AIM:To explore the mechanism of ac-tion of the microtubular inhibitor of CA-4 deriva-tive LGD5 on human cervical cancer HeLa cells.METHODS:HeLa cells were selected and divided in-to blank group,CA-4 positive control group,and dif-ferent concentrations of LGD5 were formed into the experimental group.MTT was used to investi-gate the growth inhibition of LGD5 on HeLa cells and to determine the assay concentration.Cell morphology and apoptosis were observed before and after drug administration by inverted micro-scope and acridine orange staining.Immunofluo-rescence staining was used to examine the effect of LGD5 on microtubules using DAPI.The effect of LGD5 on cell cycle by PI flow cytometry.Protein im-munoblotting was used to examine the effect of LGD5 on cyclins and apoptosis-related proteins.RE-SULTS:MTT experiments showed that LGD5 inhibit-ed HeLa cells in a time-and dose-dependent man-ner.Timed photography and acridine orange stain-ing observed that LGD5 induced apoptosis in HeLa cells and produced significant chromatin agglutina-tion and apoptotic bodies.Inhibition of microtu-bule polymerization in HeLa cells by LGD5 was ob-served by DAPI staining.The PI flow cytometry re-sults showed that LGD5 induced G2/M cycle arrest in HeLa cells,was time-dependent and dose-depen-dent within 12 h,and had a significant difference(P＜0.01),apoptosis was induced after 24 h and it was time-dependent.The results of Western blot show that,LGD5 downregulates Cdc 2 and Cdc25C,up-regulation of p-Cdc 2,and Cyclin B1 and p-histone H3,further verified that LGD5 induced G2/M cycle arrest in HeLa cells,besides,LGD5 caused in-creased Caspase 3 expression in HeLa cells,upregu-lated Caspase 9 and Bax,down-regulation of Pro-caspase 9 and Bcl-2,this result indicates that HeLa cell apoptosis induced by LGD5 is related to the mi-tochondrial pathway.CONCLUSION:The CA-4 deriv-ative LGD5 inhibited microtubule polymerization in HeLa cells,induced their G2/M cycle arrest,and subsequently induced cell apoptosis through the mitochondrial pathway.

In this paper, the low-field nuclear magnetic resonance technology CPMG (Carr-Purcell-Meiboom-Gill) echo method was used to determine the cross-linking degree and cross-linking density of crospovidone (PVPP) from different manufacturers. Based on the seven physical properties of PVPP, a fingerprint spectrum (radar chart) of twenty secondary quality indicators were obtained, and three compressibility evaluation indicators, index of the parameter (IP), index of parametric profile (IPP), index of good compression (IGC) were calculated by the fingerprint spectrum. It was found that the cross-linking degree and compressibility index IP of PVPP showed a strong correlation (r = 0.816) by the correlation analysis, indicating that the cross-linking degree is one of the key quality attributes for evaluating the compressibility of PVPP.

Recent studies have showed that thrombosis is closely related to leucocytes involved in immunity. Interfering with the binding of leukocyte integrin Mac-1 and platelet GPIbα can inhibit thrombosis without affecting physiological coagulation. Mac-1-GPIbα is proposed as a potential safety target for antithrombotic agents. Guanxinning tablet (GXNT) is an oral Chinese patent medicine used for the treatment of angina pectoris, which contains phenolic acid active ingredients, such as salvianolic acids, ferulic acid, chlorogenic acid, caffeic acid, rosmarinic acid, tanshinol, and protocatechualdehyde. Our previous studies demonstrated that GXN exhibited significant antithrombotic effects, and clinical studies suggested that it did not increase bleeding risk. In addition, GXN exerted a significantly regulatory effect on immune inflammation. In the current study, we intended to evaluate the effects of GXN on bleeding events and explore the safety antithrombotic mechanism of GXN based on leukocyte-platelet interaction. First, we established a gastric ulcer model induced by acetic acid in rats and found that GXN not only did not increase the degree of gastrointestinal bleeding when gastric ulcer occurred, but also had a certain promoting effect on the healing of gastric ulcer. Second, in vitroexperiments showed that after pretreatment with GXN and activation by phorbol 12-myristate-13-acetate (PMA), the adhesion and aggregation of leukocytes with human platelets were reduced. It was also found that GXN reduced the expression and activation of Mac-1 in leucocytes, and inhibited platelet activation due to leukocyte engagement via Mac-1. Overall, the results suggest that GXN may be a safe antithrombotic agent, and its low bleeding risk mechanism is probably related to inhibited leukocyte-platelet aggregation and its interaction target Mac-1-GPIbα.
Animals ; Fibrinolytic Agents ; Humans ; Integrins ; Leukocytes ; Macrophage-1 Antigen ; Rats ; Stomach Ulcer ; Tablets ; Thrombosis

