[ ABSTRACT] AIM:To investigate the distribution of mast cells ( MCs) and the expression of transforming growth factor-β(TGF-β) on tryptase positive MCs in different types of human periapical diseases.METHODS:Total 78 cases of specimens were involved in this study, including healthy control, periapical cyst and periapical granuloma.The tissue sam-ples were fixed in 10% formalin for at least 48 h, stained with hematoxylin and eosin for histopathological examination, stained with toluidine blue staining for identifying MCs and MCs degranulation, and stained with double immunofluores-cence for identification of tryptase-TGF-βdouble positive MCs.RESULTS:The density of tryptase-TGF-βdouble positive MCs in the periapical lesions was significantly higher than that in the healthy controls ( P<0.01) .The number of TGF-βpositive MCs in the periapical cyst was significantly higher than that in the periapical granuloma ( P<0.01 ) .Compared with toluidine blue staining, the number of MCs with double immunofluorescence staining significantly increased ( P <0.01).CONCLUSION:The TGF-βpositive MCs may play an important role in the pathogenesis of human chronic peria-pical diseases, particularly in the formation of fibrous tissue in periapical cyst.Double immunofluorescence staining is more sensitive than the traditional toluidine blue staining for identifying MCs.

Objective To study the optimum determination conditions for lymphocytic proliferation by CCK-8 method in mice.Methods To study the different influence factors of spleen cell proliferation experiment stimulated by mitogen concanavalin A (ConA) or lipopolysaccharide (LPS),including cell preparation method,lymphocytic density,FBS and stimulating agent concentration in culture medium,and stimulating immediately or 24 h after preparing cell,with cross design or two factor completely randomized design.Results Spleen lymphocytic proliferation rate of preparation method by light suppression was higher than that of the light grind.The appropriate concentration of spleen cells was 5 × 106/mL.The proliferation rate has no significant difference after being stimulated for 48 or 72 h by ConA (2,5,or 1 0 μg/mL) or LPS (10,20,or 50 μg/mL) under 10％,15％,or 20％ FBS concentration in culture medium.The proliferation rate of stimulating immediately after preparing cell was higher than that of 24 h after preparing cell.Conclusion The optimum conditions of Balb/C mouse spleen cell proliferation assay stimulated by ConA and LPS are as follows:preparation of spleen cells with light pressure,spleen cell concentration of 5 × 106/mL,direct stimulation with 2-10 μg/mL ConA or 10-50 μg/mL LPS in the day of preparation.

Objective To evaluate the role of large conductance calcium-activated potassium (BKCa) channels in vascular hyporesponsiveness in rats with obstructive jaundice.Methods Eighteen male Sprague-Dawley rats,weighing 180-200 g,were randomly divided into 3 groups (n =6 each) using a random number table:control group (group C),sham operation group (group S),and bile duct ligation group (group BDL).Obstructive jaundice was produced by common bile duct ligation.At 7 days after surgery,blood samples were collected for determination of the levels of serum total bilirubin (TBL),direct bilirubin (DBL),indirect bilirubin (IBL),alanine aminotransferase (ALT),and aspartate aminotransferase (AST).Thoracic aortic rings were prepared,and the endothelium was removed.The aortic rings were sequentially perfused with different concentrations of norepinephrine (NE) and sodium nitroprusside (SNP),and the maximum amplitude of contraction and dilatation of aortic rings was recorded.The aortic rings were then perfused with BKCa channel blocker Chtx with the final concentration of 10 7 mol/L,followed by perfusion with different concentrations of NE and SNP again,and the maximum amplitude of contraction and dilatation of aortic rings was recorded under each concentration.The percentage of maximum contraction and dilatation (maximum amplitude after Chtx administration÷maximum amplitude before Chtx administration× 100％) was calculated.Results Compared with C and S groups,the levels of TBL,DBL,IBL,ALT and AST in serum were significantly increased,the maximum amplitude of NE-induced contraction of aortic rings was decreased,and the percentage of the maximum NE-induced dilatation of aortic rings was increased,the maximum amplitude of SNP-induced contraction of aortic rings was increased,and the percentage of the maximum SNP-induced dilatation of aortic rings was decreased in group BDL.Conclusion Excessivc opening of BKCa channels may be involved in the mechanism of vascular hyporesponsiveness in rats with obstructive jaundice.

Objective: To investigate the application of one stage otoptasty for congenital atresia of the external acoustic canal and malformations of the middle ear and the auricle. Method: patients with the ear malformations were given surgical reconstruction of one stage otoplasty. The auricle was reconstructed with the rib which was encapsuled with the superthin temporal flap. According to the malformations of the middle ear in patients, Ⅰ style tympanoplasty and Ⅲ style tympanoplasty were carried out respectively. All patients were performed myringoplasty with temporal fascia and reconstructed the external acoustic canal with full thickness skin-grafting. Result:A long term follow-up (4～6 years)demonstrated that 11 ears were survival of which 8 ears figuration were ideal. The hearing improvement was observed in all patients. Conclusion:one stage otoplasty is effective for treatment of the congenital malformations of the external and middle ear.

OBJECTIVETo investigate the distribution in rat after oral administration of Smilax china extract.

METHODOxyresveratrol and resveratrol were quantitatively determined by HPLC/DAD method in rat tissues including heart, liver, spleen, lung and kidney after oral administration of S. china extract. The separation was performed on a Zorbax SB-C18 column (4.6 mm x 250 mm, 5 microm) with acetonitrile-water using linear gradient elution and UV detection at wavelength of 320 nm. The flow rate was 1.0 mL x min(-1) and the column was kept at 40 degrees C.

RESULTThe HPLC/DAD method for the simultaneous determination of oxyresveratrol and resveratrol in rat tissues was developed.

CONCLUSIONThe method has been successfully applied for the distribution study of two active constituents in rat through oral administration of S. china extract.

Administration, Oral ; Animals ; Chromatography, High Pressure Liquid ; methods ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Female ; Plant Extracts ; administration & dosage ; analysis ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Smilax ; chemistry ; Stilbenes ; administration & dosage ; analysis ; pharmacokinetics

To optimize program of bovine somatic nuclear transfer, we used two different enucleation procedures (by Spindle-view system & Hoechst 33342 staining), two different procedures to introduce donor nuclei (by ooplasm microinjection & electrofusion), and three different group electrofusion parameters (group 1: 1.9 kV/cm, 10 micros, two; group 2: 1.5 kV/cm, 25 micros, two; group 3: 0.6 kV/cm, 100 micros, one) to reconstruct bovine cloned embryos. The cleavation rates and blastocyst development rates of cloned embryos were used to assess the efficiency of different operational procedure. Finally, the best combination of operational procedure, that the spindle-viewer system was used for oocytes enucleating, and donor cell was electrofused into ooplasm by electrical pulse (1.9 kV/cm, 10 micros, two) to reconstruct bovine cloned embryos. Then the excellent blastocysts were transferred to fosters for producing cloned cattle 80 high-quality cloned blastocysts were transferred into 33 fosters, two cloned calves were produced. According to the results, the optimized program could be used to produce cloned cattle.
Animals ; Cattle ; Cell Nucleus ; physiology ; Cloning, Organism ; veterinary ; Embryo Transfer ; methods ; Embryo, Mammalian ; cytology ; physiology ; Female ; Microinjections ; Nuclear Transfer Techniques ; veterinary ; Oocytes ; cytology ; physiology