Automatic Biochemical Analyzer Research Articles

Association between fluoride exposure and the risk of serum CK and CK-MB elevation in adults: a cross-sectional study in China

BackgroundThis study aimed to investigate the relationship between urinary fluoride concentration and myocardial disease.MethodsThis is a cross-sectional study that was conducted in three villages in Wenshui County, Shanxi Province. A total of 737 villagers were included in this analysis. Urinary fluoride was detected using a fluoride-ion selective electrode. Myocardial enzymes were detected using an automatic biochemical analyzer. Myocardial ischemia and arrhythmia were diagnosed using 12-lead electrocardiogram.ResultsThe median level of urinary fluoride concentration was 1.32 mg/L. Urinary fluoride was associated with serum creatine kinase (CK) elevation (odds ratio [OR] = 1.39 [95% confidence interval (CI)]: 1.09–1.78) and CK isoenzyme (CK-MB) elevation (OR = 1.49 [95% CI: 1.12–1.97]). Stratified analysis revealed that urinary fluoride concentration was associated with CK elevation in villagers under the age of 60 years (OR = 1.80 [95% CI: 1.26–2.59]). This study found that there was a positive association between urinary fluoride concentration and the risk of CK-MB elevation in participants under the age of 60 years(OR = 2.18 [95% CI: 1.39–3.42]), those who were of female gender (OR = 1.53 [95% CI: 1.07–2.19]), those who were overweight/obese (OR = 1.96 [95% CI: 1.28–2.99]), those who had central obesity (OR = 1.59 [95% CI: 1.12–2.25]), consumed alcohol (OR = 1.49 [95% CI: 1.09–2.05]), and smoked (OR = 1.50 [95% CI: 1.10–2.04]).ConclusionOur study suggests that fluoride exposure is associated with the risk of serum CK and CK-MB elevation; however, it is not associated with myocardial ischemia, arrhythmia, serum lactate dehydrogenase (LDH), serum alpha-hydroxybutyrate dehydrogenase (α-HBD), or serum aspartate aminotransferase (AST). Further investigations are needed to substantiate our findings and explore the potential underlying mechanisms.

Dapagliflozin combined with metformin improves blood glucose, bone metabolism and bone mineral density in elderly patients with type 2 diabetes mellitus complicated with osteoporosis.

The incidence of type 2 diabetes mellitus (T2DM) complicated with osteoporosis (OP) (T2DM-OP) is growing. Dapagliflozin and metformin are commonly prescribed to manage glycemic levels in T2DM patients. We investigated the clinical efficacy of combining dapagliflozin with metformin in elderly patients with T2DM-OP. Totally 144 T2DM-OP patients were prospectively enrolled and allocated into two groups: the Metformin and Dapagliflozin + Metformin groups. Each group received treatment for 12 months. Fasting peripheral blood samples were collected before and after 12 months of treatment. Glycemic parameters and bone metabolic parameters were measured using oral glucose tolerance test, automatic biochemical analyzers, or liquid chromatography. Bone mineral density (BMD) changes at lumbar vertebrae (L1-4), femoral neck (FN) and total hip (TH) were assessed using dual-energy X-ray bone mineral densitometry. Pain severity was evaluated using the visual analog scale (VAS). The total effective rate, fracture incidence, and adverse reaction rate were also evaluated. After 12 months, both groups showed improvements in glycemic parameters, bone metabolic parameters, and BMD at L1-4, FN, and TH, and reductions in VAS scores. The Dapagliflozin + Metformin group exhibited more significant improvements. The overall effective rate was higher and fracture incidence, was lower in Dapagliflozin + Metformin group, with comparable rates of adverse reactions and safety profiles between the two groups. Taken together, treatment with a combination of dapagliflozin and metformin led to improvements in blood glucose levels, bone metabolism, and BMD in elderly patients with T2DM-OP, demonstrating superior efficacy and safety compared to metformin monotherapy.

Study on Hematological and Biochemical Index of Blood and Vitreous Humor in the Celestial-Eye Goldfish

In this study, we conducted an exploration of blood collection, hematological and biochemical comparative analysis of the unique vitreous humor and blood of the celestial-eye goldfish (CE) (Carassius auratus), and obtained hematological and biochemical indicators for healthy celestial goldfish blood. After exploring the optimal method for collecting blood and ocular fluid, blood and vitreous humor samples were drawn from healthy CE, and an automatic hematology analyzer was used to measure 17 physiological indicators in the blood and vitreous humor. Additionally, 20 biochemical indices were measured by an automatic biochemical analyzer. Experiments were also conductedto evaluate the effect of blood and vitreous humor collection on the subsequent survival rate of the CE. The results showed that the blood of CE exhibited values for 17 hematological indicators, including red blood cells (mean value: 2.19 × 1012/L), white blood cells (mean value: 62.21 × 109/L), hemoglobin (mean value:138.25 g/L), and no eosinophils or basophils. We also found that the levels of aspartate amino transferase (AST) and glutamyl transpeptidase (GGT) in the vitreous humor of CE were significantly higher than those in the serum (p < 0.01). No cells were detected in the vitreous humor of CE, and there was no significant difference in survival rate compared to normal goldfish after extracting blood and vitreous humor from CE (p > 0.05). This study provides fundamental data for the cultivation of healthy goldfish, the development of goldfish hematology, and further research on the development and formation of CE’s unique eye traits.

