OBJECTIVE: In order to develop a primary exploration of the relationship between glutathione S transferase (GST) polymorphism and tumor susceptibility or therapeutic correlation in Han nationality, the genetic polymorphism of GST in healthy Han nationality of Shanghai was investigated and the candidate's single nucleotide polymorphism (SNP) was screened. METHODS: Automatic sequencing with labelled fluorescence was used to screen the SNPs of GSTT1 and GSTM1 in 20 healthy Han people. RESULTS: Point mutation was found at 86 057 site between exon 4 and exon 3 of GSTT1 in 4 cases, with guanine G substituted by adenine A. It may be a new candidate's SNP of GSTT1 after compared with genebank's SNP database. G&A heterozygotes could be found at 793 site and 921 site of exon 5 in all cases. Many candidate's SNPs were discovered in all 8 exons of GSTM1 and most of them were heterozygotes. 40% of the examinees showed adenine A deletion in exon 2. All examinees were A&G heterozygotes at 1 383 site and C&G heterozygotes at 1 385 site. At 101 site, 60% of the people examined showed A&T heterozygotes and 40% of the people had adenine A homozygotes. We found deletion of many base pairs or short fragments after 190 bp of exon 2. CONCLUSIONS: The genetic polymorphism of GST varies greatly in healthy Han people in Shanghai. It remains to be discussed further whether the polymorphisms only occurred in Han nationals and correlated with tumor susceptibility and whether these possible candidate's SNPs could alter amino acid code resulting in variation of the primary structure of protein and alteration of GST acitivity.

OBJECTIVE: To investigate the expression and significance of Bcl 2 and MDR 1 genes in children with acute leukemia. Methods The expressions of Bcl 2 and MDR 1 genes in 36 cases of acute leukemia and 10 cases of idiopathic thrombocytopenic purpura in the control group were examined by RT PCR. RESULTS: The expressions of Bcl 2 in the incipient group and relapsed group were higher than that of the control group (P< 0.05 ). While in the incipient group and complete remission group, they were lower than that of the relapsed group (P< 0.01 ). The expression of MDR 1 gene in the relapsed group was higher than those of control group, incipient group and complete remission group (P< 0.01 or 0.05 ). There was no difference of MDR 1 expression between the incipient group and the control group or the complete remission group and the control group (P> 0.05 ). There was no close relationship between the levels of Bcl 2 or MDR 1 and clinical features such as gender, age, initial WBC count, the percent of carcinocytes, hepatomegaly, splenomegaly and lymphadenhypertrophy (P> 0.05 ). The relationship between Bcl 2 and MDR 1 genes was not significant (r s= 0.308 , P> 0.05 ). CONCLUSIONS: Bcl 2 and MDR 1 genes were associated with drug resistance by different mechanisms.

OBJECTIVE: To explore the changes and significance of tumor necrosis factor alpha (TNF α) in the serum and cerebrospinal fluid (CSF) of children with acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML). Methods TNF α was measured by radioimmunoassay in 31 cases of childhood acute leukemia of various phases. RESULTS: Serum TNF α levels pre therapy in ALL and AML [( 24.35 ± 4.84 ) pmol/L, ( 28.65 ± 5.12 ) pmol/L] were significantly higher than that of control [( 11.28 ± 1.69 ) pmol/L], P< 0.01 . Right after complete remission (CR), TNF α decreased [( 16.42 ± 2.57 ) pmol/L, ( 14.57 ± 3.64 ) pmol/L] but was higher than that of control (P< 0.05 ). 6, 12, 24, 36 months after CR, serum TNF α levels in ALL and AML returned to normal. Serum TNF α increased again and was higher than that of control (P< 0.01 ) when relapsed. CSF TNF α pre therapy in ALL and AML [( 12.35 ± 1.74 ) pmol/L, ( 14.56 ± 1.92 ) pmol/L] were also significantly higher than that of control [( 7.54 ± 0.96 ) pmol/L] (P< 0.01 ). During CR and continuous CR, CSF TNF α in ALL and AML patients remained at the level of control (P> 0.05 ). CSF TNF α level in children with central nervous system leukemia (CNSL) was higher than those without CNSL [( 25.62 ± 7.14 pmol/L vs ( 12.15 ± 0.89 ) pmol/L], P< 0.01 . There was a positive correlation between white blood cell count and TNF α level in the CSF (r= 0.942 , P< 0.05 ). CSF TNF α level decreased gradually after intrathecal therapy, but it decreased more slowly than the white blood cells of CSF. CONCLUSIONS: TNF α concentration in the serum and CSF may be of great value in reflecting leukemic cell burden, early diagnosis of CNSL and monitoring intrathecal chemotherapy.

