[Graded ethanol precipitation method on physicochemical properties and antioxidant activities of polysaccharides extracted from Astragalus Radix].

Astragalus polysaccharide has been widely used in food and medicinal industry owing to its health-promoting properties. In order to characterize better the relationship among molecular weight, structure-activity and activities, a simple method was used different concentration of ethanol including 30% (PW30), 50% (PW50), 70% (PW70), 75% (PW75), 80% (PW80) and 90% (PW90) to precipitate Astragalus polysaccharides into different molecular weight. As a result, PW90 showed smooth surface and the strongest antioxidant activity among these six fractions (P < 0.05). In conclusion, graded ethanol precipitation was a simple method to separate Astragalus polysaccharides into different molecular weight with different antioxidant activity fractions.

[Dynamic monitoring risk of anti-hepatoma new drug development].

Risk monitoring of new Chinese patent anti-hepatoma drugs is tracking recognized risks and residual risks, identifying emerging risk and ensure the implementation of the plan, estimating the process of reducing effectiveness. The paper is mainly through understanding the status of Chinese patent anti-hepatoma drugs, the content, characteristic and analysis method of dynamic risk monitoring, and then select the risk control indicators, collect risk information. Finally, puts forward the thought of anti-hepatoma drugs listed evaluation in our country, and try to establish the model of dynamic risk management of anti-hepatoma drugs.

[Regularity of drugs compatibility of anti-hepatoma traditional Chinese medicine ancient prescriptions and risk evaluation of anti-hepatoma new drug research and development].

Traditional Chinese ancient prescriptions have been used for treatment of liver cancer for a long history and the scientific and rational compatibility is a great wealth for modern research and development (R&D) of new drugs. The research and development of new drugs are often accompanied with a large investment, a long cycle and a high risk, especially for the anti-tumor drugs R&D which are facing more risks and lower successful rate. In this research, the regularity of compatibility of drugs was analyzed from 124 anti-hepatoma ancient prescriptions by computer program. The results can offer help to the R&D of anti-hepatoma new drugs and reduce the risk of drug screening. In addition, we surveyed 22 companies in this field from six provinces such as Beijing, Shanghai, Tianjin and so on and obtained 240 risk assessment questionaires. Then we used qualitative analysis method to interpret the greatest impacts for the risks in the process of R&D, production and sales of anti-hepatoma new drugs. The study provides a basis for anti-liver cancer drugs R&D researchers, who can take effective measures to reduce the R&D risks and improve successful rate.

[Expression of Osterix mRNA and protein induced by recombinant human bone morphogenetic protein-2 in human periodontal ligament cells].

OBJECTIVE: To detect the changes of Osterix (Osx) mRNA and protein expression in human periodontal ligament cells (HPDLCs) induced by recombinant human bone morphogenetic protein-2 (rhBMP-2), and examine the role of BMP-2 and Osx during the osteogenic differentiation of HPDLCs. METHODS: HPDLCs were isolated and cultured in vitro with explant method. Cells at passage 3 were cultured in DMEM with rhBMP-2 at different concentrations (50, 100, 150, 200, 250, 300, 400, and 600 µg/L) for different times (2, 3, 5, 7, 10, 14 and 21 days). Then the expressions of Osx mRNA and protein were measured by real-time reverse transcription (RT)-PCR and Western blotting respectively. Cells were treated with 10 µmol/L SB203580 (p38 inhibitor) to inhibit p38 phosphorylation induced by rhBMP-2. The mineralization nodules formation and the expressions of phosphorylated p38 and Osx mRNA were detected respectively. RESULTS: During the culture of rhBMP-2, the expression of Osx mRNA significantly increased. Initially Osx protein had a low expression and then increased in a time-dependent manner followed by the production of bone-like nodules in HPDLCs. Under the effect of SB203580, the up-regulation of phosphorylated p38 expression induced by rhBMP-2 was significantly inhibited as well as the expression of Osx (Osx mRNA expression: 0.378 ± 0.034 vs 0.134 ± 0.027, Osx protein expression: 0.353 ± 0.024 vs 0.155 ± 0.031, both P < 0.01). Meanwhile the mineralization nodules formed by HPDLCs induced by rhBMP-2 were fewer and delayed. CONCLUSIONS: BMP-2 has a significant positive regulatory role on the expression of Osx in HPDLCs. And p38 pathway is an important link of this regulatory process. Thus, as an important signaling pathway in osteogenic differentiation of HPDLCs, BMP-2/p38/Osx may be involved in periodontal tissue remodeling.

