Profile of Pathogenic Proteins and MicroRNAs in Plasma-derived Extracellular Vesicles in Alzheimer's Disease: A Pilot Study.

Protein and miRNA enrichment within extracellular vesicles (EVs) isolated from patients with Alzheimer's disease (AD) has been shown to have putative diagnostic value. However, whether a combination of both will be more advantageous is unknown. EVs were enriched from serum samples obtained from patients with sporadic AD (n = 13), mild cognitive impairment (MCI) (n = 10), vascular dementia (VaD) (n = 10), and healthy controls (HC) (n = 10). Expression of protein levels of beta-amyloid peptide (Aß1-42), total tau, P-T181-tau, and P-S396-tau and 18 microRNAs (miRNAs) in the EVs was performed by ELISA and qRT-PCR, respectively. Results were validated in an independent cohort of 18 subjects each by qRT-PCR assays. EV protein expression of Aß1-42, total-tau, P-T181-tau and P-S396-tau, were significantly different among AD, MCI and VaD. Hsa-miR-1306-5p, hsa-miR-342-3p, and hsa-15b-3p were all significantly downregulated in patients with AD compared to HC (P < 0.05), only hsa-miR-1306-5p expression was differentially expressed between AD, MCI, and VaD samples. Similarly, whereas all 14 miRNAs were significantly upregulated in patients with AD compared to HC, only hsa-miR-93-5p, hsa-miR-424-5p, and hsa-miR-3065-5p were differentially expressed when AD samples were compared to MCI and VaD samples. Even though the sample size was small, the results of the current pilot study indicates that hsa-miR-1306-5p, hsa-miR-93-5p, hsa-miR-424-5p, and hsa-miR-3065-5p, and expression of P-S396-tau in EVs might provide a combinatorial protein and miRNA signature to differentiate between HC, patients with MCI or VaD from patient with sporadic AD.

Ischemic Stroke in Young Adults of Northern China: Characteristics and Risk Factors for Recurrence.

BACKGROUND: Young adults accounted for 10-14% of ischemic stroke patients. The risk factors may differ in this population from elder patients. In addition, the factors associated with stroke recurrence in this population have not been well investigated. OBJECTIVE: The study aimed to investigate the characteristics and risk factors associated with recurrence of ischemic stroke in young adults. METHODS: Clinical data of 1,395 patients of age 18-45 years who were treated between 2008 and 2014 in 3 centers located in northern China was reviewed. The first onset of stroke was taken as the initial events and recurrent stroke as the end point events. The end point events, age, gender, duration after first onset of stroke, history of disease, National Institutes of Health Stroke Scale (NIHSS) score at admission, Trial of Org 10172 in Acute Stroke Treatment classifications of the cause of stroke and adherence to medication were recorded. These factors were analyzed and compared between recurrence and non-recurrence group. Information about recurrent stroke was collected through clinical (readmission to hospital with ischemic stroke) or telephone follow-up survey. Logistic regression was used to analyze the risk factors of recurrence. RESULTS: The most common causes of stroke were large vessel atherosclerosis and small vessel occlusion, followed by cardioembolism. NIHSS score at admission (OR 1.088; 95% CI 1.028-1.152; p = 0.004) were associated with recurrence. CONCLUSIONS: Vascular disease, especially premature atherosclerosis, is the major risk factor for ischemic stroke in the young adult population of northern China. Timely screening of the cause of stroke with severe NIHSS score needs further attention.

Plasma Phospholipids are Associated with Mild Cognitive Impairment in Type 2 Diabetic Patients.

