1.Optimized effect of connexin 43 gene silencing on the proliferation of fetal liver stem cells
Zengguang ZHANG ; Mingfang QIN
Chinese Journal of Tissue Engineering Research 2015;(14):2133-2137
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.
2.On the teachers'leading role in physiology teaching
Chinese Journal of Medical Education Research 2006;0(11):-
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..
3.TheEffectofIntermitentCyclicEtidronateDisodiumofDifferentDosesonExperimentalOsteoporosisinOvariectomizedRats
Mingfang ZHANG ; Naixin ZHANG ; Yubin TAN ; ETAL
Chinese Journal of Orthopaedics 1996;0(09):-
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.
4.Determination of Fluoroacetamide by Polarography
Mingfang ZHU ; Ning LONG ; Zhengqi ZHANG
Journal of Environment and Health 1993;0(01):-
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.
5.The expression and clinical significance of apoptosis stimulating protein of p53 in nonsmall-cell lung cancer
Shijun LI ; Guangxia SHI ; Mingfang ZHANG ; Mingxia ZHANG
Chinese Journal of Laboratory Medicine 2009;32(11):1246-1250
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.
6.Effects of N-methyl-D-aspartate on the cochlear potentials
Lan ZHANG ; Jinhui XIA ; Mingfang DIAO ; Wenyuan GAO
Academic Journal of Second Military Medical University 2001;22(12):1126-1129
Objective: To study the effects of TNFα on wound healing. Methods: The skin lesion and incisions were made on mice and rhTNFα was applied in both skin lesion and incision. Tissue strips were evaluated for tensiometry and histologic features after healing. Results: It was found that rhTNFα 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 TNFα 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. rhTNFα of suitable doses can promote wound healing. This shows that TNFα, as a wound healing promoter, has the potentials for clinical application.
7.Inducible nitric oxide synthase and nitric oxide in smooth muscle cells of human abdominal aortic aneurysm
Mingfang LIAO ; Zaiping JING ; Xin ZHAO ; Suzhen ZHANG ;
Chinese Journal of General Surgery 2001;0(09):-
Objective To investigate expression of inducible nitric oxide synthase (iNOS) and production of nitric oxide(NO) in cultured vascular smooth muscle cells(SMCs)from human abdominal aortic aneurysm(AAA). Methods TypeⅠ collagenase enzyme digestion method was used to culture AAA SMC . Cell proliferation curves for AAA SMC proliferation ability were evaluated,? smooth muscle actin (? SMA) , iNOS protein expression of AAA SMC and nitrite and nitrate (NO 2 -+NO 3 -,NO X ) in cultured media were measured. Results AAA SMCs had a limited proliferation ability, with 45%?5 8% positive rate of ? SMA and 86 7%?4 6% of iNOS protein expression. High concentration of NO X existed in the cultured media. Conclusion AAA SMCs present abnormal but limited proliferation and changed phenotype; The expression of iNOS and high production of NO of AAA SMCs may play an important role in AAA pathogenesis.
8.Phonological Characteristics and Rehabilitation of Abnormal Velar Consonants in Operated Patients with Cleft Palate
Mingfang HU ; Feng LI ; Lina XU ; Yanyun ZHANG
Journal of Audiology and Speech Pathology 2017;25(4):369-374
Objective To study the phonological characteristics and rehabilitation methods of abnormal velar consonants in operated patients with cleft palate.Methods A total of 110 patients (60 males and 50 females, aged 4 to 27 years with the mean age as 9.01±4.12) with abnormal velar consonants after cleft palate treatment over one month were involved in this study.According to the severity of articulation disorders, the patients were divided into mild, moderate and severe groups who received 1 to 5 weeks of targeted rehabilitation training.The phonological characteristics of abnormal velar consonants and the correction between velar consonant misarticulation and age/gender were analyzed.Results The error rate of /g/ was 96.36%(106/110) which was the highest, then /k/ and /h/ was 74.55%(82/110) and 19.09%(21/110), respectively.Both /g/(80.19%, 85/106) and /h/(76.19%, 16/21) mainly showed omission while /k/ was mainly substituted by /h/(54.88%, 45/82) and /t/(29.27%, 24/82).95 cases were cured and 12 cases improved after specific rehabilitation.The overall cure rate was 86.36%.The cure rates for the mild, moderate and severe groups were 96.00%(24/25), 90.91%(30/33), and 78.85%(41/52),respectively.Males accounted for 51.76%(44/85) and females for 48.24%(41/85) among omission cases;males accounted for 51.47%(35/68) and females for 48.53%(33/68) among substitution cases.There was no significant correction among the number of incorrect words and age(r=-0.140, P>0.05) as well as gender(r=0.090, P>0.1).Conclusion Abnormal velar consonants in operated patients with cleft palate mainly had problems with /g/ and /k/./g/ showed mainly omission errors and /k/ mainly substitution.The targeted rehabilitation methods established in the study are significantly beneficial.
9.Correlation between urinary fluoride level and prevalence of dental caries-the effects of defluoridation for 10 years
Jie XIANG ; Qiushi WANG ; Li YAN ; Mingfang ZHANG ; Quanyong XIANG
Chinese Journal of Endemiology 2015;34(12):884-887
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.
10.The expression of human tissue inhibitor of metalloproteinase-2(TIMP-2)gene in rat aortic smooth muscle cells mediated by adenoviral vector
Xin ZHAO ; Peirui ZHANG ; Xianghai YU ; Mingfang LIAO
Chinese Journal of Current Advances in General Surgery 2004;0(05):-
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.