This study aims to study the effective substance and mechanism of Ziziphi Spinosae Semen extract in the treatment of insomnia based on serum metabolomics and network pharmacology. The rat insomnia model induced by p-chlorophenylalanine(PCPA) was established. After oral administration of Ziziphi Spinosae Semen extract, the general morphological observation, pentobarbital sodium-induced sleep test, and histopathological evaluation were carried out. The potential biomarkers of the extract in the treatment of insomnia were screened by ultra-high performance liquid chromatography-mass spectrometry(UHPLC-MS) combined with multivariate analysis, and the related metabolic pathways were further analyzed. The "component-target-pathway" network was constructed by ultra-high performance liquid chromatography coupled with quadrupole-Exactive mass spectrometry(UHPLC-Q-Exactive-MS/MS) combined with network pharmacology to explore the effective substances and mechanism of Ziziphi Spinosae Semen in the treatment of insomnia. The results of pentobarbital sodium-induced sleep test and histopathological evaluation(hematoxylin and eosin staining) showed that Ziziphi Spinosae Semen extract had good theraputic effect on insomnia. A total of 21 endogenous biomarkers of Ziziphi Spinosae Semen extract in the treatment of insomnia were screened out by serum metabolomics, and the metabolic pathways of phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism, and nicotinate and nicotinamide metabolism were obtained. A total of 34 chemical constituents were identified by UHPLC-Q-Exactive-MS/MS, including 24 flavonoids, 2 triterpenoid saponins, 4 alkaloids, 2 triterpenoid acids, and 2 fatty acids. The network pharmacological analysis showed that Ziziphi Spinosae Semen mainly acted on target proteins such as dopamine D2 receptor(DRD2), 5-hydroxytryptamine receptor 1 A(HTR1 A), and alpha-2 A adrenergic receptor(ADRA2 A) in the treatment of insomnia. It was closely related to neuroactive ligand-receptor interaction, serotonergic synapse, and calcium signaling pathway. Magnoflorine, N-nornuciferine, caaverine, oleic acid, palmitic acid, coclaurine, betulinic acid, and ceanothic acid in Ziziphi Spinosae Semen may be potential effective compounds in the treatment of insomnia. This study revealed that Ziziphi Spinosae Semen extract treated insomnia through multiple metabolic pathways and the overall correction of metabolic disorder profile in a multi-component, multi-target, and multi-channel manner. Briefly, this study lays a foundation for further research on the mechanism of Ziziphi Spinosae Semen in treating insomnia and provides support for the development of innovative Chinese drugs for the treatment of insomnia.
Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal/chemistry* ; Metabolomics ; Network Pharmacology ; Rats ; Seeds/chemistry* ; Sleep Initiation and Maintenance Disorders/drug therapy* ; Tandem Mass Spectrometry ; Ziziphus/chemistry*

The epidemic situation of coronavirus disease 2019(COVID-19) is developing rapidly in the world, and the influence is serious. In this study, the prescription of Mongolian medicine to prevent new type of COVID-19 was investigated. Based on the second edition and the third edition of COVID-19 Mongolian Medicine Prevention and Treatment Guidance Program issued by the Inner Mongolia Autonomous Region Health Commission, using Excel 2007, SPSS Modeler 18, SPSS Statistics 25, Cytoscape 3.7.1 statistical software as a tool, the association rules analysis and cluster analysis of Mongolian medicine included in the standard were carried out. Among the 45 prophylactic prescriptions included in the standard, a total of 34 high-frequency drugs using frequency ≥5 were used, of which Carthami Flos(21 times, 4.46%), Chebulae Fructus(20 times, 4.26%), Moschus(13 times, 2.77%), Myristicae Semen(12 times, 2.55%), Santali Albi Lignum(12 times, 2.55%), and Bovis Calculus(12 times, 2.55%) were the most common. The main drugs for the prevention of COVID-19 were Liang(13 times, 38.23%), Wen(9 times, 26.47%), the flavor was Ku(20 times, 34.48%), Xin(13 times, 22.41%), Gan(11 times, 18.97%), the most used drugs treating hot evil(99 times, 32.46%), treatment of "Heyi" drugs(51 times, 16.72%), treatment of "Badagan" drugs(40 times, 13.11%), treatment of "sticky" drugs(37 times, 12.13%), and a cough, eliminating phlegm and antiasthmatic(31 times, 10.16%), the association rule analysis found that the highest association intensity of the drug pair combination of 11. Clustering analysis using the cluster analysis of inter-group join method found a total of 8 categories. In this study, 45 prescriptions of Mongolian medicine for the prevention of COVID-19 were collec-ted and further analyzed, hoping to provide new ideas for clinical diagnosis and treatment.
Betacoronavirus ; China ; Coronavirus Infections ; drug therapy ; Humans ; Medicine, Mongolian Traditional ; Pandemics ; Pneumonia, Viral ; drug therapy

