Objective To investigate the effects of electroacupuncture (EA) on affected and healthy limbs and on the regulation of insulin-like growth factor-1 (IGF-1) mRNA and proteins in the cerebral cortex early after focal ischemic.Methods A model of focal cerebral ischemia was established in 54 SD rats using the suture occlusion method.They were then randomly divided into an affected limb therapy group (ALTG,n =18),an unaffected limb therapy group (UALTG,n =18),and a control group (CG,n =18).Each group had a 7-day subgroup,a 14-day subgroup and a 21-day subgroup with 6 rats in each.Rats in the experimental groups received EA beginning 24h after the occlusion.Rats in each subgroup were sacrificed in a random order on the 7th,14th and 21st days and the ischemic cerebral cortexes were quickly dissected.The specimens were frozen in liquid nitrogen before being analysed for IGF-1 mRNA expression by RT-PCR and for IGF-1 protein by Western blotting.Results ①After occlusion,IGF-1 protein levels in the ischemic cortexes of the CG declined from the 7th through the 21st day.Rats in the ALTG had significantly higher levels compared with the CG at all time points.The UALTG had the highest values on the 14th day,but was lower than the ALTG and higher than the CG at the 21st day.②IGF-1 mRNA levels in the ischemic cortexes of the UALTG declined from the 7th through the 21st day.At day 7 the results of the UALTG were 6.8 times higher than the CG,and the ALTG was 3.0 times higher.At day 14 levels in the UALTG were significantly lower than those in the ALTG.At that point the results of the UALTG rats were 3.3 times higher than those of the CG and the ALTG was 5.7 times higher.On day 21 levels in both the UALTG and ALTG were significantly lower than in the CG.Conclusions EA intervention at an early stage of focal cerebral ischemia can improve the expression of IGF-1 mRNA and protein levels in the ischemic cortex.Treating the unaffected limb can evoke more IGF-1 mRNA expression earlier and with relatively longer duration,and generate relatively longer protein increases.EA administered to the unaffected limb was more effective in the early stage of stroke.

An experiment was conducted using cultivated Glycyrrhiza uralensis in age of one year to study the effects of abscisic acid (ABA) on chemical components content and color of G. uralensis. By using different concentrations of ABA spraying on leaves, the change of the chemical component content was analyzed within 45 d after ABA stimulation, and the effects on quality were studied combined with colorimetric analysis data. It turned out that in some sense the content of glycyrrhizic acid and liquiritin had increased within 45 d, especially for liquiritin. After high concentrations of ABA (3.96 mg · L(-1)) stimulating, the content of glycyrrhizic acid rose 52% while liquiritin up 392% within 30 d. Then they both showed a decline in the content of glycyrrhizic acid and liquiritin on 45 d. Color index values of a* and b* were all significantly higher than that of the control group within 45 d, which meant the color of powders turned toward red and yellow. The conclusion was that ABA (3.96 mg · L(-1)) stimulating could not only improve the quality in the traditional sense through the color of G. uralensis, but also in the modern sense by improving the content of glycyrrhizic acid and liquiritin.
Abscisic Acid ; pharmacology ; Color ; Drugs, Chinese Herbal ; chemistry ; Flavanones ; analysis ; Glucosides ; analysis ; Glycyrrhiza uralensis ; chemistry ; drug effects ; growth & development ; Glycyrrhizic Acid ; analysis ; Plant Growth Regulators ; pharmacology

OBJECTIVETo investigate the effect of compound Coptidis Rhizoma capsule (CCRC) on unbalanced expression of renal tissue TGF-β1/BMP-7 and Smad signaling pathway in rats with early diabetic nephropathy (DN), and discuss CCRC's effect on DN rats with early diabetic nephropathy and its possible mechanism.

