Determination of the Contents of Three Lignans in Dendrobium fimbriatum Hook / 广州中医药大学学报

Objective To establish the method for content determination of three lignans of Dendrobium Fimbriatum Hook..Methods The lignans in Dendrobium tasselii were identified by high-performance liquid chromatography/multi-stage mass spectrometry(HPLC-ESI/MSn)coupled with ultraviolet absorption spectrometry(UV)coupled with retention time localization of high-performance liquid chromatography(HPLC).The separation was carried out on a Kromasil 100-5 C18 column(4.6 mm×250 mm,5 μm)using a gradient elution of acetonitrile-0.1%formic acid solution as the mobile phase,the volume flow rate was 0.8 mL·min-1 and the column temperature was 35℃,and the mass spectrometry was performed using an ESI ion source with the data collected in the negative ion mode.The HPLC content was determined on the same column as that of MS analysis,with the mobile phase methanol + acetonitrile(V/V=1∶1)-0.01 mol/L ammonium acetate solution,gradient elution,flow rate of 0.8 mL·min-1,column temperature of 40℃,and detection wavelength of 215 nm.Results Syringaresinol di-O-glucoside and(-)-Syringaresinol 4-O-β-D-glucopyranoside and DL-Syringaresinol were identified from Dendrobium fimbriatum Hook.,and the results of content determination showed that the linear ranges of above three components were respectively 0.1701-3.4020,0.1020-2.0400,0.0403-0.8060 μg(r≥0.9995),the average recoveries were in the range of 97.71%-101.67%,and the relative standard deviations(RSDs)were all less than 3.0%.The contents of Syringaresinol di-O-glucoside and(-)-Syringaresinol 4-O-β-D-glucopyranoside and DL-Syringaresinol in the 10 batches of samples were 0.7779-1.3852,0.0734-0.1966,0.0295-0.1882 mg·g-1.Conclusion This research method can provide a reference basis for the quality evaluation method of Dendrobium fimbriatum Hook..

Effect of heme oxygenase-1-mediated atorvastatin on macrophage polarization and cholesterol accumulation / 中国组织工程研究

BACKGROUND:Studies have shown that atorvastatin can up-regulate the expression of heme oxygenase-1 and enhance the anti-inflammation and anti-oxidative damage ability of cells.However,whether atorvastatin can regulate macrophage polarization,inhibit inflammation and reduce cholesterol accumulation by inducing heme oxygenase-1 expression remains unclear. OBJECTIVE:To investigate the effect of atorvastatin on polarization,inflammation and cholesterol content of oxidized low-density lipoprotein stimulated RAW264.7 macrophages by inducing heme oxygenase-1 expression and its related mechanism. METHODS:Firstly,RAW264.7 cells were randomly divided into six groups and incubated with different concentrations of atorvastatin for 24 hours.The expression of heme oxygenase-1 protein and cell activity were detected to explore the optimal dose of atorvastatin for subsequent studies.RAW264.7 cells were randomly divided into control group,atorvastatin group and heme oxygenase-1 inhibition group.Cells were preincubated with pure medium,atorvastatin 20 μmol/L and atorvastatin 20 μmol/L + zinc protoporphyrin IX 10 μmol/L for 24 hours,and then oxidized low-density lipoprotein 50 mg/L was added for 48 hours.The polarization of macrophages was detected by flow cytometry.The secretion of inflammatory factors such as transforming growth factor β,interleukin 10,interleukin 1β,and tumor necrosis factor α was detected by ELISA.The expression levels of heme oxygenase-1,LC3II,LC3I,P62,PPARγ and ABCA1 were detected by western blot assay.The intracellular cholesterol content was measured with the oxidose method and the accumulation degree of intracellular lipid droplets was evaluated by oil red O staining. RESULTS AND CONCLUSION:(1)Atorvastatin could induce the expression of heme oxygenase-1 protein in macrophages in a dose-dependent manner.(2)Oxidized low-density lipoprotein could induce macrophages to polarize towards M1,secrete proinflammatory factors,and increase the accumulation of intracellular cholesterol.(3)Compared with the control group,the heme oxygenase-1 protein expression of macrophages was increased after atorvastatin intervention,and the cells turned to M2-type polarization and mainly secreted anti-inflammatory factors such as transforming growth factor-β and interleukin-10.PPARγ,ABCA1,LC3II/I and other signal molecules reflecting cholesterol efflux and autophagy increased,and the contents of intracellular cholesterol and lipid droplets decreased significantly(P<0.05).(4)The heme oxygenase-1 inhibition group treated with zinc protoporphyrin IX significantly reversed the above changes in the atorvastatin group.(5)The results have shown that atorvastatin may promote the polarization of macrophages stimulated by oxidized low-density lipoprotein to M2 type and inhibit inflammation by up-regulating the expression of heme oxygenase-1 and by up-regulating PPARγ/ABCA1 signaling pathway and enhancing autophagy.Atorvastatin can increase the outflow of intracellular cholesterol and reduce the accumulation of intracellular lipids.

