The changes of bioactive constituents were analyzed for Puhuang-Wulingzhi before and after compatibility and the antiplatelet and antithrombin activitives were evaluated in order to elucidate the scientific and reasonable of Puhuang-Wulingzhi compatibility. UPLC-QTOF-MA-Markerlynx, principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis were used for data analysis and tracking changes of chemical composition during the decocting process. In vitro platelet aggregation induced by ADP, thrombin time(TT) and prothrombin time (PT) were investigated for Puhuang-Wulingzhi before and after compatibility. The results showed that significant differences were found between the mixed decoction and codecoction of Wulingzhi and Puhuang. Five compounds changed obviously were identified as typhaneoside, naringenin, isorhamnetin-3-O-ruinoside, quercetin-3-O-neohesperidoside, kaempferol-3-O-neohesperidoside. The codecoction, comparing with the single decoction, was more significant in antiplatelet aggregation and could prolong thrombin time. In the same crude drug dose, the thrombin time (TT) elongation were greater. These data could provide references for elucidation of bioactive components for this herb pair.
Animals ; Antithrombins ; chemistry ; pharmacology ; Blood Platelets ; drug effects ; physiology ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Humans ; Molecular Structure ; Platelet Aggregation ; drug effects ; Rabbits ; Thrombin Time

Objective To analyze the expression profile of circular RNA (circRNA) in LPS-treated murine peritoneal macrophages (MPMs) and to investigate the effects of mmu_circ_0000790 (circ790) on the secretion of pro-inflammatory cytokines in MPMs following LPS stimulation.Methods MPMs were isolated from C57BL/6 male mice and then stimulated with (LPS treatment group) or without (blank control group) 1 mg/L of LPS for 12 hours.Supernatants of cell culture and cell pellets were collected from each group.Enzyme linked immunosorbent assay (ELISA) was used to measure the changes in the secretion of IL-1β,IL-6 and TNF-α in the supernatants.Expression profile of circRNA in macrophages from the two groups (n=3) was analyze by circRNA microarray.Some differentially expressed circRNAs were validated by real-time PCR (RT-PCR).Moreover,RT-PCR was also performed to detect the expression of circ790 in MPMs after LPS stimulation.Small interfering RNA (siRNA) oligo specific for circ790 was designed and synthesized.Then the synthesized siRNA oligo and normal control (NC) oligo were transfected into MPMs by LipofectamineTM2000.The transfected MPMs were treated with LPS.ELISA was used to detect the levels of IL-1β,IL-6 and TNF-α in the supernatants.Arraystar's home-made microRNA target prediction software was used to predict circ790/microRNA.Results The concentrations of IL-1β,IL-6 and TNF-α in the LPS treatment group were significantly higher than those in the blank control group (P<0.01).These results indicated that the inflammatory model of MPMs was successfully constructed.Statistical differences in 34 differentially expressed circRNAs were found between the two groups (P<0.05).Among them,12 circRNAs were up-regulated and the other 22 circRNAs were down-regulated in the LPS treatment group.RT-PCR results were generally consistent with the microarray data.The expression of circ790 was increased by (1.94±0.15),(3.18±0.13) and (4.21±0.22) folds as compared with that of the blank control group at 3,6 and 12 h after LPS stimulation (P<0.05).After transfecting the MPMs with circ790-siRNA,the secretion of IL-6 was significantly decreased (P<0.05) without notable influence on the secretion of IL-1β and TNF-α.circ790 could function as a microRNA sponge to regulate the gene expression.Conclusion These data show a significantly altered circRNA expression profile in the LPS-treated MPMs.circ790 may be involved in the regulation of IL-6 secreted by macrophages.

0.05).Overall prevalence of UI was 15.93% in the students,20.45% in those of junior high school,10.44% in senior high school and 16.72% in university(P0.05),accounting for 63.90%,10.47% and 25.63% of the total students with UI.Conclusions Great importance should be attached to the higher prevalence of LUTS in young and adolescent females by gynecologists and urologists.More attention should also be paid to health education on LUTS and medical care for those with LUTS to alleviate and delay occurrence of UI symptoms.

