OBJECTIVE: This study was designed to investigate the effect of human cytomegalovirus(HCMV) on the proliferation of colony forming unit granulocyte-macrophage(CFU-GM),CFU-erythroid(CFU-E),burst forming unit-erythroid(BFU-E),CFU-multipotential(CFU-Mix) and CFU-megakaryocytic(CFU-Mk) progenitor cells of cord blood in vitro as well as the possible mechanism. METHODS: Twenty cord blood specimens were collected from the umbilical vein of normal full-term neonates delivered spontaneously.This study consisted of five groups: 3 Infection groups in which 0.1 mL 10~3,10~4 and 10~5 plague forming unit(PFU) HCMV-AD_(169) virus solution was added to the culture system,an Inactivated control group in which the equal volume of inactivated virus solution was added,and a Blank control group(normal progenitor cells culture system without HCMV virus infection).Colony forming unit-assay was applied to detect the effects of HCMV-AD_(169) strain on the colony formation,inhibition rate and colony-maintaining duration of CFU-GM,CFU-E,BFU-E,CFU-Mix and CFU-Mk of cord blood.PCR technique was used to demonstrate the existence of HCMV-DNA in the colony cells of cultured CFU-GM,CFU-E,CFU-Mix and CFU-Mk. RESULTS: HCMV-AD_(169)(10~3 PFU) in low concentration had inhibition effects on colony formation of the CFU-Mix and CFU-MK(P<0.05),whereas 10~5 PFU and 10~4 PFU HCMV-AD_(169) lead to decreased colonies in CFU-GM,CFU-E,BFU-E,CFU-Mix and CFU-MK compared with the Blank control and the Inactivated control groups(P<0.05).The suppression effect of HCMV on the colony formation was dose-dependant.The colony-maintaining duration of the CFU-GM,CFU-E,BFU-E,CFU-Mix and CFU-Mk in the 10~5 PFU and 10~4 PFU HCMV infection groups was significantly shorter than that in the two control groups(P<0.01).The low concentration of HCMV-AD_(169)(10~3 PFU) infection resulted in a shortened colony-maintaining duration of the CFU-Mix and CFU-Mk(P<0.01),but had no effects on the colony-maintaining duration of CFU-GM,CFU-E and BFU-E.PCR amplification demonstrated the existence of HCMV-AD_(169) DNA in the colony cells of the three Infection groups. CONCLUSIONS: HCMV-AD_(169) strain can infect hematopoietic progenitors of cord blood and inhibit the proliferation of hematopoietic progenitors,associated with anemia,neutropenia and thrombocytopenia in HCMV patients.

OBJECTIVE: This study was designed to investigate the pathophysiological role of adrenomedullin(ADM) in congenital heart disease. METHODS: Forty-eight children with congenital heart disease confirmed by cardiac echocardiography and catheterization were studied.The patients were divided into three groups on the basis of hemodynamic indices measured during cardiac catheterization: high pulmonary blood flow with(group 1) or without(group 2) pulmonary hypertension(mean pulmonary arterial pressure >20 mmHg) and a cyanosis group(without high pulmonary blood flow)(group 3).Six children who recovered from Kawasaki disease were used as a Control group.Plasma ADM levels were measured by radioimmunoassay. RESULTS: The plasma ADM levels from the femoral vein were significantly higher than those from femoral artery in patients with congenital heart disease.The patients from group 1 and group 3 had higher plasma ADM levels(1.9±1.8 pmol/L and 2.4±1.3 pmol/L,respectively) than the controls(1.0±1.4 pmol/L;P<0.01).Plasma ADM levels were significantly negatively correlated with mean systemic arterial pressure,oxygen saturation in mixed vein and oxygen saturation in systemic artery(r=-0.401,-0.562,-0.600,respectively;P<0.01) but positively correlated with pulmonary vascular resistance(r=0.406;P<0.01). CONCLUSIONS: Plasma ADM levels are increased in congenital heart disease with high pulmonary blood flow and hypertension or with cyanosis.Plasma ADM levels are related to pulmonary arterial resistance and hypoxemia.Increased ADM levels may play roles in reducing the pulmonary arterial resistance and alleviating hypoxemia in these patients.

