Study on HPLC Fingerprint of Qingbi Granules / 中国药房

OBJECTIVE:To establish HPLC fingerprint of Qingbi granules.METHODS:HPLC method was adopted.The determination was performed on Diamonsil C18 column with mobile phase consisted of methanol-0.3％ phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 260 nm,and column temperature was 25 ℃.The sample size was 10 μL.Using baicalin as reference,HPLC chromatograms of 10 batches of samples were determined.Common peak identification and similarity evaluation were performed by using TCM Chromatographic Fingerprint Similarity Evaluation Software (2.0 edition).RESULTS:There were 29 common peaks in HPLC chromatograms of 10 batches of samples.The similarity among the 10 batches was more than 0.90.After validation,HPLC chromatograms of 10 batches of samples were in line with control fingerprints.CONCLUSIONS:Established fingerprints can provide reference for identification and quality evaluation of Qingbi granules.

Content Determination of Verbascoside in Naosaitong Honeyed Pill by HPLC / 中国药房

OBJECTIVE:To establish a method for the content determination of verbascoside in Naosaitong honeyed pill. METHODS:HPLC was performed on the column of Zorbax ODX with mobile phase of acetonitrile-0.1% phosphoric acid(16∶84, V/V)at a flow rate of 1.0 ml/min,the detection wavelength was 334 nm,column temperature was 30 ℃,and the injection volume was 10 μl. RESULTS:The linear range of verbascoside was 0.200-1.000 μg(r=0.999 8);RSDs of precision,stability and repro-ducibility tests were lower than 1%;recovery was 95.16%-99.78%(RSD=1.80,n=6). CONCLUSIONS:The method is simple and accurate,and suitable for the content determination of verbascoside in Naosaitong honeyed pill.

Biomass fast pyrolysis for bio-oil production in a fluidized bed reactor under hot flue atmosphere / 生物工程学报

Fast pyrolysis experiments of corn stalk were performed to investigate the optimal pyrolysis conditions of temperature and bed material for maximum bio-oil production under flue gas atmosphere. Under the optimized pyrolysis conditions, furfural residue, xylose residue and kelp seaweed were pyrolyzed to examine their yield distributions of products, and the physical characteristics of bio-oil were studied. The best flow rate of the flue gas at selected temperature is obtained, and the pyrolysis temperature at 500 degrees C and dolomite as bed material could give a maximum bio-oil yield. The highest bio-oil yield of 43.3% (W/W) was achieved from corn stalk under the optimal conditions. Two main fractions were recovered from the stratified bio-oils: light oils and heavy oils. The physical properties of heavy oils from all feedstocks varied little. The calorific values of heavy oils were much higher than that of light oils. The pyrolysis gas could be used as a gaseous fuel due to a relatively high calorific value of 6.5-8.5 MJ/m3.

Effect of rapamycin on high glucose-induced autophagy impairment, oxidative stress and premature senescence in rat mesangial cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of rapamycin and 3-methyladenine on autophagy impairment, oxidative stress and premature senescence induced by high-glucose in primarily cultured rat mesangial cells.</p><p><b>METHODS</b>Rat glomerular mesangial cells (GMCs) were isolated and cultured in normal glucose, high glucose, high glucose with 3-methyladenine (3-MA), or high glucose with rapamycin. At 24 h, 72 h and 10 days of culture, the cells were examined for expression levels of autophagy markers LC3 and p62/SQSTM1, malondialdehyde (MDA) and protein carbonyl, β-galactosidase (SA-β-gal) activity and heterochromatin foci (SAHF).</p><p><b>RESULTS</b>Compared with those of normal cell culture, the cells exposed to high glucose for 72 h and 10 days showed down-regulated LC3 expression, up-regulated p62/SQSTM1 expression, elevated MDA and protein carbonyl levels, and increased SAHF formation and percentage of SA-β-gal-positive cells. These changes were reversed in GMCs exposed to high glucose and rapamycin for 72 h and 10 days, but exacerbated in cells incubated with 3-MA.</p><p><b>CONCLUSION</b>High glucose can suppress autophagic function of rat GMCs to result in oxidative damage and cell senescence. Rapamycin can attenuate autophagy impairment, oxidative damage and senescence induced by high glucose, whereas 3-MA can further aggravate high glucose-induced cell injuries in rat GMCs.</p>