Effectiveness of a Novel PLA2R1 Knock-In Rat Model in Repairing Renal Function Damage.

Phospholipase A2 receptor 1 (PLA2R1) exists in many animals and plays an important role in membranous nephropathy. In this study, we aimed to evaluate a PLA2R1 knock-in rat model with repaired kidney function to study the molecular mechanisms of membranous nephropathy. We constructed the PLA2R1 knockout [PLA2R1(-)] model and PLA2R1 knock in [PLA2R1(+)] model in rats. Consistent complement C3 and IgA expression was confirmed through colocalization studies. Urinary biochemical indicators were performed using Automatic Biochemistry Analyzer. The complement C3, IgG, and Nephrin were detected by immunofluorescence assay. The expression levels of complement C3, IgA, and PLA2R1 were detected by western blot. The differential expression proteins (DEPs) between control and PLA2R1(+) models were detected by liquid chromatography with tandem mass spectrometry. The PLA2R1(-) model showed proteinuria, complement C3 aggregation, and IgA and IgG deposition in the glomerulus. Comparing with the PLA2R1(-) model, the PLA2R1(+) model, the deposition of complement C3 and IgA in the glomerulus did not completely disappear, and IgG expression weakened. Moreover, the absolute value of urinary protein was much lower in the PLA2R1(+) model than in the PLA2R1(-) model, and some of the humanized PLA2R1 gene fragments repaired some of the kidney functions. Humanized PLA2R1-insertion in rats can repair part of the renal function and reduce proteinuria, which will help in studying the molecular mechanisms of membranous nephropathy, as well as the entire membranous nephropathy-related system and complement activation signaling pathway.

Zhuyu pill attenuates metabolic-associated fatty liver disease by regulating macrophage polarization through TLR4 signaling pathway.

Metabolic-associated fatty liver disease (MAFLD) is the leading chronic liver disease globally, impacting a large segment of the population. The Zhuyu Pill (ZYP), a traditional Chinese remedy, has been clinically used for treating MAFLD, with its effectiveness demonstrated in both human patients and animal models. However, the underlying mechanisms of how ZYP addresses MAFLD still require further investigation. This study investigated the molecular mechanism of ZYP in treating MAFLD through both in vivo and vitro methods. A murine MAFLD model was induced by a high-fat, high-fructose diet for 12 weeks. ZYP was administered for 4 weeks, with fenofibrate serving as a positive control. Indicators of lipid metabolism in serum and liver tissue were detected by automatic biochemical analyzer and ELISA, respectively. Histopathological evaluation of liver sections was performed using HE and oil red O staining. Transcriptomics was employed to further investigate the therapeutic mechanism of ZYP in MAFLD. Additionally, macrophages and their polarization in the liver were analyzed using ELISA, flow cytometry, immunohistochemistry, and immunofluorescence (IF). Candidate proteins and pathways were validated in vivo and in vitro by western blotting and IF. Validation of the pathway was performed in vitro using inhibitors and co-culture strategies. ZYP significantly improved obesity and hepatic steatosis in MAFLD mice, reducing body/liver weight and regulating lipid metabolism indicators in serum and liver tissue. Bioinformatics analysis of transcriptomic data highlighted lipid metabolism regulation and inflammation control as key effects of ZYP in treating MAFLD. The in vivo experimental results showed that ZYP inhibited M1 polarization of macrophages (pro-inflammatory) and promoted M2 polarization of macrophages (anti-inflammatory) in MAFLD mice. Further in vivo and vitro experiments indicated that ZYP competes with LPS to bind to Toll-like receptor 4 (TLR4), suppressing M1 polarization in liver macrophages, and improving MAFLD. The in vitro co-culture system also confirmed that ZYP reduces liver lipid deposition by modulating M1 macrophage polarization. ZYP alleviates MAFLD by inhibiting M1 polarization of liver macrophages, indicating that ZYP may be a promising treatment for MAFLD. Its mechanism of action is to inhibit the TLR4/MyD88/TRAF6 signaling pathway, modulate macrophage polarization, and improve inflammatory response.

Effect of Dietary Oils with Different Fatty Acid Compositions on Serum Lipid and Gut Microbiota of Rats.

The effects of three dietary oils (rapeseed oil, camellia oil, linseed oil) with different fatty acid compositions on the growth performance, digestion and gut microbiota of SD rats after 8 weeks of feeding were studied. The serum metabolic index and liver histomorphology of rats were measured using an automatic biochemical analyzer and light microscope. Furthermore, 16S rDNA amplicon sequencing technology was used to analyze the gut microbiota. It was found that these differences in fatty acid composition had no significant effect on body fat and liver tissue. However, after digestion, the rapeseed oil group showed lowest triglyceride content (1.22 ± 0.15) and a lower LDL/HDL ratio (0.41 ± 0.02). For gut microbiota distribution, the linseed oil group showed a higher Firmicutes/Bacteroides ratio (6.11 ± 0.54) and a high proportion of Lactobacillus. These data indicate that both the unsaturated fatty acid content and n-3 unsaturated fatty acids collectively had an effect on digestion metabolism, and the influence order may be n-3 unsaturated fatty acids > unsaturated fatty acid content.

