Anemia ; chemically induced ; Hepatitis C, Chronic ; drug therapy ; genetics ; Humans ; Polymorphism, Genetic ; Pyrophosphatases ; genetics ; Ribavirin ; adverse effects ; therapeutic use

12E7 Antigen ; Antigens, CD ; metabolism ; Bone Neoplasms ; metabolism ; pathology ; surgery ; Cell Adhesion Molecules ; metabolism ; Chondrosarcoma, Mesenchymal ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Hemangiopericytoma ; pathology ; Humans ; Lung Neoplasms ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Pneumonectomy ; methods

Objective:To establish a real-time fluorescent quantitative PCR for the detection of torque teno virus types 7 (TTV7), 8 (TTV8) and 10 (TTV10) and analyze its performance in clinical sample detection.Methods:Specific primers were designed based on the gene sequences of TTV7, TTV8 and TTV10 in GenBank. Recombinant plasmids of pMD19-T-TTV7, pMD19-T-TTV8 and pMD19-T-TTV10 were constructed and used as positive standard control to establish a real-time fluorescent quantitative PCR based on FAM-Eclipse probe method. The specificity and sensitivity of the established method were evaluated. Moreover, it was validated in terms of clinical sample detection.Results:The standard curve equations of the real-time fluorescent quantitative PCR for detecting TTV7, TTV8 and TTV10 were y=-0.340 2 x+ 114.780 0 ( R2=0.998 8), y=-0.351 1 x+ 114.940 0 ( R2=0.995 3) and y=-0.348 9 x+ 115.020 0 ( R2=0.991 7), respectively, and there was no cross-reaction with other viruses. The detection sensitivity of the established method for TTV7, TTV8 and TTV10 were 108 copies/μl, 84 copies/μl and 98 copies/μl, and the positive detection rates in clinical pediatric serum samples were 10.9%, 2.1% and 4.3%, respectively. Conclusions:The established real-time fluorescent quantitative PCR for detection of TTV7, TTV8 and TTV10 was featured by strong specificity and high sensitivity, which could be used for rapid TTV detection in clinical serum samples.

Objective To investigate the effect of recombinant human bone morphogenetic protein 2 (rhBMP-2) on the osteogenic activities of human osteosarcoma cell line SaOS-2. Methods SaOS-2 cells were exposed to rhBMP-2 for 12,24,48 h at 0(control) ,2,20,200 μg/L, respectively. The mRNA expression of alkaline phosphatase(ALP) and bone gla(BCP) were detected by real time polymerase chain reaction. Results The mRNA expression of ALP and BGP of SaOS-2 cells increased gradually with rhBMP-2. The mRNA expression of ALP of the 20 μg/L group exposed for 48 h(1.60 ± 0.64), and the 200 μg/L group exposed for 12,48 h(1.70 ± 0.41, 1.80±0.19) were significantly higher than those of control (12 h: 0.80±0.25, 48 h: 0.74±0.21, allP<0.05). The mRNA expression of BGP of the 2 μg/L group exposed for 24 h(1.67 ± 0.33), the 20 μg/L group exposed for 12,24 h(2.42 ± 0.13,1.82 ± 0.14) and the 200 μg/L group exposed for 12,24 h(1.46 ± 0.11,1.24 ± 0.07) were significantly higher than those of control( 12 h: 1.01 ± 0.14, 24 h: 0.84 ± 0.12, all P< 0.05). Conclusions rhBMP-2 can promote the mRNA expression of ALP and BGP of SaOS-2 cells. They have a dose-response relationship, but represent a different dose-response effect.

Objective To observe the effects of recombinant human parathyroid hormone 1 to 34(referred to as hPTH) on the expression level of alkaline phosphatase(ALP) and bone gla protein(BCP) in human osteosarcoma cell line SaOS-2(referred to as SaOS-2 cells). Methods SaOS-2 cells were subcultured and treated with 1, 10 and 100 nmol/L hPTH for 12, 24 and 48 h. Total cellular RNA was extracted, cDNA was synthesized by reverse doses of hPTH, different duration of action, and their interaction on the expression level of ALP mRNA of SaOS-2 cells was significantly different(F = 29.32, 2.92, 7.64, all P < 0.05). The expression level of ALP mRNA(0.78 ± 0.43, 0.71 ± 0.05, 0.75 ± 0.19, 0.76 ± 0.14) of SaOS-2 cells after treatment with 0, 1, 10 and 100 nmol/L hPTH for 48 h was lower than those of treated for 12 h(1.01 ± 0.16, 1.37 ± 0.38, 1.49 ± 0.16, 2.52 ± 0.70, all P< 0.05) and 24 h (1.80 ± 0.47, 1.30 ± 0.36, 1.27 ± 0.17, 1.17 ± 0.11, all P< 0.05). The expression level of ALP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 12 hours was higher than that of the control(P < 0.05); the expression level of ALP mRNA of SaOS-2 cells after treatment with 1, 10 and 100 nmol/L hPTH for 24 h interaction on the expression level of BGP mRNA of SaOS-2 were significantly different (F = 8.26, 10.33, 5.51, all P< 0.05). The expression level of BGP mRNA(1.17 ± 0.28, 0.98 ± 0.08, 0.92 ± 0.17 and 0.84 ± 0.59) of SaOS2 cells after treatment with 0, 1, 10 and 100 nmol/L hPTH for 48 h was lower than those of treated for 12 h( 1.01 ± 0.14, 1.21 ± 0.18, 1.34 ± 0.30, 1.68 ± 0.62, all P< 0.05), and 24 h(1.71 ± 0.35, 1.41 ± 0.47, 1.28 ± 0.31 and 1.01 ± 0.18, all P < 0.05). The expression level of BGP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 12 h was higher than that of those groups treated with 0 and 1 nmol/L hPTH(all P< 0.05). The expression level of BGP mRNA of SaOS-2 cells after treatment with 10 and 100 nmol/L hPTH for 24 h and 48 h was lower than those of the control(all P < 0.05). The expression level of BGP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 24 hours was lower than that the group treated with 1 nmol/L hPTH(P < 0.05). Conclusions In vitro, hPTH significantly enhances osteogenic activities of human osteoblast in a short time, however, with prolonged stimulation time, osteogenic activity can show a downward trend.

