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ObjectiveTo investigate the mechanism of Atractylodis Macrocephalae Rhizoma（AMR） in the treatment of slow-transmission constipation（STC） by observing the effects of AMR on short-chain fatty acids and intestinal barries in STC mice. MethodForty-eight male KM mice were randomly divided into blank group， model group， AMR low-， medium-， high-dose groups（2.5， 5， 10 g·kg-1） and mosapride group（2.5 mg·kg-1）. Except for the blank group， all groups were gavaged with loperamide suspension（5 mg·kg-1） twice daily for 14 d to construct the STC mouse model. At the same time， each drug administration group was given the corresponding drug by gavage for consecutive 14 d， the blank and model groups were gavaged with equal volume of distilled water. The effects of the treatment of AMR on body mass， defecation frequency， fecal water content and intestinal propulsion rate of mice were observed， the pathological changes of mouse colon were observed by hematoxylin-eosin（HE） staining and periodic acid-Schiff（PAS） staining， the levels of gastrin（GAS） and motilin（MTL） in serum were detected by enzyme-linked immunosorbent assay（ELISA）， gas chromatography-mass spectrometry（GC-MS） was used to detect the contents of short-chain fatty acids（SCFAs） in mouse feces， real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） and Western blot were used to determine the mRNA and protein expression levels of zonula occludens-1（ZO-1）， Occludin， and Claudin-1 in the colon of mice. ResultCompared with the blank group， the body mass， defecation frequency， fecal water content and intestinal propulsion rate of mice in the model group were significantly decreased（P<0.05， P<0.01）， the arrangement of colonic tissues was disordered， and the number of goblet cells was reduced， the levels of GAS and MTL in serum were significantly decreased（P<0.01）， and the levels of SCFAs in the feces were on a decreasing trend， with the contents of acetic acid， propionic acid， butyric acid， isobutyric acid and valeric acid were significantly decreased（P<0.05， P<0.01）， the mRNA and protein expression levels of ZO-1， Occludin and Claudin-1 in the colonic tissues were significantly decreased（P<0.01）. The above results suggested that STC mouse model was successfully constructed. Compared with the model group， the body mass， defecation frequency， fecal water content and intestinal propulsion rate of mice in AMP administration groups all increased significantly（P<0.05， P<0.01）， the mucosal layer of the colonic tissues was structurally intact without obvious damage， and the number of goblet cells increased， serum levels of GAS and MTL were significantly increased（P<0.01）， the contents of SCFAs in the feces were all on a rising trend， with the contents of acetic， propionic， butyric and isobutyric acids rising significantly（P<0.05， P<0.01）， the mRNA and protein expression levels of ZO-1， Occludin and Claudin-1 in the colonic tissues were significantly increased（P<0.05， P<0.01）. ConclusionAMR is able to improve the constipation symptoms in STC mice， and its mechanism may be related to increasing the contents of SCFAs in the intestine as well as promoting the mRNA and protein expression levels of ZO-1， Occludin and Claudin-1 in the colon.
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OBJECTIVE:To establish the quality standard of Hirudo nipponica freeze-dried powder(called"freeze-dried powder"for short),and to provide reference for controlling its quality. METHODS:A total of 3 batches of freeze-dried powder were collected,identified and tested according to the requirements of H. nipponica stated in 2015 edition of Chinese Pharmacopoeia(part Ⅰ)(shorted for pharmacopoeia);the antithrombin activity was also analyzed. The maximum tolerated dose (MTD)was used to investigate the toxicity. The stability was determined by designing temperature,humidity and strong light exposure tests. RESULTS:In the TLC of test sample,the same red spots were found in the corresponding location of the control drug chromatogram,and the same orange-red fluorescence spots were shown under the UV light(365 nm). Average content of moisture in 3 batches of samples was 2.61%,and the levels of total ash,acid-insoluble ash,pH aflatoxin and antithrombin activity were 2.83%,0.38%,6.92,0.28 μg/kg and 257.0 U/g,respectively. The content of Pb,Cd,As and Cu were in line with the requirements of pharmacopoeia except that the content of Hg was slightly higher than lower limit of H. nipponica in pharmacopoeia. Results of MTD showed that no death and ADR was found in mice after giving 26.4 g/kg freeze-dried powder by the amount of crude drug,which was 58 times as large as the maximum dosage that the pharmacopoeia described. Under the condition of 20, 40 ℃ and strong light exposure [(4 500±500)Lx],the anticoagulase activity of freeze-dried powder decreased significantly over time,while the anticoagulase activity of freeze-dried powder stored at 40 ℃ for 6 months was in line with the requirements of pharmacopoeia. Under the condition of high humidity(relative humidity were 90%,75%),freeze-dried powder showed a strong hygroscopicity. CONCLUSIONS:Established quality evaluation standard for freeze-dried powder according to pharmacopoeia standard could be used to control its quality.
