OBJECTIVETo explore the effect of liposomal transfection of cyclin A antisense oligodeoxynucleotide (ASON) on HL-60 cell proliferation and apoptosis.

METHODSBy liposomal transfection, cyclin A ASON was co-cultured with HL-60 cells, the cell growth curve was determined by MTT assay and cell apoptosis electron-microscopy in situ cell apoptosis detection kit (POD), the protein and mRNA of cyclin A and bcl-2 were measured by FACS and RT-PCR, the role of cyclin A ASON in the development of leukemia was tested by the tumor formation in nude mice.

RESULTS(1) In the cyclin A ASON liposomal transfection group (group A), the proliferation of HL-60 cell was significantly inhibited as compared to those in cyclin A ASON group (group B) (68.9% vs 24.8%) (P < 0.01). (2) The expressions of cyclin A and bcl-2 of group A were significantly lower than those in the control group (1.1% vs 38.8%, P < 0.01; 21.9% vs 65.0%, P < 0.01, respectively), and the DNA ladder and apoptosis body was displayed. (3) In group A, the rate of tumor formation in nude mice was lower, the time for tumor formation was longer and the volume of tumor was smaller than those in control group.

CONCLUSIONLiposomal transfection of cyclin A ASON can inhibit in vitro proliferation of leukemia cells and induce in vivo apoptosis of the tumor cell, which might provide a new target for gene therapy.

Animals ; Apoptosis ; drug effects ; Cell Division ; drug effects ; Cyclin A ; genetics ; physiology ; Genetic Therapy ; HL-60 Cells ; Humans ; Leukemia ; therapy ; Liposomes ; Mice ; Mice, Inbred BALB C ; Oligonucleotides, Antisense ; pharmacology ; Transfection

OBJECTIVETo study the inhibition effect of cyclin G(1) antisense oligodeoxynucleotides (ASON) on the growth of HL-60 cells in nude mice.

METHODS(1) Nude mice were divided into control group, sense oligodeoxynucleotides (SON) group and ASON group. After (60)Co radiation, with HL-60 cells SON group and ASON group were subcutaneously innoculated; (2) The weight and volume of tumors were continually measured; (3) The morphology of tumor cells was observed by microscope; (4) The protein and mRNA expression levels of cyclin G(1) were determined by flow cytometry (FCM) and reverse transcription polymerase chain reaction (RT-PCR); (5) The cell apoptosis was detected by electron microscopy and FCM.

RESULTS(1) The inhibition rate of tumor in ASON group was 69.4%. In ASON group, the wight and volume of tumor were significantly lower than those in SON group and control group. (2) The HL-60 cells in ASON group showed morphologically smaller nuclei, less mitosis, less heteromorphosis and apoptosis.

CONCLUSIONThe cyclin G(1) ASON can inhibit the growth of HL-60 cells in nude mice and induce apoptosis.

Animals ; Apoptosis ; genetics ; Cell Division ; genetics ; Cyclin G ; Cyclin G1 ; Cyclins ; genetics ; metabolism ; Female ; Flow Cytometry ; HL-60 Cells ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Oligonucleotides ; genetics ; metabolism ; Oligonucleotides, Antisense ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Xenograft Model Antitumor Assays ; methods

OBJECTIVETo analyze the association between G894T (Glu298Asp) mutation at exon 7 in the endothelial nitric oxide synthase gene and essential hypertension.

METHODSOne hundred and sixteen essential hypertensives without taking hypertensive medication and 136 normotensives screened from health workers in a steel factory were selected as subjects in this study. Polymerase chain reaction (PCR) and Ban II restriction enzyme digestion were performed to detect the G894T mutation.

RESULTSG894T mutation was significantly associated with essential hypertension. The T allele frequency in essential hypertensive group was significantly higher than that in normotensive group (16.0% versus 8.8%, P = 0.019, OR = 1.96, 95% CI: 1.14 - 3.37). The levels of systolic blood pressure and diastolic blood pressure in the G894T mutant genotypes were all significantly elevated in hypertensive, normotensive, and the total subjects (P < 0.05). After adjusting factors as age, sex, smoking, alcohol drinking, body mass index, triglyceride, serum total cholesterol, serum high density lipoprotein cholesterol by analysis of multiple covariance, significant positive effect of the G894T mutant genotypes on blood pressure in the total subjects (P < 0.01) was noticed.

CONCLUSIONThis study suggested that the G894T mutation in the endothelial nitric oxide synthase gene might serve as a major risk factor of essential hypertension in this study population.

Adult ; Alleles ; Blood Pressure ; genetics ; China ; epidemiology ; Exons ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Hypertension ; enzymology ; genetics ; Male ; Middle Aged ; Nitric Oxide Synthase ; genetics ; Nitric Oxide Synthase Type III ; Point Mutation ; Polymorphism, Restriction Fragment Length ; Risk Factors

OBJECTIVETo explore the relationship between the 7th exon G894T mutation of endothelial nitric oxide synthase (eNOS) gene and overweight in patients with essential hypertension.

METHODSTotally, 116 patients with essential hypertension taking no medications and 136 normotensives were selected from a steel workers as study subjects. Polymerase chain reaction and restriction fragment length polymorphism were performed to detect mutation of the 7th exon G894T. Additive model was used to analyze interaction between G894T mutation and overweight on hypertension. Population attributable risk percent (PAR%) for them, etiologic fraction, was applied to their contribution to hypertension.

RESULTSThere was a positive interaction between G894T mutation and overweight on essential hypertension, with an index of interaction of 1.99 and attributable interaction percent of 30.76%. Their pure attributable interaction percent was 36.38%. Multiple logistic regression analysis showed that there still was positive interaction between G894T mutation and overweight on essential hypertension, adjusted for age, sex, smoking and alcohol drinking. Index of their attributable interaction was 2.85, with attributable interaction percent of 39.97%, also adjusted for the above-mentioned factors. Their pure attributable interaction percent was 46.49% and PAR% was estimated as about 15% under certain condition.

