0.05). Conclusion Aspirin-vitamin C dispersible tablets is effective and safe for treating patients with URI.

OBJECTIVETo establish a simple and highly effective isolation and culture system of mouse spermatogonial stem cells(SSCs)and detect the expression of stem cell-related markers in the isolated cells.

METHODSThe structures of seminiferous tubules of neonatal(6-8 days of age)and adult(26-28 weeks)DBA/2 mice were compared using histochemical examination. Testes of neonatal mice were selected for preparing primary cells. The digestive efficiency of different enzymes was compared. SSCs were isolated according to the different binding abilities of testicle somatic cells and SSCs to gelatin matrix. The effects of different base culture media such as StemPro34 and α-MEM,gelatin,and serum on the SSCs binding activity and growth were studied. The cell morphology was observed during the culture process. Immunofluorescence was used to detect the expression of SSCs and cancer stem cells(CSCs)-related markers in SSCs.

RESULTSThe content of SSCs in the testes of neonatal mice was relatively higher than that in adult mice. Trypsin showed the highest digestive efficiency. In StemPro34 supplemented with 1% fetal bovine serum and on the gelatin matrix,testicular somatic cells could bind with the plate efficiently. Spermatogonial cells grew well when using mitomycin C-treated testicular somatic cells as feeder cells and showed typical characteristic of SSCs. After 13 days of culture,spermatogonial cells formed cell clusters. Immunofluorescence assay showed that SSCs markers glial cell line-derived neurotrophic factor(GDNF)family receptor α1(GFRα1)and VASA protein were highly expressed in the cell clusters. CSCs marker CD44 was expressed in the As,Apr,Aal and the inner cells of the cell clusters,while seldom expressed in the somatic cells.

CONCLUSIONSAn isolation and culture system of SSCs derived from DBA/2 mice was established. CD44 is highly expressed in the early stage of spermatogonial cell development.

Animals ; Biomarkers ; metabolism ; Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Glial Cell Line-Derived Neurotrophic Factor ; metabolism ; Hyaluronan Receptors ; metabolism ; Male ; Mice ; Mice, Inbred DBA ; Spermatogonia ; cytology ; Stem Cells ; cytology

This study established an HPLC fingerprint of Tibetan medicine Shaji Gao from different habitats and lay a foundation for Shaji Gao varieties identification and preparation process. The chromatographic condition was as follow: Agilent zorbax SB-C18 (4.6 mm x 250 mm, 5 μm) eluted with the mobile phases of acetonitrile and 0.4% phosphoric acid water in gradient mode. The flow rate was 1.0 mL x min(-1), and the detection wavelength was set at 360 nm. The fingerprints of 15 batches Shaji Gao were carried out by similarity comparation, 7 chromatographic peaks were extracted as the common peaks of fingerprint, 3 peaks were identified, which were quercetin, kaempferol and isorhamnetin. The similarity degrees of 14 batches of samples were above 0.9 and 1 batch of samples was below 0.9. This is the first established fingerprint of Shaji Gao by using HPLC. This method has good precision, stability and repeatability that it could provide basis for quality control and evaluation of Shaji Gao.
Chromatography, High Pressure Liquid ; methods ; Medicine, Tibetan Traditional ; Quality Control

Objective To evaluate the efficacy of radiosynovectomy with 32P colloid in haemophilic synovitis of adolescents. MethodsRadiosynovectomy with 32P colloid was primary performed on 26 male haemophilic patients(26 joints),whose average age was 16 years(11 to 21 years).The average dose of 32P colloid was 2.1 mCi(1.0 to 3.0 mCi). Results After 6-month interval,haemarthrosis was reduced by no less than 30% in 23 patients,with a total efficacy of 88.5%.The mean frequency of haemarthrosis was reduced from 1.9 per month of presynovectomy to 0.3 per month of postsynovectomy(P

