Effects and mechanisms of Tujia medicine musk needle therapy on improving cognitive dysfunction in ischemic stroke model rats / 中国药房

OBJECTIVE To study the improvement effects and mechanisms of Tujia medicine musk needle therapy on cognitive dysfunction in ischemic stroke model rats. METHODS Totally 44 rats were randomly divided into sham operation group， model group， musk needle treatment group and ordinary acupuncture group， with 11 rats in each group. Except for the sham operation group， ischemic stroke model was induced by modified suture method in other groups. After modeling， musk needle treatment group and the ordinary acupuncture group were treated with Tujia musk needle （containing 3 mg of artificial musk） and traditional filiform needle respectively to intervene in the muscle layer of the contralateral scalp motor area， with an intervention duration of 3 courses. The sham operation group and model group were not given any treatment. The neurological deficits score in rats were recorded and Morris water maze behavioral tests were conducted. The morphology of neurons in the cortical area of rats was observed， and the expression of DCX/BrdU and NeuN/BrdU co-labeled cells in the ischemic subependymal area was observed. The plasma levels of hypoxia-inducible factor-1α （HIF-1α） and vascular endothelial growth factor（VEGF） in rats were tested. RESULTS Compared with sham operation group， neurological deficit score of model group was increased significantly， escape latency prolonged significantly， and the times of crossing platform significantly reduced （P＜0.05）； the neuronal structure was significantly damaged， and the number of surrounding Nissl bodies decreased； the number of DCX/BrdU and NeuN/BrdU co- labeled cells in the ischemic subependymal area were significantly increased （P＜0.05）； the levels of HIF-1α and VEGF in plasma were significantly increased （P＜0.05）. Compared with model group， neurological deficits score， escape latency， the times of crossing platform were all reversed significantly in musk needle treatment group and ordinary acupuncture group （P＜0.05）； the neuronal structure was improved， and the number of Nissl bodies increased； the number of DCX/BrdU and NeuN/BrdU co-labeled cells in the ischemic subependymal area were significantly increased （P＜0.05）； the plasma levels of HIF-1α and VEGF were significantly increased （P＜0.05）. Compared with ordinary acupuncture group， the plasma level of HIF-1α was reduced （the difference was not statistically significant）， while the level of VEGF was significantly increased （P＜0.05）. CONCLUSIONS Tujia medicine musk needle therapy can significantly improve the cognitive dysfunction in ischemic stroke model rats， and its mechanism of action may be associated with promoting migration and differentiation of neural stem cell in ischemic subependymal area， preventing the excessive release of HIF-1α and increasing the expression of VEGF.

Metabolomics study of Berberidis Radix in intervening ulcerative colitis based on UPLC-Q-TOF-MS / 中国中药杂志

The effect of Tujia medicine Berberidis Radix on endogenous metabolites in the serum and feces of mice with ulcerative colitis(UC) induced by dextran sulfate sodium(DSS) was analyzed by metabolomics technology to explore the metabolic pathway and underlying mechanism of Berberidis Radix in the intervention of UC. The UC model was induced in mice by DSS. Body weight, disease activity index(DAI), and colon length were recorded. The levels of tumor necrosis factor-α(TNF-α) and interleukin-10(IL-10) in colon tissues were determined by ELISA. The levels of endogenous metabolites in the serum and feces were detected by ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS). Principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were employed to characterize and screen differential metabolites. The potential metabolic pathways were analyzed by MetaboAnalyst 5.0. The results showed that Berberidis Radix could significantly improve the symptoms of UC mice and increase the level of the anti-inflammatory factor IL-10. A total of 56 and 43 differential metabolites were identified in the serum and feces, respectively, belonging to lipids, amino acids, fatty acids, etc. After the intervention by Berberidis Radix, the metabolic disorder gradually recovered. The involved metabolic pathways included biosynthesis of phenylalanine, tyrosine, and tryptophan, linoleic acid metabolism, phenylalanine metabolism, and glycerophospholipid metabolism. Berberidis Radix can alleviate the symptoms of mice with DSS-induced UC, and the mechanism may be closely related to the re-gulation of lipid metabolism, amino acid metabolism, and energy metabolism.

