In order to affirm the cardioactive components in Fuzi, we identified a group of aminoalcohol- diterpenoid alkaloids in Fuzi using ultra high-performance liquid chromatography coupled with electrospray ionization mass spectrometer (UPLC-ESI-MS) method. Among a total of forty-one isolated ingredients, thirteen major aminoalcohol-diterpenoid alkaloids were identified by comparing their retention times and MS spectra with those of the reference substances. Moreover, Fuzi samples from different places of origin and with different processing methods were examined and their components displayed a pattern of high similarity, though the relative abundance varies probably due to their different processing methods. Furthermore, the cardiac effect of each identified alkaloid was individually evaluated using the isolated bullfrog heart perfusion experiment. Among the thirteen aminoalcohol diterpenoid alkaloids tested, six of them significantly enhanced the amplitude rates. Taken together, we affirm that the cardioactive components in Fuzi are aminoalcohol-diterpenoid alkaloids, shedding light on future studies of the mechanisms and development of these cardioactive compounds.
Aconitum ; chemistry ; Alkaloids ; chemistry ; Amino Alcohols ; chemistry ; Animals ; Cardiotonic Agents ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Heart ; drug effects ; In Vitro Techniques ; Plant Extracts ; chemistry ; Rana catesbeiana ; Spectrometry, Mass, Electrospray Ionization

Antioxidants ; metabolism ; pharmacology ; Down-Regulation ; Gene Expression Regulation ; Humans ; NF-E2-Related Factor 2 ; genetics ; metabolism ; physiology ; Neoplasms ; genetics ; metabolism ; Oxidative Stress ; genetics ; physiology ; Response Elements ; physiology ; Signal Transduction ; physiology

Objective To investigate the discrepancy of aldosterone synthesis process and potential regulation abnormality between aldosterone-producing adenoma(APA) and normal adrenal(NA) with microarray. Methods cRNA probes labelled with biotin were prepared from mRNA of APAs(APA group,n=10) or NAs(control group,n=7).The probes were hybridized with oligonucleotide microarray of target gene expression profile.Expression levels were read from the fluorescent intensity scanned.The difference of gene expression profile was analyzed by computer software.Differentially expressed genes were verified by real-time RT-PCR. Results Compared with control group,97 genes were up-regulated and 168 genes were down-regulated in APA group.In the genes related to steroid hormone synthesis,only CYP11B2 was significantly up-regulated.In the physiologic regulators of aldosterone synthesis,CYB5A,CYP17A1,DUSP1 and HMGCR were down-regulated,while RENBP and NR1H2 were up-regulated.As a key enzyme in the biosynthesis of cortisol,the expression of CYP17A1 gene was inhibited. Conclusion Among the aldosterone synthesis related enzyme and corresponding regulatory genes in APA,CYP11B2 may be a key synthetase,and the suppressed physiologic regulators of aldosterone synthesis may indicate the existence of neoplastic modulation.

Objective To study the discharges regularity of ambulatory electroencephalogram (ambulatory,EEG,AEEG)during sleep and hyperventilation(HV)-induced EEG. Methods Features of epileptiform discharges of AEEG and HV-induced EEG were evaluated comparatively in 65 cases with frontal lobe epilepsy. Results The epileptiform discharge rate of HV-induced EEG was evidently lower than that of AEEG during the shallow sleep period (non-rapid-eye-movement phase 1 and 2,NREM phase 1 and 2),which had statistical significance(P＜0.01);however,the rate of HV-induced EEG had no significant difference from that of AEEG during the awake period and deep sleep period(NREM phase 3 and 4)(P＞0.05). Conclusions The epileptiform discharge rate of AEEG during the shallow sleep period is obviously higher than that of HV-induced EEG in patients with frontal lobe epilepsy,and thus sleep EEG is helpful to enhance the diagnostic rate of epileptiform discharges in these patients.

OBJECTIVETo assess the risk factors for vulvar dystrophy.

