0.05).Cmax of domestic and imported rebamipide tablet were (0.56?0.24) and (0.59?0.29)mg?L-1, with that tmax were (1.75?0.92) and (1.98?1.05)h,t1/2(?) were (1.86?1.38) and (1.93?1.45)h,AUC0～∞ were (2.48?1.06) and (2.62?1.35)mg?h?L-1. Conclusion The pharmacokinetics of the two products are similar.

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).

OBJECTIVETo study inhibitory efficacy of combined gene therapy for malignant gliomas transfected with antisense human telomerase reverse transcriptase (hTERT)/PTEN in vitro and in vivo.

METHODSTo construct two adenovirus recons which contained antisense hTERT and wild-type PTEN respectively with high performance homologous recombination system in bacteria. The two adenovirus recons were transfected into U251 human malignant glioma cells combinedly or respectively in vitro and in vivo. U251 cell proliferation in vitro was determined by MTT assay and flow cytometry, tumor growth in vivo was measured by the volume of glioma in nude mice. Telomerase activity was detected by telomeric repeat amplification protocol (TRAP) assay. Expression of hTERT and PTEN protein was detected by Western blotting methods.

RESULTSAfter transfection in vitro, the growth of U251 cells was inhibited significantly. The inhibitory effect was time-dependent. The strongest inhibition was observed in combined transfection group, at the 6th day, the survival rate was 37.6%, telomerase activity (only 28.8TPG) was inhibited significantly, hTERT protein expression was inhibited significantly too, which was 0.2106, but PTEN protein expression was increased significantly, which was 0.9630. In vivo, the growth of tumors was also effectively inhibited.

CONCLUSIONGrowth of malignant glioma cells is effectively inhibited after transfection with combined antisense hTERT and PTEN in vitro and in vivo.

Adenoviridae ; genetics ; Animals ; Apoptosis ; Blotting, Western ; Brain Neoplasms ; pathology ; therapy ; Cell Line, Tumor ; Cell Proliferation ; DNA, Antisense ; genetics ; metabolism ; Flow Cytometry ; Genetic Therapy ; methods ; Glioma ; pathology ; therapy ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Mice ; Mice, Nude ; Microscopy, Fluorescence ; PTEN Phosphohydrolase ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Telomerase ; genetics ; metabolism ; Transfection ; Tumor Burden ; Xenograft Model Antitumor Assays

OBJECTIVETo construct a recombinant lentivirus RNA interference (RNAi) vector carrying hTERT gene, and to obtain the titer of the lentiviral stock for investigating the expression in the eukaryotic cells and the effect on the hTERT gene silencing in the eukaryotic cells.

METHODSTwo complimentary oligos of small interference RNA (siRNA) with hairpin structures targeting the hTERT gene and a negative control were synthesized, then ligated with pLVTHM vector and sequenced. The recombinant vectors were then transfected with viral packaging mix into T293 cells, viral supernatant was harvested to determine the titer. U87 cells infected by virus were harvested and the expression of hTERT, telomerase activity and apoptosis were detected by reverse transcription-PCR(RT-PCR), TRAP assay and flow cytometry separately.

RESULTSSequencing data showed that the constructed plasmids contained the correct sequences of hTERT siRNA transcript templates. A vector producing cell line T293 was established, and the titer for transfection was obtained. RT-PCR and TRAP flow cytometry analyses demonstrated that hTERT shRNA expression construct could suppress the expression of hTERT and telomerase activity and induce apoptosis.

CONCLUSIONA lentivirus RNAi vector targeting hTERT gene was successfully constructed, which decreased the expression of hTERT and telomerase activity effectively and induced apoptosis. It has set up a research platform for the gene therapy of tumors which take hTERT as the target.

Base Sequence ; Cell Line ; Flow Cytometry ; Gene Knockdown Techniques ; methods ; Genetic Vectors ; genetics ; Humans ; Lentivirus ; genetics ; Molecular Sequence Data ; Oligodeoxyribonucleotides ; genetics ; metabolism ; Plasmids ; genetics ; metabolism ; RNA Interference ; RNA, Small Interfering ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Telomerase ; biosynthesis ; genetics

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

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.

OBJECTIVETo explore the synthetical index for diagnosing schistosomiasis with ultrasound and to assess the prevalence rate with the index.

METHODSUltrasound indexes of schistosomiasis Japonicum were analyzed by principal component analysis, and the synthetical indexes were assessed by ROC curve.