OBJECTIVE: Neuroblastoma is a highly malignant tumor.Stage IV neuroblastoma has a very poor long-term outcome by conventional chemotherapy and surgery and better therapies are essential. This study aimed to explore the long-term effect of high dose induction chemotherapy combined with autologous peripheral blood stem cell transplantation and 13-cis retinoid acid treatment on stage IV neuroblastoma in children. METHODS: Twenty-eight children with stage IV neuroblastoma,aged 2.1-11.5 years(mean 3.3±1.9 years),were employed for the study.Primary sites of the tumors included adrenal(n=23), chest(n=3), chest-abdomen(n=1) and sacrum(n=1).Before autologous peripheral blood stem cell transplantation the patients received 6 courses of intensive induction chemotherapy. During chemotherapy the autologous peripheral blood stem cells were harvested and the tumor excision was done. After transplantation the local radiation and 13-cis retinoid acid therapy were administered. RESULTS: After 6 courses of induction chemotherapy 13 patients got complete remission(CR), 11 got partial remission(PR), and 4 had no response. The 24 patients who received CR or PR completed the full therapy.A 3.5±0.7 years follow-up showed that the 4-year event-free survival of the CR and PR patients was 29.2%.The median no-relapse survival time in CR patients was 4.1±0.7 years but 2.8±0.5 years in PR patients(t= 3.9, P<0.01). CONCLUSIONS: High dose chemotherapy combined with autologous peripheral stem cell transplantation and 13 cis-retinoid acid treatment can improve the long-term outcome of patients with stage IV neuroblastoma. The patients in CR before transplantation had better outcomes than those in PR.

OBJECTIVE: To investigate the effects of survivn antisense oligonucleotide(ODN) on cell proliferation and apoptosis of HL-60 cells. METHODS: Synthetic ODN was completely phosphorothioate-modified. Cationic lipid-mediated antisense ODN was transferred into HL-60 cells. The expression of survivin mRNA and protein was detected by RT-PCR and Western Blot.The incorporation of MTT was used as the measurement of HL-60 proliferation. The cell-cycle and apoptosis were analyzed by flow cytometry. RESULTS: HL-60 cells spontaneously expressed survivin mRNA and protein. Both mRNA and protein expression of survivin decreased significantly in the antisense ODN transfected cells in comparison to that in the original cells and cells transfected with sense ODN.Survivin antisense ODN significantly inhibited cell proliferation and induced apoptosis in a dose-dependent manner. The cell-cycle in the antisense ODN-transfected cells stopped at the G_2/M phase. CONCLUSIONS: Antisense ODN targeting at survivin mRNA can inhibit HL-60 cell proliferation and induce G_2/M stop and apoptosis.

OBJECTIVE: Survivin,a unique member of the inhibitor of apoptosis protein(IAP) family, plays an important role in regulating both apoptosis and cell division. Overexpression of survivin is associated with increased risk of recurrence and poor outcome in cancer patients. This study aimed to investigate the expression of survivin and its location as well as the relationship between cellular location and expression of surrivin and the therapeutic efficacy at the cellular level. ETHODS: The expression of survivin protein was deuected by immunohistochemical assay in bone marrow cells from 62 children with acute leukemia and 40 hospitalized children did not have leukemia (Control group), and in a human acute T lymphocytic leukemia cell line(Molt-4 cells) treated in vitro with daunorubicin(DNR). Cell apoptosis was detected using flow cytometry. RESULTS: Survivin protein was expressed in 41.9% of the 62 children with acute leukemia but in only 5.0% of the Control group(χ~2=16.66; P <0.01). The expression rate of survivin was 46.2% in cytoplasm and 53.9% in nucleus in the children with acute leukemia(χ~2=0.3077; P>0.05). However, the remission rate of patients in whom survivin expression was seen in the nucleus was significantly higher than that in patients in whom survivin was expressed in cytoplasm after chemotherapy. The survivin expression in Molt-4 cells decreased remarkably by DNR treatment in a time and dosage-dependent manner. DNR treatment also induced survivin transllocation from cytoplasm to nucleus and cell apoptosis in a time and dosage-dependent manner. CONCLUSIONS: Survivin may play an important role in the development and prognosis of childhood acute leukemia. The different expression pattern of survivin in the cytoplasm and the nucleus may be associated with therapeutic efficacy and prognosis in acute leukemia. DNR may reduce the survivin expression in leukemic cells and induce cell apoptosis.

