To observe the effects of early intervention with effective components from a Chinese herbal formula (Huannao Yicong formula, HNYCF) on behavior and related indicators of cholinergic system in β-amyloid precursor protein (APP) transgenic mice.

Objective To study the effects of imatinib mesylate on the apoptosis in human melanoma cell line M14.Methods M14 cells were cultured in vitro in the presence of imatinib mesylate at three concentrations(5,10 and 20 μmol/L)for 96 hours.Sebsequeutly,annexin V-FITC and propidium iodide(PI)double staining flow cytometry and terminal deoxvnucleotidvl transferase (TdT)-mediated dUTP nick end labeling (TUNEL)were used to detect the cell cycle and apoptosis,respectively,DAPI staining to observe the mot-phological changes.and Western blot to measure the protein expressions of bcl-2 and bax in cells.Results Imatinib mesylate of the three concentrations could induce an evident increase in the apoptosis in M14 cells.Compared with untreated M14 cells,an increase of cell population in S phase was observed in imatinib mesylate.treated cells(P<0.05),along with a decline in cell population in G2/M phases(P<0.01).Annexin V/PI double staining and TUNEL revealed a significant increase in the rate of early apoptosis and in the acount of apoptotic cells,respectively,in M14 cells treated with imatinib mesylate of the three concentrations(all P<0.01).After treated with imatinib mesylate of 20 μmol/L.there was a morphological change characteristic of apoptosis in M14 cells,together with an upregulated expression of bcl-2(t=15.46,P<0.01)and downregu-lated expression of bax(t=25.53,P<0.01).Conclusions Imatinib mesylate can interfere with the process of cell cycle of and induce the apoptosis in M14 cells,which may be mediated through mitochondrial pathway.

A new hasubanan alkaloid, hernsubanine E (1), as well as two known compounds p-hydroxybenzaldehyde (2) and (-)-syringaresinol (3) have been isolated from the whole plants of Stephania hernandifolia by various column chromatographic methods. Their structures were identified by physicochemical properties and spectral analyses. Compounds 2 and 3 were isolated from the genus of Stephania for the first time.
Alkaloids ; chemistry ; Heterocyclic Compounds, 4 or More Rings ; chemistry ; isolation & purification ; Stephania ; chemistry

OBJECTIVE To clarify out the network pharmacology mechanism of Polygala tenuifolia against Alzheimer disease(AD).METHODS Firstly,we collected the chemical constituents from Polyg-ala tenuifolia and key targets toward AD.Machine learning algorithms were applied to construct classifi-ers for predicting the effective constituents. Secondly, docking models were utilized for further evalua-tion.Finally,we built constituent-target,target-target network and target-biology pathway network.RE-SULTS 104 chemical constituents Polygala tenuifolia from were collected.Through prediction of blood-brain penetration and validation,36 chemical constituents were selected among 100 chemical constitu-ents,their action targets mainly focused on AChE,COX-2,TNF-α,insulin-degrading enzyme and APP. Their main structure types include Polygala saponins, Polygala glycosides, Polygala shrubby ketones, polygala xanthones and sterols,which acted on AchE,APP,M-TAU,GSK3β and 5HT1A with high fre-quency.Gene-Ontology and KEGG enrichment analysis showed that the main pathways of these con-stituents involve in neurotransmitter release,synaptic conduction and synaptic plasticity,apoptosis reg-ulation,phosphorylation pathway,Ca2+signaling pathway,and so on.CONCLUSION This study uncov-ered a network mechanism of Polygala tenuifolia against Alzheimer disease,which may provide impor-tant information for the further study and new drug development.

Influenza caused by influenza virus,seriously threaten human life and health.Drug treatment is one of the effective measurement. However, there are only two classes of drugs, one class is M2 blockers and another is neuraminidase (NA)inhibitors. The recent antiviral surveillance studies reported a global significant increase in M2 blocker resistance among influenza viruses, and the resistant virus strains against NA inhibitor are also reported in clinical treatment.Therefore thediscovery of new medicines with low resistance has become very urgent.As all known,traditional medicines with multi-target features and network mechanism often possess low resistance. Compound Yizhihao, which consists of radix isatidis,folium isatidis,Artemisia rupestris,is one of the famous traditional medicine for influenza treatment in China, however its mechanism of action against influenza is unclear. In this study, the multiple targets related with influenza disease and the known chemical constituents from Compound Yizhihao were collected, and multi-target QSAR (mt-QSAR) classification models were developed by Na?ve Bayesian algorithm and verified by various datasets. Then the classification models were applied to predict the effective constituents and their drug targets.Finally,the constituent-target-pathway network was constructed,which revealed the effective constituents and their network mechanism in Compound Yizhihao. This study will lay important basis for the clinical uses for influenza treatment and for the further research and development of the effective constituents.

