BACKGROUND:Fetal liver stem cel s have the potential to differentiate into hepatocytes and bile duct cel s, and participate in the repair and reconstruction of the liver, which is an important source of hepatocytes. But there are a little amount of fetal liver stem cel s in human body, and how to obtain a certain number of high-purity fetal liver stem cel s is currently a hot research. OBJECTIVE:To construct a siRNA carrier that can effectively inhibit the expression of connexin 43 (Cx43) in rat fetal liver stem cel s, and to investigate the effect of Cx43 inhibition on the proliferation and cel cycle of fetal liver stem cel s cultured in vitro. METHODS:Fetal liver stem cel s were cultured by the suspension culture in vitro, siRNA sequences targeting Cx43 (Cx43-siRNA) and negative control sequence (NC-siRNA) were designed and synthesized. Then, rat fetal liver stem cel s were transferred electrophoretical y and divided into three groups:blank group, NC-siRNA group, Cx43-siRNA group. Real-time PCR and western blot were used to assess the knockdown efficiency. Cel ular proliferation was determined by cel growth curve and cel counting kit-8 assay. The cel cycle was analyzed by flow cytometry. RESULTS AND CONCLUSION:After transfection, the Cx43 gene and protein expression levels were declined dramatical y in the Cx43-siRNA, NC-siRNA and blank groups, and the cel s grew faster. The number of cel s at G0/G1 phase decrease, but the number of cel s in S phase increased. There were significant differences between the groups (P<0.05). Electrophoretic transfer of Cx43-siRNA can promote the proliferation of cultured fetal liver stem cel s and optimize the cel culture.

The combination of teachers'leading role and the students'main role is a basic principle of teaching. This paper discusses how to play the leading role of teachers from pre-teaching preparation and classroom teaching to coaching students and answering the questions after school..

Thetherapeuticeffectsofintermitentcyclicetidronatedisodium(HEBP)ofdifferentdosesonovariectomy-inducedosteoporosiswerestudied.53femaleWistarrats,weighingwithanaver-ageof266gmwererandomlydividedinto5groups.Gourp1hadbilateralovariectomy(OVX)andwasgivenlowdoseHEBPof25mg/kgBW/day;group2hadOVXandgivenmediumdoseHEBPof36mg/kgBW/day;group3hadOVXandgivenhighdoseHEBPof51mg/kgBW/day;group4hadOVXandgroup5hadshamOVX.HEBPwasadministratedoralyduringfastingfor5daysfolowedbyarestperiodof25daysandtheregimewasrepeatedfor3times.Alanimalsweresacrificedat90thdayafterOVX.RadioimmunoassayofserumestradiolconfirmedthesuccessofOVX.TheserumCaandPcon-tent,AKPaseactivity,femoralrelativedryweight,CaandPcontentwereanalysedrespectively.Therighttibiaewereprocessedundecalcifiedforquantitativebonehistomorphometry.TheresultsshowedthatthebonecalciumcontentandtrabecularbonevolumeofOVX-ratstreatedwithHEBP,especialywiththelowdosegroupwerehigherthanthatofthesimplyOVX-ratandtheboneturnoverparameterssuchasAKPaseactivity,boneformationsurface,boneresorptionsurface,activeformationsurface,bonemineralappositionrateandboneformationratewerelowerthanthoseofthesimplyOVX-rat.TheresultsindicatetheHEBPcouldretardthedegreeofosteopososisthroughreducingtheelevatedboneresorptionandboneturnoverinducedbyOVX.Meanwhile,theresultssuggestthattheefectoflowdoseHEBP(25mg/kgBW/day)wasmoreefectivethanthatofthehighdose.

Objective To establish a polarographic method for detecting fluoroacetamide.Methods On the condition of strong basic solution with high temperature fluoroacetamide was decomposed to form a decomposed product ALC-La-F - complex.In a pH4.90buffer solution containing KNO 3 and hexamethylenetetramine,the decomposed product was determined with com-plex-adsorptive wave at-0.67V by polarography.Re sults The detection limit and linear range of the method was2.0?10 -8 mol/L and5.0?10 -8 -1.2?10 -6 mol/L respectively.The regressive equation was y=3.49x+0.042with a correlation coefficient r=0.9996.The method had been applied to the determination of fluoroacetamide in wheat flour samples satisfyingly.Conclu-sion This method was simple and suitable for the determination of fluoroacetamide in flour.

