Lipid metabolism, BMI and the risk of nonalcoholic fatty liver disease in the general population: evidence from a mediation analysis.

BACKGROUND: Body mass index (BMI) and lipid parameters are the most commonly used anthropometric parameters and biomarkers for assessing nonalcoholic fatty liver disease (NAFLD) risk. This study aimed to assess and quantify the mediating role of traditional and non-traditional lipid parameters on the association between BMI and NAFLD. METHOD: Using data from 14,251 subjects from the NAGALA (NAfld in the Gifu Area, Longitudinal Analysis) study, mediation analyses were performed to explore the roles of traditional [total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C)] and non-traditional [non-HDL-C, remnant cholesterol (RC), TC/HDL-C ratio, LDL-C/HDL-C ratio, TG/HDL-C ratio, non-HDL-C/HDL-C ratio, and RC/HDL-C ratio] lipid parameters in the association of BMI with NAFLD and quantify the mediation effect of these lipid parameters on the association of BMI with NAFLD using the percentage of mediation. RESULT: After fully adjusting for confounders, multivariate regression analysis showed that both BMI and lipid parameters were associated with NAFLD (All P-value < 0.001). Mediation analysis showed that both traditional and non-traditional lipid parameters mediated the association between BMI and NAFLD (All P-value of proportion mediate < 0.001), among which non-traditional lipid parameters such as RC, RC/HDL-C ratio, non-HDL-C/HDL-C ratio, and TC/HDL-C ratio accounted for a relatively large proportion, 11.4%, 10.8%, 10.2%, and 10.2%, respectively. Further stratified analysis according to sex, age, and BMI showed that this mediation effect only existed in normal-weight (18.5 kg/m2 ≤ BMI < 25 kg/m2) people and young and middle-aged (30-59 years old) people; moreover, the mediation effects of all lipid parameters except TC accounted for a higher proportion in women than in men. CONCLUSION: The new findings of this study showed that all lipid parameters were involved in and mediated the risk of BMI-related NAFLD, and the contribution of non-traditional lipid parameters to the mediation effect of this association was higher than that of traditional lipid parameters, especially RC, RC/HDL-C ratio, non-HDL-C/HDL-C ratio, and TC/HDL-C ratio. Based on these results, we suggest that we should focus on monitoring non-traditional lipid parameters, especially RC and RC/HDL-C ratio, when BMI intervention is needed in the process of preventing or treating NAFLD.

rs35301225 polymorphism in miR-34a promotes development of human colon cancer by deregulation of 3'UTR in E2F1 in Chinese population.

BACKGROUND: Previous reports have revealed that down-regulation of miR-34a expression can promote colorectal cancer (CRC) cell growth by targeting cell cycle-related transcriptional factor E2F1. To date, the function of the single nucleotide polymorphism (SNP) located in the mature region of miR-34a has not been investigated. METHODS: We performed a case-control study including 685 CRC patients and 618 cancer-free controls. Genotyping, real-time PCR assay, cell transfection, and the dual luciferase reporter assay were used in our study. Cell proliferation and cell cycle analysis were measured in CRC cells including Hct-116 and SW480. The overall survival of different genotypes was also investigated. RESULTS: We found that the rs35301225 polymorphism in miR-34a was involved in the occurrence of CRC by acting as a tumor suppressor by down-regulation of tumor-promoting gene E2F1. C/A SNP of miR-34a could promote CRC cell proliferation by up-regulation of E2F1. Also, C/A genotype can change the cell cycle by increasing the S phase percentage. Moreover, the SNP in rs35301225 of miR-34a was associated with tumor size and tumor differentiation, as well as metastasis in CRC patients; C/A SNP was related to the significantly enhanced expression of E2F1 and shorter survival in post-surgery CRC patients. CONCLUSIONS: rs35301225 in miR-34a was highly associated with a decreased risk of CRC in a Chinese population and might serve as a novel biomarker for colon cancer.

Association of mean arterial pressure with non-alcoholic fatty liver disease: results from the NAGALA study.

