Objective To set a rapid,simple gene diagnosis method for Down syndrome.Methods Three short tandem repeats(D21S11,D21S1270,D21S1437)loci in or near Down syndrome critical region(DSCR) were analyzed and detected by polymerase chain reaction and DNA quantitative analysis in 11 core ancestry.Results There were four types by DNA quantitative analysis to different individuals at a short tandem repeats(STR) locus.In type one,a homozygote of one allelic gene was detected.In type two,a normal heterozygote of two allelic genes was found,the content or two DNA electrophoresis bands was approximately 1∶1.In type three,a Down syndrome patient of two allelic genes was discovered.The quantity of two electrophoresis bands was nearly 2∶1.In type four,the patient showed three DNA electrophoresis bands which the content was approximately 1∶1∶1.Conclusion A rapid gene diagnosis and prenatal diagnosis method for Down syndrome can be used for quantitative analysis of STR polymorphism loci.

Objective To verify a new revised method with low usage amount of arsenic trioxide for determining urinary iodine by As(Ⅲ)-Ce4+ catalytic spectrophotometry using ammonium persulfate digestion.Methods The standard curve linearity,sample detection limit,precision and accuracy of determining urinary iodine of this modified method were verified according to Determination Methods of Chemicals in Biological Materials.Results The linear correlative coefficients of the 0-300 μg/L range and 300-1200 μg/L range calibration curve were-0.9998--1.0000(n =6) and-0.9998--1.0000,respectively.The detection limit for iodine was 1.3 μg/L.The relative standard deviations were 1.5％ (1.1/71.3)-2.5％ (6.2/244.9) when measuring 3 urine samples with iodine concentration of 71.3-244.9 μg/L,and 0.6％(2.4/388.5)-1.7％(17.3/1018.0) when measuring 3 urine samples with iodine concentration of 388.5-1018.0 μg/L,respectively(n =6).The test results of the four urinary iodine national standard materials with iodine concentration of 73.0,206.0,556.0 and 883.0 μg,/L were all within the given value range and the average value relative deviation was 1.8％ (1.3/73.0),0.4％ (0.8/206.0),0.2％ (1.0/556.0) and-1.6％(-13.7/883.0),respectively (n =6).The average recovery was 98.8％ with a range of 93.2％ (186.3/200.0)-103.4％(51.7/50.0) when measuring 3 urine samples with iodine concentration of 64.6-144.9 μg/L and 3 urine samples with iodine concentration of 346.8-574.4 μg/L,respectively.Conclusions This new modified method greatly reduces the amount of waste containing arsenic,and can directly take urine samples with high iodine concentration to digest and determine without dilution.It is performed with good standard linear curve,better precision and high accuracy,and in line with the analysis of biological samples requirements.

OBJECTIVETo investigate morphological change of osteoblasts cultured on titanium plates with different microarchitecture structure when exposured to fluid shear stress.

METHODS14 dynes x cm(-2) fluid shear stress was applied on osteoblasts cultured on 3 different commercially pure titanium plates: Polished treatment (PT), sandblast (SB), sandblasting and acid-base (SB-AB) surfaces. Scanning electron microscope (SEM) was adopted to observe the morphological changes after 0.5, 4, 7.5 h time point respectively.

RESULTSMorphologically, no significant changes were observed after 0.5 h and few osteoblasts were seen after 7.5 h on all 3 type of different surfaces, and significant changes could only be observed after 4 h. Osteoblasts were elongated and rearranged along the flow way on different levels on PT surface. Shape of cells was altered, from long fusiform suspending over depressed areas into polygon stretching out many synapsises tightly attached to pits on SB-AB surface. Osteoblasts on SB surface displayed similar change as SB-AB surface, besides, some cells were elongated along the way of flow, stretching out threadlike synapsises attached to edges of pits.

CONCLUSIONMorphological change of osteoblast responding to fluid shear stress in physiological range depends on substrate microarchitecture and varies with the time of fluid shear stress application.

Microscopy, Electron, Scanning ; Osteoblasts ; Stress, Mechanical ; Titanium

OBJECTIVETo explore the correlation between the recurrence of cerebral infarction and aspirin resistance (AR)/Chinese medical (CM) constitutions.

METHODSTotally 413 cerebral infarction patients took Aspirin Enteric-coated Tablet (100 mg per day) while receiving routine therapy, 5 days at least in a week. They were followed-up for 12 months. Aspirin sensitivity (AS) was determined using turbidimetry. CM constitutions among patients with different AS were compared. Ratios of AR patients and AS patients of different CM constitutions in cerebral infarction recurrent patients were compared. Platelet membrane glycoproteins (GP) II b HPA-3 gene polymorphism was detected by polymerase chain reaction (PCR) method. Correlation between recurrence of cerebral infarction and AR, bb genotypes, CM constitutions times AS were analyzed by Logistic regression.

