The Effect of Salvianolic Acid A on Tumor-Associated Macrophage Polarization and Its Mechanisms in the Tumor Microenvironment of Triple-Negative Breast Cancer.

Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer, with a high degree of malignancy and poor prognosis. Tumor-associated macrophages (TAMs) have been identified as significant contributors to the growth and metastasis of TNBC through the secretion of various growth factors and chemokines. Salvianolic acid A (SAA) has been shown to have anti-cancer activities. However, the potential activity of SAA on re-polarized TAMs remains unclear. As there is a correlation between the TAMs and TNBC, this study investigates the effect of SAA on TAMs in the TNBC microenvironment. For that purpose, M2 TAM polarization was induced by two kinds of TNBC-conditioned medium (TNBC-TCM) in the absence or presence of SAA. The gene and protein expression of TAM markers were analyzed by qPCR, FCM, IF, ELISA, and Western blot. The protein expression levels of ERK and p-ERK in M2-like TAMs were analyzed by Western blot. The migration and invasion properties of M2-like TAMs were analyzed by Transwell assays. Here, we demonstrated that SAA increased the expression levels of CD86, IL-1ß, and iNOS in M2-like TAMs and, conversely, decreased the expression levels of Arg-1 and CD206. Moreover, SAA inhibited the migration and invasion properties of M2-like TAMs effectively and decreased the protein expression of TGF-ß1 and p-ERK in a concentration-dependent manner, as well as TGF-ß1 gene expression and secretion. Our current findings for the first time demonstrated that SAA inhibits macrophage polarization to M2-like TAMs by inhibiting the ERK pathway and promotes M2-like TAM re-polarization to the M1 TAMs, which may exert its anti-tumor effect by regulating M1/M2 TAM polarization. These findings highlight SAA as a potential regulator of M2 TAMs and the possibility of utilizing SAA to reprogram M2 TAMs offers promising insights for the clinical management of TNBC.

Association of Red Meat Usual Intake with Serum Ferritin and the Risk of Metabolic Syndrome in Chinese Adults: A Longitudinal Study from the China Health and Nutrition Survey.

OBJECTIVE: The present study aimed to investigate the association of red meat usual intake with metabolic syndrome (MetS), and explore the contribution of red meat usual intake to serum ferritin. METHODS: Based on the data from the longitudinal China Health and Nutrition Survey (CHNS), 2,797 healthy adults aged 18-75 years without hypertension, diabetes, and MetS were selected in 2009 as subjects and follow-up studies were carried out till 2015. We used the National Cancer Institute (NCI) method to estimate the usual intake of foods. Multivariable logistic regressions were performed to evaluate the association between red meat usual intake and the risk of MetS. Quantile regression analysis was used to study the relationship between red meat consumption and serum ferritin levels. RESULTS: After adjusting for potential confounders, red meat, and fresh red meat were positively associated with the risk of MetS ( RR = 1.41, 95% CI: 1.05-1.90 and RR = 1.37, 95% CI: 1.02-1.85, respectively). These relationships showed increasing trend ( P < 0.05). The level of serum ferritin increased significantly with the number of MetS components ( P < 0.05). The quantile regression analysis showed that red meat and fresh red meat usual intake had a significant positive association with serum ferritin levels across the entire conditional serum ferritin distribution ( P < 0.05). Processed red meat did not exhibit a similar association. CONCLUSION: Higher red meat usual intake was associated with an increased risk of MetS and elevated serum ferritin levels.

Epidemics of overweight and obesity among growing childhood in China between 1997 and 2009: Impact of Family Income, Dietary Intake, and Physical Activity Dynamics.