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Objective:To quantitatively analyze the water content in Pinellia ternata by near-infrared spectroscopy to plant seed-ling cultivation and determine the quality of Pinellia ternata.Methods: Near-infrared spectroscopy was used to establish the model , and the moisture was measured by the drying method .The quantitative model was established by partial least squares method .Results:The quantitative model was established with the correlation coefficient of 0.9903 and the RMSECV of 0.205.The internal cross valida-tion and the external verification were both less than 1.0％.Conclusion:The established model can accurately predict the content of water in Pinellia ternata, which can be used for planting seedling cultivation , medicine storage management and quality control .
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This study was aimed to identify Bletilla Striata (Thunb.) Reichb.f.and Bletilla Formosana (Hayata) Schltr.by ITS2 sequence.The leaves of 38 samples of Bletilla striata and Bletillaformosana from Yunnan,Hubei,Guizhou,Hunan and Sichuan province were used as experiment materials.The total DNA was extracted.Internal transcribed spacer 2 (ITS2) sequences were obtained by PCR.All of the ITS2 sequences were checked.The 8 ITS2 sequences from two species were downloaded from GenBank.The intraspecific and interspecific Kimura-2-parameter (K2P) distances of Bletilla striata and Bletilla formosana were calculated by MEGAS.0.And neighbor-joining (NJ) tree was constructed.The results showed that the full-length sequences of ITS2 from Bletilla striata and Bletillaformosana were 259 bp,with a total of 14 variable sites.The maximum intraspecific K2P distance of Bletilla striata and Bletillaformosana was 0.008,while the minimum interspecific K2P distance was 0.040.The ITS2 secondary structure showed that different origins of Bletilla striata were gathered together and could be distinguished obviously from Bletilla formosana by NJ tree.It was concluded that ITS2 sequence was able to identify Bletilla striata and Bletillaformosana quickly and accurately.
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Objective: To quantitatively analyze the content of glucose in glucose injection by near infrared spectroscopy to control the quality of the product.Methods: A quantitative model was established by near infrared reflectance spectroscopy and the injection came from pharmaceutical enterprises with different concentrations of glucose and the solution samples came from laboratories with different concentrations of glucose.The liquid sample accessories were selected, a quantitative model was established by a partial least squares method, and the effect of environmental temperature on the model was studied as well.Results: According to the quantitative model, the correlation coefficient reached up to 0.999 3, and RMS deviation (RMSECV) was 0.077 4.In the verification test, the results of various liquid formulas containing glucose were similar to those of the laboratory results.The prediction error was less than 5% within the temperature range of 20-35℃.Conclusion: The model of near-infrared partial least squares (PLS) can accurately predict the content of glucose in glucose products without sodium chloride, and can be used for the quality control of glucose intermediates and the finished products in the large infusion manufacturers
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Objective To explore the application value of DOSE index score in the peitients with chronic obstructive pulmonary disease.Methods 122 cases of plateau COPD patients were followed up for 12 months,and we recorded and analyzed the patient's health and life.We also recorde FEV 1 and DOSE scores of the patients with COPD,and record the COPD risk events,including the number of respiratory failure and death,and the times of hospitalization,total such confinement,outpatient expenses,hospitalization expenses,mMRC,and scored in the number of exacerbations,etc.Results The DOSE index score was negatively correlated with FEV1％ pred (r=0.73,P ＜ 0.05) for 122 COPD patients,and were positively correlated with mMRC (r=085,P ＜ 0.01),the annual number of exacerbations (r=0.71,P ＜ 0.01),respiratory failure (r=0.65,P ＜ 0.01),heart failure (r=0.50,P ＜ 0.01),number of outpatient service (r=0.12,P＜0.01),hospitalization time (r=0.70,P＜0.01),the totalsuchconfinement (r=0.66,P＜0.01),outpatient expenses (r=0.13,P＞ 0.13),hospitalization expenses (r=0.65,P＜0.01).ROC curve was used to analyzed the cut-off point and curve area of COPD DOSE index.Conclusion The DOSE index is a simple COPD assessment tools,and is closely related to the prognosis of patients and health.