OBJECTIVETo analyze the status and trend of injury deaths in Chinese people, and provide basic evidence for injury interventions.

METHODSData came from 2004-2005 the 3rd national retrospective sampling survey of death cause and covered 31 province-level regions and 160 surveillance spots in the interior of China, Total 142 660 482 person years were investigated. To describe the status of injury deaths, the crude death rate, years of potential life lost (YPLL), working years of potential life lost (WYPLL) and the standardized death rate were calculated. The population used for standardization was from census in 2000 and each five-year was counted as an age group; To analyze the trend of injury deaths, the constitution of the death causes were calculated based on the data of 1991-2000 national disease surveillance system which covered more than 10 000 000 population and 145 surveillance spots.

RESULTSThe total number of residents in survey districts died of injury between 2004 and 2005 was 87 753 (male 59 664, female 28 089, urban 23 308, rural 64 445); the crude death rate of injury in China 2004-2005 was 61.51/100 000 and accounting for 10.10% of all deaths; the standardized death rate was 58.45/100 000, ranking the fourth among the main cause of death for Chinese people. The YPLL of injury was 1579.61 person years per 100 000 and the WYPLL was 1721.41 person years per 100 000. The crude death rate of injury was 81.76/100 000 in male and 40.31/100 000 in female; the standardized death rates were 79.96/100 000 and 36.25/100 000, respectively. Injury mortality in male was two times higher than that in female. The crude death rates of injury were 48.66/100 000 in urban area and 68.01/100 000 in rural area; the standardized death rate were 44.08/100 000 and 66.25/100 000, respectively; the mortality in rural area was 1.4 times higher than that in urban area. The mortality for the aged 15 - 44 was 48.94/100 000(35 497/72 531 671) and accounting for 40% of all deaths, injury was the first cause of death for the aged 15 - 44. During 2004-2005, the top five causes of death related to injury were traffic accidents, suicide, falls, drowning and poisoning; the cases were 29 669, 18 678, 10 901, 7752, 4857 respectively in survey districts; the crude death rate were 20.80/100 000, 13.09/100 000, 7.64/100 000, 5.43/100 000, 3.40/100 000 respectively. From 1991 to 2005, the proportion of all injury deaths due to traffic accident increased from 15.00% (1551/10 338) to 33.79% (14 792/43 774) which showed a rising trend, the proportion of all injury deaths due to suicide decreased from 26.66% (2756/10 338) to 20.46% (8599/43 774) and the proportion of all injury deaths due to fall increased from 5.15% (532/10 338) to 12.87% (5630/43 774).

CONCLUSIONInjury is the primary cause of death resulting in premature death among Chinese people, traffic accident is the first cause of injury death. Since 1990s, the pattern of injury mortality of Chinese people has changed.

Accidents ; mortality ; statistics & numerical data ; Adolescent ; Adult ; Aged ; Cause of Death ; trends ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Young Adult

Objective To investigate the relationship between the oxidized low-density lipoprotein(ox-LDL) in the blood from the coronary sinus and impaired endothelium-dependent vasodilation in patients with coronary artery disease(CHD).Methods Four groups of patients were subjected,including acute myocardial infarction group(AMI,n=22),unstable angina group(UA,n=29),and stable angina group(SA, n=25) and control group(n=20) who was first suspected as CHD and then verified with normal coronary angiograms.Blood form the coronary sinus was collected through cardiac cathetering and the ox-LDL level was measured by Sandwich ELISA method.Brachial arterial hyperemia-induced flow mediated dilation(FMD) and sublingual nitroglycerin(NTG) mediated vasodilation were measured by high resolution ultrasound.Results The blood level of ox-LDL in patients with CHD was markedly higher than those in control group(P

OBJECTIVESTo evaluate the efficacy and toxicity of the B-NHL-BFM-90 protocol in the treatment of Chinese childhood and adolescent B-cell non-Hodgkin's lymphomas (B-NHL).