RESULTSAmong the abnormal rates of the 6 indexes, the lowest was 1.6% comparing with the highest of 59.5%. Significant difference was noficed among the abnormal rates (chi(2) = 631.1, P < 0.01). The individual correlation of the six indexes to each other as will as with age distribution was significant (P < 0.05). The three principal components reflected the degree of pathological changes on liver and spleen. The first principal component was the factor reflecting the degree of liver pathological changes, and the second and third principal components reflected the degree of pathological changes on spleen. The synthetical index D(1) = 0.047X(1) + 0.428X(2) + 1.247X(3) + 0.095X(4) + 0.002X(5) + 0.213X(6) - 12.837 was found by adding the three weight principal components, and it's area under the ROC curve was 0.957. When -1.70 was taken as the critical value, the abnormal rate of population was 66.3%, close to the resident's actual prevalence rate 66.9%.

CONCLUSIONUltrasonography was considered as a method which could rapidly assessing the resident's prevalence rate in the endemic areas of schisitosomiasis Japonicum, and could also provide powerful information for development of strategy on chemotherapy.

Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Prevalence ; Principal Component Analysis ; ROC Curve ; Schistosomiasis japonica ; diagnostic imaging ; epidemiology ; Ultrasonography

AIM To establish a UPLC-MS/MS method for the simultaneous content determination of ellagic acid,gallic acid,dehydrodiisoeugenol,methyleugenol,agarotetrol,elemicin,chlorogenic acid,geniposide,rutin,ferulic acid,hydroysafflor-yellow A,deoxycholic acid,cholic acid,borneol and taurine in Zhuriheng Dropping Pills.METHODS The analysis of 50%methanol solution of this drug was performed on a 30℃thermostatic Thermo C18 column(2.1 mm×100 mm,1.9 μm),with the mobile phase comprising of acetonitrile-0.1%formic acid(containing 10 mmol/L ammonium acetate)flowing at 0.3 mL/min in a gradient elution manner,and electron spray ionization source was adopted in positive and negative ion scanning.RESULTS Fifteen constituents showed good linear relationships within their own ranges(R2>0.990 0),whose average recoveries were 75.91%-112.13%with the RSDs of 3.06%-10.66%.CONCLUSION This simple,sensitive and efficient method can be used for the quality control of Zhuriheng Dropping Pills.

This study was purposed to investigate the practicality of matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) for detection of single nucleotide polymorphisms (SNP) on jak2 gene in multiple myeloma (MM) cells. The DNA fragment containing 2 SNPs of jak2 (C428T and C643T) was amplified using PCR and was purified. The purified product was used as the template for primer extension (PEX). The small products of allele-specific reaction were purified, the SNPs on jak2 gene of 5 patients with MM and 5 healthy persons were detected by MALDI-TOF MS. The results showed that the distribution of genotype C428T and C643T was not different between MM patients and healthy persons, both of which are homozygous T/T. In conclusion, the method based on MALDI-TOF MS and PEX technique for detecting SNP in jak2 gene is rapid, accurate and reliable method, and can be used in clinical practice.
Case-Control Studies ; DNA Primers ; Genotype ; Humans ; Janus Kinase 2 ; genetics ; Multiple Myeloma ; genetics ; Polymorphism, Single Nucleotide ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods

OBJECTIVETo establish a rat model of full-thickness skin defect to receive bone marrow mesenchymal stem cell transplantation for wound repair.

METHODSA full-thickness skin defect measuring 4 cmx4 cm in 36 F344 rats, which were divided into 3 groups with the wound covered with alloskin graft, acellular dermal matrix, or petrolatum gauze. In vitro cultured BMSCs in the 5th passage were transplanted into the skin defect, and the time of wound dressing dissociation and number of transplanted Brdu-positive cells in the wound were observed 14 days later.

RESULTSThe alloskin graft resulted in significantly longer time before dressing dissociation, with greater number of Brdu-positive cells in the wound than the other two wound dressings (P<0.001). The acellular dermal matrix showed better effect than petrolatum gauze in terms of the dressing dissociation time and the viable transplanted cell number in the wound.

CONCLUSIONAlloskin graft can be ideal for covering the wound surface to protect the transplanted BMSCs in rats.

Animals ; Bone Marrow Cells ; cytology ; Dermis ; transplantation ; Female ; Male ; Mesenchymal Stem Cell Transplantation ; Random Allocation ; Rats ; Rats, Inbred F344 ; Rats, Wistar ; Skin ; injuries ; Transplantation, Homologous ; Wound Healing ; physiology