Objective: Several recent reports have suggested the use of mean arterial blood pressure (MAP) to assess/predict the risk of developing atherosclerosis, chronic kidney disease, diabetes, metabolic syndrome, and poor prognosis in a variety of cardiovascular and cerebrovascular diseases. The current study aimed to investigate the association of MAP with non-alcoholic fatty liver disease (NAFLD) and to explore the differences in this association across populations. Methods: This study used data from the NAGALA study from 1994 to 2016. MAP was calculated as 1/3 systolic blood pressure (SBP) + 2/3 diastolic blood pressure (DBP). Restricted cubic spline (RCS) and logistic regression models were used to examine the correlation of MAP with NAFLD. Results: The study population was 14,251 general people undergoing health screening, with a median (interquartile range) age of 42 (36-50) years; among them, 48% were women, and 2,507 (17.59%) were diagnosed with NAFLD. After fully controlling for confounders in the current dataset, MAP was positively and non-linearly associated with NAFLD [(odds ratios (ORs): 1.39, 95% confidence intervals (CIs): 1.15, 1.68); P for non-linearity = 0.024]; the dose-response curve showed that there was a transient saturation effect interval when MAP was between 85 and 95 mmHg, where the risk of NAFLD was neither increased nor decreased. The results of the stratified analysis showed that the risk of NAFLD associated with MAP appeared to be influenced only by age (P-interaction = 0.002), but not by sex, body mass index (BMI), habits of exercise, drinking status, or smoking status (P-interaction > 0.05); further age-stratified RCS analysis showed that the non-linear association between MAP and NAFLD in the young and middle-aged and the middle-aged and elderly populations was consistent with the results of the whole population, whereas, in the elderly population, a U-shaped curve association between MAP and NAFLD was observed, with both low and high MAP increasing the risk of NAFLD. Conclusion: In the general population, MAP was positively and non-linearly associated with NAFLD, and this association only differed significantly by age, but not by sex, BMI, habits of exercise, drinking status, and smoking status.

Phenotypic and genotypic characterization of multi-drug resistance Pseudomonas aeruginosa isolated from urinary tract infections of non-catheterized and catheterized Chinese patients: A descriptive study over 3 years.

Urinary tract infections (UTI) are commonest infections, especially in catheterized patients. It is responsible of mortality and morbidity among hospitalized patients. The objectives of the study were to demonstrate the virulence factors and their genes of multi-drug resistance Pseudomonas aeruginosa causing UTI. A total of 366 non-catheterized and 171 catheterized patients' (in whom the catheter was in > 48 hours duration) urine samples (one sample/patient) from both sexes were collected and processed. >105 colony forming unit was considered as Pseudomonas aeruginosa culture-positive. Antimicrobial susceptibility testing was done by the Kirby Bauer disc diffusion method (The Clinical and laboratory standards institute guidelines 2019). The virulence factors were detected by in vitro assay method and polymerase chain reaction was done to detect the resistance genes present in Pseudomonas aeruginosa. Biofilm production was detected by the microtiter plate method. Out of 537 urine samples a total of 280 (52%) were females and 257 (48%) were male patients. Out of 366 non-catheterized urine samples 42 (23.6%) grew Pseudomonas aeruginosa and out of 171 catheterized urine 23 (25.84%) grew Pseudomonas aeruginosa. All were multi-drug resistance strains. A total of 10 (23.80%), 42 (100%), 8 (19.05%), 24 (57.14%), and 36 (85.71%) produced the Metallo-ß-lactamases, AmpC-ß-lactamase, carbapenemase, strong biofilm, and twitching motility positive, respectively in non-catheterized urine samples. A total of 11, 34, 9, 28, and 37 were oxacillinases-23, multidrug efflux protein resistance, New Delhi metallo-ß-lactamase-1, Verona Integron-encoded MBL, and Pseudomonas specific enzyme gene detected in non-catheterized urine samples. A total of 8 (34.8%), 6 (26.01%), 4 (17.39%), 15 (65.2%), and 18 (78.26%) were produced Metallo-ß-lactamases, carbapenemase, AmpC-ß-lactamase, strong biofilm, and twitching motility positive, respectively in catheterized urine samples. A total of 6, 18, 4, 16, and 15 were oxacillinases 23, multidrug efflux protein resistance, New Delhi metallo-ß-lactamase-1, Verona Integron-encoded MBL, and Pseudomonas specific enzyme, respectively genes detected in catheterized urine samples. Biofilm formation and twitching motility showed correlation among culture-positive Pseudomonas aeruginosa strains from catheterized patients (Correlation coefficients = 6.2, 95% confidence interval: 5.4-7.2). A better hospital infection control practice and detailed investigation of the microevolution of Pseudomonas aeruginosa in UTI are needed.

Clinical Value of Pepsinogen in the Screening, Prevention, and Diagnosis of Gastric Cancer.

