Abstract:Neonatal hypoxic2ischemic encephalopathy (HIE) is a severe complication of perinatal asphyxia. There are differences in the diagnostic criteria for HIE between China and overseas. The criteria of China do not include blood gas results of umbilical artery and incidence of multiple organs disorder (MOD) and can easily lead to misdiagnosis , while the oversea criteria are more scientific , perfect but over2strict which may result in missed diagnosis. At present , there are no highly specific and sensitive indexes for its diagnosis. So it is suggested that a diagnosis of HIE should be made according to history ,Apgar score , blood gas results , incidence of MOD ,brain images and clinic manifestations , etc.

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Objective It has been reported that nitric oxide (NO) might play an important role in the development of intrauterine growth retardation (IUGR). This paper aims at investigating the effect of L arginine (L Arg) on NO levels of maternal and neonatal umbilical blood in IUGR. Methods Sixty six pregnant women with IUGR were recruited. Thirty six cases were given routine therapy (Routine therapy group), and the other 30 cases were given L Arg combined with routine therapy (L Arg group). Another 30 cases with normal pregnancy were used as Normal control group. The NO levels of maternal and neonatal umbilical blood were monitored before and after treatment. Results After the treatment, the maternal serum NO level in the L Arg group was significantly higher than that of the Routine therapy group ( 58.42 ± 23.12 μmol/L vs 43.49 ± 20.27 μmol/L) (P< 0.01 ). The umbilical blood NO level in the L Arg group was alao significantly higher than that of the Routine therapy group ( 25.23 ± 12.05 μmol/L vs 16.95 ± 11.19 μmol/L) (P< 0.01 ). Conclusion L Arg can increase serum NO levels of both maternal and umbilical blood in IUGR.

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Objective Neonatal asphyxia may induce acute gastrointestinal injuries such as erosion, ulcer, bleeding and so on. Recent investigations have shown that epidermal growth factor (EGF) and its receptor (EGFR) play an important role while participating in the reparation of acute gastrointestinal injury in adult animals. This paper aims at studying the expressions of EGF and EGFR in gastric mucosa of neonatal rats with intrauterine asphyxia as well as their roles in the reparation of gastric mucosa lesions. Methods Wistar rats, 21 days pregnant, were used to establish animal models of intrauterine asphyxia. The surviring pups of the Asphyxia group and the pups without asphyxia (Control group) were sacrificed by decapitation at 0, 24, 48, 72 hrs after birth and gaster specimens were collected. The expressions of EGF, EGFR and EGFR mRNA in the gastric mucosas were assayed by immunohistochemistry and reverse transcription polymerase chain reaction (RT PCR) and the histology of gastric mucosas were observed. Results (1) The gastric mucosa lesions of neonatal rats were found in the Asphyxia group and became more and more serious untill a peak was reached 48 hrs after birth. (2) Low expressions of EGF and EGFR could be detected in the gastric mucosas of rats in the Control group, and there were no differences among various time points. (3) The expressions of EGF and EGFR in the gastric mucosas of neonatal rats in the Asphyxia group increased with time and reached the peak 48 hrs after birth. The EGFR expressions at 24, 48 and 72 hrs in the Asphyxia group were significantly higher than those of the Control group (P< 0.01 or 0.05 ). (4) The expression of EGFR mRNA in the gastric mucosa of neonatal rats in the Asphyxia group reached a peak 24 hrs after birth and was higher than that of the Control group (P< 0.01 ). Conclusions EGF and EGFR may play an important role in the reparation of gastric mucosa lesions of neonatal rats with intrauterine asphyxia.