METHODSForty-two untreated childhood and adolescent B-NHL were enrolled in the present study. Of them 18 cases were Burkitt's lymphoma, 16 diffuse large B cell lymphoma and 8 anaplastic lymphoma. There were 10 cases in stage II and 32 in stage III/IV. The patients were grouped by risk factors into low, medium and high risk groups. All patients were treated with the B-NHL-BFM 90 (Berlin-Frankfurt- Münster) protocol. The low risk group received A, B courses for 4 cycles, the medium risk group AA, BB courses for 6 cycles, and the high risk group AA, BB, CC courses for 6 cycles.

RESULTSComplete remission (CR) was obtained in 37 patients (88%), and partial remission (PR) in 5 (12%). Of the 5 PR patients, I received autologous hematopoietic stem cell transplantation, 3 received radiotherapy for residual disease and 1 just under watching. Major toxicity was myelosuppression and mucositis, especially in AA, BB and CC cycles, but was tolerant and manageable. Median follow-up was 20 (4 - 89) months. Kaplan-Meier method was used to analyse survival data. Two year event free survival (EFS) for all patients was 86. 24%, being 100% for stage II and 80.95% for stage III/IV.

CONCLUSIONShort term and intensive chemotherapy can improves the efficacy and survival rate of childhood and adolescent B-NHL, especially for advanced stage patients.

Adolescent ; Adult ; Antineoplastic Combined Chemotherapy Protocols ; administration & dosage ; adverse effects ; Child ; Child, Preschool ; Feasibility Studies ; Female ; Follow-Up Studies ; Humans ; Infant ; Lymphoma, B-Cell ; drug therapy ; Male ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo study the differentiation potential of human umbilical cord-derived mesenchymal stem cells (UCMSC) into human sweat gland cells (hSGC) and the role of extracellular signal-regulated kinase (ERK) pathway.

METHODSUCMSC and hSGC were isolated and cultured in vitro. The former was identified with expression of CD14, CD29, CD34, CD44, CD45, CD105, cytokeratin 7 (CK7), CK19, and carcinoembryonic antigen (CEA), while the latter was identified with expression of CK19 and CEA. UCMSC with density of 5 x 10(4) cells per well placed in lower compartment of Transwell chamber were divided into control group (C, cultured with nutrient solution without any stimulation), thermal injury group (TI, treated with heat-shocked hSGC with density of 1 x 10(4) cells per well inoculated into the upper compartment of Transwell chamber for indirect co-culture), thermal injury + EGF group (TIE, treated with indirect co-culture as used in TI group, with addition of 50 ng/mL EGF), thermal injury + PD98059 group (TIP, treated with indirect co-culture as used in TI group, with addition of 10 nmol/mL ERK specific inhibitor PD98059) according to the random number table. One week after culture, the positive expression rates of CK7 and CK19 in UCMSC were detected by flow cytometry, the expression of CK19 and CEA in UCMSC were examined with immunohistochemical staining and the positive expression rate of CEA was calculated, and the expression level of phosphorylated ERK (pERK) was determined by Western blotting. Data were processed with one-way analysis of variance.

RESULTS(1) CD29, CD44, and CD105 were highly expressed in UCMSC, accompanied by low or negative expression of CD14, CD34, CD45, CK7, CK19, and CEA. The expression of CK19 and CEA were positive in hSGC. The two results showed that UCMSC and hSGC were pure. (2) Compared with those of C group [(2.2 +/- 1.5)%, (2.2 +/- 0.7)%, (3.3 +/- 0.7)%, 0.640 +/- 0.026], the expression levels of CK7, CK19, CEA, and pERK in UCSMC of TI group [(6.4 +/- 0.7)%, (5.7 +/- 0.3)%, (7.4 +/- 1.0)%, 0.790 +/- 0.049] and TIE group [(14.3 +/- 1.0)%, (12.6 +/- 1.1)%, (17.6 +/- 2.3)%, 1.200 +/- 0.032] were significantly increased (with F value respectively 78.49, 139.36, 87.13, and 191.74, P values all below 0.01), and those of TIE group were higher than those of TI group (with F value from 50.14 to 145.47, P values all below 0.01). There were no obvious difference in the 4 indexes between TIP group and C group (with F value from 0.00 to 0.13, P values all above 0.05).

CONCLUSIONSUCMSC co-cultured with heat-shocked hSGC can differentiate into hSGC, and ERK signal pathway participates in the process of differentiation of UCMSC into hSGC.

Cell Differentiation ; Cells, Cultured ; Extracellular Signal-Regulated MAP Kinases ; Flow Cytometry ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Signal Transduction ; Sweat Glands ; cytology ; metabolism ; Umbilical Cord ; cytology ; metabolism

ADAM Proteins ; biosynthesis ; genetics ; ADAMTS13 Protein ; Adult ; Aged ; Carcinoma, Hepatocellular ; metabolism ; Cell Line ; Female ; Humans ; Liver Neoplasms ; metabolism ; Middle Aged ; von Willebrand Factor ; biosynthesis ; genetics