1.Association of some SNP in TGFA,IRF6 and nonsyndromic cleft lip with or without cleft palate by DNA microarray technology
Ya SHEN ; Lu CHENG ; Wei-Dong WAN ; Zu-Hong LU ; Lian-Ming LIAO ; Yu-Gui CUI ; Jia-Yin LIU ;
Chinese Journal of Laboratory Medicine 2003;0(12):-
Objective To explore the relationship between some single nucleotide polymorphisms (SNP)loci of interferon regulatory factor 6(IRF6)gene,transforming growth faetor-?(TGFA)gene and nonsyndromic cleft lip with or without cleft palate(NSCL/P)in nuclear families consisting of fathers, mothers and affected offspring with NSCL/P from southeast China.Methods Some SNloci of IRF6 and TGFA were detected by applying microarray technology in nuclear families,and then haplotype relative risk (HRR)and transmission disequilibrium test(TDT)were performed.Results There were no significant difference in genotypes and alleles distribution between patients and their parents.The SNP locus——V274I of IRF6 was associated with NSCL/P(HRR:?~2=4.5816,P
2.Differentiation of Human Amniotic Mesenchymal Cells into Cardiomyocyte-like Cells in Vitro
Lu ZHANG ; Ning FANG ; Dai-Xiong CHEN ; Zu-Lin LIU ; Jin-Wei LIU ; Wei-Hong WAN ; Tao ZHANG ;
China Biotechnology 2006;0(12):-
To explore the plasticity of human amniotic mesenchymal cells(hAMCs) into cardiomyocyte-like cells,hAMCs were isolated from human amnion with collagenase digestion.Phenotype of the isolated cells was analyzed by flow cytometry(FCM).hAMCs were treated with 5-azacytidine and basic fibroblast growth factor(bFGF) to investigate their ability of differentiation into cardiomyocytes.The induced differentiated cells were evaluated by immunofluorescence for desmin and ?-actin expression and by RT-PCR for Nkx2.5,GATA-4 and alpha-myosin heavy chain(?-MHC) mRNA expression.The results showed that,after primary culture,hAMCs could reach a confluence of 80% with swirl like growth at 6 days.The cells proliferated rapidly after passages with a 100% confluence at 3~4 d.hAMCs were positive expression of CD44 and vimentin,but negative for CK19.After induced differentiation at 8~10d,the differentiated cells have close-up arranged with long spindle-shape.At 2 weeks and 4 weeks,induced cells expressed ?-actinin and cardiac-specific transcription factor Nkx2.5.Expressions of GATA-4 and desmin can be detected but ?-MHC can not in the hAMCs both before and after the induction.In conclusion,hAMCs have the ability of differentiation into cardiomyocyte-like cells,which means that hAMCs can be regarded as candidate cells for cellular cardiomyoplasty(CCM).
