Objective To develop a method for the determination of berberine hydrochloride,palmatine hydrochloride and jatrorrhizine hydrochloride in Yiqi Zhixue Granules. Methods The samples were extracted by ultrasonication with methanol for 30 min,and an ion pair RP-HPLC method was applied to determine three kinds of alkaloids.The column of C18 with temperature at 30 ℃ was used to separate the target components,the mobile phase consisted of the mixed solution of 0.01 mol/L sodium-heptanesulfonate solution and equal volume of 0.01 mol/L potassium dihydrogen phosphate solution(pH being adjusted at 3.1 with phosphoric acid) combined with acetonitrile(70 ∶ 30),the flow rate was 1.0 mL/min,and the detection wavelength was at 345 nm. Results Three kinds of alkaloids were separated perfectly,the average recoveries(n=6)were 100.95 %(RSD=2.10 % ) for berberine chloride,102.14 %(RSD=2.29 % ) for palmatine chloride,and 100.71 %(RSD=2.65 % ) for jatrorrhizine chloride. Conclusion The developed method is demonstrated to be simple,specific and accurate,which can be used to determine the contents of berberine chloride,palmatine chloride and jatrorrhizine chloride in Yiqi Zhixue Granula,and to control the quality of Folium Mahoniae in Yiqi Zhixue Granules.

The safflower floret is a traditional Chinese medicine used to promote blood circulation and remove obstruction in the channels. The spines on its bracts are considered a handicap when manual harvest is involved. In this study, cDNA-SRAP was used to systematically investigate which genes are associated with the spines. Sixty pairs of possible primer combinations were used on two cDNA pools representing spininess and spinelessness. Six transcript-derived fragments were identified, of which two with low recombination were sequenced successfully and named as GPY-1 and GPY-2. By using the RACE method, the full-length cDNA of GPY-2 is cloned and named as CTL-spn. The full-length cDNA of CTL-spn was 1 679 bp long with a 1 524 bp ORF encoding a 508 aminoacid protein. The deduced amino acid sequence of the CTL-spn gene shared a high homology (97%) with other known ATP synthase CF1 alpha subunits. Semiquantitative RT-PCR analysis revealed that the mRNA of GPY-1 and GPY-2 accumulated in only spiny lines. Considering the important role of ATP synthase CF1 alpha subunit in plants, it may directly take part in the formation process of spininess and enhancing resistance reaction of spiny safflower. Also, our results provide the important insights for breeding spineless cultivars of safflower.
Adenosine Triphosphate ; Amino Acid Sequence ; Carthamus tinctorius ; enzymology ; genetics ; Chloroplast Proton-Translocating ATPases ; genetics ; DNA Primers ; DNA, Complementary ; Plant Proteins ; genetics

OBJECTIVETo explore the effects of trichloroethylene (TCE) on cardiac developmental differentiation in human embryonic stem cells.

METHODSIn this study, based on the human embryonic stem cells in vitro cardiac differentiation assay, we investigated the potential effect of TCE exposure on the cardiac toxicity in embryo development. Human embryonic stem cells were treated with TCE at different concentrations of 100 ppb, 1 ppm, and 10 ppm and dimethyl sulfoxide(DMSO) treated as control. The MTT assay was performed to examine the cytoplasmic toxicity of TCE exposure. The beating percentages were recorded and the expression of cardiac specific gene was evaluated by PCR or flow cytometry. Also, real time PCR was performed to verify the micro array analysis on the expression level changes of genes which were involved in the Ca2+ signal pathways.

RESULTSCompared with the control group, there was no significant difference in cell viability when cells were treated with TCE at the concentrations of 100 ppb, 1 ppm, and 10 ppm. However, TCE could inhibit the expression of cTnT protein in a concentration-dependant manner. And the most interestingly, TCE significantly inhibited the cardiac differentiation characterized by the decrease beating percentages. Genes involved in Ca2+ signaling pathway were severely disrupted by TCE.

CONCLUSIONTCE inhibited the cardiac specific differentiation of human embryonic stem cell and at the meanwhile the genes responsible for Ca2+ signaling pathway were severely disrupted, which could contribute the severe effects of TCE cardiotoxicity.

