In order to improve the metabolism yield of cordycepin and adenosine,we studied nitrogen sources ,the levels of nitrogen sources and carbon sources,dynamic development in liquid culture ,as well as the total yield of cordycepin and adenosine in culture system through determining the contents of cordycepin and adenosine by HPLC .The results are as follows: not only animal protein but also some plant protein is very good nitrogen sources; in culture solution the suitable levels of nitrogen sources and carbon sources is respectively 3% and 4%; the content of adenosine in culture solution is very low, while it is quite high in mycelia; it is much higher that the total yield of cordycepin in culture solution than in mycelia; when the culture system is oscillated and cultured after 7~9 days, the total yield of cordycepin and adenosine are both high.

To obtain the key genes for chlorogenic acid biosynthesis of Lonicera hypoglauca, four new genes ware obtained from the our dataset of L. hypoglauca. And we also predicted the structure and function of LHPAL4, LHHCT1 , LHHCT2 and LHHCT3 proteins. The phylogenetic tree showed that LHPAL4 was closely related with LHPAL1, LHHCT1 was closely related with LHHCT3, LHHCT2 clustered into a single group. By Real-time PCR to detect the gene expressed level in different organs of L. hypoglauca, we found that the transcripted level of LHPAL4, LHHCT1 and LHHCT3 was the highest in defeat flowers, and the transcripted level of LHHCT2 was the highest in leaves. These result provided a basis to further analysis the mechanism of active ingredients in different organs, as well as the element for in vitro biosynthesis of active ingredients.
Chlorogenic Acid ; metabolism ; Cloning, Molecular ; Computational Biology ; Gene Expression Regulation, Plant ; Lonicera ; chemistry ; classification ; genetics ; metabolism ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Protein Structure, Secondary

Methyl valerate (MVA) pathway is one of the important ways for synthesis of terpenoids. This study was based on data of the transcriptome sequencing of Magnolia officinalis, the associated genes MoACOT, MoHMGS, MoHMGR, MoMK in methyl valerate (MVA) pathway, were completed in detail by using bioinformatics methods. The results of analysis showed that MoACOT and MoMK were stable hydrophobic proteins, MoHMGS and MoHMGR were unstable hydrophobic protein. The secondary structures of all proteins were hybrid architecture,and alpha helical were the major motifs. There were no clear transmembrane domains in MoACOT, MoHMGS and MoMK, but two transmembrane domains were founded in MoHMGR which were from 39-61 aa and 82-104 aa resepectively. The results of evolutionary relationship analysis showed that MoACOT, MoHMGS, MoHMGR and MoMK had relative close relationship to angiosperm or dicotyledonous plants, and accorded with genetic evolution rule. From transcriptome data, transcripted level of MoACOT, MoHMGS, MoHMGR, MoMK in M. officinalis and M. officinalis var. biloba was not significantly different. The result provided theoretical reference for study on Methyl valerate (MVA) pathway of terpenoid of M. officinalis.
Computational Biology ; Genes, Plant ; Magnolia ; genetics ; metabolism ; Phylogeny ; Terpenes ; metabolism

ObjectiveTo evaluate the effect of preoperative percutaneous transhepatic biliary drainage (PTBD) on postoperative outcomes after pancreaticoduodenectomy.MethodsThe clinical data of 115 patients undergoing pancreatoduodenectomy between 2001 and 2009 were retrospectively analyzed.The diagnosis of periampullary cancer or cancer of the pancreatic head was confirmed histologically.The preoperative total bilirubin level was more than 100 μmol/L and there was no concomitant cholangitis.Forty-two patients underwent PTBD (PTBD group),and 73 were not drained (early operation group).The following parameters were analyzed:wound infection,intra-abdominal abscess,intra-abdominal or gastrointestinal bleeding,biliary or pancreatic leakage,gastroparesis,morbidity and mortality.The length of hospital stay and cost were also assessed.ResultsThe perioperative mortality and morbidity were 2.38％/54.76％ in the PTBD group and 2.74％/50.68％ in the early operative group,respectively.There were no significantly differences between these two groups.Similar results were obtained in biliary leak,pancreatic leak,intra-abdominal infection,wound infection and gastroparesis.The length of hospital stay and cost were significantly less in the early operation group than the PTBD group.ConclusionsPTBD had no beneficial effects on postoperative outcomes following pancreaticoduodenectomy.For distal biliary obstruction,PTBD should not be carried out routinely.

