Objective To understand the carrying condition of the mutation gene of neonatal deafness in Huzhou City and explore the significance of the combination screening of hearing and deafness genes. Methods 2258 newborns who were born in Huzhou maternal and child health care hospital were screened for hearing and deafness and were followed up to the age of 3. Two kinds of complementarity and relativity of screening were analyzed. Early hearing screening was used by transient evoked otoacoustic emission (TEOAE) screening, hearing rescreening was used by TEOAE and autoauditory brainstem response (AABR) . Collected the neonatal umbilical cord blood and detected GJB2, SLC26A4, GJB3, mitochondrial 12SrRNA 4 genes 20 deafness mutations. Results Early hearing screening failed in 550 cases, with a failure rate of 24.36％. Detected in 118 cases of deafness gene carriers, total carrying rate was 5.23％. GJB2, SLC26A4, GJB3 and mitochondrial 12SrRNA gene mutation rate were 3.10％, 1.46％, 0.58％ and 0.09％ respectively. Initial failure rate of mother and child hearing screening was 16.30 ％ (300/1840) . The rate of gene transfer for deafness mutation was 5.65 ％ (104/1840) . Initial failure rate of NICU hearing screening was 59.81 ％ (250/418) . The rate of gene transfer for deafness mutation was 3.35％ (14/418) . The failure rate of initial hearing screening of NICU newborns was higher than that of mother and child (P<0.01) . There was no statistically significant difference in carrying rate between the two groups (P>0.05) . There was no statistical correlation between initial hearing screening andwhether or not to carry deaf mutation gene (P>0.05) . 52 infants were missed in this study. 12 patients were diagnosed with hearing impairment, and the hearing impairment rate was 0.54％. Among them, 9 cases were normal and 3 cases were abnormal. Conclusion Newborns hearing screening by whether or not had nothing to do with deafness gene.Hearing screening with deafness gene screening at the same time can reduce the delay in diagnosis of deafness and drug deafness can also be prevented early.

This study was aimed to explore the expression of MIP-1alpha, MCP-1 and their receptors CCR-1, CCR-2 in bcr/abl fusion gene positive CML cells, and to study the effects of P210(bcr/abl) fusion protein tyrosine kinase on expression of MIP-1alpha, MCP-1 and their receptors CCR-1, CCR-2 mRNAs in chronic myeloid leukemia cells. The expression levels of MIP-1alpha, MCP-1 and their receptors CCR-1, CCR-2 mRNA were detected by semi-quantitative RT-PCR in bcr/abl negative cells, bcr/abl positive cells, and P210(bcr/abl)-Rb-C-Box positive cells. The results showed that MIP-1alpha and CCR-1 mRNAs were expressed in bcr/abl negative cells, but not in positive cells. Both MCP-1 and CCR-2 mRNA cannot be detected in both bcr/abl positive and negative cells. After inhibiting P210(bcr/abl) tyrosine kinase activity by Rb-C-Box, expressions of MIP-1alpha and CCR-1 mRNAs were restored to normal (similar to P210(bcr/abl) negative cells). It is concluded that P210(bcr/abl) fusion protein inhibits the expression of MIP-1alpha and CCR-1 in chronic myeloid leukemia cells, but does not inhibit MCP-1 and CCR-2 mRNA expressions in these leukemia cells.
Chemokine CCL2 ; biosynthesis ; genetics ; Chemokine CCL3 ; Chemokine CCL4 ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; Macrophage Inflammatory Proteins ; biosynthesis ; genetics ; Receptors, CCR1 ; Receptors, CCR2 ; Receptors, Chemokine ; biosynthesis ; genetics ; Tumor Cells, Cultured

OBJECTIVESTo explore the effects of p210 bcr/abl fusion gene on expression of beta1 integrin and L-selectin mRNAs in mouse chronic myeloid leukemia (CML) cells.

METHODSComparisons of beta1 integrin and L-selectin mRNA levels among p210 bcr/abl negative, p210 bcr/abl positive, and p210 bcr/abl-Rb-C-Box positive cells were undertaken by quantity RT-PCR.

