Objective To study the characteristics of pharmacokinetics and pharmacodynamics of insulin dry powder inhalation and its relative bioavailability as compared with subcutaneous injection of regular insulin. Methods In this open,single-center,randomized,two-period,cross-over,euglycemic glucose clamp study,18 healthy volunteers(14 men and 4 women),aged(24.9?1.7)years,with body mass index(20.6?1.2)kg/m~2, received the insulin dry powder inhalatin(80 U)or regular insulin(15 U)subcutaneous administration.The blood samples of this study at 0,20,30,40,50,60,70,80,90,100,110,120,135,150,165,180,195, 210,225,240,270,300,330,360,390,420,450 and 480 rain were taken for serum insulin measurement, meanwhile,glucose infusion rates(GIR)were determined per 5 minutes over a period of 8 hours.Results The C_(max)were(57.9?17.8 vs 114.5?29.7)mU/L(tested vs reference preparation),T_(max)were(46.7?45.6 vs 107.8?33.7)min,GIR_(max)were(3.35?0.98 vs 5.17?1.75)mg?kg~(-1)?min~(-1)and T_(GIRmax)were(88.3?17.0 vs 151.9?34.6)min.The relative bioavailability was(10.26?2.25)%,and the relative bioefficacy was(14.33?7.26)%.Conclusion The study shows that insulin dry powder inhalation is absorbed via lungs and its action sets in earlier than that of the regular insulin injected subcutaneously.These pharmacokinetie and pharmacodynamic data may provide a reliabe guide for further clinical trial.

Objective To investigate whether catalpol is the active component responsible for the Yin tonic effect of Radix Rehmanniae.Methods Young NH mice were injected with triiodothyronine to produce the hyperthyroidism model,while old mice were used as the model of natural aging.The single point radioligand binding assay was carried out to determine the ?-adrenergic receptor density and M-cholinergic receptor density.The learning ability(short term memory) was determined by the Y-maze avoidance test. Results In the ?-adrenergic receptor experiment,the densities were(15.7?5.2) and(20.9?7.2) fmol/mg protein in normal control group and in T3 control group(P

Objective To verify the association between common breast cancer susceptibility loci which have been confirmed in European and Asian populations and breast cancer susceptibility in sporadic breast cancer among the Han nationality in Henan province , and to analyze their genotypes in the internal type of breast cancer . Methods In 253 breast cancer patients ( the case group ) and 343 patients who had benign breast lesions ( the control group), rs2046210(6q25.1), rs2981582(EGFR2), rs889312(MAP3K1), and rs3803662(TOX3/TNRC9)were genotyped by SNP im-LDR technique.According to estrogen receptor(ER), progesterone receptor (PR), human epidermal growth factor receptor 2(HER2)and Ki67, breast cancer are divided into 5 types:Lu-minal A, Luminal B, HER2-enrich, Luminal HER2, and triple negative breast cancer ( TNBC).Results rs2046210(6q25.1), rs2981582(EGFR2), rs889312(MAP3K1)had no statistical differences between the case group and the control group(P=0.421, 0.459, and 0.468), but the genotype of rs3803662(TOX3/TNRC9)in the case group and the control group had statistical difference (P=0.037).The allelic frequencies of rs3803662 between the case group and control group were different in codominant inheritance ( OR=2.19, 95%CI:1.19-4.02)and recessive genetic models (OR =2.06,95% CI:1.15 -3.70).Compared with AA and GA, GG in-creased the risk of breast cancer ( P =0.012, 0.015 ).The genotypes of rs2046210 ( 6q25.1 ), rs2981582 (EGFR2), rs889312(MAP3K1), and rs3803662(TOX3/TNRC9)had no difference in different types of breast cancer.Conclusions Four common breast cancer susceptibility loci from GWAS are not entirely associated with breast cancer risk among the Han nationality in Henan province .Only rs3803662(TOX3/TNRC9)is confirmed to increase the risk of breast cancer .Different genotypes of 4 loci distribute equally in different types of breast cancer .

The cDNA encoding human Vascular Endothelial Growth Factor 165 (VEGF165) was amplified using RT-PCR from human tonsil tissue and cloned into eukaryotic expression vector pcDNA3.1 (+). The recombinant plasmid pcDNA/V was transferred into 293 cells mediated by liposome and the cells stably expressing VEGF were selected under the pressure of G418. ELISA and Western blotting demonstrated that the eukaryotic expression vector pcDNA/V was successfully constructed and its corresponding protein could be expressed efficiently in vitro. Chick Charioallantoic Membrane (CAM) bioassay showed that recombinant protein has biological activity of hVEGF. Model rats with acute myocardial ischemia were used to further study the expression of VEGFin vivo. The model rats were divided randomly into three groups: control group, pcDNA3.1 (+) group and pcDNA/V group. 50microL naked plasmid DNA or saline was intramyocardially injected at three sites into the border zone of infarction. The hearts of rats were excised and fixed histologically, then the infarction sizes were studied by immunohistochemical staining and electron microscope after four weeks. Immunohistochemical staining for VEGF appeared to be negative in control and pcDNA3.1 (+) groups. In pcDNA/V group, myocardial cells in infarction border zone showed positive staining for VEGF in cytoplasm. Ultrastructural anaylsis showed that there were visible hyperplasia of vascular endothilium in pcDNA/V group. The control and pcDNA3.1 (+) groups showed less capillary hyperplasia. In this study, VEGF165 gene was successfully cloned and its protein expressed in vitro and in vivo was of bioactivity, which provides a basis for the further study of biological functions of human VEGF.
Animals ; Cell Line ; Chickens ; Chorioallantoic Membrane ; blood supply ; Disease Models, Animal ; Genetic Therapy ; Humans ; Male ; Myocardial Infarction ; metabolism ; pathology ; therapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; biosynthesis ; genetics ; therapeutic use ; Transfection ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