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Objective:To assess the genetic diversity and phylogenetic relationship of Desmodium styraeifolium from fifteen regions of Guangdong and Guangxi Zhuang autonomous region. Methods:The molecular technique ISSR((inter-simple sequence repeat))was applied to investigate the genetic diversity of Desmodium styraeifolium from fifteen regions of Guangdong and Guangxi Zhuang autonomous region. The data was analyzed with Popgene 1. 32,and a cluster diagram was presented by UPGMA. Results:Totally 51 amplified fragments were obtained by 7 ISSR primers. The results analyzed by Popgene 1. 32 showed that the Shannon diversity index(I)was 0. 3,the NEI’s genetic diversity coefficient(H)was 0. 246 4,the coefficient of genetic differentiation (GST)was 0. 123 8,and the gene flow(Nm)was 3. 539 7. Conclusion:The above mentioned results exhibit that Desmodium styraeifolium from Guangdong,Guangxi and some wild herbal populations has high genetic diversity. The clustering results illustrate that the genetic distance of Desmodium styraeifolium originated from Guangdong and Guangxi is related with geographic distance.
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Compared with that for botanical drugs and animal-derived drugs, the identification study for mineral traditional Chi-nese medicines is relatively weak. The traditional identification methods can’ t meet the quality control requirements of mineral tradi-tional Chinese medicines, and the application of modern analysis techniques are needed urgently in the systematic research of mineral traditional Chinese medicines. In the paper, the identification of traditional methods combined with some modern analysis techniques such as X-ray diffraction, near infrared spectroscopy and Raman spectroscopy for mineral traditional Chinese medicines was summarized and analyzed to provide basic idea and methods for the systematic identification construction of mineral traditional Chinese medicines.
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Objective:To explore a new method to identify Sparganium stoloniferum and its adulterants by ITS2 regions. Methods:Eight samples of Sparganium stoloniferum and its adulterants were collected with five species, and 6 species with 23 ITS2 sequence of Sparganium stoloniferum and its adulterants were downloaded from Genbank. The intraspecific and interspecific K2P distances of Spar-ganium stoloniferum and its adulterants were calculated by MEGA5. 0, and the phylogenetic tree was constructed by MEGA 5. 0. Re-sults:The maximum intraspecific K2P distance of Sparganium stoloniferum was 0. 038,while the minimum interspecific K2P distance was 0. 697. The phylogenetic tree showed that Plantago asiatica was different obviously from its adulterants. The different samples of Sparganium stoloniferum were gathered together and could be distinguished from its adulterants by the NJ tree. Conclusion: ITS2 se-quence is able to identify Sparganium stoloniferum and its adulterants correctly, which provides a new method for the identification of Sparganium stoloniferum.