OBJECTIVE: To investigate the effects of tumor necrosis factor α (TNF α) and inhibitor protein of nuclear factor κB (IκBα) on pulmonary hemorrhage (PH) of neonatal rats induced by lipopolysaccharide (LPS). METHODS: Seven to ten days old Wistar rats were randomly separated into 2 groups: LPS group, the rats of which were injected with LPS intraperitoneally at the dosage of 5 mg/kg, and they were sacrificed 30 min, 1, 2, 4, 8, 16 and 24 hs after injection of LPS, respectively; normal saline (NS) group, where equal amount of NS was injected intraperitoneally. The number of each time point was 6. The lungs of the rats were examined by eyes and microscope. The protein and mRNA of TNF α in the lung tissues were detected by the method of ELISA and reverse transcription polymerase chain reaction. The expression of IκBα protein was measured by Western Blot. RESULTS: One hour after administration of LPS, patch like bleeding was observed in more than two lobes of the lung. From the 1st h after injection of LPS, TNF α protein increased and was significantly higher than that of NS control group (P< 0.01 ), it peaked at the 2nd h. TNF α mRNA increased 30 min after injection of LPS and was significantly higher than NS control (P< 0.01 ) and the peak was reached 2 hs after injection of LPS. The expression of IκBα quickly weakened from the 1st h after injection of LPS (P< 0.05 ), it was the lowest at the 8th h. CONCLUSIONS: LPS may lead to lung hemorrhage in the neonatal rats, which may be associated with the decrease of IκBα protein in the lung tissue and activation of NF κB, up regulation of the transcription of TNF α gene.

OBJECTIVE: To inquire into the effects of angiotensin Ⅱ type 1 receptor antagonist (AT1RA) valsartan on apoptosis and the expression of apoptosis correlated protein Fas and FasL in the kidneys of rats with nephrotic glomerulosclerosis induced by adriamycin. METHODS: Thirty six SD rats were randomly divided into model group, treatment group and control group. The model was established by uninephrectomy and injection of adriamycin. In the treatment group, valsartan (20 mg/kg) was delivered daily by gavage for 12 weeks. The same amount of normal saline was delivered by gavage in the control and model groups. Apoptosis was examined by means of terminal deoxynucleotidyl transferase mediated d UTP nick end labeling (TUNEL). Glomerular apoptotic index (GAI) and renal tubule apoptotic index (TAI) were calculated. Immunohistochemistry was utilized to detect the expression of Fas and FasL. The morphological changes were observed under optic microscope and the glomerulosclerosis index (GSI) was determined. RESULTS: Compared with the control group, the GSI, GAI, TAI and the expression of Fas and FasL in model and treatment groups were stronger (P< 0.01 ). Compared with model group, they decreased in treatment group (P< 0.01 ). CONCLUSIONS: Valsartan may suppress the excessive apoptosis of kidney cells by lowering the expression of Fas and FasL so as to postpone the process of glomerulosclerosis.

OBJECTIVE: To understand if there are positive nestin cells in the embryonic human brain. METHODS: Fifteen normal embryoes with gestational ages of 16-32 weeks due to termination of pregnancy were enrolled in this study. The cells from hippocampus, striatum and ependyma in the embryonic human brain were studied using HE staining and immunocytochemistry. RESULTS: Different amounts of positive nestin cells were found in the hippocampus, striatum and under ependyma of the embryonic human brain with gestational age of 16 w, 17 w, 20 +3 w, 22 w, 23 w, 24 +5 w, 25 w and 27 w respectively. The positive nestin cells were displayed two different types in morphology, more in small round form than in big oval shap one. CONCLUSIONS: There are positive nestin cells, with two different forms, in the embryonic human brain.

