Study on quality control method of the roots and rhizoma of Toricellia angulata / 中国药房

OBJECTIVE To establish the quality control method for the roots and rhizoma of Toricellia angulata. METHODS The properties of the roots and rhizoma of T. angulata were observed and microscopic identification was conducted. The moisture， total ash， acid-insoluble ash and ethanol-soluble extract were examined according to the method stated in the 2020 edition of Chinese Pharmacopoeia （part Ⅳ）. HPLC fingerprints of 11 batches of the roots and rhizoma of T. angulata were established， common peaks were identified and the similarity was evaluated by using the Similarity Evaluation System of Chromatographic Fingerprint of TCM （2012 edition）. The contents of coniferin， syringin， chlorogenic acid， （+）-syringaresinol-O-β-D-glucopyranoside and syringaresinol were determined by HPLC. RESULTS The properties and microscopic identification of the roots and rhizoma of T. angulata were obvious. The average contents of moisture， total ash， acid-insoluble ash and ethanol-soluble extract were 7.54%， 2.18%， 0.15% and 7.81%， respectively. There were 16 common peaks marked in the HPLC fingerprints of 11 batches of the roots and rhizoma of T. angulata， with similarities of 0.856-0.960； five of them were identified， such as coniferin， syringin， chlorogenic acid， （+）-syringaresinol-O-β-D-glucopyranoside and syringaresinol. The contents of the above five components were 0.047 2-0.401 6， 0.836 8-8.697 9， 1.245 3-10.950 0， 0.139 0-0.437 8 and 0.016 4-0.635 3 mg/g， respectively. CONCLUSIONS The established method is stable and accurate， which can be used for the quality control of the roots and rhizoma of T. angulata. It is preliminarily proposed that the moisture in the roots and rhizoma of T. angulata is not more than 11.0%， the total ash is not more than 4.0%， the ethanol-soluble extract is not less than 5.0%， the contents of coniferin， syringin， chlorogenic acid， （+）-syringaresinol-O-β-D- glucopyranoside and syringaresinol are not less than 0.04，0.83， 1.24， 0.13， 0.01 mg/g， respectively.

Mechanism of ginkgo flavonoid aglycone against doxorubicin-induced cardiotoxicity / 中国药房

OBJECTIVE To investigate the potential mechanism of the effect of ginkgo flavone aglycone （GA） against doxorubicin （DOX）-induced cardiotoxicity. METHODS The male ICR mice were randomized into control group （CON group）， model group （DOX group） and GA+DOX group （GDOX group）， with 12 mice in each group. The DOX group was injected with DOX solution at a dose of 3 mg/kg via tail vein every other day， and the GDOX group was given GA suspension intragastrically at a dose of 100 mg/kg every day+DOX solution at a dose of 3 mg/kg via tail vein every other day， for 15 consecutive days. After the end of administration， the serum levels of aspartate aminotransferase（AST）， creatine kinase（CK）， creatine kinase isoenzyme（CK- MB） and lactate dehydrogenase（LDH） in mice were detected in each group. Based on the metabolomics method， UHPLC-Q- Exactive Orbitrap HRMS method was used； based on principal component analysis （PCA） and orthogonal partial least squares- discriminant analysis （OPLS-DA）， the differentially expressed metabolites （DEMs） were screened using the criteria of variable importance in the projection≥1， fold change of peak area＞1 and P＜0.05； biological analysis was conducted based on databases such as HMDB and PubChem. RESULTS Compared with CON group， serum levels of AST， CK， CK-MB and LDH were increased significantly in DOX group （P＜0.05）； compared with DOX group， the serum levels of the above indicators （except for CK-MB） were decreased significantly in GDOX group （P＜0.05）. PCA and OPLS-DA showed that myocardial tissue samples of CON group， DOX group and GDOX group were isolated completely. After database matching， 37 common DEMs were identified， among which 17 DEMs were significantly up-regulated in the DOX group and significantly down- regulated in the GDOX group， and 8 DEMs were significantly down-regulated in the DOX group and significantly up-regulated in the GDOX group； pathway enrichment involved the biosynthesis of unsaturated fatty acids， arachidonic acid metabolism， linoleic acid metabolism， taurine and hypotaurine metabolism； the key metabolites in the above pathways included docosahexaenoic acid， arachidonic acid， phosphatidylcholine （16∶0/18∶3） and taurine. CONCLUSIONS GA may regulate the biosynthesis of unsaturated fatty acids， arachidonic acid metabolism and other metabolic pathways by acting on the core metabolites such as docosahexaenoic acid and arachidonic acid， thus alleviating the cardiotoxic effects of DOX.

Transcriptome profiling of abdominal aortic tissues reveals alterations in mRNAs of Takayasu arteritis.