OBJECTIVE: This study aimed to explore the effects of hyperoxia on the development of fetal lung by investigating the changes of morphological and cell proliferation induced by hyperoxia in cultured fetal lungs as well as the effects of dexamethasone on hyperoxia-exposed lungs. METHODS: Human fetal lung explants at the pseudoglandular stage of development were cultured randomly either in normoxia(21% O_2/5% CO_2) or hyperoxia(95% O_2/5% CO_2) for 72 hrs. Dexamethasone was added into the feeding medium at the concentration of 10~(-6) M. Harvested tissues were stained for pancytokeratin to identify epithelial cells, with Ki-67 as a marker of proliferation. The effects of lung morphometry were analyzed using computer assisted image analysis.The mean airway thickness, the proportion of the surface area occupied by airways, the mean airway surface area and the index of the epithelium proliferation were measured. RESULTS: The lung architectures remained unchanged after 72 hrs normoxia culture, whereas hyperoxia culture resulted in significant dilation of airways and thinning of epithelium, with the surface area of airways of 6 662 μm~2 vs 2 728 μm~2 and the thickness of airways of 7.8 μm vs 8.1 μm(P<0.05). Hyperoxia culture also resulted in an increase in the proportion of the surface area occupied by airways than normoxia culture(35.2% vs 23.4%; P<0.05). The surface area of airways(3 174 μm~2) and the proportion of the surface area occupied by airways(23.9%) decreased significantly in hyperoxia-cultured lungs after dexamethasone administration(P<0.05). The epithelium proliferation index in hyperoxia-cultured lungs(21.8%) was higher than that in normoxia-cultured lungs(5.1%) and dexamethasone-treated hyperoxia-cultured lungs(7.4%)(P<0.05). CONCLUSIONS: The exposure of pseudoglandular lungs to hyperoxia modulates the lung architecture to resemble saccular lungs with higher epithelium proliferation index. Dexamethasone may inhibit the effects induced by hyperoxia.

OBJECTIVE: Interleukin-4 plays a key role in the development of asthma.Overseas studies have shown that Q576R polymorphism in the interleukin-4 receptor(IL-4R) gene is related to asthma as well as increased serum IgE levels. This study was designed to investigate the association of Q576R polymorphism in IL-4R gene with childhood asthma and serum IgE levels. METHODS: The polymorphism of IL-4R Q576R was determined by PCR/RFLP and serum total IgE level was measured using ELISA in 94 children with asthma. Sixty-eight healthy children served as controls. RESULTS: The distribution frequency of heterozygous genotype Q576R(41%) and mutant allele R576(26%) was significantly higher in children with asthma than that of controls(16% each)(P<0.01; P<0.05). The total serum IgE level between patients with genotype Q576R and Q576Q was not significantly different(225.78±51.43 IU/mL vs 163.24±31.32 IU/mL, P>(0.05)). CONCLUSIONS: The mutant R576 allele of IL-4R may be one of the candidate genes for susceptibility to asthma. Allele R576 of IL-4R is related to asthma but is irrelevant to the total serum IgE level in children with asthma.