RESULTSIn p210 bcr/abl positive cells, L-selectin mRNA level was decreased, but beta1 integrin mRNA expression had no change as compared to those in p210 bcr/abl negative cells. When inhibition of bcr-abl tyrosine kinase activity by Rb-C-Box, the L-selectin mRNA expression restored to normal (similar to p210 bcr/abl negative cells).

CONCLUSIONp210 bcr/abl oncoprotein inhibits expression of L-selectin mRNA, but not of beta1 integrin mRNA.

Animals ; Cell Line, Tumor ; Fusion Proteins, bcr-abl ; genetics ; Gene Expression Regulation, Leukemic ; Genes, Retinoblastoma ; genetics ; Integrin beta1 ; genetics ; L-Selectin ; genetics ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Mice ; RNA, Messenger ; genetics ; Transfection

To explore the expression spectra of apoptosis-related gene pnas-2 in normal tissues and acute leukemia (AL) patient tissues, the expressions of pnas-2 gene in tissues including heart, brain, placenta, lung, liver, skeletal muscle, kidney, pancreas, spleen, lymph node, thymus, leukocyte, bone marrow and fetal liver were detected by Northern blot. The expressions of pnas-2 in samples including 44 de novo, 9 non-CR, 27 CR and 12 relapsed AL patients were measured by real-time RT-PCR and Northern blot, and the expression levels of pnas-2 in normal and tumor tissues from 31 patients with malignancies were also detected. The results showed that pnas-2 was not expressed in the most tissues except in placenta. The results of real-time PCR indicated that pnas-2 expressions in samples of de novo, non-CR and relapsed patients ware significantly higher than that in CR, tumor tissues and normal tissues. In serial monitoring of 7 AL patients, the expression level of pnas-2 was high at first visit examination, but remarkably decreased after remission, and the pnas-2 expression level increased again when relapsed. It is concluded that the pnas-2 is specifically up-regulated in acute leukemia patients, which might be an oncogene and participate in leukemogenesis.
Acute Disease ; Apoptosis ; genetics ; Apoptosis Regulatory Proteins ; genetics ; metabolism ; Biomarkers, Tumor ; genetics ; Gene Expression Regulation, Leukemic ; Humans ; Leukemia ; pathology

OBJECTIVETo investigate the expression of VEGF mRNA and secretion of VEGF protein in NB4 and HL-60 cells affected by all-trans retinoic acid (ATRA) and daunorubincin (DNR) respectively.

METHODSSemi-quantitative RT-PCR and ELISA were used to study the expression of VEGF mRNA and secretion of VEGF protein in NB4 and HL-60 cell lines treated by ATRA and DNR respectively.

RESULTSVEGF was expressed in both NB4 and HL-60 cells. The expression of VEGF mRNA and secretion of VEGF protein could be down-regulated by ATRA and DNR respectively in a time and dose dependent manner.

CONCLUSIONBesides inducing apoptosis and restraining proliferation of leukemic cells, ATRA and DNR exerted their anti-leukemia effects by reducing angiogenesis via reduction of angiogenic reaction stimulating signals.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Daunorubicin ; pharmacology ; Gene Expression Regulation, Leukemic ; drug effects ; HL-60 Cells ; Humans ; Leukemia ; drug therapy ; genetics ; metabolism ; physiopathology ; Tretinoin ; pharmacology ; Vascular Endothelial Growth Factors ; genetics ; metabolism

OBJECTIVETo assess the safety of dietary lead and cadmium intake in 3 areas of Zhejiang province.

METHODSUsing the total dietary study method, the study was conducted in 3 regions which represented coastal, city and rural areas in Zhejiang province from 2009 to 2010. The dietary survey was conducted on the residents (512 subjects) and the categories and volume of food consumption were obtained. The analytical food samples were obtained by food consumption survey, food aggregation, food sampling and preparation. The food samples were detected by inductively coupled plasma mass spectrometry (ICP-MS). The safety of dietary lead and cadmium intake was evaluated.

