OBJECTIVETo establish a method for isolating and culturing mouse dental follicle cells and to study the phenotype characteristics of dental follicle cells.

METHODSMandibular first molars from 9 day old Balb/c mice were digested with 1% trypsin, subsequently, and the dental follicle was enucleated from the tooth germ and cultured. The shape and ultrastructural appearance of dental follicle cells were observed under phase-contrast microscope and scanning electron microscope (SEM). Immunocytochemistry was used to detect the expression of alkaline phosphatase (ALP), bone sialoprotein (BSP) and osteopontin (OPN).

RESULTSThree types of cells were isolated: some were cuboidal/polygonal; some were elongated, spindle-shaped, fibroblast-like cells; and a minor, third cell type was very thin and elongated. The cytoplasm of the first two cell types was filled with abundant granules. The cells were pleomorphism under SEM and had many filipodia and microvilli. According to whether there were filaments overlying the surface, the cells could be divided into two subtypes. Some but not all follicle cells expressed ALP, BSP and OPN.

CONCLUSIONThe cultured dental follicle cells consisted of several cell phenotypes and had the potential of differentiating into cementoblasts, periodontal ligament fibroblasts and osteoblasts.

Alkaline Phosphatase ; Animals ; Cell Culture Techniques ; Cell Line ; Cells, Cultured ; Dental Cementum ; Dental Sac ; Fibroblasts ; Integrin-Binding Sialoprotein ; Mice ; Mice, Inbred BALB C ; Molar ; Osteoblasts ; Osteopontin ; Periodontal Ligament ; Phenotype ; Tooth Germ

Objective To establish qualitative and quantitative analysis methods for identification and determination of multi-constituent of Chanfukang granules (CFKG) using high performance liquid chromatography-time-of-flight mass spectrometry (HPLC-TOF-MS) and high performance liquid chromatography-triple quadrupole mass spectrometry (HPLC-MS/MS),and a preliminary pharmacokinetic study was carried out.Methods An Agilent ZORBAX Eclipse Plus C1s column (100 mm × 3.0 mm,1.8 μm) and a Waters XBridge(R) BEH C18 column (150 mm × 4.6 mm,2.5 μm) were used with 0.1％ formic acid aqueous solution-acetonitrile as mobile phases by gradient elution.The compounds were detected by electrospray ion source in both positive and negative mode with multiple reaction monitoring (MRM) mode.SD rats were ig with CFKG (0.5,5 g/kg).Blood samples of 0.2 mL were taken from jugular vein at different time points and plasma components and contents were determined by HPLC-MS/MS.Results Twenty-four constituents were identified by HPLC-TOF-MS qualitative analysis and thirteen constituents were quantitatively detected by HPLC-MS/MS.The established HPLC-MS/MS method had a good separation,and all the legal verification met the requirements.The content of stachydrine,Leonurine,and astragaloside Ⅳ were high,which may be the main active ingredient The pharmacokinetic results showed that,only when the dose was 5 mg/kg,stachydrine and Leonurine can detected.Elimination of stachydrine was slow and elimination of Leonurine was fast.Conclusion A rapid and efficient method for studying the chemical constituents was established,which could provide reference for the quality control of Chanfukang granules.

OBJECTIVETo compare the hybrid between species of Dendrobium huoshanense and its parents on growing, physiologic indexes and content of medicinal components, and provide theoretical basis for species quality improvement.

METHODThe chlorophyll content, the photosynthesis rate, the polysaccharides content and the alkaloids content were measured by anhydrous ethanol method, Cl-310 photosynthesis determination system, colorimetry of concentrated sulphuric acid-phenol and acid dyes colorimetry respectively.

RESULTThe growth of hybrid was close to D. moniliforme, and apparently higher than D. huoshanense. The chlorophyll content and the photosynthesis rate of one-year-hybrid were markedly higher than its parents. The content of polysaccharides and alkaloids in two-year-stem and three-year-stem of hybrid were close to that of D. huoshanense.

CONCLUSIONThe hybrid integrates superiority of parents on growth and accumulation of medicinal components opens vast vistas for development and utilization.

Alkaloids ; analysis ; Chlorophyll ; analysis ; Dendrobium ; chemistry ; classification ; genetics ; growth & development ; Hybridization, Genetic ; Photosynthesis ; Plants, Medicinal ; chemistry ; classification ; genetics ; growth & development ; Polysaccharides ; analysis

The purpose of this study was to investigate the repair of the osteoarthritis(OA)-induced cartilage injury by transfecting the new TGF-β3 fusion protein (LAP-MMP-mTGF-β3) with targeted therapy function into the bone marrow-derived mesenchymal stem cells (MSCs) in rats. The recombinant of pIRES-EGFP-MMP was constructed by combination of DNA encoding MMP enzyme cutting site and eukaryotic expression vector pIRES-EGFP. LAP and mTGF-β3 fragments were obtained from rat embryos by RT-PCR and inserted into the upstream and downstream of MMP from pIRES-EGFP-MMP respectively, so as to construct the recombinant plasmid of pIRES-EGFP-LAP-MMP-mTGF-β3. pIRES-EGFP-LAP-MMP-mTGF-β3 was transfected into rat MSCs. The genetically modified MSCs were cultured in medium with MMP-1 or not. The transfected MSCs were transplanted in the rat OA models. The OA animal models were surgically induced by anterior cruciate ligament transaction (ACLT). The pathological changes were observed under a microscope by HE staining, Alcian blue, Safranin-fast Green and graded by Mankin's scale. pIRES-EGFP-LAP-MMP-mTGF-β3 was successfully constructed by means of enzyme cutting and sequencing, and the mTGF-β3 fusion protein (39 kD) was certified by Western blotting. Those genetically modified MSCs could differentiate into chondrocytes induced by MMP and secrete the relevant-matrix. The transfected MSCs could promote chondrogenesis and matrix production in rat OA models in vivo. It was concluded that a new fusion protein LAP-MMP-mTGF-β3 was constructed successfully by gene engineering, and could be used to repair the OA-induced cartilage injury.