Takayasu arteritis (TA) is a chronic granulomatous vasculitis involving in the main branches of aorta. Previous studies mainly used peripheral blood and some vascular tissues but seldom studies have sequenced vascular tissues. Here in this study, we aimed to explore the alterations of mRNA in TA by performing bulk RNA sequencing. A total of 14 abdominal aortic tissues including 8 from renal transplantation and 6 from patient with TA undergoing bypass surgeries. Bulk RNA sequencing were performed and after the quality control, a total of 1897 transcripts were observed to be significantly differently (p < 0.05 and Log2FC > 1) expressed between the TA and control group, among which 1,361 transcripts were in TA group and 536 in the Control group. Reactome Pathway Enrichment Comparison analysis revealed interleukin-10 signaling and signaling by interleukins were highly expressed in TA group. Besides, extracellular matrix organization was also observed in this group. WGCNA and PPI obtained 26 core genes which were highly correlated with the clinical phenotype. We then also perform deconvolution of the bulk RNA-seq data by using the scRNA-seq dataset and noticed the high proportion of smooth muscle cells in our dataset. Additionally, immunohistochemical staining confirmed our bioinformatic analysis that TA aortic tissues express high levels of IL-1R1 and IL-1R2. Briefly, this study revealed critical roles of interleukins in TA pathogenesis, and SMCs may also participate in the reconstruction in vessel wall at late stage of TA.

Cardioprotective effect of NRG-4 gene expression on spontaneous hypertension rats and its mechanism through mediating the activation of ErbB signaling pathway.

To explore the myocardial protective effect of Neuregulin-4 (NRG-4) gene expression on spontaneous hypertension (SHR) rats and its mechanism through mediating the activation of the ErbB signaling pathway, this study was conducted. For this purpose, forty 24-week-old male SPF SHR rats were selected as the experimental group, and 10 age and sex-matched Wistar Kyoto (WKY) rats were selected as the control group. Cardiac tissues were collected for hematoxylin-eosin (HE) staining, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining, Masson staining, and immunohistochemical staining. Following tail vein injection of recombinant lentiviral vectors, the experimental groups were constructed as the control group (SHR rats without any treatment), Empty vector group (Empty Vector transfection), shRNA negative control (NC) group (LV-shRNA-NRG-4 NC transfection to silence the expression of NRG-4), shRNA group (LV-shRNA-NRG-4 transfection), pcDNA3.1(-) NC group (pcDNA3.1(-)-NRG-4 empty vector transfection) and pcDNA3.1(-) group (pcDNA3.1(-)-NRG-4 transfection to overexpress NRG-4). Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expression levels In addition, methyl thiazolyl tetrazolium salt (MTT) assay and flow cytometry were performed to detect the proliferation and apoptosis of cardiomyocytes, respectively. Results showed that in the SHR group, the cardiomyocytes showed hypertrophy, disordered arrangement, hyperchromatic nucleus, irregular shape, obvious rupture of myocardial fibers, and obvious proliferation of fibrous stroma; obvious myocardial fibrosis, and there were more blue collagen fibers around cardiomyocytes and myocardial arterioles; cardiomyocytes were swollen, muscle fibers arranged disorderly, collagen around the coronary artery and myocardial interstitium increased significantly with a cross-linking appearance; besides, compared with WKY group, the apoptosis index of cardiomyocytes in SHR group was significantly increased (P<0.05). The expression of NRG-4 protein was decreased in the SHR group compared with the WKY group (P<0.05). In vitro, there was no difference in the mRNA expression of NRG-4, ErbB2 and ErbB4, MMP2, TGFß1 and α-SMA, as well as Caspase3, Bax and Bcl-2 among the control group, Empty vector group, shRNA NC group and pcDNA3.1(-) NC group (P>0.05). While shRNA group showed decreased expressions of NRG-4, ErbB2, ErbB4, MMP2, TGFß1, α-SMA and Bcl-2, while increased Caspase3 and Bax expressions, as well as promoted cell proliferation and cell apoptosis when compared with the shRNA NC group (P<0.05); while compared with pcDNA3.1(-) NC group, pcDNA3.1(-) group had highly increased expressions of NRG-4, ErbB2, ErbB4, MMP2, TGFß1, α-SMA and Bcl-2, while decreased Caspase3 and Bax expressions, inhibited cell proliferation and cell apoptosis (all P<0.05). It is concluded Upregulation of NRG-4 gene expression can promote the activation of the ErbB signaling pathway, thus inhibiting the proliferation and apoptosis of cardiomyocytes in SHR rats, reversing myocardial fibrosis, and playing its cardioprotective role.

Progress of long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 in hematological malignancies / 白血病·淋巴瘤

Metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) plays an important role in the development of various tumors. Recent studies have shown that MALAT1 is highly expressed in hematological tumors and can participate in development, progression and prognosis of hematological tumors at transcriptional and post-transcriptional levels as a competitive endogenous RNA. Therefore, MALAT1 might be a novel marker as a valuable basis for the diagnosis, treatment and prognosis evaluation of hematological tumors.