3.Clinical and electrophysiological evaluation of nocturnal splinting on carpal tunnel syndrome
Chao WENG ; Hong CHU ; Bin PENG ; Ying-Bing KE ; Wen-Hua WANG ; Hong-Juan DONG ; Zu-Neng LU
Chinese Journal of Neuromedicine 2010;09(11):1132-1136
Objective To assess the effectiveness of nocturnal splinting on carpal tunnel syndrome (CTS) by clinical scores and nerve conduction studies (NCS), and explore their correlations.Methods Forty-one patients (64 wrists), chosen from 66 consecutive patients with CTS from April 2009 to January 2010 meeting the inclusion criteria, were enrolled. The enrolled subjects were clinically evaluated by symptom severity scale (SSS) and functional status scale (FSS), and electrophysiologically evaluated by conventional nerve conduction studies (NCS); distal motor latency (DML) of wrist-abductor pollicis brevis, sensory conduction velocity (SCV) of wrist-index finger and wrist-ring finger, and the differences of distal sensory latency between the median and ulnar nerves (△DSL) were measured. The patients were instructed to use each splint on dorsal and palmar surface of the hand, centered at the distal wrist crease, to fix the wrist in neutral posture at bedtime. SSS, FSS and NCS were evaluated before splinting and (3.03±1.16) months after splinting; the follow-up was completed in 29 patients (31 wrists).Results (1) The abnormality rates of DML, wrist-index finger SCV, wrist-ring finger SCV and△DSL were 85.9%, 78.1%, 81.3% and 96.9%, respectively. (2) The SSS scores (1.55±0.38), FSS scores (1.40±0.27) and△DSL (1.24±0.61) after splinting was significantly decreased as compared with those before splinting (1.77±0.38, 1.53±0.31, 0.97±0.60); and the DML [4.14±0.76 (ms)] after splinting was significantly shortened as compared with that before splinting [4.53±1.25 (ms)]. No improvement of clinical scores was noted in 9 patients (14 wrists, 45.8%) after splinting. (3) The SSS scores were less significantly correlated to DML (r=0.420, P=0.019), wrist-index finger SCV (r=-0.425, P=0.017),wrist-ring finger SCV (r=-0.519, P=0.003), and no correlation between SSS scores and△DSL was noted (r=0.189, P=0.309); no correlation between FSS scores and the parameters of NCS was found either (P>0.05). Conclusions Splinting is effective at least in a short-term in more than halfpatients with CTS.Little correlation is noted between clinical scores and NCS, suggesting that utilization both approaches to assess the therapeutic effect is of more significance. △DSL is the most sensitive parameter in the electrodiagnosis of CTS.
4.Using protein chips to study mechanism underlying reversion of drug resistance in leukemia cells in tetrandrine alone or in combination with droloxifene.
Bao-An CHEN ; Juan DU ; Chun-Xiu ZHANG ; Jian CHENG ; Feng GAO ; Zu-Hong LU
Journal of Experimental Hematology 2005;13(6):999-1003
The aim of this study was to investigate the mechanism to reverse the drug resistance of leukemia cells in tetrandrine (Tet) alone or in combination with droloxifen (Drol) by using protein chips and to lay the theoretical basis for the clinical applications. Three monoclonal antibodies against P-glycoprotein (P-gp), the multidrug resistance-associated protein (MRP1) and the breast cancer resistance protein (BCRP) were immobilized onto the agarose gel film-coated glass slides. Protein chips were prepared respectively from K562/A02 cells cultured for 12, 24 and 48 hours with Tet alone or in combination with Drol. The results showed that Tet alone or in combination with Drol could decrease only the expression of P-gp in a time-dependent manner, the effect for 48 hours as follows: Tet + Drol 82.620 +/- 3.227; Tet alone 86.440 +/- 2.906; Drol alone 87.230 +/- 2.049; control 93.670 +/- 2.748 (P < 0.05). However, down-regulation of P-gp by K562/A02 cells cultured with Tet alone or in combination with Drol began at 24 hours (Tet + Drol 85.270 +/- 3.095; control 93.670 +/- 2.748, P < 0.05). The results were coincident with that of FCM. It is concluded that Tet and Drol can downregulate the expression of P-gp in the time-dependent way. There is a significant difference between Tet alone and Tet combined with Drol at 24 hours (P < 0.05). The expression of MRP1 and BCRP are not closely correlated with the reversal mechanism of Tet and Drol, and which may be involved in the mechanism of this combination to reverse multidrug resistance in leukemia.
ATP Binding Cassette Transporter, Sub-Family G, Member 2
;
ATP-Binding Cassette Transporters
;
biosynthesis
;
ATP-Binding Cassette, Sub-Family B, Member 1
;
biosynthesis
;
Antineoplastic Agents
;
pharmacology
;
Benzylisoquinolines
;
pharmacology
;
Drug Resistance, Multiple
;
Drug Resistance, Neoplasm
;
drug effects
;
Drug Synergism
;
Humans
;
K562 Cells
;
Leukemia
;
metabolism
;
pathology
;
Multidrug Resistance-Associated Proteins
;
biosynthesis
;
Neoplasm Proteins
;
biosynthesis
;
Protein Array Analysis
;
Tamoxifen
;
analogs & derivatives
;
pharmacology
5.Percutaneous coronary intervention combined cardiac resynchronization therapy for refractory heart failure secondary to ischemic cardiomyopathy.