METHODDN model rats were established by injecting streptozotocin (STZ). The rats were randomly divided into seven groups: the normal group, the model group, the enalapril treatment group, the xiaoke pill treatment group and three CRCC treatment groups. They were orally administered once a day for five weeks. The fasting blood glucose (FBG), blood urea nitrogen (BUN), serum creatinine (Scr), insulin (Ins), 24 h urinary protein (24 h Upro) and 24 h urinary microalbumin (24 h UmAlb) were tested. The pathological changes in renal tissues were examined by optical microscopy. Immuno- histochemical measures were used to detect the expressions of TGF-β1, BMP-7, Smad2/3, Smad1/5, and Smad7 protein, and RT-PCR was used to detect TGF-β1 mRNA and BMP-7 mRNA in renal tissues.

RESULTCompared with model group, BUN, Scr, Ins, 24 h Upro and 24 h UmAlb levels decreased at different degrees in CCRC treatment groups; the abnormal pathomorphology in renal tissue was improved; immunohistochemistry results showed that the expression of TGF-β1 and Smad2/3 were reduced, while the expression of BMP-7, Smad1/5 and Smad7 increased in CRCC treatment groups; the expression of TGF-β1 mRNA were reduced, but the expression of BMP-7 mRNA had no obvious change in CRCC treatment groups.

CONCLUSIONCRCC can improve the early renal function, delay the progression of chronic renal pathology and maintain the dynamic balance of TGF-β1/BMP-7 expression in renal tissues of DN rats. The mechanism may be related to down-regulation of renal TGF-β1 and up-regulation of BMP-7 through Smad signaling pathway.

Animals ; Bone Morphogenetic Protein 7 ; genetics ; metabolism ; Coptis ; chemistry ; Diabetic Nephropathies ; drug therapy ; genetics ; metabolism ; Gene Expression Regulation ; drug effects ; Humans ; Kidney ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Rhizome ; chemistry ; Signal Transduction ; drug effects ; Smad Proteins ; genetics ; metabolism ; Transforming Growth Factor beta1 ; metabolism

Aim To observe effect of DADS on the differentiation of gastric adenocarcinoma cells and acetylation of human gastric cancer tumor cell transplantable histone.Methods Human gastric adenocarcinoma heterotransplantable tumor model was constructed through subcutaneously injecting MGC803 cells to nude mice.Morphologic changes of xenograft tumor cells were observed with optical microscope,the influence of DADS on xenograft tumor cells generationcycle distribution,the expression of p21~(WAF1) protein,histone H3 and and H4 acetylion were analyzed with flow cytometry and Western blot.Results There was obvious growth inhibitory effect of xenograft tumor while abdominal injection dose were 100 and 200 mg?kg~(-1)DADS;cells density and heteromorphism decreased after treated with DADS.Flow cytometry analysis revealed that treating xenograft tumor cells with increasing quantities of DADS increased the percentage of cells in the G_2/M phase.The proportion of xenograft tumor cells in the G_2/M phase after treatment with 100 mg?kg~(-1) and 200 mg?kg~(-1) DADS was 2.22 and 3.37 times of that in NS group.Western blot analysis showed H3 acetylion increase along with G_2/M arrest of xenograft tumor cells by DADS.DADS didn′t influence the expression level of H4 acetylion;the expression of p21~(WAF1) protein in xenograft tumor increased along with the increases in the concentration of DADS.Conclusion DADS cansignificantly inhibit the growth of human gastric carcinoma xenograft in BALB/C nucle mice and induce cell differentiation,which might be related with up-regulation of histone a cetylization and p21~(WAF1) protein level.

Objective To investigate the functional and histological changes in aging gerbil bladders.MethodsTwelve male gerbils were randomly divided into control group and aging bladder group,with 6 in each group.Experiment was conducted in gerbils of control group and aging bladder group 6 months and 24 months after normal feeding,respectively.Urodynamic examinations including irrigation volume,compliance and filling pressure of bladder were performed,bladder weight was obtained,bladder weight index was calculated,morphological observation was conducted with HE staining,ratio of amount of smooth muscle to collagen was obtained with double immunohistochemical staining,and electron microscope was used to evaluate the ultrastructure of bladder.ResultsCompared with those in control group,the irrigation volume and compliance of bladder significantly decreased in aging bladder group(P0.05).However,the bladder weight index was significantly lower in aging bladder group(P0.05).There existed degeneration in smooth muscle cells and organelles in aging bladder group.ConclusionThe function of aging bladder in gerbils is impaired,which may be related to the degeneration of bladder tissues.

