Isolation of high-quality genomic DNA from dried and processed crude drug is the key for the DNA identification of Dendrobii Caulis. The DNA extract of Dendrobii Caulis was firstly compared using different method to isolate genomic DNA from dried and processed crude drug, including commercial DNA extracted kit and CTAB method. Using modified CTAB method (extracted from large samples), the genomic DNA was successfully isolated from Dendrobii Caulis, including Huangcao and Fengdou. The ITS regions were amplified using the purified DNA as template, and then cloned and sequenced. These ITS sequences were compared with data from Genbank database and our lab, 14 Dendrobium species and five similar species were identified from "Huangcao" and "Huangcao" slice, while six species and three similar species from "Fengdou" according to their sequence similarity. The study demonstrated that the dried Dendrobii Caulis could be identified using DNA molecular method, which could overcome deficiencies and limitations of traditional identification method and has a certain application prospects.
DNA, Intergenic ; genetics ; DNA, Plant ; genetics ; Dendrobium ; classification ; genetics ; Sequence Analysis, DNA

Objective To observe pathological features of peripheral vessel injury caused by ex-plosion shock wave so as to provide theoretical basis for emergency treatment, prevention and complication reduction of war extremity injuries. Methods A total of 18 rabbits were randomly divided into three groups (six rabbits in each group) and placed respectively at 1, 2 and 3 m away from the explosion cen-ter. The animal model with blast injury was made by using fluid dynamite that electrically exploded at 60 cm above the ground. The physical parameters of blast wave were recorded using pressure transducers (PCB, UAS). After explosion, the femoral arteries were examined grossly, histologically and immunohis-tochemically. Results The results showed that vascular endothelial cells were denudated, the spaces of contractile fiber cells increased and appeared puff, the vassular elastic fibers ruptured, flexed and de-formed visibly. Some parts of the vessel wall ruptured completely or partly, leading to bleeding. TUNEL staining and fluorescence microscope found large number of apoptotic cells in endothelium layer, smooth muscle layer and membrana adventitia layer of the blood vessels. Conclusion Explosion shock wave can result in severe large blood vessel injury, which should be paid much attention during treatment of ex-plosion shock injury.

To explore a new method to identify Moutan Cortex to guarantee its safe use, internal transcribed spacer 2 (ITS2) sequence was used to identify Moutan Cortex and its adulterants. DNA was extracted and target fragments were amplified. Sequences were analyzed and assembled by CodonCode Aligner V3.7.1. Genetic distances were computed and phylogenetic tree was constructed based on kimura 2-parameter (K2P) model by MEGA 5.0. The length of the 20 ITS2 sequences of Moutan Cortex from nine different places is 227 bp, and no variation site was detected. The maximum inter-specificK2P distance of Moutan Cortex is 0, the minimum intra-specific K2P distance is 0.041, the average intra-specific K2P distance is 0.222. According to NJ analysis, Moutan Cortex from different places can get together as one branch with bootstrap support values 99%, which indicates Moutan Cortex can be easily distinguished from its adulterants. Using ITS2 sequence can accurately identify Moutan Cortex and its adulterants, it is an effective supplementary to traditional identification methods.
Base Sequence ; China ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Paeonia ; classification ; genetics ; Phylogeny ; Quality Control

To construct the Bac-to-Bac expression system of Bombyx mori nucleopolyhedrovirus (BmNPV), a transfer vector was constructed which contained an Escherichia coli (E. coli) mini-F replicon and a lacZ: attTN7: lacZ cassette within the upstream and downstream regions of the BmNPV polyhedrin gene. B. mori larvae were cotransfected with wild-type BmNPV genomic DNA and the transfer vector through subcutaneous injection to generate recombinant viruses by homologous recombination in vivo. The genomic DNA of budded viruses extracted from the hemolymph of the transfected larvae was used to transform E. coli DH10B. Recombinant bacmids were screened by kanamycin resistance, PCR and restriction enzyme (REN) digestion. One of the bacmid colonies, BmBacJS13, which had similar REN profiles to that of wild-type BmNPV, was selected for further research. To investigate the infectivity of BmBacJS13, the polyhedrin gene was introduced into the bacmid and the resultant recombinant (BmBacJS13-ph) was transfected to BmN cells. The budded viruses were collected from the supernatant of the transfected cells and used for infecting BmN cells. Growth curve analysis indicated that BmBacJS13-ph had a similar growth curve to that of wild-type BmNPV. Bio-assays indicated that BmBacJS13-ph was also infectious to B. mori larvae.