Ya-ling HAN ; Hong-yun ZANG ; Dong-mei WANG ; Quan-min JING ; Shou-li WANG ; Zu-lu WANG
Chinese Journal of Cardiology 2005;33(1):17-21
OBJECTIVETo evaluate the efficacy and safety of percutaneous coronary intervention (PCI) combined cardiac resynchronization therapy (CRT) for refractory heart failure secondary to ischemic cardiomyopathy (ICM).
METHODSPCI and CRT were performed in 7 ICM patients confirmed by angiography with NYHA class IV, QRS duration >/= 130 ms in 6 of them, III degrees AVB in 1 patient, fast ventricular heart rate Af in 1 patient, ventricular fibrillation history in 2 patient. All of them had their LVEDD >/= 55 mm, and LVEF = 0.40 detected by UCG. PCI was performed first in 5 patients, and their follow-up angiography showed no restenosis 6 months after PCI, then CRT was given. CRT was performed first in 2 patients and 2 weeks later PCI was combined.
RESULTSThe procedures of PCI and CRT were performed successfully in all patients. Five patients received right atrial and biventricular pacing, one patient with Af received biventricular pacing and atrial-ventricular node radiofrequency ablation at the same procedure, and the another one patient received CRTD. One out of seven patients died of re-AMI 4 months after the combination therapy, and the other 6 patients had been alive 5 - 41 (23.2 +/- 13.8) months during the follow-up period. The heart function of the 7 patients had further improved after PCI and CRT combined therapy compared to that of PCI or CRT only. Their NYHA class decreased from IV to II, 6-minute walking distance increased steadily, and mitral regurgitation reduced and QRS duration shortened significantly. The LVEDD decreased and LVEF increased significantly in 2 patients without ventricular aneurysm, and slight improvement or no change were in the other 5 patients.
CONCLUSIONFor patients with refractory heart failure secondary to ICM, the combination of PCI and CRT could obviously improve their heart function, quality of life and prognosis, which also very safe in perforation.
Adult ; Aged ; Angioplasty, Balloon, Coronary ; Cardiac Pacing, Artificial ; methods ; Combined Modality Therapy ; Female ; Heart Failure ; etiology ; therapy ; Humans ; Male ; Middle Aged ; Myocardial Ischemia ; complications ; therapy ; Treatment Outcome
6.The induction apoptosis of HL-60 cells by low molecular weight compounds of taurine, ornithine and carnosine from new born calf liver.
Jin-hong ZHANG ; Qian LU ; Wen-jing SHI ; Zu-ze WU ; Li-sheng WANG
Chinese Journal of Applied Physiology 2005;21(2):200-205
AIMClinical studies stated that low molecular weight compounds (< 1.0 kd) extracted from the new born calf liver could effectively inhibit the proliferation of tumor cells. In this report, we observed inhibition effects and their regulative mechanisms of taurine, ornithine, carnosine on the proliferation of HL-60 cells.
METHODSThree active ingredients, i.e., taurine, ornithine and carnosine were separated by ion-exchange chromatographic column and identified from the low molecular weight filtrate of new born calf liver. MTT assay was used to test the survival rate of HL-60 cells and normal lymphocytes treated by the three ingredients. The various effects of the three compounds on HL-60 cells were respectively evaluated by agarose gel electrophoresis, ESR and immunohistochemical methods.
RESULTSThese compounds effectively inhibited the proliferation of HL-60 cells and induced apoptosis which was determined by apoptotic changes in morphology and nuclear DNA degradation. Whereas no inhibition effects on normal lymphocytes were observed. In addition, the results of ESR showed that the activity of oxygen radical within HL-60 cells treated with there compounds decreased to trace level. Furthermore, in the immunohistochemical experiments, we found that the level of p45/skp2 in HL-60 cells decreased while the level of p27/kip increased.
