查看更多>>摘要：Objective:Gold nanoparticle Hepatitis B virus (HBV) DNA probes were prepared, and their application for HBV DNA measurement was studied. Methods: Alkanethiol modified oligonucleotide was bound with self-made Au nanoparticles to form nanoparticle HBV DNA gene probes, through covalent binding of Au-S. By using a fluorescence-based method, the number of thiol-derivatized, single-stranded oligonucleotides and their hybridization efficiency with complementary oligonucleotudes in solution was determined. With the aid of Au nanoparticle-supported mercapto-modified oligonucleotides serving as detection probes, and oligonucleotides immobilized on a nylon membrane surface acting as capturing probes,HBV DNA was detected visually by sandwich hybridization based on highly sensitive aggregation and silver staining. The modified nanoparticle HBV DNA gene probes were also used to detect the HBV DNA extracted from serum in patients with hepatitis B. Results: Compared with bare Au nanoparticles, oligonucleotide modified nanoparticles had a higher stability in NaCl solution or under high temperature environment and the absorbance peak of modified Au nanoparticles shifted from 520nm to 524nm. For Au nanoparticles, the maximal oligonucleotide surface coverage of hexaethiol 30-mer oligonucleotide was (132 ± 10) oligonucleotides per nanoparticle, and the percentage of hybridization strands on nanoparticles was (22 ± 3%). Based on a two-probe sandwich hybridization/nanoparticle amplification/silver staining enhancement method, Au nanoparticle gene probes could detect as low as 10~(-1) mol/L composite HBV DNA molecules on a nylon membrane and the PCR products of HBV DNA visually. As made evident by transmission electron microscopy , the nanoparticles assembled into large network aggregates when nanoparticle HBV DNA gene probes were applied to detect HBV DNA molecules in liquid. Conclusion:Our results showed that successfully prepared Au nanoparticle HBV DNA gene probes could be used to detect HBV DNA directly. The detection-visuallized method has many advantages, including high sensitivity, simple operation and low cost This technique has potential applications in many fields, especially in multi-gene detection chips.
查看更多>>摘要：Objective:To evaluate Tpeak-Tend (Tp-e) interval in surface standard ECG as a new risk factor for arrhythmic event in patient with Brugada syndrome. Methods: 23 male patients with Brugada syndrome and 20 male patients with paroxysmal supraventricular tachycardia (PSVT) as the control group were investigated in this study. Tp-e interval in surface standard ECG was compared between BrS and PSVT patients. Results: Tp-e interval in BrS patients was significantly longer than that in PSVT patients (109.57±22.86 ms vs. 88.50±13.08ms, P< 0.05). There was significant difference in Tp-e interval between 16 BrS patients with arrhythmic events (including syncope, clinical ventricular fibrillation [VF] and programmed electrical stimulation [PES]-induced VF) and 7 BrS patients without arrhythmic events and PSVT patients (118.12±20.40ms vs. 90.00±l5.27ms, P< 0.05; 118.12±20.40ms vs. 88.50±13.08ms, P< 0.05). However, Tp-e interval was similar in BrS patients without arrhythmic events and PSVT patients (90.00±15.27ms vs 88.50±13.08ms, P>0.05). Conclusion; The prolongation of Tp-e interval could serve as a new noninvasive event predictor for arrhythmic events in patients with Brugada syndrome.
查看更多>>摘要：Objective: To study the gene mutations of homeobox transcription factor (CSX/NKX_(2.5)) associated with a Chinese family with secundum atrial septal defect (ASD). Methods: Polymerase chain reaction and DNA sequencing were used to check all the members in the family with ASD, and single strand conformation polymorphism analysis (SSCP) was used to check 126 normal control people for detecting the mutations of CSX/NKX_(2.5) gene. Results: Three mutations,G270A(Glu32Lys) ,G378A (Glu68Lys)andG390A (Glu72Lys)were identified in CSX/NKX_(2.5) gene of ASD patients. However, the other members in the family with ASD and the control did not have such gene mutations. Conclusion:These mutations of CSX/NKX_(2.5) gene, which were identified in a Chinese family, may be one of the secundum ASD etiologic causes.