Objective To study the relationship between anti-transcription intermediary factor 1 family proteins (TIF1) autoantibodies profiles and cancer-associated dermatomyositis (CAM) and to define the diagnostic value of different subtypes of anti-TIF1 aotuantibodies for CAM.Methods The sera from 156 patients with dermatomyositis (DM),55 with polymyositis (PM),70 with systemic lupus erythematosus (SLE),60 with rheumatoid arthritis (RA),46 with primary Sj(o)gren syndrome (pSS),14 with systemic sclerosis (SSc),49 with kinds of malignancies and 40 healthy subjects were examined by immunoprecipitation assays followed by western blotting.Statistical analysis were performed using ANOVA,t test Mann-Wittney U and x2 test or Fisher exact test,nonparametric method was used to evaluate the sensitivity and specificity through calculating the area under the receiver operating characteristic curve (ROC).Results In summary,32 of 156 sera from patients with DM (20.5％) were positive for at least one anti-TIF1 autoantibodies.There are four subtypes of anti-TIF1 autoantibodies profiles existed in patients with DM,including 4 patients with only positive anti-TIF1-α (12.5％),20 with only positive anti-TIF1-γ (62.5％),7 with both positive anti-TIF1-α and anti-TIF1-γ (21.9％) and 1 with both positive anti-TIF1-β and anti-TIF1-γ (3.1％).However,only positive anti-TIF1-α (7.3％) was observed in 4 patients with PM.No patients with other CTDs as well as malignancy and healthy subjects were positive for these autoantibodies.The sensitivity and specificity of presence of anti-TIF1-α antibodies for the diagnosis of CAM were 42.9％ and 96.5％,respectively and those of anti-TIF1-β antibodies were 0 and 99.3％,respectively and those of anti-TIF-1-γ antibodies were 64.3％ and 86.6％,respectively.Application of areas of ROC to identify the best performance of test of anti-TIF1 antibodies profiles were 0.70,0.50,0.76,0.74 and 0.71,respectively.Conclusion Joint detection of antiTIF1 autoantibodies profiles can improve the diagnostic capbility for CAM.

Cleistocalycis Operculati Cortex is the dry bark of Cleistocalyx operculatus. It is the raw material of Compound Hibiscuse which is external sterilization antipruritic drugs. The quality standard of Cleistocalycis Operculati Cortex in Guangdong Province "standard for the traditional Chinese medicine" (second volumes) only contains TLC identification. It is unable to effectively monitor and control the quality of Cleistocalycis Operculati Cortex. A reversed-phase HPLC method was established for the determination of 3, 3'-O-dimethylellagic acid from Cleistocalycis Operculati Cortex and the content was calculated by external standard method for the first time. Under the selected chromatographic conditions, the target components between peaks to achieve effective separation. 3,3'-O- dimethylellagic acid standard solution at the concentration of 1.00 - 25.0 mg x L(-1) showed a good linear relationship. The standard curve was Y = 77.33X + 7.904, r = 0.999 5. The average recovery was 101.0%, RSD was 1.3%. The HPLC method for the determination of 3,3'-O-dimethylellagic acid in Cleistocalycis Operculati Cortex is accurate and reliable. It can provide a strong technical support for monitoring the quality of Cleistocalycis Operculati Cortex.
Chromatography, High Pressure Liquid ; methods ; Chromatography, Reverse-Phase ; methods ; Drugs, Chinese Herbal ; analysis ; Plant Bark ; chemistry ; Syzygium ; chemistry

Objective Platelet activating factor(PAF),which has been implicated in the pathophysiology of inflammation in asthma,is degraded and inactivated by PAF acetlhydrolase(PAF AH).To investigate the association of PAF AH activity with genotype in asthmatic children.Methods We studied 57 asthmatic children and 30 normal controls. The plasma PAF AH genotype was detected as representative case with 3 different genotypes (Val/Val,Val/Phe and Phe/Phe) by allele specific polymerase chain reaction(AS PCR).The PAF AH activity in plasam was examined by the changes of substrate assay.Results In severe asthmatic individuals plasma PAF AH activities were lower than those of mild or moderate groups and control group,and plasma PAF AH activition was absent 15.4 %.In another three groups plasma PAF AH activation were absent 2 %-3 %.There was significant difference of plasma PAF AH activity among 3 groups of genotype(Val/Val,Val/Phe and Phe/Phe).In the similar genotype, there was no significant difference of plasma PAF AH activity between the groups of control and asthma.Conclusions There was imbalace of PAF/PAF AH in asthmatic children. In severe asthmatic individuals plasma PAF AH activities were lower than those of mild or moderate groups and control group. PAF AH(Val279Phe) gene mutation was related with plasma PAF AH activity.

OBJECTIVETo explore the effect and mechanism of hirudin on atherosclerotic plaques in apolipoprotein E knockout (ApoE(-/-)) mice.