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Objective The advanced solid tumor in children is not responsive to regular chemotherapy. This paper aims at studying the feasibility and effect of high dose chemotherapy combined with autologous peripheral stem cell transplantation (APBSCT) in the treatment of high risk advanced solid tumors in children. Methods Thirteen patients, including 7 cases of malignant lymphoma and 6 cases of neuroblastoma, were given APBSCT after receiving complete remission (CR) or partial remission (PC). The median disease duration before transplantation was 10 months. Before transplantation, 12 patients had CR and 1 had PR. Mobilization of stem cells was performed with chemotherapy plus G CSF or GM CSF in 11 and chemotherapy alone in 2 patients. The collecting mean number of MNCs, CD34 + cell and CFU GM was ( 6.85 ± 2.65 )×10 8/kg, ( 15.82 ± 12.93 )×10 6/kg and 17.87 ± 17.94 colons/10 4 cell respectively. The conditioning regimen consisted of cyclophosphamide plus TBI as basic program in 6 patients, melphalan plus etoposide, carboplatin in 5 patients and busulfan plus melphalan in 2 patients. Results The median times of the amount of WBC being more than 0.5 ×10 9/L and 1.0 ×10 9/L and of thrombocytopenia being more than 20×10 9/L were 12, 15 and 19 days respectively. A follow up of 48 months (ranging from 1 month to 144 months) showed that survival and death rates after transplantation were 77% (10/13) and 23% (3/13) respectively. No death due to the transplantation was found. Conclusion APBSCT can significantly improve the prognosis of patients with advanced solid tumors.

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Objective The therapeutic effect of chronic myeloid leukemia (CML) is undesirable. In order to find a new sensitive drug for CML, this study aims at exploring the effects of tetra arsenic tetra sulfide (As 4S 4) on human leukemia K562 cells. Methods The viability of K562 cells, represented by absorbance, was measured by MTS assay. The cells morphological changes were determined by Wright's staining and Hoechst33342 assay. The cell apoptosis was evaluated by DNA agarose gel electrophoresis and the cell apoptosis rate was measured by flow cytometry. Results The cell viability decreased significantly being cultured with 5.0 μmol/L As 4S 4, STI571 and herbimycin for 72 hrs (with absorbances of 0.32 ± 0.04 , 0.49 ± 0.01 and 0.69 ± 0.02 , respectively). As 4S 4 and STI571 had more inhibition on the K562 cells viability than herbimycin (P< 0.01 ), and there was no statistically significant difference between that of As 4S 4 and STI571. The effects of 1.0, 5.0 and 10.0 μmol/L As 4S 4 on K562 cells were time dependent. When the concentration was lower than 1.0 μmol/L, As 4S 4 had little effect on K562 cells. After being cultured with 2.0 μmol/L As 4S 4 for 24 to 48 hrs, typical morphological changes of apoptosis appeared in the K562 cells. After being cultured with 5.0 μmol/L As 4S 4, STI571 and herbimycin for 72 hrs, the apoptosis rate of K562 cells were 68.8%, 56.7% and 35.5%, respectively. When the concentrations of As 4S 4 changed from 2.0 to 3.0 μmol/L, the apoptosis rate increased from 25.7% to 45.3%. There was no significant difference between the apoptosis rate of K562 cells induced by 5.0 and 10.0 μmol/L As 4S 4. Conclusions As 4S 4 with the concentration of 2.0 μmol/L could inhibit the growth of K562 cells efficiently through inducing apoptosis.

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Objective Substance P (SP) is an important transmitter of non cholinergic excitatory nerves in the lung and can cause neurogenic inflammation in the airway of asthma. It is not know whether inhalation of glucocorticoids (GCs) can decrease the SP contents in lungs. This paper aims at studying the effect and molecule mechanism of GCs on the SP level. Methods Thirty guinea pigs were used to establish asthma models and were randomly assigned into a GCs treated asthma group and an Asthma group without treatment (n=15 each). The GCs treated asthma group received the beclomethasone dipropionate (BDP) aerosolized solution on the day before asthma inducement, on the day of inducement and 24 hrs after inducement. The SP contents in the plasma, brorchoalveolar lavage fluid (BALF) and lung tissues were detected by radioimmunoassay and the SP mRNA expression in the lung tissues was assayed by RT PCR technique 24 hrs after asthma inducement. Fifteen normal guinea pigs, which inhaled normal saline, were used as the Normal control group. Results The SP contents in the plasma, BALF and lung tissues (122±46 pg/ml, 90±39 pg/ml, 78±15 pg/g) in the Asthma group without treatment were significantly higher than those of the Normal control group (84±33 pg/ml, 32±21) pg/ml, and 42±12 pg/g respectively) and the GCs treated asthma group (50±13 pg/ml, 47±20 pg/ml and 40±13 pg/g respectively) (all P< 0.01 ). The plasma SP content in the GCs treated asthma group was lower than that of the Normal control group (P< 0.05 ), while the BALF and lung tissue SP contents did not differ from the Normal control group. The SP mRNA expression in the lung tissues in the Asthma group without treatment ( 1.0 ± 0.02 ) was significantly higher than that of the Normal Control group ( 0.2 ± 0.05 ) and the GCs treated asthma group ( 0.3 ± 0.06 )( both P< 0.01 ), and no significant difference was found between the GCs treated asthma group and the Normal control group. Conclusions Glucocorticoid can significantly down regulate SP mRNA expression and decrease the SP contents in plsma, BALF and lung tissues in asthmatic guinea pigs.