Objective To prepare quercetin ( QT )-loaded polylactic-co-glycolic acid-D-α-tocopheryl polyethylene glycol 1000 succinate ( PLGA-TPGS) nanoparticles ( QPTN) and QT-loaded polylactic-co-glycolic acid ( PLGA) nanoparticles ( QPN) by using QT as model drug and PLGA-TPGS or PLGA as carrier materials, and to investigate the quality of the two nanoparticles. Methods QPTN and QPN were prepared by using the ultrasonic emulsification-solvent evaporation method, and their surface morphology,size and surface charge were detected by using a transmission electron microscope ( TEM) and a Nano ZS90 light scattering and laser Doppler anemometry, respectively. Drug loading ( DL) , entrapment efficiency ( EE) and in vitro drug release of QT in the two nanoparticles were determined by using a reverse phase-high performance liquid chromatography (RP-HPLC) on Hypersil C18 column (4.6 mm×250 mm, 5 μm) with methanol and 0.03% phosphoric acid (3︰2) as mobile phase, and the detective wavelength was 370 nm. Results TEM images exhibited that two nanoparticles were all spherical and regular. The average sizes of QPTN and QPN were (155.4±2.7) nm and (363.8±3.2) nm, while DL and EE of QPTN were approximately (21.6±2.8)%, (93.7±2.9)% (n=6), and DL and EE of QPN were approximately (15.0±1.5)%, (64.6± 1.6)% (n=6), respectively. Both of nanoparticles exhibited sustained release, and the cumulative QT release of QPTN and QPN reached (85.8±2.8)% and (68.6±1.4)% (n=6) at day 30, respectively, with a significant difference between them (P<0.05) . Conclusion QPTN gets smaller size, higher DL and EE, and exhibits sustained release, and the in vitro cumulative QT release is faster and more complete than QPN relatively.

OBJECTIVETo study the effects of Huannao Yicong formula (HNYCF) extract on behavior and ultrastructure of mitochondria in hippocampus CA1 area of APP transgenic mice of different months, and explore its partial mechanism in treating Alzheimer's disease (AD) through the perspective of energy metabolism.

METHODOne hundred and twenty APP695V717I transgenic mice of 3-month old were divided randomly into model group, Donepezil group (0.65 x 10(-3) g x kg(-1) x d(-1)), HNYCF extract large dose group (2.8 g x kg(-1) x d(-1)) and HNYCF extract small dose group (1.4 g x kg(-1) x d(-1)), and 30 mice in each group. Another 30 C57BL/6J mice with the same age and background were used as normal control group. All animals were administered once daily by gavage with the corresponding drug or distilled water. The course of intervention was 4 and 6 months. Behavioral changes were observed by Morris water maze test and step down test. Ultrastructure of mitochondria in hippocampus CA1 area was observed by transmission electron microscope.

RESULTAt the age of 7 and 9 month, the number of times of passing through platform, swimming time and path length of model group increased significantly (P < 0.05, P < 0.01) in Morris water maze test, and the latent period decreased (P < 0.01) in step down test compared with normal group, and it would get worse with the development of disease course. HNYCF extract could increase the number of times of passing through platform, swimming time and path length (P < 0.05, P < 0.01) in Morris water maze test, prolong latent period in step down test of different age. At the age of 7 and 9 month, mitochondrial of hippocampus CA1 area was disrupted and dissolved. Most ridge structure arranged in a mess, and some ridge showed expanding, matrix loosing and swollen appearance, and it would get worse with the development of disease course. HNYCF extract could improve ultrastructure of mitochondria in hippocampus CA1 area, and increase its quality.