Electroacupuncture preconditioning alleviates podocyte injury via PTEN/PI3K pathway in type 2 diabetes rats

To observe the effect of electroacupuncture (EA) preconditioning on podocytes and phosphatase and tension homologous protein (PTEN)/phosphoinositide-3-kinase (PI3K) signaling pathway in rats with type 2 diabetic kidney injury, so as to explore its potential mechanisms. Fifty male Wistar rats were randomly divided into control, model, EA, inhibitor, and sham EA groups, with 10 rats in each group. Diabetes model was established by high-fat and high-sugar diet and intraperitoneal injection of streptozotocin (40 mg/kg). For rats in the EA group, EA (2 Hz, 1 mA) preconditioning was applied to "Guanyuan" (CV4), "Zhongwan" (CV12), and bilateral "Zusanli" (ST36), "Fenglong" (ST40) for 15 min, once every other day for 8 weeks. And rats in the sham EA group were given acupuncture at the subcutaneous areas of the same acupoints without EA stimulation. Rats of the inhibitor group were given intraperitoneal injection of BPV (HOpic, 0.6 mg/kg, 0.5 mg/mL) combined with the same EA stimulation as EA group once a week for 8 weeks. The blood glucose level of rats was recorded. Urinary albumin (ALB) content in rats was detected by ELISA. The contents of serum urea (Urea), creatinine (Crea), total cholesterol (TC) and triglyceride (TG) were detected by automatic biochemical analyzer. HE staining, PAS staining and Masson staining were used to observe the pathological changes of renal tissues. The ultrastructural changes of podocytes were observed by transmission electron microscopy. The relative expression levels of Synaptopodin, microtubule-associated protein light chain 3 Ⅱ(LC3-Ⅱ), PTEN and PI3K proteins in kidney were detected by Western blot. Compared with the control group, the blood glucose, content of serum ALB, Urea, Crea, TC, TG and renal PI3K protein expression in the model group were significantly increased (P<0.01), while the expression levels of renal Synaptopodin, LC3-Ⅱ and PTEN proteins were decreased (P<0.01). Compared with the model group, the blood glucose, content of serum ALB, Urea, Crea, TC, TG and renal PI3K protein expression in the EA group decreased (P<0.01, P<0.05), while the expression levels of renal Synaptopodin, LC3-Ⅱ and PTEN proteins increased (P<0.01). In comparison with the EA group, the blood glucose, ALB, Urea, Crea, TC, TG content and PI3K protein expression level were increased (P<0.05, P<0.01), while the expression levels of Synaptopodin, LC3-Ⅱ and PTEN proteins were decreased (P<0.01) in the inhibitor group；whereas, the blood glucose, ALB, Urea, TC, TG content and PI3K protein expression level were increased (P<0.05, P<0.01), while the expression levels of Synaptopodin, LC3-Ⅱ and PTEN proteins were decreased (P<0.01) in the sham EA group. HE staining and PAS staining showed glomerular hypertrophy and glomerular glycogen deposition；and Masson staining displayed an enhancement of glomerular fibrosis；and electron microscope revealed foot process fusion, basement membrane thickening and autophagosomes reduction in the rat' s kidney of the model, inhibitor and sham EA groups, which were relatively milder in the EA group. EA promotes the expression of PTEN gene-encoded protein in the kidneys of type 2 diabetic rats, thereby inhibiting the activation of PI3K, increasing the autophagy level and protecting the podocytes.

Effects of alpha-lipoic acid on productive performance, immunity, antioxidation and serum metabolism in Holstein cows

Abstract The aim of this study was to investigate the antioxidant effect of alpha-lipoic acid (α-LA) in dairy cows and its metabolic mechanism. Thirty Holstein cows weighing 550 ± 25 kg, 200 ± 15 days of lactation and calving 2–3 times were randomly divided into three groups, ten cows in each group. Different doses of α-LA were added based on body weight: 0 (CTL), 30 (LA-L) and 60 (LA-H) mg/kg per head per day; 7 days adaptation period, 30 days formal period. Milk production was recorded daily during the test period. Milk and blood samples were collected on the last day. ELISA kits and automatic biochemical analyser were used to detect the indicators in blood; serum metabolites were detected and analysed by non-target metabolomics. The results of the study showed that the addition of α-LA significantly increased milk yield; blood concentrations for HDL, triglyceride, cortisol and triiodothyronine were significantly elevated; and levels of glutathione reductase and nitric oxide synthase were significantly reduced in LA-L group as compared to CTL group. The concentrations of IL-1β, IL-2, TNF-α, IgG and IgA were significantly higher after supplementation with α-LA. Metabolomics analysis revealed 13 and 15 differential metabolites each in positive or negative modes. Methylmalonic acid levels were significantly higher following α-LA supplementation compared to CTL group, as were D-lactose, D-maltose and oleanolic acid levels in LA-L group. In summary, α-LA can enhance milk production, improve antioxidant capacity and immunity, and is more beneficial for animal production and economic benefits at 30 mg/kg.