[Comparing the effects of fast and slow expansion on nasal cavity and maxilla structure].

OBJECTIVE: This study aims to compare the effects of fast and slow expansion on nasal cavity structure. METHODS: A total of 40 patients were selected and randomly divided into two groups. Cone-beam computer tomography (CBCT) was obtained before and after surgery and used for comparing the changes in nasal structure before and after treatment. RESULTS: Fast expansion had resulted in greater changes in the basilar and nasal bone arch extension structures than slow expansion. No significant difference at maxillary width and nasal parenchyma. CONCLUSIONS: Rapid expansion therapy has more beneficial effects on nasal function.

[Detection and significance of phosphatidylinositol 3-kinase in adipose tissue of polycystic ovary syndrome patients with insulin resistance].

OBJECTIVE: To investigate the expression of phosphatidylinositol 3-kinase (PI-3K) in adipose tissue of polycystic ovary syndrome patients (PCOS), and explore molecular mechanisms of insulin resistance (IR) in PCOS. METHODS: Samples from patients with PCOS with IR (n = 19), PCOS without IR (n = 10) and controls (n = 15) were collected. Serum fasting insulin (FIN) and fasting plasma glucose (FPG) were measured. Insulin resistance index was calculated using homeostasis model assessment (HOMA) to analyze the relationship between these markers and IR. Western blot technique was used to detect the PI-3K p85 subunit. Gene expression of PI-3K p85 subunit was detected by reverse transcription polymerase chain reaction (RT-PCR) method. Kinase activity was detected by immunoprecipitation, thin-layer chromatography and gamma scintillation counting. RESULTS: (1) The levels of FIN [(25.2 +/- 3.8) mU/L] and HOMA-IR (1.6 +/- 0.3) in PCOS with IR were significantly higher than those in PCOS without IR [(13.4 +/- 3.8) mU/L, 0.9 +/- 0.3] and controls [(9.5 +/- 2.6) mU/L, 0.5 +/- 0.3; all P < 0.05). (2) There was no significant difference in the protein (0.65 +/- 0.10) and gene expression (0.92 +/- 0.12) of PI-3K p85 subunit in PCOS with IR compared with PCOS without IR (0.72 +/- 0.10, 1.01 +/- 0.10) and control groups (0.73 +/- 0.14, 1.00 +/- 0.12; P > 0.05). (3) PI-3K activity in PCOS with IR (81%) and PCOS without IR (89%) was significantly decreased (P < 0.01, P < 0.05) and negatively correlated with HOMA-IR (r = -0.69, P < 0.01; r = -0.62, P < 0.05). CONCLUSIONS: No significant difference in the protein and gene expression of PI-3K p85 subunit in PCOS with IR is found. The decreased PI-3K activity may lead to IR of PCOS.

[Tyrosine phosphorylation and protein expression of insulin receptor substrate-2 in the adipose tissue from patients with polycystic ovary syndrome].

OBJECTIVE: To explore molecular mechanisms of insulin resistance of polycystic ovary syndrome (PCOS) by determining the tyrosine phosphorylation and protein expression of insulin receptor substrate-2 (IRS-2) in adipose tissue from patients with PCOS. METHODS: Serum and subcutaneous adipose tissue samples from patients with PCOS with insulin resistance (n = 19), PCOS without insulin resistance (n = 17) and controls (n = 20) were collected. The expression of IRS-2 in adipose tissue was assessed by Western blot. Immunohistochemistry was used to detect the distribution of IRS-2 in adipose tissues of all patients. The tyrosine phosphorylation of IRS-2 was measured by immunoprecipitation and enhanced chemiluminescent immunoblotting technique. RESULTS: (1) There was no significant difference of the protein expression of IRS-2 in PCOS with insulin resistance 1.15 +/- 0.26 compared to those in PCOS without insulin resistance 1.13 +/- 0.26 and control group 1.00 +/- 0.25 (P > 0.05); (2) The tyrosine phosphorylation of IRS-2 was significantly decreased in PCOS with insulin resistance 0.77 +/- 0.16 compared to that of PCOS without insulin resistance 0.91 +/- 0.25 and control groups 1.00 +/- 0.12 (P < 0.05). There was no significant difference between PCOS without insulin resistance and control groups (P > 0.05). CONCLUSIONS: The decrease of tyrosine phosphorylation of IRS-2 in PCOS patients, which induces impairment of the insulin signal pathway, may be one of the mechanisms leading to insulin resistance.