Qualitative and quantitative study of flavonoids in Notoginseng Radix et Rhizoma based on UPLC-Q-TOF-MS and HPLC-DAD / 中国中药杂志

The flavonoids in Panax notoginseng were qualitatively analyzed by ultra-high performance liquid chromatography-quadrupole-time of flight mass spectrometry(UPLC-Q-TOF-MS), and the content of three main flavonoids in P. notoginseng of different specifications and grades collected from different habitats was determined by HPLC-DAD. Flavonoids and anthocyanins were analyzed by UPLC-Q-TOF-MS/MS in the positive and negative ion modes, respectively. Twelve flavonoid glycosides and one anthocyanin glycoside in P. notoginseng were identified, but no flavonoid aglycones were detected. Among them, 12 compounds were identified in the underground part of P. notoginseng for the first time and eight compounds were first reported in this plant. Moreover, six and four compounds were identified in the Panax genus and the Araliaceae family for the first time, respectively. A method for simultaneous determination of three flavonoids in P. notoginseng was established by HPLC-DAD. The content of flavonoids in 721 P. notoginseng samples of 124 specifications and grades collected from 20 different habitats was simultaneously determined. Among three flavonoids determined, the content of quercetin-3-O-(2″-β-D-xylosyl)-β-D-galactoside was the highest with the average content in the tested samples of 161.0 μg·g~(-1). The content of compounds quercetin-3-O-hexosyl-hexoside and kaempferol-3-O-pentosyl-hexoside was relatively low, with the average content of 18.5 μg·g~(-1)(calculated as quercetin-3-O-sophoroside) and 49.4 μg·g~(-1)(calculated as kaempferol-3-O-sangbu diglycoside). There were significant differences in flavonoids content of samples from different production area. The content of flavonoids in spring P. notoginseng was significantly lower than that in winter P. notoginseng when the other influencing factors such as production areas, germplasm resources, and cultivation conditions were fixed. As for P. notoginseng of different specifications, the flavonoid content in the part connecting the taproot and the aboveground stem was significantly higher than that in other parts. The results of large-scale data showed that the flavonoid content gradually increased with the increase in the number of heads. There were significant differences between the flavonoid content in most specifications and grades, especially the 20-head P. notoginseng and countless head P. notoginseng, whose content was significantly lower and significantly higher than that of other specifications and grades, respectively. This study provides a scientific basis for the study of the effective components and quality control of P. notoginseng from the perspective of flavonoids.

Erratum: Author correction to 'Amino-functionalized poloxamer 407 with both mucoadhesive and thermosensitive properties: Preparation, characterization and application in vaginal drug delivery system' Acta Pharm Sin B 7 (2017) 593-602

[This corrects the article DOI: 10.1016/j.apsb.2017.03.002.].

Injectable hydrogel microspheres experimental research for the treatment of osteoarthritis / 中国修复重建外科杂志