OBJECTIVETo compare the intervention effects of four traditional Chinese medicines (TCMs) with typical cold or hot property on body temperature and temperature-sensitive transient receptor potential ion channel proteins (TRPs) of rats with yeast-induced fever.

METHODThe pyrexia model was induced by injecting yeast suspension subcutaneously. Totally 108 male SD rats were randomly divided into the normal group, the model group, the Rhei Radix et Rhizoma treated group, the Coptidis Rhizoma treated group, the Euodiae Fructus treated group, and the Alpiniae Officinarum Rhizoma treated group, with 18 rats in each group. At the 4 h, 8 h and 12 h after injection of yeast, the rats were sacrificed to collect their hypothalamus and dorsal root ganglion. The expressions of TRPV1 and TRPM8 were detected by immunohistochemistry and Western blot method.

RESULTCompared with the normal group, after injection of yeast, the temperature of rats in the model group notably increased, and reached the peak at 8 h (P < 0.01). The TRPV1 level in hypothalamus and dorsal root ganglia (DRG) of the model group significantly increased, whereas the TRPM8 level significantly reduced. Compared with the model group, the Rhei Radix et Rhizoma group and the Coptidis Rhizoma group showed significant decrease in the high body temperature of rats caused by yeast, down-regulation in the expression of TRPV1, and up-regulation in the expression of TRPM8 (P < 0.05 or P < 0.01). Euodiae Fructus and Alpiniae Officinarum Rhizoma had no significant effect on either temperature or TRPs of fever rats.

CONCLUSIONRhei Radix et Rhizoma and Coptidis Rhizoma, both are TCMs with cold property, can reduce the temperature of fever rats induced by yeast, which may be related to their effective regulation of TRPV1 and TRPM8 in hypothalamus and DRG, while Euodiae Fructus and Alpiniae Officinarum Rhizoma had no relevant effect.

Animals ; Antipyretics ; administration & dosage ; chemistry ; Body Temperature Regulation ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Fever ; drug therapy ; immunology ; microbiology ; physiopathology ; Gene Expression Regulation ; drug effects ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Saccharomyces cerevisiae ; immunology ; TRPM Cation Channels ; genetics ; immunology ; TRPV Cation Channels ; genetics ; immunology

The mechanism of biological actions of quercetin was studied by using metabolomic method and biomolecular network. HPLC-MS was used to analyze the serum metabolome in rats of blank group and quercetin administration group rats, and MS data were processed by MATLAB software. With multivariate statistical analysis of serum metabolite profiles, a clear separation among blank group and quercetin administration group was achieved, potential biomarkers were selected according to the parameters of variable importance in the projection (VIP) and identified according to MS information and database retrieval. Four compounds, related enzymes, action targets and metabolic pathways had been confirmed, namely retinoic acid and RARbeta, arachidonate and COX-2, 3, 5-diodotyrosine and TPO, uridine diphosphate glucose and PDEs. The mechanism of quercetin enhancing ability of retinoic acid on the induction of RARbeta, activating TPO, using as COX-2 and PDEs inhibitor was approved by biomolecular network and related literatures. In this study, a mechanism of multiple biological actions of quercetin was evaluated at the level of the biomolecular network, metabolomics and biomolecular network can be used to investigate the biological effects mechanism of quercetin, which provided a new method to further revealing mechanism of drug action.
Animals ; Antioxidants ; pharmacology ; Biomarkers ; blood ; Chromatography, Liquid ; Mass Spectrometry ; Metabolic Networks and Pathways ; Metabolome ; Metabolomics ; Multivariate Analysis ; Quercetin ; pharmacology ; Rats