Content determination of Chikusetsu saponin Ⅳa of root, stem and leaf in Panacis japonici rhizoma / 国际中医中药杂志

Objective:To determine the content of Chikusetsu saponin Ⅳa in the roots, stems and leaves in Panacis japonici rhizoma, and explore its development value. Methods:By using High Performance Lliquid Chromatography (HPLC) method to determine the content of Chikusetsu saponin Ⅳa in the roots, stems and leave of Panacis japonici rhizoma. Inertsil ODS-sp (4.6 mm×150 mm, 5 μm) was used as the column and the acetonitrile-0.2% phosphoric acid (35:65) as the mobile phase. The flow rate was 1 ml/min with the temperature 35 ℃ under wavelength 210 nm. Results:The linear ranged from 1-10 μg of Chikusetsu saponin Ⅳa was good ( r=0.999 5). The content of Chikusetsu saponin Ⅳa in the roof, stem and leaf were 2.169%, 0.269%, 0.169%, respectively. Conclusions:The roof of Panacis japonici rhizoma contains the highest level of Chikusetsu saponin Ⅳa followed by stem and leaf. The roof part could be used as the medical part and its stem and leaf are also valuable for further development.

Study on Chemical Constituents of Petroleum Ether Part from Whole Herbs of Pteris vittata / 中国药房

OBJECTIVE： To study the chemical constituents from the petroleum ether part from whole herbs of Pteris vittata. METHODS： The petroleum ether part from whole herbs of P. vittata was separated and isolated by silica gel column， gel column， recrystallization and TLC. The structures of the compounds were identified according to physicochemical properties and spectrum data （1H-NMR and 13C-NMR）. RESULTS： A total of 11 compounds were isolated and identified from the petroleum ether part from whole herbs of P. vittata， as （2R）-acetyl pterosin B （Ⅰ）， palmitic acid （Ⅱ）， hop-22（29）-ene （Ⅲ）， epifriedelanol （Ⅳ）， lupenone （Ⅴ）， olean-18-en-3-one （Ⅵ）， stigmasterol （Ⅶ）， β-sitosterol （Ⅷ）， 22-hydroxyhopane （Ⅸ）， ergosterol （Ⅹ）， β-sitosterol acetate （Ⅺ）. CONCLUSIONS： Compounds Ⅰ-Ⅶ and Ⅸ-Ⅺ are isolated from this plant for the first time， and can provide theoretic reference for further studying bioactive pharmacodynamic substances in P. vittata and enriching chemical component data.

Therapeutic effects of Bianyanning decoction on acute pharyngitis in rats and its mechanism / 中国应用生理学杂志

OBJECTIVE@#To investigate the therapeutic effects and mechanisms of Bianyanning on acute pharyngitis in rats, and to provide evidence and experimental data for its clinical application.@*METHODS@#The acute pharyngitis of rats was induced by spraying ammonia directly to their throat. The model rats were randomly divided into model control group, the high-, medium- and low-dose group of Bianyanning, while normal rats were used as control group, 10 in each group. After the corresponding drug treatment, the symptoms and manifestations of each group were observed and recorded; 24 hours after last gavaging, blood samples of each group were collected from the abdominal aorta. The serum contents of interleukin 1-beta (IL-1β) and tumor necrosis factor alpha (TNF-α) were detected by ELISA. HE method was used to observe the characteristic of the lung tissues and the transmission electron microscopy method was used to observe the trachea cilia.@*RESULTS@#After the treatment, compared with the model control group, the high-, medium- and low-dose group of Bianyanning, the symptoms of acute pharyngitis such as inflamed and congestive throat were relieved obviously. The morphological changes of lung and bronchus tissues were apparently improved. The contents of IL-1β and TNF-α in serum were decreased significantly.@*CONCLUSION@#Compound Bianyanning can promote the recovering process of acute pharyngitis, improve the morphology of lungs and bronchus, which may be related to inhibiting the releasing of the IL-1β and TNF-α in serum.

Discussion on Difference of Different Compatibility Ratios of Astragali Radix and Puerariae Lobatae Radix Based on Content Analysis / 中国中医药信息杂志

