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.

Objective To investigate effects of oxidized low density lipoprotein/β2 glycoprotein-Ⅰ/anti-β2 glycoprotein-Ⅰ antibody(oxLDL/β2GPⅠ/aβ2GPⅠ)complex on the proliferation,migration and angiogenesis of vascular endothelial cells and its mechanism.Methods Human umbilical vein endothelial cells(HUVEC)were cultured to logarithmic growth phase and grouped into the control group(normal culture),the oxLDL group(50 mg/L oxLDL),the oxLDL/β2GPⅠ/aβ2GPⅠ group(50 mg/L oxLDL/100 mg/L β2GPⅠ/100 mg/L aβ2GPⅠ)and the VEGF group(100 μg/L VEGF).The gene expressions of VEGF,vascular endothelial cadherin(VE-cadherin),matrix metalloproteinase(MMP)-2 and MMP-9 were detected by real-time quantitative fluorescent PCR(qPCR).Cell counting kit-8(CCK-8)method was employed to detect cell proliferation.Cell migration and invasion were determined by scratch healing test and Transwell assay.Matrigel tube formation assay was used to observe the angiogenesis of HUVEC.The relative protein expression of TLR4,MyD88 and NF-κB were examined by Western blot assay.Results Compared with the control group,the proliferation activity of cells at 48 h of treatment was increased in the oxLDL/β2GPⅠ/aβ2GPⅠ group(P＜0.05).Moreover,compared with the control group,cell migration and angiogenesis were increased in the oxLDL/β2GPⅠ/aβ2GPⅠ group,and the mRNA levels of VEGF,VE-cadherin,MMP-2 and MMP-9 were elevated(P＜0.05).Compared with the control group,levels of TLR4 and MyD88 were elevated in the oxLDL/β2GPⅠ/aβ2GPⅠ complex group(P＜0.05),as well as levels of p-NF-κB p65/NF-κB p65(P＜0.05).Conclusion oxLDL/β2GPⅠ/aβ2GPⅠ complex may promote the proliferation,migration and tube formation of vascular endothelial cells by regulating TLR4/MyD88/NF-κB signaling pathway.

Objective:To investigate the antioxidant function of Mycoplasma pneumoniae MPN662 and analyze the key active sites, and to explore the role of MPN662 in the regulation of the production of reactive oxygen species (ROS) and superoxide dismutase (SOD) in THP-1 cells. Methods:pET28a(+ )- mpn662, recombinant mutant plasmids pET28a(+ )- mpn662-Ser 66 (the 66 th Cys was mutated to Ser) and pET28a(+ )- mpn662-Ala 66 (the 66 th Cys was mutated to Ala) were constructed, recombinant proteins rMPN662, rMPN662-Ser 66 and rMPN662-Ala 66 were expressed, identified, and purified. DTNB method was employed to analyze the MetO reduction activity of rMPN662 and recombinant mutant protein. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot were applied to examine the transcription level of the mpn662 gene and the expression level of MPN662 protein after Mycoplasma pneumoniae were stimulated with different concentrations of hydrogen peroxide (H 2O 2) or tert-butyl hydroperoxide (t-BHP), respectively. Fluorescent probes (DCFH-DA) and the total SOD activity detection kit were used to test the levels of intracellular ROS and SOD in THP-1 cells, which were pretreated with rMPN662, and then stimulated by Mycoplasma pneumoniae lipid-associated membrane proteins (LAMPs). Results:Mycoplasma pneumoniae rMPN662 could reduce MetO to Met, and the enzyme activities of mutant protein were significantly lower than those of rMPN662 protein. mpn662 gene mRNA transcription level and MPN662 protein expression level were significantly increased in a dose-dependent manner when Mycoplasma pneumoniae was stimulated with H 2O 2 and t-BHP. Treatment with rMPN662 before THP-1 cells were exposed to LAMPs could decrease the level of ROS and increase the production of SOD. Conclusions:Mycoplasma pneumoniae MPN662 can reduce MetO to Met, and Cys66 is the key amino acid for this activity. MPN662 can decrease the release of ROS and increase the production of SOD in Mycoplasma pneumoniae LAMPs stimulated THP-1 cells.