METHODSAn epidemiological study was carried out. Data on 100 cases with vulvar dystrophy was reviewed and face to face interviewed with a uniform questionnaire including the manner of work, environmental temperature, habit of eating, mood, underwear wearing, autoimmune diseases, marriage, menstrual age, the quantity of menses, orders of pregnancy, and labor trauma of vulvar during delivery, vulvitis and urethritis ect. Univariable analysis and multivariate logistic regression analysis were carried out with 1:1 case-control methodology.

RESULTSMultiple conditional logistic regression analysis showed that vulvar dystrophy was positively associated with hot food (OR = 2.55, 95% CI: 1.24 - 5.25), mood (OR = 4.27, 95% CI: 1.96 - 9.29), order of pregnancy (OR = 3.37, 95% CI: 2.11 - 5.40), vulvitis (OR = 6.74, 95% CI: 2.66 - 17.09) and urethritis (OR = 11.02, 95% CI: 1.01 - 120.19). Vulviitis or urethritis increased 6.74 or 11.02 times the incidence of vulva dystrophy. Anger or nervous state contributed to the incidence of vulva dystrophy (OR = 4.27). Addict to hot food and order of labor also increased risk ratio for 2.55 and 3.37 times, respectively.

CONCLUSIONThe risk factors of vulvar dystrophy were: addict to hot food, often holding a angry or nervous state, increase of labors, having vulvitis and urethritis.

Diet ; Emotions ; Female ; Humans ; Multivariate Analysis ; Parity ; Risk Factors ; Vulvar Lichen Sclerosus ; etiology

OBJECTIVETo investigate the effect of Luteolin alone or combination with chemotherapentic drugs on the cytoxicity of cancer cells.

METHODSCultured A549, Hela, MCF-7, AGS, MGC-803, Caco2 and HepG2 cells were treated with Luteolin or the combination of Luteolin with other chemotherapeutic agents (Bexarotene, Cisplatin and Bleomycin). Cell viability was measured by MTS assay and IC(50) was calculated.

RESULTSThe IC(50) of Bexarotene to Hela cells was 2 micromol/L, but with the combination of 5 micromol/L of Luteolin that reduced to 0.2 micromol/L. However, the combination of Bexarotene and Luteolin did not show significant benefit in MGC-803, HepG2 cells, Caco2 and MCF-7 cells. The IC(50) of Cisplatin to Hela cells was over 30 micromol/L,but it decreased to 3 micromol/L in the presence of 5 micromol/L Luteolin; Luteolin also sensitized Cisplatin in MGC-803, HepG2 and A549 cells studied. The IC(50) of Bleomycin to Hela cells was over 100 micromol/L, but it was about 1 micromol/L in the presence of 5 micromol/L Luteolin. A549 cells were resistant to Bleomycin with an IC(50) of 100 micromol/L, 10 micromol/L Luteolin greatly enhanced the cytotoxicity of Bleomycin to the cells with the IC(50) of about 10 micromol/L. The inhibitions of MGC-803, HepG2, A549 and AGS cells didn't change by combination of Luteolin.

CONCLUSIONLow concentration of Luteolin has little toxic effect on the cancer cell lines tested in the study, but it can sensitize chemotherapeutic drugs in various cancer cell lines.

Antineoplastic Agents ; pharmacology ; Breast Neoplasms ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drug Synergism ; Humans ; Lung Neoplasms ; pathology ; Luteolin ; pharmacology ; Neoplasms ; pathology