查看更多>>摘要：Objective:To evaluate the association of the peripheral levels of the defined inflammatory markers with different types of acute coronary syndrome (ACS) and stable angina, and the role inflammation played in the pathogenesis of ACS. Methods:For understanding the variation of serum concentrations of matrix metalloproteinase-9 (MMP-9), soluble intercellular adhesion molecule-1 (sICAM-l),C-reactive protein(CRP), and white blood cell count(WBC) and their association with ACS, 90 patients with coronary heart disease (CHD) and 30 healthy volunteers were recruited. The enrolled people were assigned into four equal groups, including acute myocardial infarction (AMI) group, unstable angina pectoris(UAP) group, stable angina pectoris(SAP) group and healthy control group. The serum levels of MMP-9 and sICAM-1 were measured with ELISA kits, CRP were measured with im munoturbidimetric assay, and WBC number were assessed all before any treatment was administrated. Results: (1)The serum levels of MMP-9,sICAM-1,CRP and WBC in the patients with ACS were significantly higher than those in the control group (P < 0.01). (2)Compared with control group, patients with SAP only had higher serum level of sICAM-1 (P < 0.01). While the levels of MMP-9, CRP, and WBC had no significant difference between them (P > 0.05 all). (3)Significant positive correlation between the serum levels of MMP-9 and sICAM-1 and CRP and WBC all were observed in the patients with ACS (P < 0.05). Conclusion: The elevation of serum concentrations of inflammatory markers including MMP-9, sICAM-1, CRP and WBC were associated with initiation and progression of ACS, and they may help predicting cardiovascular events.
查看更多>>摘要：Objective: To evaluate the effect of osteogenic potential on human marrow mesenchymal stem cells (hMSCs) transferred with human vascular endothelial growth factor (VEGF) gene by adenovirus. Methods: hMSCs were isolated from human marrow, cultured in vitro and randomly divided into 3 groups:Ad-VEGF_(165) group: adding adding 1×10~(10)OPU/ml Ad -VEGF in hMSCs culture fluid after incubating 24 hours, changing into ordinary complete culture and continuing culturing; Positive control group: Cultured hMSCs with 1 nmol/L dexamethasone, 10 mmol/L glycerophosphate and 50 mg/L vitamin C, exchanging this conditioned medium twice a week; blank control group:no special treatment but culturing hMSCs in DMEM.To evaluate osteogenesis competence, Von Kossa's staining and a quantitative alkaline phosphates (ALP) activity analysis were performed after 2 weeks treatment. Results: The calcified nodes formed after 2 weeks treatment in Ad-VEGF165 group and Positive control group but not in blank control group. ALP activities in Ad-VEGF165 group, Positive control group and blank control group were (7.91 ± 0.90)u/L, (8.18 ± 0.76 u/L) and (3.46 ± 0.49)u/L respectively. The differences were no statistical significance between Ad-VEGF165 group and positive control group (P > 0.05), but Ad-VEGF165 group and Positive control group were significantly different with blank control group (P < 0.05). Conclusion: Adenovirus mediated VEGF165 gene can transfect hMSCs and promote osteogenesis of hMSCs.
查看更多>>摘要：Objective:To observe the tolerance and the dependence of endomorphin-1 (EM-1) in rats and the possible mechanisms. Methods:Sixty Sprague-Dawley rats were randomly allocated into saline, acute EM-1-treated and chronic EM-1-treated groups. The rats were intracerebroventricularly injected with saline, acute EM-1 10 μg/kg 30 min prior to sacrifice,and chronic EM-1 by daily administration at 8:00 A.M. and 15:00 P.M. from 10 μg/kg on the 1st day to 50 μg/kg on the 9th day, respectively. In chronic EM-1-treated group, the median antinociceptive dose (AD_(50)) and the catatonic median effective dose (ED_(50)) were determined by the improved Dixon's method. Natural withdrawl test was used to assess the dependence of EM-1. Maximal binding capacity (Bmax) and dissociation constant(Kd) of ~3H-DAMGO, binding to mu-opioid receptor (MOR) in brain tissue, was measured by Scatchard analysis. Gene expression of MOR was measured by reverse transcription-polymerase chain reaction (RT-PCR). Results:Tolerance of the antinociceptic and catatonic effects on the 3rd day(3.1-fold and 1.9-fold) and the 9th day(28.4-fold and 8.5-fold) were observed in chronic EM-1-treated group (P < 0.05). Jumping times and withdrawal scores of rats were significantly higher in the chronic EM-1-treated group than those in saline group on the 9th day (P < 0.05). Bmax and mRNA expression of MOR in cortex, midbrain and striatum were lower in chronic EM-1-treated group on the 9th day than the other two groups(P < 0.05), but Kd had no significant difference (P > 0.05). AD_(50),ED_(50), Bmax,Kd and gene expression of MOR were recorded. Conclusion: EM-1 possesses the tolerance and the dependence. After a long-term treatment, EM-1 down regulates the binding capacity and mRNA of MOR, which somewhat accounts for the dependence.