BACKGROUND: Obesity has become a major health problem among children and adolescents worldwide. This study aimed to examine the trends of overweight and obesity among childhood in China and assess their associations with family income, dietary intake, and physical activity (PA) between 1997 and 2009. METHODS: Two waves of cross-sectional data of Chinese children and adolescents aged 7-17 years from the China Health and Nutrition Survey were used. Weight and height were measured following standardized procedures. Dietary intake was assessed by 3 consecutive 24-h recalls. Childhood overweight and obesity were defined using the International Obesity Task Force-recommended body mass index cut-offs. Multivariate linear regression analysis was used to examine the associations of family income with diet intakes and PA. Multivariate logistic regression analysis was conducted to assess the associations of overweight and obesity with family income, dietary intake, and PA. RESULTS: The prevalence of childhood overweight and obesity increased from 12.6% in 1997 to 22.1% in 2009, particularly in the medium- and high-family income groups, which increased by 102.7% and 90.3%, respectively. Higher fat intake (% energy), and moderate and vigorous PA were significantly associated with overweight and obesity in final model (odds ratio [OR] = 1.01, 95% confidence interval [CI]: 1.00-1.02, P = 0.004; and OR = 0.99, 95% CI: 0.98-1.00, P = 0.036, respectively). CONCLUSIONS: The prevalence of overweight and obesity among Chinese children and adolescents has increased between 1997 and 2009. Reducing fat intake and increasing PA may help obesity prevention.

[Proteomic study on effect of tangcao pill on microsome CYP450].

Tangcao pill is commonly applied in adjuvant and even alternative therapy for patients with AIDS. However, the herb contains complex ingredients, but with unknown effect against anti-HIV drug and unknown function. Because CYP450 emzyme is the main metabolic enzymes of the drug, it is of important significance to study the regulation of CYP450 enzymes before and after the combined administration of Tangcao pill and EFV. Proteomics, due to its high throughout and high sensitivity, has been widely applied in CYP450 enzyme study. In this paper, liver microsomes were separated through differential centrifugation. Their proteins were separated through SDS-PAGE. The three protein bands that CYP450 enzymes were located were cut and identified by liquid chromatography tandem mass spectrometry. Totally 16 CYP450 isoenzymes were identified. Furthermore, in order to make a quantitative analysis on the effect of tang herb on CYP450 emzyme, the multiple reaction monitoring (MRM) technology based on MS was adopted. The CYP2C11 was selected based on the results of the mass spectrum identification of proteins. The characteristic polypeptides were obtained through searching Expasy blast database. The m/z of the fragment ions was less than 800. In the paper, the m/z of ion pairs of CYP2C11 were 711.5/232.1, 711.5/319.2, 711.5/466.2 and 711.5/595.3, and the m/z of ESAT-6 (internal standard, IS) were 735.5/215.3, 735.5/389.3, 735.5/460.3 and 735.5/524.3. The relative peak (analyte/IS) area was adopted for the relative quantitative analysis. Compared with the EFV single administration group, the EFV and Tangcao pill combined administration group showed a 1.6-fold increase in CYP2C11. The results of the paper indicated that Tangcao pill may affect drug metabolism by regulating metabolic enzymes such as CYP2C11, but the specific mechanism still unknown.

Serum dihydroxyacetone kinase peptide m/z 520.3 as predictor of disease severity in patients with compensated chronic hepatitis B.

BACKGROUND AND AIM: Due to known limitations of liver biopsy, reliable non-invasive serum biomarkers for chronic liver diseases are needed. We performed serum peptidomics for such investigation in compensated chronic hepatitis B (CHB) patients. METHODS: Liquid chromatography combined with tandem mass spectrometry (LC-MS/MS) was used to identify differentially expressed peptides in sera from 40 CHB patients (20 with S0G0-S1G1 and 20 with S3G3-S4G4). Ion pair quantification from differentially expressed peptides in a validation set of sera from 86 CHB patients was done with multiple reaction monitoring (MRM). RESULTS: 21 differentially represented peptide peaks were found through LC-MS/MS. Ion pairs generated from eleven of these peptides (m/z < 800) were quantified by MRM. Summed peak area ratios of 6 ion pairs from peptide m/z 520.3 (176.1, 353.7, 459.8, 503.3, 351.3, 593.1), which was identified as dihydroxyacetone kinase (DAK) fragment, decreased from mild to advanced stages of fibrosis or inflammation. Area Under Receiver Operating Characteristic Curves (AUROCs) of five ion models discriminating fibrosis degrees were 0.871 ~ 0.915 (S2-4 versus S0-1) and 0.804 ~ 0.924 (S3-4 versus S0-2). AUROCs discriminating inflammation grades were 0.840 ~ 0.902 (G2-4 versus G0-1) and 0.787 ~ 0.888 (G3-4 versus G0-2). The diagnostic power of these models provides improved sensitivity and specificity for predicting disease progression as compared to aspartate aminotransferase to platelet ratio index (APRI), FIB-4, Forn's index and serum DAK protein. CONCLUSIONS: The peptide fragment (m/z 520.3) of DAK is a promising biomarker to guide timing of antiviral treatment and to avoid liver biopsy in compensated CHB patients.