BACKGROUND: Phospholipids and their metabolisms are closely allied to nosogenesis and aggravation of Type 2 diabetes mellitus and cognitive impairment. The aim of this study is to characterize the plasma levels of phospholipids in type 2 diabetes patients and type 2 diabetes patients with mild cognitive impairment (MCI), and to identify potential biomarkers of type 2 diabetes patients with MCI. METHODS: In this cross-sectional study, a total of 374 type 2 diabetes patients were prospectively enrolled. There were 103 patients with MCI and 271 patients without MCI. Plasma levels of lysophosphatidic acid (LPA) and phospholipids with solubility similar to that of LPA (PSS-LPA) were assayed. RESULTS: Plasma LPA and PSS-LPA levels were significantly higher in patients with MCI than those without MCI (P = 0.007, P ＜ 0.001). A logistic regression analysis indicates that elevated plasma PSSLPA was associated with increased risk of MCI in type 2 diabetic patients, with an OR of 1.87 (1.04- 3.47) after additional adjustment for hyperlipidemia, hypertension, High-sensitivity C-reactive protein, hemoglobin A1c, Intima-media thickness, ankle brachial index, and brachial-ankle pulse wave velocity. In type 2 diabetic patients with MCI, there were negative correlations between plasma LPA, PSS-LPA and the MoCA scores (r =﹣0.322, P < 0.01; r =﹣0.349, P < 0.001). Furthermore, plasma PSS-LPA exhibited a fair diagnostic value for MCI, with an area under the curve of 0.86. CONCLUSION: The level of plasma PSS-LPA may be an important biomarker for diagnosis of MCI.

Biomarkers of Renal Function in Type 2 Diabetic Patients with Cognitive Impairment.

Kidney disease is associated with cognitive impairment in studies of nondiabetic adults. We examined the cross-sectional relation between three measures of renal function and cognitive impairment (CI) in type 2 diabetic patients. A total of 357 patients with type 2 diabetes were prospectively enrolled. There were 108 patients with CI and 249 patients without CI (control). We calculated the urinary albumin/creatinine ratio (UACR) from morning spot urine and the estimated glomerular filtration rate (eGFR) in serum samples. Serum Cystatin C (Cys C) was measured with an automated particle-enhanced turbidimetric immunoassay. UACR and Cystatin C levels were significantly higher in patients with CI than those without CI (P<0.001), and the eGFR was lower in patients with CI than those without (P=0.003). A logistic regression analysis indicates that kidney impairment biomarkers levels were significantly associated with an increased risk of CI after adjustment for age and gender. The OR of each kidney biomarker (eGFR, UACR, Cystatin C) for CI status was 1.78 (0.89-3.27), 2.36 (1.29-4.42), and 2.77 (1.36-5.97), respectively. Among three kidney biomarkers (eGFR, UACR, Cystatin C), only elevated serum Cystatin C was associated with increased risk of CI in type 2 diabetic patients, with an OR of 1.42 (1.25-4.24) after additional adjustment for duration of diabetes, hypertension, hyperlipidemia, hemoglobin A1c (HbA1c), high-sensitivity C-reactive protein (Hs-CRP), intima-media thickness (IMT), ankle brachial index (ABI), and brachial-ankle pulse wave velocity (ba-PWV). Furthermore, combination of conventional risk factors and Cystatin C levels exhibited a fair diagnostic value for CI, with an area under the curve (AUC) of 0.91. Among three kidney impairment biomarkers (eGFR, UACR, Cystatin C), only elevated serum Cystatin C was associated with increased risk of CI in type 2 diabetic patients, independent of conventional risk factors. Furthermore, Cystatin C may be a better marker for CI than eGFR and UACR, and exhibited diagnostic value.

TRPM7 promotes the metastatic process in human nasopharyngeal carcinoma.

Our study observed the relationship between transient receptor potential melastatin 7 (TRPM7) expression and the metastatic process of nasopharyngeal carcinoma (NPC). We found that TRPM7 was overexpressed in 102 out of 206 (49.5%) human NPC cases and was significantly associated with clinical stage and lymphatic and distant metastasis. The results suggested that TRPM7 promotes NPC cell migration and invasion in vitro. Further, TRPM7 was correlated with poor clinical outcome and was an independent predictor for 5-year overall survival rate (HR, 1.832; 95% CI, 1.237-4.146 [P = 0.041]). In conclusion, TRPM7 promotes the metastasis of NPC and may serve as a prognostic marker in NPC patients.

Associations between biomarkers of renal function with cerebral microbleeds in hypertensive patients.