OBJECTIVES: To compare the levels of serum pepsinogen (PG) in patients with gastric cancer (GC), patients with atrophic gastritis (AG), and healthy donors. Also, we explored the clinical value of PG detection for the diagnosis and treatment of GC. METHODS: The PG level in peripheral blood from patients and heathy donors was determined using an Abbott automatic chemiluminescence instrument. The study included 117 patients with GC confirmed by gastroscopy and histopathology, of whom 13 patients had cancer at stage I, 47 at stage II, 41 at stage III, and 16 at stage IV. The AG group included 122 patients, and the control group had 120 healthy donors. The relationship between serum PG levels and the occurrence and development of GC, as well as the evaluation of the clinical value of diagnostic tests based on serum PG detection, were investigated by receiver operating characteristic (ROC) curve analyses. RESULTS: Pepsinogen I (PGI) levels gradually decreased from the control group, the AG group, and the GC group. PGI exhibited high diagnostic value for GC (area under the curve [AUC], 0.834; cutoff, 51.2 ng/mL, sensitivity, 81.7%; specificity, 68.4%), PGII (AUC, 0.587; cutoff value, 13.05 ng/mL; sensitivity, 65.8%; specificity, 53.8%), and PGR (AUC, 0.752; cutoff, 5.65; sensitivity, 54.2%; specificity, 87.2%). The occurrence of GC was negatively correlated with serum levels of PGI (B = -0.054; OR = 0.947; 95% confidence interval [CI], 0.925-0.970; P <.001) and PGR (B = -0.420; OR = 0.657; 95% CI, 0.499-0.864; P = .003). CONCLUSIONS: The combined detection of PGI, PGII, and PGR has important clinical value for the screening, prevention, and diagnosis of GC and could allow for earlier detection, diagnosis, and treatment of GC.

A Novel Combined DNA Vaccine Encoding Toxoplasma gondii SAG1 and ROP18 Provokes Protective Immunity Against a Lethal Challenge in Mice.

PURPOSE: Antigens expressed by Toxoplasma gondii (T. gondii) during its life cycle trigger various immune responses in the host. Recently, toxoplasma vaccine research focused on T. gondii surface antigen 1 (SAG1) and Rhoptry Protein 18 (ROP18) to establish a safe and efficacious DNA vaccine. METHOD: We constructed two eukaryotic expression plasmids: p3 × FLAG-Myc-CMV™-24-SAG1 and p3 × FLAG-Myc-CMV™-24-ROP18. BALB/c mice were randomly divided into six groups and immunized with these DNA vaccines either separately or in combination. The combination vaccine was administered at either the full dose or at half-strength dose. Control mice were immunized with empty vector or with phosphate-buffered saline. RESULTS: The frequency of CD4+ cells in the spleen was consistent among all groups, whereas that of CD8+ T cells was the highest in the group immunized with the combination vaccine at half-strength dose (p < 0.05). Importantly, the mRNA expression levels of interleukin-12 (IL-12) and interferon-gamma (INF-γ) were closely correlated (r = 0.6, p < 0.0001) and both were upregulated in the group that was immunized with the combination vaccine at half-strength dose (p < 0.0001). The survival time of the mice subjected to a lethal dose of toxoplasma was significantly extended by prior immunization with DNA vaccines expressing either SAG1 or ROP18 or a combination of both (p < 0.05). The group that was immunized with the combination vaccine at half-strength dose demonstrated the best efficacy (p < 0.05). CONCLUSION: These results showed that the combination DNA vaccine provided better immune protection than the single gene vaccines, and that optimizing the dosing of the vaccine can improve the immune response.

Synergistically increased ILC2 and Th9 cells in lung tissue jointly promote the pathological process of asthma in mice.

In recent years, T helper (Th) 9 cells have been demonstrated to be key mediators in immune responses in asthmatic lungs, and innate lymphoid cells 2 (ILC2s) have been described as a novel type of innate immunocyte with the ability to enhance immunoglobulin E (IgE) production. However, the interaction between ILC2s and Th9 cells in the pulmonary system of a mouse model of asthma remains to be elucidated. In the present study, the response state of lung tissue with regards to Th9 and ILC2s in a mouse model of asthma was investigated by detecting Th9­ and ILC2­associated cytokine receptors. The present study also investigated the association between the expression levels of the cytokine receptors in lung tissue samples and the IgE levels in sera samples from mouse models of asthma. Results from the present study demonstrated that the frequency of ILC2s and Th9 cells was significantly increased in the lung tissue samples, indicating that a Th2-type immune response had occurred. In addition, high mRNA expression levels of RAR­related orphan receptor α, interleukin 1 receptor­like 1, transcription factor PU.1 and interleukin (IL)­9 were observed. Furthermore, IL­5Rα, IL­13Rα2 and high­affinity IgE receptor were increased in mouse models of asthma, and a positive association was observed between the expression levels of ILC2­ or Th9­associated receptors in tissue samples and IgE levels in the sera. This indicated that ILC2s and Th9 were in a state of polarization and may promote each other in the lung tissue of mouse models of asthma, and that the lung tissue was responding to the two types of cells via increased expression of receptors.

