Objective This study aims to investigate the therapeutic effect and mechanism of Naoxinqing on non-alcohol fatty liver disease (NAFLD) induced by a high-fat diet through network pharmacology,molecular docking and in vitro and in vivo experiments. Methods ApoE-/-mice were given a high-fat diet for 12 weeks to establish the NAFLD model,followed by a 12-week Naoxinqing administration. To evaluate the therapeutic effect of Naoxinqing on NAFLD induced by a high-fat diet,biochemical and histopathological experiments were performed,including assessment of blood lipids,liver function,serum inflammatory factors,as well as Hematoxylin and eosin (HE),Oil red O,and Sirius red staining of liver. Subsequently,network pharmacology and molecular docking techniques were employed to predict the key targets of Naoxinqing. Finally,the mechanism of Naoxinqing was validated by Western Blot in HepG2 cells and liver tissue. Results The results of serum biochemistry and liver tissue pathology showed that Naoxinqing can significantly improve high-fat diet-induced hepatic lipid accumulation,hepatocellular injury,and inflammation. Network pharmacology and molecular docking analysis results suggested that Naoxinqing may affect lipid metabolism through the AMP-activated protein kinase (AMPK)/Sirtuin 1 (SIRT1) pathway. Finally,in vitro cell experiment confirmed that the main mechanism of Naoxinqing is to activative the AMPK/SIRT1 pathway,upregulate the expression of downstream carnitine palmitoyltransferase 1 (CPT1A),promote fatty acid oxidation,and ultimately improve NAFLD. Conclusion This study demonstrated that Naoxinqing improved NAFLD by promoting fatty acid oxidation through the activation of the AMPK/SIRT1 pathway.

Increasing evidences suggest the important role of calcium homeostasis in hallmarks of cancer, but its function and regulatory network in metastasis remain unclear. A comprehensive investigation of key regulators in cancer metastasis is urgently needed. Transcriptome sequencing (RNA-seq) of primary esophageal squamous cell carcinoma (ESCC) and matched metastatic tissues and a series of gain/loss-of-function experiments identified potassium channel tetramerization domain containing 4 (KCTD4) as a driver of cancer metastasis. KCTD4 expression was found upregulated in metastatic ESCC. High KCTD4 expression is associated with poor prognosis in patients with ESCC and contributes to cancer metastasis in vitro and in vivo. Mechanistically, KCTD4 binds to CLIC1 and disrupts its dimerization, thus increasing intracellular Ca²⁺ level to enhance NFATc1-dependent fibronectin transcription. KCTD4-induced fibronectin secretion activates fibroblasts in a paracrine manner, which in turn promotes cancer cell invasion via MMP24 signaling as positive feedback. Furthermore, a lead compound K279-0738 significantly suppresses cancer metastasis by targeting the KCTD4‒CLIC1 interaction, providing a potential therapeutic strategy. Taken together, our study not only uncovers KCTD4 as a regulator of calcium homeostasis, but also reveals KCTD4/CLIC1-Ca²⁺-NFATc1-fibronectin signaling as a novel mechanism of cancer metastasis. These findings validate KCTD4 as a potential prognostic biomarker and therapeutic target for ESCC.