CONCLUSIONThe taurine, ornithine and carnosine compounds can selectively suppress tumor cells proliferation by regulating the level of cell cycle proteins.
Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Carnosine ; pharmacology ; Cattle ; HL-60 Cells ; Humans ; Liver ; chemistry ; Ornithine ; pharmacology ; Taurine ; pharmacology
7.XPC Lys939Gln polymorphism is associated with the decreased response to platinum based chemotherapy in advanced non-small-cell lung cancer.
Xiao-Li ZHU ; Xin-Chen SUN ; Bao-An CHEN ; Ning SUN ; Hong-Yan CHENG ; Fan LI ; Hong-Ming ZHANG ; Ji-Feng FENG ; Shu-Kui QIN ; Lu CHENG ; Zu-Hong LU
Chinese Medical Journal 2010;123(23):3427-3432
BACKGROUNDPlatinum-based chemotherapeutics are the most common regimens for advanced non-small-cell lung cancer (NSCLC) patients, and genetic factors are thought to represent important determinants of drug efficacy. We prospectively assessed the status of the XPC Ala499Val and Lys939Gln gene polymorphisms and investigated whether these SNPs can predict the response to cisplatin/carboplatin-based regimens in advanced NSCLC patients in a Chinese population.
METHODSThe treatment outcomes of 96 advanced NSCLC patients who were treated with platinum-based chemotherapy were evaluated. The polymorphic status of xeroderma pigmentosum group C (XPC) gene was genotyped by the 3-D polyacrylamide gel-based DNA microarray method.
RESULTSThe distributions of XPC Lys939Gln genotypes differed significantly between the response group (complete + partial responses) and the non-response group (stable + progressive disease; P = 0.022). The heterozygous A/C genotype carriers had a poorer response rate than the wild A/A genotype carriers in stage III (OR, 0.074; 95%CI, 0.008 - 0.704; P = 0.023). The XPC Ala499Val polymorphisms were not associated with response to platinum-based chemotherapy.
CONCLUSIONPolymorphisms of the XPC gene, Lys939Gln, may be a predictive marker of treatment response for advanced NSCLC patients in stage III.
Adult ; Aged ; Alleles ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carboplatin ; administration & dosage ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; genetics ; pathology ; Cisplatin ; administration & dosage ; DNA-Binding Proteins ; genetics ; Female ; Genotype ; Humans ; Lung Neoplasms ; drug therapy ; genetics ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Polymorphism, Single Nucleotide ; Prospective Studies
8.Analysis of 322 Cases of 2-Year-Old Children Testing Results with Bayley Scales of Infant Development in Nanjing
xia, CHI ; qin, HONG ; tian-hong, YAO ; ya-qin, XU ; jia-zhen, DAI ; min, ZHANG ; ning-chuan, CHEN ; xi-rong, GUO ; mei-ling, TONG ; xiao-yan, KE ; hui-hua, DENG ; zu-hong, LU
Journal of Applied Clinical Pediatrics 2004;0(11):-
Objective To provide scientific evidences for improving children early integrated development,through analyzing the mental developmental status and characteristics of 322 cases of 2-year-old children in Nanjing.Methods Intelligence and motor development condition in 322 cases of 2-year-old children were assessed by using Bayley Scales of Infant Development test,and the assessed results were analyzed.Results 1.The incidences of the children whose mental development index(MDI)or psychomotor development index(PDI)were under 69 were 3.1% and 5.6%,respectively;2.The MDI mean score(114.34?19.65)was significantly higher than that of PDI(101.73?21.53)(t=9.71,P0.05).Conclusions The incidences of mental retardation in this study were consistent with the result reported by World Health Organization.There were differences between motor and intelligence development in children,as well as the intelligence development between male and female.Therefore,it should be implemented early childhood developmental screening in child health care.Parents should be given scientific guides about intelligence and motor development of children.
9.Study on the genotyping of single nucleotide polymorphisms for a large number of samples by three-dimensional polyacrylamide gel-based microarray method.