[Cone-beam CT study for the oropharyngeal airway volume and hyoid position of adults Class III skeletal malocclusion].

PURPOSE: Using cone-beam CT (CBCT) and Invivo5 software to measure the oropharyngeal airway volume and hyoid position of adults Class III skeletal malocclusion and analyze the influence of the maxillary or mandibular position, in order to evaluate oropharynx airway volume and hyoid position ,and provide guidance for reasonable and effective treatment. METHODS: One hundred and eighty cases were randomly selected from CBCT data and divided into three groups. Invivo5 software was used to rebuild 3D image data and establish a 3D coordinate system, then to measure OPV and hyoid position. LSD t test was performed between each group with SPSS 19.0 software package. RESULTS: OPV for mandibular protrusion of Class III was significantly higher than maxillary retrusion of Class III and Class I (P<0.05), whereas no difference existed between Class I and maxillary retrusion of Class III; H-VPS had significant difference as follows: mandibular retrusion of Class III, maxillary retrusion of Class III,and Class I (P<0.05); H-PS in mandibular protrusion of Class III was significantly higher than maxillary retrusion of Class III and Class I (P<0.05), whereas no difference existed between Class I and maxillary retrusion of Class III. CONCLUSIONS: The three-dimensional features of airway were different as adults have different types of skeletal malocclusion. The mandibular sagittal position and length had more significant influences on pharyngeal airway volume, shape, direction, and hyoid sagittal position compared with the maxilla. CBCT and Invivo5 are practical tools for evaluation of airway, which can provide clinical evaluation of the three-dimensional features of airway and hyoid position of different skeletal malocclusion, and provide guidance in the formulation of reasonable and effective treatment plan.

[Tyrosine phosphorylation and protein expression of insulin receptor substrate-1 in the patients with polycystic ovary syndrome].

OBJECTIVE: To investigate the tyrosine phosphorylation and protein expression of insulin receptor substrate-1 in adipose tissue from patients with polycystic ovary syndrome, and explore molecular mechanisms of insulin resistance of PCOS. METHODS: Samples from patients with PCOS with insulin resistance (group A, n = 19), PCOS without insulin resistance (group B, n = 10) and controls group (n = 15) were collected. Serum luteinizing hormone (LH), follicle stimulating hormone (FSH) and testosterone (T) were measured by chemiluminescence assay. Fasting insulin (FIN) was measured by radioimmunoassay. Fasting plasma glucose (FPG) was measured by oxidase assay. Insulin resistance index was calculated using homeostasis model assessment (HOMA) to analyze the relationship between these markers and insulin resistance. The amount of insulin receptor substrate-1 in adipose tissue was assessed by western blot. The tyrosine phosphorylation of IRS-1 was measured by immunoprecipitation. RESULTS: (1) The levels of serum LH (15.8 +/- 2.8) U/L, LH/FSH 2.8 +/- 0.6, T (4.3 +/- 0.9) nmol/L, FIN (25.2 +/- 3.8) mU/L and HOMA IR (1.56 +/- 0.25) in group A were significantly higher than those of group B (13.9 +/- 1.9) U/L, 2.3 +/- 0.4, (3.6 +/- 0.4) nmol/L, (13.4 +/- 3.8) mU/L, 0.87 +/- 0.28 and control group (7.3 +/- 2.1) U/L, 1.3 +/- 0.3, (0.9 +/- 0.2) nmol/L, (9.5 +/- 2.6) mU/L, 0.50 +/- 0.30 (all P < 0.05); (2) The protein expression and tyrosine phosphorylation of IRS-1 in group A (690 +/- 19 and 528 +/- 72 respectively) were significantly lower than those in group B (892 +/- 31, 801 +/- 64) and control group (988 +/- 29, 1139 +/- 124) (P < 0.05, and P < 0.01 respectively). (3) Insulin resistance index in group A and group B were negatively related with protein expression and tyrosine phosphorylation (r = -0.52, P < 0.05; r = -0.61, P < 0.01 and r = -0.60, P < 0.05; r = -0.63, P < 0.05). CONCLUSIONS: The signal transduction malfunction because of protein expression and tyrosine phosphorylation changes of IRS-1 in adipose tissue from polycystic ovary syndrome patients may be one of the mechanisms leading to insulin resistance.

