underlying the toxicity of PM2.5 exposure. Triptolide inhibition of p65 phosphorylation and acetylation could be an effective therapeutic approach in averting PM2.5-induced airway inflammation and bronchial hyperresponsiveness.
Animals ; Inflammation ; Mice ; NF-kappa B/metabolism* ; Particulate Matter/toxicity* ; Signal Transduction ; Sirtuin 2/metabolism* ; Transcription Factor RelA/metabolism*