OBJECTIVE@#To prepare a novel hyaluronic acid methacrylate (HAMA) hydrogel microspheres loaded polyhedral oligomeric silsesquioxane-diclofenac sodium (POSS-DS) patricles, then investigate its physicochemical characteristics and in vitro and in vivo biological properties.@*METHODS@#Using sulfhydryl POSS (POSS-SH) as a nano-construction platform, polyethylene glycol and DS were chemically linked through the "click chemistry" method to construct functional nanoparticle POSS-DS. The composition was analyzed by nuclear magnetic resonance spectroscopy and the morphology was characterized by transmission electron microscopy. In order to achieve drug sustained release, POSS-DS was encapsulated in HAMA, and hybrid hydrogel microspheres were prepared by microfluidic technology, namely HAMA@POSS-DS. The morphology of the hybrid hydrogel microspheres was characterized by optical microscope and scanning electron microscope. The in vitro degradation and drug release efficiency were observed. Cell counting kit 8 (CCK-8) and live/dead staining were used to detect the effect on chondrocyte proliferation. Moreover, a chondrocyte inflammation model was constructed and cultured with HAMA@POSS-DS. The relevant inflammatory indicators, including collagen type Ⅱ, aggrecan (AGG), matrix metalloproteinase 13 (MMP-13), recombinant A disintegrin and metalloproteinase with thrombospondin 5 (Adamts5), and recombinant tachykinin precursor 1 (TAC1) were detected by immunofluorescence staining and real-time fluorescence quantitative PCR, with normal cultured chondrocytes and the chondrocyte inflammation model without treatment as control group and blank group respectively to further evaluate their anti-inflammatory activity. Finally, by constructing a rat model of knee osteoarthritis, the effectiveness of HAMA@POSS-DS on osteoarthritis was evaluated by X-ray film and Micro-CT examination.@*RESULTS@#The overall particle size of POSS-DS nanoparticles was uniform with a diameter of about 100 nm. HAMA@POSS-DS hydrogel microspheres were opaque spheres with a diameter of about 100 μm and a spherical porous structure. The degradation period was 9 weeks, during which the loaded POSS-DS nanoparticles were slowly released. CCK-8 and live/dead staining showed no obvious cytotoxicity at HAMA@POSS-DS, and POSS-DS released by HAMA@POSS-DS significantly promoted cell proliferation (P<0.05). In the chondrocyte anti-inflammatory experiment, the relative expression of collagen type Ⅱ mRNA in HAMA@POSS-DS group was significantly higher than that in control group and blank group (P<0.05). The relative expression level of AGG mRNA was significantly higher than that of blank group (P<0.05). The relative expressions of MMP-13, Adamts5, and TAC1 mRNA in HAMA@POSS-DS group were significantly lower than those in blank group (P<0.05). In vivo experiments showed that the joint space width decreased after operation in rats with osteoarthritis, but HAMA@POSS-DS delayed the process of joint space narrowing and significantly improved the periarticular osteophytosis (P<0.05).@*CONCLUSION@#HAMA@POSS-DS can effectively regulate the local inflammatory microenvironment and significantly promote chondrocyte proliferation, which is conducive to promoting cartilage regeneration and repair in osteoarthritis.

Deoxycytidine kinase participates in the regulation of ionizing radiation induced ferroptosis in triple-negative breast cancer cells / 中华放射肿瘤学杂志

Objective:To investigate the regulatory effect of deoxycytidine kinase (dCK) on ionizing radiation (IR) induced ferroptosis in triple-negative breast cancer (TNBC).Methods:TNBC cell line MDA-MB-231 was used to establish dCK knockdown and different phosphorylation phenotypes cell models, and were treated with ferroptosis activator Erastin and / or ferroptosis inhibitor ferrostatin-1 (Fer-1) combined with / without X-ray irradiation. Cell viability was detected by MTT assay. The level of reactive oxygen species (ROS) was measured by 2′,7′-dichlorofluorescin diacetate (DCFH-DA) staining. The expression levels of dCK, transferrin, transferrin receptor (TfR1), ferroportin (FPN) and ferritin heavy chain 1 (FTH1) were detected by Western blot. Statistical analysis was performed by SPSS 17.0 and Origin 2021 software. Measurement data with normal distribution were expressed by Mean ±SD. The comparison between two groups was conducted by Student t-test. The comparison among three or more groups was performed by one-way analysis of variance. Results:In MDA-MB-231 cells, IR induced cell death was observed and Erastin significantly promoted radiation induced cell death, while Fer-1 was able to reverse radiation induced cell death. Compared with the control group, IR induced cell death was increased, the level of ROS was suppressed in the dCK knockdown group. Erastin combined with IR induced reduced cell death and weakened ROS level. Fer-1 reduced the degree of IR induced cell death, and it could not inhibit the induction of ROS by IR.Compared with the control cells, the rate of cell death was decreased induced by IR, the level of ROS was decreased, the expression of FTH1 was down-regulated after IR in the dCK wild-type (dCK-WT ) or dCK hyperphosphorylated (dCK-S74E) MDA-MB-231 cells. In addition, Erastin promotes IR induced cell death and increased ROS levels, while Fer-1 significantly enhances the degree of reversal of IR induced ROS and cell death in these cells. Conclusions:dCK phosphorylation increases ferroptosis induced by IR in TNBC cells. Targeting dCK may be a novel therapeutic approach to overcome radioresistance in TNBC treatment.

Application of Ultrasound Biomicroscopy in Longterm Follow-up Post Modified CO / 中国医学科学院学报

Objective To observe the role of ultrasound biomicroscopy(UBM)in two-year post-operative follow-up for primary open-angle glaucoma patients with modified CO

Effects of natural organic soil amendments on quality of Panax notoginseng / 中国中药杂志