Background The construction of tissue-engineered corneal endothelium needs the functional seeding cells,so how to culture a large amount of functional corneal endothelial cells (CECs) is an urgent problem to be solved.Objective The aim of this study was to evaluate the role of aqueous humor on bovine CECs in vitro.Methods Aqueous humor of 1.2 ml was collected from the anterior chamber of bovine and sterilized,and the liquid supernatant was obtained.The bovine CECs were isolated from bovine cornea and then cultured in low glucose Dulbecco Modified Eagle Medium with 10％ fetal bovine serum (FBS) in vitro.Aqueous humor was added into the medium with the final concentration of 2.5％,5.0％,l0.0％,15.0％ and 20.0％,respectively,and no aqueous humor was added in the control group.Cell counting kit-8 (CCK-8) assay was used to detect the absorbency value of CECs for the evaluation of cell proliferation.Progression of the cell cycle was analyzed by flow cytometry (FCM).After confluence of the cells was reached,1 ml plastic spear tip was used to scratch the cell single layer,and the cells were incubated consequently in medium with 10％ FBS and with or without aqueous humor for 24 hours.Healing area of the cell single layer was measured.The cells were incubated at a density of 6 × 105 cells/ml and cultured using medium with or without 10.0％ aqueous human for 5 days,and the number of the cells was analyzed by DAPI fluorescence technique.Results Under the phase-contrast microscopy,the confluent CECs showed a slabstone-like and hexagonal appearance.CCK-8 assay revealed that the absorbance values of CECs was significantly different among the various culture groups (F=4.051,P =0.007),and the absorbance value in different concentrations of aqueous human culture groups was significantly higher than that in the control group (P ＜ 0.01).FCM showed that the percentage of the cells in S-G2 phases was (34.80-±3.13)％ in the 10.0％ aqueous humors group and (23.06±1.13)％ in the control group,showing a significant difference (t =-5.729,P=0.005).The scratch test showed that the healing area of the cell signal layer was (0.116±0.019) mm2 in the 10.0％ aqueous humors group and (0.358 ±0.049) mm2 in the control group,showing a significant difference (t =13.842,P =0.000).The density of cells in the 10.0％ aqueous humor group was (1439± 1 10)/field,which was more than (1162±45)/field in the control group (t =-11.020,P=0.000).Conclusions Aqueous humor at the concentration of 10.0％ promote the growth and proliferation of bovine CECs.The result suggests that 10.0％ aqueous humor can be used as a promoting agent during the culture of CECs.

This paper described the results of the optimal method for preparation and extraction of lectin from Phaeoporus obliquus(Pers ex Fr).(J.Schroet).We determined the optimal buffer solution using the ag-glutination test with 2% hare blood cells.The optimal extraction process included the following parameters that were conformed by optimization analysis: material-water ratio,extraction time,concentration of sodium chloride and the pH value.The results are that the agglutination indexes of POL using TBS and PBS extrac-tion buffer were 64 and 16,respectively.The optimal extraction process is that,the material-water ratio was 1:50,the extraction time was 20 h,while the concentration of sodium chloride was 0 mol/L and pH was 8.0.The agglutinability of POL was 256 examined by this method.The optimized extraction process is stable and practicable,which may supply some basis for the application of the lectin from Phaeoporus obliquus in immunoregulation.

Objective To investigate the effect of brain-derived neurotrophic factor (BDNF) gene modified neural stem cells (NSCs)transplantation on cerebral ischemic injury in rats.Methods NSCs from newborn rat hippocampus were isolated,cultured in a medium containing fibroblast factor (FGF) in vitro. Their proliferation and differentiation were detected by immunohistochemistry.Virus vectors pLXSN-BDNF were built and BDNF were transfected into NSCs.Biological activity were detected to obtained engineering stem cells of BDNF protein with secretary activity.Middle cerebral artery occlusion (MCAO) models were made and transplanted with NSCs-BDNF by stereotaxic technique.The effect of transplantation on MCAO models was evaluated histologically and behaviorally.Results Statistical analysis showed that the behavioral performance of the animals improved after transplantation (Longa mark being 1.343?0.293 four weeks after stem cell transplantation).The number of hippocampal dentate gyrus neurons increased to 87.5%?6.6% , four weeks after stem cell transplantation on Nissle stained hippocampal sections,which was significantly different from that of controls.Positively BrdU stained neural stem cells revealed by immunohistochemistry in the cultured cells and in the transplanted brain sections,indicated that the engineering cells transplanted had survived in the host brain and began to proliferate.Conclusion Transplantation of BDNF genetically modified NSCs can effectively promote the recovery from cerebral ischemic injury.