Objective To investigate the expression and regulation of ASPP family mRNA in NSCLC cell lines.and study the diagnostic and therapeutic significances of apoptosis stimulating of ps3 expression in NSCLC Patients.Methods Real-time fluorescence quantitative PCR wag established and used to measure the expression Ievels of ASPP mRNA in NSCLC cell lines A549(wild-type p53)and NCI-H157 (mutant-type p53)with p53 different genetic background,and ASPP mRNA was also detected in NSCLC tissue,peripheral blood of 37 NSCLC patients.Western Bloting was used to examine the levels of ASPP1 and ASPP2 proteins of NSCLC cell Iines.Chemosensitivity of the cell lines to CDDP was analyzed by MTT assay,and detect the change of ASPP1 and ASPP2.Compared with the ASPPs mRNA expression between NSCLC patients and healthy controls.and monitoring ASPPs mRNA expression in patients with NSCLC chemotherapy.Results The slope rotes of standard curves were-3.249,-3.358 respectively.and correlation coefficients were more than 0.98.Amplification efficiency of standard curves were 1.156.1.028 respectively.The IC_(50) values of NCI-H157 and A549 cells were 3.70.10.48 μg/ml respectively.Mean ASPP1 ASPP2 mRNA levels were a statistically significant 2.66 folds and 6.98 folds higher in NCI-H157 cell compared to A549 cell The levels of ASPP1 mRNA in tumor tissues and peripheral blood were (4.27±0.57)×10~3,(2.49±0.32)×10~3 in patients with NSCLC respectively,and the levels of ASPP2 mRNA in tumor tissues and peripheral blood were(2.34±0.75)×10~3,(7.00±1.17)×10~3 in Patients with NSCLC respectively.The levels of ASPP1 and ASPP2 mRNA in tissues and peripheral blood were(1.32±0.21)×10~4,(1.46±0.31)×10~4 and(1.38±0.19)×10~4,(1.28±0.18)×10~4 in control groups respectively.The ASPP1,ASPP2 mRNA levels in patients with NSCLC were significantly lower than those of control groups(t=2.58,3.94,3.62,3.76,P＜0.05).Before chemotherapy the levels of ASPP1 and ASPP2 mRNA in patients with NSCLc were(2.34±0.56)×10~3 and(6.64±0.72)×10~3,and after 2 periods of postoperative chemotherapy the levels were(3.66±0.64)×10~3 and(9.42±0.44)×10~3,the expression was higher than those of ehemotherapy before(t=3.02,4.50,P＜0.05).Conclusions The expression of ASPP1.ASPP2 mRNA in p53 mutant-type cell lines is higher than that in p53 wild-type cell lines.The expression of ASPP maybe has close relationship with chemotherapeutic sensitivity of NSCLC.In NSCLC patient chemotherapy process,the change of ASPP1 and ASPP2 mRNA expression may be related with chemotherapy medicine function.Enhance the expression of ASPP1 and ASPP2 will be beneficial to the tumot treatment.

Objective: To study the effects of TNFα on wound healing. Methods: The skin lesion and incisions were made on mice and rhTNFα was applied in both skin lesion and incision. Tissue strips were evaluated for tensiometry and histologic features after healing. Results: It was found that rhTNFα markedly improved wound healing both in skin lesion and incision wound model, it also markedly enhanced the breaking strength of treated incision and increased wound cellularity in rh TNF α treated groups. Conclusion: The results suggest that TNFα play an important role in regulating inflammatory reactions of local injured tissues and it may improve the functions of fibroblasts, mononuclear phagocytes and endothelial cells. rhTNFα of suitable doses can promote wound healing. This shows that TNFα, as a wound healing promoter, has the potentials for clinical application.