The effects of four natural organic soil amendments on the quality and pesticide residues of Panax notoginseng were investigated through field experiments and the suitable dosage ratio of each soil amendment was selected to provide a new idea for the pollution-free cultivation of P. notoginseng. The four natural organic soil amendments used in this study were Jishibao, Jihuo, Fudujing, and omnipotent nutrients, which were produced by mixed fermentation of aboveground parts of different plants, biological waste residue, and biochar. During the experiments, only four soil amendments were applied to P. notoginseng instead of any pesticides and fertilizers. The experiment was designed as four factors and three levels. There were three dosage gradients(low, medium, and high) for Jishibao(A), Jihuo(B), Fudujing(C), and omnipotent nutrients(D). When the dosage of one soil amendment changed, the do-sage of the other soil amendments remained medium. There were 10 groups in addition to the soil amendment-free group as control(CK). The results showed that the four soil amendments could significantly improve the growth environment of P. notoginseng and increase the seedling survival rate and saponin content of P. notoginseng. The seedling survival rates of the treatment groups increased by 8.24%-30.05% as compared with the control group. Furthermore, the content of pesticide residues in P. notoginseng was too low to be detected, and that of heavy metals in P. notoginseng was far lower than the specified content in the Chinese Pharmacopoeia(2020). The optimal effect was achieved at medium dosage for all the soil amendments with the highest content of saponins, high seedling survival rate, and significantly reduced heavy metals, such as lead, cadmium, arsenic, and mercury.

Opposite Expression of HNRNPA2B1 in Cancer and Senescent Cells and Its Regulatory Role in Senescence of Cancer Cells / 中国生物化学与分子生物学报

Splicing factor Heterogeneous Nuclear Ribonucleoprotein A2/B1 (HNRNPA2B1 ) is associated with mouse lifespan and human longevity.It also plays a causal role in cancer development.However, whether it participates in cellular senescence, a biological process that contributes to individual aging and inhibits cancer, remains unknown.Here, we report that HNRNPA2B1 showed significantly increased expression in various cancer types while consistently decreased expression in multiple cellular senescence models.Knocking down HNRNPA2B1 in cancer cells leads to a series of senescence- associated phenotvpes.In line with its function as a splicing factor, HNRNPA2B1 downregulation causes alternative splicing changes in over one thousand genes, including those known to have a causal role in senescence.Our results also suggests that the E2F transcription factor 1 (E2F1 ) could regulate the expression of HNRNPA2BI, and E2F1-HNRNPA2B1 may be a new regulatory axis functioning in both cancer and cellular senescence, which might also have potential medical implications for cancer therapies.

SMYD3-PARP16 axis accelerates unfolded protein response and mediates neointima formation

Neointimal hyperplasia after vascular injury is a representative complication of restenosis. Endoplasmic reticulum (ER) stress-induced unfolded protein response (UPR) is involved in the pathogenesis of vascular intimal hyperplasia. PARP16, a member of the poly(ADP-ribose) polymerases family, is correlated with the nuclear envelope and the ER. Here, we found that PERK and IRE1

Analysis of Clinical Efficacy of Recombinant Human Brain Natriuretic Peptide Combined with Levosimendan in the Treatment of Acute Decompensated Heart Failure Complicated with Renal Insufficiency / 中国药房

OBJECTIVE：To observe the clinical effect and safety of recombinant human brain natriuretic peptide （rhBNP） combined with levosimendan in the treatment of acute decompensated heart failure （ADHF）complicated with renal insufficiency. METHODS：A total of 156 patients with ADHF complicated with renal insufficiency admitted to the Dept. of Cardiology in the Affiliated Hospital of Southwest Medical University during Jan.-Dec. 2019 were randomly divided into rhBNP group ，levosimendan group and combination group ，with 52 patients in each group. All patients received rountine treatment. On this basis ，rhBNP group was given rhBNP for injection [after 1.5 μg/kg intravenous pulse injection，intravenous dripping for 24 h with 0.007 5 μg（/ kg· min）]；leosimendan group was given Leosimendan injection 12.5 mg [intravenous dripping for 1 h with 6-12 μg（/ kg·min），then intravenous dripping for 23 h with 0.1 μg（/ kg·min）]. Combination group received drug combination according to the administration method of single drug group. Three groups received treatment for consecutive 7 d. Cardiac function indexes [heart rate （HR），left ventricular ejection fraction （LVEF），left ventricular end-diastolic diameter （LVEDD）]，mean arterial pressure （MAP），pulmonary capillary pressure （PCWP），renal function indexes [estimated glomerular filtration rate （eGFR），serum creatinine （Scr）]，serum levels of cystatin C （Cys-c）and amino-terminal brain natriuretic peptide precursor （NT-proBNP）were observed in 3 groups before and after treatment. Clinical efficacy and the occurrence of ADR were recorded. RESULTS ：Three cases withdrew from the study in rhBNP group and 1 case in levosimendan group ；152 cases completed the study. Before treatment ，there was no statistical significance in cardiac function indexes ，MAP，PCMP，renal function indexes or serum levels of Cys-C and NT-proBNP among 3 groups（P＞0.05）. After treatment ，the HP ，MAP，PCWP and serum level of NT-proBNP in 3 group as well as serum level of Cys-C in combination group were decreased significantly （P＜0.05）；the LVEF in 3 group as well as the eGFR and Scr level in levosimendan group and combination group were significantly increased （P＜0.05），compared with before treatment ；above indexes of combination group were significantly better than those of rhBNP group and levosimendan group （P＜0.05）. Total effective rate of combination group was 94.23％ ，which was significantly higher than those of rhBNP group （77.55％）and levosimendan group （76.47％）（P＜0.05）. There was no significant difference in the incidence of ADR among 3 groups（P＞ 0.05）. CONCLUSIONS ：rhBNP combined with levosimendan in the treatment of ADHF complicated with renal insufficiency can significantly increase the clinical efficacy ，and improve cardiac and renal function but don ’t increase the incidence of ADR.