Objective: To explore the most economically feasible cervical cancer screening strategies in urban China. Methods: A series of Markov models were constructed to evaluate health and economic outcomes of different screening strategies. There were 24 screening strategies including four screening methods: liquid-based cytology (LBC), human papillomavirus (HPV) DNA genotyping, HPV DNA genotyping with LBC triage (HPV DNA+ LBC), HPV DNA genotyping and LBC co-testing (HPV DNA-LBC), along with three intervals (every 1, 3 or 5 years) and two starting age for screening (30 or 35 years old) were compared. Models parameters were obtained from a cervical cancer screening study in urban China and literature reviews. Results: The cumulative incidence and mortality risk of cervical cancer declined over 69% and 82% respectively for each screening strategy as compared with the no screening scenario. LBC every five years starting from 35 years old strategy cost the least (RMB 690 per capita) and could save life years compared with no screening. The cost effectiveness ratios of 24 strategies ranged from -10 903 to 117 992 RMB per life year saved. All strategies were cost-effective compared to no screening. In the incremental cost-effectiveness analysis, LBC every 5 years starting from 30 strategy, HPV DNA genotyping every 3 years starting from 30 strategy, LBC every 3 years starting from 30 strategy and LBC every year starting from 30 strategy were dominant strategies. Conclusions Screening can effectively prevent cervical cancer. In urban Chinese areas with insufficient socioeconomic resources, LBC every 5 years from 35 years old strategy is recommended. In relatively more affluent areas, LBC every 5 years from 30 years old strategy, LBC every 3 years from 30 years old strategy, HPV DNA genotyping every 3 years from 30 years old strategy, and LBC every year from 30 years old strategy are recommended successively.

Objective: To investigate the effects of oxidized low-density lipoprotein/β2 glycoproteinⅠ/β2 glycoproteinⅠantibody (oxLDL/β2GPⅠ/β2GPⅠ-Ab) complex on the phenotypic transformation and lipid transpoters on the surface of rat thoracic aorta smooth muscle cell line (A7r5), and their correlation with toll-like receptor 4 (TLR4) signaling pathway. Methods: A7r5 cells were stimulated by oxLDL, oxLDL/β2GPⅠ complex, oxLDL/β2GPⅠ-Ab complex, β2GPⅠ/β2GPⅠ-Ab complex and oxLDL/β2GPⅠ/β2GPⅠ-Ab complex respectively, and then total RNA and protein were collected. The expressions of α-smooth muscle actin (α-SMA), macrophage surface marker CD68, galectin-3 (LGALS3), scavenger receptor class B member 3 (CD36) and ATP-binding cassette transporter A1/G1 (ABCA1/ABCG1) were detected by real-time quantitative PCR (RT-qPCR), western blot and immunofluorescence (IF) respectively. The roles of TLR4 and its downstream signaling molecules in the phenotypic transformation and expression changes of lipid transporters of A7r5 cells induced by oxLDL/β2GPⅠ/β2GPⅠ-Ab complex were investigated by the pretreatment of TLR4 blocker TAK-242 (5 μmol/L) or c-Jun N-terminal kinases 1/2 (JNK 1/2) blocker SP600125 (90 nmol/L). Results: The oxLDL/β2GPⅠ/β2GPⅠ-Ab complex significantly increased the levels of CD68 and LGALS3, and decreased the level of α-SMA, while TAK-242 could reverse this phenomenon. The oxLDL/β2GPⅠ/β2GPⅠ-Ab complex could promote the expression of CD36 and inhibit the expression of ABCA1/ABCG1, while TAK-242 and SP600125 could reverse this process. Conclusion The oxLDL/β2GPⅠ/β2GPⅠ-Ab complex promotes the phenotypic transformation of A7r5 cells to macrophage-like cells, regulates the expression of lipid transport-related molecules and enhances the ability of lipids transport into cells. TLR4 and JNK1/2 are closely related to this process.

Objective: To investigate the effects of β2 glycoprotein Ⅰ/anti-β2 glycoprotein Ⅰ complex (β2/aβ2) on oxidized low density lipoprotein (oxLDL)-mediated lipid accumulation and focal adhesion kinase (FAK) activation in THP-1 macrophage, as well as the role of Toll-like receptor 4 (TLR4) during the process. Methods: THP-1 cells were differentiated into THP-1 macrophage by PMA (100 ng/mL). THP-1 macrophages were treated with RPMI 1640 medium, oxLDL, oxLDL+β2/aβ2 or oxLDL+lipopolysaccharide (LPS). The mRNA expressions of lipid transportation molecules, ACAT1, ABCA1 and ABCG1 were detected by RT-qPCR. Intracellular total cholesterol (TC) and free cholesterol (FC) in THP-1 macrophages were evaluated by Trinder assay, then the content and proportion of intracellular cholesteryl ester (CE) were calculated. The expression and phosphorylation of FAK were detected by immune fluorescence, RT-qPCR and western blot. To evaluate the role of TLR4, THP-1 macrophages were pre-treated with or without TLR4 inhibitor TAK-242 (1 μg/mL). Results: β2/aβ2 treatment significantly inhibited oxLDL-mediated lipid accumulation and FAK expression and phosphorylation in THP-1 macrophages, which could be reversed by TLR4 blockage. Conclusion β2/aβ2 inhibits the oxLDL-mediated lipid accumulation and FAK activation of THP-1 macrophage, which is related to the function of TLR4.