Serum proteomic MRM identify peptide ions of transferrin as new fibrosis markers in chronic hepatitis B.

BACKGROUND AND AIM: Because of the limitations of liver biopsy, reliable non-invasive serum biomarkers of liver fibrosis are needed. The aim of this study was to identify such markers by the use of serum proteomics in chronic hepatitis B (CHB). METHODS: Two-dimensional gel electrophoresis (2-DE) was used to identify differentially expressed protein spots in sera from 40 CHB patients [20 with mild fibrosis (S0-S1) and 20 with severe fibrosis (S3-S4)]. Mass spectrometry (MS) based multiple reaction monitoring (MRM) was used to quantify peptide ions of differential protein spots in another set of sera from 86 CHB patients with different liver fibrosis (S0-S4). RESULTS: Seven differentially expressed protein spots were found by 2-DE. Fourteen peptide ions of seven target protein spots were quantified by MS-based MRM. Summed peak areas ratio (SPAR) values of peptide ions from protein spot 1, 4 and 8, identified as apo serum transferrin, complement component C3c and transferrin, were significantly different from non-fibrosis (S0) to fibrosis stage 4. AUROCs of models established by peptide ions (protein spot 1, 4, 8) and model consisting of a combination of all ions were 0.848∼0.966 (S2-S4 versus S0-S1) and 0.785∼0.875 (S3-S4 versus S0-S2). Only the peptide ions model of transferrin had better sensitivity and specificity for predicting fibrosis stages than did aspartate aminotransferase-to-platelet ratio index (APRI), FIB-4 and Forn's index. CONCLUSIONS: Serum peptide ions of transferrin, detected by proteomic MRM, are new and promising biomarkers for staging liver fibrosis in CHB patients.

[Plasma proteome analysis of chronic hepatitis B patients with different stages of liver fibrosis].

OBJECTIVE: To identify plasma biomarkers with specific relation to the various liver fibrosis stages that can be used to assess hepatitis B virus (HBV)-infected patients for non-invasive liver fibrosis and to evaluate the prognosis of the liver fibrosis. METHODS: Plasma samples were collected from 80 HBV-positive patients at the Hepatitis Department of Shanghai Public Health Clinical Center between September 2008 and January 2011. The samples were grouped according to the patient's stage of hepatic fibrosis determined by liver biopsy: S0-1 (n = 40), S2-3 (n = 20), and S4 (n = 20). Each plasma sample was processed to remove the two most abundant proteins, albumin and immunoglobulin G (IgG), and then resolved by two-dimensional (2D) electrophoresis. ImageMaster 2D analysis software was used to identify differentially expressed proteins related to the liver fibrosis stages. After trypsin digestion, the differential proteins were identified by online reversed-phase nano-flow liquid chromatography coupled with electrospray ionization ion trap mass spectrometry (MS). RESULTS: The patients in the three groups were not significantly different in age (range: 30-50 years; P = 0.053) or sex (x² = 0.155, P = 0.926). Low-abundance proteins were efficiently enriched by the albumin/IgG depletion method. Fourteen differentially expressed proteins were detected among the S0-1, S2-3 and S4 groups, all of which were identified by tandem MS and included fibrinogen gamma chain, haptoglobin, complement C3, Ig kappa chain C region, and apolipoprotein A-I. CONCLUSION: Plasma proteomic analysis of chronic hepatitis B patients identified a panel of differentially expressed proteins related to different stages of liver fibrosis. These proteins may represent diagnostic and prognostic biomarkers of HBV-related hepatic fibrosis.