Objective To study the relationship between facial skin conditions and TCM constitutional types; To provide new means and methods for guidance of skin conservation and skin health improvement from the aspect of constitutional types. Methods Totally 2241 female volunteers in nine provincial capital cities and municipalities participated in this research, and were investigated by web questionnaire. Constitutional types were determined according to the“Classification and Judgment of TCM Constitutional Types”standard issued by China Association of Chinese Medicine. Skin color, glossiness, delicate degree, moistness, firmness, resilience, uniformity and other indicators were scored. Skin conditions were classified into three groups:1-3 score group, 4-6 score group, and 7-9 score group, and the constitutional differences among each group were compared. Results Yang-deficiency and mild constitutional types accounted for the largest proportion. Yellowish skin color accounted for the largest proportion, and the next were white, dark and reddish. Yang-deficiency constitutional type accounted for the largest proportion in 1-3 score group, and the second was Qi-stagnation; Yang-deficiency and mild constitutional types accounted for the largest proportion in 4-6 score group; mild constitutional type accounted for the largest proportion in 7-9 score group. Conclusion People with mild constitutional type got highest score in every skin index, while biased constitutional types influenced the score in varied degrees. The constitutional types which influenced Chinese urban females’ facial skin index more were Yang-deficiency and Qi-stagnation constitutional types.

OBJECTIVETo study the rehabilitation effects ergometry on COPD patients.

METHODSThirty COPD out-patients in our Hospital were randomly divided into 2 groups. Rehabilitation group, 15 patients, performed leg ergometry exercise of 80% peak Watt x 30min/d x 3d/w x 12w. Another 15 patients were control group without exercise. All patients received conventional therapy. Pulmonary function testing (PFT), cardiopulmonary exercise testing (CPET), arterial blood gas analysis (ABG), Borg and CAT sores were done at both baseline and 12 w.

RESULTSThere was no statistically difference in lung function testing, blood gas analysis and cardiopulmonary exercise test when pre- exercises between 2 sub-groups. The IC, peak VO2 and peak, W of rehabilitation group significantly increased (P < 0.05); and Borg and CAT.scores significantly decreased (P < 0.05) from baseline; and other PFT and ABG did not change (P > 0.05). While there was no difference in control group (P > 0.05).

CONCLUSIONLeg submaximal ergometry rehabilitation improves health condition and ameliorate dyspnea symptoms in COPD patients.

Blood Gas Analysis ; Dyspnea ; therapy ; Exercise Test ; Exercise Therapy ; Humans ; Pulmonary Disease, Chronic Obstructive ; therapy ; Respiratory Function Tests

OBJECTIVETo observe the change of PTEN/PI3K/AKT pathway hypoxia-inducible factor (HIF-1α), vascular endothelial growth factor (VEGF) in rats with adjuvant arthritis and to explore the mechanism of neovasculization in rheumatoid arthritis.

METHODSThirty rats were randomly divided into normal control group and model control group. The model control group were established the model of adjuvant arthritis using Freund's complete adjuvant. At 19 days after modeling, the expression of microvascular density (MVD), HIF-1α, VEGF were detected by ELISA assay and PTEN, PI3K, AKT were detected by Werstern Blotting.

RESULTSCompared with the normal control group, paw swelling, arthritic index were increased, and the expression of MVD, VEGF, HIF-1α of serum, PI3K, AKT of synovial tissue were significantly increased, PTEN was significantly decreased in model control group. PI3K, HIF-1α were positively correlated with MVD; VEGF, AKT were positively correlated with paw swelling; PTEN was negatively correlated with the arthritis index; HIF-1α was positively correlated with VEGF; PI3K was positively correlated with AKT, PTEN was negatively correlated with PI3K, AKT, VEGF.

CONCLUSIONImbalance of PTEN/PI3K/AKT pathway in rats with adjuvant arthritis is one of the mechanisms of synovial neovasculization.

Animals ; Arthritis, Experimental ; physiopathology ; Hypoxia-Inducible Factor 1, alpha Subunit ; physiology ; Neovascularization, Pathologic ; etiology ; PTEN Phosphohydrolase ; physiology ; Phosphatidylinositol 3-Kinases ; physiology ; Proto-Oncogene Proteins c-akt ; physiology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; physiology

OBJECTIVETo investigate the effect of 6-gingerol, the main active component of ginger, on hair shaft elongation in vitro and hair growth in vivo.

