The study is aimed to ensure the quality and safety of medicinal plants by using ITS2 DNA barcode technology to identify Corydalis boweri, Meconopsis horridula and their close related species. The DNA of 13 herb samples including C. boweri and M. horridula from Lhasa of Tibet was extracted, ITS PCR were amplified and sequenced. Both assembled and web downloaded 71 ITS2 sequences were removed of 5. 8S and 28S. Multiple sequence alignment was completed and the intraspecific and interspecific genetic distances were calculated by MEGA 5.0, while the neighbor-joining phylogenetic trees were constructed. We also predicted the ITS2 secondary structure of C. boweri, M. horridula and their close related species. The results showed that ITS2 as DNA barcode was able to identify C. boweri, M. horridula as well as well as their close related species effectively. The established based on ITS2 barcode method provides the regular and safe detection technology for identification of C. boweri, M. horridula and their close related species, adulterants and counterfeits, in order to ensure their quality control, safe medication, reasonable development and utilization.
Base Sequence ; China ; Corydalis ; chemistry ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; chemistry ; genetics ; DNA, Ribosomal Spacer ; chemistry ; genetics ; Molecular Sequence Data ; Nucleic Acid Conformation ; Papaveraceae ; chemistry ; classification ; genetics ; Phylogeny ; Plants, Medicinal ; chemistry ; classification ; genetics

OBJECTIVEThe survival rate of cadaveric renal transplant in children has been improved following the development of transplantation technology and the application of immunosuppressive agents. In this study, the prognosis of renal transplantation, operative procedure and immunosuppressive agents administration in 21 children with end-stage renal disease (ESRD) were analyzed.

METHODSFrom January 1985 to December 2001, 21 patients (9 males and 12 females with a mean age of 14 +/- 2 yr, mean body weight of 33.4 kg and mean height of 136.5 cm) received renal transplantation because of ESRD were enrolled in the study. The patients with an average GFR of 8.28 ml/min were managed with dialysis for 13.4 months in average pro-transplantation. All cadaveric kidneys were from adults, which included 1 donor with one HLA mismatch, 3 with two mismatches, 5 with three mismatches and 4 with four mismatches. All transplantations were performed with anastomoses of the adults' renal artery and vein to the children's iliac externa artery and iliac externa vein. Biological inducement therapy was given in 4 cases. At the first 3 - 5 days post-transplantation, methylprednisolone was administered [7 mg/(kg.d)]. All patients received baseline diploid or triple immunosuppression therapy of cyclosporin A [6 - 8 mg/(kg.d)] or FK506 [0.18 - 0.25 mg/(kg.d)], mycophenolate mofetil [MMF 10 - 15 mg/(kg.d)] or azathioprine [1 - 3 mg/(kg.d)] and prednisone [0.4 - 0.6 mg/(kg.d)]. High-dose methylprednisolone [10 mg/(kg.d)] was administered to control the acute rejection.

RESULTSThe renal function of patients was restored 5.6 days in average after transplantations. The 1st, 3rd and 5th year survival rates of recipient/graft were 95.2%/95.2%, 86.7%/73.3% and 72.7%/63.6%, respectively. One case had super-acute renal rejection, 5 cases had acute rejection, 3 cases had delayed graft function and 3 patients died. The longest survival time was 12 years. The major complications included hypertension (47.6%), diabetes (19.4%), infection (19.4%) and drug-induced hepatic injury (14.2%). Catch-up growth was seen in most of the pediatric recipients.

CONCLUSIONRenal transplantation is the most ideal method to treat children with ESRD, and the growth of the pediatric patients will be improved after transplantation. Adult donor kidneys adapt to the school age patient. And the protocol of immunosuppressive therapy (prednisone plus MMF and FK506) should be applied.

Adolescent ; Adult ; Child ; Female ; Histocompatibility Testing ; Humans ; Kidney Failure, Chronic ; mortality ; therapy ; Kidney Transplantation ; Male ; Postoperative Care ; Preoperative Care ; Prognosis ; Survival Rate ; Time Factors

OBJECTIVEto investigate the clinical characteristics in two families with early repolarization syndrome (ERS) and recurrent syncope.