OBJECTIVE: To study plasma neuropeptide Y (NPY) and β Endorphin (β EP) levels in neonates with asphyxia and to explore the relationship between the two indexes and the degrees of asphyxia, as well the post asphyxia brain damage.METHODS: The plasma levels of NPY and β EP were measured in 37 asphyxia and 12 normal neonates by radio immunoassay. The cerebral CT scan was taken and the CT values were measured. RESULTS: The plasma levels of NPY and β EP in the severe asphyxia neonates were much higher than those of the normal ones [( 1.85 ± 1.10 ) μg/L vs ( 0.04 ± 0.03 ) μg/L and ( 2.03 ± 1.45 ) μg/L vs ( 0.06 ± 0.04 ) μg/L respectively; P< 0.01 ]; also higher than those of the mild ones [( 0.47 ± 0.38 ) and ( 0.34 ± 0.33 ) μg/L respectively] (P< 0.01 ). There were significant differences in the levels of NPY and β EP between the mild asphyxia neonates and the normal neonates (P< 0.01 ). The CT values of the mild and severe asphyxia neonates [( 15.60 ± 2.20 ) and ( 13.08 ± 2.18 ) Hu] were obviously lower than that of the normal neonates [( 20.16 ± 2.66 ) Hu] (P< 0.01 ). The CT value of the severe asphyxia neonates group was obviously lower than that of the mild ones (P< 0.01 ). A positive correlation was found between NPY and β EP levels in the severe asphyxia neonates (r= 0.4220 ,P< 0.01 ). CONCLUSIONS: The plasma levels of NPY and β EP and cerebral CT value were closely related with the degrees of asphyxia. The more severe the asphyxia , the higher the plasma levels of NPY and β EP, and the lower the CT value. The plasma NPY and β EP may be used as the markers in evaluating the degrees of neonatal asphyxia and post asphyxia brain damage.

OBJECTIVE: To study the effect of nonprotein bound iron (NPBI) on neonatal postasphyxial reperfusion injury. METHODS: Plasma concentrations of NPBI from 20 asphyxiated newborns were measured serially with the matrix effect free bleomycin assay during 0 to 6 hours, 6 to 12 hours and 12 to 72 hours after birth. They were then compared with those from 20 healthy newborns within 6 hours after birth (controls). RESULTS: The positive NPBI rate was significantly higher in asphyxiated newborns than that in the controls (80% vs 20%). NPBI was significantly elevated in asphyxiated newborns during 0 to 6 hours [( 4.14 ± 2.41 ) μmol/L] and 6 to 12 hours [( 2.26 ± 2.21 ) μmol/L] compared with that in the controls [( 0.28 ± 0.79 ) μmol/L] (P< 0.05 ). The highest level of NPBI was noted during 0 to 6 hours after birth. CONCLUSIONS: NPBI may play an important role in neonatal postasphyxial reperfusion injury.

OBJECTIVE: To study the infancy symptoms of premature birth children with cerebral palsy so as to provide a criterion for early screening and diagnosis of cerebral palsy.METHODS: The incidence of cerebral palsy and its infancy symptoms were investigated in 4 167 pre term birth children ranging from 1-6 years old from 7 counties/cities of Jiangsu Province. Pediatricians and health care doctors finished the investigation.RESULTS: The average incidence of cerebral palsy in pre term birth children was 32.9‰ . The incidence of cerebral palsy in pre term birth children with the gestational age of less than 32 weeks was 3.6 times as much as that in ones with the gestational age of 34-37 weeks. The positive rate of infancy symptoms of cerebral palsy was 30%-62%, with the specificity over 99.4% and veracity over 97.2% . The positive expected value was over 80%, and negative one was over 97.5% . CONCLUSIONS: The infancy symptoms of cerebral palsy can be used for early screening and diagnosis for cerebral palsy in premature birth children.

OBJECTIVE: To study the relationship between the change of interleukin 5 (IL 5) level in induced sputum and degrees of asthma attack in children with acute asthma, and to study the role of IL 5 in pathogenesis of asthma. METHODS: Sixty five cases of acute asthma were divided into 3 groups according to the degrees of asthma attack: mild, moderate and severe groups. Thirty four healthy children were served as the control group. Ultrasonically nebulized hypertonic saline was used to induce sputum. The IL 5 level in induced sputum was determined using ELISA. The forced expiratory ratio (FEV 1) and the eosinephils (EOS) counting in the sputum were measured. RESULTS: The EOS counting and IL 5 level in the induced sputum of children with acute asthma attack increased compared with those in the control group, whereas the FEV 1 decreased in the former (P<0.01 ). As asthma state became worse, EOS counting in induced sputum increased, but no significant difference was noted among different degrees of athma attack. The IL 5 level in the induced sputum was obviously different among the mild, moderate and severe cases [( 8.8 ± 4.9 ), ( 82.7 ± 173.5 ) and ( 225.1 ± 228.9 ) pg/ml, respectively] (P< 0.05 ). The FEV 1 in the severe group was significantly lower than those in the mild and moderate groups (P< 0.05 ). The IL 5 level in induced sputum was significantly positively correlated with EOS counting (r= 0.482 ,P< 0.05 ) and significantly negatively correlated with FEV 1 (r= -0.647 ,P< 0.01 ). CONCLUSIONS: Compared with EOS counting, the IL 5 level in induced sputum can more accurately reflect the degrees of asthma attack, indicating that it may be used as a sensitive index for evaluating the asthmatic state of children and the therepeutic effectiveness of drugs.

