Objective To study the clinic features and the mechanism of skin damage induced by high-dose methotrexate(HD-MTX).Method Children treated with HD-MTX were enrolled in a retrospective study in which the incidence and presentations of skin damage were concluded.Mechanism of the damage was studied base on skin pathology of an animal model.Result Skin damage presented in 3～9(median 5) days after HD-MTX and usually followed by bullation and exfoliation.Twelve cases,more severe than the others,accompanied with fever and symptoms of other organs especially the respiratory tract and the intestine.Skin damage was reduced when large dose rehydration used before,during and after HD-MTX.Conclusion HD-MTX may cause scald-like skin damage directly by its prolonged toxic effect on epidermal cells.Besides extended tetrahydrofolic acid rescue and supportive treatments,anti infection is the most important in dealing this condition.

Objective To explore the prognostic significance in monitoring minimal residual disease (MRD) in childhood acute lymphoblastic leukemia (ALL) by a simple method,and to detect cloned immunoglobulin H (IgH) and T cell receptor γ (TCRγ) gene rearrangements by using multiplex polymerase chain reaction (PCR) and automated fragment analysis.Methods Bone marrow samples were collected from 86 newly diagnosed cases of childhood ALL at the Department of Pediatrics,the First Affiliated Hospital of Sun Yat-Sen University,from May of 2009 to August of 2013.IgH and TCRγ gene rearrangements were amplified by qualitative multiplex PCR.The clonality of PCR production was analyzed by GENEMAPPERID software.Only those carried monoclonal IgH/TCRγ on diagnosis were arranged to monitor MRD.Detectable monoclonal IgH/TCRγby the end of induction was defined as MRD positive.All patients were treated with GD2008 ALL protocol.Clinical data of all newly-diagnosed ALL patients in the corresponding period were reviewed.The final follow-up on May 31,2014.Survival rates and event free survival (EFS) curves were estimated by the Kaplan-Meier,and compared by using the log-rank test.Results The percent age of 94.2 (81/86 cases) patients was at least 1 marker positive.Subsequent MRD was monitored in 79 cases.The median follow-up time was 20 months (9-61 months).By the end of induction,20 cases were MRD positive and 59 cases were M RD negative,and the 3-year EFS were 56.4％ ± 14.7％ and 94.0％ ± 3.4％ (x2 =8.563,P =0.003),respectively.According to the traditional prognostic stratification criteria,MRD was detected 65 cases in the non-high risk group:23 cases in standard risk group and 42 cases in intermediate risk group,and the difference of 3-year EFS had no statistical significance (95.3％ ±4.7％ vs 76.6％ ±9.0％,x2 =0.934,P =0.334).While using MRD by the end of induction as a risk stratification criterion,there was a statistical significant difference compared with the 3-year EFS for MRD-negative (n =52) group and MRD-positive (n =13) group (93.1％ ± 3.8％ and 59.5％ ± 16.2％,x2 =7.128,P =0.008).Conclusions It is a simple but feasible method to monitor MRD in childhood ALL by using this qualitative multiplex PCR with automated fragment analysis for monoclonal IgH/TCRγ gene rearrangements.MRD by the end of induction can be used as a more accurate risk stratification criterion than the traditional one.It is worth of further research.

Objective To evaluate the effect of delayed ligature of the umbilical cord on the blood volume in premature infants in early time. Methods From Oct 2007 to Jun 2010,194 premature infants with vaginal birth in department of genecology and obstetrics were randomly divided into routine ligature of the umbilical cord group (100 cases) and delayed ligature of the umbilical cord group (94 cases), comparision were made on ligature time, blood routine examination,blood pressure,oxyhemoglobin saturation and anemia incidence in 24 h,48 h,and 72 h after birth,and hemobilirubin on 1 d,2 d,3 d,and 7 d after birth. Results There were significant differences between the two groups in delayed ligature time [(100. 14 + 10. 57) s vs (22. 40 +5.02) s,P ＜0. 05)]. The hemoglobin and blood pressure in delayed ligature umbilical group were increased significantly(P ＜ 0. 05). There was 3 cases (3. 19%) of anemia in delayed ligature of the umbilical cord group, and 11 cases (11.00%) in routine ligature of the umbilical cord group, which showed significant difference (P ＜ 0. 05). And the hemobilirubin on 7 d after birth showed no significant difference between the two groups(P ＞ 0. 05). Conclusion Delayed ligature of umbilical cord has a significant influence on hemoglobin and blood pressure of premature infant. It can increase the blood volume, and reduce the incidence of anemia in early time.