The dilemma and breakthrough of oral administration of insulin / 药学学报

Diabetes is characterized by hyperglycemia, resulting from insulin deficiency or resistance, or both. Insulin plays an irreplaceable role in the treatment of diabetes. Subcutaneous injection is the main route of insulin administration, but usually leads to poor compliance and many side effects. Oral insulin is safer and more convenient, which has always been the Holy Grail for people to explore. After oral administration, insulin is absorbed into the hepatic portal vein and transported to the liver, which can activate the normal physiological functions and reduce the risk of hypoglycemia, insulin resistance, and improve patient compliance. However, the gastrointestinal tract has multiple absorption barriers such as chemical barrier, enzyme barrier, and permeation barrier. Due to the physical and chemical properties of insulin, it is difficult to achieve desired oral bioavailability. This article reviews the recent attempts and progress in the field of oral administration of insulin driven by innovative drug delivery technologies and biomaterials, including structural modification, enzyme inhibitors, absorption enhancers, various nanoparticles, liposomes, microspheres, and even microorganisms. Some clinical researches on oral insulin are also introduced.

Cluster analysis of Dendrobium officinale from different populations through agronomy characterization / 中国中药杂志

Through resources investigation and sample collection,a total number of 392 Dendrobium officinale from 38 different populations,9 provinces were processed for measuring and observing.Fourteen agronomy characterizations like stem height,stem diameter,number of node in stem were selected for further classification.The cluster analysis was performed using Ward and Euclidean method.The results showed that the threshold of genetic distance was 2.77.Thirtyeight populations were divided into 6 groups.The factor analysis showed that,the leaf shape,stem shape,pitch length and leaf color were very important factors for classification.This study establish the foundation for analyzing the genetic relationship of D.officinale from different populations.

Incidence and risk factor analysis of deep venous thrombosis in patients with severe traumatic brain injury / 中华危重病急救医学

Objective? To?estimate?the?incidence?and?risk?factors?for?deep?venous?thrombosis?(DVT)?in?patients?with?severe?traumatic?brain?injury?(TBI)?treated?in?the?intensive?care?unit?(ICU).? Methods? 105?patients?with?TBI?admitted?to?the?First?Affiliated?Hospital?of?University?of?Science?and?Technology?of?China?from?January?2016?to??June?2018?were?enrolled?[Glasgow?coma?scale?(GCS)?3-8;?concise?injury?score?for?other?parts≤3].?All?patients?did?not?receive?any?medication?or?physical?measures?to?prevent?DVT?during?hospitalization.?Bilateral?compression?Doppler?ultrasounds?of?the?double?lower?limbs?and?upper?limbs?were?performed?to?clarify?the?occurrence?of?DVT?on?the?first?day?of?admission?and?twice?a?week?until?ICU?discharge?or?the?death?of?patient.?The?examination?was?performed?by?a?senior?ultrasound?doctor.?It?was?defined?as?DVT?as?long?as?any?deep?vein?had?thrombosis.?Patients?were?divided?into?two?groups?according?to?whether?DVT?occurred?or?not?during?hospitalization.?Clinical?data?such?as?body?mass?index?(BMI),?coagulation?index,?platelet?count?(PLT)?and?deep?venous?catheterization?were?obtained?from?the?clinical?chemistry?laboratory?database?and?patient?files.?Logistic?regression?was?used?to?analyze?the?risk?factors?of?DVT.?Binary?Logistic?regression?was?used?to?calculate?the?predictive?probability?of?risk?factors.?The?predictive?value?of?risk?factors?and?predictive?probability?to?the?occurrence?of?DVT?was?analyzed?by?receiver?operating?characteristic?(ROC)?curve.? Results? In?105?patients?with?simple?TBI,?42?patients?developed?DVT?during?hospitalization,?and? the?incidence?of?DVT?was?40%.?Univariate?Logistic?regression?showed?that?high?BMI?[odds?ratio?(OR)?=?1.490,?95%?confidence?interval?(95%CI)?=?1.174-1.891,?P?=?0.001],?high?PLT?(OR?=?1.023,?95%CI =?1.006-1.040,?P =?0.007),?shorten?activated?partial?thromboplastin?time?(APTT;?OR?=?0.913,?95%CI?=?0.853-0.978,?P?=?0.010)?and?catheterization?in?deep?vein?(OR?=?0.114,?95%CI =?0.026-0.493,?P?=?0.004)?were?risk?factors?for?DVT.?It?was?shown?by?multivariate?regression?analysis?that?high?BMI?(OR?=?1.378,?95%CI =?1.411-1.665,?P?=?0.001),?high?PLT?(OR?=?1.017,?95%CI =?1.003-1.032,?P?=?0.020),?low?APTT?(OR =?0.920,?95%CI =?0.860-0.982,?P?=?0.012)?and?catheterization?in?deep?vein??(OR?=?0.113,?95%CI =?0.029-0.443,?P?=?0.002)?were?independent?risk?factors?for?DVT.?The?predictive?probability?formula:?Logit?P?=?-4.673+0.321×BMI－0.083×APTT+0.017×PLT－2.181×catheterization?in?deep?vein.?It?was?shown?by?ROC?curve?analysis?that?high?BMI,?high?PLT,?low?APTT?and?catheterization?in?deep?vein?could?predict?the?occurrence?of?DVT?in?severe?TBI?patients,?and?the?area?under?ROC?curve?(AUC)?was?0.775,?0.709,?0.709?and?0.680,?respectively.?The?AUC?of?prediction?probability?was?0.890,?and?its?sensitivity?and?specificity?were?respectively?increased?to?88.10%?and?85.71%.? Conclusions? The?incidence?of?DVT?was?higher?in?patients?with?simple?severe?TBI.?High?coagulation,?high?BMI,?high?PLT?and?catheterization?in?deep?vein?are?the?independent?risk?factors?for?DVT.