An analysis method was established by UPLC fingerprint and then applied to simultaneous determination of multiple compounds in Gardeniae Fructus from different areas in China. Samples were separated on a Waters Acquity UPLC BEH C₁₈( 2. 1 mm×50 mm,1. 7 μm) column with 0. 1% formic acid-water and acetonitrile solution as gradient mobile phase at a flow rate of 0. 4 m L·min-1 at various wavelengths. The similarity of samples was over 0. 95 with ″Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine( 2012 edition) ″. The UPLC common fingerprints for 32 batches were established with 19 common peaks identified. The samples were divided into 3 groups analyzed by HCA and PCA. Five components were identified as the main compositions which caused the differences of chemical constituents in the samples from different areas with partial least squares discriminant analysis( PLS-DA). The content of the total components in each area was Zhejiang > Fujian > Jiangxi > Sichuan. This method was accurate and viable,could be used to evaluate the quality of Gardeniae Fructus.
China ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal/standards* ; Fruit/chemistry* ; Gardenia/chemistry* ; Phytochemicals/analysis*

Objective To compare and explore the differences of clinical characteristics between human adenovirus type 7 (HAdV7) and type 55 (HAdV55) infections in adults,and to provide evidences for clinical management.Methods The data of clinical manifestations,laboratory examinations,chest computed tomography and prognosis of 214 cases with HAdV7 and 235 cases with HAdV55 infections from 2012 to 2018 were retrospectively collected and analyzed.The chi-square test was used for the categorical variables,and the rank sum test was used for the continuous variables of non-normal distribution.Results Compared with patients in HAdV55,those in HAdV7 group displayed more diarrhea (12.1％ [26/214] vs 2.6％ [6/235],x2 =15.583),more laryngeal lymphatic follicles (33.2％ [71/214] vs 17.9％ [42/235],x2 =23.566),more tonsil enlargement (56.5％ [121/214] vs 20.0％ [47/235],x2 =63.870) with secretions (33.2％ [71/214] vs 11.5％ [27/235],x2 =30.878),more leukocytosis (24.8％ [53/214] vs 14.0％ [33/235],x2 =8.318),more monocytosis (78.0％ [167/214] vs 52.8％ [124/235],x2 =31.364),more thrombocytosis (7.1％ [15/212] vs 3.8％ [9/235],x2 =5.835),more elevated level of C-reactive protein (80.8％ [97/120] vs 64.3％ [137/213],x2 =10.020),more abnormalities of liver function (alamne aminotransferase [ALT]:13.5％ [28/207] vs 5.6％ [13/232],x2 =8.111) and myocardial enzymes (creatine kinase [CK]:37.6％ [77/205] vs 26.4％ [61/231],x2 =6.246),creatine kinase isoenzymes ([CK-MB]:35.6％ [73/205] vs 11.1％ [24/216],x2 =35.600),and all the differences were statistically significant (all P ＜ 0.05,0.01).Compared with patients in HAdV7 group,those in HAdV55 group displayed longer duration of diseases (7 [5]days vs 6[5] days,Z=-2.632),more sore throat (72.8％ [171/235] vs 62.1％ [133/214],x2 =0.016),more headache (62.1％ [146/235] vs 16.4％ [35/214],x2 =97.527),more pharyngeal congestion (93.6％ [220/235]vs 74.8％ [160/214],x2 =30.602),more leukopenia (14.0％ [33/235] vs 24.8％ [53/214],x2 =8.318),and more elevated level of lactate dehydrogenase (12.6％ [29/230] vs 6.3％ [13/205],x2 =4.881),more pneumonia (71.5％ [168/235] vs31.8％ [68/214],x2 =70.846) (all P＜0.05,0.01).Conclusions The clinical characteristics of HAdV7 and HAdV55 infections in adults are different.The type 55 infection is more likely to develop to pneumonia,while the type 7 infection has wider tissue tropism.