. Diagnostic value of HMGB-1 and Acetylcholinesterase in Assessing the Prognosis of Patients with Acute Pancreatitis

Aim: Acute pancreatitis (AP) is a disorder of tissue digestion caused by abnormal activation of pancreatic enzymes, which may lead to multi-organ failure and ultimately death as the disease progresses. How to quickly and accurately evaluate the progression of AP has always been a hotspot and difficulty in clinical research. Methods: Sixty-four AP patients admitted to our hospital from August 2022 to June 2023 and 60 concurrent healthy people were selected as the research subjects, with AP patients as the observation group and healthy people as the control group. Using kits from Wuhan Fine Biotechnology Co., Ltd., an enzyme-linked immunosorbent assay (ELISA) was carried out to measure HMGB-1 and AChE levels. In addition, an automatic biochemical analyzer was utilized to quantify the levels of nutritional proteins albumin (ALB), transferrin (TRF), and total protein (TP) in the observation group. Results: Compared with the control group, the HMGB-1 in the observation group was higher while the AChE was lower (P&lt;0.05). ROC curve analysis showed that when HMGB-1 was less than 7.25 mg/L, the sensitivity and specificity for diagnosing AP were 89.06% and 51.67%, respectively (P&lt;0.05); when AChE was less than 6.18 U/L, the sensitivity for diagnosing AP was 43.75% and the specificity was 90.00% (P&lt;0.05). Using the joint detection Log(P)=-1.736+(0.645×HMGB-1)+(-0.454×AChE), it was found that HMGB-1+AChE had a sensitivity of 48.44% and a specificity of 88.33% for the diagnosis of AP (P&lt;0.05, cut-off&gt;0.639,) Conclusion: In this study, we found elevated HMGB-1 expression in AP and reduced acetylcholinesterase (AChE), suggesting that both of them may be involved in the occurrence and development of AP. Meanwhile, the two demonstrated excellent diagnostic efficacy in the onset and poor prognosis of AP, and were closely related to AP progression, confirming their potential as new clinical indicators for AP. Due to the self-digestion by pancreatic enzymes, the nutritional metabolism of AP patients is disrupted, so protein loss is also a focus of attention. In this study, we also found a negative correlation of nutritional protein levels with HMGB-1 in AP patients and a positive correlation between them with AChE, further supporting the relationship between the two and AP progression. The findings can lay a reliable foundation for future research on the diagnosis and treatment of AP based on HMGB-1 and AChE.

Study of plasma essential element concentrations to explore markers of acute myocardial infarction.

Essential element concentrations in biological samples may be related to the pathogenesis of various diseases. Previous studies have reported that serum iron (Fe), zinc (Zn), and copper (Cu) were related to acute myocardial infarction (AMI). However, the differences in element concentrations between AMI and other cardiac disease has not been investigated. In this study, differences in plasma Fe, magnesium (Mg), Zn, Cu, calcium (Ca), inorganic phosphorus (P), and cardiac troponin T (TnT) levels in heart disease patients (AMI, angina, heart failure, and chest pain) were investigated to explore potential markers of AMI. Fe, Mg, Zn, and Cu concentrations were assayed by using a Metallo Assay kit; Ca and inorganic P were determined by using an automatic biochemical analyzer; and cardiac TnT levels were assayed by using enzyme-linked immunosorbent assay. Plasma TnT levels were higher in AMI than in other heart diseases and were negatively correlated with Cu and Ca. Fe, Cu, and inorganic P levels were within the normal range, while Mg and Ca levels were lower, and Zn levels were higher than the normal range in heart disease patients. Except Mg, no significant differences in element levels were observed among heart diseases: Mg levels were significantly higher in AMI than in heart failure. These results suggest that lower Cu and Ca levels and a higher Mg level compared with other heart diseases may be a marker of AMI.

Tanshinone Ⅱ_A exerts anti-hepatocellular carcinoma effects by inhibiting oxidative stress via PI3K/Akt and Nrf2/HO-1 signaling pathway