BACKGROUND: Cerebral microbleeds (CMBs) have been observed in the elderly and have been regarded as a manifestation of small vessel disease (SVD). Cerebral and glomerular SVD may have a common source of pathogenesis because these organs are closely connected through anatomic and hemodynamic similarities. The purpose of this study was to clarify the associations between kidney biomarker levels and CMBs in hypertensive patients. METHODS: The presence and number of CMBs were assessed on susceptibility-weighted imaging. We calculated the urinary albumin/creatinine ratio (UACR) from morning spot urine and the estimated glomerular filtration rate (eGFR) in serum samples. Serum cystatin C (CysC) was measured with an automated particle-enhanced turbidimetric immunoassay. RESULTS: UACR and CysC levels were higher in the patients with CMBs than those without, and the eGFR was lower in the patients with CMBs than those without. A logistic regression analysis indicates that eGFR and UACR were independently associated with the prevalence of deep or infratentorial CMBs. The odds ratio (OR) (95% confidence interval (CI)) of eGFR and UACR was 1.95 (1.37-3.27) and 2.25 (1.66-4.46), respectively. CysC was independently associated with CMBs in both deep or infratentorial and lobar locations. The ORs (95% CI) were 2.59 (1.57-6.22) and 1.57 (1.15-4.85), respectively. Furthermore, CysC exhibited fair diagnostic value for CMBs, with an area under the curve of 0.80. CONCLUSIONS: Kidney biomarker levels are associated with the presence of CMB in hypertensive patients without a history of transient ischemic attack (TIA) or stroke, independent of conventional risk factors, and CysC was a better marker for CMBs than eGFR and UACR.

Serum cystatin C and cerebral microbleeds in patients with acute cerebral stroke.

Recent studies have shown that kidney dysfunction is associated with cerebral microbleeds (CMB). Cystatin C is a more useful measurement than creatinine-based estimating equations for evaluating kidney function. The purpose of this study was to clarify the relationship between cystatin C levels and CMB in patients with acute cerebral stroke. This cross-sectional study included a total of 485 patients with acute ischemic stroke and 129 patients with cerebral hemorrhage. The serum levels of cystatin C were significantly higher in acute cerebral stroke patients with CMB than in those without (p<0.001). Multivariate logistic regression analyses showed that for each single standard deviation increase of cystatin C levels, there was a significant increase in the presence of CMB after adjusting for age and sex, and after additional adjustment for cardiovascular risk factors, silent lacunar infarction, and white matter hyperintensity in patients with acute stroke. The odds ratio (95% confidence interval) in patients with acute cerebral infarction and cerebral hemorrhage were 2.92 (1.81-6.93) and 2.98 (1.76-6.97), respectively. The present study suggests that elevated levels of cystatin C are associated with the presence of CMB in acute stroke patients, independent of conventional risk factors.

Immune mechanism and clinical significance of macrophage to medullary hematopoietic injury of immune-related hematocytopenia patients.

BACKGROUND: Immune-related hematocytopenia (IRH) is considered to be related with the production of autoantibody, as well as the activation of humoral immunity which is stimulated by B lymphocyte. This study aimed to observe the levels of various cytokines in the blood serum and the in situ active state of macrophage (Mφ) in the medullary hematopoietic microenvironment of IRH patients, and to probe into the immune mechanism and clinical significance of Mφ in hematopoietic cell injury. METHODS: ELISA is used to detect the IL-4, IL-6, IL-12, IL-17, and IFN-γ levels in the peripheral blood serum of 376 patients in pre- and post-therapy. Cytochemistry and cell immunochemistry methods are used to observe the peroxidase (POX), nonspecific esterase (NSE), hemosiderin granules, and HLA-DR activity of Mφ in the bone marrow of patients. Immunofluorescence is used to observe the expression of hemocyte antihuman globulin IgG antibody, lymphocytes CD4 molecule, Mφ membrane FcγIIreceptor (FcγIIR), mannitose receptor (MR), IFN-γ, ICAM-1, IL-12, and IL-17A and the formation mechanism of antibody-dependent cell-mediated cytotoxicity (ADCC) hematopoietic cell islands (HI) in the medullary hematopoietic microenvironment of patients. Glucocorticoid is used for treatment on the basis of anti-infection therapy, and gamma globulin stoss therapy is used for the appearance of ADCC-type HI or serious Mφ bloodthirsty phenomenon; if necessary, association of Cyclosporine A (CsA) should be used and chalybeate should be supplemented. RESULTS: In the patient group, the levels of IL-4, IL-6, IL-12, IL-17, and IFN-γ were increased. After treatment, the cytokine levels gradually became normal. The activated Mφ in the marrow highly expressed NSE and POX, and Mφ swallowed more hemosiderin particles, but the iron in the cytoplasm of immature erythrocytes decreased. The activated Mφ expressed HLA-DR, MR, ICAM-1, IFN-γ, and IL-12. For patients with humoral immunity activation and bacterial infection, Mφ weakly expressed IL-17A but highly expressed FcγIIR, and the phenomenon that ADCC-type HI broke pathological blood corpuscles often occurred; for the cellular immune activation along with virus infection, the white blood count (WBC) significantly reduced, Mφ weakly expressed FcγIIR, secretory highly expressed IL-17A, and the phenomena that Mφ adhered to, captured and swallowed blood cell often occurred. After four weeks of anti-infective and immunosuppressive therapy, nuclear apoptosis of Mφ occurred in the bone marrow of patients, HI and bloodthirsty phenomenon disappeared, and the peripheral blood picture started to improve. CONCLUSIONS: Mφ is an important antigen presenting cell in the IRH marrow for hematopoiesis destruction and an immune effector cell of hematopoietic injury; infection can promote the activation of Mφ, upregulate the impression of immune molecule and receptors, form ADCC HI, aggravate hematopoietic injury, and accelerate the destruction on hematopoietic cell.