OBJECTIVE: To identify whether there is an association between an acute attack of childhood bronchial asthma and Chlamydia pneumoniae(CP) infection. METHODS: Serum specific antibodies IgM and IgG to CP were detected by ELISA in 120 asthmatic children with an acute attack and 82 healthy children. RESULTS: Anti-CP IgM was demonstrated in 22 cases(18.3%) and anti-CP IgG was demonstrated in 32 cases(26.7%) out of the 120 asthmatic patients.The incidence of CP infection in asthmatic children was significantly higher than that in healthy controls(3.7%)(P<0.01). Glucocorticoid inhalation treatment alone resulted in a remission of an acute attack of asthma in 15 cases out of the 32 cases with CP infection, but 17 cases required glucocorticoid inhalation treatment together with anti-CP infection treatment(macrolide antibiotics,eg.azithromycin) for remission of asthma attack. CONCLUSIONS: There may be a link between an acute attack of childhood asthma and CP infection.It is thus necessary to detect the CP-specific antibodies in asthmatic children for proper treatment.

OBJECTIVE: This study investigated the pathogen distribution and resistance patterns in childhood urinary tract infection in order to provide references for optimal use of antibiotics in the treatment of this disorder. METHODS: The clinical data of 152 children with community acquired urinary tract infection (urinary culture positive) between December 2001 and December 2004 were studied retrospectively. The bacterial pathogens of urinary tract infection and antimicrobial resistance were analyzed. RESULTS: Gram-negative bacilli was predominant pathogenic bacteria, accounting for 79.0% of the cases, and Escherichia coli (E.coli) was most commonly found (56.2%). Gram-positive cocci accounted for 18.4%, including 15.1% of Enterococcus faecalis. Fungi was rarely seen, accounting for only 2.6%. E.coli had a resistance rate of more than 50% to ampicillin,amoxicillin/clavulate, co-trimoxazole, cefradine, and fosomycin, but a very low resistance rate (<4%) to 3rd generation cefalosporin, nitrofurantoi, azactom and amikacin. Enterococcus faecalis had a low resistance rate (<20%) to ampicillin, vancomycin,penicillin, and nitrofurantoin. CONCLUSIONS: E.coli is the major pathogen in community acquired pediatric urinary tract infection, and Enterococcus has been become another important pathogen. Selection of antibiotics for the treatment of this disorder should base on drug-sensitive test results.

OBJECTIVE: Atrial natriuretic peptide (ANP) is a cardiac hormone with many biological effects. Hypersecretion may lead to hyponatremia. This study examined the umbilical ANP levels in high risk neonates. METHODS: A total of 117 high risk neonates born between June, 2004 and June, 2005 were divided into Simple asphyxia and Normal score groups according to their Apgar's scores. The Simple asphyxia group was subdivided into Mild (n=20) and Severe asphyxia groups (n=17), and the Normal score group was subdivided into Infection (n=25) and Non-infection groups (n=55). Forty normal neonates were used as the Control group. The samples of umbilical cord blood were collected at delivery and the umbilical ANP levels were measured by radioimmunoassay. Meanwhile the sodium levels in the peripheral vein were measured. RESULTS: The mean umbilical ANP levels in high risk neonates were significantly higher than those in the normal neonates. A more significant increase of the umbilical ANP level was observed in premature infants (1.46±(0.39) ng/mL), and neonates with serious infection (1.16±0.35 ng/mL) and with severe asphyxia (2.12±0.46 ng/mL)compared with the normal neonates (0.62±0.33 ng/mL;P<0.01). The serum sodium level was negatively correlated with the umbilical ANP level (r=-0.99,P<0.01). CONCLUSIONS: The umbilical ANP levels increased significantly in the high risk neonates, suggesting high risk neonates are susceptible to hyponatremia.