[Proteomic analysis of plasma membrane of HBV transgene mice livers].

OBJECTIVE: To study liver plasma membrane (PM) proteins affected by hepatitis B virus (HBV), and to study the molecular mechanism of HBV infection through plasma membrane proteome analysis of transgene mice livers. METHODS: Plasma membrane was purified using second antibody superparamagnetic beads that combine subcellular fractionation and immunoisolation strategies. Western blot was used to verify the purification of plasma membrane and the expression of HBV envelope protein. The proteins from plasma membrane were extracted, quantified and separated by two-dimensional electrophoresis. The gels were stained by silver nitrate or Coomassie Blue and analyzed by Imagemaster software. The different expressed proteins were identified by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF-TOF MS). RESULTS: The plasma membrane of mice livers was enriched 3-fold and the contamination from mitochondria was reduced 2-fold in comparison with the density gradient centrifugation method. HBV envelope protein was found only in HBV transgene mice livers. More than 500 protein spots were separated in Coomassie Blue stained gels. Twenty-six proteins with two-fold differences were found through Imagemaster software analysis. Seven proteins were identified. CONCLUSION: An optimized plasma membrane purification method was developed; protein profile of liver plasma membrane changed in the transgene mice livers; some proteins related to HBV infection were found, and this work may be helpful in the research of the molecular mechanism of HBV infection.

[Multicenter study on screen method for gestational diabetes].

OBJECTIVE: To investigate the feasibility of using random blood glucose to screen gestational diabetes mellitus (GDM). METHOD: The random blood glucose was determined in 1 038 pregnant women between 24 and 32 gestational weeks. Then 50 gram glucose challenge test (50 g GCT) was performed followed immediately. Finally, 75 gram oral glucose tolerance test (75 g OGTT) was done without dietary control for 3 days. If two values of four were abnormal, GDM was diagnosed. Impaired glucose tolerance (IGT) was diagnosed if only one value was abnormal or the 2nd hour value ranged from 120 to 164 mg/dl. RESULTS: (1) The determination of the three steps was completed in 948 cases. Among them, 42 cases (4.4%) were GDM, 372 cases (39.2%) were IGT and other 534 cases were normal. (2) In the normal group, the random blood glucose were different in fasting and postprandial times. No difference was found among blood glucose values determined of 50 g GCT at different times except that the value of 50 g GCT 1 hour postprandial was higher than the value at other times. There was no significant association between random blood glucose and 50 g GCT. (3) The sensitivity and specificity were 50.0% and 67.7%, when IGT was diagnosed using the cut point of 6.4 mmol/L (115 mg/dl) of random blood glucose, which was similar with 51.1% of sensitivity and 71.2% of specificity when using >or= 7.8 mmoL/L (140 mg/dl) as the cut point of 50 g GCT. If 6.4 mmol/L (115 mg/dl) was used as the cut point in GDM group the sensitivity would be 80.0%, which was much higher than that of IGT group and the specificity was 61.2%. In this study, if the value of >or= 8.3 mmoL/L (150 mg/dl) was used as the cut-point of 50 g GCT to screen the GDM, the sensitivity decreased only 2.0% while the specificity increased more than 10.0%. CONCLUSIONS: (1) The determination of random blood glucose to screen GDM couldn't replace the 50 g GCT, but it can be used as a complemental method and can be used repeatedly at any gestational age and convenience the pregnant women and the doctors. (2) The value of 8.3 mmol/L (150 mg/dl) was used as the cut-point of 50 g GCT, the specificity would be increased and the requirement for OGTT would be lowered markedly, which would reduce economic and psychological stress.