CONCLUSIONLearning and memory ability decreased in APP transgenic mice model, and the quantity of neural mitochondria in hippocampus CA1 area with structure disrupting, and it would get worse with the development of disease course. HNYCF extract could improve the learning and memory ability of APP transgenic mice model, its mechanism might relate with improving ultrastructure of mitochondria in hippocampus, and increasing its quantity.

Age Factors ; Alzheimer Disease ; drug therapy ; pathology ; physiopathology ; Animals ; Behavior, Animal ; drug effects ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Female ; Hippocampus ; drug effects ; ultrastructure ; Male ; Maze Learning ; drug effects ; Medicine, Chinese Traditional ; Memory ; drug effects ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Mitochondria ; drug effects ; ultrastructure ; Random Allocation ; Specific Pathogen-Free Organisms

Objective:To explore the clinical efficacy of percutaneous vertebroplasty(PVP) in the treatment of osteoporotic vertebral compression fractures(OVCF) patients with chronic kidney disease-mineral and bone disorder(CKD-MBD), and analyzed the efficacy of the operation in relieving pain and improving quality of life.Methods:This retrospective study selected 71 patients who underwent PVP treatment for OVCF at Beijing Friendship Hospital, Capital Medical University from December 2013 to December 2018. Among them, there were 24 males and 47 females, with an age range of 66-92 years and an average age of (73.7±8.4) years. Based on whether the patients had CKD-MBD, the patients were divided into two groups: 31 patients with CKD-MBD comprised the experimental group, and 40 patients without CKD-MBD comprised the control group. General patient information and perioperative data were collected, including surgical time, bone cement fill volume, preoperative, postoperative, and different follow-up timepoint visual analog scale (VAS) pain scores, analgesic medication usage scores, oswestry disability index (ODI) scores. Measure and record patient vertebral anterior, middle, and posterior heights and Cobb′s angle, as well as patient blood calcium, blood phosphorus, bone metabolic markers, serum 25-hydroxyvitamin D, parathyroid hormone, total hip bone density, femoral neck bone density, and bone fracture indicators. Measurement data were represented as mean±standard deviation( ± s), the comparison between groups was conducted using the t-test; and repeated measure ANOVA was used for comparison before and after operation; the comparison of count data between groups was conducted by Chi-square test. Results:The surgical duration for the patients in this group was 20-50 min, average (29.8±7.2) min, and the volume of bone cement used was 2.0-5.0 mL, average (3.0±1.0) mL. In the experimental group, VAS scores of postoperative pain, analgesic medication usage scores, and ODI showed statistically significant differences compared to preoperative values ( P<0.001). At the last follow-up, there were no statistically significant differences in analgesic medication usage scores and ODI compared to postoperative values, but VAS scores had improved to a certain extent compared to postoperative values, with statistical significance ( P<0.001). In the experimental group, vertebral anterior height increased from (2.26±0.20) cm preoperatively to (2.57±0.28) cm postoperatively, and vertebral middle height increased from (1.96±0.18) cm preoperatively to (2.21±0.16) cm postoperatively, both with statistically significant differences ( P<0.001). Three patients (9.7%) experienced recurrent fractures, including 1 case of surgical vertebral recurrent fracture (3.2%). The experimental group showed a general increasing trend in blood calcium levels, with the last follow-up blood calcium being (2.31±0.09) mmol/L, which was significantly higher than preoperative ( P=0.002). There was no statistically significant difference in the changes in blood phosphorus ( P>0.05), and parathyroid hormone levels showed a slight decrease in the last follow-up when compared to preoperative, but the difference was not statistically significant ( P>0.05). Both total hip bone mineral density(BMD) and femoral neck BMD at the last follow-up showed significant increases compared to preoperative values. The experimental group had higher levels of blood phosphorus and parathyroid hormone than the control group at both preoperative and last follow-up assessments, with statistical significance ( P<0.05). Conclusion:PVP can effectively alleviate pain and enhance the quality of life for patients with OVCF accompanied by CKD-MBD.