RESULTSThe median dietary lead intake (P₅₀) in Zhejiang province was 37.8 µg/d. The 97.5% dietary lead intake (P₉₇.₅) was 72.3 µg/d. The P₅₀ dietary lead intakes of different age and gender groups ranged from 23.2 to 44.2 µg/d. The P₉₇.₅ dietary lead intakes of different age and gender groups ranged from 34.2 to 88.1 µg/d. The P₅₀ dietary cadmium intake in Zhejiang province was 9.6 µg/d. The P₉₇.₅ dietary cadmium intake was 15.7 µg/d. The P₅₀ dietary cadmium intakes of different age and gender groups ranged from 6.4 to 11.4 µg/d, accounting 15.6% - 42.6% of PTMI (provisional tolerable monthly intake, 25 µg/kg). The P₉₇.₅ dietary cadmium intakes of different age and gender groups ranged from 10.5 to 21.4 µg/d, accounting 27.5% - 77.6% of PTMI. Vegetable (11.3 µg), cereal (11.0 µg) and meat (9.8 µg) were the first three food sources which accounted for 84.9% of dietary lead (P₅₀: 37.8 µg/d). Cereal (3.6 µg), vegetable (2.1 µg) and legume (0.9 µg) were the first three food sources which accounted for 68.8% of dietary cadmium (P₅₀: 9.6 µg/d).

CONCLUSIONDietary lead and cadmium intakes of most residents in 3 areas of Zhejiang province as well as the average level are safe.

Adolescent ; Adult ; Aged ; Cadmium ; analysis ; China ; Diet Surveys ; Female ; Food Contamination ; analysis ; Humans ; Lead ; analysis ; Male ; Middle Aged ; Young Adult

OBJECTIVETo evaluate levels of lead (Pb) and cadmium (Cd) in the breast milk in the second postpartum month, to investigate the relationship between Pb/Cd levels in breast milk and some sociodemographic parameters, and to explore whether these levels affect the infants' physical status or the mothers' psychological status (postpartum depression).

METHODSA cross-sectional study was conducted between November 2009 and December 2010. Altogether 170 healthy mothers were enrolled from Nanjing Maternity and Child Health Care Hospital. The inclusion criteria were: voluntary to participate in this study, healthy, with no chronic disease, breastfeeding in the second postpartum month, living in a suburban but not non-industrial area of Nanjing, and not occupationally exposed to toxic metals. All the mothers completed a questionnaire and were evaluated based on the Edinburgh Postpartum Depression Scale (EPDS) to identify the risk of postpartum depression. Pb and Cd levels in breast milk were determined by inductively coupled plasma mass spectroscopy. The infants of these mothers were examined for their z scores of weight for age, length for age, head circumference for age, and body mass index for age.

RESULTSThe median breast milk levels of Pb and Cd were 40.6 μg/L and 0.67 μg/L, respectively. In 164 (96.5%) of the 170 samples, Pb levels were higher than the limit reported by the World Health Organization (> 5 μg/L). Breast milk Cd level was > 1 μg/L in 54 (31.8%) mothers. The mothers with a history of anemia had a higher breast milk Pb level than those without a history of anemia (41.1 μg/L vs. 37.9 μg/L, P = 0.050). The median breast milk Cd level in those who were active and passive smokers during pregnancy was significantly higher than that in non-smokers (0.88 μg/L vs. 0.00 μg/L, P = 0.025). The breast milk Cd level in the mothers not taking iron and vitamin supplements for 2 months postpartum was higher than in those taking the supplements (iron supplement: 0.74 μg/L vs. 0.00 μg/L, P = 0.025; vitamin supplement: 0.78 μg/L vs. 0.00 μg/L, P = 0.005). Breast milk Cd level at the second postpartum month was negatively correlated with the z scores of head circumference (r = - 0.248, P = 0.042) and weight for age at birth (r =- 0.241, P = 0.024) in girls. No correlation was found between the breast milk Pb/Cd levels and the EPDS scores.

CONCLUSIONConsidering the high levels of Pb and Cd in breast milk in this study, breast milk monitoring programs are necessary.