OBJECTIVE: To explore the genetic susceptibility of children to vitamin D deficiency rickets through studying the association between Vitamin D receptor (VDR) gene polymorphism and vitamin D deficiency rickets. METHODS: One hundred and fifty-nine children (100 boys and 59 girls,aged 0 to 2 years), with new-onset vitamin D deficiency rickets were enrolled. The patients sampled from a community of Jiamusi City, Heilongjiang Province. Seventy-eight healthy age-matched children (46 boys and 32 girls) were used as the controls. VDR gene polymorphism (cleaved by restriction endonuclease FokI) was analyzed by polymerase chase reaction-restriction fragment length polymorphism(PCR-RFLP). The frequencies of the VDR genotype and allele were compared between the two groups. RESULTS: The frequencies of FF, Ff and ff genotypes were 37%, 51% and 12% in the Rickets group, and 18%, 55% and 27% in the Control group. A significant difference was found in the frequency distribution of the VDR genotype between the two groups (χ~2_(0.01(2))=9.210, χ~2=13.3880, P<0.01). In the Rickets group, f allele frequency was lower (37% vs 54%), while the F allele was more common than the Control group (63% vs 46%). CONCLUSIONS: There is an association between the VDR gene Fok I polymorphism and vitamin D deficiency rickets. The individuals with the F allele are more susceptible to vitamin D deficiency rickets.

Neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) is a kind of inborn errors of metabolism,with the main clinic manifestations of jaundice, hepatomegaly, and abnormal liver function indices. As a mitochondrial solute carrier protein,citrin plays important roles in aerobic glycolysis, gluconeogenesis, urea cycle,and protein and nucleotide syntheses. Therefore citrin deficiency causes various and complicated metabolic disturbances, such as hypoglycemia, hyperlactic acidemia, hyperammonemia, hypoproteinemia, hyperlipidemia, and galactosemia. This paper reported a case of NICCD confirmed by mutation analysis of SLC25A13, the gene encoding citrin. The baby (male, 6 months old) was referred to the First Affiliated Hospital with the complaint of jaundice of the skin and sclera, which it had suffered from for nearly 6 months.Physical examination showed obvious jaundice and a palpable liver 5cm below the right subcostal margin.Liver function tests revealed elevated enzymatic activities, like GGT, ALP, AST, and ALT, together with increased levels of TBA,bilirubin (especially conjugated bilirubin), and decreased levels of total protein/albumin and fibrinogen. Blood levels of ammonia, lactate, cholesterol, and triglyceride were also increased,a nd in particular, the serum AFP level reached 319 225.70 μg/L, a extremely elevated value that has rarely been found in practice before. Tandem mass analysis of a dried blood sample revealed increased levels of free fatty acids and tyrosine, methionine, citrulline, and threonine as well. UP-GC-MS analysis of the urine sample showed elevated galactose and galactitol. The baby was thus diagnosed with suspected NICCD based on the findings. It was then treated with oral arginine and mutiple vitamins (including fat-soluble vitamins A, D, E, and K), and was fed with lactose-free and medium-chain fatty acids enriched formula instead of breast feeding.After half a month of treatment, the jaundice disappeared, and the laboratory findings, including liver function indices, blood levels of ammonia, lactate and AFP, were returned to normal level. The baby was followed up for 6 months. It developed well, a nd the abnormal laboratory findings, including MS-MS and UP-GC-MS analysis results, have been corrected, except a slightly elevated lactate level sometimes. SLC25A13 gene mutation analysis for the patient revealed a compound heterozygote of mutation 851del4 and 1638ins23 and therefore NICCD was definitely diagnosed.

