Preparation of Protein Hybrid Fluorescence Nanoclusters for Rapid Detection of Mercury Ion / 分析化学

One-step green synthetic approach,with bovine serum albumin(BSA) as stabilizer and reductant, was developed for preparation of BSA hybrid fluorescence gold nanoclusters (AuNCs@BSA). The prepared AuNCs@BSA exhibited strong red fluorescence under UV light illumination. Upon excited at 360 nm, the fluorescence spectrum of AuNCs@ BSA exhibited maximum emission peak at 635 nm. AuNCs@ BSA was presented as uniform spherical morphology with diameter at (2.0 ±0.05) nm. The fluorescence of AuNCs@BSA could be quenched by Hg2+because of its metallophilic reaction. Based on the fluorescent spectrometry, a rapid detection system was developed for Hg2+detection in tap water. The AuNCs@BSA amount, pH and buffer system were optimized in this study. According to optimization results, ultrapure water (pH 5.0) was selected to dilute the AuNCs@BSA by 100 times, and 50 μL/well of AuNCs@BSA dilution was applied to detect mercury ion in tap water. Under the optimized conditions, the detection could be completed within 3 min,the fluorescence intensity of the system was linearly proportional to the concentration of mercury ion in the range of 0.5–900 μg/L with linear equations y=-26.76lgx+803.1(0.5-75 μg/L,R2=0.9951) and y=-0.27x+762.02 (75-900 μg/L,R2=0.9959). The limit of detection was 0.14 μg/L(3σ). The average recoveries in spiked tape water samples ranged from 86.8%-113.4% with relative standard deviation of less than 15%. The result implied that the developed method was able to apply to detect mercury ion rapidly, sensitively and conveniently.

Attenuation and mechanism of endoplasmic reticulum stress-mediated hepatocyte apoptosis in rats with alcohol-induced liver injury by qinggan huoxue recipe and its disassembled formulas / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore attenuation and mechanism of endoplasmic reticulum stress (ERS)-mediated hepatocyte apoptosis in rats with alcohol-induced liver injury by Qinggan Huoxue Recipe (QGHXR) and its disassembled formulas (Qinggan Recipe and Huoxue Recipe respectively).</p><p><b>METHODS</b>A rat model of chronic alcoholic liver injury was successfully established using a compound reagent of alcohol, corn oil, and pyrazol. The modeled rats were randomly divided into the model group, the QGHXR group, the Qinggan Recipe (QGR) group, and the Huoxue Recipe group (HXR). The CCl4 control group and the normal control group were also set up. There were ten rats in each group. All rats of modeled groups were gastrogavaged with alcohol compound reagent every morning. Rats in the QGHXR group (at the daily dose of 9. 5 g/kg, QGR group (at the daily dose of 3.0 g/kg), and HXR group (at the daily dose of 6.5 g/kg) were administered with corresponding medicines by gastrogavage every afternoon. Equal volume of normal saline was given to rats of the model group by gastrogavage. CCl4 was intraperitoneally injected at the dose of 0.3 mL/kg to rats in the CCl4 control group, once per week. Normal saline was given to rats in the normal control group by gastrogavage. The treatment was lasted for two weeks. Pathological changes of the liver were observed by histopathology. Serum total homocysteine (tHCY) level was detected by ELISA. The hepatocyte apoptosis rate was detected using flow cytometry. The gene and protein expressions of eukaryotic translation initiation factor 2 alpha (elF-2alpha), phosphorylation elF-2alpha (pelF-2alpha), glucose-regulated protein 78 (GRP78), and Caspase-3 in the liver were examined using Real-time PCR and Westen blot respectively.</p><p><b>RESULTS</b>Compared with the normal control group, typical pathological changes of chronic alcoholic liver injury such as steatosis, inflammation, and even fibrosis occurred in model rats. The hepatocyte apoptosis obviously increased, with the apoptosis rate reaching the five-fold of that in normal rats. Besides, early apoptosis dominated. The serum tHCY level significantly increased. The expressions of p-elF-2alpha, GRP78, and Caspase-3 protein obviously increased (P < 0.01). Expressions of GRP78 and Caspase-3 mRNA significantly increased (P < 0.05, P < 0.01). Compared with the model group, the degrees of the liver injury and the hepatocyte apoptosis in the QGHXR group, the QGR group, and the HXR group were significantly alleviated. The serum tHCY level was significantly lowered. The protein expressions of p-elF-2a, GRP78, and Caspase-3 obviously decreased (P < 0.01). mRNA expressions of GRP78 and Caspase-3 obviously decreased in the QGHXR group (P < 0.05, P < 0.01). Only GRP78 mRNA expression obviously decreased in the QGR group (P < 0.05).</p><p><b>CONCLUSION</b>QGHXR and its disassembled formulas could attenuate ERS-mediated hepatocyte apoptosis in alcohol-induced liver injury rats by lowering the serum tHCY level and expressions of ERS apoptosis correlated factors.</p>