Adolescent ; Adult ; Cadmium ; analysis ; China ; Cross-Sectional Studies ; Female ; Humans ; Lead ; analysis ; Milk, Human ; chemistry ; Pregnancy

The latent membrane protein 2 (LMP2) is a kind of protein expressed by EBV-infected cells. This study was aimed to investigate whether the stimulation of peripheral blood mononuclear cells with peptides induces EBV-specific cytotoxic T lymphocytes (CTL). The peptides were mixture of LMP2 protein and available for people with different HLA types. Peripheral blood sample was collected from a patient with EBV-associated hemophagocytic syndrome. The mononuclear cells were isolated and cultured to obtain dendritic cells (DCs). Immature DCs were pulsed with MIX-LMP2 and added with different maturation-promoting factors. The auto-T lymphocytes were stimulated weekly with the harvested mature DCs loaded with MIX-LMP2, and totally for two times. Part of isolated lymphocytes was cultured without any stimulation as control. T-cell receptor (TCR) beta spectratyping was used to analyze the distribution of different T cell subgroups before and after culture. The phenotype of T lymphocytes was determined by flow cytometry. The IFN-gamma assay was used to estimate specific cytoxic activity of the cultured T cells. The results showed that the distribution of TCRbeta was changed according to analysis of TCR spectratypes. From the distribution of gene families of TCRbeta, the T lymphocytes were oligoclonal before culture, but shifted to a polyclonal after culture in vitro like the normalization of TCR diversity, suggesting the subgroups of lymphocyte could return to normal. The percentage of CD3+, CD3+CD8+ CD3+ CD45RA- CD45 RO+ on T lymphocytes from freshly isolated mononuclear cells were 70.73%, 42.99%, 27.56% respectively. After being stimulated twice with DC loaded with MIX-LMP2, they further increased to 95.17%, 52.54% and 81.41%. The percentages of CD3-CD56+ NK cells and CD4+CD35+ FOXP3+ regulation T cells seldom changed, from 2.12%, 0.03% to 2.35%, 0.02% respectively. The increase of CD3+CD45RA-CD45RO+ cells obviously indicated that most naive T cells could be activated. ELISA for IFN-gamma showed that when DCs loaded with LMP2 peptide were used as target cells, IFN-gamma level secreted by the T cells stimulated with LMP2 peptide-pulsed DCs was 805+/-16 pg/ml and 1729+/-49 pg/ml, the IFN-gamma level secreted by T cells stimulated twice with LMP2 peptide-pulsed DCs was 956+/-23 pg/ml and 2325+/-58 pg/ml respectively at effector-target ratios of 10:1 and 10:2. They were both significantly higher than that secreted by T cells without any stimulation (441+/-27 pg/m and 557+/-19 pg/ml) (p<0.05). But DCs unpulsed with LMP2 peptide were used as target cells, there were no significant differences between the T cells stimulated with LMP2 peptide-pulsed DCs and the T cells without stimulation (p>0.05). It is concluded that the antigen specific T cells recognizing EBV epitopes can be obtained by using DCs pulsed with MIX-LMP2 peptide in vitro, meanwhile the distribution of T cell subgroups can be changed and normalized.
Antigens, Viral ; immunology ; Cells, Cultured ; Coculture Techniques ; Cysteine Endopeptidases ; immunology ; metabolism ; Dendritic Cells ; immunology ; metabolism ; Epitopes, T-Lymphocyte ; immunology ; Epstein-Barr Virus Infections ; immunology ; Herpesvirus 4, Human ; immunology ; Humans ; Leukocytes, Mononuclear ; cytology ; T-Lymphocytes ; cytology ; immunology

OBJECTIVETo study the predictable value of monitoring minimal residual disease (MRD) regularly by flow cytometry (FCM) in patients with acute leukemia (AL) in the first complete remission (CR(1)).