Human placental decidua basalis-mesenchymal stem cells (PDB-MSCs) are multipotent cells from the human term placenta, which are ethically conducive, easily accessible and high-yielding source. PDB-MSCs can differentiate into adipogenic, osteogenic and neurogenic cells under appropriate conditions, which may be an attractive and alternative source of seed cells for tissue engineering. To investigate the effect of hypoxia (1% O2) on human PDB-MSCs and the expression of cytokine, PDB-MSCs were isolated from human placenta by density gradient centrifugation and cultured in the Dulbecco's modified Eagle's medium-high glucose (DMEM-HG) containing 10% fetal bovine serum (FBS), and the fifth passage of PDB-MSCs were taken. PDB-MSCs were divided into 4 groups according to the concentrations of O2 and FBS: 20% O2, 10% FBS; 20% O2, 0% FBS; 1% O2, 10% FBS; 1% O2, 0% FBS. The proliferation and apoptosis of PDB-MSCs were detected by MTT and flow cytometric analysis at the time points of 6 h, 12 h, 24 h, 48 h, 72 h, and 96 h, respectively. Vascular endothelial growth factor (VEGF) released from PDB-MSCs was detected by enzyme-linked immunosorbent assay (ELISA) at the same time points. The results showed that hypoxia enhanced the proliferation of PDB-MSCs at 12 h under the condition of 10% FBS, while at 24 h under the condition of 0% FBS (P<0.01, n=3). In normoxia, the cells cultured in 10% FBS displayed a significant proliferation compared to those cultured in 0% FBS. However, in hypoxia, the number of cells cultured in 0% FBS (serum deprivation) increased significantly compared to that cultured in 10% FBS at 24 h and 96 h respectively (P<0.05, P<0.01, n=3). With the flow cytometric analysis of cell apoptosis under the condition of hypoxia and serum deprivation, we found that hypoxia and serum deprivation did not induce PDB-MSCs apoptosis (P>0.05, n=3). This conclusion may relate to the expression of VEGF which needs further research. In conclusion, the results obtained indicate that PDB-MSCs are able to bear hypoxia and serum deprivation, suggesting that PDB-MSCs can be used as seed cells for ischemia related tissue engineering.
Apoptosis ; Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Decidua ; cytology ; Female ; Humans ; Mesenchymal Stromal Cells ; cytology ; Placenta ; cytology ; Pregnancy ; Tissue Engineering ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo rapidly and sensitively detect the four virulence-associated factors of Streptococcus suis, a multiplex PCR was developed.

METHODSIn the process of this reaction, four distinct DNA targets were amplified. One target was based on the serotype 2 (and 1/2) specific cps gene and the others were based on Streptococcus suis mrp, epf (epf*) and sly gene, encoding the MRP, EF(EF*) and Sly proteins of Streptococcus suis. 72 isolates, which including 48 strains of Streptococcus suis and 24 strains of negative control, and 49 clinical specimens were detected by the multiplex PCR assay.

RESULTSAll PCR products were detected by electrophoresis on 1.2% agarose gels. With the 48 Streptococcus suis strains, the positive detection rates of cps2+, mrp+, epf+, epf*+ and sly+ were 16/48, 14/48, 12/48, 3/48 and 26/48,respectively. The results were confirmed by bacteriological examination. There were no specific amplification products including 49 clinical specimens and 24 negative control strains.

CONCLUSIONThe results demonstrated that multiplex PCR was a highly specific and sensitive diagnostic tool for the detection of virulence-associated factors of streptococcus suis.

Bacterial Proteins ; genetics ; Polymerase Chain Reaction ; methods ; Streptococcus suis ; genetics ; pathogenicity ; Virulence Factors ; genetics

OBJECTIVETo further study the characteristics of drugs with neutral property in two-way application and conditioned dominance by observing the action characteristic of 10 traditional Chinese medicines with neutral property in hemorheological indicators of heat stagnation and blood stasis syndrome and cold stagnation and blood stasis syndrome rats.

METHODThe model of heat stagnation and blood stasis syndrome rats was established by injecting carrageenan and dry yeast, while the model of cold stagnation and blood stasis syndrome rats was established by body freezing. Subsequently, 10 traditional Chinese medicines with neutral property, 5 traditional Chinese medicines with heat property and 5 traditional Chinese medicines with cold property were selected for intervention to observe the changes in such indicators as whole blood viscosity, plasma viscosity and hematocrit and analyze the action characteristics of drugs with neutral property.