Background Individual lead or cadmium exposure can cause abnormal blood glucose level and changes in telomere length, and the role of telomere length in the relationship between heavy metal joint exposure and blood glucose level is still unclear. Objective To explore the role of telomere length in the relationship between lead and cadmium coexposure and blood glucose. Methods A cross-sectional study was conducted. By convenient sampling method, 600 residents living in two communities in a city in North China were selected as participants from April to June 2016. Face-to-face interviews were performed to collect general demographics and lifestyles of the participants. The peripheral blood samples of the participants were collected for blood glucose and telomere length detection, the urine samples were collected for urinary cadmium, urinary lead, and urinary creatinine measurement, and both urinary cadmium and urinary lead were corrected by urinary creatinine. The included participants were divided into a control group, a high-cadmium and low-lead group, a high-lead and low-cadmium group, and a high-lead and high-cadmium group, according to the median levels of urinary cadmium and urinary lead. A restricted cubic spline model was constructed to analyze the relationship between urinary lead/cadmium levels and blood glucose concentrations in the four groups and the relationship between cadmium exposure and telomere length in the high-lead and high-cadmium group. Intermediary model test was conducted to analyze the effect of telomere length on the relationship between exposures to lead and cadmium and blood glucose. Results The included participants were divided into the control group (n=99), the high-cadmium and low-lead group (n=91), the high-lead and low-cadmium group (n=145), and the high-lead and high-cadmium group (n=265). The differences in age, education level, per capita monthly household income, smoking, blood glucose, and telomere length were statistically significant among the four groups (P＜0.05). The high-lead and high-cadmium group had the highest blood glucose concentration, (5.63±1.68) mmol·L⁻¹, and the shortest telomere length, (2.63±1.05) Kb. The restricted cubic spline results showed that urinary cadmium level was correlated with blood glucose concentration in the high-lead and high-cadmium group (F=3.45, P=0.037), and there was a non-linear association (F=6.91, P=0.002); the association between urinary cadmium level and telomere length was also non-linear (F=5.93, P=0.043). The intermediary model test results showed that telomere length was a mediating variable between urinary cadmium level and blood glucose concentration, and the mediating effect size was 0.0192 (95%CI: 0.0007-0.0563), with a mediation ratio of 15.57%. Conclusion Correlations between urinary cadmium and blood glucose and between urinary cadmium and telomere length were observed in the high-lead and high-cadmium coexposure group, and telomere length may play a mediating role in the relationship between them.

Background Environmental pollutants can affect N⁶-methyladenosine (m⁶A) level in the body, but the change of m⁶A level in kidney after being exposed to cadmium (Cd) and the molecular mechanism of renal injury need to be further studied. Objective To analyze the associations of m⁶A modification and methyltransferases/demethylases with microRNA-21 (miR-21) and transforming growth factor- β1 (TGF - β1) in kidney of rats exposed to Cd. Methods Twenty-four SPF male SD rats were divided into 4 groups, with 6 rats in each group, and were exposed to Cd by subcutaneous injection of 2.0, 1.0, and 0.5 mg·kg⁻¹ cadmium chloride (CdCl₂) and equal volume of normal saline for 2 weeks, 7 d a week, respectively. The levels of N-acetyl-β-D-glucosidase (UNAG) and albumin (UALB) in urine, and the levels of m⁶A methylation and TGF-β1 in kidney were detected by enzyme-linked immunosorbent assay (ELISA). The level of blood urea nitrogen (BUN) was measured by urease method. The levels of renal oxidative stress indicators such as malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) were detected by total bile acid method, water-soluble tetrazolium asssay, and colorimetric method respectively. The relative levels of TGF-β1, methyltransferases, and demethylases in kidney were measured by reverse transcription-polymerase chain reaction. The expression of miR-21 in kidney was detected by fluorescent quantitative polymerase chain reaction. Results After 2 weeks of exposure to Cd, the body weights of rats in the 2.0 and 1.0 mg·kg⁻¹ cadmium chloride groups decreased, and the ratio of kidney/body weight and the levels of BUN, UNAG, and TGF-β1 mRNA and protein increased in the 2.0 mg·kg⁻¹ cadmium chloride group (P<0.05). The expression levels of m⁶A modification, methyltransferases METTL3, METTL14, Wilms’ tumor 1-associated protein (WTAP), and miR-21 were increased both in the 2.0 and 1.0 mg·kg⁻¹ cadmium chloride groups, with significant differences compared with the control group (P<0.05). The results of correlation analysis showed that the m⁶A modification level was negatively correlated with SOD (r=−0.4489, P<0.05) and GSH-Px (r=−0.4874, P<0.05), METTL3 was negatively correlated with MDA (r=−0.5158, P<0.05), while there was a positive correlation between FTO and GSH-Px (r=0.4802, P<0.05). In addition, miR-21 was positively correlated with METTL3 (r=0.7491), METTL14 (r=0.6157), and WTAP (r=0.6660) (P<0.05), TGF-β1 was positively correlated with METTL3 (r=0.5025, P<0.05) but negatively correlated with FTO (r=−0.5634, P<0.05) . Conclusion Cd can induce m⁶A methylation and up-regulation of METTL3, METTL14, WTAP, and miR-21 expression levels in rat kidney tissues, indicating that m⁶A and miR-21 may be associated with Cd-induced renal fibrosis.