Effects of NaDC3 over-expression on energy metabolism and ROS generation in human renal tubular cells / 中国病理生理杂志

AIM: To study the effects of over-expression of tricarboxylic acid cycle intermediates transporter NaDC3 (high affinity sodium-dependent dicarboxylate transporter) on energy metabolism and ROS generation in human renal tubular cells. METHODS: Recombinant retrovirus vector containing NaDC3 gene was constructed and used for infecting human renal tubular epithelial cell HKC. Control vector containing Neo gene was also constructed and infected cells. Liquid scintillation method was used to determine the level of [~3H]-succinate (as a transport substrate of NaDC3) in the cells. Clark electrode method and reverse phase HPLC were used to detect oxygen consumption and ATP content intracellularly, respectively. Mitochondrial membrane potential and reactive oxygen species (ROS) content in HKC were determined with laser confocal microscope after treatment with fluorescent probe JC-1 and CM-H_2DCFDA, respectively. RESULTS: Western blotting analysis showed that the expression of NaDC3 protein in uninfected- and control vector-infected cells was at lower level. After infection with recombinant NaDC3 vector, expression level of NaDC3 protein in HKC cells was increased markedly. Transport assay revealed that the level of [~3H]-succinate in NaDC3-infected cells was noticeably increased. Oxygen consumption and ATP content in NaDC3-infected HKC were significantly higher than those in uninfected- and control vector-infected cells. Laser confocal analysis revealed that mitochondrial membrane potential and ROS level in NaDC3-infected HKC were increased, compared with uninfected- and control vector-infected cells. CONCLUSION: Over-expression of NaDC3 accelerates the speed rate of energy metabolism and increases intracellular ROS generation by transporting an overdose of tricarboxylic acid cycle intermediates in human renal tubular epithelial cells.

TRIMETHYLGLYCINE COULD OVERCOME THE DIFFICULTY IN PCR AMPLIFICATION OF G+C-RICH GENES / 解放军医学杂志

To explore the effects of different organic solvents including trimethylglycine on the PCR amplification of G+C rich DNA templates,two high G+C containing DNA templates (cyclin dependent kinase inhibitor gene ARF and IGFR II) were amplified using standard PCR and modified PCR with the addition of glycerin, dimethylsulfoxide (DMSO), formamide and trimethylglycine respectively. There were no specific amplified products, when standard PCR and modified PCR with the addition of glycerin, DMSO and formamide were used to amplify the G+C rich ARF and IGFR II genes. But the two genes could be specifically amplified with the addition of optimal concentration of trimethylglycine in the PCR mixture. These results indicate that trimethylglycine can abolish or reduce the formation of second structures in G+C rich DNA regions and promote the amplification of high G+C containing DNA templates.

Effects of oxidative stress and its secondary protein impairment induced by high glucose content in human renal tubular epithelial cell / 解放军医学杂志

Objective To explore the effects of oxidative stress and its secondary protein impairment induced by high glucose content in human kidney proximal tubular epithelial cell line (HKC). Methods Cultured HKC were used to detect the reactive oxygen species (ROS) and protein carbonyl content. HKC were divided into three different groups: control group (normal glucose), short-term treatment with high glucose group and long-term treatment with high glucose group. The glucose concentrations in control group and the high glucose treatment groups were 5.5mmol/L and 30mmol/L, respectively. Six time-points were set in short-term treatment with high glucose group namely 30min, 1h, 6h, 12h, 24h and 48h; glucose treatment was instituted for 2 months in long-term treatment of high glucose group. 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate, acetyl ester (CM-H2DCFDA) was used as ROS trapping agent, and flow cytometry and laser confocal microscopy were used to detect the concentration of ROS in HKC after treatment with high glucose for different periods of time. Flourescent probe JC-1 was used to test the change in mitochondrion membrane potential (MMP). The protein carbonyl content was determined by 2,4-dinitrobenzene hydralazine (DNPH) chromatometry. Results The ROS level in HKC reached the peak at 1 hour after treatment with high glucose, and it then decreased gradually (P0.05), but after 2 months, there was significant change in the content of DNPH (P

The effects of human Na~+-dependent high-affinity dicarboxylate cotransporter 3 and glucose on the transport of succinate in human proximal tubular epithelial cells / 解放军医学杂志

