1.Correlation of gene expression related to amount of ginseng saponin in 15 tissues and 6 kinds of ginseng saponin biosynthesis.
Kang-yu WANG ; Mei-ping ZHANG ; Chuang LI ; Shi-cui JIANG ; Rui YIN ; Chun-yu SUN ; Yi WANG
China Journal of Chinese Materia Medica 2015;40(16):3168-3173
Fifteen tissues of 4-year-old fruit repining stage Jilin ginseng were chosen as materials, six kinds of monomer saponins (ginsenosides Rg1, Re, Rb1, Rc, Rb2 and Rd) content in 15 tissues was measured by HPLC and vanillin-sulfuric acid method. The relative expression of FPS, SQS, SQE, OSC, β-AS and P450 genes in 15 tissues was analyzed by real-time PCR. The correlations between ginseng saponin content in 15 tissues of Jilin ginseng and biosynthetic pathway -related genes were obtained. The results showed that was a synergistic increase and decrease trend of positive linear correlation among six kinds of monomer saponin content, and there was a significantly (P < 0.01) positive correlation between monomer saponin content and total saponins content. Monomer saponin content and 6 kinds of enzyme gene correlation were different. Biosynthesis of ginseng total saponins and monomer saponin were regulated by six kinds of participation ginsenoside biosynthesis enzyme genes, the expression of these six kinds of genes in different tissues of ginseng showed collaborative increase and decrease trend, and regulated biosynthesis of ginseng ginsenoside by group coordinative manner.
Drugs, Chinese Herbal
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analysis
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Gene Expression Profiling
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Panax
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chemistry
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genetics
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metabolism
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Plant Proteins
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genetics
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metabolism
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Plant Structures
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chemistry
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genetics
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metabolism
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Plants, Medicinal
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chemistry
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genetics
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metabolism
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Saponins
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analysis
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metabolism
2.Epidemiological characteristics of epidemiologic hemorrhagic fever in Qian'an county, Hebei province from 2004 to 2010
Cui-ling, WANG ; Li-zhu, XIAO ; Zi-tian, FAN ; Jian-yuan, TIAN ; Yin-ping, CHEN ; Ai-min, ZHAO
Chinese Journal of Endemiology 2012;31(1):94-96
Objective To analyze the epidemiologic characteristics of epidemic hemorrhagic fever syndrome (EHF) from 2004 to 2010 and provide scientific basis for formulating control measures.Methods Epidemiological data of EHF between 2004 to 2010 were obtained from the“National Disease Surveillance Report on Management Information System”,and the population data were from the“National Disease Surveillance Information Management System for Basic Information Report System”.Descriptive epidemiological methods was used for statistical analysis.ResultsA total of 173 cases were reported in Qian'an from 2004 to 2010.From 2004 to 2010,the cases were 86,67,12,1,0,1,and 7 cases,respectively,and the rate were 13.12/10 million ( 86/640 249 ),10.42/10 million (67/642 688 ),1.86/10 million ( 12/645 124),0.15/10 million (1/647 983 ),0(0/650 720),0.15/10 million( 1/653 839),and 0.11/10 million(7/657 380),respectively.The overall rate was 3.86/10 million(25/648 283) of population.From 2004 to 2008 the incidence reported declined rapidly,then increased slowly after 2009.The cases were found intensively in winter(November - next January) and spring season (february - may).The incidence in the age group of 10 - 45 was higher than that of other age groups,and the total number of cases was 82.08%(142/173).The incidence in males( 114 cases) was higher than that of females(59 cases).Occupational distribution mainly to peasants and students,which accounted for 87.86% (152/173).Conclusions Epidemic in the city declines rapidly follows by a slow recovery,suggesting that in the future surveillance,mice-killing and protection of vulnerable population should be strengthened.
3.Evaluation of the effects of glycosylation on in vivo survival of cold-storage human platelets by using rabbit model.