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Objective Oxygen therapy is a common means in the treatment of lung disease in premature infants. However, long time inhalation of high concentration of oxygen can induce lung injury, even chronic lung disease (CLD). This paper aims at studying the dynamic changes of lung ultrastructure and oxidative stress reaction in premature rats with hyperoxia induced CLD and in exploring the role of lung oxidative stress reaction in pathogenesis of CLD. Methods Eighty premature rats were randomly assigned into a Hyperoxia group and a Control group (n=40 each). The Hyperoxia group was given a high concentration of oxygen (FiO 2>0.90) and developed CLD. The Control group received oxygen with the FiO 2 of 0.21. The superoxide dismutase (SOD) activity and malondialdehyde (MDA) concentration of the lung were assayed with a spectrophotometer and the lung ultrastructure in the two groups was observed under a transmission electronmicroscope. Results The structures of mitochondria and lamellar bodies in alveolar epithelial cell Ⅱ (AEC Ⅱ) were partly damaged on the 3rd day of hyperoxia induction. The abnormal nuclei developed in addition to the damaged mitochondria and lamellar bodies in AEC Ⅱ after the 7th day. Compared with the Control group, the MDA level in the Hyperoxia group was increased significantly on the 3rd day ( 55.9 ± 5.5 nmol/mg vs 22.5 ± 4.4 mmol/mg) (P< 0.01 ), and it reached a peak on the 7th day ( 94.3 ± 12.4 nmol/mg). After 7 days the MDA level in the Hyperoxia group decreased gradually but still remained at a higher level on the 21st day than that of the Control group ( 48.0 ± 7.5 nmol/g vs 23.6 ± 5.7 nmol/g) (P< 0.01 ). The SOD activity in the Hyperoxia group did not differ from the Control group. Conclusions AEC Ⅱ injury is early manifestation of hyperoxia induced CLD. The lung oxidative stress reaction is closely related to the AEC Ⅱ injury.

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To study the potential therapeutic effect of Salamarum on neonatal hypoxic ischemic encephalopathy (HIE). Methods Sixty two neonates with moderate HIE were randomly assigned into two groups: a Control group (n=30) receiving conventional treatment and a Treatment group (n=32) receiving salamarum treatment, using loading dosage of 200-250 mg/kg and maintenance dosage of 125 mg/kg daily for 72 hrs after admission. The disappearance time of the nervous system symptoms, the neonatal behavioral neurological assessment (NBNA) scores and the development quotient (DQ) of Gesell Development Schedules for 6-12 months old infants were recorded. Results The time to cessation of gaze and the recovery time of consciousness in the Treatment group was not significantly different from that of the Control group. The time to cessation of convulsions and the recovery time of primitive reflexes and muscular tension were significantly shorter in the Treatment group. More neonates in the Treatment group had normal scores of NBNA than the Control group at 7 and at 12-14 days of life (P< 0.05 ). Moreover, the DQ values in the Treatment group were higher than the Control group at 6-12 months of life ( 91 ± 15 vs 83 ± 15 ) (P< 0.05 ). Conclusions Early salamarum administration can shorten the disappearance time of nervous system symptoms and reduce sequelae in neonates with HIE.