METHODSFrom April 2005 to July 2009, AL patients who had got CR(1) after chemotherapy were regularly monitored for MRD in bone marrow by FCM to relapse or to July 2010 in Beijing Daopei Hospital (not including those received stem cell transplantation). The special antibody combinations were employed for each patient according to aberrant expression of leukemia cells. MRD(+) was defined as the aberrant cells more than 0.01%. The probability of continuous CR (CCR) was calculated by Kaplan-Meier formula, and the statistical difference between two CCR probabilities was evaluated by log-rank test.

RESULTSA total of 163 AL patients in CR(1) were monitored to relapse or to July 2010. Among 89 AML patients referred to our hospital within 1 year after diagnosis, 30 cases were in MRD(+) and 59 cases MRD(-) till 12 months following chemotherapy, 3/30 patients in MRD(+) and 47/59 remained in CCR to July 2010. The probability of CCR at 24, 36 months was 13%, 13%in MRD(+) group, 94%, 78% in MRD(-) group respectively, the difference between them was statistically significant (P < 0.01). Among 35 ALL referred to our hospital within 5 months after diagnosis, 13 cases were MRD(+) and 22 cases MRD(-) till 5 months following chemotherapy, 0/13 patients in MRD(+) and 20/22 patients in MRD(-) remained in CCR to July 2010. The probability of CCR at 24, 36 months was 0% in MRD(+) group, 96%, 96% in MRD(-) group respectively, the difference between them was statistically significant (P < 0.01). Over the time point above, all patients with MRD(+) or their MRD from negative to positive relapsed finally, and most patients with MRD(-) remained CCR to July 2010.

CONCLUSIONIt had a clinical prognostic value to monitor MRD regularly by FCM in the patients with AL after CR(1).

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Flow Cytometry ; Humans ; Leukemia, Myeloid, Acute ; diagnosis ; pathology ; Middle Aged ; Neoplasm, Residual ; diagnosis ; Predictive Value of Tests ; Prognosis ; Recurrence ; Young Adult

OBJECTIVETo investigate the effects of intensive insulin therapy on plasma nitric oxide (NO) and endothelin-1 (ET-1) levels in patients undergoing cardiac valve replacement under cardiopulmonary bypass (CPB).

METHODSA total of 36 patients were randomly assigned to routine therapy (RT) group and intensive insulin therapy (IT) group, with 18 patients in each group. The blood glucose levels during surgery were maintained at 3.9 to 10.0 mmol/L and those after surgery at 3.9 to 6.1 mmol/L in IT group, whereas patients in RT group didn't undergo the treatment of controlling glucose levels during operation and maintained below 13.9 mmoVL after operation. Levels of plasma NO and ET-1 in both groups were respectively measured before surgical anesthesia, at the initiation of CPB, and 0 h, 4 h, 12 h, 24 h and 48 h after the termination of CPB.

RESULTSIn RT group, plasma NO concentration was decreased since the initiation of CPB [from (68.2 +/- 16.3) micromol/L to (67.8 +/- 8.4) micromol/L] and reached the trough at the termination of CPB [ (60.0 +/- 10.2) micromol/L, P < 0.05 compared with that before anesthesia]. Then it began to increase and neared to the preoperational level 48 h after the termination of CPB. In contrast, plasma ET-1 concentration was increased since the initiation of CPB [from (62.2 +/- 10.2) ng/L to (68.3 +/- 10.8) ng/L] and reached the peak at the termination of CPB [ (112.5 +/- 18.6) ng/L, P < 0.01 compared with that before anesthesia]. Then it began to decrease and reached the preoperational level 24 h after the termination of CPB. In IT group, however, the changes of NO and ET-1 levels at different time points during CPB and thereafter didn't reach the significance as compared with those before anesthesia.

CONCLUSIONSIntensive insulin therapy may relieve the changes of CPB-induced NO and ET-1 levels during cardiovascular surgery, which suggests its protective effects on cardiovascular function.

Adult ; Cardiopulmonary Bypass ; adverse effects ; Endothelin-1 ; blood ; Female ; Humans ; Hyperglycemia ; drug therapy ; etiology ; Hypoglycemic Agents ; therapeutic use ; Insulin ; therapeutic use ; Insulin Infusion Systems ; Male ; Middle Aged ; Nitric Oxide ; blood