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This study was aimed to establish X-ray diffraction (XRD) Fourier fingerprint of mineral Chinese medicine Actinolitum, in order to provide a new method for evaluating the quality of Actinolitum. Actinolitum samples were analyzed by the technology of powder XRD. And the XRD Fourier fingerprint was determined. Accord to the fingerprints and the intensity of each characteristic peak in XRD patterns of Actinolitum, the similarity of different samples were calculated using the law of cosines and the correlation coefficient method. Systematic cluster analysis was also used for the data. The results showed that XRD patterns of 10 certified products, 3 doping products and 2 counterfeits of Actinolitum were obtained. The geometric and topological characteristics of 10 certified products were consistent. XRD fingerprint of Actinolitum from 10 certified products had 18 common characteristic peaks. The similarity analysis showed that the similarity of XRD patterns common peak of certified products were higher among 15 samples (> 0.98). The similarity of doping products was slightly lower (0.85-0.97). And the counterfeits had the lowest similarity (< 0.2). These three had significant differences which can be distinguished. The results of cluster analysis were consistent with the similarity analysis results. It was concluded that XRD fingerprint had good specificity and feasible. It was accurate and reliable, which can be used to distinguish and evaluate Actinolitum.
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The studies on the chemical compositions and pharmacological actions of Chaenomeles speciosa Nakai were systemized and compared with those of the other plants of Chaenomeles in this paper. The pharmacological effects of the fruit of Chaenomeles inclu-ding anti-tumor, anti-oxidation, anti-inflammation and analgesia, antibacterial, hypoglycemic and hypolipidemic action and so on were reviewed to provide scientific basis for the further studies and utilization of Chaenomeles.
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Objective:To provide reference information for the further study and rational clinical application of Shuxuetong Injec-tion. Methods:Referred to the literatures on Shuxuetong Injection, the pharmacological actions and clinical application were intro-duced. Results:The development status of Shuxuetong Injection was summarized, and the pharmacological actions and clinical applica-tion were reviewed in the paper. Conclusion:When used alone or combined used with other medicines, Shuxuetong Injection have sig-nificant effects in the treatment of some cardiovascular and cerebrovascular diseases, especially ischemic stroke. However, the active constituents in Shuxuetong Injection still need further studies.
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Objective:To study the percutaneous permeability of quercetin in Ginkgo Biloba extract in vitro. Methods: Ginkgo Biloba extract was dissolved in 50% ethanol. The percutaneous permeability experiment through mouse skin was conducted using a modified Franz diffusion cell. The penetration amount of quercetin was determined by HPLC. Results:The percutaneous permeability of quercetin in the solution was approximately characterized by zero-order kinetics. Conclusion:Quercetin in Ginkgo Biloba extract can be absorbed through skin.
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Objective:To pharmacognostically identify Selaginellae uncinatae Herba to provide reference for the identification and utilization of Selaginellae uncinatae Herba. Methods:On the basis of observing the characters, structure and microscopic characteris-tics of the powder, Selaginella uncinata Herba was identified by a TLC method using amentoflavone as the reference substance. The contents of total ash, acid insoluble ash and alcohol-soluble extracts in Selaginellae uncinatae Herba were detected as well. Results:The morphological identification, microscopic identification and TLC identification of the herb was respectively established. The content limits of the total ash, acid insoluble ash and alcohol soluble extract were preliminarily determined. Conclusion:The studies provide reference for the identification and quality evaluation of Selaginellae uncinatae Herba.
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To establish seven kinds of minerals containing sulfate kind of near infrared spectral identification method of traditional Chinese medicine (TCM). 7 species of mineral medicine containing sulfate after crushing sieving, measure all the samples in 12 000-4 000 cm-1 section within the scope of the near infrared spectrum, spectrum signal by different pretreatment methods, after the screening of the different characteristics of the spectrum to extract the effective information, using cluster analysis method for qualitative identification. In 8 600-8 100 cm-1, 5 843-4 245 cm-1, 7 096-6 337 cm -1 section within the scope of the atlas signal after the vector normalization and multiple scattering correction, K-average clustering analysis to 20 batches sulfate kind of mineral medicine is divided into seven categories, the results of the analysis method and the traditional traits identification results are basically identical. This method is simple, fast, and can be used for these minerals containing sulfate class the qualitative identification and quality control of Chinese traditional medicine.