Objective To investigate phonological characteristics and rehabilitation training approaches of bilabial consonant articula-tion disorders in patients after repaired cleft palate. Methods From January, 2012 to January, 2016, 51 patients with velopharyngeal incom-petence (VPI) and 57 patients with velopharyngeal competence (VPC) after cleft palate repaire were enrolled. Their phonological character-istics of bilabial consonants articulation disorders were analyzed. They accepted specific rehabilitation training. The correlation between bila-bial consonants articulation disorders and age was analyzed. Results/b/ showed mainly weak pressure in both VPI and VPC patients,/p/showed mainly weak pressure in VPI patients,/p/was mainly substituted by/b/in VPC patients. 46 cases were cured and 2 cases improved in VPI patients after 1-6 weeks of rehabilitation, and the overall cure rate was 90.20%(46/51), and was respectively 100%(13/13), 93.74%(15/16), 77.27%(17/22) in those with mild, moderate and severe disorder. 53 cases were cured and 4 cases improved in VPC patients after 1-5 weeks of rehabilitation, the overall cure rate was 92.98%(53/57), and was respectively 100%(17/17), 90.48%(19/21), 89.47%(17/19) in those with mild, moderate and severe disorder. The number of incorrect words negatively correlated with age in VPC patient (r=-1.000, P<0.001). Conclusion Bilabial consonant articulation disorders occurs mainly on/b/and/p/in patients repaired cleft palate,/b/shows main-ly weak pressure,/p/shows mainly weak pressure and substitution. Specific speech rehabilitation training is significantly effective.

Objective:To construct an adenoviral vect or carrying human tissue inhibitor of metalloproteinase-2(TIMP-2)gene in order to mediate the expression of TIMP-2 gene in vascular smooth muscle cells(SMC)i n vitro. Methods:A recombinant adenovirus(AdhTIMP-2)containging human TIM P-2 cDNA fragment was generated by homologous recombination in BJ5183 bacteria. Recombinant plasmids were screened by alteration of antibiotics.The adenovirus v ector was then packaged and amplified in 293 cells.The SMC of rat aortic were is olated and cultured in vitro and been infected with AdhTIMP-2.The expression of TIMP-2 was detected by the techniques of Western blot and RT-PCR. Results:The recombinant adenoviral vector carrying human TI MP-2 was constructed. The titer was 4?1011efu/ml after purification.The AdhTIMP-2 could infect the cultured VSMC efficiently(MOI=100,infection ra te=94%).The expression of TIMP-2 gene in those infected cells was detected by R T-PCR.After the cells were infected with AdhTIMP-2 24hours,TIMP-2 protein cou ld be detected in the conditioned medium by Western blot. Conclusion:The recombinant adenoviral vector carrying human TIMP -2 is successfully constructed and AdhTIMP-2 can efficently mediate the expres sion of TIMP-2 gene in cultured VSMC,paving the way for further application in vascular disease gene therapy.

Objective To discuss the application values of image reconstructive technique in 16-slice helical CT scanning. Methods 32 cervical spinal cord lesion images which have processed in MPR, MIP and VR were reviewed to discover the best image reconstructive form. Results By grading, reconstruction score of MPR, MIP and VR was 4.75, 1.75 and 4.5, MPR and VR in top. Conclusion 16-slice CT scanning with MPR and VR image reconstructive of cervical spinal cord can be satisfied the clinical demands.

Objective To study the relationship between the level of fluoride in urine and the prevalence of dental caries in children before and after the defluoridation, and to provide a basis for assessment of the effects of defluoridation projects and for control of dental caries.Methods Wamiao Village, in Jiangsu Province, a formerly severe endemic fluorosis area, was selected to carry out the study.All children aged 8-13 years old before the defuoridation were investigated from September to November in 2002, and children 8-10 years old who were born after the defluoridation were investigated from September to November in 2013.Urinary fluoride level of the children and the prevalence of dental caries were determined, and their relationships were analyzed.The urina sanguinis samples of children were collected in the morning, and the urine fluoride level was tested using the fluoride ion selective electrode.Dental caries diagnoses was referenced to Dental Caries.Results Totally children's urine samples were 236 and 68 respectively before and after defluoridation.Urinary fluoride level of the children was significantly decreased from (3.53 ± 1.81)mg/L (before defluoridation) to (1.39 ± 0.66)mg/L (after defluoridation, t =9.506, P ＜ 0.01);the prevalence of dental caries was increased from 52.73％ (29/55, before defluoridation) to 63.24％ (43/68, after defluoridation), however, the difference was not significant (x2 =1.383, P ＞ 0.05).The DMFT increased from 1.18 (before defluoridation) to 1.68 (after defluoridation), and the epidemic levels of dental caries were all at lower levels.The relationship between urine fluoride level and the prevalence of dental caries as well as the DMFT before defluoridation was a U-shape dose-response curve;which was gone after defluoridation.Conclusions The urinary fluoride level is significantly decreased after defluoridation for 10 years, the prevalence of dental caries is increased but not significantly.The results of this study indicate that the measure of fluoridation to prevent dental caries needs to be further validated.