Objective To observe the clinical curative effect of Annaohuoxue capsules, a prescription of promoting blood circulation and removing blood stasis, on patients with hypertensive cerebral hemorrhage and explore its safety. Methods Ninety-three patients with hypertensive cerebral hemorrhage,admitted to our hospital from June 2010 to November 201 i,were chosen and randomly divided into Arnaohuoxue capsule treatment group (group A,n=31),Tiandantongluo capsule treatment group (group B,n=32) and placebo control group (group C,n=30); After 7 d of onset,treatments were given,respectively,for 21 d.Hemorrheology,scores of neurological impairment,Glasgow coma scale (GCS) scores were recorded and compared in these groups before and after treatment. Results The plasma viscosity,and aggregation index of R BCs in patients from group A were significantly lower than those in patients from group B (P＜0.05); statistically significant difference on the clinical efficacy was noted between each 2 groups (P＜0.05):the outcome of group A was the best,that of group B was better,and that of group C was the worst.The GCS scores in patients from group A were significantly higher than those in patients from group B and group C (P＜0.05) Conclusion Annaohuoxue capsules can improve the hemorrheology indices and neurological impairment,and decrease the GCS scores in varying degrees,and the treatment efficiency of Annaohuoxue capsules is much better than placebo.

OBJECTIVETO establish a RP-HPLC method for the determination of calycosin-7-O-beta-D-glucopyranoside in Radix Astragali, and to analyse the calycosin-7-O-beta-D-glucopyranoside content of ten samples of Radix Astragali, collected from different regions.

METHODA Polaris C18(250 mm x 4.6 mm, 5 microns) column was used and a mixture of methanol-water (30:70) was used as the mobile phase at a flow rate of 1.0 mL.min-1. The column temperature was 25 degrees C and the UV detection wavelength was 254 nm.

RESULTThe calibration curve was in good linearity over the range of 0.0106-2.12 micrograms with the regression equation Y = 3035. 97 X - 14.85(r = 0.9999). The average recovery was 95.8% (n = 5, RSD = 1.3%).

CONCLUSIONThe method is simple, quick, sensitive and reproducible. In all of the samples, the calycosin-7-O-beta-D-glucopyranoside contents differ markedly.

Astragalus membranaceus ; chemistry ; classification ; China ; Chromatography, High Pressure Liquid ; Ecosystem ; Glucosides ; analysis ; Isoflavones ; analysis ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Species Specificity

AIMTo study the effect of Angelica sinsensis polysaccharides on lymphocyte proliferation and induction of IFN-gamma.

METHODSAngelica sinensis polysaccharides(AP) were separated into AP-I, AP-II, AP-III and AP-IV by alcohol deposition with different concentration. The radioactivities of [3H]-TdR uptake by lymphocyte were used to determine the ability of lymphocyte. The bioactivity of IFN-gamma was measured by violet crystalline dying.

RESULTSAP-IV was found to be composed of Ara and Glu in the ratio of 0.99:6.47, the molecular weight was estimated to be 5,600. AP-I and AP-II 100 mg.kg-1 i.p. were found to significantly augment mice splenocyte proliferation, release IFN-gamma and increase IFN-gamma bioactivity. 50 micrograms.mL-1 AP-I, AP-II and AP-III were shown to enhance the proliferative response of the mouse spleen lymphocytes in vitro.

CONCLUSIONAP-I and AP-II showed higher immunoactivity than AP-III, AP-IV had no effect.