This study aims to investigate the mechanism of tanshinone Ⅱ_A(Tan Ⅱ_A) in protecting mice from diethylinitrosamine(DEN)/carbon tetrachloride(CCl_4)/ethanol(C_2H_5OH)-induced hepatocellular carcinoma(HCC) and HepG2 cells from hydrogen peroxide(H_2O_2)-induced oxidative damage via the phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt) and nuclear factor E2-related factor 2(Nrf2)/heme oxygenase 1(HO-1) signaling pathways. Sixty male C57BL/6J mice were grouped as follows: control, model, low, medium, and high-dose(10, 20, 40 mg·kg~(-1), respectively) Tan Ⅱ_A, and colchicine(0.2 mg·kg~(-1)), with 10 mice in each group. The HCC model was established with DEN/CCl_4/C_2H_5OH for 20 weeks, and the mice were then euthanized for collection of blood and liver specimens. A protein-protein interaction(PPI) network of the targets of Tan Ⅱ_A in the prevention of HCC was constructed. HepG2 cells were treated with 150 μmol·L~(-1) H_2O_2 for the modeling of oxidative stress. The cell counting kit-8(CCK-8) was used to assess the effects of different concentrations(1, 2, 4 μmol·L~(-1)) of Tan Ⅱ_A on the relative viability of cells. Hematoxylin-eosin(HE) staining was used to observe the pathological changes in the liver tissue, and an automatic biochemical analyzer was used to measure the levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) in the mice serum. Relevant assay kits were used to measure the levels of reactive oxygen species(ROS), malondialdehyde(MDA), superoxide dismutase(SOD), glutathione(GSH), and glutathione peroxidase(GSH-Px) in the liver and cells. Immunohistochemistry, immunofluorescence assay, and Western blot were employed to determine the expression of phosphorylated PI3K(p-PI3K), PI3K, phosphorylated Akt(p-Akt), Akt, Nrf2, and HO-1. Compared with the control group, the model group exhibited typical pathological manifestations of liver cancer in the liver tissue, with elevated levels of ALT and AST in the serum, risen levels of ROS and MDA in the liver, and lowered levels of SOD and GSH in the liver. Compared with the control group, the HepG2 cells treated with H_2O_2 showed significantly decreased activities of SOD and GSH-Px and increased ROS and MDA levels. In the liver tissue and HepG2 cells, the modeling up-regulated the expression of p-PI3K and p-Akt while down-regulating the expression of Nrf2 and HO-1. Compared with the model group, different doses of Tan Ⅱ_A reduced the levels of ALT and AST in the serum, elevated the levels of SOD, GSH, and GSH-Px in the liver tissue and cells, and lowered the ROS and MDA levels, which indicated significant alleviation of oxidative stress. The PPI network showed that Akt was a core target of the PI3K/Akt and Nrf2/HO-1 signaling pathways. Western blot, immunohistochemistry, and immunofluorescence assay results indicated that Tan Ⅱ_A promoted the expression of Nrf2 and HO-1 while inhibiting the phosphorylation of PI3K and Akt. In conclusion, Tan Ⅱ_A may delay the progression of HCC by inhibiting oxidative stress via the PI3K/Akt and Nrf2/HO-1 signaling pathways.

Clinical Value of Circ-PNPT1 on Adverse Pregnancy Outcomes of Patients with Gestational Diabetes Mellitus.

Several circular RNAs are associated with important pathophysiological characteristics of gestational diabetes mellitus (GDM). This study intended to measure the expression of circ-PNPT1 in sera of GDM patients and to expound on its values on pregnancy outcomes. Totally 104 GDM patients and 71 healthy controls were recruited. The expression pattern of serum circ-PNPT1 was measured by reverse transcription-quantitative polymerase chain reaction. The diagnostic efficacy of circ-PNPT1 and fasting blood glucose (FBG) on GDM was evaluated by receiver operating characteristic (ROC) analysis. Parameters of glycolipid metabolism were determined using automatic biochemical analyzers. The correlation between circ-PNPT1 and glycolipid metabolism parameters was analyzed using Pearson analysis. GDM patients were divided into a high expression group and a low expression group based on the median value of circ-PNPT1 expression. Curves of adverse neonatal outcomes were drawn by Log Rank analysis. GDM patients exhibited higher circ-PNPT1 expression than healthy controls. The area under the ROC curve of circ-PNPT1 diagnosing GDM was 0.9184 and the cut-off value was 1.435 (90.38% sensitivity, 85.92% specificity). Serum circ-PNPT1 expression was positively correlated with FBG, total cholesterol, and triglyceride in GDM patients. Neonates born to GDM patients with high circ- PNPT1 expression were prone to adverse outcomes. Circ-PNPT1 was highly-expressed in the sera of GDM patients. Circ-PNPT1 affected glycolipid metabolism and its expression had certain reference values on adverse pregnancy outcomes.

M1 polarization of hepatic macrophages in cows with subclinical ketosis is an important cause of liver injury.