[Relationship between abnormal swallowing and mouth breathing].

OBJECTIVE: To investigate the relationship between abnormal swallowing and mouth breathing. METHODS: Thirty-eight patients with abnormal swallowing and 38 patients with normal swallowing were selected. All patients presented with no airway constriction. The age range of the patients was 11-14 years old. The number of patients with mouth breathing was calculated. Statistical analysis (χ(2) test) was performed. RESULTS: The number of patients with mouth breathing in the abnormal swallowing group (17, 45%) was significantly higher than that in the normal swallowing group (5, 13%) (χ(2) = 9.212, P = 0.002). CONCLUSIONS: Abnormal swallowing was related to mouth breathing.

Novel cartilage-derived biomimetic scaffold for human nucleus pulposus regeneration: a promising therapeutic strategy for symptomatic degenerative disc diseases.

Because current therapies have not always been successful and effective, the possibility of regenerating the nucleus pulposus (NP) through a tissue-engineered construct offers a novel therapeutic possibility for symptomatic degenerative disc diseases (DDDs). However, more research is necessary to identify the optimal scaffold, cell type and mixture of signal factors needed for NP regeneration. Numerous possible scaffolds for NP regeneration have been investigated; they have many shortcomings in common. Various biological scaffolds derived from decellularized tissue and organs have been successfully used in tissue engineering and received approval for use in humans. Regretfully, harvesting of human NP is difficult and only small amounts can be obtained. The macromolecules of cartilage, which include collagen and proteoglycan aggrecan, are similar to those of the extracellular matrix of immature NP. Recent studies have shown that adipose-derived stem cells (ADSC) can be induced to develop NP-like phenotypes when stimulated by appropriate signals. We thus reasonably postulated that an ideal NP scaffold for tissue engineering could be fabricated from decellularized cartilage matrix (DCM). Furthermore, a combination of ADSCs and DCM-derived biomimetic scaffolds would be advantageous in NP tissue engineering and, in the long run, could become an effective treatment option for symptomatic DDD.

[Expression of Osterix mRNA and protein levels in cultured human periodontal ligament cells after mechanical stimulation].

OBJECTIVE: To examine the expression of Osterix (Osx) mRNA and protein after application of mechanical force on human periodontal ligament cells (HPDLCs), and to investigate the role of Osx in orthodontic alveolar bone remodeling. METHODS: HPDLCs were isolated and cultured in vitro with explant method. Approximately 2.5 x 10(5) cells were seeded onto six-well cell culture plates and then were exposed to centrifugal force for 1, 2, 4, 6, 8 or 12 h at 631 r x min(-1). The expression of Osx mRNA and protein was measured by reverse transcription-polymerase chain raction (RT-PCR) and Western blot respectively. Immunofluorescence assay was used to detect the expression and subcellular At the initial time point, Osx mRNA had a weak exlocalization of Osx protein by green fluorescence. RESULTS: pression and protein was not detected. Under the mechanical stimulation, both mRNA and protein levels of Osx were upregulated in a time-dependent manner. Furthermore, Osx protein was translocated gradually from the cytosol into the cell nuclei. CONCLUSION: The expression and activation of Osx were enhanced by mechanical stress in HPDLCs, which indicates that Osx may play an important role in HPDLCs osteogenic differentiation and periodontal tissue remodeling induced by mechanical stress.