RESULTSTotally 11 patients dropped out, 101 (25.12%)with recurrent cerebral infarction and 301 (74.88%) without recurrent cerebral infarction. There were 152 (37.81%) AR patients and 250 (62.19%) AS patients. AR accounted for 26.6% (80/ 301) and AS accounted for 73.4% (221/301) in non-recurrent cerebral infarction patients. AR accounted for 71.3% (72/101) and AS accounted for 28.7% (29/101) in recurrent cerebral infarction patients. There was statistical difference in AR and AS ratios (χ2 = 64.287, P = 0.000). The proportion of yin deficiency constitution (YDC) was the largest [28.3% (43/152)] in AR patients. The proportion of blood stasis constitution (BSC) was the largest [23.6% (59/250)] in AS patients. There was statistical difference in CM constitutions between AR patients and AS patients (χ2 = 21.574, P < 0.01). The former 4 recurrent rates occurred in AR patients of YDC, BSC, damp-phlegm constitution (DPC), qi deficiency constitution (QDC). YDC occupied the first place [22.4% (34/152)]. The former 4 recurrent rates occurred in AS patients of BSC, QDC, DPC, damp-heat constitution (DHC). BSC occupied the first place [3.2% (2/250)]. Compared with non-recurrent cerebral infarction patients and AS patients, bb gene occurred most often, but aa gene and ab gene occurred obviously lesser in non-recurrent cerebral infarction patients and AR patients (χ2 = 20.171, χ2 = 55.139, P < 0.01). AR and bb gene were positively correlated with recurrent cerebral infarction (OR = 18.423, P = 0.000; OR = 1.304, P = 0.028). Body constitutions interacted with AS (OR = 0.707, P = 0.000).

CONCLUSIONSRecurrent cerebral infarction was closely related to AR and constitutional types. The recurrence rate was higher in AR patients of YDC. GP I b HPA-3 bb genotype might be a risk factor for AR and recurrent cerebral infarction.

Aspirin ; therapeutic use ; Body Constitution ; Cerebral Infarction ; Drug Resistance ; Humans ; Medicine, Chinese Traditional ; Neoplasms ; Recurrence ; Yin Deficiency

0.05).In addition,in different medication intervals and the same total dosage(200 mg),there was no difference in the number of patients who reached ASAS20,ASAS50 anti BASDAI50 in both groups.The changes of other parameters were not observed.Conclusion Two dosages and different medication interval of rhTNFR-Fc have similiar efficacy onset time and maintenee period.Mean- while,at the same total dosage,there is no signifieant difference in therapeutic effect in the two dosage groups. However,50 mg(1/7 d)regimen has better compliance than 25 mg(1/3 d).

Objective To investigate the role of CD4~+CD25~+ Treg cells on the pathogenesis of ankylos- ing spondylitis(AS), and to study the machanism of tumor necrosis factor(TNF)-?blocker on the treatment of AS by detecting the number of CD4~+D25~+ Treg cells before and after the treatment. Methods The diagno- sis of 10 AS patients was made based on the 1984 modified New York criteria. The patients received subcuta- neou injection of recombinant human tumor necrosis factor receptor-Fc fusion protein(rhTNFR-Fc)(etaner- cept)50 mg weekly for 8 weeks and 10 heathy subjects were enrolled for control. The mononuclear cells were isolated from peripheral blood in beth patients and controls. The number of CD4~+CD25~+ T cells and CD4~+ CD25~(high)T cells and the expression of CTLA-4. were detected by flow cytometry. Results The proportion of CD4~+CD25~+ T cells(24?19)% in total CD4~+ T lymphocytes of peripheral blood and CD4~+CD25~(high)T/CD4~+ T (6?6)% from AS patients before treated with rhTNFR-Fc was higher than that in healthy volunteers and AS patients after treatment(P

AIM To study the chemical constituents from the leaves of Cyclocarya paliurus (Batal.) lljinskaja.METHODS The ethyl acetate fraction of methanol extract from C.paliurus leaves was isolated and purified by silica,ODS and semi-preparative HPLC,then the structures of obtained compounds were identified by spectral data.RESULTS Eleven compounds were isolated and identified as (6S,7R,8R)-7α-[(β-glucopyranosyl) oxy] lyoniresinol (1),lyoniside (2),(-)-lyoniresinol 3 α-O-β-D-xylopyranoside (3),dihydrodehydrodiconiferyl alcohol 4'-O-β-D-glucoside (4),kaempferol-3-O-α-L-rhamnoside (5),kaempferol-3-O-α-L-(4"-Z-p-cumaroyl)-rhamnoside (6),naringenin (7),4'-hydroxywogonin (8),1-(3',4'-dihydroxyphenyl)-7-(4"-hydroxyphenyl)-4-hepten-3-one (9),grasshopper ketone (10),p-hydroxybenzoic acid (11).CONCLUSION Except for compounds 5,11,all the compounds are isolated from this plant for the first time.