OBJECTIVE: This study was designed to investigate the effects of different oxygen inhalation modes on retinal vessels development in neonatal mice in order to provide experimental data for proper oxygen therapy for premature infants. METHODS: A total of 144 postnatal day (P) 7 C57BL/6J mice were randomly assigned into 6 groups according to different oxygen inhalation modes (n=24). Experimental group 1 was exposed to 30%, 40%, 50%, 60% and 75% oxygen in turn for one day respectively, followed by room air exposure for 5 days. Experimental group 2 was exposed to 75%, 60%, 50%, 40% and 30% oxygen in turn for one day respectively, followed by room air exposure for 5 days. Experimental group 3 was exposed to 75% oxygen for 5 days, followed by room air exposure for 5 days. Experimental group 4 was exposed to 75% oxygen for 5 days, 50% oxygen for 2 days and 30% oxygen for 2 days, then room air exposure for 6 days. The supplemental 75% oxygen and room air recovering was performed alternately for the mice in Experimental group 5 for 3 times and then room air exposure for 5 days. The Control group was exposed to room air for consecutive 10 days. The retinal vascular development and proliferation were evaluated by the retinal flat-mounts 4 (ADPase stained retina) and cross-section. RESULTS: The peripheral vascular pattern was clear, and a few avascular areas were seen in the Control group at P12. At P14 the avascular area disappeared. At P17, the entire vascular pattern became completely normal. In the Experimental groups 1, 3 and 5, the central vessels became tortuous and constricted and the central avascular area increased at P12. At P14, neovascularization was seen, peaking at P17 in the Experimental groups 1, 3 and 5. In the Experimental group 4, the central avascular area increased and neovascularization was seen at P14, but the central avascular area was reduced and abnormal neovascularization disappeared, with slight constriction of the deep vessels, at P17. Five days later the vascular pattern became almost normal in the Experimental group 4. The retinal vascular form of the Experimental group 2 was similar to that of the Control group. The average number of neovascular nuclei extending into the vitreous per cross-section in the Experimental groups 1, 2, 3, 4, and 5 and the Control group was 49.50±1.36, 5.17±0.67, 47.68±4.70, 5.74±2.37, 29.15±2.48, and 1.22±0.20 respectively. There were significant differences between the Experimental groups 1, 3, 5 and the Control group (P<0.05). CONCLUSIONS: The effects of different oxygen inhalation modes on the retinal vessels development in neonatal mice were different. The obvious fluctuation of inhaled oxygen concentration and abrupt stop of supplemental oxygen after high levels of supplemental oxygen may severely affect the development of retina vascular, leading to the pathologic changes similar to retinopathy of prematurity.

OBJECTIVE: The expressions of caspase-1 and cytokines activated by caspase-1 are associated with the pathophysiology of many diseases for its proinflammatory and proapototic peculiarity. However its relationship to brain injury of developing rats following recurrent seizures has not yet been identified. This study aimed to investigate the role of caspase1 and cytokines activated by caspase-1 in brain injury of developing rats following recurrent seizures. METHODS: A total of 96 postnatal 20 day Sprague-Dawley rats were randomly assigned into Control and Seizure groups.Seizures were induced in the Seizure group by flurothyl inhalation daily for six days. Brain tissues were sampled at 6 hrs, and at 1, 3, and 7 days after last seizure. The expressions of caspase-1, interleukin (IL) -18 and IL-1β mRNA in the cerebral cortex were detected by RT-PCR.The water content of the brain and the pathological changes of cortex nerve cells were observed. Brain injury was evaluated using a semiquantitative neuropathological scoring system. RESULTS: The levels of caspase-1and IL-18 mRNA in the cerebral cortex of the Seizure group were obviously higher than those in the Control group at 6 hrs, and at 1, 3, and 7 days after seizure (P<0.05 or P<0.01). The expression of IL-1β mRNA in the Seizure group exhibited a biphasic pattern: increased significantly at 6 hrs, and at 1 and 7 days post-seizure (P<0.01), but was not significantly different from the Control group at 3 days post-seizure. Edema, degeneration and necrosis of nerve cells in cerebral cortex, accompanying by inflammatory cell infiltration and apoptosis of nerve cells, were observed under a light microscope in the Seizure group after recurrent seizures. The water content of the brain in the Seizure group increased significantly compared with that in the Control group at 6 hrs, and at 1 and 3 days after recurrent seizures (P<0.01). The Seizure group had significantly higher neuropathological scores than the Control group at each time point (P<0.01). CONCLUSIONS: Caspase-1 and cytokines activated by caspase-1 play an important role in the developing brain injury after recurrent seizures.

