To establish an EDTA complexation extraction pretreatment combining with GFAAS method for the determination of residual aluminium ion in Huoxiang zhengqi pellets without digestive treatment, systematical investigation was made on sample preparation, and EDTA was used for the complexation extraction of residual aluminium ion in samples. The pH, concentration and volume of extraction solution, the temperature and time of microwave extraction, and graphite furnace temperature program were investigated. The results were compared with the microwave digestion. It was showed that, 0.1 g of sample weight was added in 20 mL 0.05 mol x L(-1) EDTA solution (pH 3.5), followed by heating at 150 degrees C for 10 min in the microwave extraction device. The determination of GFAAS was performed at optimized detection wavelength (257.4 nm) as well as graphite furnace temperature program, the detection limits and quantification limits were 2.37 μg x L(-1) and 7.89 μg x L(-1), respectively. The precision (RSD) was less than 2.3%. The average recovery was 96.9% -101%. The present method is easy, rapid and accurate for the determination of residual aluminium ion in Huoxiang zhengqi pellets.
Aluminum ; chemistry ; isolation & purification ; Drug Contamination ; Drugs, Chinese Herbal ; chemistry ; Edetic Acid ; chemistry ; Graphite ; chemistry ; Spectrophotometry, Atomic ; methods ; Temperature

Objective To explore the potential mechanisms of regulating adiponectin secretion and expression in vivo in rats.Methods To observe the regulation of adiponectin by fasing-refeeding and β-adrenergic agonists, male Wistar rats were fasted for 18 h and allowed to refeed or a β3-adrenergic receptor agonist was infused into refeeding rats.The effects of insulin clamp on adiponectin secretion and expression, including euglycemichyperinsulinemic clamp and hyperglycemic-hyperinsulinemic clamp, were also investigated.Plasma adiponectin level was determined by radioimmunoassay.Adiponectin mRNA expression in adipose tissue of rats was detected by realtime PCR.Results (1) Refeeding 18 h fasted rats increased plasma adiponectin concentration (about 2-fold) and adipose tissue adiponectin expression (about 3-fold), which were completely blocked by administration of β-adrenergic agonist.(2) Hyperinsulinemic clamp increased plasma adiponectin concentration and adiponectin gene expression in adipose tissue.Conclusions Adiponectin secretion and expression are acutely regulated in vivo by nutritional status.Insulin and β-adrenergic agonists regulate adiponectin secretion and expression in adipose tissue.

Objective To establish the reference value of liver virtual touch tissues quantification (VTQ) values in healthy subjects. Methods Liver stiffness was measured with Siemens Acuson S2000 ultrasound system in totally 300 healthy subjects. The first one hundred healthy subjects received VTQ measurements in four parts (superficial and deep parts of right lobe, superficial and deep parts of left lobe). Then the achievement rate in different part of liver was calculated to choose the suitable measuring position. On the other hand, the reproducibility was analyzed with intraclass correlation coefficient (ICC). The last two hundred healthy subjects received VTQ measurements in the suitable position only. The reference value of VTQ was calculated using (x-)±1.96s. Results It was stable to measure liver stiffness with VTQ technique. The achievement rate was high in right lobe, and the deep parts of right lobe was the best measuring position. There was significant difference of VTQ value between males and females (P＜0.001), while the VTQ value was similar in different age groups. The reference value was 0.79-1.57 m/s in males and 0.74-1.40 m/s in females. Conclusion Liver VTQ value in healthy subjects are different between males and females.

Objective To investigate the influence of atorvastatin in methylation and expression level of Bcl-2 in human umbilical endothelial cells(HUVECs)treated with homocysteine(Hcy)and to expound potential mechanism of atorvastatin resisting arteriosclerosis.Methods After HUVECs were treated with 0, 2, 4, 8, 16, and 32 mmol·L-1 Hcy for 48 h,MTT was used to measure the inhibitory rates of HUVECs and the half inhibitory concentration (IC50 ). According to the experimental results, the HUVECs cultured in vitro were divided into control group (0.00 mmol · L-1 Hcy ), Hcy group (9.00 mmol·L-1 Hcy ), and atorvastatin group (9.00 mmol·L-1 Hcy+1×10-3 mmol·L-1 atorvastatin).After treated for 48 h,flow cytometry was used to detect the apoptotic rate of cells, the mRNA expression of Bcl-2 was analyzed by fluorescence quantitative PCR,the protein expression of Bcl-2 was detected by Western blotting method, and the methylation level of Bcl-2 promoter region was determined by nest touch-down PCR combined with methylation specific PCR (MSP ). Results Compared with control group,the apoptotic rate of HUVECs in Hcy group was increased(P<0.01),the mRNA and protein expression levels of Bcl-2 were significantly decreased(P<0.01),and the Bcl-2 promoter region methylation level was also decreased(P<0.01).Compared with Hcy group,the apoptotic rate of HUVECs in atorvastatin group was decreased(P<0.01),the mRNA and protein expression levels of Bcl-2 gene were increased (P<0.05), and the Bcl-2 promoter region methylation level was also increased (P<0.05). Conclusion Atorvastatin can prevent the apoptosis of HUVECs induced by Hcy through regulating Bcl-2 methylation.