METHODSFirstly, Hair follicles were co-cultured with 3 different concentration of 6-gingerol for 5 days and hair elongation in three groups was measured. Secondly, The proliferative effect of 6-gingerol on DPCs was measured using MTT assay. Thirdly, the expression of Bcl-2 and Bax in DPCs were measured using Western blotting. In vivo study, the influence of 6-gingerol on hair growth in C57BL/6 rats was measured through topical application of 6-gingerol on the dorsal skin of each animal.

RESULTSThe length of hair shaft in 20 microg/ml 6-Gingerol group (0.50 +/- 0.08 mm) is less than 0 microg/ml (0.66 +/- 0.19) mm and 10 microg/ml (0.64 +/- 0.03) mm 6-Gingerol group (P < 0.05). In cell culture, compared to 0 microg/ml and 5 microg/ml 6-Gingerol, 10 microg/ml 6-Gingerol can significantly inhibited the proliferation of DPCs (P < 0.05). Along with the growth inhibition of DPCs by 6-gingerol, the Bax/Bcl-2 ratio increased obviously. In vivo study, the hair length and density decreased a lot after using 1 mg/ml 6-gingerol.

CONCLUSIONS6-Gingerol can suppress human hair shaft elongation because it has pro-apoptotic effects on DPCs via increasing Bax/Bcl-2 ratio. It might inhibit hair growth by prolonging the telogen stage in vivo.

Animals ; Catechols ; pharmacology ; Cell Culture Techniques ; Cells, Cultured ; Fatty Alcohols ; pharmacology ; Hair ; drug effects ; growth & development ; Hair Follicle ; drug effects ; growth & development ; Humans ; Mice ; Mice, Inbred C57BL ; Plant Extracts ; pharmacology ; Rats ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo examine the spatial distribution of dead and vital bacteria in the early formation of native dental biofilm.

METHODSAn experimental dental biofilm model in the oral cavity was established by enamel slabs and the spatial distribution of dead and vital bacteria in the early colonization of native dental biofilm on the enamel surface was observed by in situ real-time and dynamic observations and optical sections utilizing confocal laser scanning microscope (CLSM) and live and dead bacterial fluorescence staining technique.

RESULTSAt the initial stage of dental biofilm formation, the structure was sparse and the percentage of dead cells reached 70% - 80% at the inner layers. In the middle layers the structure became denser than in the inner layers, which was mainly composed of vital cells (40% - 70%), and void-like structures were surrounded by vital bacteria. In the outer layers, the structure was sparse and vital cells occupied 20% - 40%.

CONCLUSIONSNative dental biofilms showed an uneven spatial distribution of vital and dead microorganisms. The percentage of vital microorganisms was lower adjacent to the enamel surface, increased in the z-axis towards the central parts, and decreased again towards the outer layers. The dead bacteria is an integral component in the early formation of native dental biofilm. Bacteria in the biofilms increased with time forming abundant channels.

Adult ; Biofilms ; Dental Plaque ; microbiology ; Humans ; Microscopy, Confocal ; Staining and Labeling ; Young Adult

Studies have found that metformin is not only the preferred drug for lowering blood sugar, but also shows lipid-lowering and weight-loss effects. The purpose of this study was to use a hyperlipidemia hamster model to investigate the lipid-lowering effect of metformin and its effect on important metabolic pathways in lipid metabolism disorders. Fifty golden hamsters were divided into a control group, a model group, metformin high- and low-dose groups, and a simvastatin group. A high-fat diet was fed for 1 week to create the model, and then drug was administered for 11 weeks with the high-fat diet. Serum was taken for measurement of blood lipid and blood glucose at 2, 6, and 9 weeks after administration, and at weeks 3, 5, and 9 feces and urine were collected for ¹H NMR metabolomics tests. After 11 weeks of intravenous injection of [U-¹³C₆] glucose, serum was collected for a ¹³C NMR metabolic flux test. The results showed that the administration of metformin can significantly reduce blood lipids and glucose levels and can significantly affect metabolic pathways such as sugar metabolism, lipid metabolism, ketone metabolism, amino acid metabolism, and intestinal flora metabolism. The results of the metabolic flux analysis showed that the high-fat diet reduced the metabolism of tricarboxylic acids by 37.48%. After administration of low and high doses of metformin the metabolism of tricarboxylic acid increased by 98.14% and 143.10%, respectively. After administration of simvastatin tricarboxylic acid metabolism increased by 33.18%. The results indicate that metformin has a significant effect on promoting energy metabolism. This study used a combination of metabolomics and metabolic flow to explore the effect of metformin on lipid metabolism disorders and quantifies changes in the key pathway of energy metabolism-the tricarboxylic acid cycle. This study provides useful information for the study of the efficacy and mechanism of metformin, as well as a practical technical method for the screening of lipid-lowering drugs based on a hamster model.