Arg97² insulin receptor substrate-1 inhibits endothelial nitric oxide synthase expression in human endothelial cells by upregulating microRNA-155.

The dysregulation of nitric oxide (NO) synthesis attributable to the abnormal expression/activity of endothelial NO synthase (eNOS) is considered to be a major characteristic of insulin-resistant states, as well as an essential contributor to the pathogenesis of cardiovascular diseases. The Arg972 insulin receptor substrate-1 (IRS-1) is associated with insulin resistance. In the present study, we investigated the association between Arg972 IRS-1 and eNOS expression/activity in human subjects and in primary cultures of human endothelial cells. Data from 832 human subjects revealed that heterozygous and homozygous Arg972 IRS-1 carriers had significantly lower levels of plasma eNOS and nitrite/nitrate than the homozygous wild-type (WT) IRS-1 carriers. Human umbilical vein endothelial cells (HUVECs) established from delivering mothers expressing heterozygous Arg972 IRS-1 had significantly lower eNOS expression/activity and higher miR-155 levels than those expressing WT homozygous IRS-1. The overexpression of IRS-1 and Arg972 IRS-1 in the HUVECs, respectively, decreased and increased the miR-155 expression level. In addition, the overexpression of IRS-1 in the HUVECs significantly increased eNOS expression; this effect was reversed by transfection with mature miR-155 mimic or treatment with the selective phosphatidylinositol-3 kinase (PI3K) inhibitor, BKM120. On the other hand, the overexpression of Arg972 IRS-1 markedly decreased eNOS expression and this effect was reversed by transfection with antagomir-155. On the whole, our in vivo data demonstrate that Arg972 IRS-1 is associated with decreased plasma eNOS and nitrite/nitrate levels in human subjects. Our in vitro data demonstrate that Arg972 IRS-1 inhibits eNOS expression in human endothelial cells by upregulating miR-155 expression through the impairment of PI3K signaling. The present study provides new insight into the pathophysiological role of Arg972 IRS-1 in cardiovascular diseases.

Characterization and distribution of drug resistance associated ß-lactamase, membrane porin and efflux pump genes in MDR A. baumannii isolated from Zhenjiang, China.

BACKGROUND: Acinetobacter baumannii (A. baumannii), especially the multidrug resistant A. baumannii (MDR-AB) is becoming a common opportunistic pathogen in hospital, and constitutes significant public health threats. This study aimed at investigating the relationship between drug resistance with expression of class A-D ß-lactamase genes, mutation in membrane porin and over-expression of efflux pump genes among A. baumannii isolated from Zhengjiang, China. METHODS: Antibiotic susceptibility assays were performed using Kirby-Bauer disc diffusion method. PCR was used to detect ß-lactamase genes and carO, oprD, adeR, adeS. Real-time PCR was used to assess the mRNA expression level of efflux pump gene adeB. The software of DNAMAN was applied to assemble oprD and carO sequences, and the sequences were compared with those retrieved from GenBank (http://www.ncbi.nlm.nih.gov/). RESULTS: 27 isolates (61.4%) in this study were MDR-AB, in which five ß-lactamases including TEM, CTX-M-2, ADC, OXA-23 and OXA-51 were found, and the positive rate was 96.3% (26), 14.8% (4), 92.6% (25), 88.9% (24) and 92.6% (25), respectively. In addition, the expression level of adeB mRNA was significantly increased in MDR-AB, it might due to adeR mutation. Some mutations were also found in carO and oprD. CONCLUSION: MDR-AB showed high relationship with ß-lactamase, mutation in membrane porin and overexpression of adeB, which may directly relates to the mutation in regulating gene adeR.

Solamargine triggers hepatoma cell death through apoptosis.

