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.

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

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.

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

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.

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.

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.

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.

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".

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.

Effects of Atorvastatin Combined with CORM- 3 on Inflammation and Oxidative Stress Indexes in Atherosclerotic Vulnerable Plaque Model Rats / 中国药房

OBJECTIVE： To study the effects of atorvastatin combined with carbon monoxide releasing molecule 3 （CORM-3） on inflammation and oxidative stress indexes in atherosclerotic （AS） vulnerable plaque model rats. METHODS： The rats were randomly divided into control group （normal saline， i.g.）， model group （normal saline， i.g.）， statin group （atorvastatin 2 mg/kg，  i.g.）， and statin+CORM-3 group （atorvastatin 2 mg/kg， i.g.+CORM-3 10 mg/kg， i.p.）， with 8 rats in each group. Control group was fed with basal diet， and the right common carotid artery was exposed to surgery without injury and was treated with normal saline instead of drug； other three groups were fed with high-fat diet+right common carotid artery injury+heteroprotein injection to induce AS vulnerable plaque model， for 10 weeks； and then they were given relevant medicine for intervention， once a day， for consecutive 2 weeks. 24 h after last medication， abdominal artery blood was collected； the concentration of LDL-C and HDL-C were determined by fully automatic biochemical analyzer. The levels of hs-CRP， IL-10， MCP-1 and MMP-9 in plasma were detected by ELISA； plasma levels of MDA and oxidized low density lipoprotein （ox-LDL） were determined by chemical colorimetry； the protein expression of heme oxygenase-1 （HO-1） was determined by Western blot. The pathological changes of right common carotid artery were observed under light microscope. RESULTS： Compared with control group， the levels of LDL-C， hs-CRP， MCP-1， MMP-9， MDA and ox-LDL， and protein expression of HO-1 were increased significantly （P＜0.05）， while the levels of HDL-C and IL-10 were decreased significantly in model group （P＜0.05）； the right common carotid artery formed obvious AS plaques. Compared with model group， the levels of LDL-C， hs-CRP， MCP-1， MMP-9， MDA and ox-LDL were decreased significantly in statin group and statin+CORM-3 group in model group （P＜0.05）， while the levels of HDL-C， IL-10 and the protein expression of HO-1 were increased significantly （P＜0.05）. Except for LDL-C and HDL-C， the improvement of other indexes in statin+CORM-3 group was more significant than statin group （P＜0.05）； pathological changes of right common carotid artery in statin group were not obvious， but the pathological changes of rats in statin+CORM-3 group were significantly alleviated and plaque structure also tended to be more stable. CONCLUSIONS： Atorvastatin combined with CORM-3 is better than atorvastatin alone in improving inflammation and oxidative stress indexes of AS vulnerable plaque model rats， and can promote the stability of AS vulnerable plaques.

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.

Gene therapy and delivery strategies for ocular diseases / 药学学报

Gene therapy has obvious advantages in the treatment of ocular diseases due to the unique structure of the eye. In recent years, there are more and more therapeutic gene-based drugs for ophthalmic application in clinical trials. Most of the delivery vectors are adeno-associated virus and administered via intraocular injection, which has potential risks. Traditional remedies, such as topical instillationor systemic administration, have limited therapeutic effects on the diseases in the posterior segment of the eye, where the chemical drugs are hard to reach. This makes the research of new strategies for gene drug delivery extremely urgent. For better understanding of the latest hot topics of ocular gene therapy, this article is prepared to introduce application of gene therapy to the typical ocular diseases and the corresponding gene-based medicines. The absorption routes for gene delivery into eyes and existing barriers are summarized. Finally, the gene delivery strategies are highlighted. The clinical application of ocular gene therapy will be boosted by overcoming the absorbing barriers and reducing the potential pitfalls.

Development and application of competitive light-initiated chemiluminescent assay for quantitation of total IgE in human serum / 临床检验杂志

Objective To develop a competitive immunoassay for quantitative determination of total immunoglobin E (tIgE) in human serum based on light-initiated chemiluminescent assay (LICA).Methods The LICA-tIgE assay was performed by incubating serum samples or calibrator with anti-human IgE antibody-coated chemiluminescet beads,biotinylated human IgE and streptavidin-coated sensitizer beads.The working conditions of this assay were optimized,analytical performance was detected and the correlation of tIgE results between LICA and Beckman Coulter IMMAGE 800 was evaluated.Results The precision of intra-assay and inter-assay (coefficient of variation) ranged from 5.50％ to 7.73％ and 6.45％ to 9.90％,respectively.The functional sensitivity of this assay was 12.65 IU/mL.The recovery rates measured by adding IgE calibrators to human sera with different IgE concentrations were ranged from 104.15％ to 109.37％.The disturbing rates measured by adding total bilirubin,hemoglobin and triacylglycerol to human sera with different IgE concentrations were ranged from-4.49％ to 8.46％.Also,the tIgE results of 111 patients measured by LICA correlated well with those by Beckman Coulter IMMAGE 800 (r2 =0.959).Conclusion LICA developed in this study for detecting tIgE of human serum showed effective perfomance and could meet the basic requirements of clinical diagnostic reagents.