Objective To construct a recombinant Bacillus Calmette-Guerin ( BCG ) vaccine strain, rBCG::Rv3478-pMV261, expressing the Rv3478 protein of Mycobacterium tuberculosis and to inves-tigate its immunogenicity.Methods The gene fragments encoding Rv3478 antigen were amplified by PCR and then respectively cloned into pMV261 and pET-28a vectors to construct the recombinant expression plas-mids (Rv3478-pMV261 and Rv3478-pET-28a).The Rv3478-pMV261 plasmids were transformed into the BCG cells to construct the rBCG vaccine strains, while the Rv3478-pET-28a plasmids were transformed into Escherichia coli BL21 strains for the expression of Rv3478 protein.Polyclonal antibodies were induced in mice upon the immunization with Rv3478 protein.The rBCG vaccine strains overexpressing Rv3478 protein were screened out with Western blot assay.The C57BL/6 mice were divided into four groups including the PBS treated group, BCG treated group, rBCG::pMV261 ( R0) treated group and rBCG::Rv3478-pMV261 ( R3) treated group.All mice were sacrificed in 4 or 12 weeks after immunization.Enzyme-linked immunos-pot assay ( ELISPOT) , ELISA and flow cytometry analysis were performed to evaluate the induced humoral and cell-mediated immune responses in mice.Results The Rv3478 protein was successfully expressed and could induce polyclonal antibodies in mice.High levels of IFN-γand TNF-αwere detected in mice treated with R3, indicating that the immunization with R3 enhanced the cellular immunity.Moreover, the ratios of CD4+to CD8+T cells and the percentages of CD44+CD62L+T cells were increased in mice upon the immuni-zation with R3.Conclusion The recombinant BCG vaccine strain overexpressing Rv3478 protein could in-duce stronger cell-mediated immune responses in mice.It might be have a great significance as a new tuber-culosis( TB) vaccine strain against TB infection in the future.

Objective To evaluate protective effects of recombinant human thioredoxin(TRX) in myocardial injury of mice with viral myocarditis. Methods We established viral myocarditis models by intraperitoneal injection with 0.1 ml 100TCID50 Coxsackie virus 3m(CVB3m), along with tail vein injection of recombinant human TRX (2 mg/kg) for protection. The control group was given equivalent volume of normal saline. The mice were killed 7 days following the injections. Serum lactate dehydrogenase (LDH) activity was determined and myocardial injury was examined with light microscopy. Results The somm LDH activity in Coxsackie virns-infected mice [(3130.50±390.57)U/L] was higher than that of animals in the control group[ (1617.86±155.42)U/L] and that of TRX protection group[ (1959.43±540.75)U/L], the difference being statistically significant (P<0.05); there was no significant difference between TRX protection group and the control group(P 0.05). Light microscopy showed that five of the eight Coxsackie rims-infected mice had myocardial lesions, including focal myocardial necrosis and inflammatory infiltration. There was no myocardial injury in the TRX protection group. Conclusions Recombinant human TRX can lessen myocardial injuries induced by infection with CVB3m, and so can protect myocardium.

This study aims to investigate the serotype distribution of non-polio enterovirus (NPEV) isolated from patients with acute flaccid paralysis (AFP) during 2011-2012 in Hebei Province, China and to analyze the relationship between these viruses and AFP. NPEV strains were isolated from the stool specimens from AFP cases in Hebei using human rhabdomyosarcoma cells (RD) and the mouse cell line expressing the gene for the human cellular receptor for poliovirus (L20B) according to the WHO requirements. The nucleotide sequence of VP1 region was determined, and the serotypes of NPEV were identified by molecular typing. The results showed that among the 82 strains of NPEV isolated from the AFP cases during 2011-2012, 42 isolates (55.3%) were identified as human enterovirus A (HEV-A), which were classified into 4 serotypes, 34 (44.7%) as human enterovirus B (HEV-B), which were classified into 13 serotypes, 2 as adenovirus, and 4 were untyped; human enteroviruses C and D were not found in these cases. Enterovirus A71 (EV-A71) was the main type of HEV-A, accounting for 85.7% of all HEV-A strains. HEV-A, especially EV-A71, was predominant among the NPEV strains isolated from AFP patients during 2011-2012 in Hebei Province.
Acute Disease ; China ; epidemiology ; Enterovirus ; classification ; physiology ; Humans ; Paralysis ; epidemiology ; virology ; Seasons ; Serotyping

