According to the transcriptome dataset of Panax notoginseng, the key geranylgeranyl pyrophosphate synthase gene (GGPPS) in terpenoid backbone biosynthesis was selected to be cloned. Using specific primer pairs combining with RACE (rapid amplification of cDNA ends) technique, the full-length cDNA sequence with 1 203 bp, which containing a 1 035 bp open reading frame, was cloned and named as PnGGPPS. The corresponding full-length DNA sequence contained 2 370 bp, consisted of 1 intron and 2 exons. The deduced protein PnGGPPS contained 344 amino acids and shared more than 73% identity with GGPPS from Ricinus communis and Salvia miltiorrhiza. PnGGPPS also had specific Aspartic acid enrichment regions and other conserved domains, which belonged to the Isoprenoid-Biosyn-C1 superfamily. The quantitative real-time PCR showed that PnGGPPS expressed in different tissues of 1, 2, 3 years old root, stem, leaf and 3 years old flower, and the expression level in 3 years old leaf was significant higher than that in other organs, which suggested that it might not only be involved in the regulation of the growth and development, but also be associated with the biosynthesis of chlorophyll and carotenoids, the development of chloroplast, the shade habit and the quality formation of P. notoginseng.
Cloning, Molecular ; Computational Biology ; Geranylgeranyl-Diphosphate Geranylgeranyltransferase ; genetics ; Panax notoginseng ; genetics ; Real-Time Polymerase Chain Reaction

To investigate the morphogenetic process of human adenovirus type 41 (HAdV-41), 293TE cells were infected with purified wild-type HAdV-41, and ultrathin sections of infected cells were prepared and observed under a transmission electron microscope. Results showed that HAdV-41 entered host cells mainly through three ways: non-clathrin-coated pit, clathrin-coated pit, and direct penetration of plasma membrane. In addition, cell microvilli might help HAdV-41 enter cells. After entering into cells, HAdV-41 virus particles could be found in vacuoles or lysosomes or be in a free state in cytoplasm. Only free virus particles could be found near nuclear pores (NP), suggesting that the virus needed to escape from lysosomes for effective infection and viral nucleoprotein entered the nucleus through NP. Progeny viruses were as-sembled in the nucleus. Three types of inclusion bodies, which were termed as fibrillous inclusion body, condense inclusion body, and stripped condense inclusion body, were involved in HAdV-41 morphogenesis. In the late phase of viral replication, the membrane integrity of the infected cells was lost and viral particles were released extracellularly. This study reveals the partial process of HAdV-41 morphogenesis and provides more biological information on HAdV-41.
Adenovirus Infections, Human ; virology ; Adenoviruses, Human ; genetics ; growth & development ; physiology ; ultrastructure ; Cell Membrane ; virology ; Cell Nucleus ; virology ; Humans ; Virus Release ; Virus Replication

Moderate drought stress has been found to promote the accumulation of active ingredients in Glycyrrhiza uralensis root and hence improve the medicinal quality. In this study, the transcriptomes of 6-month-old moderate drought stressed and control G. uralensis root (the relative water content in soil was 40%-45% and 70%-75%, respectively) were sequenced using Illumina HiSeq 2000. A total of 80,490 490 and 82 588 278 clean reads, 94,828 and 305,100 unigenes with N50 sequence of 1,007 and 1,125 nt were obtained in drought treated and control transcriptome, respectively. Differentially expressed genes analysis revealed that the genes of some cell wall enzymes such as β-xylosidase, legumain and GDP-L-fucose synthase were down-regulated indicating that moderate drought stress might inhibit the primary cell wall degradation and programmed cell death in root cells. The genes of some key enzymes involved in terpenoid and flavonoid biosynthesis were up-regulated by moderate drought stress might be the reason for the enhancement for the active ingredients accumulation in G. uralensis root. The promotion of the biosynthesis and signal transduction of auxin, ethylene and cytokinins by moderate drought stress might enhance the root formation and cell proliferation. The promotion of the biosynthesis and signal transduction of abscisic acid and jasmonic acid by moderate drought stress might enhance the drought stress tolerance in G. uralensis. The inhibition of the biosynthesis and signal transduction of gibberellin and brassinolide by moderate drought stress might retard the shoot growth in G. uralensis.
Droughts ; Gene Expression Regulation, Plant ; Glycyrrhiza uralensis ; genetics ; Plant Roots ; Sequence Analysis, DNA ; Stress, Physiological ; Transcriptome

OBJECTIVETo analyze the therapeutic effect of human neural precursor cells transplantation in treatment of neonates with severe brain injury.