Angelica sinensis ; chemistry ; Animals ; Cell Division ; drug effects ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Immunity ; drug effects ; Interferon-gamma ; biosynthesis ; Lymphocytes ; cytology ; drug effects ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Plants, Medicinal ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology ; Random Allocation ; Spleen ; cytology ; Tumor Cells, Cultured

Estrogen participates in many life activities through combination with estrogen receptor alpha (ERalpha) or estrogen receptor beta (ERbeta) in the body. In order to establish an in vitro estrogen-like compound screening model, the coding region of human ERalpha and ERbeta was separately constructed into pET32-ERalpha and pET43-ERbeta prokaryotic expression vector and water-soluble recombinant ERalpha and ERbeta proteins were expressed in Escherichia coli strain BL21. Western blotting revealed that both recombinant proteins have estrogen receptor binding sites. The proteins were purified using S-Tag affinity Purification Kit and digested with enterokinase to get the ERalpha and ERbeta proteins. About 0.90 mg of ERalpha and 0.65 mg of ERbeta were obtained at the concentration of 0.181 and 0.131 mg x mL(-1), respectively.
Binding Sites ; Escherichia coli ; metabolism ; Estrogen Receptor alpha ; genetics ; metabolism ; Estrogen Receptor beta ; genetics ; metabolism ; Genetic Vectors ; Humans ; Protein Binding ; Recombinant Proteins ; genetics ; metabolism

OBJECTIVETo develop an HPLC method for the determination of acteoside in Radix Rehmanniae.

METHODThe chromatographic conditions were as follows: Polaris C18(4.6 mm x 250 mm, 5 microm) column, a mobile phase in gradient mode composed of acetonitrile 0.1% acetic acid solution, a flow rate of 1.0 mL x min(-1), and 334 nm as the detection wavelength.

RESULTActeoside showed good linear relationship at the range of 10-500 microg x mL(-1) (r = 0.9990). The average recovery was 100.1%, RS D 3.7%.

CONCLUSIONThe proposed method promised to be applicable for the quality control of Radix Rehmanniae.

Chromatography, High Pressure Liquid ; methods ; Glucosides ; analysis ; Phenols ; analysis ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Rehmannia ; chemistry ; Reproducibility of Results ; Sensitivity and Specificity

To investigate the hematological abnormality and clinical characteristics in systemic lupus erythematosus (SLE), the hematological data of 58 SLE and the curative effects of corticosteroid and immunosuppressive agents on SLE were retrospectively analysed by using SPSS/PC software. The results showed that the incidence of hematological abnormalities in 58 cases was as follows: 50 cases of hemogram abnormality (86.2%), 41 of anemia (70.7%), 34 of thrombocytopenia (58.7%), 37 of leukopenia (63.8%). Peripheral cytopenia of every cell lineage was common in SLE. The cell abnormalities of two or three lineages were seen in 41 cases (70.7%). The initial symptoms with hematological abnormality were found in 12 cases (20.7%), 7 out of 12 cases were erroneously diagnosed as hematology diseases (12.1%). In 30 out of 58 patients, the results of bone marrow examination showed that 23 had hyperplasia (76.7%) and 7 were hypoplasia. In 25 out of 38 cases, splenomegaly (65.8%) was found by B ultrasonography. In 25 patients with SLE receiving Coombs test, 3 were positive (12.0%). PAIg increased in 16 out of 22 cases of thrombocytopenia (72.7%). 26 cases of SLE with two or three lineage cytopenia in peripheral blood were treated by corticosteroid and immunosuppressive agent. The hemogram improved in all patients including 6 cases of bone marrow hypoplasia. It is concluded that the hematological abnormalities are frequent in SLE patients, which are short of specialty. The cytopenia of two or more lineage in peripheral blood is most common when bone marrow shows hyperplastic. The therapy with corticosteroid and immunosuppressive agents is efficacious.
Adolescent ; Adult ; Aged ; Bone Marrow Examination ; Child ; Female ; Hematologic Diseases ; etiology ; Humans ; Lupus Erythematosus, Systemic ; blood ; complications ; therapy ; Male ; Middle Aged