Loss of ATM activity induces GADD45α-dependent apoptosis in cerebellar granular neurons / 中山大学学报(医学科学版)

objectiveTo explore the specific molecular mechanism of neuronal apoptosis induced by ATM inactivation. MethodsCGNs matured 7 days in vitro were cultured 8 h with 25 K， 5 K or 25 K medium containing ATM-specific inhibitors （Ku55933， 10 µmol/L； Ku60019， 15 µmol/L） for Hoechst stain and apoptosis analysis， or cultured for different lengths of time （2， 4， 8 h） to detect the protein expression levels of ATM， caspase-3 and cleaved caspase-3 by Western blotting. ATM and GADD45α specific siRNA was transfected into C6 cells and CGNs， and its interference efficiency was verified by q-PCR and Western blotting. CGNs matured for 5 days in vitro were transfected with ATM specific siRNA and pCMV-EGFP by calcium phosphate for 48 h， Hoechst staining and apoptosis analysis were performed. CGNs matured for 7 days in vitro were treated with 25 K medium containing ATM specific inhibitors for 8 h， transcriptome sequencing， differential expression gene identification and pathway enrichment analysis were performed. CGNs matured for 5 days in vitro were co-transfected with GADD45α specific siRNA and pCMV-EGFP by calcium phosphate for 48 h， then treated with 5 K or 25 K medium containing 15 µmol/L Ku6 for 8 h. Hoechst staining and apoptosis analysis were performed. ResultsCompared with the 25 K， CGNs nuclear pyknosis rate， cleaved Caspase-3 and ATM protein expression level were increased in the 5 K and ATM-specific inhibitor groups. The mRNA and protein expression levels of ATM and GADD45α were effectively reduced after transfection of ATM and GADD45α specific siRNA in C6 cells and CGNs. Compared with control， CGNs transfected with ATM specific siRNA showed a higher nuclear pyknosis rate. Totally 835 genes were identified to be up-regulated and 848 genes to be down-regulated in the Ku55933 treatment group； 454 genes were identified to be up-regulated and 314 genes to be down-regulated in the Ku6 treatment group； 274 genes were co-up regulated in the Ku5 and Ku60019 treatment groups， while 179 genes were co-down-regulated in the Ku5 and Ku6 treatment groups and the expression of ATM downstream target GADD45α was upregulated. The enrichment results showed that TNF signaling pathway， NF-κB signaling pathway and Apoptosis signaling pathway were significantly enriched. Compared with control， mRNA and protein expression levels of GADD45α were increased in inhibitor treatment and 5 K， while knocking down GADD45α resulted in a decrease in nuclear pyknosis rate in the Ku60019 and 5 K treatment group. ConclusionLoss of ATM activity induces GADD45α-dependent cerebellar granular neuronal apoptosis.

Predictive value of umbilical cord blood bilirubin level for subsequent neonatal jaundice / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the predictive value of umbilical cord serum (UCS) bilirubin for subsequent jaundice in healthy term newborns.</p><p><b>METHODS</b>Five hundred and twenty-three healthy term newborns (275 boys, 248 girls) were selected. The cord blood total serum bilirubin concentration and the serum albumin concentration were determined. All the infants were assessed for jaundice daily by measurement of transcutaneous bilirubin (TCB). When the infant's TCB was >or= 18 within the first 24 h after birth, >or= 21 at 48 h, >or= 25 at or after 72 h, the venous total serum bilirubin (TSB) was determined and treatment against jaundice was applied as needed. The infants were aligned into four groups according to their UCS bilirubin levels, starting from < 30 micromol/L(group 1); >or= 30 micromol/L(group 2); >or= 36 micromol/L(group 3); >or= 42 micromol/L(group 4). The frequency of hyperbilirubinemia and phototherapy (PT) were compared among the four groups. An analysis of UCS bilirubin as a predictor of later development of jaundice was performed. The characteristics of the infants who became jaundiced (jaundiced group) were compared with the normal infants (non-jaundiced group).</p><p><b>RESULTS</b>A clear correlation between UCS bilirubin level and the development of hyperbilirubinemia was found in all populations of the four groups. Only eight of the 194 infants in group 1 showed a TCB index >or= 25. TSB values > 205 micromol/L but < 257 micromol/L were observed in 2 newborns. None of the infants in this group showed TSB > 257 micromol/L or needed PT. Thirty-two infants in group 2 showed TCB >or= 25, 12 infants had TSB > 205 micromol/L but < 257 micromol/L, 2 infants had TSB > 205 micromol/L and received PT. In group 3, one infant developed hyperbilirubinemia at 48 h after birth and received PT. Thirty-nine infants showed TCB >or= 25, 16 infants TSB > 205 micromol/L but < 257 micromol/L, 2 infants had TSB > 205 micromol/L and also received PT. In group 4, 4 infants showed a range of TSB from 200 to 215 micromol/L at 48 h and received PT. Twenty-two infants showed TCB >or= 25, 17 of them showed TSB > 205 micromol/L but < 257 micromol/L, and 5 of them had TSB > 205 micromol/L and received PT. The frequency of TSB > 205 micromol/L increased from 1.03% in group 1, 5.77% in group 2, 19.75% in group 3 and to 42.5% in group 4. None of the 194 newborns in group 1 needed phototherapy, whereas 0.96%, 3.70% and 22.5% of the newborns in groups 2 - 4, needed PT. The frequency of patients with hyperbilirubinemia or phototherapy increased with increasing UCS bilirubin levels. For the prediction of TCB >or= 25 using a UCS bilirubin cut-off level, such as >or= 35 micromol/L, we found a positive predictive value of 45.68% and sensitivity of 68.27%. It is significant to predict neonatal jaundice by UCS bilirubin levels (P < 0.001). In the jaundiced group (TCB >or= 25) UCS bilirubin levels were significantly higher than those in the non-jaundiced group (t = 10.96, P < 0.001). No significant differences were found in the cord blood serum albumin concentration (t = 2.38, P > 0.05), the gestational age (t = -0.90, P > 0.05), and birthweight (t = 0.10, P > 0.05) between the jaundiced and non-jaundiced groups.</p><p><b>CONCLUSIONS</b>UCS bilirubin level is useful in predicting the subsequent jaundice in healthy term infants. The use of UCS bilirubin values may help detect infants at low or high risk for hyperbilirubinemia and minimize an unnecessary prolongation of hospitalization.</p>

