OBJECTIVE: To identify the characteristics of recombinant adenovirus modified PBMC-derived dendritic cells to resist the HIV-1 infection. METHODS: The recombinant adenovirus vector was integrated with CCR5A32, CCR5siRNA, HIV-1 poli and HIV-1 inti genes by using AdEasy system. The human PBMC from healthy donor blood was isolated in order to get dendritic cells. The expression of CCR5Δ32, CCR5, CXCR4 and HIV-1 P24 in PBMC or modified cells was measured by Westernblot. RESULTS: After the cells have been modified by Ad-R5Δ32siHIV, the expression of CCR5Δ32 was increased while the expression of CCR5 and CXCR4 were decreased. The level of p24 was lower in modified cells than that of unmodified cells. The modified cells show resistance to HIV-1 infection. CONCLUSION: The recombinant adenovirus-modified cells show good resistance to HIV-1 infection. Modification of HSC-derived immunity cells, such as DCs, may be a potent strategy to resist HIV-1 infection.

Objective: With the aim to obtain crocetin glycosyltransferase UGTCs4 in cultured saffron suspension cell, and carry out its bioinformatics and expression mode analysis. Methods: A homologous cloning strategy and 5’ RACE methods were adopted, the full-length cDNA sequence of a crocetin glycosyltransferase, designated UGTCs4 (GeneBank number: KX398932), was obtained. The characteristics of physiochemical properties, structure and function of the deduced UGTCs4 protein were determined using a series of bioinformatics tools. Semi-quantitative PCR was used for gene expression analysis. Results: The results showed that the full length cDNA of UGTCs4 was 1 380 bp in length and encoding a 459 amino acid; UGTCs4 had high identities (83.2%) with UGTCs2 protein from saffron; UTGTCs4 had the same evolutionary tree as UGTCs2. UGTCs4 transcripts were constitutively expressed in the leaves, stems, and roots. UGTCs4 gene could respond to multiple treatments of indoleacetic acid (IAA), abscisic acid (ABA), gibberellin (GA), hydrogen peroxide (H2O2), and methyl jasmonate (MJA), which promoted its transcription. Conclusion: cDNA of crocetin glycosyltransferase was cloned from suspension cells for the first time and the response of UGTCs4 to different inducers was confirmed. Molecular characterization of UGTCs4 will be useful for further functional determination of the gene involving in the crocin biosynthesis and expression regulation.

Teeth are the hardest organs in the human body. They are not easily affected by external physical and chemical factors to degrade or deform. Dental age assessment has been widely used in forensic practice. Dental image is one of the most common methods in the age estimation. The emergence of cone beam computed tomography (CBCT) technology provides a new way to obtain three-dimensional image of teeth. It has many advantages such as low-dose radiation, short-time scanning, high-precision image, and finical convenience. Recent development of CBCT technology and its application on age estimation are reviewed in this paper.
Age Determination by Teeth/methods* ; Cone-Beam Computed Tomography/trends* ; Humans ; Imaging, Three-Dimensional

OBJECTIVETo isolate the active component from Dioscorea cirrhosa Lour and test its activity in lowering blood pressure.

METHODSThe serial components were obtained from the total extract, ligroin extract, ethyl acetate, n-butyl alcohol extract and water extract. The isolated active components were administered in rats via the common carotid artery canulation and tail vein injection to test their effects on blood pressure.

RESULTSThe component A isolated and purified from normal butanol showed obvious effect in lowering the blood pressure of rats.

CONCLUSIONThe isolated active component from Dioscorea cirrhosa Lour possesses obvious blood pressure-lowering activity.

Animals ; Antihypertensive Agents ; isolation & purification ; pharmacology ; Blood Pressure ; drug effects ; Dioscorea ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Male ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley

This study investigated the effect of arctigenin (Arc) on the cell activation, cytokines expression, proliferation, and cell-cycle distribution of mouse T lymphocytes. Mouse lymphocytes were prepared from lymph node and treated with Phorbol-12-myristate-13-acetate (PMA)/Ionimycin (Ion) and/or Arc. CD69, CD25, cytokines, proliferation and cell cycle were assayed by flow cytometry. The results showed that, at concentrations of less than 1.00 micromol x L(-1), Arc expressed non-obvious cell damage to cultured lymphocytes, however, it could significantly down-regulate the expression of CD69 and CD25, as well as TNF-alpha, IFN-gamma, IL-2, IL-4, IL-6 and IL-10 on PMA/Ion stimulated lymphocytes. At the same time, Arc could also inhibit the proliferation of PMA/Ion-activated lymphocytes and exhibited lymphocyte G 0/G1 phase cycle arrest. These results suggest that Arc possesses significant anti-inflammatory effects that may be mediated through the regulation of cell activation, cytokines expression and cell proliferation.
Animals ; Anti-Inflammatory Agents ; isolation & purification ; pharmacology ; Antigens, CD ; metabolism ; Antigens, Differentiation, T-Lymphocyte ; metabolism ; Arctium ; chemistry ; Cell Cycle Checkpoints ; drug effects ; Cell Proliferation ; drug effects ; Cytokines ; metabolism ; Female ; Furans ; isolation & purification ; pharmacology ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-2 ; metabolism ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-6 ; metabolism ; Ionomycin ; pharmacology ; Lectins, C-Type ; metabolism ; Lignans ; isolation & purification ; pharmacology ; Lymphocyte Activation ; drug effects ; Mice ; Mice, Inbred BALB C ; Plants, Medicinal ; chemistry ; T-Lymphocytes ; cytology ; drug effects ; immunology ; Tetradecanoylphorbol Acetate ; pharmacology ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo study the incidence of injuries among residents living in the communities in Zhejiang.

METHODSA household questionnaire survey was conducted to 16 899 residents in four communities.

RESULTSThe incidence of injuries was 16.11% in total, higher in urban area than in rural area and higher in men than in women. The leading causes of injuries were falls (4.80%), knife-cutting or by sharp articles (3.86%), traffic accident (2.07%), collision (1.99%), scalds (1.13%) and animal bites (1.06%). Peak incidence of age group was different in different injuries, such as traffic accident was high in 45 - 60 age group and falls was high in 5 and 60 age group. Direct economic loss for each case with injury would cost 324.21 RMB yuan.

CONCLUSIONInjury was common and frequently occurred among residents at the community level which seriously on danger their health and families.

Accidental Falls ; statistics & numerical data ; Accidents, Traffic ; statistics & numerical data ; Adolescent ; Adult ; Aged ; Animals ; Bites and Stings ; epidemiology ; Burns ; epidemiology ; Child ; China ; epidemiology ; Community Health Services ; Dogs ; Female ; Humans ; Male ; Middle Aged ; Prevalence ; Wounds and Injuries ; epidemiology ; mortality

OBJECTIVETo develop an objective bioassay for quantitative detection of HIV-induced cell-cell fusion for screening HIV entry inhibitors.

METHODSHL2/3 cells expressing HIV envelope proteins gp120/gp41, Tat, and other HIV proteins were co-cultured with HeLa-CD4-LTR-beta-gal cells expressing CD4 receptor and HIV LTR triggered reporter gene beta-galactosidase. The enzyme activities of beta-galactosidase were detected by a chromogenic substrate, chlorophenol red-beta-galactopyranoside (CPRG). Specific HIV entry inhibitors were used to validate the established detecting method.

RESULTSNo syncytium was formed by mixing HL2/3 and HeLa-CD4-LTR-beta-gal cells. However, the membrane could be fused and the Tat expressed by HL2/3 cells could bind to HIV LTR on HeLa-CD4-LTR-beta-gal cells and trigger the expression of beta-galactosidase. CPRG allowed quantitative and sensitive detection of the activity of beta-galactosidase. Further studies showed that HIV entry inhibitors could inhibit the activity of beta-galactosidase in a dose-dependent manner.

CONCLUSIONWe have developed a simple, cheap, objective and quantitative non-infectious cell-cell fusion bioassay that can be used to screen for anti-HIV agents targeting the virus entry from natural and synthetic compound libraries.

Biological Assay ; Cell Fusion ; Cell Line ; Coculture Techniques ; Drug Evaluation, Preclinical ; methods ; HIV Envelope Protein gp120 ; metabolism ; HIV Envelope Protein gp41 ; metabolism ; HIV Fusion Inhibitors ; chemistry ; pharmacology ; Humans ; beta-Galactosidase ; metabolism

BACKGROUNDParaquat (PQ; 1, 1'-dimethyl-4, 4'-bipyridinium), a widely used herbicide that is structurally similar to the known dopaminergic neurotoxicant MPTP (1-methyl-1, 2, 3, 6-tetrahydropyridine), has been suggested as a potential etiologic factor for the development of Parkinson's disease (PD). Aging is an accepted risk factor for idiopathic Parkinson's disease. The aim of this study was to test the hypothesis that paraquat could induce PD-like nigrostriatal dopaminergic degeneration in aging C57BL/6 mice.

METHODSSenile male C57BL/6 mice were intraperitoneally injected with either saline or PQ at 2-day intervals for a total of 10 doses. Locomotor activity and performance on the pole test were measured 7 days after the last injection and animals were sacrificed one day later. Level of dopamine (DA) and its metabolites levels in the striatum were measured by high-performance liquid chromatography with an electrochemical detector (HPLC-ECD), and numbers of tyrosine hydroxylase (TH) positive neurons were estimated using immunohistochemistry.