Subclinical ketosis (SCK) is highly prevalent and easily overlooked, with insidious and slow progression of hepatic injury, often characterized by an imbalance in immune homeostasis. In nonruminants, macrophage polarization plays an important regulatory role in hepatic lipid accumulation, fibrosis and inflammatory processes. Thus, we aimed to investigate the status of hepatic macrophage polarization in SCK cows and to corroborate its association with liver injury and inflammation. Twelve Holstein dairy cows (parity, 2-4) were selected, and liver biopsy and blood were collected on the second week postpartum (10-14 d days in milk). On the basis of serum BHBA concentrations., selected cows were categorized into healthy (n = 6; BHBA <1.0 mM) and SCK (n = 6; 1.2 mM ≤ BHBA < 3.0 mM) groups. Serum biochemical parameters were measured using an automatic biochemical analyzer, which indicated higher serum levels of BHBA and NEFA and an upregulation of liver injury indicators (AST, ALT, TP, GLB) in SCK cows compared with healthy cows. ELISA assays revealed that SCK cows displayed systemic low-grade inflammation, as demonstrated by increased serum levels of HP, SAA, TGF-β, IFN-γ, and IL-1β. Liver biopsies revealed pathological histological alterations, hepatic inflammation, and macrophage polarization status. Oil red staining indicated steatosis, while Sirius red staining demonstrated mild extracellular matrix deposition in the liver of SCK cows. The expression of inflammatory response-related proteins (TLR4, p-NFκB, p-I-κB, NLRP3, and Caspase-1) was elevated in the liver of SCK cows, with the increased mean fluorescence intensity of NFκB further confirming the activation of the inflammatory pathway. Furthermore, the levels of pro-inflammatory factors, TNF-α and IFN-γ, were elevated in the tissue homogenate. Macrophage phenotypic changes in SCK cows were further explored based on the results of liver injury and inflammation. Compared with healthy cows, the protein and mRNA abundance of the macrophage marker CD68 in the liver of SCK cows was higher, along with an increased mean fluorescence intensity of CD68. SCK cows also exhibited reduced mRNA expression of the Kupffer cell marker CLEC4F and elevated chemokine levels (CXCL1 and CCL2). As evidenced by greater protein and mRNA abundance of macrophage M1 polarization markers (iNOS, IL-1β, CD86, IL-6, IL-12b, and CCL3), higher fluorescence intensity of iNOS and CD86, and an increased number of CD68+/CD86+-positive cells observed via immunofluorescence, the macrophage polarization phenotype in the liver of SCK cows was predominantly M1. In contrast, the protein and mRNA abundances of M2 polarization markers (CD206, IL-10, and Arg1) were lower in SCK cows, accompanied by a reduced fluorescence intensity of CD206 and a lower number of CD68+/CD206+-positive cells. Overall, the present study revealed that the number of macrophages in liver is enhanced during subclinical ketosis and is dominated by pro-inflammatory macrophages (M1 macrophages). This could partly explain the increased risk of steatosis, fibrosis, and inflammatory response processes in these cows.

Uricase-Expressing Engineered Macrophages Alleviate Murine Hyperuricemia.

Background: Uricase, or urate oxidase (Uox) is a key enzyme in uric acid (UA) metabolism and has been applied in clinical treatment of human hyperuricemia (HUA). However, the current clinically applied uricases, despite their potent urate-lowering capacity, tend to form anti-drug antibodies because of their immunogenicity, leading to increased risk of anaphylaxis, faster drug clearance and reduced or even complete loss of therapeutic effect, limiting their clinical application. In this study, we constructed engineered macrophages that stably expressed uricase, which might serve as a promising alternative to the direct injection of uricases. Materials and Methods: Engineered macrophages RAW264.7 cells were injected intravenously to treat hyperuricemic KM mice. Serum uric acid and bio-indicators for renal and hepatic functions were detected by an automatic biochemical analyzer; inflammatory cytokines were determined by ELISA; the livers and kidneys of the mice were sectioned for histological examination. Results: The uricase-expressing macrophages reduced UA levels from 300 ± 1.5 μmol/L to 101 ± 8.3 μmol/L in vitro. And in an HUA mouse model established by gavage with yeast extract, intravenous injection of the engineered macrophages could reduce the serum uric acid (sUA) of mice to normal level on the 14th day of modeling, with a decrease of 48.6%, and the urate-lowering effect was comparable to that of the first-line clinical drug allopurinol. In terms of safety, engineered macrophages did not cause liver or kidney dysfunction in mice, nor did they induce systemic immune response. Conclusions: Using macrophages as a chassis to deliver uricase might be a new, safe and effective strategy for the treatment and control of hyperuricemia.

Mechanism of action of exercise regulating intestinal microflora to improve spontaneous hypertension in rats.

Hypertension is a prevalent cardiovascular disease. Exercise is widely recognized as an effective treatment for hypertension, and it may also influence the composition of the intestinal microflora. However, it remains unclear whether exercise can specifically regulate the intestinal microflora in the context of hypertension treatment. In this study, tail blood pressure in spontaneously hypertensive rats (SHR) was measured using a blood pressure meter after exercise intervention and fecal bacteria transplantation following exercise. Blood lipid levels were assessed using an automatic biochemical analyzer, and 16S rRNA sequencing was employed to analyze the intestinal microflora. Histological examinations of ileal tissue were conducted using HE and Masson staining. Intestinal permeability, inflammatory status, and sympathetic activity were evaluated by measuring the levels of diamine oxidase, D-lactic acid, C-reactive protein, interleukin-6, tumor necrosis factor-α, lipopolysaccharide, norepinephrine, angiotensin II, cyclic adenosine monophosphate, and cyclic guanosine monophosphate. Exercise was found to reduce blood pressure and blood lipid levels in SHR. It also improved the composition of the intestinal microflora, as evidenced by a reduced Firmicutes/Bacteroidetes ratio, an increase in bacteria that produce acetic and butyric acid, and higher Chao 1 and Shannon diversity indices. Furthermore, exercise reduced the thickness of the fibrotic and muscular layers in the ileum, increased the goblet cell/villus ratio and villus length, and decreased intestinal permeability, inflammatory markers, and sympathetic nerve activity. The intestinal microbial flora regulated by exercise demonstrated similar effects on hypertension. In conclusion, exercise appears to regulate the intestinal microflora, and this exercise-induced change in flora may contribute to improvements in hypertension in rats.