Effects of Chinese patent medicine for activating blood circulation on cardiac repair after myocardial infarction / 中国临床药理学与治疗学

Myocardial infarction (MI) is a cardiovascular disease with high morbidity and mortality, which seriously endangers human health. Poor ways of repair after MI may damage people's life quality, therefore scientists have been committed to exploring the way of myocardial repair after MI to strive for a better prognosis of these patients. Chinese patent medicines for blood circulation activation have a long history of use in treatment of MI, where they have been shown to attenuate myocardial injury after MI and promote cardiac repair with multiple targets, links and pathways. This review focuses on a few representative Chinese patent medicines for MI and summarizes their pharmacological effects and clinical research in cardiac repair following MI, effectively providing scientific foundation on treating MI with Chinese patent medicines.

Improvement of the quality standard of Gerbera piloselloides / 中国药房

OBJECTIVE To improve th e quality standard for Gerbera piloselloides. METHODS The properties of G. piloselloides were observed and microscopic identification was conducted for powder. The moisture ，total ash ，acid-insoluble ash and ethanol-soluble extract were detected according to the method stated in 2020 edition of Chinese Pharmacopoeia （part Ⅳ）. The contents of nodakenin，luteolin-7-O-β-D-lutinoside，luteoloside，apigenin-7-O-β-D-lutinoside，apigenin-7-O-β-D-glucopyranoside and marmesin were determined by high performance liquid chromatography method. RESULTS G. piloselloides were shrunken ，densely covered with thick white cotton wool ，with many fibrous roots ；the surface was taupe or gray-brown ；its texture was brittle and easy to break ；the cross section was yellow-white ，and there was an obvious small wooden heart in the center. Its powder was tan ， and non-glandular hairs ，stone cells ，calcium oxalate cubes ，ducts，fibers could be seen unde r microscope. The measured values of moisture，total ash ，acid-insoluble ash and alcohol-soluble extract ， for 15 batches of samples were 8.63％-11.34％，10.39％-14.93％， 3.29％-6.37％ and 9.03％-15.02％，respectively；average values were 10.01％，12.26％，4.61％，12.36％. The linear ranges of nodakenin，luteolin-7-O-β-D-rutinoside，luteoloside，apigenin- 7-O-β-D-lutinoside，apigenin-7-O-β-D-glucopyranoside and marmesin we re 3.87-154.88，1.64-65.41，1.60-64.00，1.92-76.96， 1.27-50.93，0.40-15.89 μg/mL，respectively（r≥0.999 1）. RSDs of precision ，stability（24 h）and repeatability tests were all less than 3％. The average recoveries were 101.88％，100.89％，102.64％，95.75％，96.71％ and 103.48％，respectively；RSDs were 0.55％，0.43％，0.34％，0.49％，0.47％ and 0.37％，respectively（n＝6）；the contents of above 6 components were 0.152 7-0.852 2， 0.084 5-0.669 7，0.136 7-0.961 0，0.126 0-1.193 2，0.128 8-1.102 2，0.046 9-0.678 0 mg/g. CONCLUSIONS The established method can be used for the quality control of G. piloselloides . It is preliminarily proposed that the moisture in G. piloselloides is not more than 12.0％；the total ash is not more than 15.0％，the acid-insoluble ash is not more than 6.0％，the alcohol-soluble extract is not less than 9.0％；the contents of luteoloside and apigenin- 7-O-β-D-glucopyranoside are not less than 0.016％.

Effects of increasing efficacy and decreasing toxicity of ginkgo flavone aglycone on doxorubicin in the treatment of liver cancer / 中国药房

OBJECTIVE To study the effects of increasing efficacy and decreasing toxicity of ginkgo flavone aglycone （GA） on doxorubicin （DOX）in the treatment of liver cancer. METHODS A tumor bearing model was established by inoculating liver cancer cell H 22 into the right axillary skin of ICR mice. The successfully modeled mice were randomly divided into model control group，DOX group （2.5 mg/kg，once every other day ，via tail vein ），GA group （30 mg/kg，once a day ，gavage）and GA+DOX group（the usage was the same as single drug groups ），with 6 mice in each group. The administration cycle was 15 days. The general growth of mice in each group were observed ，body weight and tumor weight were measured ，and the inhibition rate of tumor was calculated. Jin’s formula was used to evaluate the effect of combined medication （Q）. The serum level of alpha-fetal protein（AFP），the pathological changes of tumor tissue ，cell apoptosis and the expression of platelet-endothelial cell adhesion molecule-1（CD31）were detected in each group. The cardiac index，serum levels of B-type natriuretic peptide （BNP）and N-terminal pro-brain natriuretic peptide （NT-pro BNP ），pathological changes of heart and myocardial fibrosis degree were also detected. RESULTS The percentage of body weight change （except for GA group ） and tumor weights of DOX group，GA group and GA + DOX group were all decreased significantly，compared with model control group （P＜0.05 or P＜0.01），while tumor weight of GA+DOX gro up was significantly lower than DOX group （P＜0.01）. Inhibitory rates of tumor in 3 administration groups were 54.29％，42.50％ and 89.29％ respectively，and Q of two-drug combination was 1.21. The tumor tissues of mice in each administration group were necrotic to varying degrees ；the serum level of AFP and the expression of CD31 in tumor tissue were decreased significantly ，compared with model control group （P＜0.05 or P＜0.01）；the percentage of necrosis area of tumor tissue and the positive rate of apoptosis （except for single drug groups ）were significantly increased （P＜0.05 or P＜0.01），while positive rate of apoptosis in GA+DOX group was significantly higher than DOX group （P＜0.05）. Cardiac index of mice in DOX group was significantly lower than model control group （P＜0.01）；serum levels of BNP and NT-pro BNP in DOX group and GA+ DOX group were significantly higher than model control group （P＜0.05 or P＜0.01）；pathological changes of heart and the degree of myocardial fibrosis in GA+DOX group were lower than DOX group. CONCLUSIONS GA combined with DOX show synergistic antitumor effect. GA can strengthen the apoptosis promoting effect of DOX ，and can help to reduce the cardiotoxicity of DOX.

