The total triterpene saponins of Psammosilene tunicoides have significant pharmacologic activity. Psammosilene tunicoides squalene synthase (PSS) is a gateway enzyme to regulate the biosynthesis of total triterpene saponins extracted from the root of Psammosilene tunicoides which is an endangered species. In this paper, cDNA encoding of PSS was cloned by the degenerate primer PCR and rapid-amplification of cDNA ends (RACE). The full-length of cDNA of PSS is 1663 bp, with an open reading frame (ORF) of 1 245 bp, encoding 414 amino acid polypeptide (calculated molecular mass, 47.69 kDa), 5'UTR (untranslated region) and 3'UTR are 260 bp and 158 bp, respectively. The deduced amino acid sequence of PSS has higher homology with the known squalene synthases of several species such as Panax notoginseng (83%), Panax ginseng (82%) and Glycyrrhiza glabra (82%) than that with Schizosacharomyces pombe (35%), Candida albicans (39%) and Homo sapiens (47%). The characterization of PSS was done by a series of methods, such as prokaryotic expression, the activity of enzyme in vitro, capillary gas chromatography (GC) and capillary gas chromatography mass spectrometry (GC-MS). The results showed that the cell-free extract of E. coli transformed with the recombinant plasmid can effectively convert farnesyl diphosphate into squalene in vitro. GenBank accession number is EF585250. Our research provided important base for the study of Psammosilene tunicoides secondary metabolism and metabolic engineering.
Caryophyllaceae ; enzymology ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Endangered Species ; Escherichia coli ; genetics ; metabolism ; Farnesyl-Diphosphate Farnesyltransferase ; genetics ; metabolism ; Gas Chromatography-Mass Spectrometry ; Open Reading Frames ; Phylogeny ; Plant Proteins ; genetics ; metabolism ; Plants, Medicinal ; chemistry ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; metabolism ; Sequence Homology, Amino Acid ; Transformation, Genetic

Objective To explore the biological functions of retinoic acid-inducible gene-I(RIG-I) in vivo through phenotype analysis of RIG-I knockout mice. Methods The gene expression of RIG-Ⅰ in various tissues of mice was examined with Northern blotting and semi-quantitative RT-PCR.The phenotypes observed included body weight measurement,differential count of peripheral blood cells,metabolic parameters measurement and histopathologic examination. ResultsRIG-Ⅰ expressed in various tissues of mice with different levels.No gross developmental abnormalities and expected maturation arrest in granulocytic differentiation were observed in RIG-Ⅰ knockout mice.However,RIG-Ⅰ knockout mice exhibited an unexpected increase in the ratios of neutrophiles to lymphocytes in peripheral blood and increased susceptibility to bacteria infection. Conclusion RIG-Ⅰ may play an important role in immune regulation in mice.

A L9 (3(4)) orthogonal design table to be used to get nine combinations of extraction of three herbs of Wuji pill: Coptis chinensis, Tetradium ruticarpum and Paeonia lactiflora Pall., and nine extraction of single herbs correspondingly, altogether eighteen combinations. Quantification of five representative bioactive ingredients: berberine, palmatine, evodiamine, rutaecarpine, paeoniflorin in rat liver by ultra high liquid chromatography-tandem mass spectrometry after oral administration at 2 h time point of eighteen combinations. The result shows the bioactive ingredients have different concentrations betweem different combinations and the single herb with the same dosage significantly as well as the same dose combinations. C. chinensis with evodiamine concentration of low and high dose T. ruticarpum was positively correlated. T. ruticarpum with berberine concentration of low dose C. chinensis was negatively correlated and of meddle dose C. chinensis was correlated positively. T. ruticarpum with paeoniflorin concentration of middle dose P. lactiflora was correlated positively. P. lactiflora with palmatine concentration of middle dose C. chinensis was negatively correlated and with evodiamine and rutaecarpine concentration of middle dose T. ruticarpum was negatively correlated. These shows the three single herbs interactions resulted in the differences of each ingredients concentration in rat liver. The orthogonal analysis indicates the combination 12: 6: 6 make the maximum concentration in rat liver.
Administration, Oral ; Animals ; Biological Availability ; Biomedical Research ; methods ; Chromatography, High Pressure Liquid ; methods ; Drug Stability ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Liver ; metabolism ; Male ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; Temperature