Objective To investigate the effects of Na +-dependent high-affinity dicarboxylate cotransporter 3(NaDC3) and glucose on the transport of succinate in human proximal tubular epithelial cells (HKC) . Methods Western blot was used to detect the expression of hNaDC3 protein in the pcDNA3-hNaDC3, pcDNA3-AhNaDC3 and pcDNA3, transfected HKC, and normal HKC cell. Analysis of succinate contents in the intaking-buffer at the various time points and in the variouse concentrations of glucose were determined by capillary zone electrophoresis. Results The cell transfected with pcDNA3- hNaDC3 was found to have over-expression of hNaDC3 protein for about 1.5 times that of the normal HKC, while succinate content in intaking-buffer was decreased. Although the cells transfected with pcDNA3-AhNaDC3 showed lower expression of hNaDC3 protein, being about 0.6 times that of normal HKC cells, the decrease of succinate in intaking buffer was not obviously effected. There were no differences in hNaDC3 protein content and succinate intaking between the cells transfected with pcDNA3 and normal HKC cells. With the increase in glucose concentration, the decrease of succinate in intaking-buffer was accelerated. Conlusion The results suggested that the overexpression of hNaDC3 protein can cause the succinate-intake more quickly in HKC.

UASLG identification of Rana temporatia chensinensis David's / 中成药

AIM: To apply UASLG for Rana temporatia chensinensis David's identification. METHODS: Rana temporatia chensinensis David was made ultrasonic extraction. These solvents in turn were petroleum ether、chlorofo mn、ethyl acetate、n-butanol and 95%WTBZ# ethanol,then their absorbance in UV and one-order derivative spectrums were determined. RESULTS: Rana temporatia chensinensis David's UASLG was reproducible. CONCLUSION: UASLG's action spectrums for Rana temporatia chensinensis David can be identified for quality control.

Effect of p19ARF on growth and apoptosis of leukemia cells / 中国病理生理杂志

AIM: To study the effect of p19ARF on the biological behavior of human leukemia cells. METHODS: p19ARF was cloned in eukaryotic expression vector pcDNA3.1 and transferred into INK4a/ARF locus-deficient leukemia cells HEL and K562. The changes in biological characteristics of the two p19ARF-transfected cells were observed. RESULTS: The growth of the p19ARF-transfected HEL cells was significantly inhibited compared with the vector-transfected cells; Cell cycle analysis showed that the expression of foreign p19ARF gene resulted in G1 phase cell cycle arrest and apoptosis cell death in some of HEL cells. However, p19ARF had no marked effects on the growth of K562 cells with p53 gene mutation and did not induce apoptosis in K562 cells. CONCLUSION: p19ARF suppressed the growth of leukemia cells by p53 -dependent pathway.

CHANGES OF ? ADRENERGIC RECEPTORS AND ANGIOTENSIN Ⅱ RECEPTORS IN SPONTANEOUSLY HYPERTENSIVE RATS AND DIFFERENT AGES WKY RAT HEARTS / 解放军医学杂志

To study the changes of ? adrenergic receptor subtypes (? 1 AR,? 2 AR) and angiotensin Ⅱreceptor subtypes (AT 1 R,AT 2 R)mRNA levels in spontaneously hypertensive rat (SHR) hearts at 14 weeks of age, and WKY rat hearts at ages 14 and 48 weeks, the total cellular RNA was extracted from the left ventricular tissue, and reverse polymerase chain reaction analysis was performed for measurement of the four receptors. The expression levels of ? 1 AR mRNA and ? 2 AR mRNA were down regulated in WKY hearts at age 48 weeks and in 14 week SHR. AT 1 R mRNA levels were decreased in 48 week WKY and 14 SHR, and AT 2 R mRNA levels were significantly increased in SHR. The expression levels of ? 1 AR and AT 1 R mRNA were declined following the increase of ages. The levels of ? 2 AR mRNA were down regulated in hypertension and myocardial hypertrophy conditions first because ? 2 AR was more sensitive to myocardial hypertrophy than ? 1 AR. Long term and hyper AngⅡ levels stimulated AT 1 R mRNA down regulation in SHR, and it is the myocardial hypertrophic compensatory process that increases the AT 2 R mRNA levels.