Effect of Qigui Yishen Decoction on miR-141 regulation in UUO mice with renal fibrosis / 中成药

AIM To investigate the effect and mechanism of Qigui Yishen Decoction (QGYS,Astragali Radix,Angelicae sinensis Radix,Chuanxiong Rhizoma,Achyranthis bidentatae Radix) on regulating the expression of miR-141 in unilateral ureteral obstruction (UUO) mice with renal fibrosis.METHODS Thirty Balb/c male mice randomly divided into sham-operated group (n =6),UUO group (n =6),Lotensin (50 g/kg) group (n =6),QGYS high dose (50 g/kg) group (n =6),and QGYS low dose (10 g/kg) group (n =6) were conducted UUO surgery to promote kidney fibrosis except the six mice in the sham operation group.After a successive 10-day medication of QGYS and Lotensin to mice by oral gavage on daily basis,all mice were killed to procure renal tissue to observe its morphology and pathology changes by HE staining.The expressions of TGF-β1,ColⅣ,and MMP-9 were analyzed by immunohistochemical method,and the expressions of miR141,TGF-β1 were measured by real-time PCR.RESULTS The obviously pathological injuries including renal interstitial fibrosis were identified by HE staining among the groups intervened with UUO,but the variance in the extent due to different administrations of QGYS and Lotensin was noticed as well (P ＜ 0.05).As compared to the UUO group,high and low dose QGYS groups and Lotensin group achieved an up-regulated expression of TGF-β1 and ColⅣ,and a down-regulated expression of MMP-9 by immunohistochemistry (P ＜ 0.05),and significantly increased Mrna expression of miR-141,and decreased Mrna expression of TGF-β1 by real-time PCR (P ＜ 0.05).CONCLUSION In UUO mouse models,QGYS gives influence to TGF-β1and MMP-9 through inducing miR-141 expression change to decrease abnormal accumulation of ECM,and thus inhibits the progression of renal fibrosis.

Antisense transcription regulates the expression of sense gene via alternative polyadenylation

Natural antisense transcripts (NAT) and alternative polyadenylation (APA) of messenger RNA (mRNA) are important contributors of transcriptome complexity, each playing a critical role in multiple biological processes. However, whether they have crosstalk and function collaboratively is unclear. We discovered that APA enriched in human sense-antisense (S-AS) gene pairs, and finally focused on RNASEH2C-KAT5 S-AS pair for further study. In cis but not in trans over-expression of the antisense KAT5 gene promoted the usage of distal polyA (pA) site in sense gene RNASEH2C, which generated longer 3' untranslated region (3'UTR) and produced less protein, accompanying with slowed cell growth. Mechanistically, elevated Pol II occupancy coupled with SRSF3 could explain the higher usage of distal pA site. Finally, NAT-mediated downregulation of sense gene's protein level in RNASEH2C-KAT5 pair was specific for human rather than mouse, which lacks the distal pA site of RNASEH2C. We provided the first evidence to support that certain gene affected phenotype may not by the protein of its own, but by affecting the expression of its overlapped gene through APA, implying an unexpected view for understanding the link between genotype and phenotype.

Differential stem cell aging kinetics in Hutchinson-Gilford progeria syndrome and Werner syndrome

Hutchinson-Gilford progeria syndrome (HGPS) and Werner syndrome (WS) are two of the best characterized human progeroid syndromes. HGPS is caused by a point mutation in lamin A (LMNA) gene, resulting in the production of a truncated protein product-progerin. WS is caused by mutations in WRN gene, encoding a loss-of-function RecQ DNA helicase. Here, by gene editing we created isogenic human embryonic stem cells (ESCs) with heterozygous (G608G/+) or homozygous (G608G/G608G) LMNA mutation and biallelic WRN knockout, for modeling HGPS and WS pathogenesis, respectively. While ESCs and endothelial cells (ECs) did not present any features of premature senescence, HGPS- and WS-mesenchymal stem cells (MSCs) showed aging-associated phenotypes with different kinetics. WS-MSCs had early-onset mild premature aging phenotypes while HGPS-MSCs exhibited late-onset acute premature aging characterisitcs. Taken together, our study compares and contrasts the distinct pathologies underpinning the two premature aging disorders, and provides reliable stem-cell based models to identify new therapeutic strategies for pathological and physiological aging.