Lu CHENG ; Peng-feng XIAO ; Bei-li SUN ; Qin-yu GE ; Zu-hong LU
Chinese Journal of Medical Genetics 2009;26(3):293-297
OBJECTIVETo genotype single nucleotide polymorphisms (SNPs) in a large number of samples by applying three-dimensional polyacrylamide gel-based microarray.
METHODSThe method relies on copolymerization of acrylamide-modified PCR products with acrylamide monomers and acryl-modified slides to prepare gel-based microarray. Then array is hybridized with a pair of specific probes and the two universal dual-color fluorescent detectors labeled with Cy3 or Cy5 respectively (Tag1 and Tag2). Electrophoresis is used in post-hybridization to remove the nonspecifically bound targets and mismatches. Finally, genotyping is based on the images captured through two-color fluorescent scanning.
RESULTSThe 3-D gel-immobilization of nucleic acids has a high immobilization yield and good hybridization efficiency. As universal dual-color fluorescent detectors are used, it is not required that specific probes be labeled for all SNPs, therefore the expense for synthesis can be reduced considerably. Electrophoresis in post-hybridization can enhance the capability for discriminating a single nucleotide mismatch from the perfectly matched sequence and improve the signal-to-noise ratio significantly.
CONCLUSIONThe gel-based microarray is a rapid, simple and high-throughput method for SNPs genotyping and may be very competitive in the efficiency, fidelity and cost for constructing DNA microarrays, which will hold significant promise for applications in human DNA diagnostics.
DNA Mutational Analysis ; methods ; DNA Probes ; Electrophoresis, Polyacrylamide Gel ; methods ; Fluorescent Dyes ; Genotype ; Humans ; Molecular Diagnostic Techniques ; Nucleic Acid Hybridization ; methods ; Oligonucleotide Array Sequence Analysis ; methods ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide
10.Influence of hepatocyte growth factor on biological characteristics of bone marrow-derived mesenchymal stem cells.
Hong-Jun LIU ; Hai-Feng DUAN ; Zhuo-Zhuang LU ; Hua WANG ; Qun-Wei ZHANG ; Zu-Ze WU ; Li-Sheng WANG
Journal of Experimental Hematology 2005;13(6):1044-1048
Hepatocyte growth factor (HGF) is one of major growth factors in the bone marrow microenvironments with which the proliferation, differentiation and migration of bone marrow-derived mesenchymal stem cells were closely contacted. However, its roles in the regulation of proliferation, differentiation and migration of bone marrow-derived mesenchymal stem cells remain unclear. This study was aimed to investigate the effect of HGF on biological characteristics of bone marrow-derived mesenchymal stem cells. Expression of c-Met, the receptor for HGF was detected by immunohistochemistry assay, cell proliferation was determined by MTT, activity of ALP was quantitatively assayed, cell migration and anoikis-induced MSC apoptosis were analyzed. The results showed that HGF not influenced the proliferation and osteogenic differentiation of bone marrow-derived mesenchymal stem cells. Treatment of bone marrow-derived mesenchymal stem cells with recombinant human hepatocyte growth factor resulted in inhibition of anoikis-induced apoptosis. HGF significantly stimulated the migration of bone marrow-derived mesenchymal stem cells. Both PI-3 kinase and MAPK kinase were proved to be involved in HGF-induced migration. It is concluded that HGF/c-Met signal regulates the apoptosis and migration of bone marrow-derived mesenchymal stem cells.
Anoikis
;
drug effects
;
Bone Marrow Cells
;
cytology
;
drug effects
;
metabolism
;
Cell Movement
;
drug effects
;
Cell Proliferation
;
drug effects
;
Cells, Cultured
;
Hepatocyte Growth Factor
;
pharmacology
;
Humans
;
Immunohistochemistry
;
Mesenchymal Stromal Cells
;
cytology
;
drug effects
;
metabolism
;
Proto-Oncogene Proteins c-met
;
biosynthesis