Single-Cell RNA Sequencing Revealed CD14+ Monocytes Increased in Patients With Takayasu's Arteritis Requiring Surgical Management.

Objectives: Takayasu Arteritis (TA) is a highly specific vascular inflammation and poses threat to patients' health. Although some patients have accepted medical treatment, their culprit lesions require surgical management (TARSM). This study aimed at dissecting the transcriptomes of peripheral blood mononuclear cells (PBMCs) in these patients and to explore potential clinical markers for TA development and progression. Methods: Peripheral blood were collected from four TA patients requiring surgical management and four age-sex matched healthy donors. Single cell RNA sequencing (scRNA-seq) was adopted to explore the transcriptomic diversity and function of their PBMCs. ELISA, qPCR, and FACS were conducted to validate the results of the analysis. Results: A total of 29918 qualified cells were included for downstream analysis. Nine major cell types were confirmed, including CD14+ monocytes, CD8+ T cells, NK cells, CD4+ T cells, B cells, CD16+ monocytes, megakaryocytes, dendritic cells and plasmacytoid dendritic cells. CD14+ monocytes (50.0 vs. 39.3%, p < 0.05) increased in TA patients, as validated by FACS results. TXNIP, AREG, THBS1, and CD163 increased in TA patients. ILs like IL-6, IL-6STP1, IL-6ST, IL-15, and IL-15RA increased in TA group. Conclusion: Transcriptome heterogeneities of PBMCs in TA patients requiring surgical management were revealed in the present study. In the patients with TA, CD14+ monocytes and gene expressions involved in oxidative stress were increased, indicating a new treatment and research direction in this field.

Establishment and validation of a model of a contrast-enhanced ultrasound scoring system for the prediction of renal artery stenosis in elderly patients / 中华老年医学杂志

Objective:To establish and validate a prognostic model of a contrast-enhanced ultrasound scoring(CEUS)system for evaluating renal artery stenosis(RAS)in the elderly.Methods:This was a single-center retrospective study.A total of 324 elderly RAS patients admitted to Beijing Hospital from October 2017 to July 2020 were randomly assigned into the model group(n=174)and the validation group(150)in a 1∶1 ratio.Clinical and imaging data of patients on admission including general conditions, previous medical history, blood pressure, blood creatinine, renal artery stenosis and cortical blood perfusion in the affected kidney and renal function(GFR)at 1-year follow-up were collected.Univariate and multivariate logistic regression was used to establish a model of the CEUS scoring system.The receiver operating characteristic(ROC)curve and area under the ROC curve(AUC)were used to evaluate prediction accuracy.Clinical application value of the CEUS scoring system model was evaluated via decision curve analysis using a nomogram.Results:Baseline clinical and radiomic data had no significant difference between the model group and the validation group( P>0.05). Multivariate logistic regression analysis results showed that age( OR=1.242, 95% CI: 1.081-1.427, P<0.01), diabetes( OR=1.545, 95% CI: 1.107-2.156, P<0.05), blood pressure( OR=1.328, 95% CI: 1.056-1.670, P<0.05), renal function( OR=2.374, 95% CI: 1.216-3.887, P<0.01)and cortical blood perfusion parameter( OR=2.646, 95% CI: 1.553-6.369, P<0.01)were risk factors for the deterioration of renal function during 1 year follow-up.Based on these results, a nomogram for the CEUS scoring system model was drawn, and its consistency index, the C-Index, was 0.725(95% CI: 0.653-0.776). The AUC of the CEUS scoring system was 0.824 and the Youden index was 0.711 in the model group, with a specificity of 0.774 and a sensitivity of 0.837.The AUC of the CEUS scoring system was 0.853 and the Youden index was 0.715 in the validation group, with a specificity of 0.684 and a sensitivity of 0.889.There was no significant difference in ROC curve between the two groups( D=1.387, P>0.05). In addition, calibration charts of the two models showed that the calibration curve of the CEUS scoring system was close to the standard curve, with no statistically significant difference( P>0.05). Conclusions:The CEUS scoring system model can be used to predict the risk of worsening renal function in elderly RAS patients during 1-year follow-up.