We investigated the causative agents of hand, foot and mouth disease (HFMD) in children in Beijing epidemic from April to June of 2007. Totally, 82 specimens (including throat swabs, rectal swabs and vesicular swabs) were collected from 51 patients with HFMD. All patients had typical skin lesions, but no neurological manifestations. These clinical specimens were directly tested by RT-PCR assay with three primer sets: universal enterovirus, CA16-specific and EV71-specific primers. Enterovirus, CA16 and EV71 were identified depending on the size of PCR products. The percentage of the specimens identified as positive for enterovirus was 70.6%. Of the 51 cases, 25 were positive for CA16, 4 were positive for EV71, and 7 were enterovirus positive of non-CA16 or EV71. The ratio among them was about 6:1:2. EV71 strains belonged to genotype C4 after sequencing and phylogenetic analysis.
Child, Preschool ; China ; Enterovirus ; classification ; genetics ; isolation & purification ; physiology ; Female ; Genotype ; Hand, Foot and Mouth Disease ; virology ; Humans ; Male ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo evaluate the practical value and the advantages of globe geographical positioning with wireless communication monitoring and supervision system in the field survey.

METHODSSpots which were randomly sampled by the National Ministry of Health for the investigation were chosen in the endemic areas for schistosomiasis in Jiangsu, Jiangxi, Anhui, Hunan, Hubei, Sichuan and Yunnan provinces. Portable GPS CEC9680 was used for collecting relevant waypoints and track, recording on-the-spot geographical positions. The positioning data package was sent back synchronously in the form of short message of SMS to the monitoring service center, and the moving routes of the terminal receiver monitored were displayed on the GIS map to achieve real-time supervision and staff scheduling.

RESULTSWith globe geographical positioning with wireless communication monitoring and supervision system, accurate positioning of 12 spots in the provinces of Jiangsu and 3 trial spots for schistosomiasis control with comprehensive treatment designated by the State Council has been established with real-time communicating recording, and monitoring systems.

CONCLUSIONThe globe geographical positioning with wireless communication monitoring and supervision system has provided a technical platform for the survey of schistosomiasis and other infectious diseases.

Animals ; China ; Environmental Monitoring ; methods ; Epidemiological Monitoring ; Geographic Information Systems ; Humans ; Schistosomiasis ; epidemiology ; prevention & control ; Schistosomiasis japonica ; prevention & control

Objective：This study was designed to explore the mechanism of inhibition of proliferation induced by arsenic trioxide (As2O3) and homoharringtonine in neuroblastoma. Method：Methy thiazoloyl tetrazolium (MTT) methods and analysis of apoptosis were used to evaluate the effect of As2O3 and homoharringtonine on the proliferation of NB cell lines and their mechanisms. Reverse-transcriptase polymerase chain reaction(RT-PCR) and immunohistochemistry techniques were used in NB cell lines treated with As2O3 and HHT to determine N-myc mRNA and protein expression. Results：Both As2O3 and HHT can significantly inhibit the proliferation of neuroblastoma cell lines and induce apoptosis of SJ-N-SH and IMR-32 cells. In As2O3 treated cell line, at transcription and protein level, N-myc expression is up-regulated at first and then down-regulated to a lower level. HHT can down-regulate N-myc protein. Conclusion：N-myc may play some roles in apoptosis induced by As2O3 and HHT, but the mechanisms initiating N-myc gene may be different.

OBJECTIVETo evaluate the effect of alprazolam use on psychological status and hospitalization cost in patient with paroxysmal supraventricular tachycardia underwent electrophysiology studies or radiofrequency catheter ablation.

METHODSIn this prospective, randomized, double-blind, placebo-controlled study, 142 inpatients [77 males, mean age (43.1 +/- 14.5) years] were randomly assigned to receive alprazolam (0.4 mg qd at 10PM for 3 days, n = 72) or placebo (n = 70) 3 days before scheduled electrophysiology studies or radiofrequency catheter ablation. All patients were examined by the Chinese version of Symptom Checklist-90 (SCL-90) at 24 hours before the procedure.

RESULTSCompared with the placebo group, the scores of somatization (1.38 +/- 0.40 vs. 1.65 +/- 0.56, P < 0.01), anxiety (1.50 +/- 0.39 vs. 1.69 +/- 0.50, P < 0.05), phobic anxiety (1.24 +/- 0.36 vs. 1.47 +/- 0.57, P < 0.01), psychotism constructs (1.24 +/- 0.34 vs. 1.35 +/- 0.30, P < 0.05) and global severity index (1.36 +/- 0.35 vs. 1.49 +/- 0.37, P < 0.05) were significantly decreased in alprazolam group. The hospitalization costs were also significantly lower in alprazolam group (32 498 +/- 1170) yuan compared to placebo group (32 947 +/- 1096) yuan, P < 0.05.

CONCLUSIONThe alprazolam use before electrophysiology studies and radiofrequency catheter ablation can improve the patients' psychological status and reduce the hospitalization costs.