OBJECTIVE: The incidence of tuberculosis (TB) has begun to rise again in the world in recent years. Reseachers have focused mainly on treatment for TB but have seldom studied on protective effect of vaccination of bacillus calmette guerin (BCG) on children. The aim of this study was to investigate the protective duration and impact of vaccinating children with BCG. METHODS: The BCG scar rate and positive purified protein derivative (PPD) rate were studied in 9 608 children who were vaccinated in both the urban and rural areas of Xianning City and who ranged in age from 0 to 14 years. RESULTS: The positive PPD rate of the children between 3 months and 1 year was the highest ( 91.5% ); it was the second highest in children ranging from 1 to 3 years ( 81.6% ), the lowest in children ranging from 3-7 years ( 57.1% ) and rose again in children ranging from 7-14 years ( 74.2% ) (P< 0.01 ). The positive PPD rate of the urban children was higher than that of the rural children ( 78.8% vs 62.4% ); the positive BCG scar rate of the urban children was 90.3% , higher than that of the rural children ( 56.1% ) (P< 0.01 ). The inciderce of natural TB infection in urban children was lower than that of the rural ones ( 1.7% vs 3.4% ). The incidence of natural TB infection in the children between 3 months to 3 years old was higher than those of the children raning from 3-7 years and 7-14 years. It was the highest in the children ranging fro 7 to 14 years.CONCLUSIONS: The protective effect of the BCG vaccination on 0-3 year old children is significant, but reduces on children of 4 years of age or older. The positive PPD rate and BCG scar rate of the urban children were higher than those of the rural children, indicating that the quality of vaccination of BCG should be improved in rural areas.

OBJECTIVE: To study the effect and mechanism of early intervention by polysaccharide nucleic acid fraction of BCG (BCG PSN) for prevention of post bronchiolitis wheezing. METHODS: Forty infants with bronchiolitis were randomly assigned into 2 groups: intervention group (n=19) and non intervention group (n=21). Three months after recovering from bronchiolitis, the infants in the intervention group were given BCG PSN injection 1 ml I.M twice a week, 24 times in all. The infants in the non intervention group did not receive any intervention. Twenty healthy children were used as the normal controls. The levels of IFN γ and IL 4 in the supernatant of cultured peripheral blood mononuclear cells, and serum IgE level were measured using ELISA before intervention and one year after intervention. One year following up was done to observe the frequency of the wheezing episodes. RESULTS: The IFN γ level and IFN γ/IL 4 ratio were significantly lower, and the levels of IL 4 and IgE were significantly higher in both intervention group and non intervention group before intervention than those in the normal controls (P< 0.01 ). After one year of BCG PSN intervention, the IFN γ level [( 986.95 ± 363.20 ) ng/L] and IFN γ/IL 4 ratio ( 16.03 ± 7.11 ) were significantly higher than those of before intervention [( 768.43 ± 288.07 ) ng/L and ( 9.11 ± 4.27 ) respectively]; the levels of IL 4 [( 59.73 ± 16.04 ) ng/L] and IgE [( 48.05 ± 19.57 ) kU/L] decreased significantly compared with those of the pre intervention [( 81.66 ± 21.92 ) ng/L and ( 155.35 ± 76.25 ) kU/L, respectively] (P< 0.01 ) and the IFN γ level was not significantly different from the normal controls (P> 0.05 ). In one year of BCG PSN intervention, 3 cases ( 15.8% ) had two or more times wheezing episodes in the intervention group, less than that in the non intervention group [11 cases ( 52.4% )] (P< 0.05 ).CONCLUSIONS: Early intervention with BCG PSN in post bronchiolitis children may be useful in preventing the infants from recurrent wheezing episodes later. The possible explanation is that BCG PSN can increase IFN γ secretion, inhibit IL 4 activity, increase IFN γ/IL 4 ratio and decrease IgE production.

