Determination of salsolinol N-methyltransferase activity in 6-OHDA-lesioned rats / 中国比较医学杂志

Objective The aim of this study was to establish a rat model of Parkinson''s disease ( PD) by using 6-hydroxydopamine (6-OHDA) and detect the salsolinol N-methyltransferase ( SNMT) activity in peripheral lymphocytes of PD rats for the development of a biomarker for early diagnosis of PD. Methods Rat model of PD was established by unilateral double-pointed injection of 6-OHDA into the striatum and was verified by behavior observation. An analytical method was developed based on multiple reaction monitoring with HPLC-ESI-QQQ to determine the SNMT activity in peripheral lymphocytes. Results Seven of 18 rats injected with 6-OHDA showed steadily apomorphine-induced rotation ( >7 r/min) . The success rate was 38. 9%. A sensitive and stable quantitative method with internal standard added was created, based on multiple reaction monitoring mode to analyze SNMT activity. The limit of detection ( LOD) and limit of quantitation ( LQD) of N-methyl-salsolinol, which is the product of Salsolinol catalyzed by SNMT, were 49 pmol/L and 98 pmol/L, respectively. The precisions of intra-day and inter-day assays both were below 6. 0%. SNMT activity of peripheral lymphocytes in the 6-OHDA-lesioned rats was significantly increased [43. 37 ±9. 49 pmol/(h·mg)NMSal] in comparison with that in the normal group [2. 16 ±5. 82 pmol/(h·mg)NMSal] and the sham-operated group [0. 58 ±2. 32 pmol/(h· mg)NMSal](P< 0. 01, n=5). There was no significant difference between the normal group and sham-operated group (P< 0. 05, n =5). Conclusions Our results indicate that SNMT activity may reflect the changes in the course of Parkison''s disease and may become a potential clinical biomarker in diagnosis of this disease.

Altered Cell Viability and Proliferation Activity of Peripheral Lymphocytes in Patients with Alzheimer's Disease

OBJECTIVE: We evaluated cell viability and proliferation activity of peripheral lymphocytes as potential models of neuronal death in Alzheimer's disease (AD). METHODS: We analyzed the cell viability and proliferation activity of phytohemagglutinin (PHA)-activated lymphocytes from 68 AD patients and 33 normal controls. The cellular measures were made at baseline (0 hr), 24 hrs, 48 hrs, 72 hrs, and 96 hrs after PHA stimulation. RESULTS: Cell viability in the AD patients was significantly decreased at 72 hrs and 96 hrs, compared with the normal controls. The declining ramp of the proliferation activity from 48 hrs to 72 hrs after PHA stimulation was significantly related to cell viability at 72 hrs and at 96 hrs in the AD patients. CONCLUSION: Lymphocytes from patients with AD have altered viability and proliferation characteristics in culture following PHA stimulation. These findings suggest that lymphocytes may be used as a peripheral tissue model of cell cycle dysregulation in AD.

Effects of light on the expression patterns of circadian clock-related genes in the peripheral lymphocytes / 中华物理医学与康复杂志

Objective To explore the effects of light on the expression patterns of clock and clock-related genes in peripheral lymphocytes.To develop basic knowledge needed for clinical application of the circadian clock.Methods One hundred Sprague-Dawley rats were housed under constant dark(DD)or normal light-dark(LD 12:12)conditions for six weeks.Peripheral iymphoeytes were collected at different time points.The expression level of the clock gene and melatonin receptor genes mt1 and mt2 were detected using semi-quantitative RT-PCR.The data were analyzed with cosine software.Results Circadian expression of the genes was observed in both groups,but the peak phase,amplitude and strength of expression of each gene differed with the light conditions.Conclusion Light influences the expression of clock and clock-related genes in rats' peripheral lymphocytes.The clock gene might play an important role in regulating the expression of mt1 and mt2.

Cell Viability and Proliferation Activity of Peripheral Lymphocytes in Patients with Alzheimer's Disease / 신경정신의학

OBJECTIVES: There are evidences of apoptotic neuronal cell death in Alzheimer's disease (AD). Recent studies suggested AD pathogenesis in the central nervous system as well as in peripheral lymphocytes. The object of this study is to compare the cell viability and the proliferation activity in AD patients with healthy normal control by using peripheral lymphocytes. METHODS: We analyzed the cell viability and the proliferation activity of phytohemagglutinin (PHA)-activated lymphocytes from 73 AD patients and 31 normal contols. The cell viability and the proliferation activity were measured at baseline (T0), 24 hours (T24), 48 hours (T48), 72 hours (T72), 96 hours (T96), by the tryphan blue method and the BrdU proliferation activity method, respectively. RESULTS: The cell viability of PHA-activated peripheral lymphocytes in AD patients was significantly decreased at T72, T96 compared with healthy controls (F=8.034, p<0.001). In AD patients, the decline of proliferation activity appeared in earlier than healthy normal controls. CONCLUSION: This study suggests that there is a decreased cell viability and the proliferation activity of peripheral lymphocytes in AD patients. These finding may be related with the increased apoptosis in Alzheimer's disease.

A Study of Production of Interleukin-1alpha by Peripheral Lymphocytes in Patients with Coronary Artery Disease

BACKGROUND: Interleukin-1alpha is interesting lymphokine to cardiologists because it has been implicated as a regulatory protein in the development and clinical sequale of atherosclerosis, including the modulation of low density lipoprotein metabolism, the regulation of vascular smooth muscle cell mitogenesis, the stimulation of leukocyte adherence to endothelium, and procoagulant activity. But most interleukin-1alpha remains in the cytosol of cells in its precursor form, and is transported to cell surface. and associated with cell membrane. Therefore considerable amount of interleukin-1alpha, contrast to interleukin-1beta, is not released by cells into the extracellular space and the circulation. Despite of increased production of interleukin-1alpha, circulating level may not be elevated because of autocrine and paracrine action of that. In order to elucidate whether interleukin-1alpha is systematically elevated or not in patients with coronary artery disease who are complaining of chest pain, we undertook this study. METHODS: We isolated lymphocytes from peripheral blood in patients and control group. After the peripheral lymphocytes were cultured in the presence or absence of phytohemmagglutinin in RPMI-1640 media for 24 hours, we measured the content of interleukin-1alpha in supernatant by radioimmunoassay. RESULTS: 1) In the absence of phytohemagglutinin, the mean value of Interleukin-1alpha in the supernatant was 29.13+/-17.42 pmol/ml in control group and 27.28+/-18.80 pmol/ml in patients group(p=NS). 2) In the presence of phytohemagglutinin, the mean value of Interleukin-1alpha in the supernantant was 36.53+/-20.72 pmol/ml in control group and 152.13+/-91.85 pmol/ml in patient group(p<0.0001). CONCLUSION: Significant increase of interleukin-1alpha in the presence of phytohemagglutinin in the patient group means that the peripheral lymphocytes in patients with coronary artery disease are activated to produce interleukin-1alpha.