Rong-Cai TANG ; Cheng-Yin HUANG ; Xiao-Ping PEI ; Bao-An CHEN ; Cui-Ping LI ; Guang-Yao SHI
Journal of Experimental Hematology 2005;13(6):1113-1116
To study the effects of glycosylation on survival of cold-storage human platelets by using rabbit model. (51)Cr-labeling platelets were used to detect the platelet storage survival. The human platelets (2.0 x 10(12)/L) treated with 5 g/L uridine diphosphate galactose (UDP-Gal) were stored in 4 degrees C refrigeratory up to 10 days. The survival of human platelets in rabbits whose reticuloendothelial system was inhibited by the administration of ethyl palmitate was monitored in blood drawn at various times after the platelet transfusion. The results showed that the survival rate of platelets was significantly increased in cold-storage human platelets by UDP-Gal treatment. The survival rates of platelets at 2 hours after transfusion into rabbits in groups of fresh platelets group, UDP-Gal + cold platelets group and cold platelets group were (68.9 +/- 8.5)%, (65.4 +/- 8.0)% and (5.0 +/- 2.6)%, respectively. Compared with cold platelets group, significant differences were seen among all groups (P < 0.01). UDP-Gal + cold platelets group had no significant differences compared with fresh platelets group (P > 0.05). It is concluded that UDG-Gal can provide the protective effect on cold-storage human platelets and prolong the survival time of refrigerated human platelets in rabbit model.
Animals
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Blood Platelets
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cytology
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metabolism
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Blood Preservation
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Cell Survival
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drug effects
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Cryopreservation
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methods
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Glycosylation
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drug effects
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Humans
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Models, Animal
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Platelet Transfusion
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Rabbits
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Uridine Diphosphate Galactose
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pharmacology
4.Cloning, expression and functional identification of a type III polyketide synthase gene from Huperzia serrata.
Jin-cui YE ; Ping ZHANG ; Jie-yin SUN ; Chao-tan GUO ; Guo-shen CHEN ; Ikuro ABE ; Hiroshi NOGUCHI
Acta Pharmaceutica Sinica 2011;46(10):1273-1278
A cDNA encoding novel type III polyketide synthase (PKS) was cloned and sequenced from young leaves of Chinese club moss Huperzia serrata (Thunb.) Trev. by RT-PCR using degenerated primers based on the conserved sequences of known CHSs, and named as H. serrata PKS2. The terminal sequences of cDNA were obtained by the 3'- and 5'-RACE method. The full-length cDNA of H. serrata PKS2 contained a 1212 bp open reading frame encoding a 46.4 kDa protein with 404 amino acids. The deduced amino acid sequence of H. serrata PKS2 showed 50%-66% identities to those of other chalcone synthase super family enzymes of plant origin. The recombinant H. serrata PKS2 was functionally expressed in Escherichia coli with an additional hexahistidine tag at the N-terminus and showed unusually versatile catalytic potency to produce various aromatic tetraketides, including chalcones, benzophenones, phloroglucinols, and acridones. In particular, the enzyme accepted bulky starter substrates N-methylanthraniloyl-CoA, and carried out three condensations with malonyl-CoA to produce 1, 3-dihydroxy-N-methylacridone. Interestingly, H. serrata PKS2 lacks most of the consensus active site sequences with acridone synthase from Ruta graveolens (Rutaceae).
Acyltransferases
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genetics
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isolation & purification
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metabolism
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Amino Acid Sequence
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Cloning, Molecular
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DNA, Complementary
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genetics
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DNA, Plant
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genetics
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Escherichia coli
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genetics
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metabolism
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Gene Expression Regulation, Plant
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Huperzia
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enzymology
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genetics
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Molecular Sequence Data
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Plant Leaves
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enzymology
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genetics
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Plants, Medicinal
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enzymology
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genetics
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Recombinant Proteins
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genetics
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metabolism
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Sequence Alignment
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Substrate Specificity
5.Development of pulsed field gel electrophoresis and application for characterization and identification of Lactobacillus and Streptococcus thermophilus.
Yin-ping DONG ; Sheng-hui CUI ; Hong-xia YU ; Feng-qin LI
Chinese Journal of Preventive Medicine 2011;45(12):1086-1089
OBJECTIVETo develop a pulsed field gel electrophoresis (PFGE) method for molecular typing of Lactobacillus and Streptococcus thermophilus (S. thermophilus) and to apply it in identification and characterization of both bacteria isolated from yoghurt collected from Beijing supermarket.
METHODSThe five most useful restriction enzymes including Apa I, Not I, Sfi I, Xba I and Sma I were chosen to cut DNA of 52 strains of Lactobacillus, S. thermophilus as well as associated standard bacteria strains. The endonucleases and electrophoresis conditions for PFGE analysis were optimized and applied in molecular typing of Lactobacillus and S.thermophilus isolates. Cluster analysis based on the PFGE data was conducted. The identification results of PFGE were compared with those obtained in biochemical and 16s ribosomal RNA PCR identification tests.