Interaction of polychlorinated biphenyls exposure, lifestyle and genetic risk in dyslipidemia

Abstract Background Polychlorinated biphenyls (PCBs) are endocrine-disrupting chemicals that have been associated with various adverse health conditions. However, the interaction effects between PCBs, lifestyle, and/or genetic susceptibility on dyslipidemias are still unknown. Methods Six serum PCBs including PCB-28, 101, 118, 138, 153, and 180 were determined in 3,845 participants from the Wuhan-Zhuhai cohort. Dyslipidemias, including hyper-total cholesterol (HyperTC), hyper-triglyceride (HyperTG), hyper-low density lipoprotein cholesterol (HyperLDL-C), and hypo-high density lipoprotein cholesterol (HypoHDL-C) were classified based on serum lipids levels, which were examined by an automatic biochemical analyzer. Besides, we constructed lipid-specific polygenic risk scores (PRS) and healthy lifestyle scores, and used Logistic regression models to explore the relationships of serum PCBs, lifestyle, and PRS with dyslipidemias. Results We observed that all six PCB congeners were positively associated with the prevalence of dyslipidemias to varying degrees. In particular, dose-response manners were observed between ΣPCB levels and odds ratios (ORs) of HyperTC, HyperTG, and HyperLDL-C, respectively. Besides, we found that PRSs and unhealthy lifestyles were both positively associated with dyslipidemias. For the combined effect of ΣPCB, lifestyle, and PRS, participants with high ΣPCB, unfavorable lifestyle, and high genetic risk had the highest odds of HyperTC (2.285; 1.533, 3.417), HyperTG (7.944; 5.169, 12.329), and HyperLDL-C (3.523; 2.311, 5.394) when compared with those with low ΣPCB, healthy lifestyle, and low genetic risk. Conclusions Our study provided evidence that PCB exposure potentially increases the prevalence of dyslipidemias, which may be exacerbated by genetic susceptibility and unhealthy lifestyles. Key messages • Our results highlighted the importance of reducing PCB exposure, adopting healthy lifestyles for the prevention of dyslipidemia. • Our work might facilitate the emergence of reducing or regulating PCB pollution to control blood lipids and the resultant dyslipidemia epidemic.

Effect of Particle Size on Physical Properties, Dissolution, In Vitro Antioxidant Activity, and In Vivo Hepatoprotective Properties of Tetrastigma hemsleyanum Diels et Gilg Powders.

Objective: The aim of this study was to analyze the effects of different particle sizes of Tetrastigma hemsleyanum Diels et Gilg (TDG) powders on physical properties, dissolution, in vitro antioxidant activity, and in vivo hepatoprotective properties. Methods: The particle size of TDG coarse powders (TDG-CP), TDG fine powders (TDG-FP), and TDG micro powders (TDG-MP) were measured by a laser particle size analyzer. The physical properties were measured according to the latest version of the Chinese Pharmacopoeia (Committee Chinese Pharmacopoeia 2020). The content of the total flavonoids, total polysaccharides, kaempferol-3-O-rutinoside, and rutin of TDG powders were determined using the NaNO2-Al (NO3)3 colorimetric method, the sulphate-phenol colorimetric method, and HPLC, respectively. In vitro dissolution and antioxidant activity were determined by the paddle method in phosphate buffer (pH 6.8) and the DPPH radical scavenging method, respectively. In addition, the liver tissue pathology was evaluated by hematoxylin and eosin staining (H&E), and the AST and ALT activities were measured by automatic biochemical analyzer. The superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) activities were measured by using commercial analysis kits. Results: As the particle size decreases, the fluidity of TDG powders decreased and the porosity increased. In addition, there were no significant differences in physical properties between low temperature pulverized powders and room temperature pulverized powders. The final dissolution rates of the four bioactive ingredients in TDG-MP were found to be 85.06%, 85.61%, 83.88%, and 83.26%, respectively, whereas in TDG-CP, the dissolution rates were significantly lower at 18.79%, 17.96%, 22.46%, and 24.35%. The EC50 values of TDG-CP, TDG-FP, and TDG-MP on DPPH scavenging activity were 0.82, 0.31, and 0.10 mg/mL, respectively. The AST and ALT activities of the TDG-FP group and the TDG-MP group were significantly decreased and the SOD, CAT, and GSH activities were significantly increased when compared with that of the model group. The inflammatory cell infiltration and vacuolar degeneration of liver cells in the TDG-FP group and the TDG-MP group were significantly improved. Conclusions: The particle size of TDG powders had a significant effect on the physical properties and in vivo bioactivity. TDG pulverized to a fine particle size or smaller is a promising approach for clinical applications with improved physicochemical and biological properties.