An observational study of the effect of baseline renal cortical blood perfusion with contrast-enhanced ultrasound on short-term outcomes of stent implantation for severe renal artery stenosis / 中华超声影像学杂志

Objective:To observe the effect of the cortical blood perfusion parameter of wash-in area under curve (iAUC) with contrast-enhanced ultrasound(CEUS) on the effect of short-term outcomes of stent implantation in patients with severe renal artery stenosis (RAS).Methods:Retrospective analysis was performed on 82 patients with unilateral severe RAS who received stent implantation in Beijing Hospital from October 2017 to December 2019. According to the baseline iAUC before CEUS, all patients were divided into the poorly-perfused group (iAUC<850.0 dB×s) (37 cases) and the well-perfused group (iAUC≥850.0 dB×s) (45 cases). Baseline and perioperative clinical-imaging data were analyzed between the two groups. Followed up for 10-12 (11.5±1.7) months, Kaplan-Meier survival curves and Log-rank test were used to analyze the rate of adverse cardiac and renal vascular events and hypertension control rates.Results:Compared with the well-perfused group, the poorly-perfused group showed a longer course of hypertension, more diabetic patients, higher systolic blood pressure, diastolic blood pressure, 24 h average systolic blood pressure, and 24 h average diastolic blood pressure, lower glomerular filtration rate, and severe renal artery stenosis. Besides, the iAUC, wash-out AUC and the peak intensity were lower, the average transit time was longer, and the hypoglycemic treatment rate was higher (all P<0.05). Kaplan-Meier survival curve and Log-rank test analysis showed that the occurrence of cardio-renal vascular events ( HR=0.361, 95% CI=0.144-0.907, P=0.012) and renal function deterioration rate ( HR=0.286, 95% CI=0.090-0.914, P=0.035) in the well-perfused group were significantly lower than those in the poorly-perfused group. The blood pressure results demonstrated that the effective rate of hypertension treatment in the well-perfused group was significantly higher than that in the poorly-perfused group (93.3% vs 59.5%, P<0.001), but the improvement rate of hypertension (60.0% vs 43.2%) and cure rate (28.9% vs 16.2%) were not statistically significant between the two groups(all P>0.05). Conclusions:Severe RAS patients with decreased baseline iAUC often have diabetes, longer duration of hypertension, significantly reduced glomerular filtration rate and more severe RAS, short-term outcomes are worse with stent implantation.

Progress of the mechanism of Wnt signaling pathway in T-cell acute lymphoblastic leukemia / 白血病·淋巴瘤

T-cell acute lymphoblastic leukemia (T-ALL) is derived from the malignant transformation and clonal proliferation of precursor T cells. T-ALL is usually associated with the genetic mutations and/or chromosomal translocation, thus causing the change of survival, proliferation and progenitor cell differentiation of T cells in T-cell development. Studies have shown that Wnt signaling pathway plays a key role in the regulation of hematopoietic stem cell and normal T-cell development, and it is also abnormally altered in T-ALL. This article reviews the research progress of the mechanism of Wnt signaling pathway in T-ALL development.

Remote endarterectomy for complex lower extremity ischemia / 中华普通外科杂志

Objective:To evaluate the effect of remote endarterectomy in the treatment of complex lower extremity ischemia.Methods:Twenty-one limb ischemic patients underwent remote endarterectomy in Beijing Hospital from Sep 2016 to Feb 2020. Clinical data including general condition, the lesion of lower artery before operation and follow up outcomes were collected. Then the patency rate and limb salvage rate were calculated.Results:The technique success rate was 71.4% (15/21). The 3, 6, 12 month patency rate were 93.3%, 85.6% and 74.1%, respectively. The 1-year limb salvage rate was 93.3% (14/15). In the 6 patients converted to artificial vessel bypass, the 3,6,12 months patency rates were 76.7%, 66.7% and 46.8%, respectively. The 1-year limb salvage rate was 66.7%.Conclusions:Remote endarterectomy of the lower extremity artery is an alternative option in the treatment of complex ischemic lesions of the lower extremity artery, other than artificial vessel bypass.

Study on Improvement Effects of Polygonum orientale Flower Extract on Hypoxia-reoxygenation Injury of H 9c2 Cardiomyocytes / 中国药房