查看更多>>摘要：Objective:To investigate the apoptosis of dopaminergic neurons and the protective effect of nicotine in 1-methyl-4-pheny 1-1, 2,3,6-tetrahydropyridine (MPTP)-induced mouse model of Parkinson's disease. Methods:The mouse model of Parkinson's disease were formed by MPTP (30 mg/kg/d×7, i.p.); and the loss and apoptosis of dopaminergic neurons was observed by Tyrosine Hydroxylase(TH) and TUNEL stains.In "Nicotime plus MPTP" group, mice were pretreated with nicotine before MPTP injection. The putative protective effect of nicotine was analyzed. Results:The number of TH-positive cells decreased during MPTP treatment. Apoptotic neurons began to appear after three injections of MPTP and peaked on the 8th day. In the MPTP-intoxicated mice treated with nicotine, the loss of TH-positive cells was significantly less than that of MPTP-treated group (30 mg/kg/d×7) (P < 0.05). Conclusion:The chronic treatment of MPTP can induce the apoptosis of dopaminergic neurons in substantia nigra, and nicotine might have a neuroprotecitve effect on dopaminergic neurons against MPTP toxicity.
查看更多>>摘要：Objective:To evaluate the effect of HIF-1α over-expression on angiogenesis in human prostate cancer cells. Methods: LNCaP cells(a human prostate cancer cell line) were transfected with the recombinant plasmid pcDNA3.1(-)-HIF-1α with Lipo-fectamine 2000 system. The positive clones were selected by G418 being further confirmed by Western blot and immunofluores-cence. The expression levels of VEGF, iNOS and Ang-E were determined. Results:The expression of HIF-1α in the LNCaP/ HIF-1α cells was significantly increased in transfected cells, which induced the up-regulation of VEGF, iNOS, whereas Ang-Ⅱ expression remained un- changed. Conclusion:Over-expression of HIF-1α can induce angiogenesis proteins and may improve the angiogenesis potency of prostate cancer.
查看更多>>摘要：Objective:To investigate glucose metabolism in women with multiple ovarian follicles (MOF) and explore the relationship between glucose metabolism, insulin resistance and body weight. Methods:We evaluated 46 women with MFO and 30 normal women as controls. All the subjects were given 75g of glucose orally in order to perform the oral glucose tolerance test (OGTT) and insulin releasing test(IRT), and they were also evaluated for insulin resistance using the insulin resistance index with homeostatic model assessment (HOMA). Results: The occurrence of impaired glucose tolerance in women with MOF was 10.87%, which was significantly higher than that in the control group (3.33%,P < 0.05). The rate of insulin resistance was 30.43% in the study group as compared to 10.00% in the control group. The results showed that there was significant difference between the two groups(P < 0.05). The levels of FSH,LH,PRL,E_2,T and P between the two groups had no significant difference (P > 0.05). BMI in women with impaired glucose tolerance was correlated positively to insulin resistance (r = 0.567, P < 0.05). Conclusion:Abnormal glucose metabolism was observed in women with unitary multiple ovarian follicles, and this could be attributed to obesity and insulin resistance. Women with MOF and associated obesity should be subjected to OGTT so that their glucose levels can be monitored as a preventive measure.
NETL
NSTL