Calcium Signaling ; Cell Differentiation ; Cells, Cultured ; Embryonic Development ; Embryonic Stem Cells ; cytology ; drug effects ; Heart ; embryology ; Humans ; Trichloroethylene ; toxicity

Hot Temperature ; Humans ; Occupational Exposure ; Occupational Medicine ; standards

This study aimed to amplify major genome segments (VP7, VP4, VP6, VP2 and NSP2-5) of porcine G3 group A rotavirus (GARV) LLZ212 isolated in our laboratory, determine their genotypes, and explore the evolutionary relationships between G3 GARV strains isolated from humans and pigs in Lulong County, Hebei Province, China. Major genome segments of seven GARV strains were amplified by reverse transcription-polymerase chain reaction (RT-PCR) and the segments were sequenced. The genome segments of seven GARV strains were determined by the online RotaC genotyping tool (RotaC v2.0). The reference sequences of each GARV genome segment were downloaded from GenBank. Homology and phylogenetic evolutionary analyses were conducted using the MEGA 5.0 and DNAStar software packages. LLZ212 isolated from pigs in Lulong had the following genotype: G3-P[8]-I5-C1-N1-T1-E1-H1. All human GARV strains had the following genotype: G3-P[8]-I1-C1-N1-T1-E1-H1. The VP7, VP4, NSP4 and NSP5 genes of the LLZ212 strain had the highest nucleotide identities with the human GARV E885, CMH014/07, Wa and RMC321 strains, respectively, and these clustered together in a sublineage. The VP6, NSP4 and NSP5 genes of the LLZ212 strain shared the highest nucleotide identities with the porcine GARV PRG921 strain, while VP2 associated most closely with porcine GARV OSU strain, and these also clustered in a sublineage. A rare porcine G3-P[8]-I5-C1-N1-T1-E1-H1 GARV strain was identified, which may represent a reassortment between porcine and human viruses. In conclusion, the VP7, VP4, NSP4 and NSP5 genes of LLZ212 share high levels of sequence identity with human GARV, while VP2, VP6, NSP2 and NSP3 cluster with porcine GARV.
Animals ; Capsid Proteins ; genetics ; Child, Preschool ; China ; epidemiology ; Evolution, Molecular ; Genotype ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny ; Rotavirus ; classification ; genetics ; isolation & purification ; Rotavirus Infections ; epidemiology ; veterinary ; virology ; Swine ; Swine Diseases ; epidemiology ; virology ; Viral Nonstructural Proteins ; genetics

The study aims to develop a unified method to determine seven phenolic acids (neochlorogenic acid, chlorogenic acid, 4-caffeoylquinic acid, caffeic acid, isochlorogenic acid B, isochlorogenic acid A and isochlorogenic acid C) contained in honeysuckle flower that is the monarch drug of all the eight Yinqiao Jiedu serial preparations using quantitative analysis of multi-components by single-marker (QAMS). Firstly, chlorogenic acid was used as a reference to get the average relative correction factors (RCFs) of the other phenolic acids in ratios to the reference; columns and instruments from different companies were used to validate the durability of the achieved RCFs in different levels of standard solutions; and honeysuckle flower extract was used as the reference substance to fix the positions of chromatographic peaks. Secondly, the contents of seven phenolic acids in eight different Yinqiao Jiedu serial preparations samples were calculated based on the RCFs durability. Finally, the quantitative results were compared between QAMS and the external standard (ES) method. The results have showed that the durability of the achieved RCFs is good (RSD during 0.80% - 2.56%), and there are no differences between the quantitative results of QAMS and ES (the relative average deviation < 0.93%). So it can be successfully used to the quantitative control of honeysuckle flower principally prescribed in Yinqiao Jiedu serial preparations.
Caffeic Acids ; analysis ; Chlorogenic Acid ; analogs & derivatives ; analysis ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Flowers ; chemistry ; Hydroxybenzoates ; analysis ; Lonicera ; chemistry ; Quinic Acid ; analogs & derivatives ; analysis

PTP4A3 is an oncogene,which encodes phosphatase of regenerating liver(PRL)-3 protein that is a metastasis-associated phosphorylase. At present,a number of studies have found that it promotes cell proliferation, cell motility,cell invasion,tumor metastasis and epithelial-mesenchymal transition. Therefore,it is inseparable with the occur-rence and development of cancer. Here,we summarize the relationship between PTP4A3 gene and the occurrence and de-velopment of cancer,the alteration of PTP4A3 gene expression and the functional role of PTP4A3 gene in a variety of canc-ers. This review will help us to understand the correlation between PTP4A3 gene and cancer as well as the mechanism of signaling pathway,providing new insights of PTP4A3 gene targeting strategy for treating cancer.