Objective:To investigate the association between pulse pressure(PP) and new-onset diabetes in overweight and obese people.Methods:A prospective cohort study was conducted in overweight or obese participants selected from Kailuan Study who underwent 2006-2007 annual checkup and met the inclusion and exclusion criteria. PP was calculated by blood pressure and participants were divided into 4 groups according to PP quartile. The cumulative incidence of new-onset diabetes of different PP groups was calculated by Kaplan- Meier method and compare by Log- Rank test. The multivariate Cox proportional hazards model was used to analyze the association between different PP groups and new-onset diabetes. Results:During an average follow-up of 8.45 years, 8 922 diabetes was identified. The cumulative incidence rate of the Q1, Q2, Q3, and Q4 groups were 22.12%, 24.48%, 27.97%, and 33.44% respectively, which were statistically different( χ2=368.16, P<0.01). Cox proportional hazards regression analysis showed that after adjusting for multiple confounding factors, compared with Q1 group, the hazard ratio for diabetes in Q2, Q3, and Q4 groups were 1.07(1.00-1.14), 1.13(1.05-1.21), and 1.17(1.09-1.27) respectively. And the HR of diabetes event in pulse pressure(per 1 SD increase) was 1.04(1.02-1.07). Similar results were found in participants who were over-weight, obese, with normal blood pressure or hypertensive without drugs use. Conclusion:PP is positively correlated with the new-onset diabetes. High PP is one of the risk factors for developing diabetes in overweight and obese people.

Background Exposure to environmental lead can cause kidney damage and telomere wear. However, the relationship among lead, peripheral blood telomere length, and glomerular filtration rate (eGFR) are unclear. Objective This study is conducted to investigate the relationships of urinary lead level with peripheral blood telomere length and renal function index eGFR, and further explore whether peripheral blood telomere length plays an intermediary role in the relationship between urinary lead level and eGFR. Methods A case-control study was conducted to select 497 residents from two communities in a city, including 230 in the control group (eGFR≥80 mL·min⁻¹) and 267 in the abnormal eGFR group (eGFR<80 mL·min⁻¹). Basic information and health information of the subjects were collected through a face-to-face questionnaire survey. Fasting morning urine was collected, and urinary lead and urinary creatinine (UCr) were detected. Fasting peripheral venous blood was collected to detect telomere length and serum creatinine (SCr) in peripheral blood leukocytes. eGFR was estimated by the Levey formula. After further adjusting for age, gender, education level, family per capita monthly income, smoking, and drinking the relationship among urinary lead level, peripheral blood telomere length, and renal function index eGFR was evaluated by mediating effect analysis. Results The overall level of creatinine-adjusted urinary lead [M (P₂₅, P₇₅)] in the abnormal eGFR group was 3.85 (1.56, 7.34) μg·g⁻¹ which was higher than that in the control group, 1.57 (0.60, 3.62) μg·g⁻¹(P<0.001). In addition, the overall level of peripheral blood telomere length in the abnormal eGFR group was 2.42 (1.89, 3.10) Kb, lower than that in the control group, 2.69 (2.09, 3.64) Kb (P<0.001). The results of mediating effect analysis showed that the magnitude of mediating effect by peripheral blood telomere length was −0.276 (95%CI: −0.708-−0.001) and it contributed 3.35% to the relationship between urinary lead level and eGFR. In women, the magnitude of mediating effect by peripheral blood telomere length was −0.484 (95%CI: −1.160-−0.023) between urinary lead level and eGFR, and the proportion of the mediating effect was 5.34%. In men, no mediating role of peripheral blood telomere length was found between urinary lead and eGFR. Conclusion Urinary lead level is closely related to renal function index eGFR and telomere length in peripheral blood. Peripheral blood telomere length plays a mediating role in the relationship between female urinary lead and eGFR in women.

