High performance liquid chromatography-time-off-flight mass spectrometer (HPLC-TOFMS) technology coupled with partial least squares discriminant analysis (PLS-DA) processed by SIMCA-P software was applied to investigate serum endogenous metabolites alternations of valsartan in spontaneous hypertension rats (SHR). And MetPA platform was used to connect identified potential biomarkers in corresponding metabolic pathways to find possible therapeutic mechanism of valsartan. Valsartan significantly declined the blood pressure of SHRs (P < 0.05) at fourth week. The metabolic profiling significantly changed and four metabolites involved in G protein-coupled pathway were identified. Metabolomics is able to detect holistic and microcosmic alternations in organism, so as to elucidate therapeutic mechanism of drugs.
Animals ; Biomarkers ; blood ; Blood Pressure ; Chromatography, High Pressure Liquid ; Discriminant Analysis ; Least-Squares Analysis ; Mass Spectrometry ; Metabolome ; Metabolomics ; Rats ; Rats, Inbred SHR ; Valsartan ; pharmacology

flow cytometry.A strong linear relationship was observed in the standard curve of real-time PCR of each bacteria. Conclusion These three non-culture methods can be used in the quantitative analysis of oral microorganisms.Real-time PCR and laser scanning confocal microscopy are better than the traditional culture-based CFU count,and real-time PCR is the most sensitive method.

Objective To detect the differential gene expression between Streptococcus sobrinus(S.sobrinus) 6715 and its fluoride-resistant strains. Methods The fluoride-resistant strains of S.sobrinus 6715 was induced by increasing the concentration of fluoride step by step.Total RNA of both S.sobrinus 6715 and its fluoride-resistant strains was extracted,mRNA was separated and purificated,and then cDNA was obtained by reversed transcription.Suppression subtractive hybridization(SSH) technology was used to detect the differential gene expression between them.The differential gene expression fragments were cloned and compared with the GenBank by BLAST.Results After comparing with the GenBank by BLAST,it was identified that there were two differential gene expression fragments,fruA and SMU.438c. Conclusion The cDNA subtractive lib of differential gene expression between S.sobrinus 6715 and its fluoride-resistant strains was successfully constructed through SSH,which paves a way for the further study of fluoride-resistant mechanism.

8).The amount of the targeted microorganisms(Streptococcus mutans,Streptococcus sobrinus,Actinomyces viscosus and Actinomyces naeslundii) and the total number of bacterial cells were determined by real-time PCR based on SYBR-Green I fluorescence.Results The percentages of the four targeted bacteria in high-caries group were significantly higher than those in caries-free group(P

Thirty SHRs were obtained randomly to hypertension, model group, captopril group and Qingre jiangya capsule group. Ten Wistar rats were used as control group. The hippocampus tissue was removed to explore the damage of spontaneously hypertensive rats (SHR) and the protective effect of Qingre jiangya capsule after continuously administered for 14 days. And then the data were processed by principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA). The research results revealed captopril group was significantly different from the other three groups. The classification of other three groups is also very clear after captopril group removed. This suggested that Qingre jiangya capsule could improve the overall metabolism compared with captopril. Four metabolites were identified: dimethylglycine, glycerophosphocholine, aldosterone and noradrenaline. Hypertension hippocampus damage may mainly be expressed in tyrosine metabolism, aldosterone-regulated sodium, vascular smooth muscle contraction reabsorption, and Qingre jiangya capsule could reverse the hippocampus tissue damage of SHR.
Animals ; Capsules ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Hippocampus ; drug effects ; Hypertension ; drug therapy ; Male ; Rats ; Rats, Inbred SHR ; Rats, Wistar

OBJECTIVETo study the changes of urine metabolites in hypertension patients of ascendant hyperactivity of Gan yang syndrome (AHGYS), and to explore its essence in hypertension patients.

METHODSTen typical hypertension patients of AHGYS were recruited as the patient group, and the other twelve healthy volunteers were recruited as the normal group. The metabolite profiling in the urine were collected using by high performance liquid chromatography coupled with time of flight mass spectrometry (HPLC-TOFMS). The principal component analysis (PCA) and partial least-square discriminant analysis (PLS-DA) were analyzed using SIMCA-P Software. The differential metabolites in the urine were found out and identified. The possible relevant metabolic pathways were explained.

RESULTSThe data from the analysis by PCA in the urine samples of the patient group and the normal group showed, two sets of data could be obviously classified in the score plot. Compared with the normal group, significant changes happened to the body metabolism in the patient group. The metabolites relevant to hypertension patients of AHGYS were determined using the PLS-DA. Fifteen compounds of the structure and metabolic pathways had been confirmed through inquiring KEGG Database, mainly including amino acids, free fatty acids, sphingosine, and so on.

CONCLUSIONSThe hypertension patients of AHGYS were studied using HPLC-TOFMS combined with pattern recognition, thus finding out small molecular metabolic markers from the microscopic field, which was advantageous in probing the biological nature of Chinese medicine syndromes.

Adult ; Aged ; Case-Control Studies ; Chromatography, High Pressure Liquid ; methods ; Discriminant Analysis ; Female ; Humans ; Hypertension ; diagnosis ; urine ; Least-Squares Analysis ; Male ; Mass Spectrometry ; methods ; Medicine, Chinese Traditional ; methods ; Metabolome ; Middle Aged ; Principal Component Analysis

OBJECTIVETo realize quadratic formula optimization of Renshen Jianxin Capsule (RJC) by screening Chinese herbs with major anti-myocardial ischemia effect in RJC and optimize their optimal dosages.