OBJECTIVE：To study the improvement effects and mechanism of Polygonum orientale flower extract on hypoxia- reoxygenation injury of H 9c2 cardiomyocytes. METHODS ：H9c2 cardiomyocytes were divided into normal control group ，model group and low- ，medium- and high-concentrations groups of P. orientale flower extract （20，40，80 μg/mL）. Except for normal control group ，other groups were given 800 μmol/L CoCl2 to induce hypoxia-reoxygenation injury model. Cell apoptosis was observed. The levels of Ca 2+（in cytoplasm ），mitochondrial membrane potential （MMP），ATP enzyme （Na+-K+-ATP enzyme ，Ca2+-Mg2+-ATP enzyme） activities， the ratio of cytochrome c （Cyto c ）， protein in cytosol to mitochondria ，phosphorylation levels of reperfusion injury salvage kinase （RISK） signaling pathwayrelated protein [protein kinase B （Akt）and extracellular signal regulated kinase 1/2（ERK1/2）] as well as protein expression of HIF- 1 α were detected respectively. In addition，the cells were divided into normal control group ，model group and P. orientale flower extract group （80 μ g/mL），PI3K inhibitor LY294002+CoCl2 group（15 μmol/L LY294002+80 μmol/L ，LY294002+P. orientale flower extract group （15 μmol/L LY294002+80 μg/mL P. orientale flower extract ），MEK inhibitor PD98059+CoCl2 group（25 μmol/L PD98059+800 μmol/L CoCl2），PD98059+P. orientale flower extract group （25 μmol/L PD98059+80 μg/mL P. orientale flower extract ）. After cultured by the same method ，the phosphorylation levels of Akt protein and ERK1/2 protein in the cells were measured to verify the activation of P. orientale flower extract to RISK signaling pathway. RESULTS：Compared with model group ，nuclear pyknosis and the number of apoptotic bodies were reduced in different concentrations groups of P. orientale flower extract. ROS level ，Ca2+ level（except for low-concentration group ），MMP，ratio of Cyto c in cytoplasm to Cyto c in mitochondria ，protein expression of HIF- 1α were decreased significantly（P＜0.05 or P＜0.01）； the activity of ATP enzyme （except for the low-concentration group ），Akt protein and ERK 1/2 protein phosphorylation level were significantly increased （P＜0.01）. After treated with PI 3K inhibitor LY 294002 and MEK inhibitor PD 98059，Akt protein and ERK 1/2 protein phosphorylation level in cadiomyocyte were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：P. orientale flower extract can improve hypoxia-reoxygenation injury of H 9c2 cardiomyocytes，the mechanism of which may be associated with inhibiting cardiomyocyte apoptosis ，improving ATPase activity ，protecting mitochondria ，regulating RISK signaling pathway related proteins and HIF- 1α protein expression.

Study on Anti-inflammatory Effect and Mechanism of Jingulian Capsule on Inflammatory Model Rats / 中国药房

OBJECTIVE：To study the anti- inflammatory effect and mechanism of Jingulian capsule on inflammatory model rats. METHODS ：Totally 48 rats were randomly divided into blank control group ，model group ，Jingulian capsule low-dose ， medium-dose and high-dose groups （0.66，1.32，2.64 g/kg），dexamethasone group （positive control ，0.945 mg/kg），with 8 rats in each group. Blank control group and model group were given constant volume of water intragastrically ，and other groups were given relevant medicine intragastrocally ，twice a day ，for consecutive 3 days. Thirty minutes after last administration ，model group and administration groups were given lipopolysaccharide （10 mg/kg）intraperitoneally to induce inflammatory model. Six hours after intraperitoneal injection ，the contents of TNF-α，IL-1β，IL-6，PGE2 in serum were detected by ELISA. The wet to dry weight （W/D）ratio of lung tissue were determined. HE staining was used to observe the pathological changes of lung tissue . RT-PCR was used to detect the mRNA expression of TNF-α，IL-6，PGE2 and IL- 1β in lung tissue. Western blot assay was used to detect the phosphorylation of NF-κB p65 protein and the expression of IκBα protein in lung tissue. RESULTS：Compared with blank ； control group ，the contents of TNF-α，IL-1β，IL-6 and PGE 2 in serum ，the W/D ratio of lung tissue ，the expression of TNF-α，IL-1β，IL-6 and PGE 2 mRNA and the phosphorylation level of NF-κB p65 protein in lung tissue of model group weresignificantly increased ，and the expression of IκBα proteinwas significantly decreased （P＜0.05 or P＜0.01）；a large number of alveolar atrophy and collapse ，alveolar wall thickening ，lung consolidation ，and a large number of inflammatory cell infiltration could be seen in lung tissue. Compared with model group ，the contents of TNF-α，IL-1β（except for low-dose group ）， IL-6 and PGE 2 in serum ，as well as the expression of TNF-α（except for high-dose group ），IL-1β，IL-6（except for low-dose ， high-dose groups ）and PGE 2 mRNA in lung tissue were decreased significantly in Jingulian capsule groups （P＜0.05 or P＜0.01）； the W/D ratio of lung tissue was decreased significantly in Jingulian high-dose group （P＜0.05 or P＜0.01）；the expression of phosphorylation level of NF-κB p65 protein in lung tissue of Jingulian medium-dose group were decreased significantly （P＜0.05 or P＜0.01），while the expression of IκBα protein was increased significantly（P＜0.05）；the alveolar structure was clear ，the alveolar wall was slightly thickened ， and a small amount of inflammatory cell infiltration was seen in lung tissue. CONCLUSIONS：Jingulian capsule has good anti-inflammatory effect on inflammatory model rats ，the mechanism of which may be related to the inhibition of NF-κB signal pathway.