Objective To explore the efficacy of Compound Qianliean Granule in the treatment of chronic nonbacterial prostatitis study in rats. Methods The rats were randomly divided into three groups, experimental group ( treated with 3.0 g? kg -1 Compound Qianliean Granule ) and control group ( treated with 2.4 g? kg-1 Qianlieshule granule ) and normal group ( treated with distilled water ) , 1 times daily. The difference of white blood cell count, lecithin corpuscle density were observed after 7 days administration by microscope.The difference of left and right ear weight was calculated on the degree of swelling. The differences of quantity in abdominal cavity dye of relatively deep was compared.Results The white blood cell count in experimental group was significantly higher than that of the control group and the normal group.The lecithin corpuscle density as compared with the control group and the normal group, the difference is statistically significant (P<0.05).The difference of swelling degree and the absorbance compared between experimental group, control group and normal group were significant ( P<0.05).Conclusion Compound Qianliean Granule has good inhibitory effect on chronic abacterial prostatitis.

Objective To study the effects of lidocaine and dexamethasone treatment on tissue damage caused by dopamine extravasation in children.Methods Totals of 47 cases with tissue damage caused by dopamine extravasation were randomly divided into observation group ( 24 cases ) and conventional group (23cases).The observation group received the treatment of 2％ lidocaine1ml,dexamethasone 1 mg and saline1ml to partially closure,while the conventional group received the treatment of 50％ magnesium sulfate to partially wet.Then,therapeutic effects of two groups were observed.Results The therapeutic effects of observation group which 17cases were totally cured and 6cases were effectively cured better than that of conventional group which 5cases were totally cured and 8cases were effectively cured,and the difference was statistically significant ( x2 =4.1155,P ＜ 0.05 ).Conelusious The treatment of lidocaine and dexamethasone have an significantly effects on children with tissue injury coursed by dopamine extravasation.

In order to evaluate the effect and mechanism of the mulberry leaf alkaloid, flavones, and polysaccharide intervention on diabetes, the overall metabolite profiling characteristics for the plasma of diabetic mouse was performed by using an ultra-performance liquid chromatography/electrospray-tandem mass spectrometry (UPLC-ESI-MS). The 8 potential biomarkers were found in diabetic mice plasma based on the data of MS/MS characteristics obtained from the UPLC-OrbitrapMS analysis, which mainly involved in sphingolipids, amino acid metabolic pathway. The principal component analysis showed that the normal group and model group were obviously distinguished and implied that metabolic disturbance was happened in diabetic mice plasma. The extracts of mulberry leaf flavonoids, polysaccharide, alkaloid had exhibited the effects of callback function for diabetic mice through regulating the amino acid metabolism and sphingolipid metabolism.
Alkaloids ; chemistry ; Amino Acids ; metabolism ; Animals ; Biomarkers ; blood ; Chromatography, High Pressure Liquid ; Diabetes Mellitus, Experimental ; drug therapy ; Flavones ; chemistry ; Flavonoids ; chemistry ; Metabolic Networks and Pathways ; Metabolomics ; Mice ; Morus ; chemistry ; Plant Leaves ; chemistry ; Sphingolipids ; metabolism ; Tandem Mass Spectrometry

To investigate the optimum harvesting time and utilization of mulberry leaves during different growth periods based on the content of alkaloids and flavonoids, 88 samples of 11 species of mulberry leaves were collected and analyzed. UPLC-TQ/MS method was applied and the results showed that the ingredients of alkaloids and flavonoids in mulberry leaves are quite different in different growth periods and different species. There was a sharp decline of the average content of alkaloids in all samples from October, while the content of flavonoids dropped either from October but with less volatile. The content of flavonoids in M. atropurpurea was much higher than alkaloids, while M. australis was opposite completely. There was a sharp decline of alkaloids in M. cathayana and M. mongolica from Tuly to August, however, the content of alkaloids and flavonoids in M. alba is neither too high nor too low. In summary, it is more suitable to harvest tender mulberry leaves harvested from the end of September to beginning of October that provide a scientific evidence for rational harvest and comprehensive utilization of mulberry leaves.
Alkaloids ; analysis ; isolation & purification ; Chromatography, High Pressure Liquid ; Drug Stability ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Flavonoids ; analysis ; isolation & purification ; Mass Spectrometry ; Medicine, Chinese Traditional ; Morus ; chemistry ; Plant Leaves ; chemistry ; Plants, Medicinal ; Reproducibility of Results ; Seasons