Treatment of advanced Wilms' tumor / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To improve prognosis of the patients with advanced Wilms' tumor, the authors compared different therapeutic strategies including preoperative transcatheter arterial chemoembolization (TACE), conventional preoperative chemotherapy and initial surgery.</p><p><b>METHODS</b>Sixty-two patients aged from 5 months to 10 years (mean 3.2 years) were identified from medical records to have histologically confirmed advanced Wilms' tumor during the period from January 1993 to December 2002. The criteria for choice were huge tumor size with a volume more than 550 ml or the mass extending beyond the midline, involvement of vital structures, inferior vena cava invasion, distal metastasis or bilateral Wilms' tumor judged by imaging examination. All cases were divided into 3 groups according to the treatment received: 31 cases in group TACE received preoperative transcatheter arterial chemoembolization with Lipiodol-Epirubicin (EPI)-Vincristine emulsion. One week after TACE, systemic chemotherapy with Actinomycin D (ACTD) was administered and tumor resected at two weeks after TACE. 20 cases in group PC received conventional preoperative chemotherapy with VCR, ACTD plus EPI for 4-5 weeks, and 11 cases in group IS underwent initial surgery. Postoperative treatment for all patients was based on the postoperative staging and tumor histology.</p><p><b>RESULTS</b>In the patients treated with TACE, no drug-induced complications such as cardiotoxicity, nephrotoxicity, hepatic dysfunction or bone marrow suppression were observed except for mild fever due to tumor necrosis. The percentages of tumor size shrinkage were 32.4% and 20.3% in group TACE and group PC, respectively (P < 0.05). Complete surgical removal of the tumor was achieved in 27 patients (87.1%) in group TACE, significantly higher in comparison with 14 in group PC (70.0%, P < 0.05) and 2 in group IS (18.2%, P < 0.01). Event-free survival (EFS) at 2 years was 87.1% (27/ 31), 60.0% (12/20) and 18.2% (2/11), respectivrely. EFS at 4 years was 84.6% (11/13), 56.3% (9/16 ) and 18.2% (2/11) in groups TACE, PC and IS, respectively.</p><p><b>CONCLUSION</b>The present study has shown that both preoperative TACE and conventional preoperative chemotherapy can be applied to the patients with advanced Wilms' tumor who are not candidates for immediately surgical resection. The survival is significantly increased in the patients undergoing preoperativeTACE when compared with conventional preoperative chemotherapy and initial surgery.</p>

Follow-up study of mental developments in high-risk children / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To study the mental developments in high risk children and the impact of the high risk factors on neurologic abnormalities, mental defect and long-term outcome.</p><p><b>METHODS</b>The mental development of 122 children who had been exposed to high-risk factors and treated between March 1994 to May 1995 during their newborn periods was evaluated. Gesell development scales were performed when they were at 6 and 12 months old. And Wechsler intelligence scales for children (Chinese version) were performed at 6 approximately 7 years old.</p><p><b>RESULTS</b>The children exposed to hypoglycemia during their newborn period and preterm labor had significantly lower IQ, VIQ and PIQ scores (P <0.05). The other risk factors in order were low birth weight, severe anoxia, asphyxia at birth, erythrocythemia, hyperbilirubinemia. There was significant difference between the children exposed to one risk factor and those exposed to two or more risk factors (P <0.05). And there was significant correlation between developmental assessment at 6 and 12 months and mental development at 6 approximately 7 years old (P<0.01).</p><p><b>CONCLUSION</b>The impact of the high risk factors at birth on children's mental development is not negligible. And the risk of development abnormalities will increase if the children were exposed to multiple risk factors. The evaluation of development at 6 approximately 12 months is of predictive value for long-term outcome.</p>