1.Pathological study on peripheral vessel injury induced by explosion shock wave
Jianmin LIU ; Zhi YUAN ; Jian LIU ; Xuefei ZHANG ; Rong Lü ; Long BI ; Yunyu HU
Chinese Journal of Trauma 2009;25(8):751-754
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.
2.Identification of Dendrobii Caulis basing on ITS sequence.
Zi YE ; Ye LU ; Zheng-Tao WANG ; Hong XU ; Zhi-Bi HU
China Journal of Chinese Materia Medica 2014;39(20):3928-3935
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
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genetics
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DNA, Plant
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genetics
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Dendrobium
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classification
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genetics
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Sequence Analysis, DNA
3.Construction of the Bac-to-Bac System of Bombyx mori Nucleopolyhedroviru
Jin-shan, HUANG ; Bi-fang, HAO ; Xiu-lian, SUN ; Fei, DENG ; Hua-lin, WANG ; Zhi-hong, HU
Virologica Sinica 2007;22(3):218-225
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.
4.Identification of moutan cortex and its adulterants by ITS2 sequence.
Meng WEI ; Lan WU ; Yuan TU ; Wei-Chao REN ; Li XIANG ; Wei SUN ; Lin-Bi ZHANG ; Zhi-Gang HU
China Journal of Chinese Materia Medica 2014;39(12):2180-2183
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
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China
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DNA Barcoding, Taxonomic
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methods
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DNA, Plant
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genetics
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DNA, Ribosomal Spacer
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genetics
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Drug Contamination
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prevention & control
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Drugs, Chinese Herbal
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chemistry
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classification
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Molecular Sequence Data
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Paeonia
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classification
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genetics
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Phylogeny
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Quality Control
5.Clinical curative effect of Annaohuoxue capsules, a prescription of promoting blood circulation and removing blood stasis, on patients with hypertensive cerebral hemorrhage
Zhi-Hu YU ; Hua XIAO ; Zhen-Lin ZHAO ; Dong HU ; Qing-Hua ZHANG ; Zhong-Sheng BI
Chinese Journal of Neuromedicine 2012;11(6):622-625
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.
6.Effect of Angelica sinensis polysaccharides on lymphocyte proliferation and induction of IFN-gamma.
Jun-jie SHAN ; Yi WANG ; Shun-chun WANG ; Di LIU ; Zhi-bi HU
Acta Pharmaceutica Sinica 2002;37(7):497-500
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
7.Effect of alexandrite laser treatment for hair removal in Tibet mini-pigs.
Dong ZENG ; Wen-lin YU ; Yuan BI ; Chuan-hong YANG ; Huang-wen LAI ; Zhi-qi HU
Journal of Southern Medical University 2009;29(4):697-700
OBJECTIVETo observe the histological and ultrastructural changes of the skin and hair follicles following hair removal by alexandrite laser in Tibet mini-pigs.
METHODSTwelve healthy Tibet mini-pigs with dark hair were treated with alexandrite laser for hair removal. The skin specimens were taken immediately and at 1 h and 1, 3, 5, 10, 15, 30, 60 days after the laser treatment for observation under optical and transmission electron microscope.
RESULTSLaser hair removal resulted in extensive coagulation necrosis, carbonization and falling of the subcutaneous hair shafts, and some of the cells in the outer root sheath and hair bulb underwent degenerative and necrotic changes. One hour after laser treatment, the cells in the outer root sheath and bulb exhibited nuclear condensation, fragmentation and or karyolysis characteristic of cell apoptosis. The cell apoptosis reached the peak level on day 3 after the laser exposure, accompanied by endothelial degeneration in the hair papilla vessels, edema and lymphocyte infiltration in the dermal tissues. Tissue reaction and inflammation were relieved on day 5, and the dermal tissue and follicles recovered their normal structures on day 10. At 60 days after the treatment, the hair follicles decreased markedly but the structure of the residue follicles remained normal.
CONCLUSIONAlexandrite laser exposure results in selective destruction of the follicles by inducing direct coagulation and cell apoptosis to achieve permanent hair removal. Tibet mini-pigs with black hair can be used as the animal model of clinical laser hair removal.
Animals ; Hair Follicle ; radiation effects ; ultrastructure ; Hair Removal ; methods ; Lasers, Solid-State ; therapeutic use ; Microscopy, Electron, Transmission ; Swine ; Tibet
8.Expression of human estrogen receptor alpha and beta in Escherichia coli.
Lu LUO ; Shu-juan ZHAO ; Zheng-tao WANG ; Zhi-bi HU
Acta Pharmaceutica Sinica 2012;47(10):1399-1402
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
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Escherichia coli
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metabolism
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Estrogen Receptor alpha
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genetics
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metabolism
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Estrogen Receptor beta
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genetics
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metabolism
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Genetic Vectors
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Humans
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Protein Binding
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Recombinant Proteins
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genetics
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metabolism
9.Determination of calycosin-7-O-beta-D-glucopyranoside in radix astragali by HPLC.
Chun-xia LUO ; Ping-chuan LIN ; Li-hua GU ; Tao WU ; Da-zheng WU ; Zheng-tao WANG ; Zhi-bi HU
China Journal of Chinese Materia Medica 2003;28(7):603-606
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