RESULTSNot I was suitable for L. bulgaricus, L. fermentum and L. delbrueckii digestion. While Apa I was an appropriate endonuclease for S. thermophilus, L. acidophilus and L. casei digestion. The results of molecular typing indicated that 24 strains of L.bulgaricus and 15 strains of S. thermophilus were grouped into 8 types by PFGE method, respectively. While 7 strains of L.acidophilus were grouped into 3 types and 2 strains of L. delbrueckii were grouped into 2 different PFGE types.
CONCLUSIONThe results of PFGE analysis are in compliance with those of 16s rRNA PCR and biochemical identification. The PFGE method developed in this study is suitable for molecular characterization of both Lactobacillus and S. thermophilus.
Bacterial Typing Techniques ; methods ; Electrophoresis, Gel, Pulsed-Field ; methods ; Lactobacillus ; classification ; isolation & purification ; Streptococcus thermophilus ; classification ; isolation & purification
6.The changes of resting Ca2+i level and expression of CaM mRNA, CaMPK II mRNA in hippocampal neurons of the mice with vascular dementia.
Pei-yuan LU ; Wen-bin LI ; Yu YIN ; Wei-bin WANG ; Cui-ping LIANG
Chinese Journal of Applied Physiology 2004;20(2):146-149
AIMTo investigate the resting [Ca2]i level and expression of calmodulin (CaM), calmodulin-dependent protein kinase II (CaMPKII) mRNA in hippocampal neurons of the mice with vascular dementia (VD) and their roles in the pathogenesis of VD.
METHODSThe mice were subjected for ischemia/reperfusion repeatedly on bilateral common carotid arteries by knots to establish the VD models. Animals with the sham-operation were taken as control group. The changes of behavior were observed through the step-down avoidance test and water maze test on the day 29, 30 after the operations. The hippocampal neurons were obtained immediately after mice were sacrificed and the resting [Ca2+]i was measured using laser scanning confocal microscopy with Fluo-3/AM as fluorescence indicator. RT-PCR technique was used to measure the mRNA expression of CaM, CaMPKII in hippocampal neurons.
RESULTS(1) The abilities of learning and memorizing in model group were inferior to those of sham-operation group( P < 0.05). (2) The resting [Ca2]i level in model group was significantly higher than sham-operation group (P < 0.05), while the expression of CaMmRNA, CaMPKIImRNA in VD group was significantly reduced than sham-operation group (P < 0.01).
CONCLUSIONOur study indicates that excessive resting[Ca2+ ]i level and lower CaM, CaMPKII expression in hippocampal neurons might participate in the pathogenesis of VD.
Animals ; Calcium ; analysis ; Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; genetics ; metabolism ; Calmodulin ; genetics ; metabolism ; Dementia, Vascular ; metabolism ; Hippocampus ; cytology ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Neurons ; metabolism ; RNA, Messenger ; genetics
7.The changes of cAMP and adenylyl cyclase mRNA level in hippocampus of mice with ischemia/reperfusion.
Wei-Bin WANG ; Chun-Feng SONG ; Pei-Yuan LU ; Yu YIN ; Cui-Ping LIANG
Chinese Journal of Applied Physiology 2005;21(3):278-280
AIMTo observe the cAMP and adenylyl cyclase (AC) mRNA level in hippocampus of mice with vascular dementia (ischemia/reperfusion), and explore the molecular pathogenesis of ischemia/reperfusion.
METHODSThe mice were subjected for ischemia/reperfusion three times on bilateral common carotid arteries by knots to establish models of ischemia/reperfusion and the changes of learning and memory were tested on 29 d/30 d after operation. Sham-operation mice were introduced as control group. The cAMP level was evaluated by the radioimmunoassay (RIA), AC mRNA positive neurons of hippocampus CA1 area were dyed through in-situ hybridization (ISH).
RESULTSCompared with sham-operation group, the learning and memory of model group was worse (P < 0.05), the cAMP level in hippocampus was lower (P < 0.05) and the surface density (Sv) of AC mRNA positive neurons reduced (P < 0.05).