Objective To study the contents of puerarin in decoction with different compatibility ratios of Astragali Radix and Puerariae Lobatae Radix; To verify the rationality of compatibility ratios of Huangqi Gegen Decoction and Yu y e Decoction; To provide references for clinical medication. Methods Different compatibility ratios of Astragali Radix and Puerariae Lobatae Radix were set as 0:1, 1:1, 2:1, 3:1, 4:1, 1:2, 1:3, and 1:4. SinoChrom ODS-BP (4.6 mm × 250 mm, 5 μm) was used as the chromatographic column to detect the contents of puerarin;methanol-0.1% phosphoric acid (35:65) was used as the mobile phase with 0.5 mL/min flow rate; sample volume was 20 μL; the wavelength was 250 nm. Results The regression equation of puerarin was Y=66 449.269 1X-175 665.663 1 (r=0.999 6) in the range of 5 to 300 μg/mL, showing a good linear relationship. The repeatability, stability and recovery rate were fine as well. The contents of puerarin were (2.506 7±0.025 8)%, (2.526 7±0.071 2)%, (2.863 3± 0.086 4)%, (2.956 7±0.119 6)%, (2.835 0±0.078 7)%, (2.480 0±0.072 4)%, (2.530 0±0.064 8)%, and (2.183 3±0.128 9)% in different compatibility ratios of Astragali Radix and Puerariae Lobatae Radix with 0:1, 1:1, 2:1, 3:1, 4:1, 1:2, 1:3, and 1:4, respectively. Conclusion When the compatibility ratios of Astragali Radix and Puerariae Lobatae Radix are 2:1. 3:1, and 4:1, the contents of puerarin in decoction are the highest. This study verifies that the compatibility ratios of Astragali Radix and Puerariae Lobatae Radix in classical prescriptions are rational, which can be the optimal compatibility ratios in clinic.

Percutaneous Absorption in Vitro of Jiegugao Blended and Pasted by Commonly Used ;Ointment Matrices in Tujia Minority / 中国中医药信息杂志

Objective To establish a method to simultaneously determine syringing and isofraxidin by HPLC;To investigate the features of percutaneous absorption in vitro of Jiegugao blended and pasted by white vinegar, honey and vaseline; To discuss the mechanism of commonly used ointment matrices in Tujia Minority. Methods Rat abdomen skin in vitro was as transdermal barrier;the modified Franz diffusion pool was used to simulate human skin medication; the content of syringin and isoprofen was determined by HPLC; the percutaneous absorption equation was established and the related parameters, such as cumulative permeation rate and permeation rate, were calculated. Results When using Syncronis C18 (250 mm × 4.6 mm, 5 μm) as chromatographic column, acetonitrile-0.1%phosphoric acid as mobile phase, 1.0 mL/min as perfusion speed and 265 nm as determine wavelength, regression equation of syringingwas A=10 686.454 6C+1565.778 8 (r=1.000 0), regression equation of isofraxidin was A=12 297.305 4C-5913.729 9 (r=0.999 9). Cumulative permeation quantity of syringing in Jiegugao blended and pasted by white vinegar, honey, vaseline and blank were 7.549 2, 4.580 3, 3.890 8 and 5.378 4 μg?cm-2?h-1 respectively and permeation rate were 25.66%, 16.11%, 13.73% and 18.78%. Meanwhile, cumulative permeation quantity of isofraxidin were 2.536 9, 1.941 8, 1.178 2 and 2.293 6 μg?cm-2?h-1 respectively and permeation rate were 47.04%, 35.06%, 22.11%and 41.11%. Conclusion Using white vinegar as the ointment matrix can promote the percutaneous absorption of effective composition in Jiegugao blended. However, it will retard the percutaneous absorption of effective composition in Jiegugao when using honey and vaseline as the ointment matrices, but honey and vaseline can be used as a slow-release matrix.

Study on ecological environment and accompanying plants' community characteristics study of wild Panax japonicus in Enshi / 中国中药杂志