OBJECTIVE: To study the change of cleaved Caspase-3 expression within 24 hs following hypoxia ischemia brain damage (HIBD) in neonatal rats. METHODS: Seven day old rats were randomly assigned into control group and HIBD group. The Caspase-3 expression was assayed by the immunohistochemical staining and Western Blot analysis at 3, 6, 12 and 24 hs after hypoxia ischemia (HI). RESULTS: The positive staining of Caspase-3 transferred from cytoplasm to the nuclei and the cells were apparently condensed and shrunk within 24 hs after HI. The number of the cleaved Caspase-3 positive cells in the damaged cortex increased at 3 h, decreased at 6 h, increased again at 12 h and peaked at 24 h after HI, whereas the number in the damaged hippocampal CA1 region increased with time and peaked 24 hs after HI. The number of the Caspase-3 positive cell both in the cortex and the hippocampal CA1 region of the HIBD group at 3 h, 6 h, 12 h and 24 h was greater than that of the control group (P< 0.05 or 0.01 ). Caspase-3 expression was also found in the damaged brain tissue 3 hs after HI by Western blotting analysis. The intensity of the expression, decreasing at 6 h but increased again 12 hs after HI. CONCLUSIONS: The character of brain cell apoptosis within 24 hs following HIBD, increase of apoptotic cells twice, may be found with Caspase-3 as the marker for evaluating brain cell apoptosis in HIBD and it may be as reference for using anti apoptotic agents in treatment of HIE.

OBJECTIVE: To explore the role of Caspase-3 protein and DNA fragmentation at late stage after hypoxic ischemic brain damage (HIBD) and the effect of basic fibroblast growth factor (bFGF) on Caspase-3 expression. METHODS: Seven day old Wistar rats were randomly assigned into HIBD group, sham operated group, normal saline treatment group and bFGF treatment group. Caspase-3 expression of the brain was measured by the immunohistochemical method and DNA fragmentation of neurocytes was detected by TUNEL at 2 w, 3 w or 4 w after hypoxia ischemia (HI). RESULTS: ① The number of the TUNEL positive cell in the HIBD group at 2 w and 3 w after HI was greater than that of the sham operated group (P< 0.05 ). ② The Caspase-3 average OD values of the brain ( 0.39 ± 0.04 , 0.38 ± 0.04 ) in the HIBD group at 2 w and 3 w after treatment were significantly higher than those in the sham operated group ( 0.36 ± 0.01 , 0.36 ± 0.02 ), P< 0.05 . ③The Caspase-3 average OD value in the bFGF group at 3 w after HI ( 0.36 ± 0.09 ) was significantly lower than those in the control group and HIBD group ( 0.37 ± 0.04 , 0.38 ± 0.04 ), P< 0.05 . ④ The number of the TUNEL positive cell in the bFGF group at 3 w after treatment ( 4.20 ± 1.30 ) was significantly less than those in the control group and HIBD group ( 9.80 ± 1.92 , 8.00 ± 1.00 ), P< 0.05 . CONCLUSIONS: Caspase-3 may take part in the pathogenesis of HIBD and keep continuous activation, suggesting a prolonged role for apoptosis in neonatal HIBD. bFGF has neuroprotective effects against neonatal HIBD by down regulating Caspase-3 protein expression and preventing DNA from fragmentation.

OBJECTIVE: To study the mechanism of LPS, IL 1β and TNF α induced apoptosis of astrocytes in rats. METHODS: The astrocytes of the neonatal rat were cultured in vitro and randomely assigned into the inflammation group (treated with LPS, IL 1β and TNF α), pre treatment group (L NMMA was administrated half an hour before treatment with LPS, IL 1β and TNF α) and normal control group. The cell viability was assayed by MTT; apoptosis was evaluated by electron microscope and flow cytometry; nitric oxide (NO) content was measured by spectrophotometer; cytochrome C content in cytoplasm was measured by Western Blot and Caspase 3 mRNA expression was detected by hybridization in situ. RESULTS: The cell viability in the pre treatment group was significantly higher than that of the inflammation group ( 0.81 ± 0.29 vs 0.46 ± 0.15 ), P< 0.01 . There was a negative correlation between the cell viability and NO content produced by astrocytes in the inflammation group (r= -0.604 , P< 0.05 ). The apoptosis rate of the pre treatment group was lower than that of the inflammation group [( 15.2 ± 7.9 ) % vs ( 29.7 ± 10.4 )%; P< 0.05 ]. The cytochrome C content in cytoplasm of astrocytes of the pre treatment group (14 784±2 096) decreased compared with that of the inflammation group (31 049±3 784) (P< 0.01 ). The expression of Caspase 3 mRNA in the inflammation group was intensified compared with that of the pre treatment group. CONCLUSIONS: Cytokines may lead to decrease of viability of astrocytes through increasing NO content produced by astrocytes. The increase of NO content can induce astrocyte apoptosis, which is related to the release of the cytochrome C from mitochondria to cell plasma, and activating the expression of Caspase 3 mRNA.