CONCLUSIONSInteraction between mutation of the 7th exon G894T of eNOS gene and overweight played an important role in essential hypertension of the studied population. Control of body weight in the population with both G894T mutation and overweight could markedly decrease their risk of hypertension.

Adult ; Exons ; Female ; Humans ; Hypertension ; etiology ; Male ; Middle Aged ; Mutation ; Nitric Oxide Synthase ; genetics ; Nitric Oxide Synthase Type III ; Obesity ; complications

OBJECTIVEThe purpose of our study is to observe the voltage-gated potassium channel Kv1.3 expression on CD4+CD28null T cells from the peripheral blood of ACS patients by the patch clamp technique.

METHODSKv1.3 potassium channels expression from 17 patients with ACS and 11 healthy age-matched normal controls was detected in single cell (CD4+CD28null T cells and CD4+CD28+ T cells) by fluorescence microscopy and patch clamp.

RESULTSThe percent of CD4+CD28nullT cells are higher in the ACS (6.97% +/- 2.05%) than that in the controls (1.38% +/- 0.84%, P < 0.05). The concentration of hsCRP is directly correlated with the number of the CD4+CD28null T cells in the ACS (r = 0.52, P < 0.05). The conductance [(6.89 +/- 1.17) nS vs. (3.36 +/- 0.66) nS], dens [(1.95 +/- 0.80) n/microm2 vs. (1.13 +/- 0.57) n/microm2] and numbers [(574.5 +/- 97.6) n/cell vs. (280.3 +/- 55.3) n/cell] of the Kv1.3 channels on the CD4+CD28null T cells are significantly higher than those on the CD4+CD28+ T cells (all P < 0.01) in ACS patients, but were similar on CD4+CD28+ T cells between ACS patients and controls.

CONCLUSIONThe CD4+CD28null T cells in the ACS and the numbers of Kv1.3 channels on the CD4+CD28null T cells from the ACS patients are significantly upregulated and might contribute to the pathogenesis of ACS.

Acute Coronary Syndrome ; blood ; immunology ; metabolism ; Aged ; CD28 Antigens ; metabolism ; CD4-Positive T-Lymphocytes ; metabolism ; Case-Control Studies ; Female ; Humans ; Kv1.3 Potassium Channel ; metabolism ; Male ; Middle Aged ; Patch-Clamp Techniques

Objective To assess the dietary risk of organophosphates pesticides residues in some vegetables in Shaanxi Province. Methods The monitoring data of organophosphorus pesticides in vegetables from 2012 to 2016 were derived from Shaanxi Province Food Contamination Monitoring Network. The exposure of organophosphorus to vegetables by using exposure risk index (ERI) , dietary exposure risk index (RI) and dietary exposure risk assessment. Results Exposed risk index (ERI) of organophosphorus in some vegetables in Shaanxi Province ranged from 1.25E-06 to 1.87E-01, and among them, the highest ERI of clozaptid in garlic was 1.87E-01, followed by onion. In some vegetables, the highest exposure to organophosphorus was isocarbophos of fresh beans with an exposure of 4.27E-02 μg / (kg body weight · day) and the lowest dietary exposure was bulbs, and stems and fresh beans were 2.00E-04μg / (kg body weight·day) . The exposure of organophosphorus to all kinds of vegetables was less than their respective daily allowable intake (ADI) . The risk of dietary exposure to organic- phosphorus in vegetables was 187％ and greater than 100％ for the risk index (RI) of lettuce, and RI for all other vegetables was <100％. Conclusion The dietary exposure and risk index of organophosphates pesticides in some vegetables in Shaanxi Province was safe, and more attention should be paid to dimethoate and isazofos.

OBJECTIVE: To investigate the protective effects and its possible mechanism of Wuzi Yanzong Pill (WYP) on Parkinson's disease (PD) model mice. METHODS: Thirty-six C57BL/6 male mice were randomly assigned to 3 groups including normal, PD, and PD+WYP groups, 12 mice in each group. One week of intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was used to establish the classical PD model in mice. Meanwhile, mice in the PD+WYP group were administrated with 16 g/kg WYP, twice daily by gavage. After 14 days of administration, gait test, open field test and pole test were measured to evaluate the movement function. Tyrosine hydroxylase (TH) neurons in substantia nigra of midbrain and binding immunoglobulin heavy chain protein (GRP78) in striatum and cortex were observed by immunohistochemistry. The levels of TH, GRP78, p-PERK, p-eIF2α, ATF4, p-IRE1α, XBP1, ATF6, CHOP, ASK1, p-JNK, Caspase-12, -9 and -3 in brain were detected by Western blot. RESULTS: Compared with the PD group, WYP treatment ameliorated gait balance ability in PD mice (P<0.05). Similarly, WYP increased the total distance and average speed (P<0.05 or P<0.01), reduced rest time and pole time (P<0.05). Moreover, WYP significantly increased TH positive cells (P<0.01). Immunofluorescence showed WYP attenuated the levels of GRP78 in striatum and cortex. Meanwhile, WYP treatment significantly decreased the protein expressions of GRP78, p-PERK, p-eIF2α, ATF4, p-IRE1 α, XBP1, CHOP, Caspase-12 and Caspase-9 (P<0.05 or P<0.01). CONCLUSIONS WYP ameliorated motor symptoms and pathological lesion of PD mice, which may be related to the regulation of unfolded protein response-mediated signaling pathway and inhibiting the endoplasmic reticulum stress-mediated neuronal apoptosis pathway.