Development and Evaluation of A CRISPR-based Diagnostic For 2019-novel Coronavirus

BackgroundThe recent outbreak of infections by the 2019 novel coronavirus (2019-nCoV), the third zoonotic CoV has raised great public health concern. The demand for rapid and accurate diagnosis of this novel pathogen brought significant clinical and technological challenges. Currently, metagenomic next-generation sequencing (mNGS) and reverse-transcription PCR (RT-PCR) are the most widely used molecular diagnostics for 2019-nCoV. Methods2019-nCoV infections were confirmed in 52 specimens by mNGS. Genomic information was analyzed and used for the design and development of an isothermal, CRISPR-based diagnostic for the novel virus. The diagnostic performance of CRISPR-nCoV was assessed and also compared across three technology platforms (mNGS, RT-PCR and CRISPR) Results2019-nCoVs sequenced in our study were conserved with the Wuhan strain, and shared certain genetic similarity with SARS-CoV. A high degree of variation in the level of viral RNA was observed in clinical specimens. CRISPR-nCoV demonstrated a near single-copy sensitivity and great clinical sensitivity with a shorter turn-around time than RT-PCR. ConclusionCRISPR-nCoV presents as a promising diagnostic option for the emerging pathogen.

Difference Analysis of Volatile Components in Roots ，Stems and Leaves of Gaultheria yunnane nsis by GC-MS / 中国药房

OBJECTIVE：To compare the volatile components extracted from the roots ，stems and leaves of Gaultheria yunnanensis. METHODS ：Steam distillation method was used to extract the volatile oils from roots ，stems and leaves of G. yunnanensis. Chemical constituents were analyzed by GC-MS. NIST 2011 standard mass spectral library was adopted to select the chromatographic peak with a matching degree higher than 80，and combined with relevant literatures for identification. Relative percentages of chemical constituents were calculated by peak area normalization. RESULTS ：Totally 95 chromatographic peaks were detected in valatile oil from the roots of G. yunnanensis and 54 chemical constituents were identified ，accounting for 82.35％ of the total content of root volatile components. The constituents with relatively high content were methyl salicylate （20.30％）， n-hexadecanoic acid （19.86％），（Z，Z）-9，12-octadecadienoic acid （9.26％），phenanthrene（4.37％）and so on. Totally 69 chromatographic peaks were detected in volatile oil from the stems and leaves of G. yunnanensis and 46 chemical constituents were identified，accounting for 97.10％ of the total content of volatile components in stems and leaves. The constituents with relatively high content were methyl salicylate （86.72％），n-hexadecanoic acid （2.60％），（Z，Z，Z）-9，12，15-octadecatrienoic acid （1.25％） and so on. Both of them contained 16 common constituents such as alkenes esters ，acids and so on. CONCLUSIONS：The chemical constituents of volatile oils extracted from the roots ， stems and leaves of G. yunnanensis are mainly esters and acids. The components are similar to each other ，but the contents of acids in the roots and esters in the leaves and stems are higher.

Study on Metabolism of Miao Medicine Laportea bulbifera Extract in Isolated Human Intestinal Flora / 中国药房

OBJECTIVE：To explore the metabolic charact eristics of Miao medicine Laportea bulbifera extract in isolated human intestinal flora. METHODS ：L. bulbifera was extracted with 70％ ethanol reflux extraction. After concentration，extraction with n-butanol and drying ，L. bulbifera extract was obtained. Taking 0.05 g/mL L. bulbifera extract 1 mL mixed with isolated human intestinal flora fluid 10 mL and cultured for 36 h in anaerobic environment （setting up blank control without drugs or human intestinal bacterial solution ），so as to simulate the metabolic process of the extract in human intestine. The metabolites were detected by UPLC-Q-TOF/MS. The determination was performed on Agilent Eclipse Plus C 18 RRHD column with mobile phase consisted of 0.01％ formic acid water solution- 0.01％ formic acid acetonitrile solution （gradient eluetion ）at the flow rate of 0.25 mL/min. The column temperature was set at 40 ℃，and the sample size was 1 µL. ESI detection was adopted and scanned by negative ion mode （ESI－）；the capillary voltage was 4.5 kV，the ion source temperature was 120 ℃，the collision energy was 15-32 V，and the scanning range was m/z 50-1 000. The “Strip”module of MassLynx V 4.1 software was used to analyze the differential chromatograms between the reaction solution and the blank control of L. bulbifera extract. Mass spectrum data and UNIFI so ftware were used to predict relative molecular weight and formula ；based on the information of substance control and related literature reports ， the structure and biotransformation pathway of L. bulbifera metabolites in isolated human intestinal flora were predicted and analyzed. RESULTS & CONCLUSIONS ： A total of 3 prototype ： products（rutin，quercetin，kaempferol-3-O-rutinoside）and 22metabolites （mainly the metabolites of quercetin ，mono- caffeoylquinic acid ，isoquercitrin，etc.） were detected after metabolized in isolated human intestinal flora. Itsbiotransformation pathway is phase Ⅰ reaction，which mainly consisted of reduction ，oxidation and hydrolysis.