Objective:To investigate the value of serum urea nitrogen on in-hospital death in patients with heart failure.Methods:The clinical data of 9 459 patients with heart failure from January 2013 to December 2018 in Shengjing Hospital of China Medical University were retrospectively analyzed. Among them, 296 cases died in hospital (death group) and 9 163 cases survived (survival group). The clinical data of patients were collected, including general condition, disease history, physical examination, laboratory indicators and relevant physical examination, etc. Correlation was finished with Pearson correlation analysis. Multivariate Logistic regression analysis was used to determine independent risk factors for in-hospital death in patients with heart failure. Receiver operating characteristic (ROC) curve was used to determine the optimal predictive threshold of urea nitrogen for in-hospital death.Results:The in-hospital mortality in patients with heart failure was 3.1% (296/9 459). There were statistical differences in age, hypertension rate, diabetes rate, a history of atrial fibrillation rate, smoking history rate, hemoglobin, albumin, glycosylated hemoglobin, urea nitrogen, creatinine, uric acid, serum potassium, serum sodium, troponin I, N terminal brain natriuretic peptide precursor (NT-proBNP), left ventricular ejection fraction (LVEF) between death group and survival group ( P<0.01 or <0.05), and there were no statistical difference in gender composition, coronary heart disease rate, platelet, total cholesterol, low-density lipoprotein cholesterol (LDL-C), triglyceride, left ventricular end-diastolic volume (LVEDV) and left ventricular end-systolic volume (LVESV) between 2 groups ( P>0.05). Pearson correlation analysis result showed that the urea nitrogen was positively correlated with age, coronary heart disease, hypertension, diabetes, glycosylated hemoglobin, creatinine, uric acid, serum potassium, troponin I, NT-proBNP, LVEDV and LVESV ( r = 0.130, 0.024, 0.053, 0.128, 0.033, 0.739, 0.468, 0.377, 0.065, 0.432, 0.084 and 0.101; P<0.01 or <0.05); and the urea nitrogen was negatively correlated with gender, history of atrial fibrillation, hemoglobin, platelet, albumin, total cholesterol, LDL-C, serum sodium and LVEF ( r = -0.033, -0.063, -0.272, -0.077, -0.188, -0.070, -0.071, -0.199 and -0.113, P<0.01); and there were no correlation between urea nitrogen and smoking history or triglyceride ( P>0.05). Multivariate Logistic regression analysis result showed that age, hypertension, albumin, urea nitrogen, troponin I and NT-proBNP were independent risk factors for in-hospital death in patients with heart failure ( OR = 1.018, 0.613, 0.924, 1.082, 1.340 and 1.005; 95% CI 1.002 to 1.033, 0.427 to 0.881, 0.889 to 0.961, 1.040 to 1.126, 1.111 to 1.617 and 1.003 to 1.007; P<0.05 or <0.01). ROC curve analysis result showed that the area under the curve (AUC) of urea nitrogen for prediction of in-hospital death in patients with heart failure was 0.737 (95% CI 0.728 to 0.748), and the optimal threshold value was 11.41 mmol/L, with a sensitivity of 60.16% and a specificity of 77.01%; the AUC of NT-proBNP for prediction of in-hospital death in patients with heart failure was 0.726 (95% CI 0.712 to 0.740), and there was no statistical difference in the AUC between urea nitrogen and NT-proBNP ( Z=1.055, P=0.291). Conclusions:Elevated urea nitrogen level is independently associated with an increase in in-hospital mortality in patients with heart failure, and the optimal threshold for predicting in-hospital death is 11.41 mmol/L.

Objective:To explore the relationship of telomere length, mitochondrial DNA copy number of peripheral blood with hypertension and the interaction between telomere length and mtDNA-CN on hypertension in coal miners.Methods:A case control study was conducted in a coal mine of Shanxi province from July to December of 2013, in which 325 healthy workers were selected as the control group and 378 workers with hypertension as the case group. The information about general demographic characteristics and life behavior habits of the subjects were collected through questionnaire. Levels of telomere length and mtDNA-CN in peripheral blood were detected by real-time PCR. Unconditional logistic regression was used to examine the association between hypertension and telomere length, mtDNA-CN. The interaction test between telomere length and mtDNA-CN on hypertension was performed by adding the interaction term in the corresponding model.Results:The mean telomere length of the workers in the case group was (1.50±0.55) kb, and that of the control group was (2.01±0.62) kb, the difference between two groups was significant ( t=11.68, P<0.001). The correlation analysis showed that telomere length was positively correlated with mtDNA-CN ( r=0.157, P=0.002) in the case group. Multivariate analysis showed that telomere length ( OR=4.408, 95% CI: 3.012-6.452), age ( OR=0.417, 95% CI: 0.284-0.613), BMI ( OR=1.357, 95% CI: 1.162-1.584), monthly household income level ( OR=0.656, 95% CI: 0.553-0.778) and work duration ( OR=1.249, 95% CI: 1.100-1.417) were influencing factors of hypertension. The multiply interaction between telomere length and mtDNA-CN was significant on hypertension ( OR=1.267, 95% CI: 1.094-1.468). Conclusions:The results suggest shorter telomere length is a risk factor of hypertension. There is a multiply interaction between telomere length and mtDNA-CN on hypertension. However, the association between mtDNA-CN and hypertension was not found.