RESULTANOVA showed that among six of the 10 traditional Chinese medicines with neutral property, including Typhae Pollen, Sarcandrae Herba and Sappan lignum, could obviously increase the hemorheological indicators of both heat stagnation and blood stasis syndrome and cold stagnation and blood stasis syndrome rats; five traditional Chinese medicines with cold property, such as Salviae Miltiorrhizae Radix et Rhizoma, Leonuri Herba, Rhei Radix et Rhizoma, could significantly ameliorate the hemorheological indicators of heat stagnation and blood stasis syndrome rats (P < 0.01 or P < 0.05), and Salvia Miltiorrhiza Radix et Rhizoma alone could ameliorate the hemorheological indicators of cold stagnation and blood stasis syndrome rats (P < 0.05); all of the five traditional Chinese medicines with heat property could significantly ameliorate the hemorheological indicators of cold stagnation and blood stasis syndrome rats (P < 0.01), among which Carthami Flos and Notoginseng Radix et Rhizoma could significantly ameliorate the hemorheological indicators of cold stagnation and blood stasis syndrome rats. According to the average high-shear blood viscosity analysis, drugs with neutral property showed similar action characteristics to those with cold property in ameliorating hemorheology indicators of heat stagnation and blood stasis syndrome rat and better effect than those with heat property in reducing whole blood viscosity; and traditional Chinese medicines with neutral property have the similar action characteristics to those with heat property in improving the hemorheology indicators of cold stagnation and blood stasis syndrome rat and better effect than those with heat property in reducing whole blood viscosity.

CONCLUSIONUnder the condition of heat stagnation and blood stasis syndrome, traditional Chinese medicines with neutral property show the similar action characteristics to those with cold property; but under the condition of cold stagnation and blood stasis syndrome, traditional Chinese medicines with neutral property show the similar action characteristics to those with heat property. This indicates that traditional Chinese medicines with neutral property show both heat and cold properties under he conditions of heat stagnation and blood stasis syndrome and cold stagnation and blood stasis syndrome.

Analysis of Variance ; Animals ; Blood Circulation ; drug effects ; physiology ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Hemorheology ; drug effects ; Homeostasis ; drug effects ; Male ; Medicine, Chinese Traditional ; methods ; Rats ; Rats, Sprague-Dawley ; Thrombosis ; drug therapy ; physiopathology

OBJECTIVETo investigate the effects of a Chinese herbal extract Songyou Yin on residual hepatocellular carcinoma after chemotherapy in nude mice and the relevant mechanisms.

METHODSOrthotopic nude mouse models bearing residual hepatocellular carcinoma after chemotherapy was established using human liver carcinoma MHCC97L cells. Three different doses of Songyon Yin (2.1 g/kg, 4.2 g/kg and 8.4 g/kg) were administered to the mice in the trial groups by intragastric gavage, respectively. The mice in the control group were administered physiological saline. The tumor growth, metastasis and survival in the mice of each group were recorded. The corresponding mechanisms were studied.

RESULTSThe pulmonary metastasis rates of the control group and 2.1g/kg, 4.2g/kg, 8.4g/kg Songyou Yin treatment group were 86.7%, 73.3%, 40.0%, and 20.0%, respectively, and the survivals of these groups were 53.83 ± 4.71, 56.50 ± 6.09, 66.67 ± 5.61, 81.17 ± 7.36 days, respectively. Compared with the mice in the control group, mice in the 4.2 g/kg, 8.4 g/kg Songyou Yin treatment groups had a lower pulmonary metastasis rate (P = 0.021 and P = 0.001, respectively) and longer survival (P = 0.002 and P = 0.001, respectively). A restoration of E-cadherin expression and a concomitant reduction of N-cadherin expression were detected in the tumors of the 4.2 g/kg and 8.4 g/kg Songyou Yin treatment groups.

CONCLUSIONSSongyou Yin effectively inhibits the invasion and metastasis of the residual hepatocellular carcinoma after chemotherapy in nude mice through attenuating the epithelia-mesenchymal transition and prolongs the survival. Songyon Yin may have potential to promote the efficacy of chemotherapy in hepatocellular carcinoma.

Animals ; Antineoplastic Agents ; therapeutic use ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Cadherins ; metabolism ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; pathology ; Cell Line, Tumor ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Epithelial-Mesenchymal Transition ; drug effects ; Humans ; Liver Neoplasms ; drug therapy ; metabolism ; pathology ; Lung Neoplasms ; secondary ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neoplasm, Residual ; metabolism ; pathology ; Organoplatinum Compounds ; therapeutic use ; Plants, Medicinal ; chemistry ; Random Allocation ; Survival Rate ; Tumor Burden ; drug effects ; Xenograft Model Antitumor Assays