This study aimed to study the epidemiological and clinical characteristics of human bocavirus 1-4 (HBoV1-4) in children with acute diarrhea in Lanzhou and to investigate the association between HBoV and acute gastroenteritis. A total of 331 stool samples were collected from children aged under 5 years with acute diarrhea at the Department of Pediatrics, the First Hospital, Lanzhou University, between July 2012 and June 2013. Nested PCR was used to screen for HBoV and a general PCR was employed to screen other common diarrhea viruses. We found human bocavirus 1, 2, 3 and 4 in 26, 15, 7 and 1 cases, respectively. There was no specific seasonal distribution of HBoV, with infections occurring throughout the year. HBoV was mostly found in children aged between 7 and 12 months, with a mean age of 11.04 months (+/- 6.92 months), and 93.88% of affected children were aged under 2 years. Overall, 71.3% of mixed infections were mixed and the majority of other infections were caused by rotavirus. There was no statistical difference in the incidence of fever and vomiting associated with HBoV infection. A rare virus strain, HBoV4 (LZFB086), was identified, which showed highest levels of nucleotide sequence identity (99.0%) with a single Thai HBoV strain (JQ267789). No case of HBoV2B was found. In conclusion, HBoV1 was a major etiological pathogen of HBoV in pediatric cases in Lanzhou. HBoV4 was detected in feces for the first time in China. The rate of mixed infections was high and rotavirus was dominant. The data presented suggests that HBoV is not a major causative agent of gastroenteritis.
China ; epidemiology ; Diarrhea ; epidemiology ; virology ; Feces ; virology ; Human bocavirus ; classification ; genetics ; isolation & purification ; Humans ; Infant ; Molecular Sequence Data ; Parvoviridae Infections ; epidemiology ; virology ; Phylogeny ; Seasons

Adult ; Aged ; Aged, 80 and over ; Alcoholism ; Diabetes Mellitus ; epidemiology ; etiology ; Female ; Hepatic Encephalopathy ; epidemiology ; etiology ; Hepatitis B ; complications ; pathology ; Hepatitis B virus ; Humans ; Liver Cirrhosis ; etiology ; pathology ; Liver Cirrhosis, Alcoholic ; etiology ; pathology ; Liver Function Tests ; Liver Neoplasms ; epidemiology ; etiology ; Male ; Middle Aged ; Prognosis ; Retrospective Studies

Abstract:This study aims to investigate the genetic characteristics of group A rotavirus (GARV) G9P[8] strains from infantile diarrhea samples in Hebei Lulong region from 2009 to 2011. We randomly selected five GARV G9P[8] strains in Hebei Lulong region from 2009 to 2011, amplified the 11 gene fragments of GARVs by RT-PCR, and analyz their full-genome sequences by homology and phylogenetic analysis with DNAStar and MEGA. The nucleotide homology between strains LL11131077 and LL11131083 in 2011 was significantly higher than hat etween them and the other three strains in 2009 and 2010. The G9P[8] GARVs circulating in Hebei Lulong region from 2009 to 2011 elenged to the same genotype as the prevalent G9P[8] GARVs in other parts of the world. However,the two strains in 2011, compared with those in 2009 and 2010, were located in a different sub-branch of the phylogenetic tree and had amino acid mutations at many sites.
China ; Feces ; virology ; Genome, Viral ; Genotype ; Humans ; Molecular Sequence Data ; Phylogeny ; Rotavirus ; classification ; genetics ; isolation & purification ; Rotavirus Infections ; virology ; Viral Proteins ; genetics