Objective To examine the effect of labouring in water on analgesia of vaginal delivery.Methods From June 2009 to February 2011,38 women who had volunteered to deliver in water in the obstetrical department were set as the observation group,another 70 women who chose vaginal delivery were named as the control group in the corresponding period.The items of labor pain,satisfaction with the birth experience,length of labor and neonatal outcomes were compared.Results The pain level of the observation group after labouring in water decreased compared with that before immersion in water.The delivery course was not influenced with a high rate of vaginal delivery.Conclusions Labouring in water can alleviate delivery pain,increase rate of vaginal delivery with no influence on delivery course and outcome of mothers and infants.It is a safe and effective analgesia method which should be widely applied.

Objective To review the diagnosis of idiopathic pulmonary hemosiderosis ( IPH),and to evaluate the efficacy of maintenance therapy with dose-adjusted 6-mercaptopurine (6MP) in IPH children. Methods The diagnosis of IPH was confirmed by in-patient examination and at least 1 year follow-up to exclude secondary causes of pulmonary hemorrhage. Fifteen children met the criteria of IPH and were enrolled. The age at diagnosis was 2-13 years ( median 7 years). Prednisone was administered at 2 mg/( kg·d) for 4 weeks in acute phase of the disease followed by taper. 6MP was also started at 60 mg/( m2·d) simultaneously and continued for 3 years. Results The diagnosis was delayed in most children, which was due to the lack of initial classical manifestation of the disease. The time between the onset of symptoms and diagnosis ranged from 2 weeks to 108 months ( median 8 months) . All the patients exhibited response to the initial treatment and prednisone was successfully tapered off. Only 1 of 8 patients with relative leucopenia (3 × 109/L -6 × 109/L) on 6MP maintenance recurred while 5 of 7 others recurred (P < 0. 05) during median 6-year (range 2. 5 - 9. 5 years) follow-up. Of the latter 5 patients who recurred,4 remained recurrence-free after adjusting the dose of 6MP upwards to keep relative leucopenia. Conclusions Diagnostic delayed is still a main problem in pediatric IPH. Most IPH children in our group tolerated maintenance treatment with 6MP and achieved long-term remission, and these suggested growth retardation on long-term steroids therapy could be avoided. Because of interindividual difference in 6MP metabolism, adjusting the dose of 6MP may be necessary for treatment of IPH children and avoid under-treatment or overtreatment in some children,and thus improve the prognosis. White blood count could be a simple and useful indicator to predict clinical response in most IPH children on 6MP.

[Objective]To explore the effect and the possible mechanism of the proteasome inhibitor MG132 on acute T lympho?blastic leukemia cells.[Methods]The influence of different concentrations of MG132 in the viability and proliferation of CCRF-CEM was measured by MTS. Apoptosis rates of CCRF-CEM treated by MG132 were determined by flow cytometry. After being exposed to MG132,the protein levels of FOXO3a in cytoplasm and nucleus were analyzed by Western blotting. qRT-PCR was applied to detect the mRNA of FOXO3a and Puma in cells treated by MG132. Then CCRF-CEM was stably transfected with antisense FOXO3a using Lentivirus infection. We further investigated the effects of MG132 in FOXO3a-shRNA cells and elucidated the mechanisms of FOXO3a and Puma.[Results]MG132 inhibits the proliferation of CCRF-CEM,but has no cytotoxicity in peripheral blood mononu?clear cells(PBMC). Cellular apoptosis was induced in cells treated with MG132. At mRNA level,MG132 had no influence on FOXO3a,but increased the expression of Puma. However,MG132 promoted the expression of both FOXO3a and Puma at protein level. Interestingly,the expression of FOXO3a increased very little in cytoplasm. In FOXO3a-shRNA cells the expression of FOXO3a and Puma decreased at protein level. FOXO3a's knockdown attenuated the proliferation inhibition mediated by MG132.[Conclusion]MG132 inhibits the proliferation and promotes to apoptosis of CCRF-CEM. One of the mechanism is that MG132 inhib? its the degradation of FOXO3a,and then activates FOXO3a/Puma pathway.