Child ; Electrocardiography ; Humans ; Male ; Ventricular Fibrillation ; physiopathology

Objective To investigate the prevalence of endemic fluorosis and the progress acchieved through control measures in Taian County of Gansu Province so as to provide basis and technique support for endemic fluorosis control. Methods The survey was carried out according to the National Surveillance Scheme of Endemic Fluorosis. Fulfillment of control measures and the quality of water improving projects were surveyed. Samples of household water and urine in children aged 8-12 years were collected and fluoride content was detected by iron selective electrode method. Children's dental fluorosis were examined with Dean method. Results In Qinan County, water was improved in a rate of 100%. In water-improving and defluoridation projects investigated, 81.15% (99/122) projects worked well, 18.85% (23/122) projects were closed or abandoned. The number of water improvement projects monitored were 24,81,9,8,respectively in the year through 2004 to 2007, revealing 13 projects having water fluorine content(>1.0-2.0 mg/L) in 2004, 15 and 5 projects having water fluorine content higher that 1.0-2.0 and 2.0-4.0 mg/L respectively in 2005, 2 projects having water fluorine content(>1.0-2.0 mg/L) in 2006, and no projects above 4.0 mg/L. In Anfu Village, the fluorine content of source and drinking water were all below 1.0 mg/L in 2006 and 2007. The urinary fluoride content of 8-12 years old children was equal and higher than 1.5 mg/L, dental fluorosis rate was 34.47% (354/1027), and dental fluorosis index was 0.65 in 2004-2007. Conclusions Projects running out of status and excessive levels of water fluorine are frequently seen, dental fluorosis is not controlled in Taian County of Gansu Province

Objective To evaluate the reliability and validity of the application of condensed Chinese version of the MOS 36-item short form health survey (SF-36) in assessment of quality of life among adult patients with Kashin-Beck disease,and to provide a scientific basis in rehabilitation of the patients.Methods Four hundred and twenty seven eases of adult patients with Kashin-Back disease and 419 healthy individuals randomly selected in Kashin-Beck disease endemic areas in 8 counties of Gansu province were surveyed with the SF-36.The reliability of the SF-36 was assessed by split-half reliability and Cronbach's α coefficient and the validity through principal component factor analysis and correlation analysis,etc.The dimension scores of different people were obtained by analysis of variance and univariate t-test.Results The split-half reliability of all the 8 dimensions was greater than 0.6 and the Cronbach's α coefficient was greater than 0.8; the pearson correlate coefficients of all the items to their dimensions were greater than 0.391.SF-36 contained 8 domains and 2 summary scales in the factor analysis.The score differences of quality of life in different ages of the patients,different stages of the disease were statistically significant (all P ＜ 0.05).Conclusion The SF-36 is practical in studying the quality of life among adult patients with Kashin-Beck Disease.

OBJECTIVETo compare the results of cytogenetic and IPSS grouping of primary myelodysplastic syndromes (pMDS) patients classified by FAB- or WHO classification.

METHODSTwo hundred and thirty seven cases of pMDS who were previously classified according to FAB criteria were reclassified with WHO classification. A comparison was made between the results of the two classifications.

RESULTSFor the detection rates of cytogenetic abnormality and its risks group, there was no difference among the FAB subgroups but the detection rate was different between the WHO refractory cytopenia with multilineage dysplasia (RCMD) and RA subgroups (74.4% and 42.5%, respectively) (P < 0.001). The percentage of good karyotype abnormalities in RA (65.0%) was higher than that in RCMD (24.4%) (P < 0.001), and the percentages of intermediate and poor karyotype abnormalities in RCMD (48.9% and 26.7%, respectively) were higher than that in RA (27.5% and 7.5%, respectively) (P < 0.05). There was a good correlation between the subgroups and IPSS risk groups for both the WHO classification and the FAB classification, but the WHO classification further reflected the differences between RCMD and RA and RAEB-I and RAEB-II subgroups. The percentage of low-risk group in RCMD (1.1%) was lower than that in RA (10.0%) (P < 0.05), and the percentage of high-risk group in RAEB-II (30.5%) was higher than that in RAEB-I(0) (P < 0.001).

CONCLUSIONFor the correlation between subgroups and cytogenetic abnormalities and IPSS risk groups, the WHO-classification is better than the FAB-classification.

Adolescent ; Adult ; Aged ; Bone Marrow ; pathology ; Child ; Child, Preschool ; Female ; Humans ; Karyotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; classification ; genetics ; pathology ; Prognosis ; Severity of Illness Index ; World Health Organization

OBJECTIVETo investigate the expression of epidermal growth factor receptor (EGFR) during the mineralization of human periodontal ligament cells (hPDLC) in vitro.

METHODSStudies using specific antibodies to immunolocalize EGFR in the mineral differentiating hPDLC were undertaken to investigate the different expression during the inducing process. In situ hybridization and RT-PCR technique were used to investigate the transcripts encoding the protein of EGFR.

RESULTSThe results showed that immunocytochemical labeling gradually decreased following the elong of the induce time, downing to nearly negative at the 4th week and the signal of EGFR transcripts was weaker in the induced hPDLC than that in uninduced.

CONCLUSIONEGFR has a negative regulation function during the mineralization of hPDLC.

Cells, Cultured ; Humans ; In Situ Hybridization ; In Vitro Techniques ; Periodontal Ligament ; Receptor, Epidermal Growth Factor