CONCLUSIONThe lower cAMP and AC mRNA level in hippocampus might participate in the molecular pathogenesis of ischemia/reperfusion.
Adenylyl Cyclases ; genetics ; metabolism ; Animals ; Cyclic AMP ; genetics ; metabolism ; Hippocampus ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; RNA, Messenger ; genetics ; Reperfusion Injury ; genetics ; metabolism
9.Transcriptional repression of hDaxx enhanced by adenovirus 12 E1B 55-kDa oncoprotein interacting with hDaxx.
Yan-ping WAN ; Yi-mou WU ; Cui-ming ZHU ; Wei-guo YIN ; Heng-ling CAI ; Min-jun YU
Chinese Medical Journal 2004;117(5):753-757
BACKGROUNDDaxx has been identified as a nuclear protein that involves in apoptosis and transcriptional repression. Daxx co-localizes with the promyelocytic leukemia (PML) protein and regulates transcription. Human Daxx (hDaxx) is a protein that functions as a transcriptional regulation through its interaction with some DNA-associated proteins. The aim of this study was to explore the transcriptional regulatory effect of hDaxx interacting with adenovirus (Ad) 12 E1B (Ad12E1B) 55-kDa oncoprotein.
METHODSThe co-localization of hDaxx-Ad12E1B or hDaxx-PML protein in the nucleus was observed under a confocal microscope. Interaction of hDaxx and Ad12E1B was analyzed by yeast two-hybrid assay. Direct binding of hDaxx and Ad12E1B was analyzed using coimmunoprecipitation and Western blot in vivo and in vitro. The activity of a luciferase reporter gene, which was regulated by an hDaxx modulated thymidine kinase (TK) promoter, was detected in an automat luminometer.
RESULTSAd12E1B, which co-localized with hDaxx in the nuclei of G401-CC3 cells, disrupted the co-localization of hDaxx and PML in the PML oncogenic domains (PODs). hDaxx bound directly to Ad12E1B in vivo and in vitro. hDaxx interacted with Ad12E1B along its full length. Ad12E1B enhanced transcriptional repression activity of hDaxx.
CONCLUSIONAd12E1B disrupts the co-localization of hDaxx with PML in PODs and enhances transcriptional repression activity of hDaxx.
Adaptor Proteins, Signal Transducing ; Adenovirus E1B Proteins ; physiology ; Carrier Proteins ; analysis ; genetics ; Cell Line, Tumor ; Humans ; Intracellular Signaling Peptides and Proteins ; Neoplasm Proteins ; analysis ; Nuclear Proteins ; analysis ; genetics ; Promyelocytic Leukemia Protein ; Repressor Proteins ; physiology ; Transcription Factors ; analysis ; Transcription, Genetic ; Tumor Suppressor Proteins
10.Epidemiological analysis of rabies in 2010, China.
Cui-Ping YIN ; Hang ZHOU ; Hui WU ; Xin-Xin SHEN ; Li-Hua WANG ; Wen-Wu YIN ; Shu-Mei WANG ; Qing TANG
Chinese Journal of Experimental and Clinical Virology 2011;25(6):434-436
OBJECTIVETo understand the related factors of rabies epidemic and provide the basic data for rabies control and prevention in China by statistic and retrospective analysis of rabies surveillance data in 2010.
METHODSWe used descriptive epidemiology method and statistic analysis to analyze the epidemiological characteristics of rabies in 2010 of China.
RESULTS2048 rabies cases were rabies cases were reported in 817 counties (districts) in 2010, which dropped 7.46% compares to 2009. The incidences in children and elder people were high; farmers are main occupation of the cases, the male to female ratio of the cases was 2.44:1. Children and older people are higher acquired rabies than other age population. 640 cases reported through national rabies sentinel surveillance system, 87.50% cases were caused by exposed to dogs, bite was the main exposure reason. The situation of deposing wounds was poor, and the use of vaccine was still low in individual cases, but in the rabies clinic cases under surveillance, the vaccine usage can reach 98%, the usage of immunoglobulin (RIG) or anti-serum for category III exposure in either group cases was not high.
CONCLUSIONThe epidemic of the rabies in 2010 was eased, Out-patient post-exposure prophylaxis was in good station, but there are still lots of problem existed: post-exposure prophylaxis of individual case was not desirable yet.
Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Rabies ; epidemiology ; prevention & control ; Time Factors