Establishment of DNA Fingerprints and Genetic Similarities Analysis of Dendrobium officinale / 中国实验方剂学杂志

This study adopted the 4 pairs of simple sequence repeats(SSR) primers selected by National Resource Center for Chinese Meteria Medica (CACMS) to detect 29 populations of Dendrobium officinale samples from 9 main places of production; 5,7,4 and 3 polymorphism bands were amplified from these 4 pairs of SSR primers. The DNA identities of different populations were constructed by SSR. The 29 D. officinale populations could be divided into 4 classes. The clustering result was related to the places of production. Samples from Yunnan,Guizhou,Sichuan provinces were classified into one category,while samples from Anhui and Guangxi provinces were classified into another category. Samples from Guangdong Danxia,Zhejiang Yongkang,Zhejiang Leqing and Taining belonged to a category. PopGene (version 1.32) software was applied to calculate the genetic similarity of the 29 D. officinale populations. The similarities was between 0.403 4 and 1.0.Based on the genetic similarity,the genetic consistency included three classes,A,B and C. Samples with a similar geographical location and landform environment have higher genetic similarities,which indicate the same genetic background. This paper provides reference information to study the identification, selection and breeding of good varieties.

HPLC Characteristic Spectrum Optimization of Flavonoid Glycosides on Dendrobium officinale and Characteristics Analysis of Different Provenances / 中国实验方剂学杂志

Objective: To optimize the HPLC characteristic spectrum of flavonoid glycosides of Dendrobium officinale,and identify the common and specific components of different provenances. Method: Kromasil 100-5 C18 column was adopted, with tetrahydrofuran-acetonitrile-methanol (10:22:5)-0.05% phosphoric acid as mobile phase (gradient elution). The detection wavelength was 340 nm,the column temperature was 30℃,and the flow rate was 1.0 mL ·min-1. Result: 13 flavonoid characteristic peaks were marked in 27 batches of D. officinale,and 7 characteristic peaks of 6 flavonoid C-glycosides (vicenin Ⅱ,vicenin Ⅰ,schaftoside,isoschaftoside,violanthin and isoviolanthin) and one flavonoid O-glycosides (rutin) was identified. 7-11 characteristic peaks were detected in different batches of samples. Among them,vicenin Ⅱ was a relatively stable common peak in different source samples,and the characteristic peaks of rutin,schaftoside and isoschaftoside were quite different. According to the relative abundance of the characteristic peaks,the samples could be divided into three categories. Among them,the first category had 10 batches of samples,which mainly came from Danxia landforms of Guangdong,Jiangxi,Fujian and Zhejiang (Wuyi) Province (which called "Danxia landform species") and characterized by detection of obvious peak of rutin. The second category had 11 batches of samples,which mainly came from Yunnan and Guangxi Province (which included "Yunnan Guangnan species" and "Guangxi Tiepilan species") and characterized by detection of violanthin and isoviolanthin. And the third category had 6 batches of samples, which were mainly derived from Zhejiang Province (which called "native species from Zhejiang") and characterized by detection of different degrees of rutin peak, but it was difficult to detect the characteristic peaks of violanthin and isoviolanthin. HPLC characteristic chromatograms of D. officinale in bionics wild cultivation and greenhouse of "Danxia landform species" and "Guangxi Tiepilan species" were compared. The results showed that the characteristic peaks in D. officinale planted in greenhouse could be detected stably,which verified the reliability of the source in D. officinale. Conclusion: The analytical method has a better separation effect on flavonoids of D. officinale, with a good reproducibility. The commonness and specificities of flavonoid glycosides components of D. officinale from different categories have basically confirmed. This suggests that Vicenin Ⅱ is suitable to be a reference peak for characteristic chromatogram. Both the relative abundance of rutin and the detection or relative abundance of violanthin and isoviolanthin peaks could be used as a reference to judge the categories of D. officinale in "Danxia landform species" or "Tiepilan species from Yunnan, South Guangdong and Guangxi" or "native species from Zhejiang".