RESULTSLocomotor activities were significantly decreased and the behavioral performance on the pole test were significantly impaired in the PQ treated group. Level of DA and its metabolites levels in the striatum were declined by 8 days after the last injection. Immunohistochemical analyses showed that PQ was associated with a reduction in numbers of tyrosine hydroxylase positive neurons.

CONCLUSIONSLong-term repeated exposes to PQ can selectively impair the nigrostriatal dopaminergic system of senile mice, suggesting that PQ could play an important role in the pathogenesis of Parkinson's disease (PD). Our results also validate a novel model of PD induced by exposure to a toxic environmental agent.

Aging ; pathology ; Animals ; Corpus Striatum ; drug effects ; Disease Models, Animal ; Dopamine ; analysis ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Motor Activity ; drug effects ; Paraquat ; toxicity ; Parkinson Disease, Secondary ; chemically induced ; Substantia Nigra ; drug effects ; enzymology ; Tyrosine 3-Monooxygenase ; analysis

Ultrafine powder and cell wall-broken powder of herbal medicine lack of the morphological characters and microscopic identification features. This makes it hard to identify herb's authenticity with traditional methods. We tested ITS2 sequence as DNA barcode in identification of herbal medicine in ultrafine powder and cell wall-broken powder in this study. We extracted genomic DNAs of 93 samples of 31 representative herbal medicines (28 species), which include whole plant, roots and bulbs, stems, leaves, flowers, fruits and seeds. The ITS2 sequences were amplified and sequenced bidirectionally. The ITS2 sequences were identified using Basic Local Alignment Search Tool (BLAST) method in the GenBank database and DNA barcoding system to identify the herbal medicine. The genetic distance was analyzed using the Kimura 2-parameter (K2P) model and the Neighbor-joining (NJ) phylogenetic tree was constructed using MEGA 6.0. The results showed that DNA can be extracted successfully from 93 samples and high quality ITS2 sequences can be amplified. All 31 herbal medicines can get correct identification via BLAST method. The ITS2 sequences of raw material medicines, ultrafine powder and cell wall-broken powder have same sequence in 26 herbal medicines, while the ITS2 sequences in other 5 herbal medicines exhibited variation. The maximum intraspecific genetic-distances of each species were all less than the minimum interspecific genetic distances. ITS2 sequences of each species are all converged to their standard DNA barcodes using NJ method. Therefore, using ITS2 barcode can accurately and effectively distinguish ultrafine powder and cell wall-broken powder of herbal medicine. It provides a new molecular method to identify ultrafine powder and cell wall-broken powder of herbal medicine in the quality control and market supervision.
Cell Wall ; DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drugs, Chinese Herbal ; analysis ; Phylogeny ; Plants, Medicinal ; classification ; genetics ; Powders ; Quality Control

OBJECTIVETo review the results for minimally invasive aortic valve replacement (AVR) through a 5 cm right anterolateral thoracotomy.

METHODSFrom July 2009 to September 2011, 101 consecutive patients with isolated aortic valve disease (degenerative in 37 patients, rheumatic in 21 patients, congenital in 37 patients, endocarditic in 3 patients and aorta-arteritis in 1 patients) underwent AVR through the right anterolateral thoracotomy approach in the third intercostal space with a groin incision for femoral connection of cardiopulmonary bypass. The mean age was 45.7 years (ranging from 17 to 71 years). Sixty patients were male.

RESULTSOperations were successfully performed in all but 1 patient (1.0%) who required intraoperative conversion to full sternotomy. Mean duration of cardiopulmonary bypass time and aortic cross-clamp time was (88 ± 24) minutes and (55 ± 18) minutes, respectively. Thirty-day mortality was 1.0% (1/101), this patient was found difficult in weaning off cardiopulmonary bypass and exhibited severe coronary artery plaque, although bypass graft was carried out immediately, the patient died of severe low cardiac output syndrome finally. No blood products were needed in 83.2% patients. Follow-up was performed in all patients at an average of (16 ± 7) months postoperatively. A good recovery was obtained in all patients except one who died of multiple organ failure caused by massive cerebral infarction 38 days after surgery.

CONCLUSIONSMinimally invasive aortic valve replacement though the right anterolateral thoracotomy approach is safe and feasible, with good cosmetic results and rapid postoperative recovery. It is worthy of clinical elective application.

Adolescent ; Adult ; Aged ; Aortic Valve ; surgery ; Female ; Heart Defects, Congenital ; surgery ; Heart Valve Diseases ; surgery ; Heart Valve Prosthesis Implantation ; methods ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Treatment Outcome ; Young Adult