BACKGROUND: Arteriosclerosis obliterans (ASO) is a major cause of adult limb loss worldwide. Autophagy of vascular endothelial cell (VEC) contributes to the ASO progression. However, the molecular mechanism that controls VEC autophagy remains unclear. In this study, we aimed to explore the role of the GRB2 associated binding protein 1 (GAB1) in regulating VEC autophagy. METHODS: In vivo and in vitro studies were applied to determine the loss of adapt protein GAB1 in association with ASO progression. Histological GAB1 expression was measured in sclerotic vascular intima and normal vascular intima. Gain- and loss-of-function of GAB1 were applied in VEC to determine the effect and potential downstream signaling of GAB1. RESULTS: The autophagy repressor p62 was significantly downregulated in ASO intima as compared to that in healthy donor (0.80 vs. 0.20, t = 6.43, P < 0.05). The expression level of GAB1 mRNA (1.00 vs. 0.24, t = 7.41, P < 0.05) and protein (0.72 vs. 0.21, t = 5.97, P < 0.05) was significantly decreased in ASO group as compared with the control group. Loss of GAB1 led to a remarkable decrease in LC3II (1.19 vs. 0.68, t = 5.99, P < 0.05), whereas overexpression of GAB1 significantly led to a decrease in LC3II level (0.41 vs. 0.93, t = 7.12, P < 0.05). Phosphorylation levels of JNK and p38 were significantly associated with gain- and loss-of-function of GAB1 protein. CONCLUSION Loss of GAB1 promotes VEC autophagy which is associated with ASO. GAB1 and its downstream signaling might be potential therapeutic targets for ASO treatment.
Adaptor Proteins, Signal Transducing ; Adult ; Arteriosclerosis Obliterans/genetics* ; Autophagy ; GRB2 Adaptor Protein ; Humans ; Phosphoproteins/metabolism* ; Phosphorylation ; Protein Binding ; Signal Transduction

OBJECTIVETo study the effects of Huannao Yicong Recipe (HNYCR)extract on the learning and memory ability, and the expressions of amyloid precursor protein (APP), beta-site APP-cleaving enzyme 1 (BACE1), presenilin-1 (PS-1), and beta amyloid protein (Abeta)in hippocampus CA1 area of APP transgenic mice, and to explore its mechanisms for treating Alzheimer's disease (AD).

METHODSTotally 3-month-old APP695V7171 transgenic mice were used to establish the AD model in this research. They were randomly divided into the model group, the Donepezil group, the large dose HNYCR extract group, the small dose HNYCR extract group, and the normal control group (C57BL/6J mice), 15 in each group. These animals were gavaged for 4 continuous months. Relevant indicators were detected: Morris water maze test was used to measure the spatial learning and memory ability. The immunohistochemical assay was used to detect the expressions of APP, BACE1, PS-1, and Abeta.

RESULTSThe times of crossing the original platform and the swimming time and distance in the fourth quadrant of the 7-month-old APP transgenic mice were significantly reduced in Morris water maze test, when compared with the normal control group (P < 0.01). The times of crossing original platform and the swimming time and distance in the fourth quadrant of all treatment groups significantly increased in Morris water maze test, when compared with the model group (P < 0.05). The expressions of APP, BACE1, PS-1, and Abeta in hippocampus CA1 area of 7-month-old model mice increased significantly (P < 0.01), when compared with the normal control group. The expressions of APP, BACE1, PS-1, and Abeta in each 7-month-old intervention groups were significantly reduced, when compared with the model group (P < 0.01).

CONCLUSIONEarly application of HNYCR extract can obviously improve the learning and memory ability of APP transgenic mice that has declined, reduce the expressions of APP, BACE1, PS-1, and Abeta in the hippocampal CA1 area, reduce the production of Abeta, and slow down the pathological process of brains in APP transgenic mice.

Alzheimer Disease ; metabolism ; Amyloid Precursor Protein Secretases ; genetics ; metabolism ; Amyloid beta-Peptides ; genetics ; metabolism ; Amyloid beta-Protein Precursor ; genetics ; metabolism ; Animals ; Aspartic Acid Endopeptidases ; genetics ; metabolism ; Brain ; drug effects ; metabolism ; CA1 Region, Hippocampal ; drug effects ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Female ; Male ; Maze Learning ; drug effects ; Memory ; drug effects ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Presenilin-1 ; genetics ; metabolism