[The immunological effect of anti-leukemic tumor induced by eosinophilic granulocyte].

OBJECTIVE: To investigate the biological effect of anti-leukemic cells induced by eosinophilic granulocyte (EOS) in bone marrow of patients with chronic myelogenous leukemia (CML). METHODS: The BCR-ABL fusion gene as well as the expression of IL-12 and IL-17 mRNA were performed by RT-PCR. The serum concentrations of cytokine IL-12 and IL-17 were determined by enzyme-linked immuno sorbent assay (ELISA). Immunochemistry staining and cytochemistry staining were used to observe the peroxidase (POX) and human leukocyte antigen (HLA)-DR expression of EOS in bone marrow. Immunofluorescence staining was used to observe mannose receptor (MR), IL-12, IL-17A and IL-17 receptor A (IL-17RA) expression of EOS. The results between the CML patients and the healthy controls were compared. RESULT: Serum levels of IL-12 and IL-17 were higher in the 60 CML patients [(196.33 ± 21.79) ng/L and (36.55 ± 3.01) ng/L] than those in the controls [(96.60 ± 4.92) ng/L and (23.74 ± 1.36) ng/L]. In the 32 patients with activated EOS, the levels of IL-12 and IL-17 were (273.12 ± 17.16) ng/L and (40.11 ± 6.13) ng/L, which were significantly higher than those in the non-activated EOS [(126.16 ± 14.27) ng/L and (28.14 ± 5.29) ng/L] (P values < 0.01). IL-12 and IL-17 mRNA were expressed in activated EOS, while BCR-ABL fusion gene was not found. The amounts of EOS were increased abnormally in the bone marrow and peripheral blood of the CML patients with POX positive staining in the cytoplasm and weakly positive HLA-DR staining. It was observed easily by a microscope that EOS could attack leukemic cells in bone marrow through adhesion, capture and phagocytosis. Activated EOS could express IL-12, IL-17A and MR, which was related with the serum levels of these cytokines. CONCLUSIONS: Activated EOS in bone marrow of CML patients could express IL-12 and IL-17. Activated EOS could induce coup injury to leukemic cell by releasing POX and expressing IL-12 and IL-17. It can also capture or swallow target cells via the expression of MR on the membrane. EOS may play an important role in the anti-tumor immunologic function in bone marrow of CML patients.

[Effects of cadmium stress on photosynthetic functions of strawberry].

With pot experiment, this paper studied the effects of cadmium stress on the leaf chlorophyll content, photosynthetic rate, stomatal conductance, transpiration rate, and intercellular CO2 concentration of strawberry. The results showed that cadmium reduced the chlorophyll content, and changed the chlorophyll a/b ratio. Cadmium reduced the photosynthetic rate and stomatal conductance, though they were increased by low concentration cadmium at the initial stage of cadmium stress. Cadmium also reduced the transpiration rate and intercellular CO2 concentration, but the decrement of intercellular CO2 concentration was relatively less.