To analyse the quality of three processed products of Rhizome of Curcuma wenyujin by establishing an ultra performance liquid chromatography (UPLC) method for simultaneously determining five sesquiterpene components in three processed products of rhizome of C. wenyujin and establishing UPLC fingerprints. Component determination was achieved on Waters ACQUITY UPLC BEH C₁₈ column（2.1 mm×50 mm, 1.7 μm), with acetonitrile-water as mobile phase for gradient elution. The flow rate was 0.3 mL·min⁻¹; column temperature was 30 °C; the detection wavelength was set at 214 nm and injection volume was 1 μL. The similarity was analyzed with "Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica (2012.130723)", and hydrophobic cluster analysis (HCA), principal components analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were conducted by using simca-p14.1 software to investigate the differences in components among these three kinds of processed products. The curzerene, curdione, curcumol, germacrone, furanodiene and β-elemene showed good linearity relationship with chromatographic peak area within the ranges of 10.8-320（=0.999 9）, 10.36-259（=0.998 1）, 10.54-263.5（=0.999 3）, 30.2-755（=0.999 6）and 34.38-862（=0.999 9）mg·L⁻¹, respectively; their average recoveries were 98.75%, 98.69%, 98.63%, 99.76% and 99.57% respectively, with RSD of 2.67%, 1.47%, 1.29%, 2.54% and 0.87% respectively. The similarity of 30 batches of samples was larger than 0.9, indicating good consistency of the samples. The samples can be clearly classified into three categories for HCA, PCA and OPLS-DA pattern recognition, the differential chromatographic peak among three processed products was found respectively. The results showed that the pharmacology basis had changed obviously after processing of Rhizome of C. wenyujin, so it can provide the scientific basis for rational clinical application and establishing quality standards of three processed products of Rhizome of curcuma wenyujin.

To compare the effects of Curcumae Rhizoma/vinegar-processed Curcumae Rhizomaon immune hepatic fibrosis, proliferation of HSC-T6, and expressions of α-SMA and Procollagen I. The immunological liver fibrosis model was prepared through intraperitoneal injection with porcine serum 0.5 mL in each rat, twice a week, for 14 weeks. Expressions of serum ALT, AST, PC-Ⅲ, IV-C, LN, HA and HYP, MDA in liver tissues were observed after administration of Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma (0.95, 1.90 g•kg⁻¹). The pathological changes in liver tissues were observed by HE staining. Masson staining and Sirius red staining were used to observe the expression of collagen in rat liver. HSC-T6 was cultured, and the proliferation of HSC-T6 was determined by MTT assay at different concentrations in 12, 24, 36, 48 h. The expressions of α-SMA and Procollagen I were detected by Real-time PCR. The results showed that expressions of serum ALT, AST, PC-Ⅲ, IV-C, LN and HA in Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma groups (0.95, 1.90 g•kg⁻¹) were significantly lower than model group; in terms of effect, vinegar-processed Curcumae Rhizoma group was superior to Curcumae Rhizoma group. Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma containing serum could inhibit the proliferation of HSC-T6 in a dose-effect and time-effect manner. Expressions of α-SMA and Procollagen I in HSC-T6 were decreased after 24 h, especially in 20% vinegar-processed Curcumae Rhizoma containing serum group (P<0.01). Both Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma could reduce immune hepatic fibrosis to varying extent. Their anti-hepatic fibrosis mechanism may be correlated with inhibition of the proliferation of HSC-T6, and reduction of the formation of extracellular matrix and promotion of its degradation.

The essay summarizes the current development and application of reference extracts from traditional Chinese medicines (TCMs). Through the analysis on problems in the quality control of TCMs, and on the basis of the current application of reference extracts in Chinese Pharmacopoeia, it investigates reference extracts from TCMs and forecasts the study orientation on TCM quality standards. Despite a wide prospect for the application of reference extracts in quality control of TCMs, there remain problems in varieties and standards. It is of great significance for the TCM understanding to further promote the development of reference extracts.
China ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Medicine, Chinese Traditional ; methods ; standards ; Pharmacopoeias as Topic ; standards ; Quality Control ; Technology, Pharmaceutical ; methods ; standards