Adolescent ; Adult ; Aged ; Alprazolam ; therapeutic use ; Anti-Anxiety Agents ; therapeutic use ; Catheter Ablation ; economics ; psychology ; Double-Blind Method ; Female ; Hospitalization ; economics ; Humans ; Male ; Middle Aged ; Prospective Studies ; Tachycardia, Paroxysmal ; psychology ; therapy ; Tachycardia, Supraventricular ; psychology ; therapy ; Young Adult

OBJECTIVETo examine the differences in the expressions of c-kit, HIWI and vimentin in the testis and epididymis of rats aged different months, investigate the regular changes in the expression of characteristic molecules in the ripening and senescent process of the testis and epididymis, and provide experimental evidence for studies on the aging of the male reproductive system.

METHODSSprague-Dawley male rats were divided into a young group (6-month-old, n=10), an adult group (12-month-old, n=10) and an aged group (24-month-old, n=10). The immunohistochemical SP method was used to examine c-kit, HIWI and vimentin in the testis and epididymis of the rats.

RESULTSThe positive immunohistochemical reaction to c-kit was observed mainly in spermatogonia, weakly in other cells and also in the epithelia and lumens of the epididymis. The abundant expression of HIWI was detected in all stages of spermatogenic cells in the testis and epididymis of the 6-month-old and 12-month-old rats. Moreover, spermatogonias and primary spermatocytes displayed intense expression. Sertoli cells, Leydig cells, myoid cells and vascular endothelial cells (VEC) were also HIWI-positive cells. But the positive expression decreased with age in the testis and epididymis of the 24-month-old rats (P > 0.05). vimentin expression was weak in the testis and epididymis of the 6-month-olds but increased significantly in the 24-month-olds (P > 0.01).

CONCLUSIONThe expressions of c-kit and HIWI decreased in the testis and epididymis of the 24-month-old rats, while vimentin expression increased markedly with age. The results suggest that the aging of the testis and epididymis is closely related to the abnormal transduction of cell signals.

Age Factors ; Animals ; Epididymis ; metabolism ; Immunohistochemistry ; Male ; Proteins ; metabolism ; Proto-Oncogene Proteins c-kit ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Testis ; metabolism ; Vimentin ; biosynthesis

OBJECTIVESTo investigate possible abnormalities of the sperm neck in eight infertile painters observed by transmission electron microscope.

METHODSSpermatozoa from 8 infertile painters (21-32 years old) were observed by transmission electron microscope.

RESULTSThe abnormalities of the connecting piece could be divided into 6 types, among which the anomaly of the connecting piece concomitant with a true nuclemalamic condition was the most serious.

CONCLUSIONSThe abnormalities may be related to expression obstacles of nuclear genome and mitochondrial genome, and to anomalous cellular signal transduction during spermiogenesis as well.

Adult ; Humans ; Infertility, Male ; pathology ; Male ; Microscopy, Electron, Transmission ; Occupational Exposure ; Paint ; adverse effects ; Spermatozoa ; ultrastructure

OBJECTIVETo investigate the expression of miR-9 in B lymphocytes, B cell lymphoma and classical Hodgkin's lymphoma (cHL) cell lines and its significance.

METHODSCD19(+) B lymphocytes were sorted from normal lymph node by magnetic beads. Total cellular micro-RNA was extracted from cHL cell line L428, B cell lymphoma cell lines Ly1 and Ly10 (diffuse large B cell lymphoma), Raji cells (Burkitt's lymphoma) and CD19(+) B lymphocytes, respectively. These micro-RNAs were separately transformed into cDNA by reverse transcription. The expression levels of miR-9 were measured by fluorescence quantitative PCR. In situ hybridization was used to detect the expression of miR-9 in cell lines.

RESULTSThe expression of miR-9 was high in L428 cells (104.44 ± 1.61), and low in cell lines of B cell lymphoma (Ly1: 2.17 ± 0.38; Ly10: 1 ± 0.015; Raji: 2.65 ± 0.89), and extremely low in CD19(+) B lymphocytes (0.0026 ± 0.00040). Compared with that in the other cell lines, the expression of miR-9 in L428 cells was statistically significant (P < 0.05). miR-9 localized in the cytoplasm diffusely and strongly in L428, but scattered and slightly with some prominent distribution around the nuclear membranes in Ly1 and Ly10, and only weakly in Raji.

CONCLUSIONSmiR-9 highly expressed in cHL cell line and might be a molecular marker for diagnosis and treatment of cHL.

B-Lymphocytes ; metabolism ; Cell Line, Tumor ; Cell Lineage ; Hodgkin Disease ; metabolism ; pathology ; Humans ; Lymphoma, B-Cell ; metabolism ; pathology ; MicroRNAs ; metabolism