Objective: To account the direct cost of uterine cervix carcinoma treatment in China and to explore the related factors which influence the direct financial burden of the disease. Methods: Data was collected through the medical record system and telephone interviews in 14 county-level hospitals and 9 provincial and municipal hospitals from 14 provinces/municipalities enrolled in the Chinese National Health Industry Research Project in 2015. The direct financial burden of uterine cervix carcinoma treatment consisted of the direct medical cost and the direct non-medical cost of treatment in different pathological cervical cancer stages and precancerous lesions. Multiple liner regression method was used to analyze the factors affecting the costs. Results: The age of the 3 246 patients was (46.40±10.43) years, including 2 423 patients from provincial and municipal hospitals and 823 patients from county-level hospitals. The direct financial burden for one patient of pathological uterine cervix carcinoma stage or precancerous lesion ranged from 10 156.3 yuan to 75 716.4 yuan in provincial and municipal hospitals, and for patients from county-level hospitals, the cost was between 4 927.9 yuan and 47 524.8 yuan per person. There was a wide gap between the direct financial burden of patients in different disease stages. The direct financial burden of patients with precancerous lesions ranged from 4 927.9 yuan per person to 11 243.0 yuan per person, as for patients of pathological uterine cervix carcinoma stages, the direct financial burden was between 29 274.6 yuan and 75 716.4 yuan per person. The factors which influence direct financial burden would include: the levels of the hospital, pathological period, medicare reimbursement, days of treatment, and the methods of treatment (P<0.001). Conclusion The direct financial burden of diseases in patients with pathological uterine cervix carcinoma stage or precancerous lesion differed in different levels of hospital and pathological periods. In addition, medicare reimbursement, days of treatment, and the methods of treatment all had impact on it.

Objective To compare the short-term clinical effects of totally thoracoscopic surgery and traditional open-heart sugery in the treatment of esophageal cancer. Methods 64 patients with middle thoracic esophageal cancer admitted in our hospital from June 2013 to June 2016 were selected. Among them, 32 patients underwent totally thoracoscopic surgery were identified as totally thoracoscopic group and the remaining 32 patients underwent traditional open-heart surgery were identified as control group. We compared various surgical indicators of patients in the two groups. Results In the totally thoracoscopic group, the duration of operation was(123. 8±25. 1) min, the amount of blood loss during surgery was (172. 1±30. 3) mL, the retention time of chest drainage tube was (3. 1±1. 1) d and the duration of hospitalization was (15. 6±2. 7) d. Compared with the control group, these indicators showed significant difference(P＜0. 05). The incidence of postoperative complications in the totally thoracoscopic group was 9. 3％, which was significantly lower than that in the control group(31. 3％)(P＜0. 05). The pain severity of totally thoracoscopic group reduced significantly than the control group(P＜0. 05). Conclusion Compared with the traditional open-heart surgery, totally thoracoscopic surgery for esophageal cancer has advantages of less bleeding, less trauma, few complications and less pain, which is worthy of promoting and using widely in clinic.

OBJECTIVETo investigate peripheral blood apelin levels in patients with acute ST-elevation myocardial infarction (STEMI) and their correlation with the one-year outcome of the patients.

METHODSA total of 153 consecutive patients, including 93 with acute STEMI undergoing primary percutaneous coronary intervention (PCI), 30 with acute STEMI and 30 with stable angina all undergoing elective PCI, and 10 healthy control subjects were examined for peripheral blood apelin levels and clinical parameters. The composite endpoints (CEPs) were determined at the one year follow-up.

RESULTSApelin levels were significantly decreased in all the patients at admission, but increased following primary PCI. Apelin levels showed a negative correlation with glycosylated hemoglobin levels. At one year following PCI, the patients with a lower apelin level showed an increased risk for lowered left ventricular ejection fraction ratio, but further analysis failed to provide evidence that apelin levels were predictive of the one-year outcome.

CONCLUSIONPeripheral blood apelin levels might be useful for predicting the clinical outcomes of patients with acute STEMI.

Acute Disease ; Apelin ; Biomarkers ; blood ; Case-Control Studies ; Humans ; Intercellular Signaling Peptides and Proteins ; blood ; Myocardial Infarction ; blood ; diagnosis ; Percutaneous Coronary Intervention ; Prognosis ; Ventricular Function, Left

Objective To investigate peripheral blood apelin levels in patients with acute ST-elevation myocardial infarction (STEMI) and their correlation with the one-year outcome of the patients. Methods A total of 153 consecutive patients, including 93 with acute STEMI undergoing primary percutaneous coronary intervention (PCI), 30 with acute STEMI and 30 with stable angina all undergoing elective PCI, and 10 healthy control subjects were examined for peripheral blood apelin levels and clinical parameters. The composite endpoints (CEPs) were determined at the one year follow-up. Results Apelin levels were significantly decreased in all the patients at admission, but increased following primary PCI. Apelin levels showed a negative correlation with glycosylated hemoglobin levels. At one year following PCI, the patients with a lower apelin level showed an increased risk for lowered left ventricular ejection fraction ratio, but further analysis failed to provide evidence that apelin levels were predictive of the one-year outcome. Conclusion Peripheral blood apelin levels might be useful for predicting the clinical outcomes of patients with acute STEMI.