METHODThe transplantation was performed on 6 newborns, one of them was diagnosed as extremely severe carbon monoxide poisoning at 5(th) day after birth; one of them was diagnosed as severe hypoglycemia; the others had asphyxia at birth with Apgar scores from 1 to 3 and were diagnosed as severe neonatal asphyxia, severe hypoxic ischemic encephalopathy according to images, electroencephalogram, biochemical examination and clinical manifestation. With the approval of hospital ethics committee and informed consent of the family members, the newborns received human neural precursor cells transplantation at the 4(th) to 20(th) day after birth. With the agreement of a pregnant woman, forebrain cells were obtained from the forebrain of her 12-week old fetus after spontaneous abortion. The cells from the fetal brain were amplified into human neural precursor cells in vitro and were injected into the cerebral ventricle of the patients.

RESULTOn the 2(nd) day after transplantation, sucking and swallowing reflexes gradually appeared in all the patients, muscular tension was also improved, and convulsion stopped. NBNA scoring in 3 of the patients reached normal level on the 28(th) day after birth. The 6 patients were followed up for 12 months. Four patients were normal in psychomotor development and scores of each scale reached normal level. Two patients have cerebral palsy.

CONCLUSIONhNPCs transplantation is safe and effective in treatment of severe neonatal brain injury. More clinical trials and further observation are needed.

Brain Injuries ; surgery ; Female ; Humans ; Hypoxia-Ischemia, Brain ; surgery ; Infant, Newborn ; Male ; Neural Stem Cells ; cytology ; transplantation

AIM To establish an HPLC method for the simultaneous content determination of gallic acid,protocatechuic acid,morroniside,loganin,sweroside,paeoniflorin,hypericin,astragalin,salvianolic acid B,salvianolic acid A,epimedin C and icariin in Bushen Huoxue Sanjie Capsules.METHODS The analysis was performed on a 30℃thermostatic Agilent 5 TC-C18 column(250 mm×4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1%phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 240 nm.RESULTS Twelve constituents showed good linear relationships within their own ranges(r≥0.999 8),whose average recoveries were 97.11%-101.14%with the RSDs of 0.60%-2.65%.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of Bushen Huoxue Sanjie Capsules.

AIM To study the chemical constituents of the secondary metabolites of endophytic fungi DL02 from Saposhnikovia divaricata (Turcz.) Schischk.and their biological activities.METHODS The ethyl acetate and methanol extract leavening from S.divaricata was isolated and purified by silica and Sephadex LH-20,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The antifungal and antibacterial activities were determined by the minimum inhibitory concentration method.RESULTS Seven compounds were isolated and identified as β-sitosterol (1),kojic acid (2),stigmasta-7,22-diene-3β,5α,6α-triol (3),uracil (4),allantoin (5),erythritol (6) and adenosine (7).Compound 4 had strong inhibitory effects on pseudomonas aeruginosa with the MIC values of 31.3 μg/mL.CONCLUSION Compounds 2-5 and 7 are isolated from this fungus for the first time.Compounds 4-5 have strong biological activities.

Meridians and Acupoints Science is a basic course and important component of acupuncture and Tuina science. This course contains considerable theoretical and basic knowledge to be memorized, which causes some difficulties for students to study and inevitably influences the learning effect. In view of that, the authors implemented reforms in interest teaching practice in teaching of Meridians and Acupoints Science. Reforms including the use of interesting memory method, string comparison of acupoints, interspersed discussion of cases, presentation of famous doctors' experience in acupoint application and development of the second class activities have achieved some success. That has aroused the students' enthusiasm for learning and improved the quality of teaching.

Objective To evaluate the therapeutic effects of poly(N-isopropylacrylamide)adriamycin magnetic nanoparticles(ADM-PNIPAM-Fe_3O_4)on liver VX2 tumor in rabbits via transcatheter arterial chemoembolization.Methods VX2 tumor pieces were successfully implanted into liver lobes of rabbits with liver tumors formation.All the animals were randomly divided into 4 groups of 8 each.Group A(control group)injected with 10 ml physiologic saline,Group B(ADM group)treated with free adriamycin (1 mg/kg)via arterial infusion,Group C(ADM-PNIPAM group)ADM-PNIPAM(1.5 mg/kg)was infused through arterial route,Group D(ADM-PNIPAM-Fe_3O_4+M group)ADM-PNIPAM-Fe_3O_4(2 mg/kg)was infused through arterial route;all were in individual doses,respectively;meanwhile an persistant magnet with intensity of 0.4 T was stabilized at the tumor region.Spiral CT scans were performed to measure size of liver tumors and evaluate lung metastasis at 1 day before operation and 14 days after operation.All experimental animals were sacrficed on the 15th days after operation and followed by pathologic and histologic examination of the tumor and lung specimens including changes befor and after the operation with correlative comparisons.Results There were no significant difference in volumes of tumors among 4 groups at 1 day before operation.The average tumor volume in the group A was(23.87?7.02)cm~3 at 14 days after operation;(7.70?1.53)cm3 in group B;(4.29?0.25)cm~3 in group C;(2.05?0.18)cm~3 in group D. The average tumor volumes in the group B,group C and group D were significantly smaller than that in the control group A at same time after operation and there was significant difference among the three experimental groups.According to the order of tumor sizes from small to large was as follows:group D＜group C＜group B＜group A.It showed that the average size of group D(ADM-PNIPAM-Fe_3O_4+M group) was the smallest among the 4 groups.Lung metastatic rates were 100%,66.7%,37.5% and 12.5% in group A,group B,group C and group D at 14 days after operation,respectively.Lung metastatic rates of group D was lower than that of group A,but there were no significant differences among group A,B and C.But pathological examination showed that there were lesser number of lung metastatic tumors in group B and C than that of group A .The intratumoral necrotic degrees showed as group D＞group C＞group B＞group A. Conclusions ADM-PNIPAM-Fe_3O_4 treatment for liver tumor via vascular interventional method combined with magnetic field localized in the lesion possesses significant inhibitory effect on tumor growth of liver VX2 tumor in rabbits.ADM-PNIPAM-Fe_3O_4 is thus initially confirmed as a kind of effective praeputium in interventional chemoembolization.