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Objective The immunophenotype, which can identify origins and differentiation phases of cells, is an important biological character of childhood acute leukemia (AL). This paper aims at studying the characteristics and distribution of childhood AL. Methods The cell surface or cytoplasma antigens were detected by flow cytometry with a standard direct or indirect immunofluorescence assay in 160 children with AL. Results Among the 160 cases of AL, 112 were acute lymphoblastic leukemia (ALL) and 48 were acute myelogenous leukemia (AML). Among the 112 cases of ALL, 12 were T lineage ALL (T ALL) ( 10.7% ) and 93 were B lineage ALL (B ALL) (apart from mature B ALL) ( 83.0% ); the myeloid antigen expression was noted in 13.8% of the cases with the most common expression of positive CD33. Among the 48 cases of AML, the lymphoid antigene expression was positive in 40%, with the highest frequency of CD7. The frequency of the lymphoid antigen expression in AML cases was higher than that of the myeloid antigen expression in ALL cases (P< 0.01 ). The frequency of CD14 expression in childhood M4 and M5 AML was 77.8% . The positive CD 34 of the B ALL cases was found in 58.5% of the cases, and that of the AML cases in 55.3% of the cases, in which those of M1 and M2 AML cases was 76.5% , higher than those of the cases with other types of AML ( 43.3% ) (P< 0.05 ). The HLA DR expression was 11.1% in M 3 AML cases, lower than that in the other types of AML cases ( 76.3% ) (P< 0.01 ). Among the B ALL cases, the CD34 expression was un related to the French American British classification, initial WBC counts and extramedullary infiltration. The incidence of infantile leukemia in cases with negative CD34 ( 17.6% ) was higher that in cases with positive CD34 ( 4.2% ) (P< 0.05 ). Conclusions The detection of HLA DR expression may be helpful in the diagnosis of M3 AML.

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The detection of the levels of urinary catecholamines metabolites, vanillylmandelic acid (VMA) and homovanillic acid (HVA), is an important approach to the early diagnosis of neuroblastoma (NB). The previous measurment of chromatometry, however, was usually affected by many factors as result of an inaccurate findings. This paper aims to study the significance of measuring urinary VMA and HVA levels in children by high performance liquid chromatography (HPLC) in the early diagnosis of NB. Methods Fifty healthy children (Normal control group), 27 children with NB (NB group) and 15 children with ALL (ALL group) were enrolled in this study. The levels of VMA and HVA in random urine were measured by HPLC with the flow phase of pH of 4.3 , 2% methanol and IPRB8 of 3.0 mmol/L and the creatin (Cr) level in random urine was measured by chromatometry. VMA and HVA were expressed as VMA/Cr and HVA/Cr. Results The values of VMA/Cr and HVA/Cr in random urine of the NB group ( 51.60 ± 4.53 and 58.00 ± 3.75 μmol/μmol Cr, respectively) were significantly higher than those of the Normal control group ( 8.42 ± 3.61 and 10.12 ± 3.88 μmol/μmol Cr, respectively) and the ALL group ( 8.78 ± 3.50 and 11.50 ± 2.68 μmol/μmol Cr, respectively) (P< 0.05 ). The values of VMA/Cr and HVA/Cr being higher than 17.5 μmol/μmol Cr and 19.8 μmol/μmol Cr respectively, the levels of those of the Normal control group plus 2 standard deviations, were considered as abnormal. As a result, the diagnosis rate of abnormal VMA or HVA for NB was 96.3% . Conclusions The detection of VMA and HVA levels in random urine by HPLC can provide reliable experimental evidence for early diagnosis and differential diagnosis of NB.

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Objective To study the teratogenicity of valproic acid (vpa) in Wistar rats. Methods Twenty four pregnant Wistar rats were randomly assigned into 5 groups: 3 VPA of different dosages groups (400 mg/kg, 450 mg/kg and 500 mg/kg groups), the normal saline (NS) control group and the Blank control group. On the 9th day of gestation, 3 VPA groups were subcutaneously injected different dosages of vpa at 9am and 4pm. The NS control group was administrated 1 ml of NS instead and the Blank control group had no intervention. On the 20th day of gestation, the embryos were taken out and the specimens from spines were double stained with Alcian blue GX and Alizarin red S. The distance of the two cartilaginous ends of the vertebra arch was measured with a stereomicroscope and compared with the normal limit. Results In the Blank control group, the distance of the two vertebral cartilaginous ends was less than 166.4 μm (the superior limit of 112.0±1.96×27.7 μm) from the 9th thoracic to the 3rd sacral vertebra; less than 185.5 μm (the superior limit of 127.7±1.96×29.5 μm) in the 4th sacral vertebra; and less than 198.7 μm (the superior limit of (142.1±1.96×28.9 μm) in the 5th sacral vertebra. There was no malformation in the NS control group. The incidence of spina bifida occulta was 80%, 93% and 100% in the VPA 400 mg/kg, 450 mg/kg and 500 mg/kg groups respectively. The craniofacial, skull, spine and tail malformations occurred in a fetus in the vpa 500 mg/kg group. Conclusions Vpa could induce spina bifida occulta in foetal rats.

