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

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

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.

The questionnaire and the result of 2010' Hwato Cup National University Students' Acupuncture Manipulation Skills Competition were analyzed in this paper. It was showed that the competition achieved the significant effects of enhancing acupuncture manipulation skills for the university students and promoting the standardization on teaching acupuncture manipulation skills. The teachers and students were not very satisfied with the current acupuncture manipulation skill education. In the competition, only the item of reciting the classics achieved more than 90 scores, which just displayed the memory ability. The results of the manipulation competition were generally not very high. It is suggested that concerning to teaching acupuncture and moxibustion in the future, the ratio of class time for improving the practice ability on the human body should be increased and the practice on some manipulation techniques such as inserting the needle by holding the needle tip with the pressing hand, reinforcing-reducing technique by twirling and rotating the needle and warming needle technique should be intensified. It is necessary to enhance the interpretation, emphasis and supplementation on the keys and details of some manipulations of acupuncture and moxibustion.
Acupuncture ; education ; Acupuncture Therapy ; instrumentation ; methods ; China ; Female ; Humans ; Male ; Surveys and Questionnaires ; Universities

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

OBJECTIVEBy reverse engineering and rapid prototyping techniques to found a new design method of maxillofacial restoration.

METHODSBy laser scanning apparatus the plaster face model was scanned and the primitive face point data were acquired. With the reverse engineering software, the point data were reconstructed to one smooth face surface image and the defect orbital tissue shape data was obtained from the normal contralateral tissue data in the software. The model designed the three-dimensional data of defect part and the rapid prototyping technique made the resin orbital restoration.

RESULTSThe laser scanning apparatus acquired the distinct and precise model data of the plaster face-model. The Digisurface retrograde engineer software succeeded to fulfill the unilateral orbital defect computer-aided design. The orbital restoration inosculated the plaster model tightly and symmetrically.

CONCLUSIONThe reverse engineering software and rapid prototyping technique could finish the computer-aided design and manufacture of the unilateral orbital defect restoration smoothly and satisfactorily.

Computer-Aided Design ; Humans ; Imaging, Three-Dimensional ; Orbit ; anatomy & histology ; injuries ; surgery ; Orbital Implants ; Prosthesis Design ; methods

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.