METHODSTotally 24 ApoE(-/-) mice, 7-8 weeks old were fed with high fat diets. They were randomly divided into the recombinant hirudin treatment group (drug group) and the model group according to body weight and different dens, 12 in each group. Twelve C57BL/6J mice, 7-8 weeks old fed with high fat diet were recruited as the normal control group. Recombinant hirudin (0.25 mg/kg) was intraperitoneally injected to mice in the drug group from the 10th week old once every other day for five successive weeks. Equal volume of normal saline was injected to mice in the model group. Mice in the normal control group received no treatment. All mice were sacrificed after fed with high fat diet until they were 20 weeks old. Serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), high-density lipoprotein (HDL), high-sensitive C-reactive protein (hs-CRP), E-selectin, interleukin-6 (IL-6), and stromal metalloproteinase-2 (MMP-2) were detected. The plaque/lumen area and extracellular lipid composition/ plaque area were analyzed by HE staining and morphometry. Changes of signaling molecules in store-operated calcium channels, including stromal interacting molecule 1 (STIM1), Orail protein, and transient receptor potential channel 1 (TRPC1) were determined by Western blot. Results Lipid plaque formed in the aorta vessel wall of 20-week old mice in the model group. Compared with the normal control group, serum levels of TC, TG and LDL increased (P<0.01), hs-CRP, E-selction, IL-6, and MMP-2 obviously increased (P<0.01, P<0.05) in the model group; expression levels of STIM1, TRPC1, and Orail significantly increased (P<0.01). Compared with the model group, the plaque/lumen area and the extracellular lipid composition/plaque area significantly decreased in the drug group (P<0.05, P<0.01); serum levels of TC and LDL, hs-CRP, E-selction, IL-6, and MMP-2 obviously decreased (P<0.05, P<0.01); expression levels of STIM1, TRPC1, and Orail were significantly down-regulated (P<0.05, P<0.01).

CONCLUSIONHirudin could significantly improve lipids and endothelial functions of ApoE(-/-) mice, down-regulate expression levels of STIM1, Orai1, and TRPC1, and thus delaying the occurrence and development of atherosclerosis.

Animals ; Aorta ; Apolipoproteins E ; metabolism ; Atherosclerosis ; C-Reactive Protein ; Cholesterol ; Diet, High-Fat ; Drugs, Chinese Herbal ; E-Selectin ; Hirudins ; metabolism ; Interleukin-6 ; Lipids ; Lipoproteins, HDL ; Lipoproteins, LDL ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Plaque, Atherosclerotic ; metabolism ; Recombinant Proteins ; metabolism ; Triglycerides

Objective To study the value of transcatheter arterial chemoembolization (TACE)in combination with portal vein chemotherapy (PVC) after resection of hepatocellular carcinoma to prolong survival. Methods From January 2000 to July 2007, 168 patients with hepatocellular carcinoma (HCC) underwent tumor resection in our hospital. After operation, TACE in combination with PVC was performed in 48 patients (combined group), TACE alone in 26 (TACE group), PVC alone in 50 (PVC group) and none of the above in 44 (control group). All the patients were followed up for 17-96 months. The 1-, 3-and 5-year survival rates were compared among the 4 groups. Results Accumulative 1-, 3-and 5-year survival rates were higher in the combined and TACE groups than in PVC and control groups. Conclusion After resection of HCC, combined use of TACE and PVC is the same as TACE in prolonging patient survival. However, it is better than PVC alone and non-surgical procedure.

Epstein-Barr virus (EBV) is a human herpesvirus associated with important human diseases, including infectious mononucleosis syndrome, malignant lymphoma, and nasopharyngeal carcinoma. The mechanism of EBV entry into host cells remains a subject of intensive research. After decades of study, researchers have identified several key proteins and different patterns of EBV intrusion into host cells. The viral surface glycoproteins, gp350/220, gp42, gB, gH, and gL, are involved in interactions with the CR2 receptor on the surface of B lymphocytes during viral entry. However, the majority of epithelial cells lack CR2 receptor expression, which makes viral invasion much more complex than in B lymphocytes. Three different models have been proposed to explain how EBV enters epithelial cells: (1) "transfer of infection", mediated by B lymphocytes or Langerhans cells; (2) EBV utilizes its own proteins during the process of fusion with the cell membrane; and (3) progeny virions arising from EBV-infected epithelial cells cross lateral membranes into adjacent epithelial cells. This review will discuss the relevant mechanism of viral entry into B lymphocytes and epithelial cells during EBV infection.
Animals ; B-Lymphocytes ; virology ; Epithelial Cells ; virology ; Epstein-Barr Virus Infections ; virology ; Herpesvirus 4, Human ; genetics ; physiology ; Humans ; Viral Proteins ; genetics ; metabolism ; Virus Internalization