The aim of the manuscript was to optimize formulations and preparation technologies of cataplasm of white mustard seed varnish, and to evaluate its anti-asthma effect on rats. The single factor experiments included spreading thickness, types of crosslinking agents, dihydroxyaluminum aminoacetate amount, sodium polyacrylate amount, types of adhesive agents with human sense as the evaluation index. Blank cataplasm matrix was optimized by the orthogonal experiment with the amount of glycerine, citric acid, and sodium carboxymethylcellulose as the major influential factors. Initial adhesive force, peeling strength and human sense were as the evaluation index. The optimized formulation of blank cataplasm were as followings: glycerine-water-ethanol-PEG400-dihydroxyaluminum aminoacetate-citric acid-sodium carboxymethylcellulose-sodium carboxymethylcellulose 2 : 8 : 0.8 : 0.4 : 0.07: 0.15 : 0.1 : 0.5. The active ingredients of white mustard seed, corydalis, and gansui root were extracted by alcohol extraction method. Asiasarum volatile oil was extracted by oil extractor. The optimized drug loading amount was 11% with initial adhesive force, peeling strength and human sense as the evaluation index. Asthma rats model were established by sensitized with ovalbumin and nose-scratching time as the evaluation index. High dose (17%) group of drug-loaded cataplasm had the obvious inhibition effect on nose-scratching time of rats (P = 0.037 < 0.05). In comparison, middle dose (11%), low dose (4%) and positive-control groups had no obvious inhibitive effect on rats. White mustard seed cataplasm supplied a novel choice for anti-asthma therapy. And the overall pharmacodynamics assessment will be carried out on molecular level in near future.
Animals ; Anti-Asthmatic Agents ; administration & dosage ; chemistry ; Asthma ; drug therapy ; Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Female ; Humans ; Male ; Mustard Plant ; chemistry ; Rats ; Rats, Sprague-Dawley ; Seeds ; chemistry

The fermentative H2-producing strain Clostridium sp. H-61 was isolated from anaerobic sludge,was used as an original strain which was induced by NTG and UV for increasing and the hydrogen production ability. One of the highest efficient H2-producing mutants was named as HCM-23 with its stable hydrogen production ability. which was measured in the batch culture experiments. With the condition of 10 g/L glucose,its cumulative hydrogen yield and hydrogen production rate was 3024 mL/L and 33.19 mmol H2/g DW?h,69.89% and 68.14% higher than that of the original strain,respectively. The terminal liquid product compositions showed that the mutant HCM-23 fermentation was ethanol type,while the original strain H-61 fermentation was butyric acid type. Varieties of parameters of hydrogen production fermentation studied,including time,carbon source,nitrogen source,glucose concentration,glucose utilization,initial pH and incubation temperature had been studied,indicated the optimum condition of hydrogen production for the mutantHCM-23 as initial pH 6.5,temperature 36 ℃,and the favorite substrate was sucrose. The hydrogen production characters of the mutant and the original strain were different,such as,the growth lag phase and the utilization of inorganic nitrogen source,etc. This work shows a good application potential of NTG-UV combined mutation in the biohydrogen production. And the hydrogen production mechanism and metabolic pathway should be explored furthermore.

AIM: To explore the potential of human bone marrow mesenchymal stem cells (hMSCs) differentiating into hematopoietic cells with murine fetal liver stromal cell-conditioned medium (FLSC-CM) in vitro. METHODS: 12.5-14.5 days post coitus (dpc ) of KM mice were used for the preparation of fetal liver stromal cell-conditioned medium (FLSC-CM) and embryonic fibroblast feeder layer (FD). Culture-expanded hMSCs were directly contacted with FLSC-CM, FD, and the combination of human interleukin-6 (IL-6) or stem cell factor (SCF), respectively. Seven days later, the non-adherent cells were collected and characterized by morphology, immunophenotypes, and colony forming unit-granulocyte/macrophage culture assay. RESULTS: The number of nonadherent cells derived from hMSCs cultured with FLSC-CM was increased remarkably than those with either FD or cytokines. The non-adhered cells with the morphology of monocyte-or small lymphocyte-like cells were positive for human CD34, CD45 and had the capacity to form the hematopoietic progenitor colonies in methylcellulose cultures containing recombined human granulocyte/macrophage-colony stimulating factor (rhGM-CSF). CONCLUSION: hMSCs were successfully induced toward their differentiation into CD34+CD45+ hematopoietic progenitors after being cultivated with FLSC-CM. This study suggests that hMSCs have the hematopoietic differentiation potential in vitro. [