Solamargine (SM), a steroidal alkaloid glycoside extracted from the traditional Chinese herb Solanum incanum, has been evidenced to inhibit the growth and induce apoptosis in a number of human cancer cell lines. In the present study, the anticancer effect of SM and underlying molecular mechanism of SM-induced apoptosis were investigated on the human hepatocellular carcinoma cells, SMMC7721 and HepG2. The proliferation effects of SM on the SMMC7721 and HepG2 cell lines were evaluated using MTT and colony formation assays. In addition, the percentage of apoptosis was measured using an Annexin V/propidium iodide staining method and the cell cycle distribution mediated by SM was analyzed using flow cytometry. The expression levels of B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), caspase-3, caspase-9, proliferating cell nuclear antigen (pcna) and Ki67 proteins were examined to further demonstrate the proliferate and apoptosis effects of SM on the hepatoma cells. The results indicated that SM effectively inhibited hepatoma cell proliferation and promoted apoptosis. SM resulted in cell cycle arrest at the G2/M phase in the two cell lines. In addition, SM downregulated the levels of proliferation-associated (Ki67 and pcna) and anti-apoptotic (Bcl-2) proteins, and promoted the activity of apoptosis-associated proteins (Bax, caspase-3 and caspase-9). Therefore, the activation of the Bcl-2/Bax and caspase signaling pathways may be involved in the SM-induced apoptosis of hepatoma cells.

Increased frequencies of nuocytes in peripheral blood from patients with Graves' hyperthyroidism.

Newly identified nuocytes play an important role in Th2 cell mediated immunity such as protective immune responses to helminth parasites, allergic asthma and chronic rhinosinusitis. However, the contributions of nuocytes in the occurrence and development of Graves' hyperthyroidism remains unknown. Previous studies found that there was a predominant Th2 phenotype in patients with Graves' hyperthyroidism, it might relate to polarization of nuocytes. Nuocytes were defined by transcription factor RORα, various cell surface markers (T1/ST2, IL-17RB, ICOS, CD45) and associated cytokines. In this study, these cells related genes or molecules in PBMC from patients with Graves' hyperthyroidism were measured, and the potential correlation between them was analyzed. The expression levels of T1/ST2, IL-17RB, ICOS, IL-5 and IL-13, which represented nuocytes associated molecules were significantly increased in patients, meanwhile, the RORα mRNA also had a tendency to increase. In addition, IFN-γ and T-bet (Th1 related cytokine and transcription factor) were obviously decreased, and there was a positive correlation between IL-17RB and IL-13. These results suggested that there were polarized nuocytes in Graves' hyperthyroidism patients, and which closely related to the down-regulation of Th1 cells or relatively advantage of Th2 differentiation.

Polarization of ILC2s in peripheral blood might contribute to immunosuppressive microenvironment in patients with gastric cancer.

Newly identified nuocytes or group 2 innate lymphoid cells (ILC2s) play an important role in Th2 cell mediated immunity such as protective immune responses to helminth parasites, allergic asthma, and chronic rhinosinusitis. However, the contributions of ILC2s in the occurrence and development of cancer remain unknown. Our previous study found that there was a predominant Th2 phenotype in patients with gastric cancer. In this study, the ILC2s related genes or molecules in PBMC from patients with gastric cancer were measured, and the potential correlation between them was analyzed. The expression levels of RORα, GATA3, T1/ST2, IL-17RB, CRTH2, IL-33, IL-5, and IL-4 mRNA were significantly increased in patients, but no significant changes were found in ICOS, CD45, and IL-13 expression, and there was a positive correlation between RORα or IL-13 and other related factors, such as ICOS and CD45. The increased frequency of ILC2s was also found in PBMC of patients by flow cytometry. In addition, the mRNA of Arg1 and iNOS were also significantly increased in patients. These results suggested that there are polarized ILC2s in gastric cancer patients which might contribute to immunosuppressive microenvironment and closely related to the upregulation of MDSCs and M2 macrophages.

[Relationship between chronic congestive heart failure and sleep-disordered breathing in elderly patients].

OBJECTIVE: To determine the prevalence of sleep disordered breathing (SDB) in elderly patients with chronic congestive heart failure (CHF) and explore the relations between SDB and left ventricular function. METHODS: By means of polysomnography, 56 elderly patients with CHF were divided into non-SDB, mild SDB, moderate SDB, and severe SDB groups, and the left ventricular ejection fraction (LVEF) was measure by (99)Tc equilibrium radionuclide angiography. RESULTS: In the 56 elderly patients with CHF, 38 (67.9%) had SDB, including 12 (21.4%) mild SDB, 14 (25.0%) moderate SDB, and 12 (21.4%) severe SDB patients. Thirty (53.6%) of the 56 patients with CHF had obstructive sleep apnea (OSA), 4 (7.1%) had central sleep apnea and 22 (39.2%) had mixed sleep apnea. The moderate and severe SDB groups had lower minimum arterial oxyhemoglobin saturation during sleep than the non-SDB groups, and the apnea-hyponea index was closely related to LVEF (r=-0.74, P<0.01). CONCLUSION: The prevalence of SDB, predominantly OSA, is high in elderly patients with CHF. Moderate and severe SDB might affect the left ventricular function in these patients, who require polysomnography monitoring.