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This is the first reported case of CCHS in China. The infant, of gestation age 36 weeks, had dyspnea immediately after birth. Through the course of the disease, the infant manifested hypercapnia and hypoxemia induced by hypoventilation. These manifestations were improved by mechanical ventilation. However, abnormal blood gas findings were noted after weaning of ventilatory support or by altering ventilator parameters. On day 6 of his life, the infant's autonomous breathing druing wakefulness improved, with an increase in respiratory rate to 36 times per minute, although hypoventilation still existed. However, during sleep, there was a decrease in the respiratory rate to 15 times per minute, as well as arrhythmia and sternal retractions. The infant survived for 14 days on ventilation support. These symptoms are consistent with previously overseas reported cases of CCHS.

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目的　制定定量估测新生儿缺氧缺血性脑病 (HIE)预后的指标以较精确估测HIE的预后 ,并为早期干预治疗提供依据。方法　分析 1 993年 1月至 1 997年 6月由HIE致残及致死患儿的临床资料 ,并与非残儿进行对比。按Fisher判别原理优选生后 1分钟阿氏评分 (X1 )、窒息复苏时间 (X2 )、惊厥持续天数 (X3 )、急性期CT改变 (X4)及治疗 3d后原始反射情况 (X5)作为判别指标 ,建立判别方程式 ,求得判别值ZO =77。以Z >77作为预后不良的标准 ,回代全部病人 ,计算估测的敏感性、特异性及准确性。 1 997年 8月至 2 0 0 0年 4月对 4 5例患儿以Z值进行预测 ,随访验证预测结果。结果　以Z >77作为预后不良的标准 ,回代全部病人 ,其估测的敏感性、特异性及准确性分别为 84 .37%、96 .0 5 %和 92 .5 9% ,经对 4 5例患儿的临床验证发现其阳性预测准确度为 90 .91 % ,阴性预测准确度为 94 .2 0 %。结论　以Z >77作为判断新生儿缺氧缺血性脑病预后不良的标准具有较高的应用价值

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目的　先天性甲状腺功能减低症 (CH)是一种严重危害儿童健康、但早期诊治可以有效防治的疾病 ,1 996年以来 ,该院开展了新生儿CH筛查工作 ,现总结 7年来筛查结果 ,以了解湖南省新生儿CH的发生情况。方法　对 91 375例新生儿在生后 72h后或母乳喂养 2 4h后采足跟血 ,采用时间分辨荧光免疫法测定足跟血TSH浓度进行初筛 ,并用化学发光法检测初筛阳性患儿血清中TSH、FT3、FT4以确诊。确诊为CH者立即给予口服甲状腺素片治疗并定期随访。结果　TSH初筛 2 80例阳性 ,初筛阳性率为 1 /32 9,确诊先天性甲低 5 6例 ,发病率为1 /1 6 32。经治疗的患儿智能体格发育均正常。结论　新生儿筛查是早期发现、及时诊断和治疗CH的重要措施 ,能有效地改变患儿的预后

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目的　总结对不同年龄段的学龄前期儿童先天性髋脱位采用综合治疗的临床方法与经验。方法1岁以内采用手法复位并蛙式石膏托外固定 ,1～ 3岁在全麻下行内收肌切断术 ,复位加髋人字石膏固定 ;4～ 6岁采用手术治疗。结果　该组患儿依据标准评定 :优良率为 93.3% ,有 3个髋关节发生股骨头坏死。结论　早期发现、早期治疗是取得满意疗效的重要措施之一 ;合理的综合治疗方案是患儿康复的基础。

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