Serum Lipid Profile and Electrolytes Reference Intervals for Apparently Healthy Children and Adolescents in Addis Ababa, Ethiopia.

Accurate reference intervals generated from an apparently healthy population and stratified by crucial variables such as age and gender are required to guarantee appropriate interpretation of test results. Since there were no local reference intervals in the study area, the present study aimed to establish reference intervals on serum lipid profiles and electrolytes for children and adolescents in Addis Ababa. This community-based cross-sectional study was conducted from April to October 2019. Laboratory analysis was performed using the automatic biochemical analyzer Cobas 6000 (c501) from Roche. According to Clinical and Laboratory Standards Institute (CLSI) guidelines, reference intervals for lipid profile and electrolyte tests for apparently healthy children and adolescents were established. We used a non-parametric method to calculate the 2.5th and 97.5th percentiles with a 90% confidence interval. In children, the reference intervals for serum potassium, sodium, chloride, calcium, magnesium, and phosphate in mmol/L were 4.37-5.20, 137-145.50, 101.90-107.90, 2.34-2.70, 0.74-0.97, and 1.42-1.85, respectively; and for total cholesterol, triglycerides, low-density lipoprotein, and high-density lipoprotein, the respective values were 100.76-171.70, 44.16-126.36, 60.60-105.60, and 31.60-53.70 in mg/dL, for both genders. For adolescents, the reference intervals were 4.03-5.58, 137-146, 98.90-120.90, 2.39-2.70, 0.73-0.96, and 0.96-1.80 for serum potassium, sodium, chloride, calcium, magnesium, and phosphate in mmol/L, respectively; and 97.20-189.10, 40.50-143.60, 41.70-120.90, and 21.30-57.0 in mg/dL for total cholesterol, triglycerides, low-density lipoprotein, and high-density lipoprotein, respectively, for both genders. The established reference intervals in the current study revealed that both the lower and upper limits contradicted the manufacturer values as well as the available literature. The study also discovered significant gender differences in reference values for TC, TG, LDL-C, potassium, phosphate, and chloride in the adolescent age groups.

Emodin-based Regulation and Control of Serum Complement C5a, Oxidative Stress, and Inflammatory Responses in Rats with Urosepsis via AMPK/SIRT1.

Emodin, derived from Rheum officinale and aloe, is known for its diverse benefits such as anti-inflammatory, antioxidant, and antibacterial properties. Currently, the impact of emodin on urosepsis is unclear. This study aims to investigate the mechanism of action of emodin in urosepsis. Peripheral blood mononuclear cells (PBMCs) were purchased from Cloud-Clone Animal Inc. and treated with emodin. Cell viability and the lactate dehydrogenase (LDH) level were then assessed. In a separate experiment a urosepsis model was established in Sprague Dawley rats which were subsequently treated with emodin. The levels of oxidative stress-related factors, serum complements and inflammatory factors were measured using commercial kits. Blood urea nitrogen and serum creatinine levels were determined using a fully automatic biochemical analyzer. The levels of pro-inflammatory proteins and AMP-activated protein kinase (AMPK)/Sirtuin 1 (SIRT1) pathway-related proteins were evaluated via Western blot. PBMCs were unaffected by emodin concentrations below 60 μg/mL, and minimal LDH levels were detected in the cells. Emodin attenuated the effects of Escherichia coli and diminished the production of serum complements, oxidative stress-related proteins, and inflammatory factors in PBMCs. Notably, the effects of emodin were lessened by an AMPK pathway inhibitor. Additionally, emodin alleviated oxidative stress, complement system activation, inflammation, and kidney injury in urosepsis rats through the AMPK/SIRT1 signaling pathway. Emodin improved kidney damage in urosepsis rats by activating the AMPK/SIRT1 signaling pathway, which reduced oxidative stress, inflammation, and complement system activation.

Copper biomarkers and their relationship with dyslipidemia in women with obesity.

Adipose tissue dysfunction influences the development of dyslipidemias associated with obesity, however, the mechanisms are still unclear. In this sense, the literature highlights the role of copper in cholesterol synthesis, contributing to the increase in concentrations of this lipid fraction and consequently to the manifestation of dyslipidemia. The objective of the study was to investigate the relationship between copper parameters and lipid profile markers in women with obesity. This is a cross-sectional study involving women aged 20 to 50years divided into a case group (BMI ≥ 35kg/m2) and a control group (BMI between 18.5 and 24.9kg/m2). Copper concentrations in plasma and erythrocytes were determined by inductively coupled plasma optical emission spectrophotometry and ceruloplasmin activity by spectrophotometry. The lipid fractions were analyzed according to the enzymatic colorimetric method, using an automatic biochemical analyzer. Participants with obesity had elevated concentrations of copper in plasma and reduced concentrations in erythrocytes compared to the control group, but there was no significant difference in ceruloplasmin activity between the groups. The research does not identify a correlation between copper parameters and serum concentrations of lipid fractions, which does not allow inferring the role of copper in the manifestation of dyslipidemia in obesity.