The paper is aimed to study the distribution, population density, soil conditions and community characteristics of accompanying plants' in Enshi sub-regional different areas, with a typical habitats investigation method. The results showed that the wild Panax japonicus mainly distributed in moist places under the forests, by streams, or secondary forests of high grass, within east longitude 29°-30°, north latitude 108°-110°and about 1 000-15 00 meters above sea level. The soils were mainly tide soil and humus with yellow-brown soil, yellow soil and red soil, and the humus thickness was5-30 centimeter, pH 6.0-6.8, the moisture content of 16.8%-24.2%, soil bulk density of 1.39-2.12. Its geographical vegetation types were mainly evergreen coniferous forest, evergreen-deciduous mixture broad leaved forest and evergreen coniferous forest mixed deciduous broad-leaved forest, including three levels community structure of arbors, shrubs and herbaceous; Its accompanying plants reached 86 families, 118 genera, 134 species of seed plants, the arbors included 15 families, 21 genera, 26 species and the dominant species community mainly Pinaceae such as Pinus massoniana, P. tabuliformis, P. henryi and Taxodiaceae such as Cunninghamia lanceolata, Cryptomeria fortunei etc. The shrubs included 39 families, 54 genera, 62 species with the dominant species such as Camellia oleifera, Kalopanax septemlobus, Akebia trifoliata, Trachycarpusfortunei, Rhamnus globosa, Smilax corbularia and so on. The herbaceous included 32 families, 43 genera, 46 species, and Ferns such as the black-footed Dryopteris, Dryopteris crassirhizom, Coniogramme affinis, Polystichum tripteron, Adiantum pedatum, Lunathyrium acrostichoides, Woodsia ilvensis and Woodwardia japonica were dominant species. The cover layer covered a large number of lichens and mosses. The wild P. japonicus can be found among the P. massoniana, P. tabuliformis, P. henryi, lichens and mosses. These may indicate that the wild P. japonicusin Enshi requires higher demands on the ecological environment, its accompanying plants are mainly the tree layer-shrub layer-herb layer, and vertical structure is obvious. The study provides a basis for domestication and conservation of P. japonicus resources.

Comparison Analysis of Drug Instructions between Hospital Directory and OTC Standard Model Instruc-tions in 2014 / 中国药房

OBJECTIVE:To analyze the differences of drug instructions between hospital directory and OTC standard model in-structions,and to provide reference for enhancing instruction management and reducing the safety risk of clinical drug use. METH-ODS:1 324 drugs of hospital directory in a hospital in 2014 were compared with OTC directory from CFDA websites. The instruc-tion of drug types included in OTC directory were compared OTC model instruction. According to the degree of risks which the dif-ferences may bring,differences were divided into four levels for analysis as negligible,general,important and severe. RESULTS:244 drugs belonged to OTC,of which 32.38%were different from standard model instructions. The four risk levels rates of negligi-ble,general,important and severe accounted for 29.11%,34.18%,7.59% and 29.11%,respectively. Among important risk,the difference of“indication limit”occupied the highest proportion,being 50.00%. Among severe risk,the difference of“forbidden for special disease”and“forbidden for pregnant women”accounted for 43.48% and 39.13%. CONCLUSIONS:There are problems, such as the absence of important medication information,statement conflicts. The hospital and administration departments should en-hance the standard management of drug instruction to guarantee safe and rational drug use in the clinic.

Clinical Analysis of 82 Cases of Disulfiram-like Reaction in Our Hospital / 中国药房

OBJECTIVE:To provide reference for rational drug use in the clinic. METHODS:The cases of disulfiram-like reac-tion collected from the First Affiliated Hospital of Guangzhou University of TCM from July 1st in 2012 to June 30st in 2014,were analyzed and compared retrospectively. The epidemiological characteristics,causes,clinical characteristics and treatment methods were summarized. RESULTS:Disulfiram-like reaction mainly occurred in male(86.59%),mainly in young and middle-aged men aged 21-50 years old (78.05%). It might be induced by using cephalosporin antibiotics (as cefoperazone,cefperazone/sulbactam, etc.),nitroimidazole (as metronidazole,tinidazole,etc.),drinking or contacting beverage,food or drug containing ethanol. This reaction involved the skin and its accessories system,circulatory system,digestive system,nervous system,respiratory system, etc. Its clinical manifestations mainly were skin flush,chest tightness,shortness of breath,palpitation,nausea,vomiting,dizzi-ness,headache,etc.,occasional confusion,drowsiness,xerostomia,etc. Response to the severe disulfiram-like reaction,the foun-dation treatment combined with symptomatic treatment showed a good effect with remission of symptoms in 1 to 3 hours and no deaths. CONCLUSIONS:When the drugs with mercapto-1-methyltetrazole perssad as cephalosporin antibiotics,nitro imidazoles, etc.,are used,we should avoid drinking or taking drugs or food containing ethanol,to prevent the occurrence of disulfiram-like re-action.