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Objective:To establish the ISSR fingerprint of Chrysanthemum morifolium cultivated in Futianhe area of Macheng county to guide the breeding of C. morifolium. Methods:Using the technology of ISSR molecular markers and the software of SPSS 15. 0, the coefficient matrix of Jaccard was established and the tree graph of the genetic relationship of the breeds of C. morifolium cultivated in Fu-tianhe area was built to analyze the respective genetic relationship and features. Results:By ISSR analysis, it confirmed that C. morifoli-um in Futianhe area had long genetic distance with the other white chrysanthemum breeds, and it could be considered as an individual breed. Conclusion:The ISSR map can be used to identify the breed of C. morifolium cultivated in Futianhe area.
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Objective:To develop a qualitative analysis model for the fast identification of Dens Draconis and its adulterants by NIR correlation coefficient method. Methods:On the basis of the traditional morphological identification, the spectra were collected u-sing the fiber accessory of a near-infrared spectroradiometer. The reference spectra were set up using the NIR spectra of certified Dens Draconis. The characteristic spectral section was chosen and the appropriate threshold was set to establish a qualitative analysis model for the rapid identification of Dens Draconis and its adulterant. Results:The spectral section of 5 000-4 200 cm-1 was selected as the characteristic spectral section, the correlation coefficient of Dens Draconis and its adulterant was calculated in training set samples, and 92. 67% was used as the threshold. Totally 10 batches of validation set samples were validated the qualitative analysis model, and the prediction accuracy was 90%. Conclusion:The method has good prediction ability, and can be used in the rapid identification of Dens Draconis and its adulterant.
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Objective: To analyze the influencing factors in thrombin titration for the determination of anticoagulant activity of Whitmania Pigra Whitman. Methods: The white porcelain plates were used as the titration carriers instead of tubes in the titration ( called white porcelain method for short) . The effect of different carriers, interval time of titration and thrombin concentration on the results of anticoagulant activity test was studied. Results:Under the same conditions, the anticoagulant activity was more accurate and stable using white porcelain method. Using white porcelain method with 20 u·ml-1 or 10 u·ml-1 as the thrombin concentration and titrating 5μl each time, once every minute, the thrombin consumption volume was linear with the sample concentration within the range of 0. 125-0. 333 g·ml-1(r20 =0. 961 and r10 =0. 992), and the anticoagulant activity respectively was (33. 08 ± 2. 64) and (31. 24 ±1.32) u·g-1(RSD20 =8.0% and RSD10 =4.2%). As for a certain sample concentration (0.333 g·ml-1), the theoretical error of determination was not more than 10% and 5%. Conclusion:The improved white porcelain method is more suitable for determining anticoagulant activity of Whitmania Pigra Whitman with more stable results and accurate end point states than tube method. Under the conditions of 10 u·ml-1 thrombin concentration, titrating 5μl each time, once every minute, the linearity, accuracy and precision are all promising.
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Objective:To seek the best part and optimal culture medium for the tissue culture of Bletilla striata. Methods: The effects of hormone concentration and proportion on the induced differentiation, propagation and rootage of Bletilla striata were investiga-ted. Results:The radicle of Bletilla striata was the best part to induce the clustered shoots with the optimal culture medium of 1/2MS+1. 0 mg·L-1 6-BA+2. 0 mg·L-1 NAA. The best hormone concentration for inducing the clustered shoots was MS+1. 0 mg·-1 L 6-BA+0. 05 mg·L-1 NAA, and the optimal rooting medium was 1/2MS+0. 5 mg·L-1 NAA. Conclusion: The tissue culture system for Bletilla striata is established.
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Theancientandmodernprocessingmethods,activeingredientresearch,andqualitycomparisonbeforeandafterthe processing of medicinal leech were reviewed in the paper. Combined with the results of anticoagulation test on Whitmania pigra Whit-man, the article drew a conclusion that different varieties of medicinal leech had different active ingredients of anticoagulation, Whitma-nia pigra Whitman had active ingredients of anticoagulation with promising heat stability, and the processing technology should be dealt with according to the clinic use.