METHODall family members including the probands were screened with routine clinical examination, electrocardiography, echocardiography, Holter recording, chest x-ray, head-up tilt test and blood biochemistry.

RESULTSthere was no clinical evidence of organic heart disease in all members from the two families. Proband 1 showed recurrent syncope, ERS and repeated torsade de pointes ventricular tachycardia and ventricular fibrillation were documented with resting ECG. ERS was detected in one brother, one nephew and one son from him and all were free of cardiac events including syncope, cardiac arrest and sudden cardiac death. Proband 2 showed recurrent syncope, ERS and ST segment arched upward elevation in V(1)-V(3) were documented by ECG. His father suffered sudden cardiac death at the age of 65 and asymptomatic ERS was detected in one of his nephew.

CONCLUSIONSERS is not always linked with benign clinical course and can sometimes lead to repeated syncope, torsade de pointes ventricular tachycardia and ventricular fibrillation. Pedigree research is of importance for ERS.

Adult ; Arrhythmias, Cardiac ; genetics ; Asian Continental Ancestry Group ; Humans ; Male ; Pedigree ; Recurrence ; Syncope ; genetics ; physiopathology ; Syndrome

OBJECTIVEIn order to characterize the feature of stress response induced by stressor with both physical and psychological natures, the effects of water restriction performed in different experimental modes on the physiological parameters, psychological behavioral manifestations and brain c-Fos expressions were observed and compared.

METHODSFifty-eight male Wistar rats were used and randomly divided into three experimental groups (n = 18 for each) and a control group (n = 4). In control group, the rats were allowed to access drinking water freely at all experimental period. In the experimental groups the water supply to the rats was restricted. In timed water supply (TW) group, the water was supplied twice a day, 10 min for each in fixed hours every day. In empty bottle-served (EB) and water-restricted (WR) groups, the water was served only once a day for 10 min, either in the early morning or evening, and in the other time point scheduled for water supply only an empty bottle without water was provided in the EB group and nothing was given in the WR group. The quantities of drank water and eaten food, weight-gaining, and behavior score were observed every day. The serum level of corticosterone was assayed and the rats were sacrificed with fixative perfusion of 3 d, 7 d or 14 d, respectively, following water restriction (n = 6 for each time point in each group). The brain c-Fos expressions were examined with immunohistochemical method.

RESULTSThe slow down of weight-gaining, rise of serum corticosterone level, occurrence of psychological behavioral manifestations of unpeaceful restlessness such as exploring and attacking, enhance of c-Fos expression in the subfornical organ (SFO), median preoptic nucleus (MnPO), area postrema (AP), hypothalamic paraventricular nucleus (PVN), supraoptic nucleus (SON), medial (MeA) and central (CeA) amygdaloid nucleus and ventrolateral septum (LSV) were noticed in both EB and WR groups, except the nucleus of solitary tract (NTS) in which the Fos expression was decreased. The changes of Fos expression in most of nuclei in EB group began at day 3, at least persisted till day 7, and backed down at day 14, while in WR group, similar changes started at day 7 and reached its peak at day 14. In TW group, only the concentration of corticosterone at day 7 was slightly increased and the rest indexes observed were unchanged.

CONCLUSIONThe results indicate that water restriction induces physical as well as psychological stress responses. And the water restrictions experimentally executed in different modes result in different manifestations of behavioral response and brain immediately early gene expression in discrete brain nuclei/regions.

The MDMV (Maize Dwarf Mosaic Virus, MDMV) CP (Coat Protein, CP) gene was cloned by RT-PCR method and introduced into the embryonic calli derived from immature embryos of elite inbred 18-599hong and 18-599bai via particle bombardment. Bombarded calli were selected on selection medium containing 5-10 mg/L (PPT) Bialaphos. From resistant calli, 79 plantlets were regenerated. 18 of 79 were grown and harvested. The results of Southern blotting and PCR analysis demonstrated that MDMV CP have been integrated into the genome of the transgenic plants. PCR-positive progeny plants were artificially inoculated with MDMV strain B, and the average chlorosis of the functional leaves of each plant was investigated. The typical symptoms were observed from the leaves of the control inbreds. while, the presence of the MDMV CP gene provided resistance to inoculation with MDMV strain B.
Capsid Proteins ; genetics ; Cloning, Molecular ; Mosaic Viruses ; genetics ; Plant Diseases ; genetics ; virology ; Plants, Genetically Modified ; Transfection ; Zea mays ; genetics ; virology

OBJECTIVETo investigate and compare the effect of radio-frequency (RF) field exposure on expression of heat shock proteins (Hsps) in three human glioma cell lines (MO54, A172, and T98).