[Abstract] Objective: To investigate the in vitro cytotoxicity of the chimeric antigen receptor-modified T cells and NK-92MI cells (CAR-NK-92MI cells) and CAR-CD19-T cells against mantle cell lymphoma (MCL). Methods: CAR-T cell technology, successfully obtained in clinical trial of B-lineage acute lymphoblastic leukemia (B-ALL) treatment, was used in this study. In the case of high expression of CD19 antigen in MCL, CAR- CD19-T cells and CAR- CD19-NK-92MI cells targeting CD19 antigen were generated, respectively. Then, their cytotoxicity against MCL cell lines was detected by LDH release assay and the results were verified by flow cytometry. Results: Compared with control group, both CAR-NK-92MI and CAR-CD19-T cells exhibited prominent killing effect against MCLcells（all P<0.01）; in addition, the two CAR cells exhibited high cytotoxicity against K562-CD19 cells but not on K562 cells（all P <0.01）. The death rate of MCL cells from CAR-NK-92MI group was 30%-40% more than that of control group, and the death rate of MCL from CAR-CD19-T group was 40%-50% more than that of control group. Conclusion: Both CAR-NK-92MI and CAR-CD19-T cells exhibited potent cytotoxicity against MCLcells in vitro.

Objective To compare the curative effect of conservative treatment and full thread hollow screw in the treatment of Ekrol 2 fracture of the fifth metatarsal bone.Methods From April 2013 to May 2015,42 patients with avulsion fractures of the fifth metatarsal basement of type 2 fractures in Ekrol division were treated in our hospital.23 patients underwent full thread hollow screw surgery (surgical treatment group),and there were 12 males and 11 females,with an average age of 43.4±9.2 years,and the fracture displacement length was 2.3±0.2 mm.19 patients treated with plaster fixation (conservative group),including 10 males and 9 females,with an average age of 45.7±8.9 years and fracture displacement length was 2.1±0.2 mm.During follow-up,X -ray examination was performed to evaluate the time of fracture healing and forefoot score of American Orthopaedic Foot and Ankle Society (AOFAS),visual analogue score (VAS) and quality of life questionnaire (SF-36) were compared between the two groups.Results All 42 patients were followed up for 12 to 24 months with an average of 18 months.At the latest follow-up,the fracture healing time of the surgical group was 9.3±2.0 weeks,with AOFAS score of 84.1±3.9,VAS score of 0.1±0.2,and the SF-36 score of 95.1±2.9 points,and the shortening of length after treatment was 2.1±2.5 mm;the average healing time of the conservative treatment group was 12.2±2.1 weeks,with the average AOFAS score of 84.2±4.2 points,and the average VAS score of 0.2±0.2 points,the average SF-36 score of 94.1 ±5.2 points,and the shortening of length after treatment was 0.4±2.5 mm.The results of each short-term score and the shortening of the length of the fracture after treatment was statistically significant.The results of the mid-and long-term scores showed no significant difference between the two groups;the fractures of both groups reached bone healing.3 cases of sural nerve stimulation occurred in the operation group,while 1 case occurred in the conservative group.Abnormal healing occurred in 2 cases of conservative treatment,and the fractures of the surgical treatment group were anatomic reduced.Conclusion Usingfull thread hollow screwfor the treatment of Ekrol 2 fracture of the fifth metatarsal bone is better than the conservative treatment in the short term,but there is no significant difference in the long-term effect.