OBJECTIVETo observe the expression of p38 MAPK and HSP 70 in CA3 and dentate gyrus (DG) regions of the hippocampus of rats induced by limb ischemic preconditioning (LIP).

METHODSNinety-six rats were randomly divided into sham and LIP groups. And the animals in the LIP group were further divided into LIP 6 h, LIP 12 h, LIP 1 d, LIP 2 d, LIP 3 d, LIP 4 d and LIP 5 d subgroups according to the time of reperfusion after LIP. Immunohistochemical staining and Western blot were used to observe the expression of p38 MAPK and HSP 70 in CA3 and DG regions of the hippocampus.

RESULTSThe results of the immunohistochemical staining and Western blot were consistent, which indicated that there were fluctuation in the p-p38 MAPK and HSP 70 expression in CA3 and DG regions after LIP compared with those of the sham group. The expression of p-p38 MAPK began to be up-regulated 1d after LIP and reached its peak at 3 d and lasted for 4 d after LIP. However, the expression of HSP 70 was significantly up-regulated 2 d after LIP compared to the sham group, reached its peak at 3 d and lasted until the 4 d after LIP.

CONCLUSIONLIP up-regulates the expression of p38 MAPK and HSP 70 in the CA3 and DG regions of the hippocampus of rats.

Animals ; CA3 Region, Hippocampal ; metabolism ; Dentate Gyrus ; metabolism ; Extremities ; blood supply ; HSP70 Heat-Shock Proteins ; metabolism ; Ischemic Preconditioning ; Male ; Rats ; Rats, Wistar ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo investigate the cellular effects of Pien Tze Huang (PZH) in the HT-29 human colon carcinoma cell line.

METHODSThe viability of HT-29 cells was determined by MTT assay. A fluorescence-activated cell sorting (FACS) analysis with annexin-V/propidium iodide (PI) and JC-1 staining were performed to determine cell apoptosis and the loss of mitochondrial membrane potential, respectively. Activation of caspase 3 was evaluated by a colorimetric assay. The mRNA expression levels of Bcl-2 and Bax were measured by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSPZH, in a dose- and time-dependent manner, reduced viability and induced apoptosis of HT-29 cells. Moreover, PZH treatment resulted in the collapse of the mitochondrial membrane potential, activation of caspase 3, and an increase in the Bax/Bcl-2 ratio.

CONCLUSIONPZH inhibits the growth of HT-29 cells by inducing cancer cell apoptosis via regulation of the Bcl-2 family and activation of caspase 3, which may, in part, explain its anticancer activity.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Colonic Neoplasms ; enzymology ; pathology ; Drug Screening Assays, Antitumor ; Drugs, Chinese Herbal ; pharmacology ; Enzyme Activation ; drug effects ; HT29 Cells ; Humans ; Membrane Potential, Mitochondrial ; drug effects ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

Mouse L1210 leukemia cell line is widely used as a model in the study of tumorigenesis, as well as the efficacy of chemotherapeutic drugs; however, like other suspension cell lines, the mouse L1210 cell line has lowest transfection efficiency, that many barriers exist to study about the structure, function, as well as metabolism in leukemia cells. This study was aimed to obtain higher transfection efficiency of L1210 cell line to facilitate scientific research. The transfection efficiencies of nucleofector and liposome in L1210 leukemia cells were detected by converted fluorescence microscopy and flow cytometry using EGFP (enhance green fluorescent protein); cell viability was observed by trypan blue exclusion test. The results showed that the transfection efficiency of nucleofector primarily through reporter gene pEGFP by Amaxa Nucleofector(TM) nuclear transfer apparatus was significantly higher than lipofectamine 2000 transfection, furthermore, in the same cell density (2 × 10(6)/ml) and plasmid content (10 µg), the transfection efficiency of nuclear transfer apparatus default mode A-20 was higher than that of other modes (S-18, T-20). Its survival rate was up to 50.5% after 24 hours. Cell viability of liposome transfection reached to 88% after 24 hours, but the transfection efficiency was lower (< 1%). It is concluded that the nuclear transfer apparatus A-20 transfected L1210 can reach higher transfection efficiency up to 61.6%, which is significantly higher than that of lipofectamine transfection. The survival rate is up to 50.5% well meeting the needs of scientific research. Higher transfection efficiency is helpful for in-depth research about the morphology, functions and pathogenesis in leukemia model L1210, and provides more searching space for the treatment of leukemia diseases.
Animals ; Cell Line, Tumor ; Cell Nucleus ; genetics ; Cell Survival ; genetics ; Genes, Reporter ; Green Fluorescent Proteins ; genetics ; Liposomes ; Mice ; Transfection ; methods