OBJECTIVE: p53-induced apoptosis is crucial in the development of hypoxic-ischemia (HI) brain damage and neurodegenerative disorders. Some experimental research has shown that a synthetic inhibitor of p53 can protect neurons against apoptosis. This study aimed to explore the expression of p53 in neonatal mice following HI brain damage and the effect of p53 inhibitor (pifithrin-α, PFT-α) on brain damage. METHODS: HI was induced in 9-day-old mice pups by ligation of left carotid artery and 10% oxygen exposure for 55 minutes.The pups were sacrificed and the brains were taken out at 3, 8, 24, and 72 hrs post-HI. The brains were sectioned and stained with antibody against p53 and microtubule-associated protein 2 (MAP-2). PFT-α was injected intraperitoneally: in experiment 1 immediately after HI with different dosages (1, 2 and 8 mg/kg); in experiment 2 2 mg/kg at different HI times (1 hr before HI, and immediately and 1 hr after HI). Control animals without HI received injections of 0.5% dimethyl sulfoxide. Brain damage was evaluated by gross morphology scoring at 72 hrs after HI. RESULTS: The number of p53 positive cells in the cortex, hippocampus and striatum of the ipsilateral hemisphere increased significantly and peaked at 3-8 hrs post-HI when compared with those of contralateral hemisphere as well as normal controls. The positive cells distributed mainly in the MAP-2 negative area.Both different dosages and different injection time PFT-α treatment did not reduce the extent of brain damage. CONCLUSIONS: The immunoactivity of p53 increased significantly as early as 3 hrs post-HI. The distribution area of p53 expression was consistent with that of brain damage. The p53 inhibitor PFT-α has no protective effects against HI brain damage in neonatal mice.

OBJECTIVE: Febrile seizure(FS) is one of the most common seizure types in children.Our previous studies have demonstrated that both γ-aminobutyric acid B receptor (GABA_BR) and hydrogen sulfide (H_2S) are involved in the pathogenesis of FS.This study was designed to explore the effect of GABA_BR on H_2S / cystathionine-β-synthase (CBS) system in recurrent FS. METHODS: Sixty-four Sprague-Dawley rats aged 21 days were randomly assigned into four groups: Control (37℃ water bath exposure), FS, FS+baclofen (GABA_BR excitomotor), and FS+phaclofen (GABA_BR inhibitor) groups (n=16 each). FS was induced by warm water bath exposure (45.2℃), once every 2 days, 10 times in total. The plasma level of H_2S was detected by the spectrophotometer. The expression of CBS mRNA was examined by in situ hybridization. The expressions of CBS protein was observed by immunohistochemistry. RESULTS: The plasma level of H_2S increased in the FS + baclofen group (427.45±15.91 μmol/L) but decreased in the FS+phaclofen group (189.72±21.53 μmol/L) compared with that in the FS group (362.14±19.71 μmol/L). The expressions of CBS mRNA and protein were up-regulated in the FS+baclofen group but were down-regulated in the FS+phaclofen group compared with those in the FS group. CONCLUSIONS: GABA_BR modulated the expression of H_2S/ CBS system in recurrent FS.