METHODSBy following "uniform design-pharmacodynamic experiment-mathematical modeling-formula optimization", authors employed U10(10(8)) uniform design in the experiment. Eight Chinese herbs contained in RJC were taken as observatory factors. Electrocardiograph (ECG) changes of myocardial ischemia induced by isoproterenol were taken as pharmacodynamic indices. The mathematical model between herbal factors and pharmacodynamic indices was established using stepwise regression analysis to screen Chinese herbs with major anti-myocardial ischemia effect. Their optimal dosages were optimized using the grid algorithm.

RESULTSThe regression equation was y =1. 7889 -0. 3247 Ginseng xSalvia Miltiorrhiza -0. 0663 Astragalus membranaceus xOriental Waterplantain tuber. Forecasting factors included were Ginseng, Salvia Miltiorrhiza, Astragalus membranaceus, and Oriental Waterplantain tuber. The optimal formula dosage calculated by the grid algorithm was Ginseng 1. 62 g, Astragalus membranaceus 4. 62 g, Salvia Miltiorrhiza 2. 43 g, and Oriental Waterplantain tuber 1. 66 g.

CONCLUSIONUniform design combined with stepwise regression analysis and grid algorithm were able to realize quadratic formula optimization of RJC.

Astragalus membranaceus ; Chemistry, Pharmaceutical ; standards ; Coronary Artery Disease ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Electrocardiography ; Humans ; Isoproterenol ; Myocardial Ischemia ; drug therapy ; Panax ; Salvia miltiorrhiza

OBJECTIVETo test the effects of autologous skeletal myoblast transplantation on the hemodynamics in dogs with coronary microembolization-induced chronic heart failure (CHF).

METHODSCHF models were successfully induced in 19 dogs and divided into ASMT group (n=9) and control group (n=10). The myoblasts were injected into the embolized region in the 9 dogs of the ASMT group, and saline was injected in the control dogs, and the hemodynamics of the dogs were evaluated 5 weeks after the injections.

RESULTCompared with saline injection, ASMT significantly increased dP/dtmax, MAP and LVSP (P<0.05) and decreased LVEDP (P<0.05) 5 weeks after myoblast transplantation. Desmin and Brd-U immunofluorescent staining showed myoblast survival at the injected sites in the dogs.

CONCLUSIONASMT provides mild improvements in the hemodynamics of dogs with CHF.

Animals ; Chronic Disease ; Dogs ; Female ; Heart Failure ; physiopathology ; therapy ; Hemodynamics ; Male ; Myoblasts, Skeletal ; transplantation ; Transplantation, Autologous

BACKGROUNDThe efficiency of traditional cryopreservation of human embryonic stem (ES) cells is low, and there have been few attempts to prove new cryopreservation methods effective. This study was designed to evaluate the efficiency of cryopreservation of human ES cells using vitrification method.

METHODSHuman ES cells clumped from an identical cell line were randomly allocated to be cryopreserved by vitrification or by slow freezing. The recovery rates, the growth and differentiation potential of thawed human ES cells were compared between these two groups. The pluripotency of human ES cells after thawing was identified.

RESULTSEighty-one point nine percent (59/72) of human ES cell clumps were recovered after vitrification, while only 22.8% (16/70) were recovered after slow freezing (P < 0.01). The colonies after vitrification manifested have not only faster growth but also a lower level of differentiation when compared to colonies subjected to the slow freezing protocol. However, the rates of growth and differentiation in undifferentiated colonies from both groups were identical to the rates in those of non-cryopreserved stem cells after a prolonged culture period. Passage 6 of vitrified human ES cells retained the properties of pluripotent cells, a normal karyotype and expressed the transcription factor OCT-4, stage specific expressed antigen-4 (SSEA-4) and SSEA-3. Teratoma growth of these cells demonstrated the ability to develop into all three germ layers.

CONCLUSIONSVitrification is effective in cryopreserving human ES cells. During a prolonged culture, human ES cells retain their pluripotency after cryopreservation.

Cell Differentiation ; Cell Survival ; Cryopreservation ; methods ; Embryo, Mammalian ; cytology ; Humans ; Osmotic Pressure ; Stem Cells ; cytology

BACKGROUNDHuman embryonic stem (HES) cell derived from human blastocyst can be propagated indefinitely in the primitive undifferentiated state while remaining pluripotent. It has exciting potential in human developmental biology, drug discovery, and transplantation medicine. But there are insufficient HES cell lines for further study.

METHODSThree oocyte donors were studied, and 3 in vitro fertilization (IVF) cycles were carried out to get blastocysts for the establishment of HES cell line. Isolated from blastocysts immunosurgically, inner cell mass (ICM) was cultured and propagated on mouse embryonic fibroblasts (MEFs). Once established, morphology, cell surface markers, karyotype and differentiating ability of the cell line were thoroughly analyzed.

RESULTSFour ICMs from 7 blastocysts were cultured on MEFs. After culture, one cell line (cHES-1) was established and met the criteria for defining human pluripotent stem cells including a series of markers used to identify pluripotent stem cells, morphological similarity to primate embryonic stem cells and HES reported else where. Normal and stable karyotype maintained over 60 passages, and demonstrated ability to differentiate into a wide variety of cell types.

CONCLUSIONSHES cell lines can be established from gamete donors at a relatively highly efficient rate. The establishment will exert a widespread impact on biomedical research.

Blastocyst ; cytology ; Cell Differentiation ; Cell Line ; DNA-Binding Proteins ; analysis ; Female ; Fertilization in Vitro ; Humans ; Karyotyping ; Male ; Octamer Transcription Factor-3 ; Stem Cells ; cytology ; Tissue Donors ; Transcription Factors ; analysis