Objective To study the clinical significance of expression of survivin gene in transitional mucosa(TM) adjacent to rectal carcinoma.Methods Mucinhistochemical methods were used to observe the distribution of TM adjacent to rectal carcinoma, and using immunohistochemical methods to observe the expression of survivin gene product in TM,normal mucosa,atypical dysplasia and cancer tissue.Results The positive expression level of survivin gene product was lowest in normal rectal mucosa,and gradually increase in TM,dysplasia and carcinoma tissue(all P

Objective To study the effect of the extract of Chrysanthemum Indicum(CI)on the expression of tumor necro-sis factor-α (TNF-α) and neutrophil function in chronic bronchitis (CB)rats and to explore the therapeutic mechanism for chronic bronchitis. Methods The extract of CI was prepared. Wistar rats were randomly divided into blank control group, model group, Chuanxiling group, and low-, middle-and high-dose CI extract groups. Rat model of CB was established by intratracheal injection with low-dose lipopolysaccharide. After drug intervention, the expression of TNF-α in rat serum and bronchoalveolar lavage fluid was detected by ELISA method. Phagocytic function of the neu-trophil and respiratory burst of the rats were measured by flow cytometry (FCM). Results Compared with the blank con-trol group, the phagocytic function of the neutrophil, respiratory burst of the rats, and the expression of TNF-α in rat serum and bronchoalveolar lavage fluid of the model group were increased significantly. CI extract significantly decreased the abnormal rising of the above indexes. Conclusion Down-Regulation of TNF-α expression, and decrease of the neutrophil phagocytic function and rat respiratory burst may be one of the therapeutic mechanisms of Chrysanthemum In-dicum extract for the treatment of CB.

Objective To observe the effects of casein and excessive iodine on histomorphalogY and ultrastructure of mouse thymicL Methods Based on 2 × 3 factorial design,the experimental mice were divided into 6 groupg Animal models were estabhshed by feeding the mice with different levels of iodine water and casein food.The levels of iodine were 50,600 μg/L in drinking water and 0(Ⅰ),10%(Ⅱ),20%(Ⅲ)of casein in food respectively.After 12 months,the thyroid weight was measured and the morphology of thyroid was observed under optical and electron microscope.Results Factorial analysis showed that iodine factors obviously affected the thyroid absolute and relative weiights of mice(F=16.23,9.47,P＜0.01),and there was interaction between casein and iodine(F=5.29,4.68,P＜0.01 or＜0.05).Compared wiht 150Ⅰ[(5.91±0.82)rag,(117.0±22.2)mg/kg]and 50Ⅲ[(4.90±0.63)rag,(106.1±13.3)mg/kg]groups.thyroid absolute and relative weights of the mice increased in 600 Ⅰ[(7.60±2.40)mg,(143.3±43.2)mg/kg]and 600Ⅲ[(8.63±1.88)mg,(166.2±39.4)mg/kg]groups(P＜0.05 or＜0.01),respectively.But compared with 600 Ⅰ and 600Ⅲ groups.they were reduced obviously in 600Ⅱ[(5.76±1.13)mg,(109.8±16.5)mg/kg]group(P＜0.05 or＜O.01).Colloid goiter,lymphocyte infiltration were found,some of the follicles epithelial cells appeared active under light and electron microscope in iodine excels group,which,however,decreased obviously along with the increase of casein dose.Conclusions Long-term excessive iodine may cause colloid goiter and inflammation injury of mice,possibly leading the development of thyroiditis in mice,which may be partly reduced by casein.