OBJECTIVE: To investigate the effects of recombinal human connective tissue growth factor(rhCTGF) stimulation on epithelial-myofibroblast transdifferentiation (EMT) and collagen-synthesis in human renal tubular epithelial cell line(HK2) in vitro. METHODS: The cultured HK2 cells were stimulated with rhCTGF of 5 ng/mL.The morphological changes were observed under an inverted microscope. The cells were collected at 0, 3, 6, 12, 24 and 48 hrs after rhCTGF stimulation. The expression of E-cadherin, α-smooth muscle actin (α-SMA), collagen Iα_1 (Col Iα_1) and collagen IVα_1(Col IVα_1) mRNAs were detected by RT-PCR. RESULTS: rhCTGF stimulation changed the HK2 cell appearance from oval to fusiform, down-regulated the E-cadherin mRNA expression and up-regulated α-SMA mRNA expression, but had no effects the Col Iα_1 and Col IVα_1 mRNA expression. CONCLUSIONS: Exogenous CTGF can mediate the EMT but has no collagen-synthesis effects on HK2 cells.

OBJECTIVE: Aquaporin (AQP) is a group of cell membrane transporting proteins. The study was designed to investigate the changes of AQP_1 and AQP_5 in the lung tissue under hyperoxia and their roles in pulmonary edema. METHODS: Two hundred newborn rats were randomized into different oxygen concentrations exposure: FiO_2=0.80 (Experimental group 1), FiO_2=0.60 (Experimental group 2), FiO_2=0.40 (Experimental group 3) and FiO_2=0.21 (Air control group). Rats were sacrificed at 1, 3, 5, 7 and 14 days after the beginning of experiment (10 rats each time point). The expressions of AQP_1 and AQP_5 were examined by Western Blot. The ratio of lung wet weight to lung dry weight (wet-to-dry weight ratio, W/D), and the protein content in bronchoalveolar lavage fluid (BALF) were measured. RESULTS: Compared with the Air control group, the W/D ratio and the protein content in BALF in the three experiment groups increased significantly and the increased extent was positively related to the duration and the oxygen concentration of hyperoxia-exposure. The expression of AQP_1 in the experimental groups began to decrease at the 3rd day and significant differences were found at the 5th and the 7th days after hyperoxia-exposure compared with that in the Air control group (P<0.05). The AQP_1 expression was restored somewhat at the 14th day after hyperoxia-exposure, but it was still lower in the Experimental groups 1 and 2 than that in the Air control group (P<0.05). The expression of AQP_5 in the experimental groups were reduced compared with that in the Air control group 3 days after hyperoxia-exposure and the decrease of AQP_5 expression was associated with duration of hyperoxia-exposure. The comparison among three experimental groups showed that the decrease of AQP_1 and AQP_5 expressions was associated with the concentration of hyperoxia-exposure. CONSUIONS: The expressions of AQP_1 and AQP_5 decreased in hyperoxia-induced lung injury and correlated with the severity of pulmonary edema.

OBJECTIVE: Glucocorticoid can induce apoptosis by regulating some genes' expression,but its effect on the pro-apoptotic protein Puma in leukemia remains unclear. This study was designed to investigate the effect of glucocorticoid on the expression of Puma in glucocorticoid-induced apoptosis of acute lymphoblastic leukemia cells in vitro. METHODS: Leukemia cells from acute lymphoblastic leukemia patients were transplanted into the immunodeficient mice. The transplanted leukemia cells were collected and then were treated with dexamethasone at the final concentration of 1 μM. The expression of Puma protein and mRNA in leukemia cells after dexamethasone treatment was detected by Western Blot and real-time quantitative PCR.A drug sensitive test was performed by MTT assay. RESULTS: The leukemia cells which came from the patients who had good clinical outcomes were sensitive to dexamethasone, while the ones from the patients who had poor clinical outcomes were resistant to dexamethasone in vitro. The sensitivity to dexamethasone in vitro between the resistant and sensitive leukemia cells was significantly different (P<0.05). No upregulation of Puma protein and mRNA was detected in resistant and sensitive leukemia cells after dexamethasone treatment. CONCLUSIONS: Glucocorticoid can not upregulate the expression of Puma in glucocorticoid-induced apoptosis of acute lymphoblastic leukemia cells in vitro.

No abstract available

No abstract available

No abstract available

No abstract available

No abstract available

No abstract available

No abstract available

No abstract available

No abstract available