Role of PERK/eIF2a signaling pathway in hepatocyte apoptosis of alcoholic liver injury rats / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the role of PERK/eIF2alpha signaling pathway in hepatocyte apoptosis of alcoholic liver injury rats.</p><p><b>METHODS</b>Rat models with ethanol-induced liver injury were successfully developed by gastric gavage with ethanol-corn oil mixtures for 12 weeks. At different time points (4, 6, 10, 12 week), liver pathology was dynamically observed. The hepatocyte apoptosis was quantitatively analyzed by Annexin V-FITC/PI double-labeled flow cytometry, the serum total homocysteine (tHCY) level was detected by ELISA and the expressions of eIF2a, p-eIF2a, GRP78/Bip, GRP94, caspase-3 and caspase-12 in liver were examined using Real-time PCR and Western blot.</p><p><b>RESULTS</b>Typical acute liver injury and chronic liver injury were observed at week 4 and week 12 respectively. The hepatocyte apoptosis rates in 6-week model rats significantly increased compared with normal rats (P value less than 0.05), and the degree of hepatocyte apoptosis continued to increase with the modeling time, and the percentages of early and total apoptosis reached 26% and 29% at week 12. From week 6 to week 12, the serum tHCY levels in model rats were obviously higher than in normal rats (P value less than 0.01). Since week 4, eIF2a protein phosphorylation in model rat livers remarkably elevated compared with that in normal rat livers (P value less than 0.01), and at week 12 the peIF2a protein expression in model rat livers increased by 2.81-fold. Since week 4 the expressions of GRP78/Bip, GRP94, caspase-12 and caspase-3 mRNA and protein in model rat livers showed a significant increase as compared to normal rat livers, and at week 12, these gene and protein levels increased 4.70, 12.95, 3.83, 4.05 fold and 3.93, 6.93, 9.88, 3.31 fold, respectively (P value less than 0.01).</p><p><b>CONCLUSION</b>Activation of PERK/eIF2a signaling pathway contributes to the occurrence and development of hepatocyte apoptosis in alcoholic liver injury rats and it might be as a potential target for therapeutic applications in alcoholic liver diseases.</p>

Association between the polymorphism of GNB3C825T gene and vasovagal syncope in children / 中华儿科杂志

<p><b>OBJECTIVE</b>Investigate the association between GNB3C825T gene polymorphism and pediatric vasovagal syncope.</p><p><b>METHOD</b>Syncope group consisted of 54 cases of unexplained syncope in children, including 18 males and 36 females, at the age of 11.8 years; control group consisted of 54 healthy children over the same period, of whom 20 were male and 34 female, at the age of 11.2 years. The patients underwent head-up tilt test (HUTT). According to HUTT test results, HUTT-positive group and HUTT-negative group were further classified. For cases in HUTT-positive group, based on the changes in blood pressure and in heart rate during HUTT, vasodepressor, mixed and cardioinhibitory patterns were studied. DNA was extracted from peripheral blood in all the patients. A pair of primers was designed flanking 825 polymorphic loci. Products were recovered by using polymerase chain reaction (PCR). GNB3C825T polymorphism was detected by using gene-side GNB3C825T sequencing. Allele distribution between the various groups were studied.</p><p><b>RESULT</b>Among fifty-four children with syncope, HUTT was positive in 30 cases, including vasodepressor pattern in 15 cases (50.0%), mixed pattern in 9 cases (30.0%) and cardioinhibitory pattern in 6 cases (20.0%). Whereas the subjects in control group had negative HUTT response. GNB3C825T allele C in the control and syncope groups was 81.5% and 65.7%, respectively. GNB3C825T allele T in the control and syncope groups was 18.5% and 34.3%, respectively (χ(2) = 6.888, P < 0.05). GNB3C825T allele C in HUTT-positive and negative groups was 61.7% and 81.3%, respectively. And GNB3C825T allele T in HUTT-positive and negative groups was 38.3% and 18.7%, respectively (χ(2) = 4.905, P < 0.05). GNB3C825T allele frequency did not show statistically significant difference among the 3 hemodynamic patterns of VVS (χ(2) = 0.658, P > 0.05).</p><p><b>CONCLUSION</b>Study on GNB3C825T allele frequency in children with vasovagal syncope is of significant value for a better understanding of the pathophysiology of VVS and provide a molecular biologic basis for its mechanisms.</p>

Changes of Activities of Na~+K~+-ATPase,Ca~(2+)Mg~(2+)-ATPase in Erythrocyte Membrane and Blood Viscosity in Children with Essential Hpertension / 实用儿科临床杂志

Objective To study the changes of Na+K+-ATPase,Ca2+Mg2+-ATPase activities in erythrocyte membrane and blood viscosity in children with essential hypertension.Methods The activities of Na+K+-ATPase,Ca2+Mg2+-ATPase in erythrocyte membrane were determined by a colorimetric method.Blood viscosity was measured and analyzed with the statistic analysis SPSS 12.0 software in 50 children from Nov.2004 to Dec.2004 in the people's hospital of guizhou province and adolescents with essential hypertension.Thirty healthy children were collected as control group.Results The activities of Na+ K+-ATPase[(6.12?1.30)?molpi/(gHb?h)and(4.59?1.40)?molpi/(gHb?h)],Ca2+Mg2+-ATPase[(7.46?1.30)?molpi/(gHb?h)and(5.81?1.20)?molpi/(gHb?h)] were lower significantly in hypertension group than those in control group(Pa

Clinical Value of Perioperative Plasma D-Dimer in Pediatric Radiofrequency Ablation / 实用儿科临床杂志

Objective To explore the effects of radiofrequency catheter ablation(RFCA)on the blood coaguable states and the clinical value of perioperative plasma D-dimer.Methods The plasma level of D-dimer was assayed by enzyme-linked immunosorbent assay(ELISA)in blood samples of 30 children who were undertaken RFCA.Blood samples were consecutively obtained before cannulating,after electrophysiologic(EP)study,immediately after RFCA,the second day and the seventh day after RFCA.The centrifuged spead was 3 000 r/min,keep it for 10 minutes to obtain the upper plasma,and the crvopreserve.Results The plasma levels of D-dimer was highest at the time point when RFCA was successfully accomplished and restored to preoperative level in the seventh day after RFCA.There were statistically significant difference in the paried values at different time points(Pa