Isolation and Identification of Main Flavonoid Glycosides of Dendrobium officinale from Danxia Species and Yunnan Guangnan Species / 中国实验方剂学杂志

Objective: To extract,isolate,purify and identify the structures of the flavonoid glycoside in Dendrobium officinale from two different origin places (Danxia species and Yunnan Guangnan species),and provide experimental reference for confirming the common flavonoid glycoside components in D. officinale. Method: ① 70% ethanol was applied to extract the total flavonoids in leaves of D. officinale from two different species. Organic solvents petroleum ether,acetic ether and water saturated n-butyl alcohol were used in turn to extract the crude extraction. Then AB-8 Macroporous resin,Sephadex LH-20 and ODS chromatographic column were applied to isolate and purify the water saturated n-butyl alcohol extraction fraction. The structures of flavonoid glycoside were identified by studying physicochemical property,applying modern spectroscopy method like HPLC,ESI-MSn,1H-NMR,13 C-NMR,etc. ② HPLC characteristic spectrum technique was used to analyse and compare the common flavonoid glycoside components in Dendrobium officinale from different origin places (Danxia species,Yunnan Guangnan species,Guangxi Tiepilan species and Zhejiang native species). Result: Five flavonoid glycoside compounds were isolated from the crude extractions of the leaves of D. officinale from two different species,and they were identified as rutin,vicenin Ⅱ,viceninⅠ,violanthin and isoviolanthin. The characteristic spectrum of vicenin Ⅱ and viceninⅠwere detected in stems of D. officinale from four different origin places (Danxia species,Yunnan Guangnan species,Guangxi Tiepilan species and Zhejiang native species),and vicenin Ⅱ had a better separation degree in the characteristic spectrum. However,the characteristic spectrum of violanthin and isoviolanthin were more obvious in Yunnan Guangnan species and Guangxi Tiepilan species,while rutin was obvious in the Danxia species. Conclusion: Vicenin Ⅱis the common flavonoid glycosides component in D. officinale from different origin places (Danxia species,Yunnan Guangnan species,Guangxi Tiepilan species and Zhejiang native species),and can be used as the internal reference material for the characteristic spectrum of D. officinale.

Content Comparison of Mannose,Glucose and Narinhenin in Dendrobium officinale and Dendrobium huoshanense / 中国实验方剂学杂志