Establishment and assessment of the diarrhea rat model of liver-QI stagnation with spleen deficiency / 中国实验动物学报

Objective To establish a diarrhea rat model using multiple-stimulating factors and choosing the best indexes to assess whether the model is consistent with the disease characteristics of liver -QI stagnation with spleen deficien-cy in traditional Chinese medicine .Methods Newborn SD rats were randomly divided into model group ( n=20 ) and control group (n=10).The rats of model group were stimulated by maternal separation , restraint stress and rectum acetic acid irritation, while the rats in control group were fed as normal .Weight changes, rectal sensitivity, Bristol scores and wa-ter content of feces and histology of the colon tissues were used as evaluation indexes to assess whether the model meets the demands for further studies .Results The rats in the model group showed loss of appetite , increase of water intake and u-rine reduction .Some rats showed increased activity , and even mania .Bristol scores and water content of feces were signifi-cantly higher than that of the control group , and the rectal sensitivity was significantly increased .The colon mucosa showed slightly thickened submucosal layer and mild inflammatory cell infiltration in the model rats .Conclusions The rat model established in this study is better to simulate the clinical manifestation of liver -QI stagnation with spleen deficiency in Chi-nese medicine , and may meet the demands of related researches of this disease .

Study on processing method of Cistanche tubulosa / 中国中药杂志

<p><b>OBJECTIVE</b>To establish the processing method of Cistanche tubulosa decoction pieces.</p><p><b>METHOD</b>The orthogonal test of four factors and three levels was used to optimize the main factors in the process of fresh C. tubulosa decoction pieces processing, including the thickness, temperature, and the time for inactivation of the enzyme in the plant. The result showed that the optimized condition was that fresh C. tubulosa was cut into 4 mm thickness, and heated at 70 degrees C for inactivation the enzyme in the plant for 6 min. Moreover, the optimized method was compared with the method of insolation and traditional dried method.</p><p><b>RESULT</b>The content of echinacoside in the C. tubulosa decoction piece by the optimized method was 7.3 times of that dried by insolation, and 12.8 times of that by traditional dry method; the content of verbascoside was 6. 5 and 14. 9 times of that dried by insolation and by traditional dry method, respectively; the content of galactitol was 7.1 and 13.2 times of that dried by insolation and by traditional dry method, respectively.</p><p><b>CONCLUSION</b>The quality of C. tubulosa decoction pieces could be improved by this method, and its crud drug could be saved, which would protect the source of the mild Herba Cistanche, and produced the better economic and ecological benefits.</p>

Cloning of Human canstatin gene and expression of its recombinant protein / 中国医学科学院学报

<p><b>OBJECTIVE</b>To clone human canstatin gene and express its recombinant protein.</p><p><b>METHODS</b>The total RNA was extracted from human placenta. The canstatin gene fragment was synthesized and amplified from the total RNA by RT-PCR. The resulting product was cloned into pUCm-T vector and transformed into E.coli DH5alpha through electroporation. The gene was sequenced by the Sanger Dideoxy-mediated chain-termination method, and then the canstatin cDNA was cloned into the BamHI and HindIII sites of plasmid pET-22b (+) and transformed into E.coli BL21 where it was induced to express proteins by isopropyl-1-thio-b-Dgalactopyranoside (IPTG).</p><p><b>RESULTS</b>The extracted total RNA was separated into three clear bands indicating 28S, 18S, and 5S after electrophoresis. The canstatin gene fragment was synthesized and amplified from the total RNA by RT-PCR. The resulting products were cloned into pUCm-T vectors, and then were transformed into E.coli DHSa. After an over night culture, both blue and white colonies were found on the agar plate. Six white colonies were selected and cut by BamHI and HindIII. The plasmids DNA in one white colony showed one band near the location of primary plasmid after digested by BamHI and two bands near the locations of primary plasmid and objective gene fragment after digested by HindIII. The cloned gene in this white colony was sequenced and demonstrated to have the same sequence as that of canstatin gene in GenBank. Then canstatin cDNA was cut down from pUCm-T with BamHI and HindIII and ligated into the vector pET-22b (+). The resultant plasmid pET-22b (+)/canstatin was then transformed into E.coli BL21. White colonies were found on LB agar plate. Seven of them were selected and their plasmids were digested with both BamHI and HindIII. After electrophoresis, all selected colonies showed two specific bands, one was found near the location of primary plasmids, and the other near that of objective gene fragment. After IPTG induction, there was a new protein band about Mr 24 000 on SDS-PAGE. As estimated by densitometry, the percentage of the expressed product over total bacterial proteins was 18.2%, 18.8%, 23.0% and 23.4%, respectively, 1, 2, 3, and 4 hours after induction.</p><p><b>CONCLUSION</b>Human canstatin gene was successfully cloned and its recombinant proteins were expressed in this study.</p>