AIM and METHOD: The adhesion behavior of bone marrow stromal cells(BMSC) in children with acute lymphoblastic leukemia(ALL) to bone marrow mononuclear cells(BMMC) and Raji cells were investigated by MTT method. The expression of adhesion molecules ICAM-1 and VCAM-1 were detected by flow cytometry. RESULTS: The adhesion ability of BMSC in acute period of ALL to BMMC was lower than that of control group. The adhesion ability of BMSC in long term remission period of ALL to Raji cells higher than that of control group. The expression of ICAM-1 on the surface of BMSC in acute period of ALL is much lower than that of control group. CONCLUSIONS: The adhesion of BMSC to BMMC or tumor cells in children with ALL was abnormal. The abnormal adhesion behavior might be partially due to the changed expression of adhesion molecules on BMSC.

Objective For further improving the prognosis of childhood acute promyelocytic leukemia (APL) in China,the treatment efficacies, outcomes and costs of protocols for childhood APL between in developed countries and in our hospital were compared. Methods 30 cases aged ＜15 years were diagnosed according to the FAB classification and detection of PML-RARα rearrangement. From December 1999 to September 2004,sixteen eligible children were treated with an intensive in-house protocol including high-dose Ara-C and anthracycline for post remission treatment. From September 2004 to January 2008,14 cases enrolled were treated with a less intensive protocol modified from the PETHEMA LPA99. Results The 3.5 years EFS was 37.5 % (s-x=0.121) for total 16 patients on in-house protocol. Six patients (37.5 %) abandoned treatment,2 died of intracranial hemorrhage at diagnosis (6.3 %) and sepsis in remission (6.3 %),respectively,and 2 relapsed (12.5 %). The 14 cases treated with modified PETHEMA had a 3.5 years EFS of 79.6 % (s-x=0.136). One died of intracranial hemorrhage at diagnosis (7.1 %) and 1 relapsed (7.1 %). Patients on modified PETHEMA had a significantly higher EFS (P=0.012),lower frequency of sepsis during treatment (7.7 % vs 77.8 %; P=0.0015) and lower hospitalization cost (median,RMB 35 200 vs 150 000; P ＜0.0001) than those on in-house protocol. Conclusion Treatment with the less intensive protocol based on the PETHEMA LPA99 study for childhood APL successively reduced complication of chemotherapy and reduced hospitalization cost without increasing relapses, which led to decreases in treatment-related toxicity and treatment abandonment rate,thus improving overall outcome.

[Objective] To evaluate the effect of fast rehabilitation nursing pathway in patients after artificial hip replacement.[Methods] 86 patients underwent artificial hip replacement were randomly divided into the control group and the experimental group with 43 cases in each group.The control group received routine care,while the experimental group was given fast rehabilitation nursing pathway.Anxiety and pain at 48h after operation were evaluated,and hip joint function(Harris score)was evaluated six months after operation.[Results] Compared with the control group,anxiety and pain score at 48h after operation decreased markedly,and Harris score six months after operation increased significantly.[Conclusions] Fast rehabilitation nursing pathway can reduce both the short-term and long-term efficacy of patients after artificial hip replacement.

Objective To explore the effect of Flavokawain B on the proliferation and apoptosis of acute T lymphoblastic leukemia(T -ALL)cells and its preliminary mechanism.Methods After the T -ALL cell lines CEM-C7(sensitive to glucocorticoids)and MOLT -4(resistant to glucocorticoids)cells were treated with different concentrations of Flavokawain B,the influence of Flavokawain B on the growth rate and doubling time of CEM-C7 and MOLT -4 cells was observed by 3 -(4,5 -dimethylthiazol -2 -yl)-5 -(3 -carboxymethoxyphenyl)-2 -(4 -sulfophenyl)-2H -tetrazolium(MTS)assay,and apoptosis was analyzed by using flow cytometry.Furthermore,Wes-tern blot assay was used to detect the expressions of Bim,Bcl -2 and cleaved Caspase -9.At last,the expressions of Bim and Bcl -2 in clinical T -ALL patient samples were also detected by using Western blot assay.Results MTS as-say showed that Flavokawain B significantly inhibited the cellular proliferation of T -ALL cell lines in a dose and time dependent manner(P <0.01 ).Flow cytometry findings revealed that Flavokawain B significantly induced the apoptosis of T -ALL cells in a dose -dependent manner(P <0.001 ).Western blot results indicated that Flavokawain B in-creased the expression of Bim and cleaved Caspase -9,and decreased the expression of Bcl -2 in T -ALL cell lines, which increased Bim and decreased Bcl -2 in clinical T -ALL patients samples,both in a dose -dependent manner. Conclusions Flavokawain B can inhibit the proliferation and induce the apoptosis of T -ALL cells by up -regulating the expression of Bim and down -regulating the expression of Bcl -2 and activating Caspase -9,whether resistant to glu-cocorticoids or not.