OBJECTIVE: To explore the effect of nitric oxide (NO) on the gene expression of hepatic TNF-α and IL-1β by crush injury of rat's soft tissues. METHODS: Rats were randomly divided into sham group, crush group, crush+aminoguanidine (AG) group, and crush+L-arginine (L-Arg) group. Activities of ALT and AST as well as NO level in serum were measured. Gene expressions of TNF-α and IL-1β were detected with RT-PCR. RESULTS: Obvious increase in TNF-α and IL-1β mRNA expression was detected in the crush group compared with the sham group (P<0.05). After pretreated L-Arg, expressions of TNF-α and IL-1β mRNA were markedly increased (P<0.05). After pretreated AG, those indices obviously decreased (P<0.05). Activities of ALT and AST enhanced and NO level increased in the crush group compared with the sham group (P<0.05). Pretreatment with L-Arg or AG led to substantial increased or reduced activities of ALT and AST as well as NO levels, respectively. CONCLUSION Endogenous NO mediated TNF-α, IL-1β mRNA up expression in liver induced by increased production of NO after crush injury of rat's soft tissues.
Animals ; Gene Expression ; Interleukin-1beta/metabolism* ; Liver ; Nitric Oxide/physiology* ; RNA, Messenger ; Rats ; Tumor Necrosis Factor-alpha/metabolism* ; Wounds and Injuries

Objective To evaluate the utility of MMP9,MPO and sCD40L in detection of the character of coronary artery plaque.Methods From April 2008 to January 2010,118 patients from outpatient of Fu Wai Hospital with chest pain were enrolled.All of them underwent 64 Multiple-detector row spiral computer tomography (64-MDCT),the CT value ＜ 130 Hu patients were enrolled in non-calcified plaque group (71 cases),CT value ≥ 130 Hu patients were enrolled in the calcified plaque group (47 cases).Ninty healthy volunteers were selected as the control group.Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum markers,including MMP9,MPO and sCD40L.Levels of MMP9,MPO and sCD40L of each group were compared.ROC curve was used to evaluate the sensitivity and specificity of the markers in diagnosis of non-calcified plaque.Results MMP9,MPO and sCD40L levels of non-calcified were ( 762.25 ± 368.71 ),[ 844.10 (582.00 - 1220.70) ],(9.37 ± 3.15) μg/L,higher than the healthy control group (342.70 ± 178.53),[426.35 ( 283.20 - 592.00) ],(6.55 ± 2.96) μg/L and calcified plaque group ( 483.12 ± 219.09 ),[ 469.00 ( 302.45 - 723.55) ],( 7.24 ± 2.86) μg/L The difference was statistically significant ( F =42.47,H =50.28,F =17.94,all P ＜ 0.01 ). Areas of MMP9,MPO and sCD40L under the ROC curve to predict non-calcified plaque were 0.854,0.792,0.751 respectively,when the identification threshold for non-calcified plaque were 510.13,537.82,7.05 μg/L respectively,the diagnostic sensitivity was 80％,80％,80％ respectively,and specificity was 80％,67％ and 55％ respectively.Conclusion The serum MMP9,MPO and sCD40L levels can help to determine the　character of coronary plaque.

The complete chloroplast genome of medicinal plant Asarum caudigerum Hance and its close relative A. cardiophyllum Franchet were sequenced using Illumina Hiseq technology, and assembled, annotated, and characterized by bioinformatic methods in this study. Then phylogenetic analysis of the complete chloroplast genomes of A. caudigerum, A. cardiophyllum, and twelve published species was conducted. The results indicated that the chloroplast genomes ranged from 186 215-186 985 bp in length, with a large single copy (LSC, 89 445-90 169 bp) and two inverted repeats (IRa/IRb, 48 387-48 408 bp). The overall GC content was 37.4%-37.5%. A total of 144 chloroplast genes were annotated, including 98 protein coding genes, 38 tRNA genes and 8 rRNA genes. In addition, complex genomic rearrangements were detected in the chloroplast genome of Asarum. Meanwhile, visual evaluation of the discrete type of the sequence indicated that the variation level of non-coding region was higher than that of coding region. Phylogenetic analyses suggested that A. caudigerum and A. cardiophyllum were clustered into a single clade and A. cardiophyllum, A. sieboldii var. seoulense, A. misandrum and A. maculatum were clustered into another single branch. These two clade were sister species. This study provides a scientific basis for the identification, phylogenetic relationship, molecular breeding of Asarum species.