The present study is to determine the flavonoid glycosides, terpene lactones, biflavones, gingko acid and procyanidins of ginkgo pollen. UPLC-TQ-MS technology was used for the determination of 24 kinds of resource chemical composition in ginkgo pollen qualitatively and quantitatively. The results shows that the contents of rutin, quercetion 3-O-[4-O-(α-L-rhamnosyl )-β-D-glucoside] and kaempferolis were 120.9, 114.0, 222.1 μg x g(-1). In this paper, the contents of 24 kinds of chemical components of ginkgo pollen were determinated by UPLC-TQ-MS for the first time. This method is simple and quick, which will be benefit for recycling utilization of ginkgo pollen.
Chromatography, High Pressure Liquid ; methods ; Flavonoids ; analysis ; Ginkgo biloba ; chemistry ; Mass Spectrometry ; Pollen ; chemistry ; Proanthocyanidins ; analysis ; Rutin ; analysis ; Terpenes ; analysis

To study the fingerprint of the volatile oil of Atractylodes lancea (Thunb.) DC., and to offer the characteristic data for the quality evaluation, GC-MS analysis was performed for 17 samples of different areas used as Atractylodes lancea (Thunb.) DC. Nine kinds of same components were selected. TIC profiles were evaluated by "Computer Aided Similarity Calculation". The characteristic peaks in chromatograms were identified by comparing mass data with literatures. Hierarchical clustering analysis was performed by SPSS based on the relative peak area (RPA) of identified peak to atractydin in 17 samples. The mutual mode fingerprint plots of genuine Atractylodes lancea (Thunb.) DC. have been established, the matching of active components was characteristic that atractylon, hinesol, beta-eudesmol, atractydin as (0.89 - 1.12): (0.11 - 0.15) : (0.48 - 0.61) : 1. The difference of resemblance of not genuine samples with genuine samples was remarkable. Two categorizations were clustered. Atractylodes lancea (Thunb.) DC. from genuine and Tangshan and Nanshan, Jiangsu Prov. were in a group, while those from Anhui and Hubei Prov. were in another group. The characteristic fingerprint of genuine Atractylodes lancea (Thunb.) DC. combined with the study of the matching of active components for the quality control and the resemblance calculation of fingerprints and SPSS hierarchical clustering analysis provided a new analytic method for the quality evaluation and discrimination of Atractylodes lancea (Thunb.) DC.
Atractylodes ; chemistry ; Cluster Analysis ; Ecosystem ; Gas Chromatography-Mass Spectrometry ; Oils, Volatile ; chemistry ; isolation & purification ; Plant Oils ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Quality Control ; Rhizome ; chemistry ; Sesquiterpenes ; analysis ; Sesquiterpenes, Eudesmane ; analysis ; Spiro Compounds ; analysis

To generate transgenic mice in which both hygromycin (hyg) and neomycin (neo) resistance genes are expressed in murine fibroblast cells (MEFs), which are required for conditional gene knock-out and screening of drug resistant ES cell clones. To construct HygR-neoR expression vector, pTK-hygR-pA and PGK-neoR-pA were cloned into pBluescript vector. DNA fragments of tandem genes ( 4245bp ) were prepared by Kpn I and Xba I digestion and transgene was microinjected into pronucleus of zygotes to generate transgenic mice. Transgenic mice were identified by PCR and Southern blot; expression of hygR and neoR gene transcripts were detected by RT-PCR. 7 founder mice carrying hyg-neo resistant genes were obtained and 6 transgenic mouse lines were successfully established. The hygR and neoR gene transcripts were detected in the liver and/or ovary of transgenic mice from hn30, hn33, hn66 and hn67 mouse lines. In MEFs isolated from the mice of line hn66 and hn30, expression of hyg and neo resistant genes was also detectable. Transgenic mouse lines expressing two anti-drug genes have been established. The hyg and neo resistant gene transcripts were detected in the MEFs of two transgenic mouse lines.
Animals ; Cinnamates ; pharmacology ; Drug Resistance, Multiple ; genetics ; Fibroblasts ; metabolism ; Hygromycin B ; analogs & derivatives ; pharmacology ; Mice ; Mice, Transgenic ; Neomycin ; pharmacology ; Transgenes ; genetics