Study on Quality Standard Improvement of Rhizoma Begoniae from Guizhou / 中国药房

OBJECTIVE：To provide reference for improving the quality standard of Rhizoma Bego niae from Guizhou . METHODS：Five batches of Rhizoma Begoniae from Guizhou were collected ，and the microscopic characteri stics of the Rhizoma Begoniae powder were observed. According to the corresponding methods in 2015 edition of Chinese Pharmacopoeia （part Ⅳ）， potent adenosine 50-triphosphate competitive phosphati - dylinositol-3-kinase/mammalian target of rapamycin inhibitors：discovery of compound 26（PKI-587），a highly effi cacious dual inhibitor Morpholine as a privileged structure ：a review on the me - dicinal chemistry and pharmacological activity of morpho - line containing bioactive molecules[J]. Med Res Rev ， qq.com 2019. DOI ：10.1002/med.21634. qualitative identification of Rhizoma Begoniae was conducted by TLC ，and the contents of moisture ，total ash and water-soluble extract in Rhizoma Begoniae were determined. The contents of rutin were determined by HPLC. RESULTS ：The powder of Rhizoma Begoniae medicinal materials was brown ，stone cells were square ，polygonal-like or irregular. There were many starch grains and few complex grains. The conduit ，calcium oxalate square crystal/cluster crystal were visible. The same fluorescence spots were found in the same location of TLC atlas of Rhizoma Begoniae control herb. The moisture ，total ash ，water-soluble extract contents were 10.15％-11.41％，8.70％-12.59％ and 16.91％-19.58％，respectively. The linear range of rutin were 18.47-147.8 μg/mL （r＝0.999 8）；RSDs of reproducibility ，intermediate precision and stability tests （8 h）were all lower than 3.0％；the average recoveries were 99.39％-100.29％（RSDs were 0.23％-2.59％，n＝3）；the contents of rutin in 5 batches of Rhizoma Begoniae were 0.102％-0.198％. CONCLUSIONS ：The contents of moisture and total ash shall not exceed 13.0％ and 14.0％ respectively， and the contents of water-soluble extract and rutin shall not be less than 15.0％ and 0.080％. The quality standard established in this study can be used for the quality control of Rhizoma Begoniae from Guizhou.

Simultaneous Determination of 8 Flavonoid Glycosides in Sedum bulbiferum by HPLC / 中国药房

OBJECTIVE：To establish a metho d for sim ultaneous determination of 8 flavonoid glycosides in Sedum bulbiferum . METHODS：HPLC method was adopted to determine the contents of kaempferol- 3-O-β-D-glucopyranoside-（1→2）-α-L-glucopy- ranoside-7-O-α-L-glucopyranoside（KGGR），kaempferol-3-O-β-D-glucopyranosyl-7-O-α-L-rhamnopyranoside（KGR），quercetin-3- O-α-L-rhamnose-7-O-α-L-rhamnoside（QRR），BulbiferumosideⅡ，kaempferol-3-O-（6-coumarinyl）-β-D-glucose-（1→2）-β-D-glu- cose-7-O-α-L-rhamnoside（KcGGR），kaempferol-3-O-（2-β-D-glucose）-α-L-rhamnose-7-O-α-L-rhamnoside（KGRR），kaempferol-3- O-α-L-rhamnoside-7-O-α-L-rhamnoside（KRR），kaempferol-3-O-（6″-acetyl-β-D-glucose）-7-O-α-L-rhamnoside（KaGR）in S. bulbi- ferum. The determination was performed on Waters CORTECS C 18 column with mobile consisted of acetonitrile - 0.1％ phosphoric acid water solution （gradient elution ）at the flow rate of 0.8 mL/min. The detection wavelength was set at 254 nm，and column temperature was 35 ℃. The sample size was 5 μL. RESULTS：The linear range of 8 constituents were 0.013-0.052，0.005-0.018， 0.008-0.031，0.010-0.042，0.009-0.038，0.008-0.030，0.009-0.037，0.032-0.130 μg，respectively（all r were not less than 0.999 0）. The limits of detection were 0.08，0.14，0.11，0.21，0.42，0.35，0.23，0.28 μg/mL，respectively. The limits of quantification were 0.25，0.47，0.38，0.69，1.40，1.17，0.77，0.93 μg/mL，respectively. RSDs of precision ，reproducibility and stability tests （24 h） were all lower than 3％（n＝6 or n＝7）. The average re coveries were 99.67％-104.20％（RSDs＝0.17％-1.59％，n＝6）. Average contents of above 8 constituents in 13 batches of samples were 0.893 8，0.312 6，0.490 8，0.964 9，0.751 2，0.502 2，0.606 2， 1.915 7 mg/g（n＝3）. CONCLUSIONS ： The method is simple， acourate and reproducible ， and can be used for simultaneous determination of 8 flavonoid glycosides in 才〔2016〕5677） S. bulbiferum .