METHODSCells were exposed to sham or 1950 MHz continuous-wave for 1 h. Specific absorption rates (SARs) were 1 and 10 W/kg. Localization and expression of Hsp27 and phosphorylated Hsp27 ((78) Ser) (p-Hsp27) were examined by immunocytochemistry. Expression levels of Hsp27, p-Hs27, and Hsp70 were determined by Western blotting.

RESULTSThe Hsp27 was primarily located within the cytoplasm, p-Hsp27 in both cytoplasm and nuclei of MO54, A172, and T98 cells. RF field exposure did not affect the distribution or expression of Hsp27. In addition, Western blotting showed no significant differences in protein expression of Hsp27 or Hsp70 between sham- and RF field-exposed cells at a SAR of 1 W/kg and 10 W/kg for 1 h in three cells lines. Exposure to RF field at a SAR of 10 W/kg for 1 h slightly decreased the protein level of phosphorylated Hsp27 in MO54 cells.

CONCLUSIONThe 1950 MHz RF field has only little or no apparent effect on Hsp70 and Hsp27 expression in MO54, A172, and T98 cells.

Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; radiation effects ; Glioma ; Heat-Shock Proteins ; genetics ; metabolism ; Humans ; Neoplasm Proteins ; genetics ; metabolism ; Neuroglia ; radiation effects ; Protein Transport

OBJECTIVETo observe the effect of electromagnetic pulse (EMP) exposure on cerebral micro vascular permeability in rats.

METHODSThe whole-body of male Sprague-Dawley rats were exposed or sham exposed to 200 pulses or 400 pulses (1 Hz) of EMP at 200 kV/m. At 0.5, 1, 3, 6, and 12 h after EMP exposure, the permeability of cerebral micro vascular was detected by transmission electron microscopy and immunohistochemistry using lanthanum nitrate and endogenous albumin as vascular tracers, respectively.

RESULTSThe lanthanum nitrate tracer was limited to the micro vascular lumen with no lanthanum nitrate or albumin tracer extravasation in control rat brain. After EMP exposure, the lanthanum nitrate ions reached the tight junction, basal lamina and pericapillary tissue. Similarly, the albumin immunopositive staining was identified in pericapillary tissue. The changes in brain micro vascular permeability were transient, the leakage of micro vascular vessels appeared at 1 h, and reached its peak at 3 h, and nearly recovered at 12 h, after EMP exposure. In addition, the leakage of micro vascular was more obvious after exposure of EMP at 400 pulses than after exposure of EMP at 200 pulses.

CONCLUSIONExposure to 200 and 400 pulses (1 Hz) of EMP at 200 kV/m can increase cerebral micro vascular permeability in rats, which is recoverable.

Animals ; Brain ; blood supply ; Capillary Permeability ; physiology ; Electromagnetic Fields ; adverse effects ; Electrophysiology ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the distribution of hepatitis C virus (HCV) genotypes in Yixing, Jiangsu province.

METHODSGenotypes identification on sera samples were obtained from 158 donors who had already been anti-HCV positive through PCR method with type specific primer designed according to the sequence of 5'non-coding region (5'NCR). 5'NCR was also sequenced and compared with published date. Genotypes distribution was investigated in patients with different sex and clinical types of hepatitis C.

RESULTSOf the total 158 patients, 95 were HCV RNA positive in which 80 patients having genotype 1b (80/95; 84.4%), 5 patients having genotype 2(5/95; 5.3%), 5 patients with 1b/2 mixed genotypes (5/ 95; 5.3%) and another 5 patients whose genotype undetermined. The difference on the distribution of HCV genotypes was significant between female and male patients (P < 0.05) but not in different kinds of hepatitis C patients.