Objective: To optimize the pre-column derivation high performance liquid chromatography (HPLC) content determination method of D-mannose and D-glucose as well as the content determination method of narinhenin in Dendrobium officinale and D. huoshanense, and compare the contents of D-mannose,D-glucose and narinhenin between D. officinale and D. huoshanense. Method: A pre-column derivation HPLC method modified by Chinese Pharmacopoeia(Ch.P) 2015 was used to simultaneously determine the contents of D-mannose and D-glucose,with acetonitrile-0.02 mol·L-1 ammonium acetate solution as mobile phase for gradient elution. Kromasil 100-5 C18 was performed with the wavelength set at 250 nm,and the flow rate was 1 mL·min-1;column temperature was 30℃. HPLC content determination of narinhenin was performed on Kromasil 100-5 C18 with the acetonitrile-methanol-0.4% phosphoric acid solution as mobile phase for gradient elution,and the wavelength was set at 290 nm; the flow rate was 0.8 mL·min-1,and column temperature was 40℃. Result: D-mannose and D-glucose showed a good linear relationship within the range of 0.15-3.0 μg and 0.075-2.25 μg (r=0.999 9); and their average recoveries were 99.01% (RSD 2.1%) and 101.69% (RSD 2.0%) respectively. In addition, the other methodological researches such as repeatability and durability all met the requirements. The contents of D-mannose(Cm),D-glucose(Cg) and sum of them (Cm+Cg) were 12.75%-36.40%,2.93%-18.39% and 19.23%-54.58% in 43 batch of D. officinale. Almost all of the results except very few samples reached the D-mannose standard in Ch.P 2015, and the total content of D-mannose and D-glucose was also up to the total polysccharide standard in Ch.P. The correlation between content and origin was not significant. The contents of D-mannose(Cm),D-glucose(Cg) and sum of them (Cm+Cg) were 14.33%-29.47%,6.64%-15.20%,and 25.73%-44.37% in 12 batch of D. huoshanense. These contents and ratio of peak areas of D-mannose to D-glucose (Am/Ag) were within the scope of D. officinale's; in addition, their average contents were basically the same with those in D. officinale (about 33%).Next,naringenin showed a good linear relationship within the range of 0.020 8-0.832 0 μg (r=0.999 9),and its average recovery was 101.96% (RSD 1.8%). The content of naringenin was 0.053 2-0.122 4 mg·g-1 (average value of 0.081 0 mg·g-1) in 11 batch of D. officinale, slightly higher than 0.040 3-0.090 0 mg ·g-1 (average value of 0.068 3 mg ·g-1) in 7 batch of D. huoshanense. All of these results of narinfenin did not reach the content lower limit in Ch.P. Conclusion: The method used to determinate the content of D-mannose and D-glucose is reproducible, and their sum content is possible to substitute the total polysccaride determination (with higher errors) in D. officinale; monosaccharide content determination can be used for quantitative quality control of D. huoshanense. However, it could not distinguish D. officinale and D. huoshanense by determining the contents of polysccharide,D-glucose,D-mannose and narinhenin, and shall be combined with other specificity methods for further identification.

Antioxidation Activities in Vitro of Vicenin Ⅱ Isolated from Dendrobii Officinalis Caulis and Effect on HepG2 Cells / 中国实验方剂学杂志

Objective: To study the antioxidation activities in vitro of a comment flavonoid component named vicenin Ⅱ(Apigenin 6,8-di-C-glucoside) in Dendrobii Officinalis Caulis from different origin places and investigate its effects on apoptosis of HepG2 cells. Method: The antioxidation activities in vitro of vicenin Ⅱ (0.005-1 g·L-1) were evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH), salicylic acid and 2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid(ABTS) and copper ion reduction assays. Methye thiazolye telrazlium(MTT) assay was used to test the inhibitory effect of vicenin Ⅱ(12.5~100 μmol·L-1) on proliferation of 6 tumour cells in vitro. In subsequent apoptosis experiment, the concentration of vicenin Ⅱ was 75 μmol·L-1. The morphological changes of HepG2 cells were evaluated by Hoechst 33258 under fluorescence microscope; and the cell apoptosis rate was detected by flow cytometry with AnnexinV/PI apoptosis assay kit. The mRNA expressions of mitogen activated protein kinase (MAPK) pathway related apoptotic genes were detected by Real-time PCR assay. Result: The 1 g·L-1 vicenin Ⅱ showed 48.82% and 22.01% for DPPH scavenging rate and Cu2+ reduction rate respectively(P-1 vicenin Ⅱ showed 86.88% for ABTS scavenging rate(P-1 Vicenin Ⅱ, the cells survival rate was 45.69%(PPN-terminal kinase (JNK), and nuclear transcription factor (NF)-κB were increased(PConclusion: The general flavone glycosides component vicenin Ⅱ of Dendrobii Officinalis Caulis from different origins has a certain antioxidation effect and significant inhibitory effect on proliferation, and could induce apoptosis on HepG2 cells probably by regulating the expression of related genes in MAPK pathway and Bax/Bcl-2.

Recent progress in drug delivery

Drug delivery systems (DDS) are defined as methods by which drugs are delivered to desired tissues, organs, cells and subcellular organs for drug release and absorption through a variety of drug carriers. Its usual purpose to improve the pharmacological activities of therapeutic drugs and to overcome problems such as limited solubility, drug aggregation, low bioavailability, poor biodistribution, lack of selectivity, or to reduce the side effects of therapeutic drugs. During 2015-2018, significant progress in the research on drug delivery systems has been achieved along with advances in related fields, such as pharmaceutical sciences, material sciences and biomedical sciences. This review provides a concise overview of current progress in this research area through its focus on the delivery strategies, construction techniques and specific examples. It is a valuable reference for pharmaceutical scientists who want to learn more about the design of drug delivery systems.