Objective To investigate the treatment status of antiparkinsonism in Xi'an. Methods Six general hospitals were randomly chosen in Xi' an and all Parkinson' s disease (PD) patients were interviewed by questionnaire from Jan. 2007 to Apr. 2007. Results 92 PD outpatients were enrolled in, including 48 males and 44 females, from 43 to 86 years old (mean 65.6±17.1) with duration of the disease from 0.2 to 27.8 years (mean 4.4 ±9.4). The preference of the drug use from the patients were: 40 (43.5%) preferred taking levodopa, 25 (27.2%) with amantadine and/or trihexyphenidyl, 14(15.2%) with levodopa and others, 4(4.4%) with dopamine agonist and others, 2 (2.2%) with other drugs, 7 (7.6%) with no treatment. There were 69 (75.0%) patients onset with resting tremor, 15 (16.3%) with bradykinesia, 6 (6.5%) with rigidity, and 2 (2.2%) with unknown symptoms. There was no startically significant difference in anti-PD drugs among the patients onset with different symptoms (P>0.05). 45 patients appeared the onset of disease before 65 years old and with no dementia, 47 onset after 65 with or without dementia. There was no significant difference of anti-PD drugs between the two groups (P>0.05). Most patients initiated anti-PD treatment with levodopa but few of them chose dopamine agonist. According to the classification of Hoehn & Yahr, 25(27.2%) belonged to Grade Ⅰ, 53(57.6%) to Grade Ⅱ ,8(8.7%) to Grade Ⅲ ,3(3.3%) to Grade Ⅳ and 3 (3.3%) to Grade Ⅴ. There was no significant differences of anti-PD drugs between different grades of the disease (P>0.05). 55.3% of the patients changed their anti-PD drugs randomly during the therapy, but with no relation to their gender, age, educational level, dementia, the number of family members, course of diseases, or the degree of Hoehn & Yahr, frequency and categories of medicine. Conclusion Anti-PD treatment in Xi' an did not strictly follow the standardized protocol, with few patients using dopamine agonist and over 50% of the patients changed their drugs randomly.

OBJECTIVETo prepare a lymph node-targeted ultrasound/fluorescence bi-functional imaging contrast agents, and observe its effectiveness both on contrast-enhanced ultrasound (CEUS) and vivo near infrared fluorescence (NIR) imaging through animal experiments.

METHODSThe chimeric lymph node-targeted ligand (phosphatidylserine) and near-infrared fluorescent substance were assembled to form bi-functional contrast microbubbles. The morphology and size distribution were detected by optical microscope and Malvern potential tests. Five normal New Zealand white rabbits were subcutaneously injected with the prepared contrast agent in bilateral footpads, and the imaging effectiveness of lymph nodes and lymphatic vessel were observed by CEUS and NIR technique. Then blue dye was subcutaneously injected at the same site, and the rabbits were sacrificed for lymph nodes pathological examination.

RESULTSLipid ultrasound microbubbles,with a mean size of 3-5 Μm in diameter, appeared to be uniform in distribution and regular in configuration. The images of inflow lymphatic vessel and relevant lymph node were quickly showed up after the subcutaneous injection by CEUS, which was identical to the result detected by NIR. Biopsy confirmed that all the blue-stained lymph nodes could be displayed by NIR.

CONCLUSIONSThe self-made bi-functional contrast agent has a good imaging ability in CEUS and NIR imaging. It may be a better agent as lymph node tracer.

Animals ; Contrast Media ; chemistry ; Fluoresceins ; chemistry ; Lymph Nodes ; anatomy & histology ; diagnostic imaging ; Lymphatic Metastasis ; pathology ; Male ; Rabbits ; Ultrasonography