CONCLUSIONType 1b was the predominant HCV genotype in Yixing area.

Base Sequence ; Blood Donors ; China ; epidemiology ; Female ; Genotype ; Hepacivirus ; genetics ; isolation & purification ; Hepatitis C ; epidemiology ; prevention & control ; therapy ; virology ; Humans ; Male ; Middle Aged ; Sequence Analysis, DNA ; Sex Factors

This study was aimed to establish ELISA for recombinant bovine IFN-gamma (BovIFN-gamma) detection and provide a new method for diagnosis of pathogenic infection. The total RNA was isolated from peripheral blood leucocytes cultured with PHA mitogen stimulation. Then bovine IFN-gamma (BovIFN-gamma) gene cDNA was amplified by RT-PCR and cloned into pET28a to obtain the expression plasmid designated as pETBovIFN-gamma. The pETBovlFN-gamma was further transformed into competent E. coli BL21 cells and a 18kD His-tagged protein as expected was expressed after IPTG induction. By using purified recombinant BovIFN-gamma as antigen and lymphocyte-hybridoma technique, four hybridoma cell lines which stably secreted monoclonal antibodies against rBovIFN-gamma were generated, designated as A7, A10, G6, and G10. The immunoglobin subset was identified as IgG1 . Western-blotting analysis and ELISA demonstrated that the monoclonal antibodies secreted by all the four hybridoma cell lines could react specifically to the recombinant BovIFN-gamma, but not irrelative proteins such as Ag85B, ESAT-6-CFP-10 and GM-CSF, suggesting that the four hybridoma cell lines were rBovIFN-gamma specific monoclonal antibodies. A sandwich ELISA was established by using A10 secreted monoclonal antibody and rabbit polyclonal antibodies against BovIFN-gamma, HRP labeled goat anti-rabbit IgG. The results indicated that the sensitivity was 2ng/mL. This sandwich ELISA to detect BovIFN-gamma paved the way to develop a sensitive method for specific infection detection such as bovine tuberculosis diagnosis.
Animals ; Animals, Newborn ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibody Specificity ; immunology ; Blotting, Western ; Cattle ; Cells, Cultured ; DNA, Complementary ; genetics ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Hybridomas ; Interferon-gamma ; genetics ; immunology ; metabolism ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; Rabbits ; Recombinant Proteins ; immunology ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

To research the influence of Reduning injection on the activity and mRNA expression of cytochrome P450 (CYP450) system in rat liver microsomes. Rat liver microsomes were prepared after a seven-days continuous administration of Reduning injection. An HPLC-MS method was applied to determine the specific metabolites of CYP450 probe substrates in rat liver microsomal incubations. The activity of CYP450 isozymes were represented by the formation of metabolites. The Real-time quantitative polymerase chain reaction (Q-PCR) was applied to determine the mRNA expression levels of CYP450. Reduning injection significantly reduced the activity of CYP2B1, 2C12, 2C13 (P < 0.01), but did not affect CYPlA2; low dose and high dose of Reduning injection had an inhibition trend on the activity of CYP2D2, but did not statistically differ from control group; low dose of Reduning injection significantly induced the activity of CYP3A1 (P < 0.01), high dose of Reduning injection had an induce trend on the activity of CYP3A1, but did not statistically differ from control. At the mRNA level, low and high dose of Reduning injection had an induce trend on the expression of CYP1A2, 2C11, 2D1, 2E1, 3A1, but did not statistically differ from control. Reduning injection significantly induced the activity of CYP2B1. Reduning injection significantly induced the activity of CYP3A1 in mRNA expression and enzyme activity levels, which may result adverse drug reaction after being combined with macrolides antibiotics. Reduning injection significantly reduced the activity of CYP2B1, 2C12, 2C13, 2D2 in enzyme activity levels, when combined with other drugs, it should be fully taken into account of the possible drug-drug interaction in order to avoid adverse side effects.
Animals ; Cytochrome P-450 Enzyme System ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Injections ; Isoenzymes ; metabolism ; Male ; Microsomes, Liver ; drug effects ; enzymology ; Rats ; Rats, Sprague-Dawley